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  1. Today
  2. What is CD molecule? CD molecule is the surface marker of white blood cells (including platelets and vascular endothelial cells, etc.) which appears or disappears in different lineages and stages of normal differentiation and maturation as well as during activation. Most of them are membrane-penetrating proteins or glycoproteins, including extracellular, membrane-penetrating and intracellular areas. A few CD molecules are carbohydrate hapten. Many CD molecules are functional molecules, some are enzymes, some are receptors, and some are signal transducers, ion channels or regulatory molecules. The role of CD Molecule CD molecule participates in some important physiological and pathological processes, such as: (1) mutual recognition of immune cells during immune response, recognition, activation, proliferation and differentiation of immune cell antigens, and exertion of immune function; (2) regulation of hematopoietic differentiation and hematopoietic process; (3) inflammation; and (4) migration of cells, such as metastasis of cancer cells. CD antigen and its corresponding monoclonal antibodies have been widely used in basic and clinical immunology research. In basic immunology, CD is mainly applied to: (1) gene cloning of CD antigen, discovery of new CD antigen and new ligand; (2) relationship between structure and function of CD antigen; (3) transmission of cell activation pathway and membrane signal; (4) regulation of cell differentiation; and (5) function of cell subsets. In clinical immunology research, CD monoclonal antibody can be used for: (1) detection of immune function of organism; (2) immunotyping of leukemia and lymphoma; (3) immunotoxin for cancer treatment, bone marrow transplantation and transplantation rejection prevention; and (4) immunomodulation therapy in vivo. Some important CD molecules CD Molecules Related to Antigen Presentation The CD molecule involved in antigen presentation mainly includes CD molecule constituting immunoglobulin Fc receptor and complement receptor and CD1. CD1 is the first human leukocyte surface differentiation antigen molecule detected by monoclonal antibody. CD1 is type I transmembrane glycoprotein. Its spatial structure is similar to that of MHC class I molecule and is linked to beta 2m, but its amino acid sequence is very different. According to the different amino acid sequences and glycosylation sites, CD1 molecules can be divided into CD1 (a, b, c) (category I) and CD1d (category II). CD1 molecule is mainly expressed on the surface of full-time APC, including most B cells, activated monocytes, Langerhans cells and dendritic cells. CD1 molecule is also expressed on immature double-negative thymocytes and CD8 T cells in the thymic cortex. CD1 molecule can present non-peptide antigens. The major CD molecules involved in T cell antigen recognition and activation include TCR-CD3 complex: TCR binds to CD3 molecules on cell surface to form TCR-CD3 complex. TCR recognizes specific antigen peptide-MHC molecular complex. CD3 molecules transmit signals and promote T cell activation. CD4: CD4 belongs to Ig superfamily and is a single-chain glycoprotein on the surface of cell membrane. Human CD4 molecule consists of 458 amino acid residues, including signal peptide 23 amino acid residues, extracellular domain 374 amino acid residues, transmembrane domain 21 amino acid residues, and intracytoplasmic domain 40 amino acid residues. There are four IgV-like functional areas in the extracellular area, including two glycosylation sites. CD4 is expressed on the surface of some T lymphocytes and thymocytes as well as some B lymphocytes, EBV-transformed B cells, mononuclear phagocytes and brain cells. The first and second functional regions of CD4 bind to the non-polymorphic parts of MHCII molecules, which can stabilize the interaction between MHCII-restricted T cells and APC cells with MHCII-antigen peptide complex. Therefore, CD4 molecules are also known as co-receptors involved in T cell activation. CD4 is also the main receptor of human immunodeficiency virus (HIV). The cytoplasmic region of CD4 molecule is linked to protein tyrosine kinase p56, which plays an important role in T cell signal transduction. CD8: CD8 molecule is a membrane-penetrating glycoprotein composed of a and beta polypeptide chains. The molecular weight of a chain is 34 kDa and that of beta chain is 30 kDa. Each chain consists of an IgV-like functional area, a ligand, a membrane-penetrating area and a cytoplasmic area. CD8 molecules are distributed in some T lymphocytes and thymocytes. The biological functions of CD8 molecule include: (1) as cell-cell adhesion molecule: MHCI class antigen is the ligand of CD8 molecule. CD8 binding with MHC class I can stabilize the binding of MHC class I restricted T cells (mainly CTL) to target cells with MHC class I molecule-antigen peptide complex. Therefore, CD8 is also known as a co-receptor involved in CTL cell activation. (2) After TCR binds to ligands, the intracellular domain of CD8 molecule is rapidly phosphorylated, leading to the activation of protein tyrosine kinase p56lck linked to CD8, which plays an important role in signal transduction of T cell proliferation and differentiation. Some drugs about CD molecule Clenoliximab Recombinant chimeric (primate/human) antibody expressed in CHO binding to human CD4/p55. Clenoliximab is a monoclonal antibody against CD4. It acts as an immunomodulator and has been investigated for the treatment of rheumatoid arthritis. The drug is a chimeric antibody from Macaca irus and Homo sapiens. Tregalizumab Recombinant monoclonal antibody to CD4. Tregalizumab is an immunomodulator. It binds to CD4. CD38 Daratumumab Recombinant monoclonal antibody to CD38. Daratumumab is an investigational anti-cancer drug. It binds to CD38. Keliximab Recombinant monoclonal antibody to CD4. Keliximab is a monoclonal antibody for the treatment of severe chronic asthma. It suppresses the immune reaction by binding to white blood cells via the protein CD4. The drug is a chimeric antibody from Macaca irus and Homo sapiens. Zanolimumab Recombinant monoclonal antibody to CD4. Zanolimumab (expected trade name HuMax-CD4) is a human monoclonal antibody and an immunosuppressive drug. It is being developed for the treatment of rheumatoid arthritis, psoriasis, melanoma, cutaneous and peripheral T-cell lymphoma. As the core component of hematopoietic immune tissue, human white blood cells have important biological functions. The study of differentiation antigens is not only helpful to understand the molecular structure, function, marker for cell subsets and differentiation stages, but also helpful to elucidate the differentiation, maturation, function and immunoregulation mechanism of immune cells. Therefore, it can deepen the understanding of immune response and immune diseases, and provide conditions for exploring new ways of clinical immunodiagnosis and treatment.
  3. Yesterday
  4. Bromelain is a pure natural plant protease obtained by extracting pineapple fruit stems, leaves and skins, purifying, purifying, concentrating, enzymatically immobilizing and freeze-drying. Its appearance is light gray powder, with a molecular weight of 33,000 and an isoelectric point of 9.55. The best quality bromelain is obtained by processing the middle stem of pineapple, filtering and concentrating by ultrafiltration method, and freezing and drying at low temperature. Products have been widely used in food, medicine and other industries. Application Application of bromelain in food processing industry 1. Baked food: Bromelain is added to the dough to degrade the gluten, and the dough is softened and easy to process. It can improve the taste and quality of biscuits and bread. 2. Cheese: used for the coagulation of casein. 3. Tenderization of meat: Bromelain hydrolyzes macromolecular proteins of meat proteins into easily absorbed small molecule amino acids and proteins. Can be widely used in the finishing of meat products. 4. Bromelain has been used in other food processing industries. Bromelain has been used to increase the PDI and NSI values of bean cake and soy flour to produce soluble protein products and breakfast, cereals and beverages containing soy flour. Others produce dehydrated beans, baby food and margarine; clarify apple juice; make soft candy; provide digestible food for patients; add flavor to everyday foods. Application of bromelain in medicine and health care products 1. Inhibit the growth of tumor cells According to relevant clinical research observations, bromelain can inhibit the growth of tumor cells. 2. Prevention and treatment of cardiovascular diseases Bromelain is beneficial as a proteolytic enzyme for the prevention and treatment of cardiovascular diseases. It inhibits heart attacks and strokes caused by platelet aggregation, relieves symptoms of angina, relieves arterial contractions, and accelerates the breakdown of fibrinogen. 3. For burn dislocation Bromelain selectively removes skin and allows new skin grafts to be performed as early as possible. Animal experiments have shown that bromelain has no adverse effects on adjacent normal skin. Topical use of antibiotics does not affect the effect of bromelain. 4. Anti-inflammatory effect Bromelain is effective in treating inflammation and edema (including thrombophlebitis, skeletal muscle damage, hematoma, stomatitis, diabetic ulcers, and sports injuries) in various tissues. Bromelain has the potential to activate inflammatory responses. Bromelain can also treat diarrhea. 5. Improve drug absorption Combining bromelain with various antibiotics (such as tetracycline, amoxicillin, etc.) can improve its efficacy. Related studies have shown that it can promote the transmission of antibiotics at the site of infection, thereby reducing the amount of antibiotics used. It is inferred that there is a similar effect on anticancer drugs. In addition, bromelain promotes the absorption of nutrients. Application of bromelain in beauty and cosmetics industry Bromelain has excellent effects on skin rejuvenation and whitening. Basic principle: Bromelain acts on the aged stratum corneum on human skin, causing it to degenerate, decomposition, removal, promote skin metabolism, reduce skin color darkness caused by sun exposure. Make skin care a good white and tender state. Application of bromelain preparation in feed Adding bromelain to the feed formulation or directly mixing it in the feed can greatly increase protein utilization and conversion, and can develop a wider protein source, thereby reducing feed costs. Action conditions: Optimum temperature: 53±1°C Deactivation temperature: ≥65°C The best PH value: 5.0 ~ 8.0 Inactivated pH: less than 3.0 and large with 9.5 Recommended dosage: 0.05 to 0.1% for protein meter Temperature range: 50 ~ 60 ° C Function PH value: 6.0~9.0 package: There are two types of packaging: 1) Aluminum foil bag packaging, net weight 1 kg / bag. 20 kg / carton. 2) Cardboard drum packaging, lined with transparent plastic bag, net weight 25 kg / barrel. Storage and storage period: It is recommended to store in a cool and dry place, the relative humidity is less than 60%, the temperature is lower than 25 °C; the storage period is 12 months, the storage period is 5 months at 5~15 °C in cool, low temperature and dry environment. It is best to keep away from odorous substances. About us Our products are used worldwide in academic, commercial, and government laboratories in diverse applications, including catalase, diamine oxidase, chitinase, and collagen. As a reliable supplier, Creative Enzymes supplies the products of high quality and competitive cost performance. We cooperate with a large number of satisfied customers in corresponding fields all over the world.
