VATIS Update Biotechnology . Jul-Sep 2014

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Biotechnology Jul-Sep 2014

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
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Biotech strategy in India

The National Biotechnology Development Strategy — 2014 (Biotech Strategy II) proposes to strengthen industrial biotechnology in India with a vision to produce 20% of India’s bulk chemicals by biocatalysis/bioprocessing. The Biotech Strategy II was issued recently by the Department of Biotechnology (DBT) to establish India as a world class bio-manufacturing hub for developing and developed markets. To achieve this vision, the DBT during the next years will set up five technology platform centres. This could be achieved by setting up multi-disciplinary centres around existing academic centres of excellence in collaboration with industry. Besides, the DBT will set up technology development centres within existing academic institutes. These centres will work closely with industries and pilot test the technologies developed by the centres or others at a reasonable scale and engage desirous industry to assist them to take the developed technologies to commercialisation.

Revival of timber industry in Malaysia

The Brunei, Indonesia, Malaysia and the Philippines East Asean Growth Area (BIMP-EAGA) Malaysia Business Council (BEMBC) is working closely with a biotechnology company to revive Sabah’s timber industry. The Council has been working on sustainable forest timber farming of light hardwood timber with BioJadi Technology Sdn Bhd. Certain species of the light hardwood tree, which requires a gestation period of between 20 and 25 years to mature can be harvested in seven years using a new biotechnology developed by BioJadi Technology. For this, any type of land is suitable except peat soil, where one hectare can be systematically planted with 625 trees. The cost of planting is about RM10,000 for the entire duration and includes land clearance, planting materials and seedlings, fertilisers and care for the trees, until maturity and ready for harvest at the end of the seventh year. The gross profit from harvesting is about 12 times the cost of investment. BioJadi, a locally developed biotech and organic fertiliser, when applied throughout the trees’ growing years can make them ready for harvesting in just five years and is suitable for reforestation.

Zoonotic diseases ignored in developing world

Anthrax, brucellosis and bovine tuberculosis impact greatly on human and animal health in developing nations. These diseases — known as zoonotic diseases — are spread between animals and humans, and are common in societies where poverty is widespread, and where people depend on animals for their livelihood. These diseases have been eliminated or brought under control in more developed countries, as simple and effective controls are available, whereas they have been neglected in developing countries. Poor healthcare infrastructure in affected countries can often mean that thousands of sufferers are left un-diagnosed. This presents huge challenges to health professionals, policy makers and researchers in their efforts to combat the diseases. Chikungunya, dengue, avian influenza, plague, SARS and acute encephalitis syndrome (AES) are some of the zoonotic diseases that have taken and continue to take a heavy toll on human life in India. Mortality and morbidity due to Japanese encephalitis and AES in the eastern parts of India is high. Reports of deaths due to chikungunya, dengue and highly infectious Congo haemorrhagic fever are also not uncommon, particularly during monsoon.

Portable device test for TB is faster and cheaper

A new low-cost 30-minute test for tuberculosis that is not only faster but more accurate than tests currently in use has been developed. It could help control the disease by detecting it early. Jeffrey Cirillo, professor at the Texas A&M Health Science Center College of Medicine, and investigators at Stanford University, the United States, have identified a new chemical compound to spot the bacteria that cause TB with a level of sensitivity that currently takes months to produce. Findings show the test can determine that a patient has tuberculosis with 86% sensitivity and 73% specificity. Using a fluorescent substrate, the device targets BlaC — an enzyme produced by the bacteria that cause TB — as an indicator of the bacteria’s presence. Once sputum samples are combined with the reactive substance, a battery-powered, portable tabletop device, the TB REaD, detects any fluorescence and delivers the diagnosis in as little as 10 minutes. Additionally, the one-step test will require little technical expertise or resources, should take less than 30 minutes to carry out, and is easily transportable, making it an ideal candidate for field diagnosis in developing countries.

Brazil approves GM mosquito that could cut dengue

The National Technical Commission on Biosecurity, the organisation that regulates transgenic organisms in Brazil, has decided that a genetically modified (GM) strain of mosquitoes whose offspring die before reaching adulthood do not pose a significant risk to humans or the environment. The GM strain, known as OX513A, has already been released in experiments across Brazil. Its developer, UK biotech firm Oxitec, plans to apply for a licence to sell it to local government bodies in Brazil. The male GM mosquitoes have two additional genes: one makes a protein that causes a breakdown in the insect’s development and a second that acts as a marker, enabling researchers to monitor the mosquitoes in the field. Wild female mosquitoes that mate with GM males transfer the genes to their offspring, which die before reaching adulthood. In three field trials in Brazil’s Bahia state since 2011, successive releases of the transgenic strain reduced the wild adult population of A. aegypti by between 79 and 93%. Implementing Oxitec’s technique in a town of 50,000 people would cost two to five million reals (about US$890,000 to US$2.2 million) in its first year and around one million reals in following years.

Kenya’s GM ban and the future of GM policy in Africa

Growing genetically modified (GM) crops is legal only in three African countries — South Africa, Sudan, and Burkina Faso. Following a study results published in a journal in September 2012 stating that GM maize consumption was associated with tumor growth in rats, President Kibaki signed into law a blanket ban that would prohibit the import, sale, distribution, or consumption of GM foods in Kenya. This sent out a shock wave of anxiety throughout sub-Saharan Africa, prompting President Goodluck Jonathan of Nigeria to postpone his country’s plans to allow field testing of GM crops, which would have otherwise preceded legalizing commercialization. This ban was severely detrimental to the process of rational policy-making in general. Without consulting any of its agricultural research institutions, the Kibaki administration bypassed the NBA and single-handedly shut down GM imports. In a blanket ban like Kenya’s, both Golden Rice, a vitamin-fortified rice variety which has been genetically modified to enhance nutritional value, and WEMA, a drought-tolerant maize project too would be swept in under one big umbrella with cash crops, Monsanto’s commercial Round-Up-Ready maize, and whatever other GM organisms inside or outside of African that strike fear into the hearts of environmental NGOs Sub-Saharan Africa claims the highest prevalence of malnutrition in the world, and undernourishment contributes to about a third of deaths in children under the age of five If Kenya and other sub-Saharan African countries intend to eradicate malnutrition and food insecurity, they will have to seriously rethink their agricultural systems, which involves capitalizing on new forms of agricultural technology. And if they want to tap into the potential of GM crops, they will have to adhere to rational policy-making processes that leave no room for sloppy decisions based on whims or unfounded science.

