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As analytical instrumentation (gas- and liquid-chromatographs coupled with mass spectrometers) increase in sensitivity and speed, forensic scientists may find themselves still hindered by the process of preparing samples (blood, urine, etc.) for analysis and seeking more efficient approaches.
In an article in WIRES Forensic Science, researchers from Boston University School of Medicine’s (BUSM) Biomedical Forensic Sciences program, provide an overview of sample preparation techniques and information on routine sample types that may be encountered in forensic toxicology cases.
Forensic toxicology encompasses a large variety of scenarios including drug-facilitated crimes, understanding the role alcohol or other drugs may have played in an individuals’ deaths, as well as complex polydrug use in driving under the influence cases. As variable as the cases, so too are the composition of biological matrices in addition to how to identify drugs or other compounds in biological samples. This has provided a vast array of sample preparation approaches that scientist have in their ‘toolbox.’ “Our work highlights the variability in sample types that toxicological analysis encompasses as well as vast array of sample preparation techniques that are currently available,” said corresponding author Sabra R. Botch-Jones, MS, MS, MA, assistant professor of anatomy and neurobiology at BUSM.
According to the researchers, the choice of biological matrix is dependent on the anticipated answer the toxicologist is trying to get. Is suspected drug use recent? If so, a blood or oral fluid sample may be best to assess the drugs present and how much is in the sample. In cases of driving while impaired, it is essential to obtain a sample that is appropriate to assess if the drug effected the driver’s ability to operate the vehicle safely. When drugs are used, the human body will break down, or metabolize, the drug and eventually excrete it. Drugs and their metabolites may be excreted or removed from the body over a course of hours and even days. For example, if there has been a delay in obtaining a sample or reporting a crime in which drugs are suspected, a urine sample may be best. If extensive amounts of time have elapsed, a hair sample may be a viable option to determine exposure to a suspected drug or other compound.
As with the instrumentation, sample preparation tools have also advanced over time. Solid phase and, more recently supported liquid extraction, allows the unwanted materials in the biological sample to be retained on a solid surface composed of natural materials such as silica or diatomaceous earth. These extraction tools can provide clean extracts containing the drugs of interest and help to recover a large variety of drugs, which can aid in laboratory efficiency when dealing with poly-drug cases.
“Researchers have a number of biological samples to choose from when trying to identify what substances may be in the human body, however it important that they choose the right one to help answer their research question. As equally important is the choice of how to prepare the sample for analysis. This work provides an overview of the routine biological samples, their components, and ways to process them for downstream analysis,” explains Botch-Jones.
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Most kids dream of growing up to be astronauts; but the downside of spending extended amounts of time in low gravity is that astronauts’ muscles tend to shrink and weaken through disuse. Now, researchers from Japan have identified a protein that affects how muscles respond to space flight.
In a study published in June 2021 in Communications Biology, researchers from the University of Tsukuba have revealed that nuclear factor E2-related factor 2, or NRF2, helps keep muscles from becoming weak in low gravity.
Muscle atrophy, or weakening, is a common feature of disease and aging. It can also occur after a long period of inactivity, such as during space flight, when astronauts don’t need to use their muscles as much as they do on Earth to support their weight or move around. When muscles atrophy in space, they not only decrease in size, but they also tend to lose a type of fiber called “slow-twitch” and gain more fibers called “fast-twitch.”
“This conversion from slow- to fast-twitch muscle fibers is closely associated with an increase in oxidative stress,” explains Professor Satoru Takahashi, the senior author of the study. “Thus, we expected that removing factors that protect against oxidative stress would accelerate muscle atrophy under microgravity conditions.”
To explore this, the researchers deleted the gene encoding NRF2, which helps controls the body’s response to oxidative stress, in mice. The mice were then sent to live on the International Space Station for a month. When the mice returned, the researchers compared their calf muscles with those from mice who had spent the same month on Earth.
“We were surprised to find that the Nrf2-knockout mice did not lose any more muscle mass than the control mice under a microgravity environment,” says Professor Takahashi. “However, they did show a significantly accelerated rate of slow-to-fast fiber type transition.”