  5. Last week
  6. The main biological function of GC is to combine and transport vitamin D and its metabolites. Each GC molecule contains a VD enzyme binding site, and normally only 1% to 2% of the sites participate in VD metabolism. GC participates in the transport of VD and its metabolites between blood and cell membranes, so most of the plasma is free 6C. GC can bind to actin in vivo or in vitro to form a 1:1 complex, which greatly increases the molecular weight and increases the electrophoretic migration string. The isoelectric point of the Gc actin complex is lower than that of free GC. Gc binding to actin facilitates the clearance of actin from tissues. Vitamin D binding protein (also known as Gc protein), is an alpha globulin, a group of multi-gene super-including protein (ALB), prealbumin (PA), alpha-fetoprotein (AFP). The family, most of which is secreted by the liver parenchymal cells, has a relative molecular mass of about 55,000. It is abundant in serum and has many physiological functions. It not only binds vitamin D and clears actin, but also enhances C5 to neutral particles. The chemotactic activity of inflammatory cells such as cells, and activation of macrophages, regulation of osteoclast activity and transport of fatty acids and endotoxins, may also be involved in viral infection. [What is the function of vitamin D binding protein?] 1 Structure of vitamin D receptor VDR is a member of the nuclear receptor superfamily such as thyroxine and corticosteroids. VDR has a large similarity in sequence and structure to subfamilies including retinal, thyroid hormone, and peroxisome proliferator-activated receptors. The study confirmed that the human VDR gene is located on chromosome 12, consisting of 11 exons and several introns, about 75 kb in length, and the protein consists of 427 amino acids. From the amino end to the carboxy end, it can be generally divided into six functional regions: A, B, C, D, E, and F. The AB region is a ligand-independent, tissue-specific transcriptional activation self-regulating functional region AF1. The human VDR AB region consists of approximately 24 amino acid residues. No features were found in this domain. Region C: DNA binding domain DBD, involved in DNA sequence recognition, and is also partially involved in the formation of dimer interfaces. This region is highly conserved and the homology of human, rat and chicken VDR DBD is 98.15%. Zone E: This region is relatively large and functional: participating in binding ligands, thus referred to as the ligand binding domain LBD; forming a dimer with RXR; forming a ligand-dependent transcriptional activation or inhibition functional region AF2. In addition, the E region also has a synergistic effect on DNA recognition. Zone D and Zone F: The functions of these two zones are unknown. Zone D may be a hinge region that primarily regulates the flexibility of the receptor and may be related to nuclear localization. 2 VDR regulates the calcium-binding protein gene VDR regulates many genes, most of which are genes in bone metabolism. Here we use the calcium-binding protein gene as an example to introduce the regulation of VDR genes. Calcium is second only to oxygen, carbon, hydrogen and nitrogen in the body, accounting for about 2% of the human body. Calcium absorption is a complex physiological and biochemical process, including active transport and passive diffusion. Active transport is the main factor when the body consumes less calcium. Active transport of calcium requires the assistance of vitamin D and calcium binding proteins. Wasserman et al. first confirmed the formation of specific calcium-binding protein (CaBP) after treatment of rickets with vitamin D. This vitamin D-induced CaBP has been found in more than a dozen animals and is species-specific. The molecular weight of human intestinal calcium CaBP is 12000-21000, which has a high affinity with calcium, and the ability to bind calcium is about 2×10-5M-1. CaBP is highly concentrated on the absorption cells of the goblet cells and the brush border of the small intestine. In different parts of the intestine, intestinal CaBP level and calcium absorption rate are consistent, with the highest in the duodenum, the second in the jejunum, and the lowest in the ileum. In addition, CaBP is also present in tissues such as the kidney, hypothalamus, cerebral cortex, adrenal gland, and parathyroid glands. CaBP plays an important role in the process of vitamin D-mediated intestinal calcium transport. Mammals can synthesize two CaBPs, namely CaBPD28K present in the kidney and central nervous system and CaBPD9K in the small intestine. Injection of 125OH2D3 into vitamin D-deficient rats and chicks resulted in rapid synthesis of CaBP mRNA and accumulation of CaBPD9K mRNA in intestinal mucosa and kidney cells. About us We provide a wide range of high quality normal human and animal cells, cell culture medium and reagents, FISH probes, tissue arrays, microorganisms and equipment. In addition, we also offer series of related services including cell services, biosample services and histology services for the researcher to make their project better and faster. Here are some products: FSTL5, FTCD, FTL, FTO, etc.
  7. Chronic prostatitis includes chronic bacterial prostatitis and non-bacterial prostatitis. Among them, chronic bacterial prostatitis is mainly pathogen infection, mainly retrograde infection, the pathogen is mainly Staphylococcus, often repeated urinary tract infection attack history or prostate massage fluid in the persistent presence of pathogenic bacteria. Non-bacterial prostatitis is a complicated pathological change caused by many complicated causes and inducements, including inflammation, immunity and neuroendocrine. The main clinical manifestation is urethral irritation and chronic pelvic pain. And often associated with psychological symptoms of the disease, clinical manifestations are diverse. And the name of chronic prostatitis belongs to the old classfication system in which prostatitis is divided into acute bacterial prostatitis, chronic nonbacterial prostatiti and prostatodynia. How much do you know chronic bacterial prostatitis? The pathogenic factor of chronic bacterial prostatitis is mainly due to pathogen infection. When the body resistance is stronger or the pathogen virulence is weak, retrograde infection is the main form of infection. The main pathogens were Staphylococcus, followed by Escherichia coli, Corynebacterium and Enterococcus. Prostatic calculi and urinary reflux may be important causes of persistent pathogens and recurrence of infection. How much do you know chronic non-bacterial prostatitis? The nosogenesis of chronic non-bacterial prostatitis is still unkown. The nosogenesis of chronic non-bacterial prostatitis is very complicated with widespread controversy. Many hold a view that the reason that it may be caused by an initiator or may be multifactorial in the first place or more of which play a key role and interact with each other. Also, it may be many different diseases that are difficult to distinguish but with identical or similar clinical manifestations. What’s more, these diseases have been cured and the damage it causes continues to work independently with pathological changes. Most scholars believe that the main cause of the disease may be pathogen infection, inflammation and abnormal pelvic floor neuromuscular activity and immune abnormalities. How manyoncogenesis do you know?pathogen infection Although the pathogen can not be isolated by routine bacterial examination, it may still be related to some special pathogens, such as anaerobes, L-Proteus, nanobacteria, chlamydia trachomatis, mycoplasma and so on. Some studies have shown that the detection rate of local prokaryote DNA in this type of patients can be as high as 77%, and some clinical cases of "aseptic" prostatitis, which are mainly chronic inflammation and repeated attacks or aggravation, may be related to these pathogens. Other pathogens, such as parasites, fungi, viruses, trichomonas and Mycobacterium tuberculosis, may also be important pathogenic factors, but there is no reliable evidence, so far there is no unified opinion. Urinary dysfunction Some factors cause excessive contraction of the urethral sphincter, resulting in bladder outlet obstruction and residual urine formation, resulting in urine reverse flow into the prostate. Not only can the pathogen go into the prostate, but also directly stimulate the prostate to induce aseptic "chemical prostatitis", causing abnormal urination and pelvic pain. Many prostatitis patients have a variety of urodynamic changes, such as decreased urinary flow rate, functional urinary tract obstruction, detrusor-urethral sphincter synergy, and so on. These abnormalities may only be a clinical phenomenon, and their nature may be related to underlying pathogenic factors. Psychological factors More than half of the patients with chronic prostatitis have obvious psycho-psychological factors and personality changes, for example, anxiety, depression, hypochondria, hysteria, even suicidal tendency. The changes of these mental and psychological factors can cause autonomic nerve dysfunction to cause posterior urethral neuromuscular dysfunction, and lead to pelvic area pain and urination dysfunction or hypothalamus-pituitary-gonadal axis changes and affect sexual function. The further aggravate symptoms eliminate mental tension can make symptoms relief or recovery. However, it is not clear whether psycho-psychological change is its direct cause or secondary manifestation. Neuroendocrine factors Patients with prostate pain are prone to fluctuations in heart rate and blood pressure, suggesting that autonomic nervous responses may be involved. The pain is characterized by visceral organ pain, local pathological stimulation of the prostate and urethra, triggering of spinal cord reflex through the afferent nerve of the prostate, activation of astrocytes in the lumbar and sacral spinal cord. Nerve impulses pass through the reproductive femoral nerve and ilioinguinal nerve efferent impulses. The sympathetic nerve endings release norepinephrine, prostaglandin, calcitonin gene-related peptide, substance P and so on, which cause bladder urethral dysfunction and lead to perineum. Abnormal pelvic floor muscle activity, persistent pain and traction in the corresponding area outside the prostate A wrench. Immune response abnormality Recent studies have shown that immune factors play a very important role in the development and progression of III prostatitis. Some cytokines may change in prostatic fluid, seminal plasma, tissue fluid or blood of the patient, such as IL- 12 Family,IL- 7 Family, IL-8 Family, IL-10 Family, TNF- α and MCP-1. The level of IL-10 was positively correlated with the pain symptoms of patients with III prostatitis, and immunosuppressive therapy was effective. Oxidative stress theory Under normal conditions, the production, utilization and removal of oxygen free radicals are in dynamic equilibrium. The excessive production of oxygen free radicals and the decrease of scavenging effect of free radicals in patients with prostatitis may be one of the pathogenetic mechanisms, which can decrease the response ability of the body to antioxidant stress and increase the production and by-products of oxidative stress. Pelvic disease factors Some patients with prostatitis are often associated with prostatic peripheral venous plexus dilatation, hemorrhoids, varicocele and so on, suggesting that the symptoms of some patients with chronic prostatitis may be associated with pelvic venous congestion and blood stagnation. This may also be one of the reasons for the persistence of governance.
  8. How to Get the Most Out of Solar Panels

    If you have recently decided to install solar panels, then you need to start thinking about some ways that you can make the most out of that technology. Even though modern solar panels require very little maintenance, quite a bit can be done to improve their efficiency and prevent unnecessary damage. Here is a look at a few tips and tricks that will take your solar energy system to the next level. Feed Less Energy Back Into the Grid Using the energy as it is being produced is going to reduce waste and lower your overall bills. That is why many experts suggest using high-consumption devices during the middle of the day. That includes your dishwasher, washing machine, and dryer. You should also consider upgrading to an electric water heater to further improve the efficiency of your home and reduce your monthly expenses. Install a Backup Battery In addition to being more affordable than ever, residential lithium batteries are also exceedingly efficient. With a high-quality battery system, you might be able to use almost 100 percent of the energy that your solar panels produce. If your local power company increases the cost of electricity during certain hours of the day, then you can simply switch over to the battery during those hours and save some money. Installing a backup battery is a relatively simple project, and an experienced contractor should be able to finish the job in a single day. Maintain the Panels Themselves As a general rule, you should carefully inspect all of your solar panels at least two or three times a week. You want to be absolutely sure that the panels aren’t caked in mud, covered in leaves, or under thick branches. Depending on where you live, you might need to clean the panels once or twice a month as well. Most dirty panels can easily be cleaned with a hose, but you might need to use a sponge and some warm water with a little bit of soap in it if there is quite a bit of grime. Upgrade Your Roof Before your panels are installed, you should consider contacting a company such as Landmark Roofing to upgrade your roof. A roofing company can replace your old tiles with metal sheets and improve the angle of the roof itself. In most locations, solar panels work best when the roof is between 30 and 45 degrees. Panels that are perfectly flat will be exposed to sunlight for longer stretches of time, but the energy production is going to be much lower. Once your panels have been installed, you need to keep a close eye on your energy consumption each and every day. Sudden changes to your input or output could be the result of a serious issue that needs to be addressed.
