Post-conception mutations may play an important role in autism

Over the past decade, mutations in more than 60 different genes have been linked with autism spectrum disorder, including de novo mutations, which occur spontaneously and aren’t inherited. But much of autism still remains unexplained.
A new study of nearly 6,000 families implicates a hard-to-find category of de novo mutations: those that occur after conception and therefore affect only a subset of cells.
De novo mutations can occur in a parent’s sperm or egg. Alternately, they can occur after egg and sperm meet, arising in an embryonic cell. These are known as somatic mutations or post-zygotic mutations (PZMs).
If a PZM happens very early, when the embryo has just a handful of cells, the mutation will show up in most of the mature organism’s cells. But the later PZMs occur during embryonic development, the fewer cells will carry them, making them harder to detect.
“If the mutation is in a very small fraction of all cells, it will be missed by whole-exome sequencing,” said Elaine Lim, a postdoctoral fellow in the lab of Christopher A. Walsh, the Bullard Professor of Pediatrics and Neurology at Harvard Medical School and Boston Children’s Hospital. Lim is first author of the study; Walsh is the senior investigator.
To identify PZMs, Lim, Walsh and colleagues obtained whole-exome sequencing data previously gathered from 5,947 families, usually through blood tests, courtesy of the Simons Foundation Autism Research Initiative Simplex Collection, the Autism Sequencing Consortium and Autism Speaks. They then re-sequenced some of the DNA from these children using three independent sequencing technologies in parallel.
Based on their findings, they classified 7.5 percent of autism spectrum disorder subjects’ de novo mutations as PZMs. Of these, 83 percent had not been picked up in the original analysis of their genome sequence.
Some PZMs affected genes already known to be linked to autism or other neurodevelopmental disorders (such as SCN2A, HNRNPU and SMARCA4) but sometimes affected these genes in different ways. Many other PZMs occurred in genes known to be active in brain development (such as KLF16 and MSANTD2) but not previously associated with autism spectrum disorder.
The connection of these genes to autism may have been missed because the earlier studies focused on mutations that knocked down gene function, the authors said.
“Some of the postzygotic mutations we found represented a gain of function, not a loss of function,” said Lim, who is also affiliated with the Wyss Institute for Biologically Inspired Engineering.
Lim, Walsh and colleagues then brought in another huge data set: gene expression data from the BrainSpan project. These publicly available data came from autopsies of brain samples from deceased patients of different ages, from prenatal through adult.
Comparing these with the genomic sequencing data, based mostly on blood DNA samples, allowed the researchers to estimate the timing of the PZMs and the brain regions they affected.
“By overlapping the data, we can start to map where in the brain these genes are expressed and when the mutations occurred during development,” said Lim. These analyses showed that PZMs in the subjects with autism spectrum disorder occur disproportionately in genes expressed in the amygdala.
“This was exciting to us, in that the amygdala has been proposed as an important region of the brain in autism,” said Lim.
Overall, the work adds to the evidence that complex brain disorders, such as epilepsy, intellectual disability, schizophrenia and brain malformations, can arise from non-inherited mutations that occur at some point during prenatal development.
Harvard Medical Schoolhttp://tinyurl.com/yb2lpxd7

Biosensor could help diagnose illnesses directly in serum

In this age of fast fashion and fast food, people want things immediately. The same holds true when they get sick and want to know what’s wrong. But performing rapid, accurate diagnostics on a serum sample without complex and time-consuming manipulations is a tall order. Now, a team reports that they have developed a biosensor that overcomes these issues.
Field-effect transistor (FET)-based biosensors are ideal for point-of-care diagnostics because they are inexpensive, portable, sensitive and selective. They also provide results quickly and can be mass-produced to meet market demand. These sensors detect the change in an electric field that results from a target compound, such as a protein or DNA, binding to it. But serum has a high ionic strength, or a high concentration of charged ions, that can mask the targets. Previous research has reported use of pre-treatment steps, complex devices, and receptors with different lengths and orientations on the sensor surface, but with limited success. Alexey Tarasov and colleagues wanted to develop a new approach that would make it easier for FETs to be made as point-of-care diagnostic devices for serum analyses.
The researchers developed a FET sensor that included antibody fragments and polyethylene glycol molecules on a gold surface, which they linked to a commercially available transducer. In this configuration, different sensor chips can be swapped out for use with the same transducer. As a proof-of-principle, they tested the sensor with human thyroid-stimulating hormone. The team found that they could detect the hormone at sub-picomolar concentrations, well below the detection limit previously reported with FETs, when testing it at elevated temperatures. They say that the device could be modified to diagnose many conditions and illnesses, and is inexpensive and easy to use.
American Chemical Society http://tinyurl.com/y99pwak6

