Some of our most popular processed foods contain small amounts of toxic acrylamide. Currently available methods that lower the accumulation of this reactive compound have a negative effect on sensory characteristics and/or are not broadly applicable. Realizing their limited options, several food companies have committed to substantially reducing the acrylamide levels in fried and baked potato products over the next three years. A novel method that could be applied to produce the desired low-acrylamide foods was recently published in Plant Biotechnology Journal. According to this method, potato plants are transformed with an all-native silencing construct that targets two asparagine synthetase genes. The resulting plants produce tubers with very low levels of the acrylamide precursor asparagine. French fries and potato chips from these "intragenic" plants contain up to 20-fold lower levels of acrylamide than their untransformed counterparts. Given the important role of processed potato products in the modern Western diet, a replacement of current varieties by the low-asparagine potatoes would reduce the average daily intake of acrylamide by almost one-third.

Acrylamide is a typical constituent of modern diets and may play a minor role in the emergence of certain "modern" diseases. Dietary intake levels of acrylamide have been rising in the Western world since the early 1900s and are currently estimated at 40 µg/person/day. Acrylamide is formed in starchy foods when they are baked or fried as a consequence of the reaction between certain sugars and the free amino acid asparagine (Fig. 1). This reactive compound is present in products derived from wheat flour, coffee beans, and potato tubers. Ingested acrylamide is readily absorbed and, in part, metabolized by a cytochrome P450 to produce mercapturic acid and glycidamide. Whereas mercapturic acid is excreted via urine, both the remaining acrylamide and its reactive metabolite bind to various proteins as well as DNA. High levels of adduct formation have been linked to animal health issues, including cumulative nerve terminal damage. As suggested by several recent studies, acrylamide intake may also be associated with the increased incidence of neurodegenerative and other types of diseases^1-3.

Water availability is the primary limiting factor of global crop yields. Consequently, considerable effort is devoted to converging breeding and technological research to develop crops with improved performance under water-limiting conditions. Transgenic plants tolerant to abiotic stress are being developed through the introduction of multiple genes; however, successfully producing stress tolerant crops remains a challenging task.

Tolerance to abiotic stresses is improved by the expression of bacterial cold shock proteins (CSP). A small set of E. coli cold shock proteins (CSPs) exhibits a prototypical cold shock domain (CSD) during cold treatment. Expression of related cold shock proteins from bacteria—CspA from Escherichia coli and CspB from Bacillus subtilis—promotes stress adaptation in multiple plant species.

Both CSPs and CSDs are found in bacteria and eukaryotes, including plants. In bacteria, CSD proteins (7 – 10 kD) contain sufficient nucleic acid binding activity to function as RNA chaperones. RNA chaperones—often called "protein chaperones"—are proteins that aid RNA folding by preventing or resolving misfolded species of RNA, in contrast to proteins that assist protein or RNA folding by catalyzing steps along the folding pathway or by stabilizing the final folded protein or RNA structure. RNA chaperones in bacteria favor active transcription, translation, and/or ribosome assembly. Research on CSPs and CSDs confirms that the endogenous function of CSPs in plants depends on RNA binding/chaperone activity through the CSD, and CSPs regulate stress responses through a post-transcriptional mechanism.

Recently, Monsanto Company researchers showed that bacterial CSPs can confer improved stress adaptation in multiple plant species by demonstrating improved stress tolerance in both E. coli and maize, and reported further 'proof of concept' in dicots and monocots—Arabidopsis, rice, and maize. Expression of bacterial CSPs improves cold tolerance in transgenic Arabidopsis when compared to nontransgenic controls. Similar experiments expressing CspA and CspB in transgenic rice show improved plant growth rates in plants exhibiting an increased tolerance to a number of abiotic stresses like cold, heat, and water deficit.