In addition to this change in muscle composition, there were also noticeable changes in the way that the muscle tissue used energy and nutrients. This shift in energy metabolism is a common feature of fiber type transition.
“Our findings suggest that NFR2 alters skeletal muscle composition during space flight by regulating oxidative and metabolic responses,” states Professor Takahashi.
Given this newly discovered role for NFR2, finding treatments that target this protein could be useful for helping prevent muscle changes in astronauts during space flight. Targeting NFR2 could also be a promising avenue for addressing muscle wasting in diseases like cancer or during the aging process.
The article, “Nuclear factor E2-related factor 2 (NRF2) deficiency accelerates fast fibre type transition in soleus muscle during space flight,” was published in Communications Biology.
This work was supported by a Grant-in-Aid for the Japan Aerospace Exploration Agency (14YPTK-005512; S.T.), and a Grant-in-Aid for Scientific Research on Innovative Areas from MEXT (18H04965; S.T.). The authors declare no competing interests.
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Researchers have made significant advancements in correlating aberrations in specific brain circuits with neuropsychiatric conditions like depression. However, it remains difficult to prove that damage to these circuits causes the symptoms themselves and that targeting them with therapeutics could help patients. By integrating brain lesion datasets with data on how two treatments — deep brain stimulation (DBS) and transcranial magnetic stimulation (TMS) — influence neuropsychiatric disorders, researchers from Brigham and Women’s Hospital and collaborators developed a new brain mapping approach that may help clarify the cause of a variety of neuropsychiatric conditions and identify promising stimulation sites to target therapeutically. Findings are published in Nature Human Behavior.
“This is a new technique that uses existing data on patients with brain damage to develop new treatment targets for real-world patients with similar symptoms,” said the paper’s corresponding author, Shan Siddiqi, MD, of the Center for Brain Circuit Therapeutics at the Brigham. “In principle, this should open the floodgates for researchers to study any stroke- or brain-injury-associated symptom to find a new treatment target for people who developed the same symptom without brain damage.”
The researchers developed their approach using data on depression and Parkinson’s disease, both of which are already associated with well-defined brain lesions and are commonly treated with DBS and TMS. They combined the location and connectivity of 461 brain lesions, 101 DBS sites, and 151 TMS sites, and compared patients who developed depression, patients who had improvement in depression, and patients who had no change in mood. Using this approach, they identified a brain circuit that is an effective therapeutic target for both invasive and noninvasive brain stimulation treatments. The study also indicates that brain stimulation outcomes vary not according to the technique used — DBS versus TMS — but according to the circuit that is targeted.
The researchers subsequently used their approach with Parkinson’s disease data. Combining data on 29 lesions and 95 stimulation sites for tremors and rigidity, they showed that lesions associated with the motor symptoms of Parkinson’s disease are connected to the same circuits as the stimulation sites that that relieve those symptoms.
The researchers are now working to refine circuit maps for other neuropsychiatric conditions such as anxiety disorders, post-traumatic stress disorder, mania, hallucinations, and movement disorders. Clinical trials are still necessary to determine whether physicians can relieve patient symptoms by targeting the brain circuits identified through the new approach. The authors of the study are currently conducting a trial to confirm the distinct TMS targets that they recently identified for depression and anxiety.
“Now that we have concrete evidence that lesions map to treatment targets, we can design more clinical trials to generate new treatments,” Siddiqi said. “This approach gives us highly rigorous hypotheses about treatment targets. When we don’t know much about the brain circuitry of a particular disorder, our study shows how to find the answer to that question and turn it into new treatment targets.”
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Lung cancer has the highest mortality rate of all cancers, and treatment options are extremely limited, especially for patients with oncogenic mutations in the KRAS gene. A great deal of hope was invested in the licensing of immune checkpoint inhibitors, but the reality is that some patients respond very well to this treatment while it is completely ineffective in others. In a paper just published in Science Translational Medicine, a MedUni Vienna research group led by Herwig Moll (Center for Physiology and Pharmacology) identified a potential marker for the success of immunotherapy in lung cancer patients and explained the underlying molecular processes.
