rilpoint_mw113

VATIS Update Biotechnology . May-Jun 2005

Register FREE
for additional services
Biotechnology May-Jun 2007

ISSN: 0971-5622

VATIS Update Biotechnology is published 4 times a year to keep the readers up to date of most of the relevant and latest technological developments and events in the field of Biotechnology. The Update is tailored to policy-makers, industries and technology transfer intermediaries.

Co-publisher: Biotech Consortium India Ltd
Editorial Board
Latest Issues
New and Renewable
Energy
VATIS Update Non-conventional Energy Jul-Sep 2017
Bio
Technology
VATIS Update Biotechnology Jul-Sep 2017
Waste
Management
VATIS Update Waste Management Oct-Dec 2016
Food
Processing
VATIS Update Food Processing Oct-Dec 2016
Ozone Layer
Protection
VATIS Update Ozone Layer Protection Sep-Oct 2016
Tech
Monitor
Asia-Pacific Tech Monitor Oct-Dec 2014

Contents

__NOTOCNUM__

IN THE NEWS

GM plants to grow vaccines against killer diseases

With a grant of 12 million from the Sixth Framework Programme (FP6) of the European Union, genetically modified (GM) plants are to be used to grow vaccines for use in the worldwide fight against HIV, tuberculosis, diabetes and rabies. Because plants are inexpensive to grow, they could be used to produce large quantities of drugs or vaccines at low cost from 10 to 100 times lower than conventional production, which is often labour-intensive, expensive and produces relatively small amounts of pharmaceuticals.


The programme known as Pharma-Planta draw on the expertise and experience of 39 scientists from 11 European countries and South Africa to address major health problems affecting Europe and the developing world. In the first international project of its kind, Pharma-Planta would develop the concept from plant modification through to clinical trials, and human trials are expected within four years. On the project being successful, the techniques would be licensed to the developing countries. They would then be able to start up their own production to generate whatever amount they require at a cost that would not impact greatly on the countries economy. Although the project has not finally decided which plants would be used, the likely candidates are tobacco or maize.


Genetic modification of other organisms is already being used to produce human insulin and a hepatitis B vaccine. The use of plant-derived materials in humans has never been formally addressed within the European Union. Pharma-Planta is therefore a ground-breaking project, and aims to provide help for the millions of people that die each year throughout the world from vaccine-preventable diseases.


Website: www.agbios.com

China ratifies Cartagena Protocol on Biosafety

The State Environmental Protection Administration (SEPA) of China has announced the countrys ratification of the Cartagena Protocol on Biosafety, which regulates the international trade of genetically modified organisms (GMO). According to Dr. Wan Bentai, Director of SEPAs Natural Ecosystem department, the ratification of the protocol demonstrated Chinas commitment to enhancing management on biosafety in line with international law, which promotes the development of legislation on the subject, strengthens the countrys management of GMO cross-border movement, improves GMO labelling systems and promotes the publics involvement in biosafety.


The Cartagena Protocol obligates exporters to give greater information to recipient nations about GMO products. It also gives importers the power to reject GMO imports or donations, even without scientific proof, if believed that these pose a danger to traditional crops and indigenous societies. The Protocol, a part of the Convention on Biological Diversity, came into effect in September 2003 and has since been ratified by 120 countries.


Although China has made much progress in GMO research since the 1980s, there were still many loopholes in its management of the biosafety sector such as ineffective supervision in the face of growing import of GMO products. Ratification of the protocol was also part of the countrys efforts to conserve its biodiversity, which is under serious threat.


Website: www.agbios.com

Biotech parks in India nearing completion

The Karnataka Biotech Park, known as Bangalore Helix, in Bangalore, India, is fast nearing completion. The Department of Biotechnology, Government of India, has sanctioned US$3 million for the project, the estimated total value of which is US$5.5 million. The state governments allocation of US$1.44 million in the state budget was later increased to US$2.55 million. The biotech park will have world-class infrastructure on 100 acres of land, which is already in the possession of the Department of IT and Biotechnology. A number of biotech companies have shown interest to set up their operation in the park.


The first phase of the Kinfra Biotechnology Park, Kalamassery, Kerala, will be completed by August 2005. At an investment of US$5.15 million, the park will have an advanced Technology Incubation Centre (TIC) and a general instrumentation laboratory. The park is a joint venture of the state governments infrastructure development arm, the Kerala Industrial Infrastructure Development Corporation (Kinfra), and the Department of Biotechnology (DBT) of the Government of India. The park spanning an area of 50 acres is coming up in a 240-acre plot owned by Kinfra.


The first phase covers biotech R&D facilities in an area of 10,000 sq. ft space, set up as 10 modules of 1,000 sq. ft each, and a common instrumentation lab. DBT, which has funded US$2.44 million to Kinfra for the park, is giving guidance for setting up the TIC.


Chronicle Pharmabiz, 15 May 2005, 28 April 2005

Kenyan scientists trial plant pest-proof GMO maize

A research team led by Dr. Simon Gichuki at the Kenya Agriculture Research Institute has begun the trial planting of maize genetically modified to resist insect pests. This is a step towards reducing 20 per cent of crop lost every year because of pests. The scientists hope the variety would resist the stem borer larvae, which is blamed for Kenyas dependence on maize imports, hitting the incomes of farmers and reducing food security.


Kenya produced 2,142,000 tonnes of the staple in 2004 and imported 241,800 tonnes at a cost of about US$60 million. About 400,000 tonnes of maize is lost to pests, especially stem borers like the maize stock borer. This loss directly affects the livelihoods of thousands of families dependent on maize as a staple food and as a source of income. Kenya does not permit genetically modified organism (GMO) foods, but is in the final stages of drafting legislation to govern the use of GMO technology. In addition to maize, there is work going on into possible use of GMO cotton and tuber crops.


Website: www.agbios.com

Philippines clears planting of second biotech corn

The Philippines, which is the first Asian country to commercialize genetically modified corn, has approved the sale and planting of another biotech corn, an insect-resistant Bt-11 strain developed by the Swiss agrochemicals maker Syngenta. Farmers could increase their yields by 40 per cent per harvest by using this corn strain, which contains the genes from Bacillus thuringiensis and is resistant to the Asiatic corn borer pest. The Asiatic corn borer is the leading destroyer of corn crops in the Philippines, causing losses of up to 80 per cent in yields.