  9. Bispecific antibody is known as bsAb for short. The branch of technology in antibody drugs and bispecific antibody drugs over the past few years have been absolutely the hot spot. Anyone who makes drugs feels like it's hard to say not knowing this technology. Pharmaceutical companies feel like they might be inferior if they don't have a layout in this direction, and they're likely to miss out on big opportunities in the future. Is that the technology really important? Since the technical analysis is very professional, let me talk about it from the market. The two bispecific antibodies which is currently available are Removab and Blincyto. Let's take a look at two stories about them. Do you know First-in-Class is with complicated background and awkward status? Removab, the first therapeutic antibody drug in the field of bispecific antibodies who was approved by the European Union on April 20, 2009, is certain to be First-in-Class consisting by antibody-peptide. Its original patent was granted in 1998 by Ascension GmbH, a smaller German biotechnology company. Shortly, after the deal was completed, TRION entered into an alliance with Fessenius, an established German company, and decided to jointly develop the later approved listing Removab. Ascension GmbH later sold all of its TRION holdings to Fessenius, a biotech subsidiary focused on developing biopharmaceuticals, which was later sold to Fesenius by Ascension GmbH, a subsidiary that specialized in the development of biological drugs. In 2011, Swedish Orphan Biovitum (Sobi) struck a marketing deal with Fresenius Biotech on which Removab was successfully approved for listing. Removab was approved for sale by EMA in June 2013. Also, in this month, Fessenius announced the sale of Fresenius Biotech would become as a whole to the Further family, the owner of Neopharm (the amount of the deal was not disclosed by the parties to the agreement). Fresenius Biotech GmbH was logically renamed Neovii Biotech GmbH. The indication for Removab approved in 2009 in the European Union is malignant ascites. If there is also a therapeutic effect on primary tumors, the market is worth imagining. But the market reaction was completely different. In 2009, the product was sold at $1.66 million, and in 2010-2012 it was 3.32 / 4.43 / 4.54 million dollars, respectively. The annual sales of the product have not been found in mainstream medical databases since 2013, followed by ovarian cancer, stomach cancer, non-small cell lung cancer, breast cancer, and other indications. In the II phase, dismal sales figures offer no hope for these new indications. But Removab, has never been born. Its biological parents, Ascension GmnH, had no attachment to it from the start. Its two stepfathers, TRION and Fresenius, were just passing by in the process of his growth. And Neovii turned a blind eye to him in the end. How is the market condition of Blincyto? Let's take a look at the Blincyto of Amgen approved by FDA and EMA in December 2014, which is a double-specific antibody to CD3 on one end of CD19 that binds to the surface of white blood cell B cells, and on the other end of the surface of T cells. The basic idea is consistent with that of another popular cellular immunotherapy now. Its first indication is the second-line treatment of acute lymphoblastic leukemia. Blincyto has another American biology company, Micromet Inc. At the time, it was named MT103. In June 2003, Micromet signed an agreement with MedImmune to develop MT103. As everyone knows, in March 2009, after AstraZeneca acquired MedImmune2 in June 2007, Micromet bought back an interest in the original MedImmune agreement from AstraZeneca, so Micromet regained ownership. In January 2012, Amgen became Amgen's AMG103 by buying Micromet’s MT103 in full cash for $11 a share. Later, in May 2013, Amgen licensed the Japanese interest in the product to Astellas. In August 2013, it licensed its Indian interest to Dr. Reddy's in India. Blincyto's U.S. sales were about $50 million in 2015, compared with $85 million in 2016. At the same time, Amgen is also carrying out clinical research on new indications such as diffuse large B-cell lymphoma and mantle cell lymphoma, which is expected to be further expanded. Some analysts believe the drug could reach annual sales of $178 million in 2017. Amgen spent $1.16 billion in 2012 and certainly there would not spend small amount money on research and drug developing after the acquisition. Amgen's global sales figures for 2016 were $225 billion to $228 billion. However, Amgen will no doubt be a bit disappointed by the result of Blincyto alone in 2016. Why the prospect of a variety on the market is not bright? As for the next bispecific antibody drug most likely show in the market, Roche's ACE-910 (RG6013 Emicizumab) is considered to have no bright future. The drug is originally developed by Japanese and foreign pharmaceutical companies and they reached a global cooperative development agreement with Roche in August 2014. Although severe adverse events with thromboembolism / thrombotic microvascular disease (Thrombotic microangiopathy, TMA) were reported in four participants in November 2016, the possibility that ACE-910 would eventually be approved for listing is high. The indication for ACE910 is type A hemophilia with VIII inhibitors. We all know that hemophilia is a rare disease and the global market for hemophilia is estimated to be around $10 billion. But this huge market has been crowded into a number of heavyweights, including Shire,Baxalta, Northrop, Bayer, Baijian and CSL, of which Type A hemophilic drugs (such as Feiba,Advate,Adynovate) sold $2.84 billion in 2015. We can see that this market is large. ACE-910 expects to hit the market around the end of 2019, which means we don't see a double antibody drug with annual sales of more than $500m over the next five years. Bispecifics, how many people are obsessed with you? How many people are manic for you? However, to me, it is really hard to say I love you.
  10. Earlier
  11. Abstract: In the long process of antibody research, three generations of different levels of antibody preparation techniques have been developed. A polyclonal antibody prepared by antigen-immunizing higher vertebrates is called a first-generation antibody; a monoclonal antibody produced by hybridoma technology only for a specific antigenic determinant is called a second-generation antibody; and recombinant DNA is applied; The technique or the method of gene mutation modifies the coding sequence of an antibody gene to produce an antibody protein molecule originally existing in nature called a genetically engineered antibody, that is, a third generation antibody. This genetic engineering is called antibody engineering. Human hybridoma cells have been tried in the early days to produce human monoclonal antibodies, but this technique has rarely been used due to the instability of human hybridoma cells, the low affinity of human monoclonal antibodies, and ethical controversy. In the mid-1980s, new technologies were introduced to combine the genetic structure and function of immunoglobulins with DNA recombination technology, and then the recombinant immunoglobulin genes were introduced into cells for expression. In the third generation of antibodies, mainly including humanized antibodies, small molecule antibodies, antibody fusion proteins and some specific types of antibodies, to some extent overcome the shortcomings of the first two generations of antibody technology. In addition, the construction of a phage antibody library, a ribosome display library, and the like allows specific antibodies to be obtained without antigen immunization. Recombinant antibody technology makes it possible to prepare humanized antibodies and human antibodies, which is not possible with other conventional polyclonal or monoclonal antibody preparation methods. Keywords: recombinant antibody, monoclonal antibody, phage antibody technology Chimeric antibody One method of reducing the immunogenicity of a murine monoclonal antibody is to link the variable region of the murine immunoglobulin to the constant region of the corresponding human immunoglobulin, thus producing a murine-human chimeric antibody, human region in 60%~70%. Using the recombinant DNA technology, the variable region gene of the murine monoclonal antibody is ligated to the human constant region gene, and the constructed chimeric gene is inserted into an appropriate expression plasmid, and then transfected into the corresponding cells for expression. The chimeric antibody produced has the function of binding antigen, and at the same time reduces the heterogeneity of the murine monoclonal antibody, such chimeric antibody is not much different in affinity from the corresponding murine monoclonal antibody, and humans have it The immune response will be reduced. However, since it still retains the heterogeneity of the murine immunoglobulin variable region, clinical experiments have shown that the chimeric antibody also produces an immunological response to the variable region of human anti-chimeric antibody (HACA) when applied. Small molecule antibody In order to break through the limitations imposed by the oversize of monoclonal antibodies, techniques for the development of small molecule antibodies have been developed. The goal of these techniques is to obtain antigen-binding fragments (Fabs) of antibodies and variable regions (Fv) of antibodies. Such a fragment can be obtained by cleavage of an antibody, or by amplifying a gene associated with an immunoglobulin and cloning and expressing it in bacteria. Small molecule antibodies have small molecular weight, strong penetrability, low immunogenicity, and short half-life. At present, there are several studies and more practical prospects. Single chain antibody The technology links a heavy chain variable region and a light chain variable region with a ligation peptide, and is expressed as a single-stranded polypeptide by a prokaryotic expression system and folded into a novel antibody consisting of a heavy chain and a light chain variable region. Multivalent antibody The multiple antigen binding sites of the antibody have different specificities and are capable of binding different antigen molecules, which changes the deficiency that the traditional antibody can only bind to a single antigen molecule. At present, researchers are paying more attention to the application of bispecific monoclonal antibodies. Fab fragment In the antibody, the Fab segment is mainly used to combine the role of the antigen. Phage antibody technology In the early 1990s, based on the PCR technology, the expression of antibody Fab fragments in E. coli and the rapid development of phage display technology, antibody library technology based on molecular biological methods appeared. Among them, the phage antibody library technology is superior. The basic procedure of phage antibody technology is to amplify a full set of antibody variable region genes or antibody fragments (Fab, Fv or seFv) genes by PCR and transfer them to the vector after phage capsid protein gene, which can be filamentous. The coat protein of the phage forms a fusion protein, and the antibody Fab fragment or single-chain antibody is expressed on the surface of the phage, and then the specific antibody and the gene encoding the same are obtained from a plurality of antibodies by affinity screening of the antigen. A phage population cloned and assembled using a full complement of antibody variable region genes of B lymphocytes is referred to as a phage antibody library. The technology is simple to operate, does not require cell hybridization or complex PCR technology, has a short cycle and low cost; can simultaneously screen several antibodies using different antigens, and can be selected in millions to hundreds of millions of molecules; suitable for antibodies, hormones The preparation of proteins such as enzymes, drugs, and random polypeptides is more interesting because the technology can obtain antibodies that cannot be produced by immune animals due to the influence of immune tolerance mechanisms. Phage antibody technology has greatly promoted the development of genetically engineered antibodies, and their production has brought new hopes for the prevention, diagnosis and treatment of human and animal diseases. References [1] Schillberg S, Fischer R, Emans N. Molecular farming of recombinant antibodies in plants [J]. Cell Mol Life Sci, 2003, 60(3): 433-445 [2]Bouquin T, Thomsen M, Nielsen L K, et al. Human anti-rhesus D IgGI antibody probuced in transgenic plants[J]. Transgenic Res, 2002, 11(2):15-22
  12. A heat pump works well if you live in a moderate climate. This is because it’s more efficient than other forms of heating and cooling. Here are some of the benefits that a heat pump can offer when it comes to saving money by conserving energy. Temperature Stability Setting your heat pump for a consistent temperature is the most efficient use of this type of system. This is because of the fact that it can run for a minimal amount of time and maintain the temperature of your home. With a standard HVAC system, there tend to be fluctuations that require more energy when the system kicks on and off. A heat pump has an electric motor that requires very little energy in order to reap a stable temperature for your whole home. Heat Conversion The action of a heat pump is that is doesn’t generate heating or cooling within the unit itself. It simply converts the heat that exists in the air and transfers it into your home by mechanical means. This method allows it to be the most efficient possible with the least amount of energy usage. The benefit of heat pump installation in your home is that you get instant heat for very little drain on your electric bill. Cooling Potential Another benefit of having a heat pump is that it can also cool your home. This is simply the heat pump removing the built-up heat from inside your house and transferring it to the outdoors. The amount of energy that’s required for this function is less than your standard air conditioning unit would require. This allows for you to have a cool home without having to worry about a spike in your electric bill during the summer months. Reduced Electric Consumption The amount of electricity that’s utilized with your heat pump will depend on your climate and the model that you select. There are models that will use significantly less energy than others. For the vast majority, the energy usage is less than you would observe with a standard HVAC system. This allows you to save on your electric bill and work to conserve energy. On average, a heat pump uses a quarter of the amount of energy as the more traditional HVAC units. There are lots of benefits that you can reap from having a heat pump in your home. The energy savings are just the beginning when it comes to the ways that it can improve the comfort and sustainability of your household.
  13. Poor outdoor air quality is a serious concern because it can have both immediate and long-term effects on your health. It is easy to assume that outdoor air quality would not affect you while indoors, but this is not the case. Outdoor pollutants may enter the home through open doors and windows, smaller or larger gaps in the exterior of the home and more. These pollutants can combine with indoor pollutants to create truly unhealthy air quality in your home. By following these essential tips, you can promote healthier indoor air quality at home. Service Your HVAC System Your air conditioning and heating system is designed to circulate air throughout the home. By doing so, it also circulates pollutants. Over time, the components may get filthy, and this can contribute to poor indoor air quality. It may also cause energy inefficiency until you schedule heating and air conditioning repair service. In addition to keeping your HVAC system well-maintained, you should replace the air filters regularly for superior air quality. Use an Air Purifier Purifying your air essentially means that you are removing pollutants that ultimately may cause illness or disease. By investing in a high-quality air purifier, you can remove many potentially hazardous elements from your home’s air. Keep in mind that plants are natural air purifiers. In addition to having a purifying device, keeping at least a few houseplants in your home can have positive effects. Keep the Windows and Doors Closed While trace pollutants may enter your home through small gaps and fissures throughout the exterior, open doors and windows enable pollutants to stream into the home. You may think that opening doors and windows allows fresh air to enter the home, but the opposite may occur. Keeping doors and windows closed as much as possible is most beneficial for indoor air quality in many cases. Replacing seals around exterior doors and windows is also beneficial. Because you spend many long hours at home each day, you understandably need this air to be as clean and fresh as possible. Both indoor and outdoor pollutants can be problematic, and their level can increase over time. To improve indoor air quality, it is necessary to prevent outdoor pollutants from entering the home as much as well. It is equally essential to remove pollutants through various purification methods. By taking these steps, you can promote a healthier indoor environment and potentially keep many related health issues at bay.