K-Ras it is a monomeric G protein that plays a key role in the growth of malignant tumours. KRAS-mutated lung carcinomas frequently occur in chronically inflamed lungs, particularly in heavy smokers. The inflammatory processes promote the growth of cancer cells. The research group has now shown that the expression of the highly anti-inflammatory protein A20, formed in the body itself, is often very low in these malignant cells and that there is a direct correlation between a patient’s life expectancy and the expression of this protein. Moll explains: “Both in humans and in the animal model, the loss of A20 leads to downgraded immune surveillance of cancer cells. Cancer cells with low levels of A20 are able to escape detection by the immune system.” This results in significantly faster tumour growth.
During the course of this study, which was co-funded by MedUni Vienna’s Cancer Research Initiative and associated with the Comprehensive Cancer Center Vienna, the research team discovered that it is primarily an enhanced sensitivity of the cancer cells to the immunomodulatory cytokine interferon gamma that is responsible for this. Moreover, tumour cells with downregulated A20 responded particularly well to immune checkpoint inhibitors, in the same way as patients suffering from melanoma (skin cancer) with a similar gene expression structure.
“In A20 we have discovered a previously unknown tumour suppressor in lung cancer, the loss of which as an immune checkpoint contributes to the development of this malignant disease,” explains co-author Emilio Casanova from the Institute of Pharmacology. Since patients with low A20 expression have few tumour-fighting immune cells and so, in the advanced stage, express little of the important immune checkpoint molecule PD-L1, these patients could be excluded from immunotherapies directed against PD-L1. Indeed, the strength of expression of this molecule is currently regarded as an aid for deciding whether or not they should be treated with immune checkpoint inhibitors. “Based on our results and the data available from melanoma patients, we are convinced that we have identified a group of lung cancer patients who would really benefit from this immunotherapy. Exclusion from such treatment would significantly reduce the survival rate of such patients.”
In a further study, the researchers want to find out whether it is possible to manipulate the expression of A20 in the cancer cells, in order to intensify the effect of immunotherapies. “However, smoking is still the most easily avoided risk factor for lung cancer. We must therefore support laws to protect the general public from inhaling harmful smoke, while at the same time appealing to people’s personal responsibility to refrain from smoking altogether,” says Moll. According to the MedUni Vienna experts, it is nevertheless important to continue to investigate new therapeutic approaches to improve the quality-of-life and chances of survival of those affected.
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Researchers at The University of Queensland, working to gain a better understanding of how brain cells work, have discovered the underlying mechanism of a rare genetic mutation that can cause epilepsy.
Dr Victor Anggono from UQ’s Queensland Brain Institute said his team made the ground-breaking findings while researching nerve cell communications, which are an important process in normal brain function.
”We’re both excited and astounded to make such an important contribution to the field of cellular and molecular neuroscience,” Dr Anggono said.
He stressed that the mutation was extremely rare, with only one reported case in the world to date.
Dr Anggono’s team studied protein structures, called receptors, that are attached to cell surfaces to make the discovery.
”It turns out that this particular mutation causes receptors in brain cells to behave differently, resulting in an imbalance in brain cell communication — and that can lead to disorders,” he said.

A joint research group from KAIST and Institut Pasteur Korea has identified repurposed drugs for COVID-19 treatment through virtual screening and cell-based assays. The research team suggested the strategy for virtual screening with greatly reduced false positives by incorporating pre-docking filtering based on shape similarity and post-docking filtering based on interaction similarity. This strategy will help develop therapeutic medications for COVID-19 and other antiviral diseases more rapidly. This study was reported at the Proceedings of the National Academy of Sciences of the United States of America (PNAS).
Researchers screened 6,218 drugs from a collection of FDA-approved drugs or those under clinical trial and identified 38 potential repurposed drugs for COVID-19 with this strategy. Among them, seven compounds inhibited SARS-CoV-2 replication in Vero cells. Three of these drugs, emodin, omipalisib, and tipifarnib, showed anti-SARS-CoV-2 activity in human lung cells, Calu-3.