According to Mr. Rod Bioco, President of the Philippine Maize Federation Inc., farmers who used Bt corn seeds increased yields by 25-33 per cent to 5-6 tonnes per hectare per crop last year versus the 4-4.5 tonnes using other varieties of yellow corn. However, at US$83-92 per a bag of 50 kg, Bt corn seeds would cost twice as much as hybrid seeds developed in conventional ways. It is hoped that more competition in the market would make the price of corn seeds drop. Officials from the state regulator Bureau of Plant Industry report that the Bt corn has been accepted in the domestic market. Some 52,000 hectares, or two per cent of the total 2.5 million hectares planted corn in the country, were planted with Bt corn in 2004.


Website: www.agbios.com

Small business innovation scheme proposed in India

The Department of Biotechnology (DBT), India, has proposed to institute Small Business Innovation Research Initiative (SBIRI) scheme in 2005-06 for supporting small and medium size enterprises with a grant or loan to help early phase of product development. Those companies with up to 1,000 employees would be able to avail this scheme and preference would be given to proposals that address important national needs.


SBIRI scheme would operate in two phases of innovation and product development. Phase 1 would provide for highly innovative, early-stage, pre-proof of concept research. The maximum amount of funding to an enterprise would be limited to Rs 5 million, with not more than 50 per cent of it going as grant and the remaining as an interest-free loan. For a companys project to be considered at this stage, a partnership with a public R&D institution will not be mandatory for those companies that have good quality scientists, though it would be considered an important factor. This should encourage high quality scientists to work in a small and medium size biotech companies. The proposal is to provide soft loans for product developments and the commercialization at an interest rate of 2 per cent. The role of public R&D institutions at this stage is critical. The partner in the public institution will get the R&D support as grant. Small and medium knowledge-based industries in the biotech sector would also be encouraged to avail of equity support from the SME growth fund of Small Industries Development Bank of India.


Chronicle Pharmabiz, 21 April 2005

Pakistan notifies biosafety rules

The Government of Pakistan has notified biosafety rules to have vigilance on the movement of harmful genetically modified organisms (GMOs) and benefit from useful organisms in agricultural production. These rules would apply to all GMOs like agricultural produces, seeds, plants, crops and animals, and allow their import after risk assessment according to global standards. According to the notification, the government will constitute a National Biosafety


Committee, a Technical Advisory Committee and Institutional Biosafety Committees. Institutions and organizations, engaged in biotechnology or genetic manipulation work, will appoint or designate Biosafety Officers who would assist and liaison with Institutional Biosafety Committees to review regularly the operating procedures and biosafety records, and to assess the integrity of containment facilities and safety equipment/utilities.


Commenting on the biosafety rules, Major (Retd.) Tahir Iqbal, Pakistans Minister for Environment, hoped it would usher in a new era of progress in the agricultural sector. The rules provide that no person shall import, export, sell, purchase or trade living modified organisms, substances or cells and products without obtaining licence from the federal agency (Pakistan Environmental Protection Agency) through an application prepared in conformity with the biosafety guidelines. Under these rules, any person to whom a licence would be granted shall notify the federal agency and the National Biosafety Committee of any change or addition to the information already submitted. The information termed by the applicant as confidential shall not be disclosed. The Licence granted by the federal agency shall not take effect until the applicant executes an undertaking and assumes the legal duty to comply with applicable provisions of biosafety guidelines.


Website: www.agbios.com

Malaysia to launch US$26 million biotech fund

The Government of Malaysia has set up a M$100 million (US$26 million) fund to support biotechnology research and development. The Malaysian Biotechnology Corporation, set up in April 2005 to support and advise biotechnology-related businesses, would distribute part of the money. Earlier, Prime Minister Mr. Abdullah Ahmad Badawi unveiled the countrys national biotechnology policy and highlighted the potential for biotechnology to improve health, increase food security and protect the environment. He said Malaysia was noted for its skilled workforce, strength in research-based industry and excellent infrastructure. But to take advantage of these strengths, the country had to create the right environment for biotechnology to flourish, and assist those with the expertise to carry out biotech-driven research and development or to develop biotech-driven businesses.


Website: www.scidev.com

Genetically modified rice proves beneficial in China

A study of genetically modified rice trials in China, co-authored by University of California, Davis, the United States, and Dr. Scott Rozelle, an agricultural economist, has shown that rural farmers obtained significant health and economic benefits from working with GM rice as compared with non-genetically modified rice growers. The latter were found to buy and use 8-10 times as much pesticide on their crops and subsequently experience adverse health effects.


The study was conducted on the two varieties of GM rice one a Bt rice and the other a GM rice, which was altered with a pest resistant gene from cowpea plant. The two-year study followed the crop yield, pesticides use and health of the GM rice growers and the regular rice growers. GM rice growers reported 6-9 per cent more yield and an 80 per cent reduction in pesticide use compared with farmers using non-GM crops. None of the farmers growing the GM rice reported harmful health effects while, in contrast, up to 10 per cent of non-GM rice growers reported a pesticide-induced illness in a year. The potential benefits of GM rice to China were significant. The Chinese National Bureau of Statistics estimates that 50 per cent of the Chinese population works on farms and millions of farmers stand to gain in terms of health and economic advancement by pesticide reduction. The environment also stands to benefit, as China uses more pesticides than any other country, and the largest share is used on rice.


Website: www.californiaaggie.com

Proposal for diagnostic biotechnology cell in India

The Department of Biotechnology (DBT), India, has proposed a separate cell for diagnostic biotechnology to encourage and support studies into the clinical application of pharmacogenomics. It would encourage research, involving investigators with both clinical practice and pharmacology or pharmacokinetics expertise. Providing incentive for this group of clinician-researchers to bring scientific advances to the patients bedside has been one of the major concerns of DBT in recent times. Encouragement from biopharmaceutical companies to include pharmacogenomics data in their drug submission is another measure that has been taken up at the individual level. DBT has identified that advances in biosensors and gene amplification are in the offing to enable real-time medicine.


Chronicle Pharmabiz, 26 May 2005

South Africa to start GM cotton trials

Field trials are to be conducted on new varieties of super resistant and fast-growing GM cotton in South Africa. According to Dr. Magda du Toit, Monsanto spokesperson, the field trials are part of a larger programme to develop commercial-grade cotton resistant to bollworms and tolerant to herbicides.


