How spiders spin silk: Mechanism elegantly explains how spider silk can form so quickly and smoothly

Spider silk is an impressive material; lightweight and stretchy yet stronger than steel. But the challenge that spiders face to produce this substance is even more formidable. Credit: © Tamas Zsebok / Fotolia

Spider silk is an impressive material; lightweight and stretchy yet stronger than steel. But the challenge that spiders face to produce this substance is even more formidable. Silk proteins, called spidroins, must convert from a soluble form to solid fibers at ambient temperatures, with water as a solvent, and at high speed. How do spiders achieve this astounding feat? In new research publishing in the open access journal PLOS Biology on August 5, Anna Rising and Jan Johansson show how the silk formation process is regulated. The work was done at the Swedish University of Agricultural Sciences (SLU) and Karolinska Institutet in collaboration with colleagues in Latvia, China and USA.

Spidroins are big proteins of up to 3,500 amino acids that contain mostly repetitive sequences, but the most important bits for the conversion of spidroins into silk are the ends. 

These terminal regions of the proteins are unique to spider silk and are very similar between different spiders. Spidroins have a helical and unordered structure when stored as soluble proteins in silk glands, but when converted to silk their structure changes completely to one that confers a high degree of mechanical stability. These changes are triggered by an acidity (pH) gradient present between one end of the spider silk gland and the other. The gland proceeds from a narrow tail to a sac to a slender duct, and it is known that silk forms at a precise site within the duct. However, further details of spider silk production have been elusive.

By using highly selective microelectrodes to measure the pH within the glands, the authors showed the pH falls from a neutral pH of 7.6 to an acidic pH of 5.7 between the beginning of the tail and half-way down the duct, and that the pH gradient was much steeper than previously thought. The microelectrodes also showed that the concentration of bicarbonate ions and pressure of carbon dioxide simultaneously rise along the gland. Taken together, these patterns suggested that the pH gradient might form through the action of an enzyme called carbonic anhydrase, which converts carbon dioxide and water to bicarbonate and hydrogen ions (and thereby creating an acidic environment). Using a method developed by the authors, they were able to identify active carbonic anhydrase in the narrower part of the gland and confirm that carbonic anhydrase is indeed responsible for generating the pH gradient.

The authors also found that pH had opposite effects on the stability of the two regions at each end of the spidroin proteins, which was surprising given that these regions had been suggested to have similar roles in silk formation. While one of the ends (the "N-terminal domain") tended to pair up with other molecules at the beginning of the duct and became increasingly stable as the acidity increased along the duct, the other end (the "C-terminal domain") destabilized as the acidity increased, and gradually unfolded until it formed the structure characteristic of silk at the acidic pH of 5.5. These findings show that both ends of the protein undergo dramatic structural changes at the pH found at the beginning of the duct, which is also the point where carbonic anhydrase activity is concentrated.

These insights led the authors to propose a new "lock and trigger" model for spider silk formation, in which gradual pairing up of the N-terminal domains locks spidroins into a network of many protein molecules, while the changes of structure in the C-terminal domains could trigger the rapid polymerization of spidroins into fibers. Interestingly, the structure of the C-terminal domain is similar to those in the "amyloid" fibrils found in the brains of individuals with diseases such as Alzheimer's disease. This mechanism elegantly explains how spider silk can form so quickly and smoothly within the spinning duct of these amazing animals. Besides helping humans to understand how they might mimic the spiders to produce biomimetic spidroin fibers for our own purposes, knowing how spiders spin silk could give insights into natural ways of hindering the amyloid fibrils associated with diseases like dementia.

Source: PLOS

Parasites and the evolution of primate culture

Chimpanzees (stock image). A new study examines the ‘costs’ of innovation, and learning from others. Credit: © shiruikage / Fotolia

Learning from others and innovation have undoubtedly helped advance civilization. But these behaviours can carry costs as well as benefits. And a new study by an international team of evolutionary biologists sheds light on how one particular cost -- increased exposure to parasites -- may affect cultural evolution in non-human primates.

The results, published Dec. 3, 2014 in the journal Proceedings of the Royal Society B, suggest that species with members that learn from others suffer from a wider variety of socially transmitted parasites, while innovative, exploratory species suffer from a wider variety of parasites transmitted through the environment, such as in the soil or water.

