Monday, April 15, 2024

 

Aston University to help Saudi Arabia turn waste into energy


ASTON UNIVERSITY
Aston University to help Saudi Arabia turn waste into energy 

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KEW TECHNOLOGY’S SUSTAINABLE ENERGY CENTRE IN WEDNESBURY, UK

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CREDIT: ASTON UNIVERSITY




  • Energy will help power new cities in the desert
  • Aston University is in talks about converting waste products into vital energy
  • Its Energy and Bioproducts Institute is experienced in the waste-to-energy sector through global collaborations.

Aston University researchers are to help turn waste into energy to power new cities in the desert.

The University has started talks with experts from Saudi Arabia, including those who are building two sustainable cities in the desert, called NEOM and The Line.

They are to collaborate with Aston University and its Energy and Bioproducts Research Institute (EBRI) to explore how they can convert waste products into vital energy. 

The scientists and engineers are to apply their expertise to help Saudi Arabia create technology to convert discarded matter into a source of energy and other innovations such as using date palm waste to transform desert sand to allow it to retain water and grow crops.

Aston University also hosted a two-day conference in March to discuss how to develop and apply the technology.
The event is a key element of the UK-KSA Waste2Energy project supported by the Foreign, Commonwealth and Development Office under the Gulf Strategy Fund (GSF) programme and is led by senior lecturer in mechanical, biomedical and design engineering Dr Muhammad Imran.  

More than 70 delegates attended the conference, including representatives from King Abdulaziz City for Science and Technology (KACST), King AbdulAziz University, The National Research and Development Center for Sustainable Agriculture and the Saudi Investment Recycling Company (SIRC).

Professor Patricia Thornley, director of Energy & Bioproducts Research Institute, said: “The delegation chose to collaborate with and visit EBRI because we have common research goals, but some complementarity facilities and skills. We are looking forward to working together to develop some the shared priorities we have identified.”

Tim Miller, EBRI director of engagement, added: “Aston University has extensive engagement in the waste-to-energy sector through substantial industrial and academic collaborations globally. Advancements made by institutes like EBRI in waste-to-energy technologies are continually contributing to sustainable energy development.”

“The meeting provided an insightful overview of the project, emphasising the significant opportunities it offers to UK industries and academia for funding, collaboration and PhD opportunities.

“Our special appreciation is extended to Naif Makki from the Ministry of Energy, Saudi Arabia and his colleagues for their valuable participation.”

The event ended with a tour of the EBRI lab and biochar demonstrator plant and a visit to Kew Technology’s Sustainable Energy Centre in Wednesbury.

 

Air pollution data could be used to plan better transit routes


Researchers from the University of São Paulo are producing maps that show areas of the city of São Paulo with the highest concentration of air pollutants; the results of the studies were presented in the United States during FAPESP Week Illinois



FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO

Air pollution data could be used to plan better transit routes 

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TERESA CÓRDOVA, MARIANA GIANNOTTI AND THIAGO NOGUEIRA. RESERCHERS ARE PRODUCING MAPS THAT SHOW THE AREAS OF THE CITY WITH THE HIGHEST POLLUTION LEVELS AND THE PERIODS OF PEAK EMISSIONS.

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CREDIT: ELTON ALISSON




 In addition to traffic information, in the not-too-distant future it will be possible to decide the best route and time to travel by car in cities like São Paulo – the capital of the state of the same name and Brazil’s largest metropolis – based on air quality data. Through studies supported by FAPESP, researchers from the Institute of Astronomy, Geophysics and Atmospheric Sciences (IAG) and the School of Public Health (FSP) of the University of São Paulo (USP) are producing maps that show the areas of the city with the highest pollution levels and the periods of peak emissions.

Some of the results of the projects were presented during a panel discussion on opportunities for studies on smart cities, held on April 9th at the opening of FAPESP Week Illinois, in Chicago (United States).

Organized by FAPESP in partnership with the University of Illinois System (UIS) – composed of the universities of Illinois in Chicago, Urbana-Champaign and Springfield – at the headquarters of the Discovery Partners Institute in Chicago, the meeting aims to create opportunities for scientific and technological cooperation between researchers from the state of São Paulo and the Midwest of the United States.