Research and seed production trials would cover between 0.3 to 30 hectares at each test site, and the fields would all be surrounded by non-GM cotton to prevent the GM plants from spreading. The fields will be thoroughly harvested to prevent the plants regrowing and seeds resulting from the trials would be shipped to the United States. GM crops have been widely accepted in the United States, and the European Union has also recently released a report that declares GM foods safe, after examining 81 different GM trials over a 15-year period.


Website: www.news24.com

MARKET NEWS

Agilent to bring its genomics technology to India

Agilent Technologies Inc., California, the United States, has announced plans to introduce its DNA microarray-based genomics solutions into India. Agilent expects to expand its presence into industries such as medical research, drug discovery and agro-biotech. Its Life Sciences and Chemical Analysis (LSCA) business had double-digit growth in India in 2004, driven by the countrys booming biotechnology, agriculture, information technology and pharmaceutical industries.


According to Mr. Sanjeev Dhar, LSCA Country Manager, India is now LSCAs fourth largest country of business in Asia and one of the fastest growing into areas such as genomics and proteomics research. Agilents initial plans focus on introducing its DNA microarray-based genomics solutions, which improve the productivity of gene expression and genomic research. Key applications of its microarray-based solutions include gene expression, toxicogenomics, agro-biotech, comparative genomic hybridization, etc.


Chemical Weekly, 10 May 2005

IFC commits US$4 million in APIDC biotech fund

The International Finance Corporation (IFC) the private sector arm of the World Bank Group based in Washington, the United States, is providing equity support of up to US$4 million to APIDC biotechnology fund, a private equity fund formed to invest in start-up and early-stage life sciences business in India. APIDC biotech fund is the first fund in India to focus exclusively on the life sciences sector and seek early-stage opportunities in areas such as healthcare, drug discovery, agriculture, dairy, environmental and industrial applications throughout India.


According to Mr. Sarath Naru, General Partner, APIDC Venture Capital Fund, IFC has played a key role in mobilizing investors, structuring the project, and incorporating high environmental, social, ethical and governance standards. Mr. Iyad Malas, IFCs South Asia Director, hoped that several of the 20-25 companies that the fund would seek to support also want to partner with IFC for later-stage funding.


Chronicle Pharmabiz, 26 May 2005

Bioinformatics market reported as explosive

The global bioinformatics market is currently estimated at about US$1.4 billion and expected to grow at an average annual growth rate of 15.8 per cent to reach nearly US$3 billion by 2010, according to the report RB-206 Bioinformatics Business: Technical Status and Market Prospects. The fastest growing market segment in bioinformatics industry would be analysis software and services, primarily driven by the need for improved and sophisticated tools for analysing and using biological data for developing therapeutic drugs.


The application of bioinformatics in drug discovery and development is expected to reduce the annual cost of developing a new drug by 33 per cent, and the time taken for drug discovery by 30 per cent.


Pharmaceutical companies are expected to increase their R&D expenditure in the future and a major portion of R&D spending is expected to go into bioinformatics. Global spending on drug discovery is estimated to increase to US$25.1 billion in 2006 from US$19.6 billion in 2002. The contribution of genomics in the drug discovery and development process was anticipated to be the highest in the near future, as most of the new drug design in the long term was expected to be genomics-related. However, after human genome mapping, the application of proteomics is tipped to increase at a higher rate than that of genomics, as it is proteins, not genes, which will be targeted by future research for developing new drugs.


Scientists are acquiring genomics data through techniques such as amplification, DNA microarray expression, real-time PCR and genotyping.


Instrumentation, hardware and software are then needed to analyse, integrate and transmit this vast amount of data, which has resulted in major IT challenges for those in the field. The segment is estimated to grow at an average annual growth rate of 21.2 per cent, from US$444.7 million in 2005 to US$ 1.16 billion in 2010. Growth in the genomics-based content would drive the rise in genomics-based analysis software and services segment.


Several companies, like IBM, Sun Microsystems and SGI are actively working towards developing IT as a tool to expedite research and discovery. The market for IT infrastructure and other services would also continue growing, in line with the overall market. The report, produced by Business Communications Company, costs US$3,950.


Contact: Business Communications Company Inc., #25 Van Zant Street, Norwalk, CT 06855, United States of America. Tel: +1 (203) 853 4266; Fax: +1 (203) 853 0348


E-mail: info@bccresearch.com


Website: www.drugresearcher.com

Pfizer enters VR1 deal with Renovis

In the United States, Pfizer and Renovis have signed a collaboration and licensing agreement to develop and commercialize small molecules that target a receptor, which generates signals for certain types of pain. This work also focuses on treatments for urinary incontinence and other diseases and disorders, targeting the vanilloid receptor, VR1.


Key mediators of pain signalling are ion channels, which regulate the flow of different ions (charged atoms) between the inside and outside of neurons. The transient receptor potential ion channels constitute a large and diverse family, several of which are thought to mediate pain signalling and are attractive targets for drug discovery. The best known of these is the vanilloid receptor 1 (VR1). A drug that blocks VR1, preventing it from activating nerve cell signalling, could provide a non-narcotic analgesic and would also be useful for treating non-neurological conditions such as inflammatory bowel disorders and asthma.


Under the terms of the agreement, the two companies will combine R&D on VR1, including all existing VR1 antagonists. Pfizer will have exclusive worldwide rights to commercialize products that result from the collaboration. Pfizer will fund all aspects of research and pre-clinical efforts to the tune of US$7 million as well as pay US$10 million as licensing fee to Renovis. It has the option to extend the agreement by up to two years, subject to additional funding needs.


Renovis will get more than US$170 million in royalties and milestone payments for each product resulting from the collaboration.


Website: www.drugresearcher.com

GENOMICS

Scientists find new genes in X-chromosome

A team of scientists from Institute of Bioinformatics (IOB), Bangalore, India, and Johns Hopkins University, the United States, has deciphered the human X-chromosome leading to identification of dozens of genes. According to Dr. Akilesh Pandey, IOBs Chief Scientific Adviser, this is the first time that any institute has provided an independent analysis of any human chromosome and the first real large-scale bioinformatics effort on human genome from any research organization in India.


For 18 months, 26 scientists of the team poured through the publicly available sequence of the X-chromosome to identify genes and other important parts of its DNA. By comparing human X-chromosome to genetic information from chimpanzees, rats and mice, the team uncovered dozens of new genes, many of which were located in regions of chromosome already tied to various diseases. The results also confirmed the existence of some of the newly identified genes.


