Stopping a wound from bleeding is essential for human health. Blood coagulation – in which blood goes from liquid to gel and forms a clot – can prevent excessive bleeding and infection. But exactly what molecular events transpire when blood coagulates has remained somewhat mysterious. Using a new profiling procedure invented by investigators from Brigham and Women’s Hospital permitted them to elucidate the role of immunoresolvents – molecules that help resolve inflammation and infections –in blood coagulation, identifying a new cluster of these molecules that are produced when blood coagulates.
“We’ve identified factors biosynthesized by human blood coagulation that elicit immune responses that protect the host,” said corresponding author Charles N. Serhan, PhD, DSc. director and principal investigator at the Center for Experimental Therapeutics and Reperfusion Injury at BWH. “Our results uncover a previously uncharacterized connection between the coagulation of blood and innate host defence mechanisms. We’ve demonstrated for the first time how the innate immune response is connected to coagulation via novel pro-resolving mediators.”
The new profiling procedure allowed the team to identify a cluster of immunoresolvents, namely resolvin D1, resolvin D5, resolvin E1, lipoxin B4 and maresin 1. These molecules activate immune cells called phagocytes, which can engulf and kill bacteria in the blood. Treating human blood with the components of this cluster of molecules discovered at BWH enhanced the abilities of phagocytes and helped the immune system attack E. coli, a common source of bacterial infection.
Interestingly, the newly developed profiling technique holds potential for profiling immunoresolvants in many contexts. The current study offers a glimpse of immunoresolvents found in the blood of healthy individuals, but the researchers are also interested in studying blood samples from patients with sepsis to pinpoint differences in immunoresolvents. Beyond blood, the research team also found a distinct profile of immunoresolvents in samples of healthy versus cancerous tissue from the testes – they note that this new profiling technique could potentially be used in the future to help distinguish between cancerous and healthy tissue from the testes or elsewhere in the body.

Brigham and Women’s Hospital
bwhclinicalandresearchnews.org/2017/08/04/whats-new-in-research-august-2017-2/
 

Neuroscientists have uncovered the genetic basis for why many long-term survivors of childhood cancer develop meningiomas, the most common adult brain tumour, decades after their treatment with cranial radiation.
The findings show that radiation causes genetic rearrangements in DNA that result in meningiomas, say co-principal investigators Gelareh Zadeh, neurosurgeon-scientist, Head of Surgical Oncology, and Ken Aldape, neuropathologist-scientist, Director, MacFeeters-Hamilton Neuro-Oncology Research Program, Princess Margaret Cancer Centre, University Health Network.
Dr. Zadeh is an Associate Professor, Division of Neurosurgery, and holds the Wilkins Family Chair in Brain Tumor Research; and Dr. Aldape, Professor, Laboratory Medicine and Pathobiology, both at University of Toronto.
The study compared and contrasted the biology of radiation-induced meningiomas (RIMs) to those that appear sporadically in the general population.
"Radiation-induced meningiomas appear the same on MRI and pathology, feel the same during surgery and look the same under the operating microscope. What’s different is they are more aggressive, tend to recur in multiples and invade the brain, causing significant morbidity and limitations (or impairments) for individuals who survive following childhood radiation," says Dr. Zadeh.
The research team analysed RIMs from patients who had received cranial-spinal radiation as children; the majority of whom (74 per cent) had survived either leukaemia or paediatric brain cancer. The study also showed that RIMs developed regardless of the radiation dose by collaborating with scientists in Germany where low-dose radiation was a common treatment many years ago for scalp ringworm.
"By understanding the biology, the goal is to identify a therapeutic strategy that could be implemented early on after childhood radiation to prevent the formation of these tumours in the first place," says Dr. Zadeh.
Dr. Aldape says: "It is an important clinical problem because it presents a paradoxical dilemma that while cranial-spinal radiation is needed to cure many childhood cancers, an unfortunate consequence is that 10-to-15-years following radiation treatment some survivors develop meningiomas.
"Our research identified a specific rearrangement involving the NF2 gene that causes radiation-induced meningiomas. But there are likely other genetic rearrangements that are occurring as a result of that radiation-induced DNA damage. So one of the next steps is to identify what the radiation is doing to the DNA of the meninges."
He adds: "In addition, identifying the subset of childhood cancer patients who are at highest risk to develop meningioma is critical so that they could be followed closely for early detection and management."