Researchers validate a clinical test for fusion genes

An assay that identifies a peculiar but important abnormality in cancer cells has been developed and validated by researchers at The Ohio State University Comprehensive Cancer Center – Arthur G. James Cancer Hospital and Richard J. Solove Research Institute (OSUCCC – James).
The assay, called OSU-SpARKFuse (Ohio State University-Spanning Actionable RNA Kinase Fusions), detects a genetic change called gene fusions in solid tumours.
Gene fusions happen when parts of two different genes join together. Gene fusions can happen, for example, when a piece of one chromosome becomes attached to another. Such chromosome “translocations” can join two genes that together become a major driver of cancer-cell and tumour growth.
Targeted therapies are becoming increasingly available that block the activity of fusion genes, particular those involving kinase genes. Whereas current assays for detecting gene fusions require previous knowledge of both genes involved in the fusion, OSU-SpARKFuse was designed to accurately detect fusions when only one of the genes is known, which allows for the discovery of novel gene fusions.
“We designed OSU-SpARKFuse to meet these needs and to identify patients who are eligible for novel therapies such as FGFR inhibitors or NTRK inhibitors that target gene fusions,” says principal investigator Sameek Roychowdhury, MD, PhD, assistant professor in the Division of Medical Oncology at Ohio State.
“Along with detecting gene fusions, OSU-SpARKFuse can provide gene-expression analysis, detect single-nucleotide changes and identify alternative splicing events and resistance genes,” says first author Julie Reeser, PhD, technical supervisor of the OSUCCC – James Cancer Genomics Laboratory.
“Additionally, OSU-SpARKFuse does not require information regarding the location of the fusion in each gene. It is an accurate, reproducible, cost-effective assay that detects gene fusions across many genes and from the small samples of tumour tissue obtained by biopsy,” Reeser adds.
The Ohio State University Comprehensive Cancer Centerhttp://tinyurl.com/yczce7e2

Surprising discovery about how neurons talk to each other

Researchers at the University of Pittsburgh have uncovered the mechanism by which neurons keep up with the demands of repeatedly sending signals to other neurons. The new findings, made in fruit flies and mice, challenge the existing dogma about how neurons that release the chemical signal dopamine communicate, and may have important implications for many dopamine-related diseases, including schizophrenia, Parkinson’s disease and addiction.
Neurons communicate with one another by releasing chemicals called neurotransmitters, such as dopamine and glutamate, into the small space between two neurons that is known as a synapse. Inside neurons, neurotransmitters awaiting release are housed in small sacs called synaptic vesicles.
“Our findings demonstrate, for the first time, that neurons can change how much dopamine they release as a function of their overall activity. When this mechanism doesn’t work properly, it could lead to profound effects on health,” explained the study’s senior author Zachary Freyberg, M.D., Ph.D., who recently joined Pitt as an assistant professor of psychiatry and cell biology. Freyberg initiated the research while at Columbia University.
When the researchers triggered the dopamine neurons to fire, the neurons’ vesicles began to release dopamine as expected. But then the team noticed something surprising: additional content was loaded into the vesicles before they had the opportunity to empty. Subsequent experiments showed that this activity-induced vesicle loading was due to an increase in acidity levels inside the vesicles.
“Our findings were completely unexpected,” said Freyberg. “They contradict the existing dogma that a finite amount of chemical signal is loaded into a vesicle at any given time, and that vesicle acidity is fixed.”
The team then demonstrated that the increase in acidity was driven by a transport channel in the cell’s surface, which allowed an influx of negatively charged glutamate ions to enter the neuron, thus increasing its acidity. Genetically removing the transporter in fruit flies and mice made the animals less responsive to amphetamine, a drug that exerts its effect by stimulating dopamine release from neurons.
“In this case, glutamate is not acting as a neurotransmitter. Instead it is functioning primarily as a source of negative charge, which is being used by these vesicles in a really clever way to manipulate vesicle acidity and therefore change their dopamine content,” Freyberg said. “This calls into question the whole textbook model of vesicles as having fixed amounts of single neurotransmitters. It appears that these vesicles contain both dopamine and glutamate, and dynamically modify their content to match the conditions of the cell as needed.”
In the future, the team plans to look more closely at how increases in vesicle acidification affect health. A number of brain diseases are characterized by abnormal dopamine neuron signalling and altered levels of the neurotransmitter.
“Since we have demonstrated that the balance between glutamate and dopamine is important for controlling the amount of dopamine that a neuron releases, it stands to reason that an imbalance between the two neurotransmitters could be contributing to symptoms in these diseases,” said Freyberg.
University of Pittsburghhttp://tinyurl.com/y7laad5d

Mologic receives CE-mark approval for BVPro point-of-care diagnostic for bacterial vaginosis