Chemical Weekly, 10 May 2005

Link between ICAM1 gene and breast cancer reported

Scientists from Sequenom, California, the United States, have reported the involvement of intercellular adhesion molecule gene, ICAM1, in breast cancer metastasis. This corroborates the earlier finding that genetic variations in the ICAM gene region are associated with the occurrence of metastasis and establishes a causal role of ICAM1 in invasion of metastatic human breast cancer cell lines.


The researchers demonstrated that the level of ICAM1 protein expression on the cell surface positively correlates with metastatic potential of several human breast cancer cell lines and that ICAM1 mRNA levels are elevated in breast tumour compared with normal tissue. They also observed that ICAM1 down-regulation at the mRNA and protein level led to a strong suppression of human breast cell invasion. According to Dr. Charles Cantor, Chief Scientific Officer of Sequenom, the discovery of ICAM1 might help direct not only diagnosis, but importantly, therapeutic decisions.


Website: www.bio.com


Indians genes make them vulnerable to AIDS

Scientists led by Dr. N.K. Mehra, Department of Transplant Immunology and Immunogenetics, All India Institute of Medical Sciences, New Delhi, India, have found that genes associated with faster progression of AIDS are more common in people of India. The protective genes, which are associated with delayed onset of AIDS symptoms in HIV-infected people, are rare or absent in Indians. The team studied about 1,000 healthy people without HIV infection and over 200 HIV-infected people and found that PX, a gene that is known to be associated with rapid progress to AIDS, was 2.5 times more common in Indians than a protective genes called PY. PX and PY are two forms of a gene called HLA-B*35, which belongs to a family called Human Leukocyte Antigen (HLA) that modulates immune response against infection. A large number of genes are present in HLA family in Indians in comparison with Caucasians, which is attributed to racial mixture or to microbial pressure as Indians are faced with several infections.


The team also studied Chemokines, the group of genes that control entry of virus into macrophages that destroy the infection. The Chemokine CCR5 exists in five forms, of which 0201 associated with rapid progression to AIDS is very common with almost 35-40 per cent of Indians carrying it while the protective 0202 is rare. Similarly, a mutant form of CCR2, which is associated with rapid progress to AIDS, is carried by 35-40 per cent of people while only 15 per cent of people had the protective mutation.


According to Dr. Mehra, work is going on to find whether groups of genes that were found in 60 per cent of diabetics in India, and thus considered to increase susceptibility to the disease, give any protection against AIDS. The possibility exists because a different group of genes, which raises susceptibility to diabetes among westerners, has been found to bestow protection against AIDS.


Chemical Weekly, 10 May 2005

Evidence of selection in apoptosis genes

Scientists led by Dr. Rasmus Nielsen of Cornell University, the United States, have reported the strongest evidence of positive selection since divergence in genes that relate to immunological defence and apoptosis in human and chimpanzees. They found over 13,000 annotated human genes and their chimp equivalents, using the ratio of synonymous to non-synonymous mutations as evidence of positive selection, to determine which genes have been changing as humans and chimpanzees evolved into their modern forms.


Dr. Nielsens team subjected orthologous humanchimp gene pairs from published human data and polymerase chain reaction data obtained from a single male chimpanzee to likelihood analysis, which revealed any differences in mutation type ratio. Genes with fewer than three nucleotide differences were excluded. Dr. Nielsens team made several surprising discoveries, including increased positive selection on the X-chromosome and a relative lack of positive selection on genes expressed in the brain. Many genes that showed signs of positive selection were involved in sensory perception or immune defences, and the genes involved in apoptosis showed the strongest evidence for selection. During spermatogenesis, apoptosis kills a large proportion of cells. A mutation allowing a cell to avoid such a fate should spread in the population because it would confer a selective advantage. However, as cancer cells were also eliminated by apoptosis, any positive selection would be balanced against an increased risk of cancer.


Dr. Justin Fay, a geneticist at Washington Universitys School of Medicine, says that the statistical modelling technique of Dr. Nielsens team has limitations since it is only able to detect positive selection on a gene when it has multiple selected substitutions. It is entirely possible that every gene has had a single positively selected change between human and chimp. The sequences of many genes responsible for crucial differences between the two species can differ by only a single base, but would be missed by Nielsens method. Dr. Nielsen has acknowledged the limitations, saying that humans and chimpanzees were so alike that sometimes enough differences cannot be detected to make meaningful scientific statements, but many scientists were surprised to see how much one could determine about the evolution of these two closely related species.


Website: www.genomebiology.com

HIV-1 induces RNA silencing

A team of researchers led by Dr. Kaun-Teh Jeang and Dr. Yamina Bennasser of National Institute of Allergy and Infectious Diseases, Maryland, the United States, has found that natural small interfering RNA (siRNA) elicited RNA silencing in human cells. A sequence was characterized in the HIV-1 genome that encodes a rare siRNA precursor, a short hairpin RNA (shRNA) that was processed by Dicer, a cytoplasmic enzyme, into small siRNAs. The also virus prevented RNA silencing through a suppressor present in its Tat protein, which interferes with Dicers activity. According to Dr. Jeang, the holy grail of HIV researchers is the ability to find regions in the HIV genome that cannot mutate, and the sequence that he and colleagues characterized seems to be one such region that cannot change for functional reasons. There are, however, some dissenting opinions.


According to Dr. John Rossi of City of Hope Beckman Research Institute, the United States, the Dicer would be inhibited by Tat, which is exclusively a nuclear enzyme, leading to non-specific effects but the 19 base-pair hairpins are not substrates for Dicer. Dr. Mario Stevenson, University of Massachusetts, is of the opinion that RNA silencing can also be achieved in the nucleus, not only in the cytoplasm.


Website: www.genomebiology.com

MEDICAL BIOTECH

Korean team lauded for stem-cell advance

A team of scientists led by Dr. Woo Suk Hwang of Seoul National University, Republic of Korea, has made cell lines that are tailored to individual patients. Scientists hope that by studying these lines, they will be able to watch how diseases unfold, and learn how they might be cured.


The team has produced 11 human embryonic stem cell lines from 11 cell donors including men, women and children some with diseases such as diabetes or injuries to the spinal cord. A cell line has exactly the same genetic sequence as its donors own cells. As stem cells could be prompted to grow into any type of human cell, they could reveal clues about how each patients illness developed. For instance, by allowing the stem cells to develop into the cells that produce insulin, scientists might discover the signals that trigger their destruction in diabetics. Such patient-specific stem cell lines have never been created through cloning before.