Further abiotic stress testing of transgenic CspA and CspB in maize demonstrates that transgenic expression by CspB improves vegetative growth performance. Twenty-two CspB transgenic events were evaluated under water limited field trials using commercial grade corn in an environment that received no rainfall during the target period. The best performing events show a growth rate increase of 12% and 24% under water deficit conditions. The CspB-expressing plants also demonstrate significant improvements in chlorophyll content by 2.5%, increasing the photosynthetic rates by 3.6% across all events. Transgenic maize plants expressing CspA under greenhouse conditions were also tested.

Increases in plant growth rates, chlorophyll content, and photosynthetic efficiency are key indicators of plant productivity and are expected to improve the overall yield of the plant. Therefore, the reproductive performance of CspB-expressing maize plants was also evaluated by harvesting all kernel-bearing ears from six replicates (34 plants per replicate) for each of six events selected for harvest, based on the magnitude of their improved vegetative performance. An across-event analysis demonstrates significant improvements were made in the number of plants showing an increased number of kernels per plant. These improvements are congruent with the expected results based on the timing of limited-water treatment, which was provided during the late vegetative stages and early immature ear development, and was relieved with sufficient water during the pollination and grain-fill periods.

Grain yield trials were also carried out under water deficit stress and non-stress conditions on 10 CspA- and 10 CspB-positive events that had demonstrated superiority in vegetative performance. Grain yield data were collected from four field sites where water was limited during the late vegetative phase of development, a treatment similar to the initial water deficit trial. An across-event analysis demonstrates that CspA transgenic lines contribute to a yield increase of 4.6% under water stress, with the two best performing events showing 30.8% and 18.3% improvement. CspB transgenic plants show 7.5% improved yield averages over controls; the best two performing events, CspB-Zm events 1 and 2, demonstrate yield improvements of 20.4% and 10.9%, respectively. These are the same two events that demonstrate significant improvements in leaf growth, chlorophyll content, and photosynthetic rates, indicating that these improvements in vegetative productivity translate into improvements in reproductive performance and grain yield.

To further investigate the ability of the CspB gene to provide tolerance to maize under water deficit conditions, CspB-Zm event 1 was deployed into three hybrid backgrounds and evaluated under two distinct stress treatment conditions at five replicated locations. The two treatments result in a decrease of overall yield of approximately 50%, relative to well-watered treatments. When compared to controls, the CspB transgenics demonstrate improved yields by at least 0.5 t/ha across 12 out of 15 reproductive stress treatments. The multi-year analysis with CspB-Zm event 1 shows the stability of yield advantages across locations under water-limited conditions, which proves the utility of this technology across the US maize growing regions.

The performance of CspB-Zm event 1 was also assessed by combining yield performance data from three hybrid test crosses collected over four years. The event yielded a 10.5% average benefit across the four years. CspB-Zm event 1 was also tested under western dryland maize conditions without supplemental water. Under these conditions when compared to the non-transgenic control, the CspB transgenic event contributed a yield benefit of up to 0.75 t/ha or 15%.

In summary, constitutive expression of E. coli CspA and bacterial RNA chaperones can confer abiotic stress tolerance in transgenic Arabidopisis, rice, and maize. This technology also provides a stable yield improvement under water limiting conditions.

Patent litigation can spawn a verdict of inequitable conduct. It may not sound that bad. Yet the ruling brands a patent unenforceable.

The verdict typically comes about like this. The defendant company in a patent infringement lawsuit alleges that the plaintiff company committed inequitable conduct during patent prosecution, the process of obtaining a patent. Inequitable conduct includes misrepresentation of a material fact, failure to disclose material information, or submission of false material information. Materiality is defined by what a reasonable examiner would have considered important in deciding whether to allow a patent application. The prohibited acts are coupled with intent to deceive, an intent that a judge may infer from the facts. If the judge decides that the plaintiff used deceit to obtain the patent, then the judge will issue the inequitable conduct verdict. It no longer matters that the defendant infringed, because courts will no longer enforce the patent.

An unenforceable patent still has uses – hiding unsightly cracks in a wall, for instance. An inventor who played a role in events leading to the patent-killing verdict may find fractures in his or her reputation. So, it's best to avoid those activities identified by courts as meriting the inequitable conduct death knell.