Princess Margaret Cancer Centre, University Health Network
www.uhn.ca/corporate/News/PressReleases/Pages/brain-tumour-scientists-map-mutation-that-drives-tumours-in-childhood-cancer-survivors-treated-with-radiation-decades-earl.aspx

Researchers have found a specific type of immune regulatory cells that could soon be used as potential clinical biomarkers to diagnose certain autoimmune diseases.
The team from Instituto de Medicina Molecular (iMM) Lisboa, led by Luis Graça, analysed blood samples from Sjögren syndrome patients, an autoimmune disease that affects the mucous membranes and moisture-secreting glands of the eyes and mouth, and found that these patients have a significant increase in a specific type of immune cells called T follicular regulatory cells (Tfr). 
These cells are usually found in lymphoid tissues where they regulate antibody production. It was a surprise to find an increase of these type of cells in patients with excessive antibody production. In fact, the results were the opposite of what the team was expecting.
To understand the reason behind such unexpected results the researchers studied different biological samples. For instance, comparing Tfr cells in the blood and in the tissues where antibodies are produced (tonsils obtained from children subjected to tonsillectomies), provided evidence that blood Tfr cells are immature, not able to fully suppress antibody production. Such immaturity was confirmed by studying blood samples from other patients with genetic defects. Furthermore, exposure of healthy volunteers to flu vaccine led to an increase in blood Tfr cells, in line with their generation during immune responses with antibody production.
Blood circulating Tfr cells are distinguished from other circulating lymphocytes by two molecular markers, CXCR5 and FOXP3, the first of which endows these cells with the ability to migrate into specific zones of lymph nodes where they may complete maturation and regulate antibody production.
The team is now trying to understand what happens to these cells in other autoimmune diseases to evaluate their potential not only for diagnostic but also to identify which patients may benefit with medicines that interfere with the production of harmful antibodies.

Instituto de Medicina Molecular (iMM) Lisboa
imm.medicina.ulisboa.pt/en/imm-lisboa/news/archive/novo-tipo-de-celulas-do-sangue-funcionam-como-indicadores-de-doencas-autoimunes/

Every year, just over 1000 people are diagnosed with kidney cancer in Sweden. The three most common variants are clear cell, papillary and chromophobe renal cancer. Researchers compare the gene expression in tumour cells from a kidney cancer patient with cells from healthy tissue to figure out in which part of the kidney the cancer began and what went wrong in these cells. Now, a research team at Lund University in Sweden has discovered that in the Cancer Genome Atlas database, the gene expression in reference samples from normal tissue varies, depending on where in the kidney the samples happen to have been taken. The analyses can be improved by clarifying which samples correspond to the correct tissue.
The part of the kidney which purifies the blood and generates urine is called the nephron and functions as a kind of tubing system. Each kidney contains around a million nephrons which collectively filter 180 litres of primary urine (waste products, water and salts) every day. This results in 1.5 litres of concentrated liquid, which is excreted through urination.
“Everything is very specifically regulated and the cells have different gene expression and hence properties depending on their location in the tubing.”, explains Håkan Axelson, research team leader and professor of molecular tumour biology.
When a tumour biopsy is taken from a patient and compared with healthy kidney tissue, it serves to map how the various genes are expressed so as to clarify what has gone wrong in the tumour cells.  The Cancer Genome Atlas – an international database containing almost 1000 samples from kidney tumours and healthy tissue – is a tool in this process.
“But when our research team studied the samples from the database, we noticed a great range of gene expressions between normal tissue samples. It emerged that the samples in the Cancer Genome Atlas were taken at different depths in the kidney and therefore contain different types of cells, which means that their gene expressions also vary”, says Håkan Axelson.
The normal reference samples thus contain various types of cells depending on where in the kidney they happen to have been taken. Since the Atlas does not state the location in the kidney the reference sample was collected, the comparison risks being unreliable and sometimes completely incorrect.
“Since the gene expression in the cells varies depending on their location, it is important that the normal samples contained in the database should also be taken from the location corresponding to that of the patient’s tumour”, says David Lindgren, who is the lead author of the study.
As an example, it was previously suspected that clear cell tumours occur in the first part of the nephron, but if these tumour cells are compared with a normal sample taken deeper inside the nephron, the cells will not correspond to the tumour sample. The gene expression is thereby different. Although each patient is unique, the various types of tumours have different specific genetic changes which occur as a consequence of properties in the cell in which the tumour originated.
“It is extremely important to know what characterises the cells in which the tumour occurs. Through better understanding of this interaction, we can increase our understanding of the course of the disease, which could be significant for diagnostics and, in the longer term, also for the choice of treatment”, concludes Håkan Axelson.