"We tend to focus on innovation and learning from others as a good thing, but their costs have received relatively little attention," says McGill University biologist Simon Reader, co-author of the study. "Here, we uncover evidence that socially transmitted pathogen burdens rise with learning from others -- perhaps because close interaction is needed for such learning -- and environmentally transmitted pathogen burdens rise with exploratory behaviour such as innovation and extractive foraging."

Chimpanzees, for example, live in groups and have a wide range of such behaviours, such as digging for food underground or eating new kinds of insects.. Previously, studies have not been able to determine whether costly parasites force primates to engage in more exploratory behaviour -- by diversifying food sources, for example -- or whether exploratory behaviour leads to their having more parasites, Reader notes. "Our results support the idea that exploratory and social behaviours expose primates to specific kinds of parasites."

"The findings also lead to questions about how people and other primates have developed solutions to minimize these parasite costs -- such as eating medicinal plants -- and may help us better understand how the processes underlying human culture arose," Reader says.
The research team, led by Collin McCabe of Harvard University and Charles Nunn of Duke University, based their analyses on databases obtained by surveying thousands of articles on primate behaviour and parasites.

Funding for the research was provided by the National Science Foundation, the Natural Sciences and Engineering Research Council of Canada, and the Netherlands Organisation for Scientific Research.

Source: McGill University

New molecules to burst malaria's bubble

Dr Natalie Spillman. Credit: Alex Maier

Scientists have released details of a raft of new chemicals with potent anti-malarial properties which could open the way to new drugs to fight the disease.

A new paper in PNAS is the third published recently by a group at the Australian National University (ANU). The group has collaborated with scientists from around the globe to uncover potential ammunition in the fight against malaria.

Over 200 million people contract malaria each year, and the parasite that causes the disease has become resistant to most of the drugs currently available.

"The series of papers shows that the malaria parasite has a real Achilles heel, and describe a range of new ways to attack it," said Professor Kiaran Kirk, Dean of the College of Medicine, 
Biology and Environment and one of the scientists involved in the project.

Dr Natalie Spillman, from the Research School of Biology at ANU studied the mechanism by which the parasites are killed.

"The new molecules block a molecular salt pump at the surface of the parasite, causing it to fill up with salt," Dr Spillman said

"This has the effect of drawing water into the parasite, causing it to swell uncontrollably and burst."

Although the process of developing the new compounds into clinical drugs is complex and lengthy, Professor Kirk is optimistic the findings will lead to new treatments.

"It's very early days, but these pump-blocking compounds have some of the most promising anti-malarial potential we've seen," he says.

Aspects of the work were carried out with groups at Griffith University, Monash University and the Menzies School of Health Research in Darwin.

"This is a good example of a long-term, international drug development program in which Australian groups have played a key role," he said.

Source: Australian National University

Using power of computers to harness human genome may provide clues into Ebola virus

Ramaswamy Narayanan, Ph.D., professor in the Charles E. Schmidt College of Science at Florida Atlantic University.

Ramaswamy Narayanan, Ph.D., professor in the Charles E. Schmidt College of Science at Florida Atlantic University, is working to blend the power of computers with biology to use the human genome to remove much of the guesswork involved in discovering cures for diseases.

In an article titled "Ebola-Associated Genes in the Human Genome: Implications for Novel Targets," published in the current MedCrave Online Journal of Proteomics and Bioinformatics, Narayanan describes how key genes that are present in our cells could be used to develop drugs for this disease.

"Bioinformatics is a powerful tool to help us understand biological data," said Narayanan whose research has focused in this field for more than a decade. "We are mining the human genome for Ebola virus association to develop an understanding of the human proteins involved in this disease for subsequent research and development, and to potentially create a pipeline of targets that we can test and evaluate."

Ebola virus disease is a major healthcare challenge facing the globe today and if left unchecked could become a pandemic. A limited knowledgebase exists about the Ebola virus and companies are hastening to develop vaccines and other forms to treat and cure the virus. There are no FDA-approved drugs, and developing vaccines or antibodies and testing them in clinical trials is an arduous process that takes considerable time. Currently, patients infected with Ebola are only able to receive supportive care such as fluid replacement, nutritional support, pain control, and blood pressure maintenance. In some cases, patients may be fortunate enough to be treated with experimental drugs.