Dr. Hwangs current study has also improved on the cloning procedure he reported last year. In his previous research, his team harvested 242 eggs from 16 different women to house the nuclei from the donor skin cells. After collecting all those eggs, only one stem cell line could be made. This time, the team says it used an average of 17 eggs to make each line, a major improvement in efficiency. This is important because the procedures that stimulate women to produce egg, and the techniques for removing them, carry health risks and could cause discomfort.


Website: www.nature.com

Nautilus announces interferon gamma progress

France-based Nautilus Biotech has developed a set of molecules single mutants of IFN-gamma which, developed as a biopharmaceutical, would allow less frequent injections of the drug. IFN-gamma has been marketed for the treatment of chronic granulomatous disease and malignant osteopetrosis, and was in clinical trials for the treatment of idiopathic pulmonary fibrosis, certain forms of cancer and tuberculosis. Experimental studies have shown that IFN-gamma has higher resistance to blood and intestinal proteases, while maintaining the native level of biological activity.


According to Mr. Manuel Vega, Chief Executive Officer of Nautilus, achieving higher stability is a key objective for the improvement of IFN-gamma, as it would increase the half-life of the molecule in the body and decrease the frequency of repeat treatments. It is also one of the most important clinical criteria for any next generation IFN-gamma with the potential to become the next generation of blockbusters.


Website: www.drugresearcher.com

Nanoshells that detect and destroy cancerous cells

Scientists led by Dr. Rebekah Drezek and Dr. Jennifer West, Rice University, Texas, the United States, have developed a new approach to fight cancer based on nano-scale particles that could both detect and destroy cancerous cells. Current imaging approaches only detect the cancer but do not offer a method of treatment.


The nano-particles used are nanoshells tiny spheres of silica coated with a thin layer of gold that exhibit unique size-dependent behaviour such as tunable optical properties, which allow designing of particles that scatter and absorb light at particular wavelength. The scattering of light provides the optical signal employed to detect the cancer cells, which then light up when they come into contact with the nanoshells. In this study, the researchers designed the nanoshells to look for breast cancer biomarkers on the surface of the cancer cells. The technique can be extended to target other types of cancer or disease processes that have known surface markers. The additional ability of the particles to absorb light is used to generate heat, which then destroys the cancer cells.


Express Pharma Pulse, 21 April 2005

Liver cells make insulin

Dr. Sarah Ferber and colleagues at Sheba Medical Centre in Tel-Hashomer, Israel, have shown that adult human liver cells can be made to produce insulin and transplanted them into mice to treat diabetes. This discovery would allow diabetics to donate their own insulin-producing tissue and such treatments could overcome problems with current cell replacement therapies for diabetes, which were hindered by limited availability of donor tissue and the need for immune suppression.


The researchers were investigating a treatment that involves tweaking specific liver cells that express homeobox gene-1, which is known to play a role in the development of the pancreas and in the function of insulin-producing cells. Homeobox gene-1 activated the insulin promoter in up to 25 per cent of treated liver cells, and insulin was stored and secreted in a way similar to that in pancreatic -cells. To test the cells, they were transplanted into hyperglycemic mice with diminished insulin levels and it was found that the cells caused a gradual and prolonged decline in blood glucose levels.


Website: www.betterhumans.com

Novel way to deliver continuous attacks on cancer cells

A team of researchers led by Prof. David Mirelman at Weizmann Institute of Science, Israel, has found a way to deliver a powerful punch against cancer by conjugating rituximab, an antibody already in clinical use, with allicin, the product of an interaction between alliinase enzyme and the chemical alliin. Alliin occurs naturally in plants such garlic and onion as a defence mechanism against soil fungi, bacteria and parasites.


Allicin molecules could easily penetrate biological membranes and kill cells. Rituximab is designed to target and lock on to the surface of certain types of cancer cells such as lymphoma. When administered alone, rituximab serves as a marker and docking point for the bodys own immune system to kill the cancer cell. Cancer cells can be destroyed more efficiently by arming this antibody. The researchers first chemically bound alliinase to rituximab and then injected this conjugate into mice that had been implanted with human lymphoma cancer cells. This armed antibody found and bound to target cancer cells. Subsequently, the mice were repeatedly injected with the inert chemical alliin that, upon contact with alliinase, was processed into allicin molecules directly on the cancer cells surface. Within three days, almost all human lymphoma cancer cells were destroyed in those mice treated with the conjugate and alliin, while hardly any cancer cell destruction occurred in the control mice that received the conjugate alone. The production of allicin could be turned off by the ceasing the administration of alliin.


Chemical Weekly, 24 May 2005

Virus vaccines show promise

A team of researchers led by Dr. Steven Jones of the National Microbiology Laboratory, Canada, and Dr. Thomas Geisbert, Virologist at the United States Army Medical Research Institute of Infectious Disease, has developed vaccines that protect monkeys from two deadly viruses Marburg and Ebola. This experimental vaccine could be safe and effective also for humans exposed to the contagious viruses, which are almost always fatal.


There is no vaccine or treatment other than drugs to relieve pain for victims. Marburg and Ebola viruses spread by bodily fluids such as blood, sweat and saliva. Most victims die within days after massive bleeding. Researchers replaced a protein in an animal virus with a protein from the Ebola and Marburg viruses, and successfully tested the vaccines on rodents. Following this, monkeys were injected first with the vaccines and 28 days later with the viruses. The virus-injected monkeys survived. As monkeys suffer almost the identical disease to humans, the researchers are confident that this approach would work in humans.


The new Ebola vaccine has advantages that it is a live vaccine that grows inside the recipient for a short period of time, generating a rapid and strong immune response, and does not require revaccination every year. Researchers will now produce the vaccines in a regulated facility, and repeat the studies to ensure they work, are safe and determine how quickly the vaccines take effect.


Website: www.wired.com

Scientists infuse rats with human stem cells

Researchers led by Dr. Clive Savendsen, Professor of Anatomy at the University of Wisconsin-Madison, the United States, have been able to direct certain type of new stem cells to secrete a neuro-protecting protein by inserting engineered human stem cells into the spinal cords of rats afflicted with amyotrophic lateral sclerosis (ALS) before injecting them back into the rat spinal cord.