Mologic Ltd, which develops personalized diagnostics, recently announced it has received CE-mark approval for BVPro®, a rapid point-of-care test for patients presenting with symptoms of vaginosis. The company also announced the commercial launch of the test, and that it has signed a 3-year distribution agreement with Hitado GmbH, a leading diagnostic supplier specializing in point-of-care products, and subsidiary of Sysmex, to enable distribution of BVPro throughout Germany.
Bacterial vaginosis (BV) is an abnormal vaginal condition that is characterized by vaginal discharge and results from an overgrowth of atypical bacteria in the vagina. Although not critical in its own right, BV is associated with serious medical complications such as post-operative infections, a greater susceptibility of contracting sexually transmitted infections (STIs) such as genital herpes and HIV, and also a higher risk of preterm birth in pregnant women.
BVPro is a simple, 15-minute, point-of-care diagnostic for professional use with patients presenting with symptoms of vaginosis. It detects sialidase enzyme activity from a vaginal swab sample which is a well-established clinical marker of bacterial vaginosis. The CE-marked diagnostic has been designed in the familiar lateral flow format (typical of pregnancy tests) so that the visual result is easy to interpret and enables a rapid, alternative to current methods, such as wet mount microscopic analysis and test tube-based colour-change assays, which often provide ambiguous results and are time consuming.
Mark Davis, co-founder and CEO at Mologic commented: “BVPro is another in Mologic’s pipeline of diagnostic tests to receive CE-mark approval. We are very pleased to be making such progress in bringing point-of-care tests to patients and by doing so, enabling the earlier treatment of a range of diseases, including sepsis, urinary tract infections and chronic obstructive pulmonary disease (COPD). Appointing Hitado to distribute our products in Germany represents a significant milestone for our company; we look forward to working with them as our first formal distributor and also others as we bring them on board in different geographies. www.mologic.co.uk/bvpro

Beckman Coulter highlights the first US IVD test delivering flow cytometric leukemia & lymphoma analysis in the routine clinical lab

ClearLLab reagents are the first to receive Food and Drug Administration (FDA) clearance (via the De Novo Process) to market them in the US.  They deliver the first preformulated, IVD antibody cocktails for leukemia and lymphoma immunophenotyping in the clinical lab. For clinical laboratories it means they no longer have to develop their own laboratory developed test (LDT), a technically demanding, manual, time-consuming, and potentially error-prone process. Previously, labs would have had to make and validate their own antibody cocktails.  ClearLLab reagents simplify and standardize the process.
 ‘FDA allows marketing of test to aid in the detection of certain leukemias and lymphomas’ with the FDA confirming that the test ‘provides consistent results to aid in the diagnoses of these serious cancers’.  The FDA evaluated data from a multi-site clinical study which compared panel results to alternative detection methods.
Dr Mario Koksch, Vice President and General Manager of Beckman Coulter’s Cytometry Business Unit said: “Flow cytometry is a powerful tool for the detailed and fast analysis of complex populations, with the technique becoming increasingly valuable to the clinical hematology laboratory. “Clearance to market the first IVD L&L reagents in the US has opened the door to the expansion globally of our clinical reagent and instrument portfolio.”  
ClearLLab reagents deliver fast and accurate qualitative identification of various hematolymphoid cell populations by immunophenotyping on the FC500 flow cytometer.  With the reliability of a standardized kit and protocols, the preformulated combinations offer LEAN-focused benefits which reduce manual preparation and validation time, accelerate sample preparation time, improve workflow, streamline lab inventory management and provide confidence in the accuracy and reliability of results.
As Dr Koksch added: “The routine use of ready-to-use ClearLLab reagents delivers greater efficiency and cost savings.  Preformulated antibody combinations enable the lab to avoid the potential errors of manual antibody cocktail preparation, with the reassurance of standardized reporting to international guidelines.”
ClearLLab reagents follow the 2006 Bethesda International Consensus Recommendations on the Flow Cytometric Immunophenotypic Analysis of Hematolymphoid Neoplasia. They are compatible with the World Health Organization (WHO) 2016-revised classification of myeloid neoplasms and acute leukemia.  WHO, in collaboration with the European Association for Hematopathology and the Society for Hematopathology, recently made important changes to the classification of these diseases. These included new criteria for the recognition of some previously described neoplasms as well as clarification and refinement of the defining criteria for others. www.beckman.com

Shimadzu has acquired AlsaChim, a specialist for high-quality analytical isotope-labelled standards

Shimadzu has joined forces with the France-based AlsaChim company specializing in stable isotope-labelled compounds, metabolites and pharmaceutical related substances. With immediate effect, Shimadzu Europe has acquired AlsaChim by 100%.The brand name will be kept for the future complemented by the subtitle “a Shimadzu Group Company”. Through this acquisition, Shimadzu will further develop and extend its activities in the clinical market which is one of the focus areas for the European Innovation Center (EUIC). The AlsaChim technology complements Shimadzu’s product and solution portfolio in the clinical market. Now, Shimadzu is able to enter the market with complete solutions consisting of hardware and software as well as application kits. Clients now benefit from one-stop solutions of complex analytical systems. www.shimadzu.eu/clinical