Narayanan's work has helped to identify numerous FDA-approved drugs already used for many other diseases including anti-inflammatory drugs, anticoagulants, cancer, HIV, statins and hormones, which could potentially be used to add to the current supportive care for patients with the Ebola virus.

"With the high mortality rate of this disease, the world urgently needs new ways to treat patients," said Narayanan. "The ability to use drugs that are already approved by the FDA could provide clinicians with more options to treat Ebola patients, rather than just relying on supportive measures like fluid replacement or antibiotics."

According to the World Health Organization (WHO), Ebola virus disease (EVD) is a severe, often fatal illness in humans. The virus is transmitted to people from wild animals and spreads in the human population through human-to-human transmissions. The evolving knowledge of this disease is prompting appropriate attention locally and globally. The 2014 Ebola epidemic has affected multiple countries in West Africa with some cases observed in Europe and the United States.

Source: Florida Atlantic University

Nanotechnology against malaria parasites

After maturation, malaria parasites (yellow) are leaving an infected red blood cell and are efficiently blocked by nanomimics (blue). Credit: Fig: modified by University of Basel with permission from ACS

Malaria parasites invade human red blood cells, they then disrupt them and infect others. Researchers at the University of Basel and the Swiss Tropical and Public Health Institute have now developed so-called nanomimics of host cell membranes that trick the parasites. This could lead to novel treatment and vaccination strategies in the fight against malaria and other infectious diseases. Their research results have been published in the scientific journal ACS Nano.

For many infectious diseases no vaccine currently exists. In addition, resistance against currently used drugs is spreading rapidly. To fight these diseases, innovative strategies using new mechanisms of action are needed. The malaria parasite Plasmodium falciparum that is transmitted by the Anopheles mosquito is such an example. Malaria is still responsible for more than 600,000 deaths annually, especially affecting children in Africa (WHO, 2012).
Artificial bubbles with receptors

Malaria parasites normally invade human red blood cells in which they hide and reproduce. They then make the host cell burst and infect new cells. Using nanomimics, this cycle can now be effectively disrupted: The egressing parasites now bind to the nanomimics instead of the red blood cells.

Researchers of groups led by Prof. Wolfgang Meier, Prof. Cornelia Palivan (both at the University of Basel) and Prof. Hans-Peter Beck (Swiss TPH) have successfully designed and tested host cell nanomimics. For this, they developed a simple procedure to produce polymer vesicles -- small artificial bubbles -- with host cell receptors on the surface. The preparation of such polymer vesicles with water-soluble host receptors was done by using a mixture of two different block copolymers. In aqueous solution, the nanomimics spontaneously form by self-assembly.

Blocking parasites efficiently

Usually, the malaria parasites destroy their host cells after 48 hours and then infect new red blood cells. At this stage, they have to bind specific host cell receptors. Nanomimics are now able to bind the egressing parasites, thus blocking the invasion of new cells. The parasites are no longer able to invade host cells, however, they are fully accessible to the immune system.

The researchers examined the interaction of nanomimics with malaria parasites in detail by using fluorescence and electron microscopy. A large number of nanomimics were able to bind to the parasites and the reduction of infection through the nanomimics was 100-fold higher when compared to a soluble form of the host cell receptors. In other words: In order to block all parasites, a 100 times higher concentration of soluble host cell receptors is needed, than when the receptors are presented on the surface of nanomimics.

"Our results could lead to new alternative treatment and vaccines strategies in the future," says Adrian Najer first-author of the study. Since many other pathogens use the same host cell receptor for invasion, the nanomimics might also be used against other infectious diseases. The research project was funded by the Swiss National Science Foundation and the NCCR "Molecular Systems Engineering."

Source: University of Basel

Promising compound rapidly eliminates malaria parasite

A new report says that the rapid action of (+)-SJ733 will likely slow malaria drug resistance. Credit: Peter Barta, St. Jude Children's Research Hospital

An international research collaborative has determined that a promising anti-malarial compound tricks the immune system to rapidly destroy red blood cells infected with the malaria parasite but leave healthy cells unharmed. St. Jude Children's Research Hospital scientists led the study, which appears in the current online early edition of the Proceedings of the National Academy of Sciences (PNAS).

The compound, (+)-SJ733, was developed from a molecule identified in a previous St. Jude-led study that helped to jumpstart worldwide anti-malarial drug development efforts. Malaria is caused by a parasite spread through the bite of an infected mosquito. The disease remains a major health threat to more than half the world's population, particularly children. The World Health Organization estimates that in Africa a child dies of malaria every minute.