Simulated human heart used to screen drugs

Dr Helen Maddock of Coventry University, the United Kingdom, has developed a pioneering new way—using samples of beating heart tissue—to test the effect of drugs on the heart without using human or animal trials. The breakthrough could lead to the lives of hundreds of future patients being saved and the quality of their treatments improved. Dr Maddock’s in vitro technique uses a specimen of heart tissue attached to a rig allowing the muscle to be lengthened and shortened while being stimulated by an electrical impulse, mimicking the biomechanical performance of cardiac muscle. Trial drugs can then be added to the tissue to determine whether or not they have an adverse effect on the force of contraction of the muscle (and therefore of the heart). This ‘simulated’ cardiovascular system — known as a work-loop assay — provides the most realistic model of heart muscle dynamics to date, and opens up unprecedented possibilities for identifying negative effects of drugs early and inexpensively.
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Approval for world’s first malaria vaccine

GlaxoSmithKline (GSK.L) is applying for regulatory approval for the world’s first vaccine against malaria, designed for children in Africa. The shot, called RTS,S, is intended exclusively for use outside the European Union but will be evaluated by the European Medicines Agency (EMA) in collaboration with the World Health Organisation (WHO). Although scientists at GSK have been working on this one for 30 years, hopes that RTS,S would be the final answer to wiping out malaria were dampened when results from a final-stage trial in babies aged six to 12 weeks showed that the shot provided only modest protection, reducing episodes of the disease by 30%. Data from that and other final-stage Phase III trials — conducted at 13 African research centers — have also been included to support the application. WHO has previously indicated it may recommend use of RTS,S from as early as 2015 if EMA drugs regulators back its license application. The vaccine contains an adjuvant, or booster, made by U.S. biotech company Agenus (AGEN.O). If approved, the vaccine is unlikely to be anything other than neutral for GSK’s bottom line. The firm has promised it will be priced at cost of manufacture plus a 5% margin.

Wheat draft genome sequence ready

An international consortium of scientists, including some from India, has put together a draft genome sequence for bread wheat (Triticum aestivum), providing an overview of its genetic make-up. Bread wheat arose from a series of hybridisation events that ultimately resulted in chromosomes from three ancestral species being merged. Consequently, the plant has ended up with three sets of seven pairs of chromosomes, each set forming a distinct sub-genome. Working out the plant’s genome required disentangling the sub-genomes with very similar chromosomes. The International Wheat Genome Sequencing Consortium accomplished the task by first physically separating the chromosomes. One chromosome from each of the 21 pairs was then sequenced to establish the exact order and location of units of DNA, known as bases, that it possessed. The bread wheat genome, 17 billion bases in size (five time larger than that of humans), has been analysed and some 124,000 genes identified. It also turned out that none of the sub-genomes is dominant with more of its genes being utilised to turn out proteins.

Indian scientists were given the responsibility of sequencing chromosome 2A. This chromosome alone has about 900 million bases. At present, only a ‘draft sequence’ is available for 20 pairs of chromosomes. A finished version, with minimal gaps, is ready for only one pair of chromosomes. The aim is to get the other chromosomes to that stage within three years. The chromosome-based sequence will help plant breeders learn how genes controlled complex traits such as yield, grain quality, disease, pest resistance and the ability to withstand various kinds of stress Having the complete wheat genome sequence in hand will allow scientists to activate genes that the plant possesses to create varieties producing better quality grain, which are more heat tolerant as well as capable of withstanding changing climatic conditions

Wearable sensor patch subsystem project

The Nano-Bio Manufacturing Consortium (NBMC) has selected a proposal by the University of Massachusetts, the United States, to develop a wearable sensor patch subsystem as its first project in its mission to develop human performance-monitoring electronics. The project has $450,000 in total funding.

This project focuses on the integration of the most mature system subcomponents in order to provide a deliverable demonstration of a microfluidic subsystem capable of time-gated acquisition of a sweat sample, along with detection of the target biomarker Orexin-A, which is a naturally occurring neuropeptide hormone released by the hypothalamus, and plays a crucial role in the stability of arousal and alertness. A wearable paper-based patch incorporating this technology would ideally provide valuable data regarding wearer fatigue.

This research could have profound implications not only for military use, but for any high-stress job in which alertness and responsiveness are critical: jet pilots, air traffic controllers, fire fighters, heavy-equipment operators, and in many other careers.

The project’s primary development centers on an interdigitated field effect transistor (FET) sensor. Detection of Orexin-A in blood and saliva samples has been demonstrated and this system offers the lowest-risk system integration.

India/US to begin collaborative research on diabetes

India and the US will soon begin bilateral collaborative research partnerships (CRPs) on diabetes. This collaboration is significant as in both the nations, diabetes is striking increasingly in younger age groups Specific areas of interest for this programme include studies that may address or be focused within one or more of the broad research areas such as prevention and management of diabetes or its complications; pathogenesis and pathophysiology of diabetes and its complications; diabetes in youth; innovative technologies for management and prevention of diabetes and/or its complications; and gestational diabetes. The CRP must be based on interactive relationships that maximise the expertise of the Indian and US research teams and interactions between their parent institutions and granting agencies. Indian and US investigators should work together to develop and submit applications to ICMR and NIH.

GAVI Alliance wants India to bolster immunisation programme

With a birth cohort of 26 million infants, India is home to the largest number of unimmunised children - 6.8 million or roughly a third of the world’s total. GAVI Alliance, a public-private partnership committed to saving children’s lives and protecting people’s health by increasing access to immunisation in poor countries, has called for renewed efforts in India to bolster routine immunisation coverage With GAVI Alliance support, India will continue the national roll out of the 5-in-1 pentavalent vaccine, which protects against diphtheria, tetanus, pertussis, hepatitis B and haemophilus influenza type B disease. GAVI will also provide US$ 107 million over three years, until 2016, towards health system strengthening in India targeting lower performing states. This will help India maintain the momentum and capitalise on the success of its polio campaign to strengthen routine immunisation nationally. The Alliance brings together developing country and donor governments, the World Health Organization, UNICEF, the World Bank, the vaccine industry, technical agencies, civil society, the Bill & Melinda Gates Foundation and other private sector partners. GAVI uses innovative finance mechanisms, including co-financing by recipient countries, to secure sustainable funding and adequate supply of quality vaccines.