Submitting prior art can create hazards as well. Semiconductor Energy Laboratory learned this lesson the hard way when the company submitted an untranslated 29-page Japanese reference with a concise explanation of its relevance and a one-page partial English translation. District and appellate judges concluded that untranslated portions of the reference contained information more relevant to Semiconductor's claimed invention than anything else considered by the patent examiner.

Patent prosecution can take a long time, particularly in the field of biotechnology. As one patent application undergoes examination, a related application may issue as a patent and become the subject of litigation. Patent applicants should not only update their examiners about prior art that an opposing party offers in court, but also keep in mind that they may need to inform a patent examiner about a ruling to narrow claim scope. In Mallinckrodt, Inc. v. Masimo Corp., Masimo had not notified its patent examiner about a judge's decision that claims in a related patent must include certain limitations to avoid prior art.

The US patent office can assign two related applications to different patent examiners. When one examiner finds new prior art, the applicant may need to notify the other examiner about the discovery. In McKesson Information Solutions, Inc. v. Bridge Medical, Inc., the patent applicant had similar applications under review by two examiners. One examiner brought a newly-discovered prior art patent to the attention of McKesson. The company's failure to inform the other examiner about the prior art inspired the ruling of inequitable conduct.

Patent applicants can violate their duty of good faith by withholding information material to a determination about patentability. Merck lost a patent on a method of using cyclobenzaprine to treat certain types of skeletal muscle disorders, because a court found several examples of inequitable conduct. In one instance, Merck told the patent examiner that cyclobenzaprine did not cause drowsiness, even though the company had data showing that this assertion was untrue. In a dispute over Taq enzyme patent claims, a district court judge concluded that Cetus Corporation inventors withheld data that would have contradicted their arguments for patentability.

Information deemed material to patent examination can take many forms, as shown by Monsanto v. Bayer Bioscience. Here, the Court of Appeals for the Federal Circuit agreed with a district court's finding of inequitable conduct and the unenforceability of four Bayer patents. Plant Genetic Systems, a predecessor of Bayer Bioscience, had filed patent applications on Agrobacterium tumefaciens techniques for transforming plant cells with a fragment of a Bacillus thuringiensis toxin gene. During patent prosecution, the applicant disclosed an abstract of a poster presented at a conference by a scientist from a different organization. The patent applicant argued that the abstract did not render the claims obvious, because it failed to show that chimeric Bt toxin genes could be introduced into plant cells and would direct the synthesis of toxin. However, one of the patent applicant's employees had attended the conference and taken notes about the poster. The notes revealed details on the construction of a Bt toxin chimeric gene and expression vector, evidence that the Bt toxin fusion protein functioned as an insecticide, and an indication that the scientist had transformed a plant with a chimeric gene construct. Although the notes had been circulated among company officials and employees, the company had not submitted the notes to the patent examiner.

An accurate identification of inventors is vital to the health of a US patent. While the patent office allows patent applicants to correct honest mistakes about inventorship, courts view a deliberate misrepresentation of inventorship to constitute inequitable conduct. PerSeptive Biosystems, Inc. lost patents when a court decided that its inventors – with deceptive intent – had failed to name research collaborators from another company as co-inventors.

The Danger of Misleading Statements
Patent applications may include "working examples" that describe performed experiments, as well as "prophetic examples" that predict various aspects of the invention. By convention, the past tense signals to the patent examiner that the inventors had performed the described work. Inventors must not write a prophetic example in the past tense. An incorrect verb tense transforms the example into a time bomb.

An inappropriate use of the past tense has detonated several biotech patents. A battle over Taq enzyme patent rights revealed that Cetus Corporation inventors had written as a prophetic example in the past tense a detailed enzyme purification protocol. Later, the applicant used that information to distinguish the claimed enzyme from the prior art. Inequitable conduct, a district court judge decided. Another court found that Novo Nordisk engaged in inequitable conduct when its inventors supported the patentability of a claimed biosynthetic growth hormone with "data" from a prophetic example written in the past tense.

