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Thank you B.Dhungel, from Gallipoli Medical Research Foundation (GMRF), Greenslopes Private Hospital, University of Queensland for such a great review.
I am really impressed by the reproduciblity of the Bioline Isolate II RNA minikit, the cDNA sysnthesis and the Sybr-Lo Rox mix in qPCR. We consistently get high concentrations of pure RNA with 260/280 reading 2 when measured with the nanodrop.
We have seen that the cDNA synthesis kit from Bioline is able to synthesize high quality cDNA even from low starting RNA concentrations when analyzing the qPCR results.
Are you a scientist at the leading edge of biological discovery? Then join Bioline at the Analytica International Trade Fair in Munich, April 1-4, 2014.
Visit us on stand 516 in Hall A3 to explore our research solutions for gene expression and genotyping studies. Discover how our innovative products help you to achieve better and more accurate results.
This year we’ll be showcasing our new products SensiFAST™ cDNA Synthesis Kit, the unmatched MyTaq™ Blood PCR Kit, and the latest addition to our range, EPIK™ Kits for Epigenetics research and analysis.
Four live talks a day at Analytica 2014
We'll be hosting live talks on the stand on all four days of the event, covering gene expression and cDNA synthesis, high-sensitivity qPCR, Bisulfite PCR, and all the latest innovations in Direct PCR.
Attend one of our talks most suited to your research, find out why Bioline is the perfect partner for your research, AND receive a free Bioline T-shirt!
We very much look forward to meeting with you at Analytica 2014!
The World Health Organisation (WHO) and the Centre for Disease Control & Prevention (CDC) are working together to better understand the threats posed by two novel RNA viruses causing human infections and deaths. In Saudi Arabia, 30 confirmed cases of the novel coronavirus (nCoV) have been reported to date, including at the time of writing fifteen deaths. This coronavirus is different from any other coronaviruses that have previously been found in people. It is related to the coronavirus responsible for causing Severe Acute Respiratory Syndrome (SARS), which claimed the lives of 648 people over a decade ago in China and Hong Kong.
The shadow of SARS still hovers over the region since the emergence of the novel avian influenza A H7N9 virus. The WHO has called the H7N9 strain "one of the most lethal" flu viruses ever seen. To date, 31 people have died and 130 people have been diagnosed with H7N9. Many of the people infected with H7N9 are reported to have had contact with poultry. However some cases reportedly have not had contact with poultry. Sustained human-to-human transmission has, fortunately, not yet been observed. However, if the virus were to adapt and spread readily between people, it will pose a much greater threat and scientists have already warned that the virus is mutating rapidly.
The nCoV and H7N9 viruses share some similarities in that neither have been diagnosed in humans before. People of all ages have little protective immunity and a global pandemic could be a possibility. Both viruses can lead to severe disease, characterised by high fever, severe respiratory illness and, potentially, death.
Furthermore, the origins of both viruses have yet to be firmly established. A new study in the Lancet suggests H7N9 originated from chicken and duck influenza viruses.
"The novel avian influenza A H7N9 virus might have evolved from at least four origins," Professor Gao from the Chinese Center for Disease Control and Prevention in Beijing and his co-authors concluded. "Unknown intermediate hosts involved might be implicated, extensive global surveillance is needed, and domestic-poultry-to-person transmission should be closely watched in the future."
China has been working on developing vaccines and other treatments for H7N9 and has slaughtered thousands of birds and closed many poultry markets in the hope of slowing the outbreak. On Monday, the CDC sought to assure the public of H7N9's relative limitations, saying the current strain is not capable of delivering a global pandemic.
"This particular virus is not going to cause a pandemic because it doesn't spread person-to-person," Dr. Thomas Frieden, director of the CDC, told Reuters. "But all it takes is a bit of mutation for it to be able to go person-to-person. I cannot say with certainty whether that will happen tomorrow, within 10 years or never."
Experimenting with H5N1: controversial science
A controversial new Science paper showing genetic re-assortment experiments with the avian H5N1 influenza A virus has also hit the headlines. Researchers from the Harbin Veterinary Research Institute led by Prof. Chen, director of China's National Avian Influenza Reference Laboratory mixed the H5N1 avian-flu virus, which is highly lethal but not easily spread between people, with a 2009 strain of H1N1 flu virus, which is very infectious to humans. The experiments resulted in 127 different viral hybrids between H5N1 and H1N1. Five H5N1-H1N1 hybrids ‘gained' the ability to infect guinea pigs, via airborne transmission.
