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    Scientists at Indiana University and international collaborators have found a way to link two hormones into a single molecule, producing a more effective therapy with fewer side effects for potential use as treatment for obesity and related medical conditions.

    The studies were carried out in the laboratories of Richard DiMarchi, the Standiford H. Cox Distinguished Professor of Chemistry and the Linda & Jack Gill Chair in Biomolecular Sciences in the IU Bloomington College of Arts and Sciences, and of Matthias Tschöp, professor of medicine and director of the Institute of Diabetes and Obesity, Helmholtz Center Munich, Germany.

    Results were published online this week by the journal Nature Medicine.

    Researchers combined a peptide hormone from the digestive system, GLP-1, with the hormone estrogen and administered it to obese laboratory mice. While both GLP-1 and estrogen have demonstrated efficacy as therapy for obesity and adult-onset diabetes, the combination was more effective in producing weight loss and other beneficial results than using either compound on its own. And it produced fewer adverse effects, such as excessive tissue growth linked to tumor formation.

    "We find that combining the hormones as a single molecule dramatically enhanced their efficacy and their safety," DiMarchi said. "The combination improves the ability to lower body weight and the ability to manage glucose, and it does so without showing the hallmark toxicities associated with estrogen."

    The researchers believe GLP-1 acts as a "medicinal chaperone," targeting estrogen to the hypothalamus and pancreas, which are involved with metabolic processes. The precise targeting reduces the likelihood that the estrogen will produce negative effects, such as cancer and stroke.

    Brian Finan, a former doctoral student in DiMarchi's lab, is the lead author of the paper, "Targeted estrogen delivery reverses the metabolic syndrome." Co-authors include Bin Yang and Vasily Gelfanov, research scientists in the IU Bloomington Department of Chemistry, and DiMarchi. Finan is now a post-doctoral researcher at the Helmholtz Zentrum München in Germany, directed by Tschöp, who is DiMarchi's longtime collaborator and a corresponding co-author. Affiliations of the other 20 co-authors include the University of Cincinnati where, also led by Tschöp, many of the in vivo pharmacology and molecular mechanism studies were conducted; Northwestern University; and research laboratories in Germany and China.

    Associated with what health authorities are calling a global epidemic of obesity, the metabolic syndrome consists of obesity associated with other factors such as high blood pressure, high triglycerides, hyperglycemia and low HDL cholesterol. The International Diabetes Federation estimates that as much as 20 percent of the world's adult population has some form of the metabolic syndrome and that they are three times as likely to have a heart attack or stroke and five times as likely to develop adult-onset diabetes as people without the syndrome.

    DiMarchi said investigation continues in the optimization of the peptide-based hormone conjugates with an emphasis on determining the specific mechanism of biological action and identification of an optimal drug candidate suitable for human study. The combination of other peptides and nuclear hormones for targeting other medical conditions holds considerable promise and opportunity for future research.

    Partial funding of the research was provided by Marcadia Biotech and more recently Roche Pharma. Marcadia is a company that DiMarchi co-founded and was acquired in 2010 by Roche Pharma, to which he remains a research consultant.

     

    Published in News
    Monday, 05 August 2013 11:37

    BB50-AU Bullet Blender 50 Gold

    bullet blender homogenizer gentaur antibodiesThe Bullet Blender Gold 50 combines dry ice cooling to keep samples near 4°C, noise insulation to keep homogenization quiet, and extra-powerful homogenization for tough samples. This homogenizer for tissue or cell cultures, can process up to 8 samples in parallel, each sample up to 3.5 g each in a 50 mL tube (Axygen® or Corning® brand 50 mL self standing tube with plug-seal cap).

    Includes the following:

    The Bullet Blender® Gold
    Operator's manual
    Basic starter kit which includes:
    Bead Sample Pack BSP-50B contains 20mL centrifuge tubes of several different types of beads
    Spoon for dispensing beads***

    Cat.No: BB50-AU

    Price: 4985 Euro

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    More form Gentaur: Order Beads for Bullet Blender

    Published in Top Products

    Proteins found in soybeans could inhibit the growth of colon, liver and lung cancers.  Soybean meal is a bi-product following oil extraction from soybean seeds. It is rich in protein, which usually makes up around 40% of the nutritional components of the seeds and dependent on the line, and can also contain high oleic acid (a monounsaturated omega-9 fatty acid).

    The study looked at the role soybeans could have in the prevention of cancer. Using a variety of soybean lines which were high in oleic acid and protein, the researchers looked to monitor bioactivity between the peptides derived from the meals of soybean and various types of human cancer cells.