In this study, researchers determined that (+)-SJ733 uses a novel mechanism to kill the parasite by recruiting the immune system to eliminate malaria-infected red blood cells. In a mouse model of malaria, a single dose of (+)-SJ733 killed 80 percent of malaria parasites within 24 hours. After 48 hours the parasite was undetectable.

Planning has begun for safety trials of the compound in healthy adults.

Laboratory evidence suggests that the compound's speed and mode of action work together to slow and suppress development of drug-resistant parasites. Drug resistance has long undermined efforts to treat and block malaria transmission.

"Our goal is to develop an affordable, fast-acting combination therapy that cures malaria with a single dose," said corresponding author R. Kiplin Guy, Ph.D., chair of the St. Jude Department of Chemical Biology and Therapeutics. "These results indicate that SJ733 and other compounds that act in a similar fashion are highly attractive additions to the global malaria eradication campaign, which would mean so much for the world's children, who are central to the mission of St. Jude."

Whole genome sequencing of the Plasmodium falciparum, the deadliest of the malaria parasites, revealed that (+)-SJ733 disrupted activity of the ATP4 protein in the parasites. The protein functions as a pump that the parasites depend on to maintain the proper sodium balance by removing excess sodium.

The sequencing effort was led by co-author Joseph DeRisi, Ph.D., a Howard Hughes Medical Institute investigator and chair of the University of California, San Francisco Department of Biochemistry and Biophysics. Investigators used the laboratory technique to determine the makeup of the DNA molecule in different strains of the malaria parasite.

Researchers showed that inhibiting ATP4 triggered a series of changes in malaria-infected red blood cells that marked them for destruction by the immune system. The infected cells changed shape and shrank in size. They also became more rigid and exhibited other alterations typical of aging red blood cells. The immune system responded using the same mechanism the body relies on to rid itself of aging red blood cells.

Another promising class of antimalarial compounds triggered the same changes in red blood cells infected with the malaria parasite, researchers reported. The drugs, called spiroindolones, also target the ATP4 protein. The drugs include NITD246, which is already in clinical trials for treatment of malaria. Those trials involve investigators at other institutions.

"The data suggest that compounds targeting ATP4 induce physical changes in the infected red blood cells that allow the immune system or erythrocyte quality control mechanisms to recognize and rapidly eliminate infected cells," DeRisi said. "This rapid clearance response depends on the presence of both the parasite and the investigational drug. That is important because it leaves uninfected red blood cells, also known as erythrocytes, unharmed."

Laboratory evidence also suggests that the mechanism will slow and suppress development of drug-resistant strains of the parasite, researchers said.

Planning has begun to move (+)-SJ733 from the laboratory into the clinic beginning with a safety study of the drug in healthy adults. The drug development effort is being led by a consortium that includes scientists at St. Jude, the Swiss-based non-profit Medicines for Malaria Venture and Eisai Co., a Japanese pharmaceutical company.

Source: St. Jude Children's Research Hospital

Potential biological control for avocado-ravaging disease

University of Florida scientists think they’ve found the first potential biological control strategy against laurel wilt, a disease that threatens Florida’s avocado industry. The redbay ambrosia beetle, see here, bores holes into avocado trees, bringing the disease that causes laurel wilt. Credit: Lyle Buss, UF/IFAS

University of Florida scientists believe they've found what could be the first biological control strategy against laurel wilt, a disease that threatens the state's $54 million-a-year avocado industry.

Red ambrosia beetles bore holes into healthy avocado trees, bringing with them the pathogen that causes laurel wilt. Growers control the beetles that carry and spread laurel wilt by spraying insecticides on the trees, said Daniel Carrillo, an entomology research assistant professor at the Tropical Research and Education Center in Homestead.

But a team of researchers from the Tropical REC and the Indian River Research and Education Center in Fort Pierce have identified a potential biological control to use against redbay ambrosia beetles that could help growers use less insecticide.

First, they exposed beetles to three commercially available fungi, and all of the beetles died. Then they sprayed the fungi on avocado tree trunks, and beetles got infected while boring into the trunk. About 75 percent of those beetles died, said Carrillo, an Institute of Food and Agricultural Sciences faculty member.