  14. While buying a good quality pair of shoes may seem like something pretty obvious, it's not as easy as it sounds when you actually get into the store. Sure, style and looks have something to do with the right shoes, but there are a lot of other factors that need your consideration when you want to take care of your tootsies. Before you make any important shoe decisions, take a moment to think about your feet. The comfort of your feet has a tremendous impact on your overall comfort. Just think about it - sore feet, cold feet, hot feet, itchy feet - all of these things are enough to drive you to distraction. If your feet are uncomfortable, the rest of you is uncomfortable. Keep this in mind the next time you walk into a shoe store and look at those brand-name, yet uncomfortable shoes. You need to treat your feet right. This means that you not only have to buy a pair of shoes that look good, but you also need to find ones that fit your feet. This may sound obvious, but the majority of people wear shoes every day that aren't quite suited to their unique foot shape and size. So how do you find a pair of shoes that are suited to your unique foot shape and size? It's not as easy as it sounds. Just follow these steps: 1. Have the salesperson measure your feet. He or she should measure both the length and width of each of your feet. Your feet will likely be slightly different in size, and it is important to know this when you're seeking the ideal comfortable shoe. 2. Use the larger foot's size. While you can use an insole to make a shoe that is a bit too big fit your smaller foot, you'll only hurt your larger foot by stuffing it into a shoe that is a bit too small. 3. Make the shoe fit your toes and your heel. There should be enough room to wiggle, but not enough to slip around. 4. Walk about the store to be sure that there isn't any pinching or slipping. 5. Don't assume that the shoe will break in. Have them fit when you buy them. When it all comes down to it, shoes are your foot's best friend. Don't let your tootsies down! Article Source: https://www.import-express.com/apa/shoes.html
  15. When you want to cut your heating and cooling bills, selecting high-efficiency HVAC equipment makes a big difference. The U.S. Department of Energy (DOE) sets efficiency standards for HVAC systems because they’re the most energy-hungry of all home appliances. Better efficiency translates to lower demand for electricity and fuel and reduced energy bills for you. HVAC Efficiency Ratings When choosing central HVAC systems, look for high ratings. The DOE requires efficiency ratings on the equipment itself and its packaging. Higher is always better regardless of whether you’re buying a furnace or an air conditioner. The Energy Star and Most Efficient labels indicate products that use the least amount of energy for the best results. For example, a high-efficiency furnace uses measurably less fuel each time and requires less heating services because it runs better than regular models. Lower fuel usage helps preserve natural resources and saves you money. Making Energy Energy production exacts a toll on the planet, especially when the fuels to generate electricity include fossil fuels and nuclear power. While renewable resources like solar, wind and hydro have environmental impacts up front, the resources they use to produce electricity are clean. Fossil fuels, on the other hand, contribute to atmospheric warming on a continuous basis because of their byproducts. Nuclear is a clean source of energy but the nuclear waste it produces is one of the most dangerous substances on the planet. Power Production Efficiency If energy production were 100 percent efficient, using less efficient HVAC systems wouldn’t have the impact it does. However, power plants are not very efficient themselves. For example, coal-fired plants waste over 60 percent of the coal they consume. Nuclear and natural gas are slightly more efficient, while electricity produced by hydroelectric plants is nearly 100 percent efficient. The phrase experts use to describe this power plant efficiency is the heat rate. It describes how much of the fuel that’s used goes toward producing electricity as opposed to being wasted. The efficiency of furnaces and air conditioners follows the same principle. The most efficient systems waste less energy. Atmospheric Warming If the fuels used to produce electricity didn’t have waste products like carbon dioxide and nitrous oxide that warm the atmosphere, it wouldn’t be so important to use high-efficiency HVAC equipment. But when equipment needs more fuel and energy, the volume of gases going into the atmosphere increases. Since the heat they hold warms the planet, using more efficient HVAC systems helps reduce harmful greenhouse gases. Until all power plants are 100 percent efficient or use clean, renewable fuels, it will be important to conserve energy usage. HVAC systems are front and center for conservation efforts since they use the most energy of all home appliances. More efficient systems help you save energy without sacrificing your comfort.
  16. Do you have a deck outdoor and you want to decorate it for the upcoming festivals? Do you have any idea on how you can decorate your patio or deck? Are you capable of designing and decorating the deck on your own or you needing the help from professional decorator? Do you have decided any theme for the decoration? If not, here are some of the tips that you can follow to decorate your decks for the festivals. It is not always possible to build a new deck whenever the festival arrives; you can revamp or decorate it when required. Tips to Start Decorating Your Deck for the Upcoming Occasion- 1. Check out The Stock Before you start to revamp the space, you can check out what the items you will like to keep are and what items you have to discard. You must keep in mind that reusing the old items can help you to save money if you have a tight budget. Once, you have decided what items you require to keep and what items you have to discard, you can easily make a list. If you have old broken pots or the worn out seat cushions, you should throw them first. 2. Choose a Particular Theme The theme of the deck sets the tone for your deck. The theme can be the particular motif, items that you require for designing and the colour combination with which you have to design the deck. Not sure from where you can begin? Just check around you and choose the colour of the flowers in your garden or you can get inspiration from your favourite beach or the nearby park. 3. Clean and Restore the Deck Before starting to decorate your deck, you have to clean and restore a deck. You have to clean the wood by removing the debris and mold growth that may be developing during the winter months. You have to prep the deck by sealing. Clean the deck each time before decoration. Once you have cleaned the deck, you must go through the finish. You can perform the splash test, for example, you must splash the water on a few boards and watch whether the water gets absorbed quickly or not. If the water gets rapidly absorbed, this means that the wood is protected in the right manner and it is the high time to seal. 4. Decorate and Accessorise You can decorate the living space by bringing the indoor couches outside! You can use the décor items for creating a beautiful space for your family and guests. You can use the accent pillows and seat cushions that are ideal for providing utmost comfort. 5. Bring Potted Plants or Vertical Gardens You can also decorate your outdoor patio or deck with the help of the potted plants or the vertical gardens. They will bring a sense of greenery to space. You can also consult an interior decorator or designer to design or decorate your deck. These are some of the tips to follow if you want to decorate your deck. Revamp your patio or deck and create unforgettable memories with your friends.
  17. Ever wonder why dropship wholesalers for shoes are in much demand in today's market? The road to success for dropshipping shoes only takes sheer determination. Shoes are more than fashion items, it's a necessity. Who would want to go out to a supermarket or a club barefooted? With online selling at its peak, your chances of successfully selling shoes on the Internet is indeed high. However, finding your way to a trusted supplier can be an intimidating process. Here are insider secrets on how you can get your way to trusted dropship wholesalers. Visit a shoe manufacturer now. If you are lucky enough to be living in the same location of a shoe manufacturing company, you can get first hand information by visiting their office. You may not immediately get a deal, but you can get useful details, such as a list of the manufacturer's distributors and dropship wholesalers. This way you'll be able to get reliable suppliers for your online shoe store. Visit offices of dropship wholesalers for shoes. Dropship wholesalers do not usually have websites and this is a fact most first time sellers do not know. So, once you get hold of wholesaler info, contact or visit their office immediately. Your competitor might be researching the same shoe wholesaler, so you must find way to that wholesaler your first. Bring along with you relevant business documents and tax identification. Ask for samples, if available, but if not, you can always purchase a few pair of shoes as samples. While you have presented yourself as a legitimate seller, you also have the right to check the authenticity of the wholesale business by asking for business registration. Contact dropship wholesale list providers. The world of dropshipping is a secret world. That's why sellers themselves do not usually disclose information about the wholesalers they deal with. But there are those who are in the business of selling wholesaler lists. These people are also dropship sellers themselves. For a fixed amount, you can get access to hundreds of dropship wholesalers for shoes that offer competitive prices, including those coming from China. Trace the wholesaler by buying a product. You can get a little sneaky by buying a pair shoes from an online seller and inspecting the product details, which will more likely include the wholesaler info. With the wholesaler's name at hand, it will be easier for you to research the contact details of that supplier. From visiting a shoe manufacturer to tracing the product details, all it takes for you to find trusted wholesalers for shoes is common sense and a few tricks. Remember, your goal is not get to the wholesaler fast. It is about getting a reliable dropshipper. So, if one tip doesn't work, you still have 3 other options towards getting reliable dropship wholesalers for shoes. Article Source: https://www.import-express.com/apa/shoes.html
  18. Filtering jug or mineral water

    Every day we make a number of purchasing decisions, guided by many factors. What determines the choice of products that we bring from a store? Usually it is the same set of criteria: price, quality, brand awareness, habit and taste preferences. However, more and more often we are also trying to select products that are healthier and more ecological, because we are more and more aware of their impact on our health and life. Considering these criteria, will giving up mineral water for filtered water is a good choice? Purchase of a jug or a filter bottle is the cost of several dozen zlotys on average, and a filter for several dolars a month. Many calculations indicate that the purchase of a filtering jug can save several hundred dolars a year, taking into account the regular monthly filter change of course. In addition, we reduce the amount of rubbish, throwing up to 26 kg of plastic a year less. In this way we become more environmentally friendly, because, unfortunately, we still do not always have the option of waste segregation and guarantee of its proper recycling. According to German scientists from the health perspective water stored in PET packaging can be contaminated with substances that may cause for problems with the endocrine system (e.g. bisphenol A, which is so avoided by manufacturers of accessories for children). Products for filtering water at home available on the market are largely recyclable, manufactured with care for the environment and will serve us for many years. Thanks to the online purchase options, filtering jugs, bottles or filters will reach any place, even if these products may not be accessible easily. Online shopping also gives us access to the full range of products and provides detailed information, making it easier to choose the most suitable option for our household. Companies producing them have been known for years, so there is no fear that you will not be able to choose a filter or replace the filtering jug if necessary in the future We can always filter the water by simply filling our jug or tap water bottle without having to go out to the store. The quality of water bought in stores is a separate topic. To be sure what kind of water we are dealing with, we need to read the sum of minerals dissolved in it from the label. Mineral water is considered the one containing 1000 mg of mineral substances in 1 litter at least. Out of over 200 water brands available, only about 30 are mineral waters, the other ones are deep-sea or tap water, enriched with minerals. Thanks to the tap water filtration, we are sure that we consume purified, safe and tasty water, which is especially important if there are children at home. This is also an economical solution that supports an environmentally friendly approach to the household. Read more about water filtering systems on Dafi.us