    The study showed that peptides derived from soybean meal significantly inhibited cell growth by 73% for colon cancer, 70% for liver cancer and 68% for lung cancer cells using human cell lines. This shows that the selected high oleic acid soybean lines could have a potential nutraceutical affect in helping to reduce the growth of several types of cancer cells.

    Published in News

    The polymerase chain reaction (PCR) is a common technique used to amplify, or copy, pieces of DNA. Amplified DNA is then used in genetic analyses for everything from medicine to forensics. In plant research, PCR is a vital step in detecting and sequencing genes, and its applications are endless. However, compounds found in plants often inhibit PCR. Researchers at the University of Southern Mississippi discovered that the use of an additive allows PCR to successfully amplify DNA from once problematic plants.

    PCR is widely used in plant sciences but is not 100 percent reliable. Many plant researchers encounter roadblocks when implementing PCR. For example, many plant species contain phenolic compounds that deter herbivores. These compounds are often extracted along with plant DNA and can stop PCR from working.

    Graduate student Tharangamala Samarakoon and colleagues have found a technique to overcome many of these inhibitory plant compounds. They added a reagent to the PCR mixture that contains three ingredients: trehalose, bovine serum albumin, and polysorbate-20 (all three abbreviated TBT-PAR). "Unlike several other studies, TBT-PAR works at the PCR stage instead of at the DNA extraction stage, so it has promise for pigeon-holed and half-forgotten extractions that previously failed to be amplified using PCR," says Samarakoon. The authors published their research in the January issue of Applications in Plant Sciences.

    Samarakoon tested the TBT-PAR reagent on DNA extracted from tropical and temperate species across four plant families, including Achariaceae, Asteraceae, Lacistemataceae, and Samydaceae. PCR with TBT-PAR successfully amplified DNA for all species, whereas standard DNA extraction and PCR techniques consistently failed.

    TBT-PAR enhanced PCR for DNA extracted from fresh, silica-dried, and herbarium plant material. "Since we study tropical plants, many of which are geographically restricted or rare," explains Samarakoon, "herbarium material is sometimes all that we have available for DNA extraction, and curators are gracious to allow even a small destructive sampling for a single extraction attempt. We want that one attempt, of course, to be successful." Samarakoon predicts that inhibitory plant compounds could be the underlying cause of many PCR failures in herbarium specimens and hopes TBT-PAR will have widespread benefits in herbarium specimen DNA amplification. 

    TBT-PAR was first used in the PCR detection of a shrimp virus by co-author Shiao Wang and his colleagues. "The additive has also been helpful in a colleague's lab where they had trouble amplifying DNA from gopher tortoise ticks, so its utility extends beyond plants," comments Samarakoon. TBT-PAR has the potential for broad use in PCR techniques across DNA samples, species, and taxa.

    The article will be published in the first issue of Applications in Plant Sciences (APPS), a new journal released by the Botanical Society of America. Theresa Culley, Editor-in-Chief of APPS, describes the new journal as a venue to "expedite the dissemination of innovative information encompassing all areas of the plant sciences, including but not limited to genetics, structure, development, evolution, systematics, and ecology."APPS publishes new methods in plant sciences -- an important niche to fill in an age of rapid technological advances.

    Published in News

    Important new research from UMass Medical School demonstrates how exosomes shuttle proteins from neurons to muscle cells where they take part in critical signaling mechanisms, an exciting discovery that means these tiny vehicles could one day be loaded with therapeutic agents, such as RNA interference (RNAi), and directly target disease-carrying cells.

    The study, published this month in the journal Neuron, is the first evidence that exosomes can transfer membrane proteins that play an important role in cell-to-cell signaling in the nervous system.

    "There has been a long-held belief that certain cellular materials, such as integral membrane proteins, are unable to pass from one cell to another, essentially trapping them in the cell where they are made," said Vivian Budnik, PhD, professor of neurobiology and lead author of the study. "What we've shown in this study is that these cellular materials can actually move between different cell types by riding in the membrane of exosomes.

    "What is so exciting about this discovery is that these exosomes can deliver materials from one cell, over a distance, to a very specific and different cell," said Dr. Budnik. "Once inside the recipient cell, the materials contained in the exosome can influence or perform processes in the new cell. This raises the enticing possibility that exosomes can be packed with gene therapies, such as RNAi, and delivered to diseased cells where they could have a therapeutic effect for people."