Drug repurposing is a practical strategy for developing antiviral drugs in a short period of time, especially during a global pandemic. In many instances, drug repurposing starts with the virtual screening of approved drugs. However, the actual hit rate of virtual screening is low and most of the predicted drug candidates are false positives.
The research group developed effective filtering algorithms before and after the docking simulations to improve the hit rates. In the pre-docking filtering process, compounds with similar shapes to the known active compounds for each target protein were selected and used for docking simulations. In the post-docking filtering process, the chemicals identified through their docking simulations were evaluated considering the docking energy and the similarity of the protein-ligand interactions with the known active compounds.
The experimental results showed that the virtual screening strategy reached a high hit rate of 18.4%, leading to the identification of seven potential drugs out of the 38 drugs initially selected.
“We plan to conduct further preclinical trials for optimizing drug concentrations as one of the three candidates didn’t resolve the toxicity issues in preclinical trials,” said Woo Dae Jang, one of the researchers from KAIST.
“The most important part of this research is that we developed a platform technology that can rapidly identify novel compounds for COVID-19 treatment. If we use this technology, we will be able to quickly respond to new infectious diseases as well as variants of the coronavirus,” said Distinguished Professor Sang Yup Lee.
This work was supported by the KAIST Mobile Clinic Module Project funded by the Ministry of Science and ICT (MSIT) and the National Research Foundation of Korea (NRF). The National Culture Collection for Pathogens in Korea provided the SARS-CoV-2 (NCCP43326).
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The states, including Massachusetts and New York, agreed to drop opposition to the bankruptcy organization plan of the company, the maker of OxyContin.Fifteen states have reached an agreement with Purdue Pharma, the maker of the prescription painkiller OxyContin, that would pave the way toward a $4.5 billion settlement of thousands of opioid cases.The states decided late Wednesday to drop their opposition to Purdue’s bankruptcy reorganization plan, in exchange for a release of millions of documents and an additional $50 million from members of the Sackler family, the company’s owners.The agreement was contained in a late-night filing by a mediator in U.S. Bankruptcy Court in White Plains, N.Y.The settlement extracts concessions that will be added to a comprehensive proposal now being voted upon by more than 3,000 plaintiffs, including cities, counties, tribes and states, who sought to hold Purdue and its owners responsible for their role in the opioid epidemic, during which more than 500,000 Americans have died from overdoses of prescription and illegal opioids.Trials against other opioid manufacturers and drug distributors are underway.Nearly two years ago, the Sacklers had proposed paying $3 billion in cash. Both the company and family members had resisted releasing the full trove of documents, including hundreds of thousands of work emails and communications with attorneys, reaching back decades. According to last night’s filing, Purdue and the Sacklers will now release some 33 million documents, and the money has risen to $4.5 billion, plus an additional $225 million in a civil settlement with the Department of Justice.According to spokesmen, two branches of the Sackler family noted that the settlement included no finding of liability or wrongdoing. In a statement, they said: “This resolution to the mediation is an important step toward providing substantial resources for people and communities in need. The Sackler family hopes these funds will help achieve that goal.”The Sacklers will have nine years to make payments, but the new agreement includes an enhanced schedule.If Judge Robert Drain, who is presiding over the bankruptcy proceedings, certifies the plan after an August hearing, as is now widely expected, both the family and the company would be shielded from further opioid-related lawsuits.Maura Healey, the attorney general of Massachusetts, who was the first to sue individual Sacklers, said, “While I know this resolution does not bring back loved ones or undo the evil of what the Sacklers did, forcing them to turn over their secrets by providing all the documents, forcing them to repay billions, forcing the Sacklers out of the opioid business, and shutting down Purdue will help stop anything like this from ever happening again.”Another official in the pursuit of the Sacklers was Letitia James, the attorney general of New York.“For nearly two years, since Purdue Pharma declared bankruptcy, the company and the Sackler family have used every delay tactic possible and misused the courts — all in an effort to shield their misconduct,” she said. “While this deal is not perfect, we are delivering $4.5 billion into communities ravaged by opioids on an accelerated timetable and it gets one of the nation’s most harmful drug dealers out of the opioid business once and for all. ”Nine states and the District of Columbia continue to oppose the agreement. “While some progress has been made — especially around the public document depository — this plan is far from justice,” said William Tong, the attorney general of Connecticut. “Purdue and the Sacklers have misused this bankruptcy to protect their vast wealth and evade consequences for their callous misconduct. This deal alarmingly allows the Sacklers to still walk away with their personal wealth intact, and now allows funds already intended for charity to be included in this deal. We are evaluating all options to continue to fight this bankruptcy plan until all viable options are exhausted.”