  19. A jug with a filter or a bottle

    Currently, various solutions are available. Their choice depends on our needs. Filtering jugs are the most widely known and comes to our mind first, when we think about filtering water. The first devices of this type appeared in our homes many years ago. The history of filtering water at homes dates back to the 1970s when jugs with removable filters were the first solutions available in general. Jugs available at present vary not only in terms of design and colour, but also in their production process. Materials they are made from is the basic difference - jugs are made from glass and plastic. Each solution has its own advantages, so that you can choose the most suitable product for your household. You can also choose different filters for your jug, there are more universal filters as well as ones with a more specialized usage, e.g. for preparing cold drinks (the ones that better remove chlorine and organic impurities, easily perceptible in non-heat treated beverages) or for household appliances (with increased water hardness reduction). Let's choose the filter that best suits our needs. Keep in mind that all filters provide a high degree of water purification by removing substances disrupting the smell or taste thanks to activated charcoal filter and an ion exchange resin which removes heavy metals from the water and reduces water hardness. What else is available on the market? The bottle is ideal not only for active people, practising sports and requiring additional hydration during increased physical activity, but also for anyone who wants to live a healthy life. Light, ergonomic, it can be filled with tap water wherever we are, ensuring that the water consumed is clean and safe, free from harmful substances, sediments and mechanical impurities. Filters in bottles, like in jugs, are replaceable, thanks to which water quality will always be high. Additionally, one filter can replace the purchase of up to 300 bottles of water with a capacity of approx. 500 ml each, helping us to care both for the environment and the home budget. A filter bottle is a perfect complement to home filter systems, providing clean, filtered water whenever you do not have the access to a jug or installation with a flow filter. Due to their larger capacity filtering jugs are the best solution for households, where the needs are bigger and the water is needed, for example, for cooking or household appliances. Read more on Dafi.us
  20. High-quality reference and de novo genomes have been celebrated by geneticists, population biologists and conservationists alike, but it’s been a dream deferred for entomologists and others grappling with limited DNA samples, due to previous relatively high DNA input requirements (~5 μg for standard library protocol). A new low-input protocol now makes it possible to create high-quality de novo genome assemblies from just 100 ng of starting genomic DNA, without the need for time-consuming inbreeding or pooling strategies. The targeted release date for the protocol is February 2019. The protocol, developed as a collaboration by scientists at the Wellcome Sanger Institute and PacBio, was used to assemble the genome of an Anopheles coluzzii mosquito with unamplified DNA from a single individual female insect. As described in a bioXriv pre-print, Sarah B. Kingan, Haynes Heaton, et al. used a modified SMRTbell library construction protocol without DNA shearing and size selection to facilitate the use of lower input amounts, as shearing and clean up steps typically lead to loss of DNA material. “This new low-input approach puts PacBio-based assemblies in reach for small and highly heterozygous organisms that comprise much of the diversity of life,” said co-corresponding author Jonas Korlach, our chief scientific officer. The sample was run on the Sequel System with the latest v6.0 software, followed by de novo genome assembly with FALCON-Unzip, resulting in a highly continuous (contig N50 3.5 Mb) and complete (more than 98% of conserved genes were present and full-length) genome assembly. About a third of the new de novo genome is haplotype-resolved and represented as two separate sequences for the two alleles, providing additional information about the extent and structure of heterozygosity that was not available in previous assemblies, all of which were constructed from many pooled individuals. “The ability to generate high-quality genomes from single individuals greatly simplifies the assembly process and interpretation, and will allow far clearer lineage and evolutionary conclusions from the sequencing of members of different populations and species,” the authors state. The first Anopheles gambiae genome, published in 2002, was created using BACs and Sanger sequencing. Further work over the years to order and orient contigs improved this reference and to date, AgamP4 remains the highest quality Anopheles genome among the 21 that have now been sequenced. However, AgamP4 still has 6,302 gaps of Ns in the primary chromosome scaffolds and a large bin of unplaced contigs known as the “UNKN” (unknown) chromosome. The Sanger/PacBio single-insect assembly was able to place 667 (>90%) of the genes on the UNKN contigs into their appropriate chromosomal contexts. The assembly’s “gap-less mega-base scale contiguity” will also provide insights into promoters, enhancers, repeat elements, large-scale structural variation relative to other species, and many other aspects relative to functional and comparative genomics questions, the authors state. The protocol’s potential could also extend to other areas with typically low DNA input regimes, such as metagenomic community characterizations of small biofilms, DNA isolated from needle biopsy samples, and minimization of amplification cycles for targeted or single-cell sequencing applications, the authors add. Read more:https://www.cd-genomics.com/PacBio-SMRT-Sequencing.html
  21. Now that you have decided to build your dream home, you may have taken bold steps to locate the ideal lot or acreage site to build on. The impressive natural beauty of the land may have drawn you to it and instilled a sense of peace as you walked the lot. Understandably, you want to retain these same natural features that drew you to the land. Each lot and building project are unique, but these are some of the strategies that can be used to preserve the natural features that you have fallen in love with. Plan around Trees There are few natural features more beautiful than a centuries-old tree that has lush, healthy branches as well as deep roots and a huge trunk. These heritage trees tell a story of their own, so you understandably want to keep them intact on your land. If space permits, consider designing your home’s layout around the trees. These trees can be a true focal point to the landscape, and they add curb appeal as well as property value. Creating a floorplan and layout that takes the preservation of heritage trees into account is a smart idea. Build into Cliffs and Ledge If your lot has a dominant cliff or ledge, you may be able to build right up to the rock wall or even into it in some cases. Rock boring hire is available to help you anchor your property into the rock. By doing so, your home may become a part of the natural beauty. The rock may also act as a natural protective feature, and it could promote energy efficiency. It makes sense take full advantage of cliffs and ledges when they are present on your property. Hire an Experienced Architect These are only a few of the many features that you may want to preserve or take advantage of in your floorplan. In some cases, such as if you have a very large estate lot with ample space to lay the home on the land, special effort may not be required to preserve coveted natural features. However, other properties may require bold or monumental efforts in order to achieve your preservation goals. Hiring an experienced architect who has a proven track record of creativity and success in this area is advantageous. As you see, there are many creative ways to preserve your land’s treasured natural features. Identify the features that are most important to you, and speak with various contractors about the preservation options available.
  22. What does coating nanoparticles mean? As the name suggests, the process of applying a substance to the surface of a particle to modify it to form a composite particle is called coating of the nanoparticle. It has been found that this process can achieve uniform dispersion between different phase particles, and fully exert the excellent characteristics of different phase particles, and use it as a reinforcing particle. Because aluminum, iron-based, ceramic-based materials can have excellent properties such as high specific strength, wear resistance, small thermal expansion coefficient, and low cost, coating nanoparticles are widely used in aerospace, aviation, automotive, chemical, electronics, etc. A typical representative of coated nanoparticles Magnetic nanoparticles are nano-sized particles, generally composed of a magnetic core composed of a metal oxide such as iron, cobalt, or nickel, and a high molecular polymer/silicon/hydroxyapatite shell layer wrapped around a magnetic core. The most common core layer is made of Fe3O4 or γ-Fe2O3 with superparamagnetic or ferromagnetic properties. It has magnetic orientation (targeting). Under the action of external magnetic field, it can realize directional movement, convenient positioning and separation from medium. . Among them, the most common one is peg coated iron oxide nanoparticles. Several satisfactory research results Nanomaterials themselves exhibit different properties from conventional materials in terms of optical, thermal, electrical, magnetic, mechanical, physical, and chemical properties due to quantum size effects, surface effects, and macroscopic quantum tunneling effects that conventional materials do not possess. Moreover, the addition of nanoparticles can also improve the performance of traditional organic coatings. This topic has become a hot spot in the field of organic coatings research, and has obtained some satisfactory results: Using the electromagnetic and optical properties of the nanoparticles to produce a multifunctional coating with stealth function; The use of nanoparticles has a stronger flop effect, resulting in a more decorative coating for luxury car topcoats; The addition of nanoparticles can also improve the aging resistance and protective properties of the coating; Utilizing the absorption of ultraviolet rays by titania nanoparticles to form an ultraviolet shielding coating; In the field of anti-corrosion coatings, in addition to improving the performance of the coating by adding nanoparticles, the nanoparticles can be further modified by the nanoparticles to improve the protection of the material. The process of coating nanoparticles (taking SiC as an example) Non-uniform nucleation The principle is to use the coated particles as a nucleation matrix to control the concentration of the coating material between the critical value required for non-uniform nucleation and the critical value required for uniform nucleation, so that the nucleus of the coating material grows on the surface of the coated particles. The key to the process is to control the concentration of the appropriate coating material precipitation reaction and maintain the stable suspension characteristics of the coated particles. Problems to be solved At present, the research and application of SiC surface modification and coating at home and abroad are still in the initial stage. Due to various conditions, many related problems have not been solved, such as: Surface modification of SiC nanoparticles, optimum process conditions for coating process, and modification and coating mechanism; Development of low-phosphorus plating solution with low energy consumption, less pollution and renewable utilization; Uniform dispersion of SiC nanoparticles in the plating solution and in the coating layer; The behavior and mechanism of SiC nanoparticles in the coating layer; At present, the processes used at home and abroad are relatively high in cost, and require high-pressure, high-temperature conditions and high-purity SiC nanoparticles, or complicated operation, many process flows, resulting in low yield or uneven coating, and is not strong. The quality of the product is not good. In summary, reducing costs, simplifying steps, and ensuring quality will be an effort in the future. Latest results: to improve electrochemical performance of electrode materials At present, a Chinese R&D team has chosen to use nanoparticle surface coatings to improve the performance of lithium-rich electrodes. The team constructed a carbon shell of NiCo nanoparticles on the prepared nanoparticles, and the resulting LLO@C&NiCo cathode exhibited excellent cycle and rate performance. Analysis shows that LLO@C&NiCo, as the cathode material of lithium battery, exhibits excellent electrochemical performance due to its protective C&NiCo shell. This research has been supported by China's national key research and development plan and national key basic research development plan.