Biotech industry forms international council

The Biotechnology Industry Organization (BIO) has reported that an international confederation of biotechnology trade associations has created the International Council on Biotechnology Associations (ICBA) to advocate public policies that support worldwide biotechnology growth. Formation of ICBA is an important step that allows us to better coordinate, organize and face issues around the world such as combating debilitating and rare diseases, reducing our environmental footprint, feeding the hungry, using less and cleaner energy, and having safer, cleaner and more efficient industrial manufacturing processes. The ICBA will advocate for more efficient and effective governance, science-based regulatory systems and the right framework conditions for competitiveness and innovation to ensure faster and more equitable access to biotech products and processes for patients, farmers and consumers around the world.

Stem cells can restore hearing ability

Scientists from Germany and the United States have found that spiral ganglion stem cells present in the inner ear can be used to restore hearing ability. Spiral ganglion cells are essential for hearing and their irreversible degeneration in the inner ear is common in most types of hearing loss. Adult spiral ganglion cells are not able to regenerate. Now, new evidence in a mouse model shows that spiral ganglion stem cells are capable of self-renewal and can be grown and induced to differentiate into mature spiral ganglion cells as well as neurons and glial cells. These properties demonstrated by spiral ganglion stem cells make them a promising source of replacement cells for therapies designed to regenerate the neural structures of the inner ear.


Genome Center for Infectious Diseases

Researchers from the J. Craig Venter Institute (JCVI), the United States, have won a five-year, up-to-$25 million grant from NIH’s National Institute of Allergy and Infectious Diseases (NIAID) to establish and operate a Genome Center for Infectious Diseases (GCID). The JCVI-led team will apply next-generation DNA sequencing technologies, as well as new bioinformatics and computing capabilities, with the intent of enhancing understanding of pathogen biology, virulence, drug resistance, immune evasion, and host microbiome biological interactions. GCID will build on that past work while striving to:

  • Enhance understanding of pathogen drug resistance and identify approaches to manage human infections by drug-resistant organisms;
  • Gain new insight into microbial diversity and evolution of pathogen populations and how these impact human infectious diseases;
  • Identify mechanisms and consequences of pathogen modulation of host response to infection, and understand how the pathogen interacts with host immune system and the host microbiome;
  • Characterize the genomic variation in and virulence of infectious diseases; and
  • Explore human immunity to malaria and influenza.

    GCID will be divided into research projects, each focused on viruses, bacteria and parasites.

Single-use bag for vaccines and drugs

Sartorius Stedim Biotech, France, is introducing a brand-new, scalable range of single-use bag family Flexsafe that enables the implementation of single-use bioprocessing throughout all steps of drug manufacture—all using just the one innovative polyethylene film. Flexsafe is based on a multilayer, proprietary polyethylene (PE) film, called S80. A standardized cell growth assay has been used to optimize film formulation, determine the operating ranges for extrusion, welding and gamma-irradiation processes and to establish specifications and process controls. Flexsafe ensures excellent and reproducible cell growth behavior of the most sensitive cell lines, guarantees lot-to-lot consistent cell growth performance. With its robust 400 μm thick PE film, Flexsafe is the strongest and most flexible bag currently on the market. The bags enable safe and easy-to-use operation in the most demanding applications such as liquid shipping and large-scale stirred bioreactors. Furthermore, batch-to-batch consistent extractables and leachables profiles support drug manufacturers throughout the entire lifecycle of modern biological treatments from clinical development to commercial supply many years after launch. Users can gain assurance that their initial extractable and leachable qualification work and data remain valid every time they operate their single-use Flexsafe bioprocess.

Commercializing colorectal cancer staging test in China

Shuwen Biotech Co. Ltd., China, has announced that it has formed an alliance with the Canada-based DiagnoCure, Inc. to commercialize the Previstage™ GCC colorectal cancer staging test in China. DiagnoCure has granted Shuwen Biotech an exclusive license to commercialize the Previstage™ GCC colorectal cancer staging test in the Greater China Region (China, Hong Kong, and Taiwan province of China). Shuwen Biotech will conduct clinical trials in China on an IVD kit format of the Previstage™ test, and obtain a regulatory approval from the Chinese FDA for the kit. Shuwen Biotech will manufacture the IVD kit in its own manufacturing facilities and market the kit to major Chinese hospitals. In addition, Shuwen Biotech will also offer the Previstage™ testing service from its own clinical reference lab.

Roche to buy U.S. biotech firm Seragon for up to $1.7 billion

Roche Holding AG ROG.VX would pay up to $1.725 billion to buy Seragon Pharmaceuticals, a privately-held U.S. biotech company that researches breast cancer treatments. San Diego-based Seragon was spun out from Aragon Pharmaceuticals last year when that company was bought by Johnson & Johnson JNJ.N. Seragon is focused on developing a new generation of oral medicines that it believes offer an improved way of tackling hormone receptor-positive breast cancer. Its most advanced experimental drug, ARN-810, is currently in initial Phase I clinical trials for breast cancer patients who have not responded to current hormonal agents. Roche said Seragon’s so-called oral selective estrogen receptor degraders, or SERDs, would complement existing research and development programmes in breast cancer under way at the Swiss group’s Genentech unit. The Basel-based drugmaker will pay $725 million in cash and may hand over as much as $1 billion more if Seragon achieves drug development milestones. Seragon is the second notable acquisition in as many months for Roche


‘Clever’ DNA may help bacteria survive

Scientists from Monash University, Australia, have discovered that bacteria can reshape their DNA to survive dehydration. The research shows that bacterial DNA can change from the regular double helix — known as B-DNA, to the more compact A-DNA form, when faced with hostile conditions such as dehydration. This unique process in DNA called the B-A-B transition allows it to change its structure in response to environmental change without impacting the ability of the bacteria to function and reproduce This finding may be important in understanding how dormant bacteria that are transferred from dry surfaces may become active and reproduce in the human body. Also, this change may have a biological function in bacteria, potentially assisting them to survive dehydration. The next phase of the research will see the team investigate how bacteria survive other conditions such as temperature, pH levels, oxygen, nutrients and antimicrobials and discover what role the ‘clever’ DNA plays under these conditions.