The scientists worked with specialized neural stem cells known as neural progenitor cells that arise from primitive stem cells during the first few weeks of human brain development. According to Dr. Savendsen, neural stem cells are much more appropriate for clinical use because, unlike embryonic stem cells, they can grow in the absence of animal derivatives that are considered a potential source of contamination. Once inside the brain or spinal cord, neural progenitor cells grow into neuron-supporting stem cells called astrocytes. Researchers believe that ALS causes malfunction in astrocytes, eventually causing their death.


Express Pharma Pulse, 28 April 2005

Scientists create plant factories for antibodies against tumour

Scientists create plant factories for antibodies against tumour Dr. Hilary Koprowski, Dr. Kisung Ko and colleagues at the Jefferson Medical College of Thomas Jefferson University, Philadelphia, have produced a monoclonal antibody against colorectal cancer by inserting into tobacco plants DNA coding for an antibody. The plants, in turn, became factories churning out the antibody at cheaper and faster rate, raising hopes that this technology could be used in humans.


Standard mouse-made monoclonals recognize a particular type of protein antigen on human colorectal cancer cells and have been used in treating metastatic disease and in preventing recurrence in certain high-risk patients. But large-scale production was expensive. The scientists had previously shown that tobacco plant-made monoclonal antibodies could neutralize rabies virus and prevent disease in infected mice and also that plant-made monoclonal antibody purified from tobacco leaves could recognize, or bind to, human colorectal cancer cells. Next, they grafted human colorectal cancer cells onto the backs of nude mice mice stripped of their immune systems and subsequently injected the animals with the plant-derived antibodies. The scientists found that tumour growth was inhibited in a similar manner to that of mammal-made monoclonal antibodies. These results show that plant biotechnology could be a useful, safer and less expensive way to produce monoclonal antibodies.


Website: www.jeffersonhospital.org

Enzyme block sparks heart muscle repair

A team of researchers led by Dr. Mark Keating and Dr. Felix Engel at Howard Hughes Medical Institute, Boston Childrens Hospital and Harvard Medical School, the United States, has found that blocking a specific enzyme sparks the growth of heart muscle cells and could provide an alternative to stem cell therapy for treating heart attack damage.


The researchers have shown that inhibiting an enzyme called p38 MAP kinase enables heart cells to proliferate. The enzyme normally suppresses cardiomyocyte replication. Cardiomyocytes have been thought to be incapable of replicating in mammals after birth. This is of particular concern after a heart attack, when dead heart tissue does not regenerate. But foetal rats cells showed that greater p38 activity correlates with reduced cardiac growth while reduced activity correlates with accelerated growth. The researchers also showed p38s role in cell replication using adult cardiomyocytes. Inhibiting p38 in cultures of cardiomyocytes from rats was shown to increase DNA synthesis. P38 was also seen to regulate the activity of genes required for mitosis. Furthermore, mice bred to lack p38 had increased mitosis by more than 90 per cent in their cardiomyocytes. Finally, inhibiting p38 promoted cytokinesis.


Website: www.betterhumans.com

PROTEOMICS

Protein that could stop replication of cancer cells

Researchers led by Dr. Chung In-Kwong of Yonsei University, Seoul, Republic of Korea, and Dr. Mark Muller of University of Central Florida, the United States, have found that the protein called MKRN1 promoted the destruction of an enzyme called telomerase that enables rapid duplication of cells.
A long stretch of repeated DNA, called telomere, has a role in influencing cell length and, in turn, its lifespan. Each of the humans 46 chromosomes is capped on either end by telomeres, which help protect the cells.


Each time a cell divides, the telomeres are shortened until eventually they turn so small that the cell stops multiplying. Eventually the cell is eliminated from the body. When telomere ends do not shorten, division continues unabated. In laboratory tests, the MKRN1 protein eliminated the presence of telomerase in tumor cells. The researchers found that many different species have these genes, emphasising their important collective roles in life. Moreover, mutating or altering the MKRN1 gene is lethal; cells cannot live without these genes. The effectiveness of MKRN1 is greatly increased when combined with the drug geldanamycin, disrupting the formation of cancerous tumours by binding with protective proteins.


Website: www.bio.com

Protein essential for hearing also vital for pain perception

A team of researchers led by Dr. Jaime Garcia-Aoveros, Northwestern University Feinberg School of Medicine, Michigan, the United States, has found that the protein known as TRPA1, which translates sound into nerve signals to the brain and enables individuals to hear, is also required for pain perception. TRPA1 is found in the majority about 75 per cent of the bodys pain-perceiving neurons but not in major organs. Drugs that block TRPA1 would be novel painkillers with hardly any side effect.


Besides being expressed in nociceptors (pain neurons), TRPA1 is present in the stereocilia of hair cells, used for hearing as well as detecting gravity and maintaining balance, and in the organ of corti, the hearing organ of the inner ear. The researchers have found properties of the TRPA1 channel that accounted for its suspected parallel role in pain sensation, such as why in some cases pain from an injury will not go away as long as the injury remains. TRPA1 opens in response to painful stimulation and ions enter the cell, making it less negatively charged, or depolarized. At this point, most ion channels close, a phenomenon known as inactivation, because their signalling task has been achieved. But TRPA1 senses the depolarization and responds to it by staying open, closing only when the harmful stimulus stops or the depolarization is small.


TRPA1 may account for painful phenomena in damaged tissues such as lack of desensitization or even some forms of enhanced sensitization, such as hyperalgesia (extreme sensitivity to pain) or allodynia (pain resulting from non-noxious stimuli to normal skin). According to Dr. Garcia, TRPA1 channel blockers could be looked for novel analgesics that block pain at its initiation which should probably be used topically preferably avoiding contact with the inner ear but could also be applied systemically. In that case, the patient might have to put up with temporary deafness, a side effect that is probably better than extreme pain.


Website: www.eurekalert.org

Researchers uncover structure of key protein complex in cells

Dr. Christopher Lima and co-researchers at the Memorial Sloan-Kettering Cancer Centre, New York, the United States, have uncovered the structure of a network of proteins that help regulate the life cycle of cells. This would help in learning its precise function, and in finding ways to correct flaws in the system that could lead to cancer.