Misleading statements may also arise during patent prosecution when inventors file an affidavit called a Rule 132 Declaration to support patentability. While fighting a rejection of patent claims to a glaucoma treatment, a Pharmacia inventor filed a declaration asserting that a certain dose of a prior art compound does not significantly decrease intraocular pressure. The inventor had co-authored an article that contradicted the assertion. The declaration also reported a result from an experiment that had not been performed. The Federal Circuit's finding of inequitable conduct terminated patent rights.

In Aventis v. Amphastar, the Federal Circuit affirmed a finding of inequitable conduct during the prosecution of a patent to a composition of low molecular weight heparins, a drug marketed as Lovenox^® in the United States. The case focused on declarations written by a scientist on behalf of Aventis that distinguished claimed formulations from prior art formulations by their properties. The court found that the scientist had failed to inform the patent examiner that comparisons had been performed with different dosages of the claimed and prior art compounds.

Inequitable Conduct Rulings Incite Judicial Dissent
Judge Rader highlights the traditional view of inequitable conduct in his dissent of the Aventis decision:

"Without doubt, candor and truthful cooperation are essential to an ex parte examination system. With burgeoning application rates, the USPTO must rely on applicant submissions to narrow the prior art search. And, of course, those submissions must be reliable. The threat of inequitable conduct, with its 'atomic bomb' remedy of unenforceability, ensures that candor and truthfulness."

Perhaps, courts are too willing to deploy this bomb. The facts of the Aventis patent, Rader says, support a good faith mistake, not deceptive intent. Rader cautions that the Federal Circuit has relaxed the standards for proclaiming inequitable conduct. His warning echoes Judge Newman's dissent of the McKesson decision: "This court returns to the 'plague' of encouraging unwarranted charges of inequitable conduct, spawning the opportunistic litigation that here succeeded despite consistently contrary precedent."

One day, courts may demand more evidence of deceptive intent before pronouncing inequitable conduct. Until then, inventors should exercise scrupulous vigilance in their dealings with the patent office.

Insect pests remain one of the major constraints to food and fiber production worldwide, despite a range of techniques deployed by farmers to protect their crops. Modern pest control is guided by the principles of integrated pest management (IPM), with pest-resistant germplasm as an important part of the foundation. Biotechnology has allowed the development of novel, genetically modified (GM) crops that express genes from the bacterium Bacillus thuringiensis (Bt) and produce proteins toxic to insects. Since 1996, when the first Bt maize variety was commercialized in the USA, the area planted to insect-resistant Bt varieties has grown dramatically, representing the fastest adoption rate of any agricultural technology in human history. In 2007, insect-resistant Bt corn and cotton plants were grown in 22 countries on 42.1 million hectares (104 million acres).

A newly released book, Integration of Insect-Resistant Genetically Modified Crops within IPM Programs, provides the first comprehensive synthesis of the role of insect-resistant GM crops in crop protection. The book was edited by Jörg Romeis (Agroscope ART, Switzerland), Anthony Shelton (Cornell University, USA), and George Kennedy (North Carolina State University, USA) with the goal of providing an overview of the role insect-resistant GM plants play in different crop systems worldwide. A total of 42 authors from around the globe reviewed the latest available information on insect-resistant GM crops, ranging from their biological and ecological activity to their economic and social impact. The editors hope the book will contribute to a more rational debate about the role GM crops can play in IPM for food and fiber production. Norman Borlaug and Tom Lumpkin, Director General of CYMMT, provide strong endorsements for the book.

ONLINE BIBLIOGRAPHY OF ASSESSMENT STUDIES ON GE CROPS

The International Food Policy Research Institute (IFPRI) has compiled a web-based bibliography of peer-reviewed applied economics literature called bEcon, to assess the impact of genetically engineered (GE) crops in developing economies. bEcon contains 190 articles organized under four major themes that address the different areas of impact: advantages to farmers, consumer preferences and willingness to pay, size and distribution of benefits, and international benefits of trade. The literature is searchable by author, year, and keyword. bEcon is updated every three months, and a CD-ROM is produced on an annual basis for those with limited or no internet access.