Scientists collaborate to sequence common bean’s genome

In the United States, University of Georgia researchers worked with a team of scientists to help sequence and analyze the genome of the common bean, Phaseolus vulgaris. Black beans, pinto beans, kidney beans, green beans, pole beans and others are varieties of the common bean. For the study, the team sequenced and assembled a 473-million basepair genome of the common bean. Thought to have originated in Mexico more than 100,000 years ago, the common bean was domesticated separately at two different geographic locations in Mesoamerica and the Andes. The team compared sequences from pooled populations representing these regions, finding only a small fraction of shared genes. This indicated that different events had been involved in the domestication process at each location. The team looked for regions associated with traits such as low diversity, flowering time and nitrogen metabolism. They found dense clusters of genes related to disease resistance within the chromosomes. They also identified a handful of genes involved in nitrogen fixing. These findings provide information on regions of the genome that have been intensely selected either during domestication or early improvement and thus provide targets for future crop improvement efforts.

Genome editing of crops may be restricted by EU

Genome editing typically involves finding the part of a plant genome that could be changed to render it less vulnerable to disease, or resistant to certain herbicides, or increase yields or other desirable traits. Researchers use “molecular scissors” to break apart the genome and repair it, which is a process that occurs naturally when plants are under attack from diseases and can throw up new mutations that enable the plant to survive future attacks. This fledgling technology is at risk of falling foul of the European Union’s genetic modification rules. According to European parliament’s Green party, there are many uncertainties as regards the impact of gene-edited organisms on the environment and health. Under EU laws, however, it is unclear whether gene editing should be treated in the same way as genetic modification. GM crops are effectively banned in Europe, and licences to experiment in GM are rare and very expensive. The European commission is expected to offer guidance on the technology soon. Lack of clarity on the legal status of gene editing techniques is hampering research and potential investment, particularly in Europe. The Green party/European Free Alliance group in the European parliament said that gene editing raises similar concerns as genetic modification as regards intellectual property rights and the impact on traditional and organic farming models and that it would make sense for gene editing to be covered by the same regulatory regime as existing GMOs.

Novel genomic changes in common type of lung cancer

Researcher from The Cancer Genome Atlas (TCGA) Research Network, the United States, has identified novel mutations in a well-known cancer-causing pathway in lung adenocarcinoma have been identified. This finding may expand the number of possible therapeutic targets for this disease and potentially identify a greater number of patients with treatable mutations because many potent cancer drugs that target these mutations already exist. These mutations put a cell signaling pathway known as the RTK/RAS/RAF pathway into overdrive. Mutations affecting the RTK/RAS/RAF pathway can cause it to become stuck in the “on” state. As a result, signals that promote cancer cell proliferation and survival are produced continuously. However, some drugs currently available curb aberrant activity of this pathway and prompt therapeutic responses in patients. Researchers identified gene mutations that would increase RTK/RAS/RAF pathway activity in 62% of the samples. The affected genes are oncogenes, or genes that have the potential to cause cancer when mutated or expressed at high levels. Consequently, these tumor samples were classified as oncogene-positive.

To identify additional alterations, the investigators looked at DNA copy number changes, or changes in gene number resulting from the deletion or amplification (multiplication) of sections of DNA in the genome. In doing so, they detected amplification of two oncogenes, ERBB2 and MET, which are part of the RTK/RAS/RAF pathway. Gene amplification usually leads to increased expression of the encoded protein in cells. Now that these amplifications have been identified, clinicians may be able to treat patients whose tumors have specific gene changes with drugs currently available or under development. Additional analysis identified other genes that may play important roles in lung cancer development. Mutations in one of these genes, NF1, had previously been reported in lung cancer; NF1 is a known tumor suppressor gene that regulates the RTK/RAS/RAF pathway. Mutations in NF1 also put the pathway into overdrive. Another mutated gene, RIT1, is also part of the RTK/RAS/RAF pathway, and this is the first study to associate mutation of this gene with lung cancer.

Blood test can reveal breast cancer risk

Scientists at the University College London, the United Kingdom, have identified an epigenetic signature in the blood of women who have a genetic mutation of the BRCA1 gene, which produces tumour-suppressing proteins. The mutation is inherited from a parent and the cause of at least 10% of breast cancer cases. But the researchers also found this epigenetic signature in women who don’t have a BRCA1 mutation, meaning the marker could help predict whether a woman would develop breast cancer several years before diagnosis.

Introducing synthetic features without genetic modification

A new study by researchers from the University of Trento, Italy, has shown that controlling organisms on the cellular level does not necessarily require genetic modification. Escherichia coli (E. coli) behavior can be controlled by constructing artificial cells that first sense molecules that E. coli alone cannot sense, and then release different molecules that E. coli can sense. In a way, the artificial cells act as translators by converting unrecognized signals into a chemical language that organisms can understand. The translated signal can then potentially trigger a controllable response in the organism. In their experiments, the researchers constructed artificial cells that contain a special vesicle which in turn contains several biological components, including a chemical that E. coli can sense (isopropyl b-D-1 thiogalactopyranoside, or IPTG) and DNA that encodes for a riboswitch that responds to an external stimulus. In this case, the external stimulus is the molecule theophylline, commonly found in cocoa beans. When the artificial cell’s riboswitch detects the presence of theophylline, it activates the translation process: a small pore opens in the cell, resulting in the release of IPTG. The E. coli responds to IPTG by exhibiting a green fluorescence. Although E. coli does not respond to theophylline on its own, the artificial cells effectively “expand the senses” of the bacteria by allowing it to indirectly respond to theophylline by translating the chemical message. In this way, E. coli’s cellular behavior can be controlled without the need for genetic engineering.

The new strategy can potentially overcome the disadvantages of genetic engineering, including the technical difficulties and unintended side effects. One application is using bacteria to search for and clean up environmental contaminants. Instead of genetically engineering bacteria to do this, artificial cells could be constructed to sense the contaminant molecules and release chemoattractants that lure natural bacteria capable of feeding on the contaminants to the site. Artificial cells could also be used for medical applications, such as to destroy tumors and bacterial infections. For example, rather than spraying engineered bacteria into the lungs of cystic fibrosis patients, artificial cells could be built to detect the presence of specific biofilms, and then release small molecules to disperse the biofilms and thus clear the infection. Similar strategies could also be used to replace engineered probiotics in food and supplements with artificial cells that communicate with gut microbiota to prevent disease.