The event brings together researchers from universities and research institutions in the state of São Paulo, from the UIS, from the Great Lakes region of North America – located on the border between the United States and Canada – and from partner institutions in Canada and Mexico.

Participants’ areas of expertise include health and medicine, smart agriculture, climate, bioenergy, investing in democratic institutions, and smart cities.

“Smart cities use innovative information technologies to collect data that are used to build and operate interconnected urban systems to improve efficiency and enhance sustainability and resilience,” said Teresa Córdova, director of the Great Cities Institute of the University of Illinois in Chicago.

According to the researcher, some key technologies that are driving the development of solutions to make cities in different parts of the world smarter are fiber optics, radio signal sensors, and cell phone frequencies.

In São Paulo, for example, researchers from IAG and FSP-USP are using portable equipment such as laser particle counters and GPS devices installed in cars to measure and obtain geo-referenced data on the concentration of particulate matter to which drivers and passengers are exposed in the city on different routes and at different times of day.

"Based on the data collected during these studies, we were able to make maps and identify which regions of the city have the highest concentration of pollutants," Thiago Nogueira, professor at FSP-USP, told Agência FAPESP.

The researchers conducted a study assessing exposure to air pollutants during commutes by different modes of transportation in São Paulo – including car, bus, subway and bicycle – and in different regions of the city. The results showed that residents in the western region of the capital are exposed to higher concentrations of an air pollutant called black carbon. The regions with the best air quality were those with the highest concentration of green spaces. On the other hand, the regions with intermediate air quality had more high-rise buildings.

“We’ve seen that high-rise buildings have a negative effect on the dispersion of air pollution,” said Nogueira.

As part of an international project, the researchers also measured the levels of exposure to particulate matter in ten cities around the world, including São Paulo. The results of the study showed that off-peak exposure was 40% lower in the morning and 91% lower at night. The highest levels of exposure to high-concentrated particulate matter were recorded in situations where the car windows were open.

Another finding of the study was that bus drivers and passengers in São Paulo are exposed to higher levels of air pollutants than car and subway users. However, the levels of atmospheric exposure in São Paulo’s transport system were lower than those recorded in the other nine participating cities, located in Africa, Asia and South America.

“Despite the culture that São Paulo is a very polluted city, in comparison with other countries we observed that the levels of exposure to particulate matter in the capital are lower,” said Nogueira.

Some of the reasons for this difference are that the fuels used to power vehicles in Brazil are cleaner than those used in the countries of the other nine participating cities. In addition, Brazil’s energy matrix has a larger share of renewable sources, Nogueira explained.

“In Brazil, we also have Proconve [Health Program for the Control of Air Pollution from Motor Vehicles], which at every stage has established the requirement to use cleaner fuels and more efficient engines in the country,” Nogueira said.

Unequal access

Currently, 70% of the world’s greenhouse gas (GHG) emissions come from cities, almost half of which come from urban transportation, mostly from private cars, said Mariana Giannotti, professor at the Polytechnic School of USP (POLI-USP) and technology transfer coordinator at the Center for Metropolitan Studies (CEM) – a FAPESP Research, Innovation and Dissemination Center (RIDC). “In this sense, public transport represents an excellent strategy to address issues related to sustainability,” said the researcher during a lecture at the event.

Through a study conducted within the framework of the CEM, the researcher and collaborators compared access to public transport in São Paulo, New York, and London, based on users’ travel time and ticket costs. The results show that New York has a public transport integration policy that is fair and helps to reduce inequality in access to public transport. The situation in São Paulo is different, according to Giannotti.

“In São Paulo, inequality is being exacerbated by the way public transport users are charged. Normally, people in the city spend on average between 20% and 40% of their income on it, while New Yorkers spend between 5% and 10%,” she compared.

The biggest users of buses in São Paulo are people from lower economic classes, especially blacks, the researcher said.

“Rich people, who live in the best places in São Paulo, have access to better quality public transportation and are paying less,” she compared.

 

Industry gifts may influence which cardiac device is used in common lifesaving procedure



UNIVERSITY OF PITTSBURGH


PITTSBURGH, April 10, 2024 — Physicians who received payments from the manufacturer of a specific set of cardiac devices are more likely to use those devices, often during a common procedure to unblock coronary arteries, according to a research letter published today in JAMA authored by researchers from the University of Pittsburgh, University of California, San Francisco, Yale University and the University of Connecticut.