Professor Chen commented that “The studies demonstrated that H5N1 viruses have the potential to acquire mammalian transmissibility by re-assortment with the human influenza viruses. This tells us that high attention should be paid to monitor the emergence of such mammalian-transmissible virus in nature to prevent a possible pandemic caused by H5N1 virus,” she said. It is difficult to say how easy this will happen, but since the H5N1 and 2009/H1N1 viruses are widely existing in nature, they may have a chance to re-assort,” she added.
In January, a global moratorium on certain controversial experiments involving H5N1 was lifted. This was sparked by papers in 2011 by two research groups from Japan and Holland who demonstrated how the virus, which normally infects birds, could gain the ability to move between mammals via mutations in a single gene – hemagluttinin, a viral surface protein. The debate over whether the results should have been published has been extremely heated. Critics have raised concerns that the data could be used by terrorists, should they happen to acquire the technology, to unleash a pandemic and have also attacked the decision to lift the moratorium.
Bioline offers SensiFAST™, a range of rapid and highly sensitive real-time PCR kits for the detection and study of viruses in relation to public health. For RNA viruses such as Influenza and Coronavirus, Bioline offers the new generation of advanced formulation SensiFAST™ One-Step Probe Kits. The 2x mastermix offers superior reproducibility and sensitivity for quantification of viral RNA genetic sequences in a convenient and easy-to-use format.
For further information, please see:
Focus on Breast Cancer
For years scientists have been treating breast cancer as a single disease. However, a new landmark study published in Nature has reclassified breast cancer into ten separate sub-diseases based on their genetic fingerprint. The culmination of decades of research, the study is the largest global study of breast cancer tissue ever performed.
The team, led by the British Columbia Cancer Center in Canada and the Cambridge Cancer Research Institute in the UK, used genome-wide microarrays to analyze the DNA and RNA of 2,000 tumor samples taken from women diagnosed with breast cancer. This huge pool of genetic information (copy number variants, SNPs and gene expression data), as well as survival data, allowed researchers to spot new and previously unacknowledged patterns for ten subtly different cancers that have, historically, been considered as one.
The challenge now is to understand the genetic drivers behind these newly discovered breast cancer variants and to develop new targeted therapies in the future. It could also lead to women with the best prognosis being spared side-effects of chemotherapy. The classification system will likely also form the basis for newer and better ways to diagnose and manage the disease.
Bioline offers a number of reagents that have helped further the study of cancers and, more specifically, breast cancer. So this edition of Bioline Scholar Monthly focuses on the use of Bioline reagents and kits in the field of breast cancer research.
In a diverse cohort of breast cancer patients with a 1–5 year tumor relapse versus those with up to 7 years relapse-free survival, RNA was extracted and subjected to microarray and real-time RT-PCR analysis. Among the 299 genes, five genes which included B cell response genes were found to predict with >85% accuracy relapse-free survival. Real-time RT-PCR confirmed the 5-gene prognostic signature that was distinct from an FDA-cleared 70-gene signature of MammaPrint panel and from the Oncotype DX recurrence score assay panel.
Ascierto, L. M., et al. Breast Can. Res. Treat. 131(3):871-880 (2012) - A signature of immune function genes associated with recurrence-free survival in breast cancer patients.
MicroRNAs (miRNAs) are noncoding RNAs that function as key posttranscriptional regulators of gene expression. This paper found that BRCA1 recognizes the RNA secondary structure and directly binds with primary transcripts of miRNAs via a DNA-binding domain. The findings indicate novel functions of BRCA1 in miRNA biogenesis, which may be linked to its tumor suppressor mechanism and maintenance of genomic stability.
Kawai S. and Amano A. J. Cell Biol. 197 (2):201-208 (2012) - BRCA1 regulates microRNA biogenesis via the DROSHA microprocessor complex.