Self-assembling protein nanomachines

The ability to design new protein nanostructures could have useful implications in targeted delivery of drugs, in vaccine development and in plasmonics. A recently developed computational method Rosetta macromolecular modeling package may be an important step toward that goal. The project was led by the University of Washington’s Neil King, translational investigator; Jacob Bale, graduate student in Molecular and Cellular Biology; and William Sheffler in David Baker’s laboratory at the University of Washington Institute for Protein Design, the United States, in collaboration with colleagues at UCLA and Janelia Farm. With the new software the scientists were able to create five novel, 24-subunit cage-like protein nanomaterials. The method involved encoding pairs of protein amino acid sequences with the information needed to direct molecular assembly through protein-protein interfaces. The interfaces not only provide the energetic forces that drive the assembly process, they also precisely orient the pairs of protein building blocks with the geometry required to yield the desired cage-like symmetric architectures.

Creating this cage-shaped protein may be a first step towards building nano-scale containers. We can look forward to a time when cancer-drug molecules will be packaged inside of designed nanocages and delivered directly to tumor cells, sparing healthy cells. Packaging the drugs inside customized nanovehicles with parking options restricted to cancer sites might circumvent the side effects. Building nano-decoys may be a way to train the immune system to attack certain types of pathogens.

Protein discovery may lead to cure for hearing loss

A scientific team led by Christine Petit, professor at the Institut Pasteur in Paris and at Collège de France, France, reports that the absence of a specific protein in the inner ear or impairment of the gene that codes for it leads to profound deafness in mice and humans. The researchers believe that it is possible to develop gene therapy strategies for deafness caused by defects in this gene. Three types of protocadherin-15 are known to exist in auditory sensory cells of the inner ear and the CD2 isoform of protocadherin-15 is an essential component of the tip-link and the absence of protocadherin-15 CD2 in mouse hair cells results in profound deafness.

The researchers engineered mice that lack only the CD2 isoform of protocadherin-15 exclusively during adulthood. While the absence of this isoform led to deafness, the lack of the other protocadherin-15 isoforms in mice did not affect their hearing. The scientists point out that these results provide critical information for the identification of new components of the mature auditory mechano-electrical transduction machinery. This will also serve as a basis for the development of gene therapy for deafness caused by PCDH15 defects.

Human protein cleans bacteria from drinking water

Strains of Escherichia coli such as enterohemorrhagic E. coli O157 are life-threatening to humans. Fruit and vegetables washed in contaminated water can cause severe food-poisoning and even death. Teruyuki Komatsu and co-workers at Chuo University, Tokyo, Japan, have shown that they can remove E. coli from drinking water using tiny tubes made of human serum albumin. This method begins by depositing microtubes made from alternating layers of human serum albumin (HSA) and poly-L-arginine onto a polycarbonate template. The template is then dissolved to leave a hollow tube, which is just the right size to fit the E. coli. Key to removing E. coli from a solution is its strong binding affinity for HSA, which attracts the bacteria into the tube. So effective is this binding, that just 1.5μg of microtubes, added to a liter of contaminated water containing 100,000 bacteria, was able to remove the bacteria with almost 100% efficiency. Finally, a layer of magnetite (iron (II) oxide) nanoparticles is incorporated into the microtubes to allow their easy removal from the solution using a magnetic field. ‘Because the microtubes are made mainly from human protein, they are safe for medical use.

Dyes used to paint new picture of disease

In the United States, George Mason University researchers unraveled the mystery of deciphering the contact points where proteins touch each other. One protein interlocks with another protein like adjacent pieces in a jigsaw puzzle, and this sends a signal down the line to the next protein. The mystery is in the “hot spots” where proteins interlock. Researchers know which proteins connect but couldn’t pinpoint where it happens. Dyes—the type used in common copying machines and textiles—are mixed with proteins. The dye paints the proteins everywhere except where the proteins are connected to one another. Then the proteins are disconnected but the dye remains, excluding the blank spot where the proteins were “kissing.” Finding ways to break up interlocking proteins could be used to locate new drug targets. Pharmaceutical companies could use the Mason-developed process to create drugs that break up the protein-to-protein connection or stop it from happening altogether.

Engineering a protein to prevent brain damage from toxic agents

Research at New York University, the United States, is paving the way for a breakthrough that may prevent brain damage in civilians and military troops exposed to poisonous chemicals – particularly those in pesticides and chemical weapons. Phosphotriesterases have the unique capability of degrading chemicals in a class known as organophosphates. Organophosphates permanently bond to neurotransmitters in the brain, interfering with its ability to function and causing irreversible damage.

In a process that married computational biology and experimentation, the collaborators used Rosetta computational modeling software to identify sequences in the fluorinated phosphotriesterase protein that could be modified to increase its stability and make therapeutic applications a reality. The possibilities for this reengineered protein are considerable. In addition to therapeutic formulations, which could prevent nerve damage in the event of a gas attack or pesticide exposure, the proteins could be critical when stores of toxic nerve agents need to be decommissioned.

HSP critical in malaria parasite protein trafficking

The mechanism by which P. falciparum exports hundreds of proteins into the host erythrocyte in order to mediate its survival and virulence has been poorly understood. Researchers on the topic have suggested the existence of a Plasmodium translocon of exported proteins (PTEX) to account for this activity. A translocon is a protein complex in the plasma membrane that governs protein secretion and membrane protein insertion. The translocon channel provides a route for proteins to pass through the hydrophobic barrier of the membrane Investigators at Washington University school of medicine looked at the possible involvement of the heat shock protein HSP101 in this process. By helping to stabilize partially unfolded proteins, HSPs aid in transporting proteins across membranes within the cell.

Inhibiting HSP101 function results in a nearly complete block in protein export from the parasitophorous vacuole with substrates accumulating in the vacuole in both asexual and sexual parasites. This block extended to all classes of exported proteins, revealing HSP101-dependent translocation across the PVM as a focal point in a multi-pathway export process. P. falciparum that lacked the gene for HSP101 or another suggested PTEX protein, PTEX150, demonstrated greatly reduced trafficking of all classes of exported proteins beyond the double membrane barrier enveloping the parasite. Moreover, the export of proteins destined for expression on the infected erythrocyte surface, including the major virulence factor PfEMP1 in P. falciparum, was significantly reduced in the PTEX knockdown parasites. PTEX function was also found to be essential for blood-stage growth, as even a modest knockdown of PTEX components had a strong effect on the parasite’s capacity to complete the erythrocytic cycle both in vitro and in vivo.