  23. Do you understand these important concepts? Nanoparticles refer to particles having a particle size between 1 and 100 nm, also known as ultrafine particles. They are in an over-extension between macroscopic objects and between micro-systems and macro-systems. They are a group of a small number of atoms or molecules, so they are neither a typical microscopic system nor an atypical macroscopic system. The basic meaning of nanoscience and technology is to manipulate and arrange atomic and molecular innovations directly or indirectly in the nanometer category for making new substances. Nanomedicine is an emerging edge interdisciplinary subject based on nanoscience or medicine. Its basic meaning refers to the use of nanotechnology theories and methods to carry out a new discipline of medical research and clinical treatment, which is mainly related to medicine in two aspects. Combination: First, the detection technology of disease; second, the treatment technology of disease. As an important tool in the field of nanomedicine, gold nanoparticles have become a research hotspot in the field of nanomedicine at home and abroad with its unique properties. The emergence of nanoparticles has injected new impetus into the scientific development of the new century! Preparation of nanoparticles Ø Vapor Deposition: Synthesis of nanomaterials by chemical reaction of metal compound vapors. It is characterized by high purity and narrow particle size distribution. Ø Precipitation method: After the precipitant is added to the salt solution for reaction, the precipitate is heat-treated to obtain a nano material. Its characteristics are simple and easy, but the purity is low and the particle radius is large. Ø Hydrothermal synthesis method: synthesis in a fluid such as an aqueous solution or a vapor under high temperature and high pressure, and then separating and heat-treating to obtain nanoparticles. It is characterized by high purity, good dispersibility and easy control of particle size. Ø Sol-gel method: a metal compound is solidified by a solution, a sol or a gel, and then subjected to a low heat treatment to form a nanoparticle. It is characterized by a large number of reaction species, uniform product particles, and easy process control. Ø Microemulsion method: two mutually incompatible solvents form an emulsion under the action of a surfactant, and nucleation, coalescence, agglomeration and heat treatment in the microbubbles to obtain nanoparticles. It is characterized by monodisperse and good interface of particles. Nanoparticles are widely used in biology, chemistry, medicine, etc. Nanoparticle probe Bioanalysis is mainly used in the frontier fields of biological macromolecule analysis, biopharmaceutical analysis, bioactive substance analysis and microbial analysis such as proteins and nucleic acids. Because the sensitivity of the existing methods to the specificity of the target molecule is not high, scientists have sought to find a more sensitive and specific method for bioanalysis. The emergence of nanoparticle probes provides a new way to solve this problem. It can accurately combine the target and has strong specificity. At the same time, the nanometer size of the probe can significantly improve the sensitivity, and the in-depth study of biomolecules. Convenience is provided. Classification of nanoparticle probes: ² semiconductor nanoparticle probes ² metal nanoparticle probes ² composite nanoparticle probes ² magnetic nanoparticle probes Applications of nanoparticle probes: ² DNA detection ² protein detection ² drug targeted delivery u Diagnosis of malignant tumors As a new type of nanocarrier, mesoporous silica nanoparticles have a good application prospect in the field of biomedicine. It is different from the physicochemical properties of traditional inorganic materials and plays a key role in the diagnosis and treatment of malignant tumors. In particular, MSNs, as a drug-loading platform with high loading capacity, good biocompatibility, targeting and controllability for drug release, can be used to solve the problems encountered in the current clinical diagnosis and treatment of malignant tumors. Nano pesticide Nanotechnology is a highly attractive research area that is emerging to achieve these goals. It is a new approach to designing novel nano-sized active ingredients and their formulations and delivery systems, collectively referred to as "nano-pesticides." Nanopesticide is an emerging field called crop nanotechnology for crop protection. This field covers a wide range, including a basic understanding of the interaction between nanomaterials and insects, making existing pesticide active ingredients into nanoemulsions and dispersants, using nanomaterials as active pesticide ingredients or developing new nanopesticide formulations using nanomaterials as carriers. It is expected that research on nanopesticide will address the major limitations of existing pest management strategies, providing new and advanced nano-formulations that are stable and active in the target environment (less affected by sunlight, heat and rain) and can penetrate Target organisms (insects), which are resistant to pest defense mechanisms, are harmless to plants and mammals, are cost-effective to formulate and produce, and have new mechanisms of action. Common carriers for pesticide delivery systems: ² Nano polymer particles ² inorganic nanoparticles ² silver nanoparticles ² other mineral nanoparticles Nanotechnology can reduce the use of crop protection chemicals and will make agriculture environmentally friendly and cost effective. Fertilizers, pesticides, and growth regulators are well-delivered, including nanosensors that detect soil conditions, crop growth, and pest and disease infestation in time, through the development of nanodevices and products. The application of nanotechnology in agriculture will have good prospects as well as in other fields, but progress will be relatively slow.
  24. In cities across the country and beyond, there is an increasing push to protect the planet from additional harm and hope that some damage can be reversed. Many cities are taking bold steps to be more environmentally-friendly in various ways, and these efforts may inspire you to make positive changes as well. These are some of the more common ways cities are becoming greener. Water Conservation There have been several recent news stories about smaller and larger cities around the world running low on fresh water, so water conservation is understandably a hot topic. Many cities are taking bold steps to conserve water, such as by turning water in a sewage system into sanitized, healthy drinking water. Some cities are collecting rainwater to use for landscape watering, enforcing water conservation laws and more. Recycling Programs Government recycling programs have been around for decades, but some cities are much more proactive in this area than they have been in the past. For example, many cities now offer curbside recycling service as well as special collection days for appliances and large items. There are recycling programs for Christmas trees and even special warehouses where old building supplies may be donated for use by others. In government buildings, employees may be required to participate in recycling programs. Energy Reduction Another common step that cities are taking to go green involves energy consumption. Increasingly, you will find solar panels on government buildings so that the buildings can generate at least some of the electric power that they need. City buses may likewise run on natural gas or even have electric engines. In addition to these steps, there may be a push to reduce energy consumption, such as through an investment in equipment that uses less energy. These are only some of the many ways that cities today are going green or becoming greener than they already were. As you explore ways to go green at home, you may look at your city’s efforts for inspiration. For example, you can trade in your car for an electric or hybrid vehicle, and you could install solar panels on your home. You may increase your participation in a recycling program and decrease personal use of electricity and water. While you could make drastic changes right away, even making small improvements regularly can have a huge impact on the planet’s health over time. Look for ways that you can decrease your impact on the environment today, and take action soon.
  25. Whole exome sequencing (WES), also named as exome sequencing, is a genomic technique to sequence all of the protein-coding genes in exome. It contains two parts: the first is to select only the subset of DNA that encodes proteins, known as exons. The second is to sequence the exonic DNA by any high-throughput DNA sequencing technology. By using this technique, fixed-cost studies can sequence samples to much higher depth than that of whole genome sequencing. This additional depth makes exome sequencing well suited to several applications that need reliable variant calls, for example, rare variant mapping in complex disorders, discovery of Mendelian disorders, case studies, clinical diagnostics and direct-to-consumer exome sequencing. In this article, we listed 20 frequently asked questions about whole exome sequencing. 1. Does the whole exome sequencing have a reference genome? Yes, there should be a sequence of related species if there is no reference group, but the reliability of the capture results is not guaranteed. Since the capture probe is designed based on the provided reference sequence, it is not recommended if the target region is known to have a large discrepancy with the reference genome, such as a large fragment insertion. 2. Why does exon sequencing need to be compared with the whole genome in the analysis, rather than directly compared to the target area? First, the target area is generally shorter than the whole genome, and may be discontinuous. If the target area is extracted separately, it will affect the sequence alignment effect at the edge of the area; Second, the quality of the capture cannot be assessed, such as off-target rate, on target ratio, and the like. 3. How many coverage times are preferable for whole exome sequencing? Generally, 100 × or 150 × is recommended. With a higher coverage, some rare mutations is easier to be found for the genetic deterioration of heterogeneity. In addition, the coverage of exon sequencing is random, so a higher average coverage is beneficial to ensure that most areas have sufficient coverage. 4. What is the significance of whole exome sequencing depth? How is the sequencing depth converted? The sequencing depth represents the number of times of the sequence is covered by the probe set. The higher the number, the more accurate the sequencing result is, and the more accurate the subsequent statistical analysis. If you run tumor, low frequency mutation studies, it is recommended that the sequencing depth should be at least 150 ×. If you only look for classic SNPs, non-low frequency mutations, the sequencing depth should be at least 30×. Sequencing depth conversion method: The capture efficiency of the general target area is 60-70%, and the target area of the exon capture kits such as Agilent and Roche are about 60 Mb, that is, the sequencing depth = 10G*60%/60Mb=100×. 5. How many missing fragments can whole exome sequencing detect? A 50 bp fragment deletion can be roughly measured. Because the coverage of exon sequencing is very uneven, if there is a large segment of deletion, it cannot be judged whether the hybridization is not captured or it is missing. What is currently detectable is a missing found in a read. The length of a read is about 150 bp, so fragment deletions below 50 bp can be detected from exon sequencing. 6. Can whole exome sequencing be used in CNV analysis? What are the methods for detecting CNV? Whole exon sequencing has a hybrid capture efficiency problem because of a hybrid capture process. The hybridization efficiency of each exon is different, and the homologous competition is different, so the coverage difference of different exons is very large. Therefore, in general, exon sequencing cannot be used for CNV detection. However, in cancer research, CNV can be detected using cancer tissue and paracancerous tissue controls. There are two conventional methods for detecting CNV, one is whole genome resequencing and the other is Affymetrix SNP 6.0. Among them, Affymetrix SNP6.0 has a relatively low detection cost and is a relatively economical means. 7. Can whole exome sequencing be performed in the target region for methylation detection? Can RNA capture be performed? Target region methylation assays can be directly used with methylation capture sequencing, such as SureSelect's MethylSeq or NimbleGen's SeqCap Epi. The principle is similar to capture sequencing, which is using specialized probes to capture and enrich target area. The target region is subjected to capture enrichment, sequencing and methylation detection after bisulfite treatment. RNA capture can be performed using SureSelect's RNA Capture or NimbleGen's SeqCapRNA System. 8. Can capture samples be mixed with multiple species? Yes, as long as the captured target fragment has a corresponding reference genome, it can be captured by the probe. For example, viral sequences integrated in the host genome, mixed parasite sequences in blood, low abundance microbial sequences in environmental samples, and the like. Since the proportion of these sequences in the mixed sample is often very low, the efficiency of using the traditional resequencing method will be very low, and a very high amount of sequencing is required to obtain sufficient coverage depth, and a large amount of redundant data is generated. In such cases, capture sequencing has an absolute advantage over them. 9. Is there any indicator for the effect of the capture? What factors will reduce the capture effect? Due to the limitation of sequencing technology, there are some repeating sequences, undetermined “N bases”, and the quality of the sample itself, which may cause the sequence to be uncovered. This is a problem that all sequencing technologies will encounter. Deep genome sequencing does not guarantee 100% complete coverage of the genome. Since each sample varies widely, it is not reliable to make a direct commitment to the capture effect. The best way is: client provides target area, and company designs and evaluates them to get detailed reports. In addition, there are ubiquitous repeat sequences in the species. Before designing, it is generally necessary to shield some difficult-to-cover repetitive masks and then design them, which will improve the quality of the capture, but at the same time reduce the overall coverage. 10. Can high GC content fragments be captured? Yes, but there will be some impact on the capture efficiency of these fragments. Similarly, low complexity segments and containing fuzzy base segments have some difficulty in capturing. When designing the probe, company technicians appropriately increase the number of coverages and probe density according to the coverage, and send the expected capture effect to the customer for confirmation. In addition, if you are capturing full exons, UTR, etc., you can use preset probe sets. These probe designs are optimized, which improves the corresponding capture efficiency. 20 Frequently asked Questions (FAQs) about Whole Exome Sequencing (Part Two) 11. What is the efficiency of exon capture (whole exome sequencing)? The hybridization process is used during exon sequencing. There are many parts of the human chromosome that are homologous to exons, and these homologous parts are likely to be captured during the hybridization process. Therefore, some of the sequences detected are not exon sequences. We refer to the ratio of the sequence of exons to the entire sequencing sequence as the capture efficiency. The efficiency of capture does not affect the quality of the data, but the effective proportion of the data. 12. When do you choose whole exon capture? When do you choose custom probe capture? All exon captures of humans and some common species have pre-set probe products, of which human all-exon probes have been optimized for many times. If there are many target regions and all are exons (several tens of Mb or more), it is recommended to select whole exon capture directly, because the whole exon has a more mature probe set and lower cost, and the capture effect is better. Custom probes are recommended for smaller segments of interest, or for segments other than exons. 13. What information are needed for custom design exon capture probe? Species, reference genomic information, target area information (coordinates, gene names or gene numbers, etc.) are needed. If only coordinate information is provided, it should be provided in the form of a table. If the gene name is provided, it is best to have the official name of NCBI and the corresponding gene number. Multiple target areas may overlap without affecting subsequent design and analysis. After the design is completed, the final regional information will be returned to the customer for confirmation. The target area supports a maximum of 200Mb. After providing the gene name, company will help customers find specific regions of the gene, such as exons, UTR, and so on. If you want to capture several or all of them, you need to indicate. 14. Can the conventional species or new species genomes, imperfect or much-error genome be used for exon capture sequencing? Yes, the capture sequencing platform has no species restriction, as long as genomic sequence information and target region information are available. Capture sequencing design can be performed even if the reference genome of the species is new, inaccurate, or even very different. Correspondingly, the captured probes are designed according to the sequence provided by the customer, so the accuracy of the results cannot be fully guaranteed. 15. Can the degradation sample be used for post-library construction? What is the impact on subsequent data and analysis? If the degradation of the sample is very serious, it is not recommended to build the database. The success rate of the database is low. If the sample is seriously degraded, the impact of constructing the exon library will be low, the effective data volume is low, and the properly mapped will be low. 16. What is the impact of viscosity sample, pore impurity contamination and RNA contamination on library construction? What is the success rate? Slight protein contamination has little effect on the library construction, but if the protein or other impurities are seriously polluted, it will affect the quantification, and the enzyme efficiency in the database, which will reduce the success rate of the database; RNA pollution mainly affects the sample quantification and the sorting of DNA in the database. Therefore, if there is RNA contamination, RNA digestion is recommended when the total amount and quality of the sample are appropriate. 17. Can a tissue be co-extracted with DNA and RNA? What is the extraction method? Co-extraction can be performed; there are generally two methods for co-extraction, one is to divide the tissue into two parts, each for DNA and RNA extraction; the other is to use the DNA/RNA co-extraction kit for extraction, but generally degradation of DNA and RNA is likely to happy after extraction. 18. What is the reason for the low DNA extraction yield of FFPE samples? FFPE samples are generally stored for many years, and they have been seriously degraded by formaldehyde; and FFPE samples are generally precious, and the number of samples sent is less. 19. What is the difference between cfDNA and ctDNA? cfDNA (cell free DNA) is a free DNA in the blood. It is released from the normal cells of the body or the rupture of white blood cells. It is harmless to the body and will soon be cleaned by itself. ctDNA (circulating tumor DNA) is the rupture of tumor cells. Free DNA released into the blood can be used as a highly specific tumor marker. 20. How to separate plasma samples when extracting cf/ctDNA? Collect 5 mL of peripheral blood (usually in the morning), quickly transfer to the EDTA anticoagulant tube, carefully invert and mix (prevent hemolysis); run internal plasma separation within 1 hour (at room temperature) or 2 hours (at 4 °C): Centrifuge at 1000 rpm for 10 minutes at 4 °C, carefully pipette the plasma to a clean 1.5 mL EP tube (not to suck white blood cells during pipetting), then centrifuge at 12,000 rpm for 10 minutes at 4 °C to remove residual cells or debris, and carefully absorb the required supernatant volume into a new dispensing EP tube, mark with an oily marker, and store in an ultra-low refrigerator at -80 °C to avoid repeated freezing and thawing, and transport samples with dry ice.