Lund University
www.lunduniversity.lu.se/article/improved-analysis-of-kidney-cancer

Currently, testing for Zika requires that a blood sample be refrigerated and shipped to a medical centre or laboratory, delaying diagnosis and possible treatment. Although the new proof-of-concept technology has yet to be produced for use in medical situations, the test’s results can be determined in minutes. Further, the materials required for the test do not require refrigeration and may be applicable in testing for other emerging infectious diseases.
The researchers tested blood samples taken from four people who had been infected with Zika virus and compared it to blood from five people known not to have the virus. Blood from Zika-infected patients tested positive, but blood from Zika-negative controls did not. The assay produced no false-positive results.
Among the reasons such a test is needed, according to the researchers, is that many people infected with Zika don’t know they’re infected. Although symptoms include fever, joint pain, muscle pain and rash, many people don’t feel ill after being bitten by an infected mosquito. Testing is particularly important for pregnant women because Zika infection can cause congenital Zika syndrome, which contributes to several neurologic problems in the foetus or newborn infant.
“Zika infection is often either asymptomatic or mildly symptomatic,” said Evan D. Kharasch, MD, PhD, one of the study’s three senior investigators. “The most effective way to diagnose the disease is not to wait for people to develop symptoms but to do population screening.”
That strategy requires inexpensive, easy-to-use and easy-to-transport tests. Kharasch, the Russell D. and Mary B. Shelden Professor of Anesthesiology, collaborated with Srikanth Singamaneni, PhD, an associate professor of mechanical engineering & materials science, and Jeremiah J. Morrissey, PhD, a research professor of anesthesiology, to create the test, which uses gold nanorods mounted on paper to detect Zika infection within a few minutes.
“If an assay requires electricity and refrigeration, it defeats the purpose of developing something to use in a resource-limited setting, especially in tropical areas of the world,” said Singamaneni. “We wanted to make the test immune from variations in temperature and humidity.”
The test relies on a protein made by Zika virus that causes an immune response in infected individuals. The protein is attached to tiny gold nanorods mounted on a piece of paper. The paper then is completely covered with tiny, protective nanocrystals. The nanocrystals allow the diagnostic nanorods to be shipped and stored without refrigeration prior to use.
To use the test, a technician rinses the paper with slightly acidic water, removing the protective crystals and exposing the protein mounted on the nanorods. Then, a drop of the patient’s blood is applied. If the patient has come into contact with the virus, the blood will contain immunoglobulins that react with the protein.
“We’re taking advantage of the fact that patients mount an immune attack against this viral protein,” said Morrissey. “The immunoglobulins persist in the blood for a few months, and when they come into contact with the gold nanorods, the nanorods undergo a slight color change that can be detected with a hand-held spectrophotometer.
“With this test, results will be clear before the patient leaves the clinic, allowing immediate counselling and access to treatment.”
The colour change cannot be seen with the naked eye, but the scientists are working to change that. They’re also working on developing ways to use saliva rather than blood.
Although the test uses gold, the nanorods are very small. The researchers estimate that the cost of the gold used in one of the assays would be 10 to 15 cents.
As other infectious diseases emerge around the world, similar strategies potentially could be used to develop tests to detect the presence of viruses that may become problematic, according to the researchers.

Washington University School of Medicine
medicine.wustl.edu/news/test-uses-nanotechnology-quickly-diagnose-zika-virus/