Virus that kills cancer and tumour cells

According to Penn State College of Medicine researchers in the United States, a virus can be used to kill triple-negative breast cancer cells and tumours grown from these cells in mice, finds a new research. Understanding how the virus kills cancer cells may lead to new treatments for breast cancer Adeno-associated virus type 2 (AAV2) infects humans, but is not known to cause sickness. The researchers tested AAV2 on a cell-line representative of triple-negative breast cancer. The AAV2 killed 100% of the cells in the laboratory by activating proteins called caspases, which are essential for the cell’s natural death. AAV2 mediated cell killing of multiple breast cancer cell lines representing both low and high grades of cancer and targeted the cancer cells independent of hormone or growth factor classification. These results are significant, since tumour death in response to therapy is also used as the measure of an effective chemotherapeutic.

Scientists regrow human corneas from adult stem cells

Researchers from the Massachusetts Eye and Ear Research Institute, the United States, have identified a way to enhance regrowth of human corneal tissue to restore vision, using a molecule known as ABCB5 that acts as a marker for hard-to-find limbal stem cells. Limbal stem cells — identified with a new marker — could reverse a leading cause of blindness. Limbal stem cells reside in the eye’s basal limbal epithelium, or limbus, and help maintain and regenerate corneal tissue. Their loss due to injury or disease is one of the leading causes of blindness. Researchers were able to use antibodies detecting ABCB5 to zero in on the stem cells in tissue from deceased human donors and use them to regrow anatomically correct, fully functional human corneas in mice. This finding will now make it much easier to restore the corneal surface. It’s a very good example of basic research moving quickly to a translational application,” said Ksander.

Anti-diabetic drug slows aging and lengthens lifespan

A study conducted by researchers from KU Leuven and Ghent University, Belgium, has provided evidence that metformin, the world’s most widely used anti-diabetic drug, slows aging and increases lifespan. The drug causes an increase in the number of toxic oxygen molecules released in the cell and this, surprisingly, increases cell robustness and longevity in the long term. Mitochondria generate tiny electric currents to provide the body’s cells with energy. Highly reactive oxygen molecules are produced as a by-product of this process. While these molecules are harmful because they can damage proteins and DNA and disrupt normal cell functioning, a small dose can actually do the cell good. Metformin causes a slight increase in the number of harmful oxygen molecules. This makes cells stronger and extends their healthy lifespan. The researchers studied metformin’s mechanism in the tiny roundworm Caenorhabditis elegans, an ideal species for studying aging because it has a lifespan of only three weeks. Worms treated with metformin show very limited size loss and no wrinkling. They not only age slower, but they also stay healthier longer.

Scientists make human blood using stem cells

A team of researchers from University of Wisconsin – Madison, the United States, have cracked the elusive code to turn stem cells into human blood. They discovered two genetic programs responsible for taking stem cells and turning them into both red and white cells that make up human blood. For every million stem cells the researchers were able to produce 30 million blood cells. The finding is important as it identifies how nature itself makes blood products at the earliest stages of development. The latest breakthrough identifies two distinct groups of transcription factors that can directly convert human stem cells into the hemogenic endothelial cells, which subsequently develop into various types of blood cells.

Method for controlling antibiotic-resistant bacteria

The World Health Organization (WHO) has warned that antibiotic resistance in bacteria is spreading globally, causing severe consequences. And even common infections that have been treatable for decades can once again kill. Researchers from the University of East Anglia, the United Kingdom, investigated gram-negative bacteria, which are particularly resistant to antibiotics because of its cells’ impermeable lipid-based outer membrane. The new findings reveal how bacterial cells transport the barrier building blocks (lipopolysaccharides) to the outer surface. The structure, molecular dynamics simulations, and functional assays suggest that the hydrophilic O-antigen and the core oligosaccharide of the LPS may pass through the barrel, and the lipid A of the LPS may be inserted into the outer leaflet of the outer membrane through a lateral opening between strands β1 and β26 of LptD. These findings not only help us to understand important aspects of bacterial outer membrane biogenesis, but also have significant potential for the development of novel drugs against multi-drug resistant pathogenic bacteria.

Genetically modified mosquitoes offer hope in malaria fight

Scientists from Imperial College London, the United Kingdom, have found a way of genetically modifying mosquitoes to produce sperm that only creates males, offering a potential fresh approach to fighting and eventually eradicating malaria. Researchers tested a genetic method that distorts the sex ratio of Anopheles gambiae mosquitoes, so that the female mosquitoes that bite and pass the disease to humans are no longer produced. The team reported that in the first laboratory tests, the technique created a fully fertile mosquito strain that produced 95% male offspring. In their experiments, the scientists inserted a DNA cutting enzyme called I-PpoI into Anopheles gambiae mosquitoes. The enzyme works by cutting the DNA of the female-producing X chromosome during production of sperm, so that almost no functioning sperm carry it. As a result, the offspring of the genetically modified mosquitoes was almost exclusively male. The scientists introduced the genetically modified mosquitoes to five caged wild-type mosquito populations. In four of the five cages, this eliminated the entire population within six generations due to the lack of females. The hope is that if this could be replicated in the wild, this would ultimately cause the malaria-carrying mosquito population to crash.

Human stem cells successfully transplanted, grown in pigs

The rejection of transplants and grafts by host bodies is a huge hurdle for medical researchers. A study has shown that a new line of genetically modified pigs will host transplanted cells without the risk of rejection. A team of researchers from University of Missouri, the United States, implanted human pluripotent stem cells in a special line of pigs developed with immune systems that allow the pigs to accept all transplants or grafts without rejection. Once the scientists implanted the cells, the pigs did not reject the stem cells and the cells thrived. Achieving this success with pigs is notable because pigs are anatomically similar to humans than other animals. This means that research in pigs is more likely to have results similar to those in humans for many different tests and treatments.

Light-sensing retina is created using stem cells

A section of light-sensing tissue, closely resembling the human retina, has been grown in the laboratory from human stem cells at John Hopkins University, the United States. The retinas were grown using human induced pluripotent stem cells (iPSCs), adult cells that have been genetically reprogrammed to behave similarly to embryonic stem cells. Using iPSCs, the researchers first generated precursor retinal cells by growing them in a medium containing specific growth factor chemicals. The cells assembled into a complex structure, in steps similar to those observed in the eye during fetal development. The tissue not only structurally resembled the human retina, but it also contained all major retinal cell types arranged in their proper layers. Scientists found that their artificial retina contained cells that not only resembled photoreceptors structurally, but were also able to respond to light. This finding advances opportunities for vision-saving research and may ultimately lead to technologies that restore vision in people with retinal diseases. This light-sensing only provides a small part of the vision-making process as lab retina is not capable of producing a visual signal that the brain can interpret into an image.