Using X-ray crystallography, the researchers identified how one protein, a molecule called the Small Ubiquitin-related Modifier (SUMO-1), is connected or conjugated to another molecule within the complex. Understanding this process could be important in many physiological steps that are disrupted in cancer including error-prone DNA replication, chromosome segregation, the cells response to stress and normal signalling reactions as SUMO-1 is necessary for the normal assortment of chromosomes immediately before a cell splits. For instance, yeast cells that lack SUMO cannot successfully divide. According to Dr. Lima, since unchecked cell division characterized cancer, proving SUMOs function could potentially yield novel anti-cancer therapies. If sumoylation (SUMO attaching to a protein) was promoting cell division, then SUMO inhibitors may be agents that block the cell cycle.


SUMO-1 did not appear to label other proteins for destruction. It instead appeared to alter their function in different ways, perhaps even by preventing ubiquitin tagging. Still, there were enough similarities between conjugation of SUMO and ubiquitin that the latest findings could help researchers understand both. The structural network revealed a mechanism that may be important in the regulation process in the SUMO and ubiquitin pathways, pathways that need to operate properly for normal cell division. The complex described includes SUMO-1, a substrate, and two other proteins called E2 and E3. E2 links SUMO to target molecules, while E3 is a kind of chemical marriage broker, bringing targets close enough to SUMO to facilitate binding.


Website: www.bio.com

Enzyme group that converts protein into cellular traffic signal

Researchers led by Dr. Jacek Gaertig of University of Georgia, the United States, and Dr. Bernard Edd of National Centre for Scientific Research, France, have identified a new group of enzymes, called polyglutamylases, that may help uncover how cells direct internal traffic. Polyglutamylases attach glutamic acid (an amino acid) chains of varying length and branching patterns. The discovery has future implications for conditions such as polycystic kidney disease, behavioural disorders, male infertility and cancer that involve defects in protein fibres called microtubules.


The newly identified enzyme group attaches an unusual molecular tag to a component of the cells microtubular highway system. The tag is attached to a localized region of a microtubule and may act like a road sign, directing motor proteins to take the proper exit to the nucleus or the cell membrane. Strings of glutamic acid are sometimes attached to the side of a protein called tubulin, which is a component of microtubules.
The research team identified the active subunit of the enzyme complex and found genes for these enzymes in many organisms including humans. These enzymes could modify just a portion of a microtubular highway, an important discovery that suggests the mechanism for directing cell traffic. This research paves the way of detailed studies of what polygultamylate modification does and how it works.


Website: www.eurekalert.org

Scientists observe infectious prions in action

Scientists led by Dr. Byron Caughey, Rocky Mountain Laboratories (RML) in Hamilton, the United States, and Dr. Marco Prado, University of Minas Gerais in Belo Horizonte, Brazil, have for the first time watched agents of brain-wasting diseases, called transmissible spongiform encephalopathies (TSE). TSE diseases, caused by certain protinaceous infectious particles called prions, include scrapie in sheep and goats, mad cow disease in cattle, chronic wasting disease in deer and elk, and Creutzfeldt-Jacob disease in humans. The scientists performed the research in laboratory cultures using a rodent-adapted form of scrapie protein and cells taken from the central nervous system of mouse and hamster brains. The proteins were first branded with fluorescent dyes.


Researchers have tracked prions moving through other parts of animal bodies up to the brain, but none had ever tracked the protein movement within animal brain cells. One of the most difficult aspects of the experiment was finding a way to put a fluorescent tag on the TSE prion proteins without altering them, while still allowing identification of the prions as they penetrated and spread within the nerve cells. This is technically difficult because the scrapie pathogen is largely a corrupted form of a host cell protein, and it is hard to detect and distinguish it from its normal counterpart. Once the researchers learned how to mark the prion proteins, they added them to a culture of nerve cells and then began watching and taking photo images with a confocal microscope. The photos that, when put into a video format, showed prion protein moving within cells, then along narrow cellular projections called neurites and ultimately into close proximity with adjacent cells. These findings offer leads towards developing therapies to stop the spread of TSE.


Website: www.bio.com

AGRI BIOTECH

Researchers knock out a gene in potato to procure starch

Research led by Dr. Jost Muth, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Germany, has developed a technique that helps potatoes produce special starch usable in the pulp and paper industry. Potatoes contain two types of starch, amylose and amylopectin, each with a different molecular structure and hence different properties.


Genetically modified tubers with only amylopectin do exist, but the variety is yet to be approved for agricultural use. IME researchers treated seed potatoes with ethyl methane sulfonate, a substance commonly used by plants breeders, to knock out a single gene and thus prevent potatoes from producing amylose. This treatment caused random point mutations in the plant DNA. The scientists then analysed the mutations and selected plants containing a damaged amylose gene for further cultivation. Only potatoes with two copies of the defective amylose gene go on for further reproduction, in order to retain the mutation. The scientists hope that amylose-free potatoes could be made available within few years as commercial crops.


Chemical Weekly, 10 May 2005

Drought-resistant GM rubber

Scientists at the Rubber Research Institute of India (RRII) in Kottayam, Kerala, have developed genetically modified rubber plants that have increased drought resistance as well as environment stress tolerance. They selected the popular RRII 105 variety for the experiments and introduced four specific genes that would provide drought tolerance, tapping panel dryness tolerance and elevated temperature and light tolerance. These genes were introduced into rubber tissues separately, and transgenic plantlets were developed with the gene coding for superoxide dismutase (SOD), hardened and transferred to polythene bags. These plantlets were multiplied through bud grafting.


Preliminary biochemical studies revealed that the SOD transformed tissues over-expressed the gene when subjected to artificial stress conditions. According to Dr N.M. Mathew, Director of RRII, the major objective of genetic transformation of rubber trees at the RRII was the introduction of genes controlling specific agronomic traits such as the genes for resisting diseases, drought and other environmental stress tolerance, enhanced rubber biosynthesis and timber yield, and tolerance to tapping panel dryness to high yielding rubber clones. The transfer of selected genes in a single generation by genetic transformation is especially interesting, as the rubber trees improvement through conventional breeding is limited by long breeding cycles and high heterozygosity.


The institute has now initiated efforts to develop transgenic rubber plants with enhanced rubber production by over-expressing the genes involved in the rubber biosynthetic pathway. Research is also in progress to develop transgenic plants producing pharmaceutically and industrially useful recombinant proteins in the latex.


Website: www.hinduonnet.com

Plants use dual defence system to fight pathogens

Dr. David Mackey and co-researchers at Ohio State University, Columbus, the United States, have uncovered the link between two biochemical pathways that plants use to defend themselves against pathogens pathways that scientists have long believed worked independently of each other.