The vast majority of electronic health record (EHR) alerts attempting to reduce the prescribing of high-risk medications linked to dementia in older adults went unread in a study led by research scientists from Regenstrief Institute, Purdue University and Indiana University School of Medicine. The goal of the intervention was to facilitate the deprescribing of anticholinergics through both provider and patient-based alerts, however, engagement with the alerts was so low, the study team was unable to conclude if this approach could be an effective method.
Anticholinergics are drugs which affect the brain by blocking acetylcholine, a nervous system neurotransmitter that influences memory, alertness and planning skills. They are linked to dementia and prescribed for many conditions common in older adults including depression, urinary incontinence, irritable bowel syndrome and Parkinson’s disease. These medications are used by approximately one in four older adults each year, and nearly half of older adults have used this type of medication at least once in a five-year period.
Many medical groups have come out in support of deprescribing anticholinergics, but it is challenging to execute in an already busy primary care environment.
“Deprescribing is very complex and rarely prioritized over common medical problems during visits with primary care providers,” said study lead author Noll Campbell, PharmD, M.S., research scientist at the IU Center for Aging Research at Regenstrief and assistant professor of pharmacy at Purdue University College of Pharmacy. “In this study, we used principles of behavioral economics in the design of EHR nudges directed at both providers and patients to promote the deprescribing of high-risk anticholinergic medications. However, very few of the alerts were viewed by either recipient, so we are now evaluating how we can change or improve this approach.”
In this study, an alert let the provider know that the patient had high-risk anticholinergic medications prescribed in the medical record and offered alternatives to those medications. Alerts also prompted staff to play a video providing education about the medicines and modeling a discussion that led to a change in prescription for patients who were prescribed one of the target medications.
The research team conducted the cluster randomized trial in Eskenazi Health clinics and compared the medication records to the previous year to see if there were any changes. They found there were no significant differences in deprescribing between the control group and the intervention group.
During the course of the study, 85 percent of alerts to providers and 95 percent of alerts to medical assistants went unread, so study authors cannot conclude that priming patients and providers for the discussion is not a feasible strategy, only that the methods used in this study were not successful in reaching the target recipients.
“One option going forward is to experiment with different design approaches in EMR-based nudges,” said Dr. Campbell. “Alternatively, a shift towards human-based interventions that can manage the complexity of deprescribing activities may be more effective at deprescribing high-risk anticholinergic medications. While we pursue the goal of understanding clinical implications, we are also cognizant of the scalability of interventions if there is clinical benefit realized by reducing these high-risk medications.”
Dr. Campbell and his colleagues at Regenstrief are currently conducting a clinical trial designed to determine if stopping anticholinergic medications results in sustained improvements in cognition. This trial involves clinical pharmacists working with physicians and patients to switch to safer medicines.
Another study at the IU Center for Aging Research at Regenstrief is testing an app called BrainSafe, which provides information on anticholinergics to patients with the goal of leading them to initiate a deprescribing conversation.
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Using single cell technology, a new study sheds light on the significance of genetic risk factors for, and the diversity of cells involved in, the development of coronary artery disease. The researchers analysed human atherosclerotic lesions to map the chromatin accessibility of more than 7,000 cells. The chromatin accessibility is known to reflect active regions and genes in the genome. The findings were published in Circulation Research.
Genome-wide association studies of the human genome have identified over 200 loci associated with coronary artery disease. More than 90% of them are located outside protein-coding genes, in so called cis-regulatory elements, whose significance in the pathogenesis of coronary artery disease remains unclear.