  26. About Plasmid What is a plasmid? A plasmid is a genetic unit capable of autonomous replication outside the chromosome, including organelles of eukaryotes and deoxyribonucleic acid (DNA) molecules except for chromosomes in bacterial cells. It is now customary to refer to DNA molecules in bacteria, yeasts, and actinomycetes other than chromosomes. In genetic engineering, plasmids are often used as vectors for genes. Many bacteria have a large number of small circular DNA molecules in addition to chromosomes. This is a plasmid (some plasmids are RNA). The plasmid often has antibiotic resistance genes, such as tetracycline resistance gene or kanamycin resistance gene. Some plasmids are called episomes, and these plasmids can be integrated into the chromosome of the bacteria. From the integration position, it becomes a DNA molecule that is free from extrachromosomes. Plasmid categories At present, hundreds of bacteria have been found to have plasmids. Most of the known bacterial plasmids are closed circular DNA molecules (cccDNA). The relative molecular mass of bacterial plasmids is generally small, about 0.5%~3% of bacterial chromosomes. According to the relative molecular mass, plasmid can be roughly divided into two types: the relative larger one is 40×106 or more, and the relative smaller one is 10×106 or less. (A few molecular masses of plasmids are somewhere in between.) The number of plasmids in each cell is mainly determined by the replication characteristics of the plasmid itself. According to the nature of replication, the plasmids can be divided into two categories: one is a tight-type plasmid. When the cell chromosome is replicated once, the plasmid is also replicated once, and there are only 1 or 2 plasmids in each cell; the other is a relaxed plasmid, which can continue to replicate when the chromosome replication is stopped. There are generally about 20 plasmids in each cell. The replication of these plasmids is under the control of the relaxation of the host cells, containing 10–200 copies per cell. If a certain drug treatment inhibits the synthesis of the host protein, the plasmid copy number is increased to several thousand. The earlier plasmid pBR322 belongs to the relaxation plasmid, which can be treated with chloramphenicol to reach higher copy number. Generally, the plasmid with larger molecular weight is tight-type. The plasmid with smaller molecular weight is relaxed-type. The replication of plasmids sometimes is related to their Host cell. Plasmid application In genetic engineering, artificially constructed plasmids are commonly used as vectors. Artificially constructed plasmids can integrate a variety of useful features, such as a variety of single enzyme cleavage sites, antibiotic resistance, etc. Commonly used artificial plasmid carriers have pBR322 and pSC101. pBR322 contains anti-tetracycline gene (Tcr) and ampicillin resistance gene (Apr), and contains a single point of 5 endonucleases. If the DNA fragment is inserted into the EcoRI cut point, it will not affect the two antibiotic genes expression. However, if the DNA fragment is inserted into the Hind III, Bam HI or Sal I point, the anti-tetracycline gene will be inactivated. At this time, the pBR322 containing the DNA insert will make the host bacterium resistant to ampicillin but sensitive to tetracycline. pBR322 without a DNA insert would make the host bacterium resistant to both ampicillin and tetracycline, whereas bacteria without the pBR322 plasmid would be sensitive to ampicillin and tetracycline. pSC101 is similar to pBR322 except that it has no anti-ampicillin gene and PstI nicking. The largest insert of the plasmid vector is approximately 10 kb (kb is expressed as kilobase pairs). Plasmid sequencing Before starting a plasmid sequencing project, one should consider: The DNA quality and purity Is there host DNA contamination? Sole plasmid or genome and plasmid? Plasmid sequencing Applications Ÿ Mutation studies Ÿ Vector verifications Ÿ Characterization of production strains CD Genomics provides complete plasmid DNA sequencing service, and our improved bioinformatics pipelines are available to perform de novo assembly with no reference required.
  27. CD Genomics, the USA-based genetic sequencing company, recently announces the launch of its proprietary GenSeqTM Technology to provide comprehensive Single-Cell Sequencing services. We already knew that a typical human cell consists of around 2 x 3.3 billion base pairs of DNA and 600 million bases of mRNA. In normal condition, a mix of millions of cells are used during the course of sequencing the DNA or RNA with traditional methods (such as Sanger sequencing/ Illumina sequencing). With such deep sequencing of DNA and RNA from a single cell, the cellular functions can be investigated through-fully. Under the setting of rapid progress in the development of next-generation sequencing (NGS) technologies, this field has provided many valuable insights into complex biological systems and dramatically advanced cell biology, for example, from cancer genomics to diverse microbial identification. It has been treated as a promising tool for approaching a set of seemingly inaccessible problems. For example, heterogeneous samples, rare cell types, cell lineage relationships, mosaicism of somatic tissues, analyses of microbes that cannot be cultured, and disease evolution can all be elucidated through single cell sequencing. With such progress, researchers are now increasingly moving their focus from whole to the characterization of individual cells-based technologies for genomics, transcriptomics, and epigenomics. “These single-cell analyses will allow researchers to uncover new and potentially unexpected biological discoveries relative to traditional profiling methods that assess bulk populations. Single-cell RNA sequencing (scRNA-seq), for example, can reveal complex and rare cell populations, uncover regulatory relationships between genes, and track the trajectories of distinct cell lineages in development”, as said by Byungjin Hwang. Single-cell sequencing could be applied in: Profiling scarce clinical samples Measuring intra-tumor heterogeneity and guiding chemotherapy Cancer cells evolution analysis during tumor progression Pre-implantation genetic diagnosis (PGD) Similar to NGS experiments, single cell sequencing protocols generally cover the following four steps: isolation of a single cell, nucleic acid extraction and amplification, sequencing library preparation, sequencing and bioinformatic data analysis. About CD Genomics Single-Cell Sequencing services CD Genomics’ Single-Cell kit produces amplified DNA fragments suitable for Copy Number Variation (CNV) analysis using oligonucleotide aCGH or qPCR; SNP genotyping, mutation detection and sequencing. Advantages of CD Genomics Single-Cell Sequencing Complete: End-to-end workflow for whole transcriptome analysis of individual cells. Highest throughput: Unprecedented parallel processing of up to 96 single cells per run. Easiest to use: Less than three hours total hands-on time, working directly from single cells, with no RNA fragmentation and purification step. Affordable: One-eighth the cost of other library preparation system.