Ideally, the fungal treatments could prevent beetles from boring into the trees, eliminating the risk that the pathogen would enter the trees, the study said. But tests showed female beetles bored into the trees and built tunnels regardless of the treatment. Still, researchers say their treatment can prevent the female beetles from laying eggs.

UF/IFAS scientists don't know yet how much less chemical spray will be needed to control the redbay ambrosia beetle. But Carrillo sees this study as the first step toward controlling the beetle in a sustainable way.

"When you want to manage a pest, you want an integrated pest management approach," Carrillo said. "This provides an alternative that we would use in combination with chemical control."

The redbay ambrosia beetle -- native to India, Japan, Myanmar and Taiwan -- was first detected in 2002 in southeast Georgia. It was presumably introduced in wood crates and pallets, and its rapid spread has killed 6,000 avocado trees in Florida, or about 1 percent of the 655,000 commercial trees in Florida. The beetle was first discovered in South Florida in 2010.

Most American-grown avocados come from California, with the rest coming from Florida and Hawaii. The domestic avocado market is worth $429 million, according to Edward Evans, a UF associate professor of food and resource economics, also at the Tropical REC. Florida's avocados are valued at about $23 million, or about 5 percent of the national market.

The redbay ambrosia beetle is not an issue with California avocados, so the new tactic found by Florida scientists wouldn't apply to this pest in the Golden State, said Mark Hoddle, a biological control Extension specialist with the University of California-Riverside. Hoddle studies biological pest control for California avocados. Scientists there are exploring ways to control a different ambrosia beetle, he said, and bug-killing fungi may be useful for the new California pest.

More than 95 percent of Florida's commercial avocados grow in Miami-Dade County, although many Floridians have avocado trees in their yard.
The redbay ambrosia beetle feeds and reproduces on a very wide variety of host plants, native oaks, sycamores, and of course it is very detrimental to avocados.

Source:University of Florida Institute of Food and Agricultural Sciences

Scientists find 240-million-year-old parasite that infected mammals' ancestor

Scott Gardner. Credit: Craig Chandler, University Communications.

An egg much smaller than a common grain of sand and found in a tiny piece of fossilized dung has helped scientists identify a pinworm that lived 240 million years ago.

It is believed to be the most ancient pinworm yet found in the fossil record.

The discovery confirms that herbivorous cynodonts -- the ancestors of mammals -- were infected with the parasitic nematodes. It also makes it even more likely that herbivorous dinosaurs carried pinworms.

Scott Gardner, a parasitologist and director of the Harold W. Manter Laboratory of Parasitology at the University of Nebraska-Lincoln, was among an international group of scientists who published the study in the journal Parasites & Vectors.

"This discovery represents a first for our team and I think it opens the door to finding additional parasites in other species of fossil organisms," he said.

The team found the pinworm egg in a coprolite -- fossilized feces -- collected in 2007 at an excavation site in Rio Grande do Sul state in southern Brazil.

The coprolite was collected at a site with abundant fossilized remains of cynodonts. Previously, an Ascarid-like egg -- resembling a species of nematode commonly found in modern-day mammals -- was found in the coprolite.

The pinworm egg, representing an undescribed or "new species," was named Paleoxyuriscockburni, in honor of Aidan Cockburn, founder of the Paleopathology Association.

The structure of the pinworm egg placed it in a biological group of parasites that occur in animals that ingest large amounts of plant material. Its presence helped scientists deduce which cynodont species, of several found at the collection site, most likely deposited the coprolite.

Since the field of paleoparasitology, or the study of ancient parasites, emerged in the early 20th century, scientists have identified parasites of both plants and animals that date back as far as 500 million years ago.

The study of parasites in ancient animals can help determine the age of fossilized organisms and help establish dates of origin and diversification for association between host species and parasites. Coprolites are a key part of the study, enabling a better understanding of the ecological relationships between hosts and parasites.

Other members of the team were Jean-Pierre Hugot of the National Museum of Natural History in Paris; Victor Borba, Juliana Dutra, Luiz Fernando Ferreira and Adauto Araujo of Oswaldo Cruz Foundation in Rio de Janeiro; Prisiclla Araujo and Daniela Leles of Fluminense Federal University in Rio de Janeiro; and Atila August Stock Da-Rosa of the Federal University of Santa Maria in Rio Grande do Sul.

Source: University of Nebraska-Lincoln