    Discovered in the mid-80s, exosomes have only recently attracted the attention of scientists at large, according to Budnik. Exosomes are small vesicles containing cellular materials such as microRNA, messenger RNAs (mRNAs) and proteins, packaged inside larger, membrane-bound bodies called multivesicular bodies (MVBs) inside cells. When MVBs containing exosomes fuse with the cell plasma membrane, they release these exosome vesicles into the extracellular space. Once outside the cell, exosomes can then travel to other cells, where they are taken up. The recipient cells can then use the materials contained within exosomes, influencing cellular function and allowing the recipient cell to carry out certain processes that it might not be able to complete otherwise.

    Budnik and colleagues made this startling discovery while investigating how the synapses at the end of neurons and nearby muscle cells communicate in the developing Drosophila fruit fly to form the neuromuscular junction (NMJ). The NMJ is essential for transmitting electrical signals between neurons and muscles, allowing the organism to move and control important physiological processes. Alterations of the NMJ can lead to devastating diseases, such as muscular dystrophy and Amyotrophic lateral sclerosis (ALS). Understanding how the NMJ develops and is maintained is important for human health.

    As organisms develop, the synapse and muscle cell need to grow in concert. If one or the other grows too quickly or not quickly enough, it could have dire consequences for the ability of the organism to move and survive. To coordinate development, signals are sent from the neuron to the muscle cell (anterograde signals) and from the muscle cell to the neuron (retrograde signals). However, the identity of these signals and how their release is coordinated is poorly understood.

    Normally, the vesicle protein Synaptotagmin 4 (Syt4) is found in both the synapse and the muscle cells. Previous knockout experiments eliminating the Syt4 protein from Drosophila have resulted in stunted NMJs. Suspecting that Syt4 played an important role in retrograde signaling at the developing NMJ, Budnik and colleagues used knockdown experiments to decrease Syt4 protein levels in either the neurons or the muscle cells. Surprisingly, when RNAi was used to knockdown Syt4 in the neurons alone, Syt4 protein was eliminated in both neurons and muscles. The opposite was not the case. When Syt4 was knocked down in muscle cells only, there was no change in the levels of Syt4 in either muscles or neurons.

    To confirm this, Budnik and colleagues inserted a Syt4 gene into the neurons of a Drosophila mutant completely lacking the normal protein. This restored Syt4 in both neurons and muscle cells. Further experiments suggested that the only source of Syt4 is the neuron. These observations were consistent with the model that Syt4 is actually transferred from neurons to muscle cells. As a transmembrane protein, however, Syt4 was thought to be unable to move from one cell to another through traditional avenues. How the Syt4 protein was moving from neuron to muscle cell was unclear.

    Knowing that exosomes had been observed to carry transmembrane proteins in other systems and from their own work on the Drosophila NMJ, Budnik and colleagues began testing to see if exosomes could be the vehicle responsible for carrying Syt4 form neurons to muscles. "We had previously observed that it was possible to transfer transmembrane proteins across the NMJ through exosomes, a process also observed in the immune system," said Budnik. "We suspect this was how Syt4 was making its way from the neuron to the muscle."

    When exosomes were purified from cultured cells containing Syt4, they found that exosomes indeed contained Syt4. In addition, when these purified exosomes were applied to cultured muscle cells from fly embryos, these cells were able to take up the purified Syt4 exosomes. Taken together, these findings indicate that Syt4 plays a critical role in the signaling process between synapse and muscle cell that allows for coordinated development of the NMJ. While Syt4 is required to release a retrograde signal from muscle to neuron, a component of this retrograde signal must be supplied from the neuron to the muscle. This establishes a positive feedback loop that ensures coordinated growth of the NMJ. Equally important is the finding that this feedback mechanism is enabled by the use of exosomes, which can shuttle transmembrane proteins across cells.

    "While this discovery greatly enhances our understanding of how the neural muscular junction develops and works, it also has tremendous promise as a potential vector for targeted genetic therapies," said Budnik. "More work needs to be done, but this study significantly supports the possibility that exosomes could be loaded with therapeutic agents and delivered to specific cells in patients."