Previous research has demonstrated that the development of coronary artery disease involves a variety of different cells and their subtypes. The now-published study is the first to use single cell technology to map epigenetic changes in these cells. The researchers used the ATAC-seq sequencing method to discover the nuclear chromatin structure of endothelial cells and smooth muscle cells, as well as immune system monocytes, macrophages, NK/T and B cells, providing a unique resource to study the cell-type specific activity of the cis-regulatory elements in the disease affected vessel wall.
The study demonstrated that genetic risk variants associated with coronary artery disease are particularly enriched in cis-regulatory elements specific to endothelial and smooth muscle cells, indicating that these cells play a significant role in transmitting susceptibility to the disease. Based on chromatin accessibility mapping and gene expression data, the researchers were able to identify putative target genes for approximately 30% of all known loci associated with coronary artery disease. In addition, the researchers performed genome-wide experimental fine-mapping of the variants, allowing them to identify potential causal single-nucleotide polymorphisms and the associated target gene for over 30 loci that have been linked to coronary artery disease. The study also presented a number of examples of how the chromatin accessibility and gene expression data can be used to predict target cells via which the function of the genetic changes associated with the disease is transmitted in the tissue. This is a significant step forward that helps to understand the real functional significance of risk variants in the pathophysiology of coronary artery disease. In the future, this information can be used to develop more effective, safer and more individualised treatments for coronary artery disease.
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Tooth loss is a risk factor for cognitive impairment and dementia — and with each tooth lost, the risk of cognitive decline grows, according to a new analysis led by researchers at NYU Rory Meyers College of Nursing and published in JAMDA: The Journal of Post-Acute and Long-Term Care Medicine. However, this risk was not significant among older adults with dentures, suggesting that timely treatment with dentures may protect against cognitive decline.
About one in six adults aged 65 or older have lost all of their teeth, according to the Centers for Disease Control and Prevention. Prior studies show a connection between tooth loss and diminished cognitive function, with researchers offering a range of possible explanations for this link. For one, missing teeth can lead to difficulty chewing, which may contribute to nutritional deficiencies or promote changes in the brain. A growing body of research also points to a connection between gum disease — a leading cause of tooth loss — and cognitive decline. In addition, tooth loss may reflect life-long socioeconomic disadvantages that are also risk factors for cognitive decline.
“Given the staggering number of people diagnosed with Alzheimer’s disease and dementia each year, and the opportunity to improve oral health across the lifespan, it’s important to gain a deeper understanding of the connection between poor oral health and cognitive decline,” said Bei Wu, PhD, Dean’s Professor in Global Health at NYU Rory Meyers College of Nursing and co-director of the NYU Aging Incubator, as well as the study’s senior author.
Wu and her colleagues conducted a meta-analysis using longitudinal studies of tooth loss and cognitive impairment. The 14 studies included in their analysis involved a total of 34,074 adults and 4,689 cases of people with diminished cognitive function.
The researchers found that adults with more tooth loss had a 1.48 times higher risk of developing cognitive impairment and 1.28 times higher risk of being diagnosed with dementia, even after controlling for other factors.
However, adults missing teeth were more likely to have cognitive impairment if they did not have dentures (23.8 percent) compared to those with dentures (16.9 percent); a further analysis revealed that the association between tooth loss and cognitive impairment was not significant when participants had dentures.
The researchers also conducted an analysis using a subset of eight studies to determine if there was a “dose-response” association between tooth loss and cognitive impairment — in other words, if a greater number of missing teeth was linked to a higher risk for cognitive decline. Their findings confirmed this relationship: each additional missing tooth was associated with a 1.4 percent increased risk of cognitive impairment and 1.1 percent increased risk of being diagnosed with dementia.
“This ‘dose-response’ relationship between the number of missing teeth and risk of diminished cognitive function substantially strengthens the evidence linking tooth loss to cognitive impairment, and provides some evidence that tooth loss may predict cognitive decline,” said Xiang Qi, a doctoral candidate from NYU Meyers.
“Our findings underscore the importance of maintaining good oral health and its role in helping to preserve cognitive function,” said Wu.
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