New possibilities for hydrogen-producing algae

Photosynthesis produces the food that we eat and the oxygen that we breathe ― could it also help satisfy our future energy needs by producing clean-burning hydrogen? Researchers studying a hydrogen-producing, single-celled green alga,Chlamydomonas reinhardtii, have unmasked a previously unknown fermentation pathway that may open up possibilities for increasing hydrogen production.

C. reinhartii, a common inhabitant of soils, naturally produces small quantities of hydrogen when deprived of oxygen. Like yeast and other microbes, under anaerobic conditions this alga generates its energy from fermentation. During fermentation, hydrogen is released though the action of an enzyme called hydrogenase, powered by electrons generated by either the breakdown of organic compounds or the splitting of water by photosynthesis. Normally, only a small fraction of the electrons go into generating hydrogen. However, a major research goal has been to develop ways to increase this fraction, which would raise the potential yield of hydrogen.

In the new study by Dubini et al*, published in the Journal of Biological Chemistry, researchers at the Carnegie Institution's Department of Plant Biology, the National Renewable Energy Laboratory (NREL), and the Colorado School of Mines (CSM), examined metabolic processes in a mutant strain that was unable to assemble an active hydrogenase enzyme. The researchers, who include Alexandra Dubini (NREL), Florence Mus (Carnegie), Michael Seibert (NREL), Matthew Posewitz (CSM), and Arthur Grossman (Carnegie), expected the cell's metabolism to compensate by increasing metabolite flow along other known fermentation pathways, such as those producing formate and ethanol as end products. Instead, the algae activated a pathway leading to the production of succinate, which was previously not associated with fermentation metabolism in C. reinhardtii. Notably, succinate, a widely used industrial chemical normally synthesized from petroleum, is included in the Department of Energy's list of the top 12 value added chemicals from biomass.

"We actually didn't know that this particular pathway for fermentation metabolism existed in the alga until we generated the mutant," says Carnegie's Arthur Grossman. "This finding suggests that there is significant flexibility in the ways that soil-dwelling green algae can metabolize carbon under anaerobic conditions. By blocking and modifying some of these metabolic pathways, we may be able to augment the donation of electrons to hydrogenase under anaerobic conditions and produce elevated levels of hydrogen."

Grossman points out that it makes evolutionary sense that a soil organism such as Chlamydomonas would have a variety of metabolic pathways at its disposal. Oxygen levels, nutrient availability, and levels of metals and toxins can be extremely variable in soils, over both the short and long term. "In such an environment", Grossman says, "these organisms must evolve flexible metabolic circuits; the variety of conditions to which the organisms are exposed might favor one pathway for energy metabolism over another, which would help the organism compete in the soil environment over evolutionary time."

Grossman led the effort to generate a fully sequenced Chlamydomonas genome, which has allowed researchers to identify key genes encoding proteins involved in both fermentation and hydrogen production. Grossman feels that it is of immediate importance to generate new mutant strains to help us understand how we may be able to alter fermentation metabolism and the production of hydrogen. NREL's Michael Seibert, the project's Principal Investigator, observed that "the overarching goal of the work is to gain a fundamental understanding of the total suite of metabolic processes occurring in Chlamydomonas and how they interact; this discovery effort will lead to the development of novel ways to produce renewable hydrogen and other biofuels, which will benefit all of us".

"These are really exciting times in the field," says Matthew Posewitz. "The tools developed at Carnegie and by other groups in the field are presenting unprecedented opportunities for scientists to make important advances in our understanding of the basic biology of organisms such as Chlamydomonas."

As an energy source to potentially replace fossil fuels, hydrogen would greatly reduce the emission of greenhouse gases. Proponents of algal-based hydrogen production point out that, unlike ethanol produced from crops, it would not compete with food production for agricultural land.

Implants mimic infection to rally immune system against tumors

Bioengineers at Harvard University have shown that small plastic disks impregnated with tumor-specific antigens and implanted under the skin can reprogram the mammalian immune system to attack tumors.

The research -- which ridded 90 percent of mice of an aggressive form of melanoma that would usually kill the rodents within 25 days -- represents the most effective demonstration to date of a cancer vaccine.