The devices, known as the intravascular microaxial left ventricular assist devices (LVADs), were developed to provide cardiac support, often during percutaneous coronary intervention (PCI) procedures, and have been scrutinized in recent years for concerns about the evidence supporting their safety and effectiveness. The Food and Drug Administration issued a correction on instructions for LVADs last month. The research letter raises questions about how industry payments influence the use of medical devices, which can include high-risk procedures as well as high costs.

Using the national disclosure program Open Payments, the authors examined marketing-related payments from the LVAD manufacturer to cardiologists from 2016 to 2018, then linked these data to Medicare claims to determine which devices were subsequently used in beneficiaries. The team found that physicians who received payments from the manufacturer of LVADs were approximately twice as likely to use the device that year as well as in the following year, suggesting a lasting effect.

Senior author Timothy Anderson, M.D., M.A.S., a primary care physician, health services researcher and assistant professor of medicine at Pitt, explained that their findings align with previous studies showing that even small marketing payments may influence clinical care, but that most prior studies focus on prescription drugs. This study is one of the first to expand to medical devices and raises concern given that intravascular microaxial LVADs are more expensive but not shown to be more safe or effective than alternatives.

“These marketing relationships are very different from research partnerships between academia and industry, which are often beneficial in developing new innovations,” said Anderson. “If free meals and gifts to doctors might influence what device is used during a cardiac emergency, it is time to rethink whether these relationships are acceptable.”

Other authors on the study were Sanket Dhruva, M.D., M.H.S., of UCSF, Joseph S. Ross, M.D., M.H.S., of Yale; Michael Steinman, M.D., and Siqi Gan, M.P.H., of UCSF; and Sruthi Muluk, M.D., of University of Connecticut.

This research was supported by Veterans Affairs Health Services Research & Development (#1IK2HX003357) and National Institutes of Health/National Institute on Aging (#K76AG074878, #2K24AG049057, #P30 AG044281).

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About the University of Pittsburgh School of Medicine

As one of the nation’s leading academic centers for biomedical research, the University of Pittsburgh School of Medicine integrates advanced technology with basic science across a broad range of disciplines in a continuous quest to harness the power of new knowledge and improve the human condition. Driven mainly by the School of Medicine and its affiliates, Pitt has ranked among the top recipients of funding from the National Institutes of Health since 1998. In rankings released by the National Science Foundation, Pitt is in the upper echelon of all American universities in total federal science and engineering research and development support.

Likewise, the School of Medicine is equally committed to advancing the quality and strength of its medical and graduate education programs, for which it is recognized as an innovative leader, and to training highly skilled, compassionate clinicians and creative scientists well-equipped to engage in world-class research. The School of Medicine is the academic partner of UPMC, which has collaborated with the University to raise the standard of medical excellence in Pittsburgh and to position health care as a driving force behind the region’s economy. For more information about the School of Medicine, see www.medschool.pitt.edu.

 

Researchers discover how we perceive bitter taste



UNIVERSITY OF NORTH CAROLINA HEALTH CARE
CryoEm of TTAS2R14-Ggust-scFv16 

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REPRESENTATIVE CRYO-EM MAPS (LEFT) AND MODELS (RIGHT) FOR TAS2R14-GGUST-SCFV16, COLORED ACCORDING TO
EACH SUBUNIT, TAS2R14; GREEN, GΑI1; PURPLE, GΑGUST; PINK, GΒ1; SKY BLUE, GΓ2;YELLOW, SCFV16; GRAY.

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CREDIT: KIM ET AL (2024)





CHAPEL HILL, N.C. – Humans can sense five different tastes: sour, sweet, umami, bitter, and salty, using specialized sensors on our tongues called taste receptors. Other than allowing us to enjoy delicious foods, the sensation of taste allows us to determine the chemical makeup of food and prevents us from consuming toxic substances.

Researchers at the UNC School of Medicine, including Bryan Roth, MD, PhD, the Michael Hooker Distinguished Professor of Pharmacology, and Yoojoong Kim, PhD, a postdoctoral researcher in the Roth Lab, recently set out to address one very basic question: "How exactly do we perceive bitter taste?”

A new study, published in Nature, reveals the detailed protein structure of the TAS2R14 bitter taste receptor. In addition to solving the structure of this taste receptor, the researchers were also able to determine where bitter-tasting substances bind to TAS2R14 and how they activate them, allowing us to taste bitter substances.