Siemens Healthineers has announced new strategic relationship in molecular testing with Fast-track Diagnostics

In an effort to provide customers with expanded, more comprehensive solutions for molecular testing, Siemens Healthineers has announced a new strategic relationship with Fast-track Diagnostics (FTD) that includes the addition of FTD’s broad range of CE-marked kits and multisyndromic panels to the menu of the Siemens Healthineers VERSANT kPCR Molecular System. The addition of FTD’s broad menu of kits and panels – which cover conditions from respiratory to gastroenteritis to central nervous system (CNS) and childhood infections – increases the breadth of Siemens Healthineers’ complete molecular testing solution, ensuring leading-edge performance from extraction through detection and increasing workflow efficiency for molecular labs of all sizes.
 “Over the past 24 months, Siemens Healthineers has made significant advancements in the delivery of molecular diagnostic applications and services,” says Fernando Beils, Head of Molecular Diagnostics, Siemens Healthineers. “We continue to strengthen and broaden our Molecular System by offering a comprehensive, scalable solution for accurate diagnosis and monitoring to our customers worldwide through our alliance with Fast-track Diagnostics.”
The VERSANT kPCR Molecular System, an established market player in molecular testing for HIV and Hepatitis, will now feature over 75 assays, consolidating testing for the infectious disease spectrum in a single molecular ecosystem.
“The VERSANT kPCR Molecular System is perfectly suited to our wide range of assays, which means laboratories can now diagnose nearly any infectious disease in one workflow,” says Bill Carman, CEO of Fast-track Diagnostics.
In offering customers the option of a single, consolidated system with a broad menu and virtually open platform, Siemens Healthineers enables healthcare providers to meet their current challenges, especially as an increasing push towards a value-based healthcare philosophy relies heavily on increased productivity and streamlined workflows. With this in mind, Siemens Healthineers has made its growth and enhancement within the molecular diagnostics space a key priority for its business strategy moving forward. www.siemens.com/healthineerswww.fast-trackdiagnostics.com

DiaSys signs cooperation agreement with Tosoh Bioscience

DiaSys Diagnostic Systems GmbH has established a future-oriented cooperation with Tosoh Bioscience. The aim of this cooperation is to consolidate clinical chemistry (DiaSys BioMajesty® JCA-BM6010/C), immunoassays (Tosoh AIA-CL® 1200), hematology and coagulation analysis either simply with a middleware (EVOLINE® Manager; TOSOH) or fully automated with a track system. The conveyor system (EVOLINE®; Tosoh) offers the option to combine the DiaSys clinical chemistry analyser BioMajesty® JCA-BM6010/C with a wide range of analysers from different vendors. This partnership addresses the common trend of consolidation which aims to provide tailor-made solutions for individual customer needs.   www.diasys-diagnostics.com

Thermo Fisher Scientific offers preview of the world’s first fully integrated LC-MS/MS Clinical Analyser

Following the presentation to the European market at EuroMedLab in Athens and to the US market at AACC 2017 of the Thermo Scientific Cascadion SM Clinical Analyser bringing together the ease of use of clinical analysers with the selectivity and sensitivity of liquid chromatography-tandem mass spectrometry (LC-MS/MS), the company will now seek CE marking  followed by FDA approval.
“This is a marvelous development, and it is really quite outstanding. It will fulfill the needs of many laboratories,” said Professor Brian Keevil, consultant clinical scientist and head of the Clinical Biochemistry Department, University Hospital of South Manchester NHS Foundation Trust, UK, after viewing a demonstration during EuroMedLab 2017.
The Cascadion system was designed and built using Thermo Fisher products and technologies combined with its industry-leading expertise in mass spectrometry.  Featuring turnkey operation, the Cascadion analyser is designed to be used by laboratory staff with no specialized training.
James Nichols, PhD, medical director, Chemistry and Point of Care Testing, Vanderbilt University Medical Center, added, “For much of what we do in terms of chromatography and mass spectrometry, we need very highly skilled and experienced medical technologists. The Cascadion analyser is relatively maintenance-free and because it includes specially designed reagent kits, there is not a lot of interaction required with the technology.”
After previewing the Cascadion analyser, Michael Vogeser, senior physician and professor of laboratory medicine, University Hospital of Munich, stated “About 70% of all physician’s decisions are based on laboratory tests so the impact on laboratory testing is huge and this completely new technological approach is of enormous value to mankind.” www.thermofisher.com/Cascadion