Harvard's David J. Mooney and colleagues describe the research in the current issue of the journal Nature Materials.

"Our immune systems work by recognizing and attacking foreign invaders, allowing most cancer cells -- which originate inside the body -- to escape detection," says Mooney, Gordon McKay Professor of Bioengineering in Harvard's School of Engineering and Applied Sciences. "This technique, which redirects the immune system from inside the body, appears to be easier and more effective than other approaches to cancer vaccination."

Most previous work on cancer vaccines has focused on removing immune cells from the body and reprogramming them to attack malignant tissues. The altered cells are then reinjected back into the body. While Mooney says ample theoretical work suggests this approach should work, in experiments more than 90 percent of the reinjected cells have died before having any effect.

The implants developed by Mooney and colleagues are slender disks measuring 8.5 millimeters across. Made of an FDA-approved biodegradable polymer, they can be inserted subcutaneously, much like the implantable contraceptives that can be placed in a woman's arm.

The disks are 90 percent air, making them highly permeable to immune cells. They release cytokines, powerful attractants of immune-system messengers called dendritic cells.

These cells enter an implant's pores, where they are exposed to antigens specific to the type of tumor being targeted. The dendritic cells then report to nearby lymph nodes, where they activate the immune system's T cells to hunt down and kill tumor cells throughout the body.

"Much as an immune response to a bacterium or virus generates long-term resistance to that particular strain, we anticipate our materials will generate permanent and body-wide resistance against cancerous cells, providing durable protection against relapse," says Mooney, a core member of the recently established Wyss Institute for Biologically Inspired Engineering at Harvard.

The implants could also be loaded with bacterial or viral antigens to safeguard against an array of infectious diseases. They could even redirect the immune system to combat autoimmune diseases such as type 1 diabetes, which occurs when immune cells attack insulin-producing pancreatic cells.

"This study demonstrated a powerful new application for polymeric biomaterials that may potentially be used to treat a variety of diseases by programming or reprogramming host cells," Mooney and his co-authors write in Nature Materials. "The system may be applicable to other situations in which it is desirable to promote a destructive immune response (for example, eradicate infectious diseases) or to promote tolerance (for example, subvert autoimmune disease)."

Topical treatment wipes out herpes with RNAi

Whether condoms or abstinence, most efforts to prevent sexually transmitted diseases have a common logic: keep the pathogen out of your body altogether. While this approach is certainly reasonable enough, it doesn't help the countless people worldwide who, for a number of reasons, are not in a position to control their sexual circumstances.

Now, Harvard Medical School professor of pediatrics Judy Lieberman, who is also a senior investigator at the Immune Disease Institute, has overseen the development of a topical treatment that, in mice, disables key genes necessary for herpesvirus transmission. Using a laboratory method called RNA interference, or RNAi, the treatment cripples the virus in a molecular two-punch knockout, simultaneously disabling its ability to replicate, as well as the host cell's ability to take up the virus.

What's more, the treatment is just as effective when applied anywhere from one week prior to a few hours after exposure to the virus. In that sense, the basic biology of this prophylactic enables a real-world utility.

"People have been trying to make a topical agent that can prevent transmission, a microbicide, for many years," says Lieberman. "But one of the main obstacles for this is compliance. One of the attractive features of the compound we developed is that it creates in the tissue a state that's resistant to infection, even if applied up to a week before sexual exposure. This aspect has a real practicality to it. If we can reproduce these results in people, this could have a powerful impact on preventing transmission."

These findings will be published in the January 22 issue of Cell Host and Microbe.

The World Health Organization estimates that approximately 536 million people worldwide are infected with herpes simplex virus type 2 (HSV-2), the most common strain of this sexually transmitted disease. Women are disproportionately affected. This is especially serious, since the virus can easily be passed from mother to child during birth, and untreated infants face risks of brain damage and death. While HSV-2 alone isn't life-threatening in adults, infection does increase a person's vulnerability to other viruses such as HIV.