“Scientists know very little about the structural make up of sweet, bitter, and umami taste receptors,” said Kim. “Using a combination of biochemical and computational methods, we now know the structure of the bitter taste receptor TAS2R14 and the mechanisms that initializes the sensation of bitter taste in our tongues.”

This detailed information is important for discovering and designing drug candidates that can directly regulate taste receptors, with the potential to treat metabolic diseases such as obesity and diabetes.

From Chemicals to Electricity to Sensation

TAS2R14s are members of the G protein-coupled receptor (GPCR) family of bitter taste receptors. The receptors are attached to a protein known as a G protein. TAS2R14 stands out from the others in its family because it can identify more than 100 distinct substances known as bitter tastants.

Researchers found that when bitter tastants come into contact with TAS2R14 receptors, the chemicals wedge themselves into to a specific spot on the receptor called an allosteric site, this causes the protein to change its shape, activating the attached G protein.

This triggers a series of biochemical reactions within the taste receptor cell, leading to activation of the receptor, which can then send signals to tiny nerve fibers – through the cranial nerves in the face – to an area of the brain called the gustatory cortex. It is here where the brain processes and perceives the signals as bitterness. And of course, this complex signaling system occurs almost instantaneously.

Cholesterol’s Role in Bitter Taste Reception

While working to define its structure, researchers found another unique feature of TAS2R14 – that cholesterol is giving it a helping hand in its activation.

“Cholesterol was residing in another binding site called the orthosteric pocket in TAS2R14, while the bitter tastant binds to the allosteric site,” said Kim. “Through molecular dynamics simulations, we also found that the cholesterol puts the receptor in a semi-active state, so it can be easily activated by the bitter tastant.”

Bile acids, which are created in the liver, have similar chemical structures with cholesterol. Previous studies have suggested that bile acids can bind and activate TAS2R14, but little is known about how and where they bind in the receptor.

Using their newfound structure, researchers found that bile acids might be binding to the same orthosteric pocket as cholesterol. While the exact role of bile acid or cholesterol in TAS2R14 remains unknown, it may play a role in the metabolism of these substances or in relation to metabolic disorders such as obesity or diabetes.

How This Can Help Drug Development

The discovery of this novel allosteric binding site for bitter tasting substances is unique.

The allosteric binding region is located between TAS2R14 and its coupled G protein is called G-protein alpha. This region is critical to form a signaling complex, which helps to transfer the signal from the taste receptor to the G-protein to the taste receptor cells.

“In the future, this structure will be key to discovering and designing drug candidates that can directly regulate G proteins through the allosteric sites,” said Kim. “We also have the ability to affect specific G-protein subtypes, like G-protein alpha or G-protein beta, rather than other G-protein pathways that we don't want to cause any other side effects.”

Roth and Kim have made a number of new discoveries, but some leave more questions than answers. While running a genomics study, they found that the TAS2R14 protein in complex with the GI is expressed outside the tongue, especially in the cerebellum in the brain, the thyroid, and the pancreas. Researchers are planning future studies to elucidate the function these proteins may have outside of the mouth.

This work was supported by the NIH Illuminating the Druggable Genome Initiative.

About UNC School of Medicine

The UNC School of Medicine (SOM) is the state’s largest medical school, graduating more than 180 new physicians each year. It is consistently ranked among the top medical schools in the US, including 5th overall for primary care by US News & World Report, and 6th for research among public universities. More than half of the school’s 1,700 faculty members served as principal investigators on active research awards in 2021. Two UNC SOM faculty members have earned Nobel Prize awards.

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WHITE SUPREMACY KILLS

‘Deaths of despair’ among Black Americans surpassed those of white Americans in 2022



UCLA Health analysis also found that Native Americans had highest combined death rate from suicide, alcohol use and drug overdose from 1999 through 2022



UNIVERSITY OF CALIFORNIA - LOS ANGELES HEALTH SCIENCES




A new analysis by researchers at UCLA Health found that mortality rates of middle-aged Black Americans caused by the “deaths of despair” -- suicide, drug overdose and alcoholic liver disease – surpassed the rate of white Americans in 2022. Native Americans also had more than double the rate of both Black and white Americans that year.