    Published in News
    Tuesday, 30 July 2013 12:54

    Herc1-set siRNA/shRNA/RNAi Lentivector

    Species Mouse
    Accession Number NM_145617.3
    Vector piLenti-siRNA-GFP
    Target Sequence Sequence available upon placing order
    Sequence Primers Sequence available upon placing order
    User Manual  
    Bacterial Selection Kanamycin
    Mammalian Selection Puromycin
    Format Plasmid
    Appearance Liquid
    Storage -20°C or below
    Shelf Life 1 year (when at -20°C or below in a non-frost free freezer)
    Shipping Shipped at ambient temperature
    Quality Control Restriction Enzyme Digest and Sequencing
    Caution This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information.
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       Our RNA interference lentiviral vectors contain siRNAs. We employ a dual convergent promoter system where the sense and antisense strands of the siRNA are expressed by two different promoters rather than in a hairpin loop - to avoid any possible recombination events that can occur. 
       Gentaur guarantees that at least one out of the four siRNA Lentivector constructs purchased in a set will result in over 70% knockdown of gene expression within target cells showing >80% transfection efficiency. If this is not the case, we will provide a one-time replacement of four new constructs with alternative siRNA sequences. To qualify for this replacement, the vectors must be transfected at ≥5 nM and assayed 48 hours post-transfection. Customers must provide adequate data to show >80% transfection efficiency with a positive control, plus additional qPCR data or a western blot to evaluate the level of gene expression. The replacement set will not covered by the same guarantee, and if these constructs are also considered to be ineffective then it is most likely the gene is not susceptible to siRNA knockdown.
       Gentaur limits its obligation and liability for the success of this technology to providing one replacement of any siRNA lentivector product only. Before sending your inquiry, please make sure you have optimized your experiments as far as possible, this includes (where applicable) increasing your MOI and the duration of infection (up to 72 hours), and carrying out clone screening before assaying for knockdown. 

    Published in Promos

    hiv treatment gentaur antibodiesUntreated HIV infection destroys a person's immune system by killing infection-fighting cells, but precisely when and how HIV wreaks this destruction has been a mystery until now. New research by scientists at the National Institute of Allergy and Infectious Diseases, part of the National Institutes of Health, reveals how HIV triggers a signal telling an infected immune cell to die. This finding has implications for preserving the immune systems of HIV-infected individuals.

    HIV replicates inside infection-fighting human immune cells called CD4+ T cells through complex processes that include inserting its genes into cellular DNA. The scientists discovered that during this integration step, a cellular enzyme called DNA-dependent protein kinase (DNA-PK) becomes activated. DNA-PK normally coordinates the repair of simultaneous breaks in both strands of molecules that comprise DNA. As HIV integrates its genes into cellular DNA, single-stranded breaks occur where viral and cellular DNA meet. Nevertheless, the scientists discovered, the DNA breaks during HIV integration surprisingly activate DNA-PK, which then performs an unusually destructive role: eliciting a signal that causes the CD4+ T cell to die. The cells that succumb to this death signal are the very ones mobilized to fight the infection.

    According to the scientists, these new findings suggest that treating HIV-infected individuals with drugs that block early steps of viral replication -- up to and including activation of DNA-PK and integration -- not only can prevent viral replication, but also may improve CD4+ T cell survival and immune function. The findings also may shed light on how reservoirs of resting HIV-infected cells develop and may aid efforts to eliminate these sites of persistent infection.

    Published in News

    immune cell gentaur antibodiesMelbourne researchers have discovered that the death of immune system cells is an important safeguard against the development of diseases such as type 1 diabetes, rheumatoid arthritis and lupus, which occur when the immune system attacks the body's own tissues.

    The finding suggests that these so-called autoimmune diseases could be treated with existing medications that force long-lived immune system cells to die.

    In the development of the immune system, some cells are produced that have the potential to attack the body's own tissues, causing autoimmune disease. The death of these 'self-reactive' immune cells through a process called apoptosis is an important safeguard against autoimmune disease.

    But Dr Kylie Mason, Dr Lorraine O'Reilly, Dr Daniel Gray, Professor Andreas Strasser and Professor David Huang from the Walter and Eliza Hall Institute, and Professor Paul Waring from the University of Melbourne have discovered that when immune cells lack two related proteins, called Bax and Bak, the cells can attack many healthy tissues, causing severe autoimmune disease. The research is published online January 22 in the journal Proceedings of the National Academy of Sciences.

    Bax and Bak are members of the 'Bcl-2 protein family', a large group of proteins that control cell death. Dr O'Reilly said it was thought that Bax and Bak acted like an irreversible switch in cells, determining when cells die by apoptosis. In healthy cells, Bax and Bak are in an 'inactive' form, but when cells are under stress or receive external signals instructing them to die, Bax and Bak switch into an 'active' form that starts the destruction of the cell by apoptosis. Without Bax and Bak, cells are highly protected against apoptosis.

    Dr O'Reilly said that some immune cells that lacked the proteins Bax and Bak were able to attack healthy tissues in many organs of the body. "Normally, these 'self-reactive' immune cells are deleted during development," she said. "In the absence of Bax and Bak, enough self-reactive cells survive development to persist in the body and cause autoimmune disease in organs such as the kidneys (glomerulonephritis), similar to what is seen in the most severe form of lupus.