Supercooling organs will help save lives

A new supercooling technique could increase the amount of time human organs can remain viable for transplant, it could help save more lives and reduce wastage of donated organs. Researchers from the Center for Engineering in Medicine at Massachusetts General Hospital (MGH), the United States, have developed a new supercooling technique to increase the amount of time human organs could remain viable outside the body. This study was conducted in rats, and if it succeeds in humans, it would enable a world-wide allocation of donor organs, saving more lives.

The first step is to employ the use of machine perfusion—a way of delivering oxygen and nutrients to capillaries in biological tissues while outside the body—to supercool the liver tissue without causing irreversible damage to the cells. In order to accomplish this, 3-OMG (3-O-methyl-D-glucose), a non-toxic, glucose compound, was added to the solution being delivered to the liver. The 3-OMG is taken up and because it cannot be metabolised by cells, accumulates in the liver cells, acting as a protectant against the cold. The solution was modified by adding PEG-35kD (polyethylene glycol) to specifically protect cell membranes. Ethylene glycol is the active ingredient in anti-freeze, and it works by lowering the freezing point of a solution. The livers were then slowly cooled below the freezing point, to 21 degrees Fahrenheit, without inducing freezing—thereby supercooling the organ for preservation. After storing the organs for several days, the researchers again used machine perfusion to rewarm the organ, while also delivering oxygen and other nutrients to prepare the organ for transplantation.

Using the technique, they were able to store the rat livers for three days and four days at 21 degrees Fahrenheit. The survival rate for animals receiving livers stored for four days was 58%. The next step will be to conduct similar studies in larger animals. But the technique’s achievement in being the first method to have a successful survival rate after the livers had been stored for three days and possible potential for four-day storage has broad implications for the future.

Contraceptive implant

MicroCHIPS, the United States, has developed a contraceptive implant that can be deactivated and reactivated via wireless remote. A woman, with a remote control, could turn off the implant or with another click of the remote could restart it. Device measures 20 x 20 x 7 mm, implanted under the skin of the buttocks, upper arm, or abdomen. The implant is designed to last up to 16 years. The device is designed to dispense 30 µg a day of levonorgestrel. The latter is a progestin used as an ingredient in some hormonal contraceptives. Sixteen years’ worth of the hormone fits in tiny reservoirs on a microchip 1.5 cm wide inside the device. MicroCHIPS has a hermetic titanium and platinum seal on the reservoirs containing the levonorgestrel. Passing an electric current through the seal from an internal battery melts it temporarily, allowing a small dose of the hormone out each day. After 16 years, the device could be removed. The goal is to have it on the market by 2018.

‘Nanojuice’ could improve how doctors examine the gut

In the United States, University of Buffalo researchers are developing a new imaging technique involving nanoparticles suspended in liquid to form “nanojuice” that patients would drink. Upon reaching the small intestine, doctors would strike the nanoparticles with a harmless laser light, providing an unparalleled, non-invasive, real-time view of the organ. The advancement could help doctors better identify, understand and treat gastrointestinal ailments.

In laboratory experiments performed with mice, the researchers administered the nanojuice orally. They then used photoacoustic tomography (PAT), which is pulsed laser lights that generate pressure waves that, when measured, provide a real-time and more nuanced view of the small intestine. The researchers plan to continue to refine the technique for human trials, and move into other areas of the gastrointestinal tract.


Genetic hierarchy in plant sperm cell formation undressed

A team of biologists from the University of Leicester, the United Kingdom, has solved a mystery surrounding how plants have sex The researchers have discovered a pair of proteins made by flowering plants that are vital for the production of the sperm present within each pollen grain. Flowering plants require not one, but two sperm cells for successful fertilisation: one to join with the egg cell to produce the embryo and one to join with a second cell to produce the nutrient-rich endosperm inside the seed. The mystery of this ‘double fertilization’ process is how each single pollen grain is able to produce twin sperm cells. This study has found a pair of genes called DAZ1 and DAZ2 that are essential for making twin sperm cells. Plants with mutated versions of DAZ1 and DAZ2 produce pollen grains with a single sperm that is unable to fertilize.

The researchers show that DAZ1 and DAZ2 are controlled by the protein DUO1 that acts as a ‘master switch’ -- so that DUO1 and the DAZ1/DAZ2 genes work in tandem to control a gene network that ensures a pair of fertile sperm is made inside each pollen grain. Interestingly, DAZ1 and DAZ2 perform their role by cooperating with a well-known ‘repressor’ protein called TOPLESS that acts as a brake on unwanted gene activity that would otherwise halt sperm and seed production. Although TOPLESS has many roles in plants it has not previously been linked sperm production. Given their important role in male fertility, the discovery of DAZ1 and DAZ2 has the potential to be applied in the development of new plant breeding techniques to prevent the unwanted passing of genes – or ‘horizontal gene transfer’ – between crops or from crops to wild species. This new knowledge also generates genetic tools and new ways of thinking about, and monitoring the effects of environmental stresses on the reproductive process.

“Seed” enhancers seize control during stem cell development

By examining the shifts in gene transcription that occur when stem cells progress from the embryonic to the epiblast stage, researchers at Case Western Reserve, the United States, have identified a new class of genetic switch. Using transcriptomic and epigenomic mapping, the researchers showed that a small fraction of transcripts are differentially expressed between mouse embryonic stem cells (mESCs) and primed mouse epiblast stem cells (mEpiSCs). These genes show expected changes in chromatin at their promoters and enhancers. In mESCs, these genes are associated with dominant proximal enhancers and dormant distal enhancers. In mEpiSCs, the naive-dominant enhancers are lost, and the seed enhancers take up primary transcriptional control. Unlike most enhancers, which are only active in specific times or places in the body, seed enhancers play roles from before birth to adulthood. They are present, but dormant, in the early mouse embryonic stem cell population. In the more developed mouse epiblast stem cell population, they become the primary enhancers of their associated genes.”