The results of the research, published in the journal JAMA Psychiatry, are the latest from UCLA Health to counter a nearly decade-old narrative that “deaths of despair” have primarily impacted white Americans.

The “deaths of despair” theory rose to prominence following a 2015 study that analyzed rising mid-life mortality, and decreasing life expectancy, in the U.S from 1999 to 2013.

The study became a focus of ongoing national discourse after it found white Americans had the highest mortality rates from these causes at 72.15 per 100,000 people in 2013, which was two times that of Black Americans. The findings sparked a narrative that the rising death rates were primarily impacting less educated white Americans who were experiencing a perceived loss of economic and social status.

However, data for Native Americans were not included in the 2015 study or in the many follow-up analyses it triggered. A 2023 study by UCLA Health determined that Native Americans have had a considerably higher midlife death rate from deaths of despair compared to white Americans for all available years of data 1999-2021.

In the new data analysis, UCLA Health researchers used publicly available records from the U.S. Centers for disease control to determine rates of deaths of despair through 2022 for white, Black and Native Americans ages 45-54.

The analysis found:

  • The rate of deaths of despair among Black Americans (103.81 per 100,000 people) surpassed that of white Americans (102.63 per 100,000) in 2022.
  • The rate among Black Americans tripled from 2013-2022 from 36.24 per 100,000 to 103.81 per 100,000 people, with 2015 onward marking a sharp increase in deaths.
  • The rate for Native American and Alaska Native populations was the highest at 241.7 per 100,000 people in 2022.
  • The rate of deaths of despair for Native American/Alaska Native populations has remained significantly higher than white Americans from 1999 to 2022.
  • Native American/Alaska Native had the highest mortality rates from suicide, alcoholic liver disease and drug overdose compared to Black and white Americans.
  • Deaths of despair in white Americans increased from 72.15 per 100,000 people in 2013 to 102.63 per 100,000 people in 2022.

Study coauthor Joseph Friedman, PhD, MPH, of the David Geffen School of Medicine at UCLA, said the results highlight the sharply rising rates of premature mortality related to mental health issues and substance use disorders with large racial and ethnic inequalities.

“The findings reinforce the notion that we need to invest in services that can address these issues and, ultimately, we need much more comprehensive access to low-barrier mental health care and substance use treatment in the U.S.,” Friedman said. “And we need to specifically make sure those treatments, services and programs are implemented in a way that is accessible for communities of color and will actively work to address inequality.”

The data does not identify the circumstances behind these deaths, but Friedman said the increasing rates of deaths of despair largely relate to mental health and substance use disorders. Black Americans have been disproportionately affected by the drug overdose crisis. Other factors include differential access to health care and social services; an increasingly toxic illicit drug supply, namely fentanyl; and worsening economic insecurity.

Article: Trends in Deaths of Despair by Race and Ethnicity From 1999 to 2022. Published Online: April 10, 2024. doi:10.1001/jamapsychiatry.2024.0303


JAMA NETWORK

Trends in deaths of despair by race and ethnicity



JAMA Psychiatry





About The Study: As of 2022, the midlife mortality rates from deaths of despair (deaths from suicide, drug overdose, and alcoholic liver disease) among Black individuals were higher than rates among white individuals, and rates among American Indian or Alaska Native individuals remained higher than rates in the other groups. Rising inequalities in deaths of despair among American Indian or Alaska Native and Black individuals were largely attributable to disproportionate early mortality from drug- and alcohol-related causes, which increased leading up to and during the COVID-19 pandemic. 

Authors: Joseph Friedman, Ph.D., M.P.H., of the University of California, Los Angeles, is the corresponding author.

To access the embargoed study: Visit our For The Media website at this link https://media.jamanetwork.com/

(10.1001/jamapsychiatry.2024.0303)

Editor’s Note: Please see the article for additional information, including other authors, author contributions and affiliations, conflict of interest and financial disclosures, and funding and support.

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Embed this link to provide your readers free access to the full-text article This link will be live at the embargo time 

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Race, ethnicity, and delayed time to COVID-19 testing among health care workers


JAMA NETWORK





About The Study: In this cross-sectional study of health care personnel (HCP), compared with non-Hispanic white HCP and clinical HCP with graduate degrees, non-Hispanic Black HCP, non-Hispanic HCP of other races, and HCP of all other professional and education backgrounds were more likely to have delayed COVID-19 testing. These findings suggest that time to testing may serve as a valuable metric in evaluating sociodemographic disparities in the response to COVID-19 and future health mitigation strategies. 