The bioactive lipid sphingosine 1-phosphate (S1P) uses sphingosine 1-phosphate receptor 4 (S1P4) and human epidermal growth factor receptor 2 (HER2) to stimulate the extracellular signal regulated protein kinase 1/2 (ERK-1/2) pathway in MDA-MB-453 cells. The magnitude of the signaling gain on the ERK-1/2 pathway produced in response to S1P can be increased by HER2 in MDA-MB-453 cells. The linkage of S1P with an oncogene suggests that S1P and specifically S1P4 may have an important role in breast cancer progression.
Long J. S., et al. J. Biol. Chem. 285:35957-35966 (2010) - Sphingosine 1-Phosphate Receptor 4 Uses HER2 (ERBB2) to Regulate Extracellular Signal Regulated Kinase-1/2 in MDA-MB-453 Breast Cancer Cells.
CD44, the transmembrane receptor for hyaluronan, is implicated in tumor cell invasion and metastasis. The expression of CD44 and its variants is associated with poor prognosis in breast cancer. This paper investigated the effect of silibinin (a polyphenolic flavonolignan of the herbal plant of Silybum marianum, milk thistle) on the epidermal growth factor (EGF) ligand-induced CD44 expression in human breast cancer cells. The results suggest that silibinin prevents the EGFR signaling pathway and may be used as an effective drug for the inhibition of metastasis of human breast cancer.
Kim S., et al. Anticancer Res. 31(11): 3767-3773 (2011) - Silibinin Suppresses EGFR Ligand-induced CD44 Expression through Inhibition of EGFR Activity in Breast Cancer Cells.
Human papillomavirus (HPV) and Epstein Barr virus (EBV) have been found in breast carcinomas around the world. In this study, fifty-five BCs from Chile were analyzed for HPV and EBV presence. In addition, HPV- 16 viral load/physical status and E6/E7 expressions were determined. The results suggest that it is unlikely that HPV and/or EBV play a direct role in the etiology of breast carcinomas.
Aguayo F., et al. Infectious Agents and Cancer 6:7 (2011) - Human papillomavirus and Epstein-Barr virus infections in breast cancer from chile.
This study suggests that melatonin may play a role in the desmoplastic reaction in breast cancer through a down regulatory action on the expression of antiadipogenic cytokines, which decrease the levels of these cytokines. Lower levels of cytokines stimulate the differentiation of fibroblasts and decrease both aromatase activity and expression, thereby reducing the number of estrogen-producing cells proximal to malignant cells.
Alonso-González C., et al. J. Pineal Res. 52(3): 282–290, (2012) - Melatonin interferes in the desmoplastic reaction in breast cancer by regulating cytokine production.
Melatonin reduces the development of breast cancer interfering with oestrogen-signalling pathways, and also inhibits aromatase activity and expression. This study shows that melatonin inhibits aromatase activity and expression by regulating the gene expression of specific aromatase promoter regions. A possible mechanism for these effects would be the regulation by melatonin of intracellular cAMP levels, mediated by an inhibition of cyclooxygenase activity and expression.
Martínez-Campa C., et al. British J. Can. 101: 1613–1619 (2009) - Melatonin inhibits aromatase promoter expression by regulating cyclooxygenases expression and activity in breast cancer cells.
Bisphenol A (BPA) has long been suspected to promote carcinogenesis, but the high doses of BPA used in many studies generated conflicting results. This paper shows that BPA at environmentally relevant doses reduces the efficacy of chemotherapeutic agents. These data provide considerable support to the accumulating evidence that BPA is hazardous to human health.
LaPensee E. W., et al. Environ Health Perspect. 117(2): 175–180 (2009) - Bisphenol A at Low Nanomolar Doses Confers Chemoresistance in Estrogen Receptor-a–Positive and –Negative Breast Cancer Cells.
This paper indicates that decreased CDH1, CDH13 and TIMP3 with increased CD44 gene expression levels can be used as an indicator for invasion in both ER-positive and ER-negative breast tumors. In double-negative tumor tissues, CD44 can be considered a marker for aggressive properties. However, additional assays in a larger series of patients with long follow up will be necessary to confirm these results of gene expressions in ER-positive and ER-negative tumors and their relationship with HER2 and ESR1.
Celebiler A., et al. Can. Sci. 100: 2341–2345. (2009) - Predicting invasive phenotype with CDH1, CDH13, CD44, and TIMP3 gene expression in primary breast cancer.