    "Our findings confirm that defective apoptosis of immune cells can cause autoimmune disease, and that Bax and Bak are important determinants of immune cell death. We were also interested to see that, in our model, loss of Bak on its own was sufficient to cause autoimmune disease, albeit to a lesser extent than losing both Bak and Bax. This supports a recent discovery that humans with mutations in the BAK gene are predisposed to certain autoimmune diseases."

    The research provides hope for people with autoimmune diseases as Bax and Bak activity can be triggered by a new class of potential anti-cancer agents, called BH3-mimetics, which are currently in clinical trials for certain types of leukemia in Melbourne, Dr O'Reilly said. "Our findings suggest that BH3-mimetics might be an exciting new option for treatment for autoimmune conditions, by activating Bax and Bak and making the self-reactive immune cells which are causing the autoimmune disease to die," she said.

    Published in News

    Methadone reduces the risk of HIV transmission in people who inject drugs (PWID), as reported by an international team of researchers in a paper published October 5 in the online edition of theBritish Medical Journal. This team included Dr. Julie Bruneau from the CHUM Research Centre (CRCHUM) and the Department of Family Medicine at the University of Montreal.

    "There is good evidence to suggest that opiate substitution therapies (OST) reduce drug-related mortality, morbidity and some of the injection risk behaviors among PWID. However, to date there has been no quantitative estimate of the effect of OST in relation to HIV transmission. This new study provides solid evidence demonstrating the link between these treatments and a reduced risk of HIV transmission," notes Dr. Bruneau, one of the six investigators who worked with Dr. Matthew Hickman, the study's principal investigator and Professor in Public Health and Epidemiology at the University of Bristol (UK).

    "These results are important given that increases in HIV incidence have been reported among PWID in a number of countries in recent years, where opiate substitution therapies are illegal or severely restricted," adds Dr. Bruneau. 

    Injection drug use is a major risk factor for the transmission of HIV and AIDS. It is estimated that around 5-10% of HIV infections worldwide are due to injection drug use. Methadone and buprenorphine are the main forms of drug prescribed for addicts and are frequently prescribed as opiate substitution therapies.

    The results of this study are the fruit of an international collaborative effort. Authors from the US, Canada, Italy and Australia carried out a review and pooled analysis (known as a meta-analysis) of several published and unpublished studies from multiple countries (including the USA, Canada, the UK, the Netherlands, Austria, Italy, Thailand, Puerto Rico and China) to determine the association between OST and HIV transmission among PWID. The nine selected studies looked predominantly at males between 26 and 39 years old and totalled 819 incidents of HIV infection with 23,608 person-years of follow-up.

    After analysing these studies, authors found that OST was associated with a 54% reduction in risk of HIV infection among PWID. There were differences between the studies, including different background rates of HIV infection, making it impossible to calculate an "absolute risk reduction" for HIV infection that would translate to all settings. And not all studies reported adjustments to the intervention to take account of other factors that might influence the association between OST and HIV infection. But the impact of OST on HIV was strong and consistent in further analyses in the paper. There was weak evidence to suggest that longer duration of OST exposure may be associated with greater benefit.

    For Dr. Bruneau, the results of this study favour the promotion of opiate substitution therapies: "These therapies can reduce HIV transmission among PWID not only in countries in which there is a high incidence of this disease, but also in Quebec where there has been an increase in the use of illicit opiates intravenously, particularly among youths, and where access to OST is problematic."

    Published in News
    Thursday, 25 July 2013 10:50

    Automatic Biochemistry Analyzer 9050

    biochemistry analyzer gentaur 9050Specification

    Throughput: 400 tests/hour, 800 test with ISE

    Measuring wavelength: 340 to 800, 12 wavelengths invariable.

    Assay tapes: End point, Kinetics, Fixed time, Immunoturbidimetry, Monochromatic, Bi-chromatic & Multi-standard Analysis, Linearity & Nonlinearity, Calibration, Single & Dual Reagents can be used

    Calibration Mode: Linear (one-point, two-point and multi-point), Logit-log4p, logit-log5p Spine, Exponential, Polynomial, Parabola.

    Light source: Halogen lamp 6v/10w, life span ≥3000 hours

    Cuvette: light pass length: 6mm

    Reaction disk: Turntable

    Reaction temperature: 37℃± 0.1 ℃

    Sample Volume: 2 to 35μL (in steps of 0.1μL)

    Power supply: 220V AC,50/60 Hz

    Price: 23 500 Euro

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    More: Biochemistry Analyzer

    Published in Promos