Authors: DaMarcus E. Baymon, M.D., of Brigham and Women’s Hospital in Boston, is the corresponding author. 

To access the embargoed study: Visit our For The Media website at this link https://media.jamanetwork.com/ 

(doi:10.1001/jamanetworkopen.2024.5697)

Editor’s Note: Please see the article for additional information, including other authors, author contributions and affiliations, conflict of interest and financial disclosures, and funding and support.

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Embed this link to provide your readers free access to the full-text article This link will be live at the embargo time


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About JAMA Network Open: JAMA Network Open is an online-only open access general medical journal from the JAMA Network. On weekdays, the journal publishes peer-reviewed clinical research and commentary in more than 40 medical and health subject areas. Every article is free online from the day of publication. 

 

Living near green space associated with fewer emotional problems in preschool-age kids, NIH study finds



ENVIRONMENTAL INFLUENCES ON CHILD HEALTH OUTCOMES
ECHO Program 

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NIH ECHO PROGRAM 

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CREDIT: NIH ECHO PROGRAM






Children who live in areas with natural spaces (e.g., forests, parks, backyards) from birth may experience fewer emotional issues between the ages of 2 and 5, according to a study funded by the NIH Environmental Influences on Child Health Outcomes (ECHO) program.

While research has suggested that time in nature is important for mental health, studies examining the effects on young children are limited. ECHO investigators addressed this research gap by analyzing information from parents about the behavior of their children from ages 2 to 11. They combined this data with the family’s residential address when the child was born and satellite data on live vegetation density around their homes.

What researchers found in their analysis, published in JAMA Network Open, was that higher levels of green spaces up to three-fourths of a mile from a child’s home were linked with lower anxiety and depression symptoms from ages 2 to 5 years. The association persisted even after researchers factored in the child’s sex, parent education, age at birth, and neighborhood socioeconomic vulnerability. Researchers did not find a significant association between green space around the home and mental health symptoms in later childhood years from ages 6 to 11, when children spend more time at school.

“Our research supports existing evidence that being in nature is good for kids,” said Nissa Towe-Goodman, PhD, an ECHO researcher from the Frank Porter Graham Child Development Institute at the University of North Carolina, Chapel Hill. “It also suggests that the early childhood years are a crucial time for exposure to green spaces.”

Most research so far has been limited to studying one or a few cities at a time, and focused on adult health. Because the ECHO Program collects data nationwide, researchers were able to examine data from children in 199 counties across 41 U.S. states, exploring the connection between exposure to green spaces from birth and anxiety, depression, aggression, and other symptoms during early or middle childhood.

The study included children born between 2007 and 2013 and whose parents completed the Child Behavior Checklist, a common survey to rate a child’s emotional and behavioral symptoms. The 2,103 children included in the study ranged in age from 2 to 11, spanning early and middle childhood.

Green space exposure was measured using the Normalized Difference Vegetation Index (NDVI), a widely used metric for quantifying vegetation density using sensor data. NDVI values range from -1 to 1. High NDVI values (approximately 0.6 to 0.9) represent dense vegetation, such as forests; values close to zero represent areas without live vegetation.

 

“In the future, researchers could look into what kinds of experiences in nature are connected to kids' early mental health,” said Dr. Towe-Goodman. “Also, we should study how creating or preserving natural areas around homes and schools might make a difference in a child’s mental health.”

Dr. Towe-Goodman lead this collaborative research in JAMA Network Open.

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About ECHO:

Launched in 2016, the Environmental influences on Child Health Outcomes (ECHO) Program is a research program in the Office of the Director at the NIH with the mission to enhance the health of children for generations to come. ECHO investigators study the effects of a broad range of early environmental influences on child health and development. For more information, visit echochildren.org.

About the NIH: NIH, the nation’s medical research agency, includes 27 Institutes and Centers and is a component of the U.S. Department of Health and Human Services. NIH is the primary federal agency conducting and supporting basic, clinical, and translational medical research, and is investigating the causes, treatments, and cures for both common and rare diseases. For more information, visit www.nih.gov.