Super bananas – world’s first human trial

The world’s first human trial of pro-vitamin A-enriched banana will start soon. The QUT project is one of the most significant biofortification projects in the world today. The bananas have been harvested from the QUT field trial in Innisfail, north Queensland and transported to the United States for the world-first human trial. The human trial will last for six weeks with conclusive results known by the end of the year. While on the outside the banana plants looked just like any other banana the inside was a whole other story. The banana flesh of a pro-vitamin A-enriched banana is orange rather than the cream colour we are used to and in fact the greater the pro-vitamin A content the more orange the banana flesh becomes.

Getting a jump on plant-fungal interactions

Transposable elements (TEs) are DNA sequences that can “jump” around the genome that were originally dismissed as junk DNA. As more genome sequences have become accessible, researchers are finding that transposable elements play roles in genomic variation. Fungal researchers have found a correlation between symbiotic plant pathogens and the amount of transposable elements.

Working with collaborators at Harvard University, the DOE JGI has already sequenced and assembled the genome of Amanita thiersii, a fungus commonly found on lawns that decomposes leaf litter. For this project, the genome of A. thiersii and of A. muscaria, another fungus sequenced and assembled at the DOE JGI, were compared against other fungal genomes that came from samples collected around the world.

The team compiled an aggregate TE library from all those found in the fungal genomes used in the analysis. Among their findings was that there are significantly more TEs in the genomes of ectomycorrhizal fungi compared to asymbiotic fungi. A. thiersii’s genome had more TEs relative to the amount of coding sequence than any of the other fungal genomes in the analysis. Additionally, in the ectomycorrhizal fungi, some TEs were “younger” and had been inserted later.

‘Most famous wheat gene’ discovered

In the United States, Washington State University researchers have found ‘the most famous wheat gene’. The discovery clears the way for breeders to develop wheat varieties with the disease- and pest-resistance traits of other grasses. Their first effort involves transferring a gene from jointed goatgrass, a wild relative of wheat, to confer resistance to stripe rust. The fungus is considered the world’s most economically damaging wheat pathogen. While facilitated by technology, the actual exchange of genetic material is similar to what has long taken place in nature, only faster. Incorporating the gene transfer into the overall breeding process, researchers can develop a new variety in five years.

Boron facilitates stem cell growth, development in corn

Boron deficiency is one of the most widespread causes of reduced crop yield. Researchers at the University of Missouri, the United States, have found that boron plays an integral role in development and reproduction in corn plants. Scientists anticipate that understanding how corn uses the nutrient can help farmers make informed decisions in boron-deficient areas and improve crop yields. Researches found that boron deficiency causes a problem in the meristems, or the stem cells of the plant, which was unknown before.

Through a series of experiments involving scientists from several disciplines at University of Missouri, the scientists were able to piece together the puzzle and reach a new conclusion.” The research evaluated a group of plants stunted by its ability to grow tassels. Kim Phillips, a graduate student in McSteen’s lab, mapped the corn plant’s genome and found that a genetic mutation stunted tassel growth because it was unable to transport boron across the plant membranes, inhibiting further growth in the plants. Further testing revealed that, at the cellular level, the affected plants’ meristems had altered pectin which is strengthened with boron and stabilizes the plant cell. Without the pectin, plant meristems disintegrate.

Researchers discover rust-resistance genes in sunflower

Two genes that protect sunflowers against rust disease have been discovered by U.S. Department of Agriculture (USDA) scientists.Agricultural Research Service (ARS) molecular geneticist Lili Qi at the agency’s Sunflower and Plant Biology Research Unit in Fargo, North Dakota, and her collaborators discovered that the genes, R13a and R13b, confer resistance against all rust strains tested to date. Her collaborators include Thomas Gulya and Brent Hulke at the ARS Fargo unit, and Li Gong and Samuel Markell with North Dakota State University. ARS is USDA’s chief intramural scientific research agency, and this research supports the USDA priority of promoting international food security.

The R13a gene was found in the confection sunflower line called HA-R6, while the R13b gene was in the oilseed line RHA 397. The USDA inbred line HA-R6 is one of the few confection sunflower lines resistant to rust.

Kernels infected by rust can be damaged and discolored and are therefore unlikely to meet grading standards established by the industry for confection sunflower seeds.The rust resistant lines should be very useful to breeders who want to develop rust-resistant commercial sunflower hybrids.The research was published in Theoretical and Applied Genetics.


The future of Indian agriculture

Locating India’s resource endowments in the context of its fast growing economy, the book assesses the growing demands on the country’s agricultural sector. In doing so, it discusses the distinctive and rapidly diversifying food needs in rural and urban areas, the supply potential of India’s agricultural resources and the impact of technology. Modelling an alternative growth path, the book offers policy solutions to the challenges of inflation, poverty and food security. Written by one of India’s foremost agricultural economists, the book offers incisive insights on die future of Indian agriculture.

Contact: National Book Trust, 5, Nehru Bhawan, Institutional Area, Phase-II Vasant Kunj, New Delhi -110070 Tel: 011-26707700; E-mail:

Lewin’s Genes XI

Molecular biology is a rapidly advancing field with a constant flow of new information and cutting-edge developments that impact our lives. Lewin’s GENES has long been the essential resource for providing the teaching community with the most modern presentation to this dynamic area of study. GENES XI continues this tradition by introducing the most current data from the field, covering gene structure, sequencing, organization, and expression.

Contact: Jones & Bartlett Learning, 5 Wall Street, Burlington, MA 01803, Tel:(800) 832-0034, (978) 443-5000 (Switchboard); Customer service: ext. 8197, Fax:(978) 443-8000

Lipid Nanoparticles: Production, Characterization and Stability

This work sets out to provide the reader with a clear and understandable understanding of the current practices in formulation, characterization and drug delivery of lipid nanoparticles. A comprehensive description of the current understanding of synthesis, characterization, stability optimization and drug incorporation of solid lipid nanoparticles is provided. The work provides a detailed description of the types of lipid nanoparticles available (e.g. SLN, NLC, LDC, PLN) and how they range from imperfect crystalline to amorphous in structure. Current thoughts on where drugs are situated (e.g. in the core, or at the interface) and how this can be manipulated are discussed. The many techniques for production, including the author’s own variant of microwave heating, are fully discussed.

Contact: Springer Science+Business Media Singapore Private Limited, 152 Beach Road, #22-06/08 Gateway East, Singapore 189721, Singapore; E-mail:


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