  28. The signal pathway means that when a certain reaction occurs in a cell, the signal transmits a message from outside the cell to the cell, and the cell responds according to the information. The signal pathway was first traced back to 1972, but it was called signal transmission at that time. In 1980, M. Rodbell referred to signal transduction in a review, and the concept has since been widely used. A signaling pathway is a series of enzymatic reaction pathways that can exert extracellular molecular signals through the cell membrane and into cells. These extracellular molecular signals (called ligands) include hormones, growth factors, cytokines, neurotransmitters, and other small molecule compounds. Definition How does the signal in the cell pass when the ligand specifically binds to the receptor in the cell membrane or cell? The various biochemical pathways in the cell are composed of a series of different proteins that perform different physiological and biochemical functions. The regulation of downstream protein activity (including activation or inhibition) by upstream proteins in each signaling pathway is primarily accomplished by the addition or removal of phosphate groups, thereby altering the stereoconfiguration of downstream proteins. Therefore, the major members of the signaling pathway are protein kinases and phosphatases, which are capable of rapidly altering and restoring the conformation of downstream proteins. Receiving an external signal from a cell receptor to finally making a comprehensive response is not only a signal transduction process, but more importantly, a process of gradually amplifying the external signal. Receptor proteins convert extracellular signals into intracellular signals, which are amplified, dispersed, and regulated by signal cascades, resulting in a comprehensive array of cellular responses, including regulation of downstream gene expression, changes in intracellular enzyme activity, and cellular bone architecture. Type and changes in DNA synthesis, etc. These changes are not all caused by one type of signal, but different reactions can be produced by different combinations of several signals. Classification First, when the signal molecules are lipids such as cholesterol, they can easily cross the cell membrane and bind to the target receptor in the cytoplasm; Second, when the signal molecules are polypeptides, they can only bind to receptors such as proteins on the cell membrane. These receptors are mostly transmembrane proteins. Through conformational changes, signals are transmitted from the extracellular domain to the domain within the membrane, and then Acting with the next level of receptors, the next level of pathway is activated by modification such as phosphorylation. Common signal path NF-κB signal NF-kB (nuclear factor-kappa is a transcription factor found in the nuclear extract of B lymphocytes in 1986. It binds specifically to the enhancer B sequence GGGACTTTCC of the immunoglobulin kappa light chain gene and promotes κ. Light chain gene expression, hence the name. It is a member of the Rel family of eukaryotic transcription factors and is widely found in various mammalian cells [1]. To date, five NF-kB/Rel family members have been found in mammalian cells, which are RelA (ie, p65), RelB, C-Rel, p50/NF-kB1 (ie, p50/RelA) and p52/NF. -kB2. These members all have a Rel homology domain (RHD) of approximately 300 amino acids. This highly conserved domain mediates the formation of a homologous or heterodimer of the Rel protein, which is also a region of specific binding of NF-kB to the DNA sequence of the target gene. The activation process of NF-kB in cells is finely regulated. Normally, NF-kB in the cytoplasm is inactivated and binds to the inhibitory protein of NF-kB to form a trimer complex. When TNF-a signaling, inflammatory factors, and external stimuli such as LPS and ultraviolet rays are present, cytokines bind to TNF receptors on the surface of cell membranes, and TNF receptors multimerize and interact with TRADD molecules in the cytoplasm. TRADD recruits TRAF (TNFR-associated factor) and kinase RIP (receptor interacting protein), and signals are transmitted by RIP to IKK (IkB kinase). IKK plays a very important role in the NF-kB signaling pathway, and although it is different in the upstream signal path, it eventually aggregates into IKK. IKK consists of three subunits a, b and g. IKK as a kinase phosphorylates the Ser32 and Ser36 residues of the a subunit of IkB and the Ser19 and Ser23 residues of the b subunit. IkB is then dissociated from the p50/p65/IkB heterotrimer and degraded by proteasome after ubiquitination. Thus, NF-kB, which is inhibited by IkB, is exposed to its nuclear localization signals (NLS), rapidly entering the nucleus from the cytoplasm, and binding to specific sequences on the DNA in the nucleus to initiate or enhance transcription of related genes. 2. JAK-STAT signaling pathway 1) JAK and STAT proteins The JAK-STAT signaling pathway is a signal transduction pathway stimulated by cytokines discovered in recent years, and is involved in many important biological processes such as cell proliferation, differentiation, apoptosis and immune regulation. Compared with other signaling pathways, this signaling pathway is relatively simple to transfer. It consists of three components, namely tyrosine kinase-related receptor, tyrosine kinase JAK and transcription factor STAT. 2) JAK-STAT signaling pathway The transfer of the JAK-STAT signaling pathway is relatively simple compared to other signaling pathways. The signaling process is as follows: Binding of cytokines to the corresponding receptor causes dimerization of the receptor molecule, which allows the JAK kinases coupled to the receptor to be close to each other and activated by interactive tyrosine phosphorylation. After JAK activation, the tyrosine residues on the catalytic receptor are phosphorylated, and then these phosphorylated tyrosine sites form a "docking site" with the surrounding amino acid sequence, and contain the SH2 domain. STAT protein was recruited to this "parking site." Finally, the kinase JAK catalyzes the phosphorylation of the STAT protein bound to the receptor, and the activated STAT protein enters the nucleus in the form of a dimer to bind to the target gene and regulate gene transcription. It is worth mentioning that a JAK kinase can be involved in the signal transduction of many cytokines. A cytokine signaling pathway can also activate multiple JAK kinases, but cytokines have certain choices for activated STAT molecules. Sex. For example, IL-4 activates STAT6, while IL-12 specifically activates STAT4. 3.Ras, PI (3)K and mTOR signals With the completion of the sequencing of the human genome, hundreds of protein kinases have been discovered. Based on their structural similarities, these kinases can be divided into multiple protein families that play important biological functions in cell proliferation, growth, differentiation, and apoptosis. Ras, PI(3)K and mTOR are a class of protein kinases closely related to cell proliferation. The normal growth of eukaryotic cells is limited by the nutrients provided by the surrounding environment. Ras and PI (3)K signaling play a key role in regulating cell growth by regulating the downstream molecule mTOR. In most human tumor cells, key mutations in the Ras and PI (3)K signaling pathways have undergone significant mutations. The reason is that if this signal pathway is mutated, it will lead to cell survival and growth no longer limited by environmental conditions such as nutrients, and then induce cell cancer. It is worth noting that some tumor suppressors, such as TSC1, TSC2 and LKB1, attenuate the intensity of the mTOR signaling pathway under nutrient-deficient conditions. Correspondingly, inactivation mutations in TSC1, TSC2 or LKB1 result in similar cancer symptoms with a common clinical manifestation. Therefore, this signal pathway that ensures that cells proliferate under environmentally appropriate conditions can be used by cancer cells to survive and grow under conditions of lack of nutrients. In the process of screening for kinase inhibitors, a series of drugs targeting kinases such as mTOR, PI (3)K, RTKs and Raf have been designed. In the molecular mechanism of cancer research, although this signaling pathway is the most thoroughly studied, the physiological functions of these kinases in cells and organisms are far more complicated than we think. 4.Wnt signal The Wnt signaling pathway is widely distributed in invertebrates and vertebrates and is a highly conserved signaling pathway during species evolution. Wnt signaling plays a crucial role in the early development of animal embryos, organ formation, tissue regeneration and other physiological processes. If a key protein in this signaling pathway is mutated, resulting in abnormal activation of the signal, it may induce cancer. In 1982, R. Nusse and H.E. Varmus cloned the first Wnt gene in mouse breast cancer cells, initially named Int1 (integration 1). Later studies found that the mouse Int gene and the wingless gene wg (wingless) of Drosophila are homologous genes, and thus the two are collectively referred to as Wnt. H.E. Varmus I also won the 1989 Nobel Prize in Physiology and Medicine for his outstanding contribution to cancer research. Wnt is a type of secreted glycoprotein that acts through autocrine or paracrine. The main components of the Wnt signaling pathway include: the secreted protein Wnt family, the transmembrane receptor Frizzled family, CK1, Deskerled, GSK3, APC, Axin, β-Catenin, and the transcription factor TCF/LEF family. The Wnt signaling pathway is a complex regulatory network that is currently thought to include three branches: the canonical Wnt signaling pathway, which activates gene transcription via β-Catenin; the Wnt/PCP pathway (planner cell polarity pathway), which activates JNK via small G proteins (c -Jun N-terminal kinase) regulates cytoskeletal rearrangement; the Wnt/Ca2+ pathway affects cell adhesion and related gene expression by releasing intracellular Ca2+. It is generally mentioned that the Wnt signaling pathway mainly refers to the classical Wnt signaling pathway mediated by β-Catenin. 5. BMP signaling pathway BMP (bone morphogenetic protein) is an important member of the TGF-β (transforming growth factor-β) superfamily [6]. By regulating the activity of a range of downstream genes, many important biological processes such as mesoderm formation, nervous system differentiation, tooth and bone development, and cancer development are controlled. The BMP signal is specifically transmitted by the ligand BMP to specifically bind to the serine/threonine kinase receptor (BMPR) on the cell membrane to form a ligand receptor binary complex. At the same time, the type II receptor (BMPR2) is able to activate the type I receptor (BMPR1) and further transmit signals to the Smad molecules in the cell. The Smad protein plays a key role in the transmission of BMP and TGF-β signals from the cell membrane to the nucleus. Activated type I receptor (BMPR1) further phosphorylates Smad proteins (Smad1, Smad5, and Smad8), causing Smad molecules to detach from cell membrane receptors and bind to Smad4 molecules (common Smad, Co-Smad) in the cytoplasm Enter the nucleus. In the nucleus, the Smad multi-component complex acts on a specific target gene with the participation of other DNA-binding proteins to regulate the transcription of the target gene. 6. Ras2MAPK signal transduction pathway 1) Ras upstream channel Ras can be activated by complex networks. First, phosphorylation-activated receptors, such as PDGFR, EGFR directly bind to growth factor receptor binding protein (Grb2), which also bind indirectly and phosphorylate proteins containing the src homology region 2 (SH2) domain (For example, Shc, Syp), then activate Grb2. In addition, the src homology 3 (SH3) domain of Grb2 binds to target proteins such as mSos1, mSos2, C3G and dynamin. The SH3 structure of C3G and connexin Crk Domain binding after coupling tyrosine phosphorylation to activate Ras. Crk can also bind mSos1 to activate Ras.Grb2 to bind to activated receptors to promote the localization of guanylate exchange factor (Sos) protein on the cell membrane adjacent to Ras. Sos forms a complex with Ras. After GTP replaces GDP and Ras, Ras is activated. When GTP is hydrolyzed into GDP, Ras is inactivated. Ras has intrinsic GTPase activity, and its activity can be regulated by RasGAPs, thus RasGAPs act as Ras activity regulation. The role of the agent. In addition, Ras inactivation is also highly regulated. Currently, there are three proteins that can hydrolyze GTP to inactivate Ras, which are P120GAP, neurofibromin and GAP1m, collectively referred to as RasGAPs. 2) Ras downstream path At present, the Ras/Raf pathway is the clearest signal transduction pathway. When GTP replaces GDP and Ras binds, Ras is activated, then activates the silrethine kinase cascade amplification effect, recruiting intracellular Raf1 serine kinase to the cell membrane, Raf kinase phosphorylates MAPK kinase (MAPKK), and MAPKK activates MAPK. MAPK is activated and then transferred to the nucleus to directly activate transcription factors. In addition, MAPK stimulates Fos, Jun transcription factor forms transcription factor AP1, which is related to the specific DNA sequence next to the myc gene binds to initiate transcription. The myc gene product is also a transcription factor that activates other genes. Ultimately, these signals are concentrated to induce the expression and activity of D-type Cyclin. D-type Cyclin and Cyclin-dependent kinases (such as CDK4 and CDK6) form a complex that forms the cell from the G1 phase to the S phase. Therefore, the Ras/Raf pathway plays a key role between the receptor signal and G1 phase progression, however, Ras/Raf Pathway is not the only pathway regulating G1 progression. Ras and Raf alone do not promote Raf kinase activity, while Raf can be activated by Ras-independent mechanisms (eg, activated by Src tyrosine kinase and PKC), MAPK Can be independent of Ra the s mechanism (eg, by modulating the activity of integrins) is activated. This indicates that each signaling protein in the cascade reaction can be activated by multiple upstream proteins, and they may have additional target proteins. About us We leverage our wide spectrum of business in the fields of development, manufacturing, marketing, and distribution to help you make best-informed decisions tailored to your evolving needs for premium chemicals. Our complete suite of CRO services spans the entire molecule development pipeline including Apoptosis Signaling Pathway, B Cell Receptor Signaling Pathway, CTLA4 Signaling Pathway, and EGFR Signaling Pathway.
  29. Although the problem of protecting the environment may seem like an insurmountable one, there are many simple steps that every person can take to make a small positive impact. If everyone begins to take measures to reduce carbon use and waste, saving the planet will become a much easier proposition. Here are four simple things you can do to help save the environment in your own small way. Walk and Bike More Driving is one of the most common ways in which practically everyone releases greenhouse gases into the atmosphere. By walking or riding your bike for short trips, you can considerably reduce the amount of carbon dioxide produced as a result of your transportation needs. As an added benefit, walking and biking are excellent forms of exercise, meaning you’ll get some great physical activity as well. Plant Trees on Your Property If you have a large enough piece of property, consider planting a few trees to help absorb carbon dioxide. A single tree can sequester a full ton of carbon over the first 40 years of its life. By planting several trees on your property, you can create a passive form of carbon sequestration that will continue for decades to come. Get into the Habit of Recycling One of the best ways to prevent unneeded waste from polluting land and water is to recycle as much of your garbage as you can. Plastics, glass and paper can easily be recycled by most municipal governments, meaning that you just have to sort them appropriately. Iron, steel, copper, and aluminum scrap metal recycling are usually handled by specialized processing centers, but it isn’t too difficult to find these facilities in most cities. By recycling more of what you use, you can prevent waste from ending up in landfills and keep extra energy from being expended in extracting new natural resources. Grow Some of Your Own Food One of the more difficult environmental problems to address is the fact that the agricultural techniques needed to supply food for more than 7 billion humans are extremely harmful to the planet. Forests are often destroyed to clear land, while herbicides, pesticides and fertilizers pollute groundwater reserves on massive farms. One thing you can do to partially reduce your impact in this area is to start an organic garden at home. Growing your own food with environmentally friendly methods will lower your carbon footprint and give you access to plenty of fresh, healthy produce. These are just a few of the many small actions you can take to play a role in saving the planet. You should also try to encourage friends and family members to do the same, as the effect of these small steps will grow as more and more people begin to take them.
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