In order for the herpesvirus to infect the host, two conditions must be met. First, the virus must be able to enter and take over host cells. Second, the virus must then reproduce itself. Lieberman's topical treatment uses RNAi to foil both these events.

RNAi, a biological process that was identified barely a decade ago, has transformed the field of biological research. A breakthrough that earned the Nobel Prize in 2006, RNAi is a natural cellular process that occurs in all cells of all multicellular organisms to regulate the translation of genetic information into proteins. This natural process can be manipulated by researchers to switch off specific genes, and there is much research and development work to harness RNAi for therapeutics.

Many in the field think RNAi-based drugs may be the next important new class of drugs.

By introducing tiny RNA molecules into cells, researchers can target a gene of interest and, in effect, throw a wrench into that gene's ability to build protein molecules. For all intents and purposes, that gene is now disabled.

While RNAi has profoundly accelerated the ability of scientists to probe and interrogate cells in the Petri dish, therapeutic breakthroughs have proved far more problematic. Researchers have had a difficult time delivering these tiny RNA molecules and ensuring that they actually penetrate the desired cells and tissues in a living organism.

Modifying a delivery technique that Lieberman developed in 2005, she and postdoctoral fellow Yichao Wu and junior researcher Deborah Palliser (who now heads her own laboratory at Albert Einstein College of Medicine) treated mice with strands of RNA that were fused to cholesterol molecules, which made it possible for the molecules to pass through the cell membranes. When applied in the form of a topical solution, these RNA molecules could then be fully absorbed into the vaginal tissue, protecting the mice against a lethal dose of administered virus.

One RNA molecule in the topical solution targeted a herpes gene called UL29, which the virus needs to replicate. Knocking out UL29 inactivates the virus.

Another RNA molecule targeted Nectin-1, a surface protein found on cells in the vaginal tissue. Nectin-1 acts as a kind of host gatekeeper to which the virus binds to pass into the cell. Without Nectin-1, the virus simply can't infect cells.

Either RNA molecule delivered by itself would be sufficient to block the virus, but together in this RNAi cocktail, the host tissue becomes like a fortress that pulls up the drawbridge to block the enemy's entrance, and also has a full-fledged battle plan to slaughter the enemy if they make it through.

"As far as we could tell, the treatment caused no adverse effects, such as inflammation or any kind of autoimmune response," says Lieberman. "And while knocking out a host gene can certainly be risky, we didn't see any indication that temporarily disabling Nectin-1 interfered with normal cellular function."

New sperm shaker to improve IVF success

Scientists have developed a ground-breaking method for testing the quality of a sperm before it is used in IVF and increase the chances of conception.

Researchers at the University of Edinburgh, funded by the EPSRC (Engineering and Physical Sciences Research Council), have created a way of chemically 'fingerprinting' individual sperm to give an indication of quality. Scientists can then consider whether the sperm is healthy enough to be used to fertilise an egg as part of an IVF treatment.

The sperm are captured in two highly focussed beams of laser light. Trapped in what are essentially 'optical tweezers', an individual sperm's DNA properties are identified by the pattern of the vibrations they emit in a process known as Raman spectroscopy. This is the first time this process has been used to evaluate DNA damage in sperm.

Dr Alistair Elfick, lead scientist on the project, said: "In natural conception the fittest and healthiest sperm are positively selected by the arduous journey they make to the egg. What our technology does is to replace natural selection with a DNA based 'quality score'. But this is not about designer babies. We can only tell if the sperm is strong and healthy not if it will produce a baby with blue eyes."

In the past quality tests of sperm have mostly been carried out on the basis of shape and activity. While these do give some indication of health of the sperm they do not give its DNA status.

There are established tests for sperm DNA quality but they work by cutting the cells in half and tagging them with fluorescent dye – a process that kills the sperm and renders it useless. This new process does not destroy the sperm, so if it is found to have good DNA quality, it can still be used in IVF treatment.

Conception rates in both IVF treatment and intercourse are at around one in four. By selecting the best quality sperm it is hoped this new process could both increase a couple's chances of conception and give the child the best potential start in life.

The research is currently in a pre-clinical phase, and if successful could be available to patients in the next five to ten years.