The researchers infected Arabidopsis plants with a bacterial strain of Pseudomonas syringae, a bacterium that usually infects tomato crops. One of the immune pathways that they recognized is pathogen-associated molecular patterns or PAMPs. The PAMP pathway appears to be a plants first line of defence against pathogens. The other pathway uses disease-resistant proteins, or R-proteins, which could detect certain molecules, called effectors, that are secreted by pathogens. This pathway produces a stronger immune response than the PAMP pathway. Certain types of bacteria, including P. syringae, make a hypodermic needle-like structure that pierces the outermost membrane of a healthy plant or animal cell. The pathogen uses this conduit to send infectious effector proteins into the host cell. While P. syringae injects about 40 different varieties of effector molecules into a plant cell, the researchers focused on the actions of two of these molecules AvrRpt2 and AvrRpm1. Both target a protein key to Arabidopsis health.


The researchers found that both AvrRpt2 and AvrRpm1 effector molecules effectively shut down the PAMP pathway. But the plants R-proteins detect this and come to the rescue. PAMP defence responses are often effective but may be blocked by the pathogens effector proteins. If an R-protein identifies a pathogens presence, it usually induces a very strong immune response, in most cases stopping a would-be infection. A next step in this line of work is to look at other pathogen effector proteins and analyse their role in causing infections.


Website: www.researchnews.osu.edu 

Researchers identifies gene suspect for potato blight

A research team led by Dr. Sophien Kamoun at Ohio State Universitys Ohio Agricultural Research and Development Centre in Wooster, the United States, has found a gene called Avr3a, the first avirulence (Avr) gene identified from Phytophthora infestans, the plant pathogen that causes late blight. P. infestans contains a diverse set of such avr genes that, depending on the plant variety, can either facilitate disease or trigger resistance. It belongs to a group of destructive pathogens called oomycetes, which look like fungi and algae.


Over the last 75 years, potato breeders have introduced at least a dozen late blight-resistant genes into the cultivated potato. The researchers thought that if a potato plant contained one of these genes, R3a, then it could detect P. infestans manifestation by recognizing Avr3a and ward off the impending disease. They found that the leaves containing the R3a gene successfully resisted late blight infection when exposed to P. infestans races that carried the Avr3a gene. However, some races of P. infestans with mutations in their Avr3a gene escaped the resistance response triggered by R3a. Laboratory analysis showed that when R3a detected Avr3a in a leaf cell, that plant cell died.


According to Dr. Kamoun this programmed cell death is how the plant keep the pathogen from spreading to other cells, since only a few cells die, and is one defence mechanism that some plants have. Until now, researchers knew very little about P. infestans at the molecular level. By studying Avr3a, R3a and similar genes, researchers may be able to determine what happens during the first stages of late blight infection.


Website: www.researchnews.osu.edu  

Plant pest resistance boosted

A team of researchers led by Dr. Paul Christou at the University of Lleida, Spain, has developed a new technique for increasing pest resistance in transgenic crop plants. The strategy, which boosts and broadens the activity of Bacillus thuringiensis (Bt) toxins, targets previously impervious pest species and reduces by up to thousand-fold the level of toxin expression needed.
Researchers devised a new strategy that increases the repertoire of toxin-binding sites that a Bt toxin attacks.


They fused the sequence for Bt toxin Cry1Ac with that of the nontoxic B-chain subunit of ricin (RB) in a recombinant plasmid. RB is a leptin that binds with galactose as well as N-acetylgalactosamine residues with high affinity, the latter of which are key components of Bt toxin-binding receptors. Then embryonic callus from mature maize seeds were bombarded with this BtRB fusion. The researchers tested their fusion toxin against stem borer Chilo suppressalis, a pest normally susceptible to Cry1Ac, and found that maize producing low levels of BtRB killed 75 per cent of larvae, compared with 17 per cent in Bt-only plants. Similar trials with the cotton leaf worm Spodoptera littoralis, which is resistant to Bt delta endotoxins, showed that after 4 days, nearly 78 per cent of larvae died on BtRB maize, compared with less than 20 per cent on Bt-only or nontransformed maize. In the leafhopper Cicadulina mbila, which like other homopterans was ordinarily not affected by Bt toxins, 95 per cent of insects died by day 4 on BtRB maize, compared with 80 per cent survival otherwise. Tests with the homopteran cereal aphid Rhopalosiphum padi, however, did not show any toxicity with BtRB or Bt-only maize.


Website: www.thescientist.com

PUBLICATIONS

Bioinformatics and Functional Genomics

Bioinformatics and Functional Genomics provides an important integrated approach to understanding both the theory and practice of the emerging field of bioinformatics, covering areas from sequence analysis, gene expression, microarrays and proteomics to molecular phylogenics and whole genome studies. Part 1 covers analysing DNA, RNA and protein sequences in databases, Part 2 discusses areas related to genome-wide analysis of RNA and protein and Part 3 focuses on genome analysis.

The book emphasizes practical skills, such as how to analyse genes and proteins, how to make trees using phylogenic software, how to extract data, and how to identify genes and proteins implicated in disease. Although the text emphasises the use of computational tools and databases, it assumes no computational background, which make it an ideal reference at every level of study.

Tissue Engineering: Essentials for Daily Laboratory Work

Comprehensive in its scope and illustrated in detail, this practical book provides a basic insight into the complex world of tissue development and artificial cell culture using tissue engineering. The introductory chapters cover basic cell biology and cellular development as well as cell culture, with an emphasis on ways of differentiating tissue and the critical evaluation of the properties of maturing tissue constructs. The book also focuses on the use of stem cells from the most varied sources in tissue engineering. It also has an exceptionally wide-ranging glossary containing some 1,000 key words from the fields of cell biology, cell culture development and tissue engineering.


For the above publications, contact: John Wiley & Sons Ltd., Journals Fulfillment, 1 Oldlands Way, Bognor Regis, West Sussex PO22 9SA, United Kingdom. Tel: +44 (1243) 843335; Fax: +44 (1243) 843232


E-mail: cs-wis@wiley.co.uk


ASIAN AND PACIFIC CENTRE FOR TRANSFER OF TECHNOLOGY

This website is optimized for IE 8.0 with screen resolution 1024 x 768
For queries regarding this website, contact us
Copyright © 2010 APCTT | Privacy Policy | Disclaimer | Feedback