Researchers identified two miRNAs that are downregulated in the disease and an uncommon situation that occurs in which multiple miRNAs regulate the same set of genes. CREDIT Reiko Matsushita

A group of researchers from the Graduate School of Medicine at Nagoya University in Japan have discovered the impact of microRNA (miRNA) on inflammation in lupus in mice. They identified two miRNAs that are downregulated in the disease and an uncommon situation that occurs in which multiple miRNAs regulate the same set of genes.  

Although the human body has many types of RNA, the most important is messenger RNA, which is involved in the creation of proteins in the body. The body also contains miRNA, which binds to regions of the messenger RNA to inhibit protein production and regulate several important bodily functions such as development, growth, and metabolism. Problems with miRNA are associated with several diseases including cancer and HIV. Now, the Nagoya University research group has identified the role of miRNA in systemic lupus erythematosus, a disease in which the human immune system attacks itself. They published their findings in BMC Biology.  

Pairing with the correct messenger RNA target is determined by the ‘seed’ of miRNA, a sequence that determines whether the miRNA can bind or not. The seed is like a ‘key’ to the messenger RNA’s ‘lock.’ However, this is complicated by the nature of miRNA’s interaction with messenger RNA as a single particle of messenger RNA may be regulated by multiple miRNAs and the miRNA-messenger RNA pairs do not have to be an exact match to exert an effect. 

As the effects of a single miRNA on a binding receptor site tend to be modest, stronger effects are often regulated by multiple miRNAs working in concert. This occurs through two processes. The first of these processes is ‘neighborhood’ miRNA co-targeting, where two nearby miRNAs affect messenger RNA. The second, is ‘seed overlap’ miRNA co-targeting, which is similar to the neighborhood type except both have similar nucleotides, so they bind to messenger RNA in such a way that some of their nucleotides overlap.  

Given that altered miRNA expression has been reported in lupus disease, researchers have long suspected a connection. Now, a group of scientists, headed by Professor Hiroshi Suzuki at the Department of Molecular Oncology, and Lecturer Noritoshi Kato and Researcher Hiroki Kitai at the Department of Nephrology at the Nagoya University Graduate School of Medicine, have performed miRNA expression profiling using mice with lupus to investigate the role of miRNA in the disease.  

The researchers found that two microRNAs, miR-128 and miR-148a, were down-regulated in plasmacytoid dendritic cells in lupus patients. As plasmacytoid dendritic cells play a crucial role in antiviral immunity and antibody production, they have been implicated in the initiation and development of several autoimmune and inflammatory diseases, including lupus. Both miR-128 and miR-148a target a gene called KLF4, which is associated with inflammatory control and the production of cytokines that regulate the activity of the immune system. 

“Assuming that the expression levels of the other miRNA are maintained, the downregulation of one miRNA can be compensated for by the other microRNA,” Suzuki explains. “However, when two miRNAs decrease simultaneously, as in lupus disease, alterations in their target — in this case KLF4 — emerge.” 

One of the most important findings of the study was that as miR-128 and miR-148a share common nucleotides, they can bind to messenger RNA using ‘seed overlap’ miRNA co-targeting.  “miR-128 and miR-148a target KLF4 through extensive ‘seed overlap’ miRNA cotargeting. In this case, it negatively regulates the production of inflammatory cytokines,” says Suzuki. “Therefore, this study collectively suggests the complexity of different modes of miRNA cotargeting and the importance of their perturbations in human diseases.” 

The researchers also performed integrative analyses, discovering that “seed overlap” miRNA cotargeting of KLF4 is a prevalent feature in other species. “We found that the conserved overlap site of KLF4 is the same in most species between humans and Coelacanths,” said Suzuki. “Therefore, we expanded these findings by integratively analyzing seed overlap patterns of all miRNAs and the conservation patterns of ‘seed overlap’ target sites.” 

Suzuki and the research team discovered two main conservation classes of miRNA target sites. The first was shared by eutherian mammals, including animals that have a placenta. The second was shared by other animals, including humans and Coelacanths, and has a stronger association with both “seed overlap” and “neighborhood” miRNA cotargeting.  

“Our study provides a comprehensive view of ‘seed overlap’ miRNA cotargeting, which is very important for the process by which lupus develops from the viewpoint of gene regulation and miRNA evolution. These findings highlight the importance of miRNA co-targeting in human pathology and the unique evolutionary aspects of miRNA co-targeting and miRNA target site conservation,” Suzuki explains.  

Suzuki also sees the potential of his research in treating lupus patients: “Testing for downregulation of the two miRNAs may help identify patients with high level of inflammation who may benefit from specific therapeutic development,” he says.  

The frequency and size of meals were a more vital determinant of weight loss or gain than the time between the first and last meal, according to new research published today in the Journal of the American Heart Association, an open-access, peer-reviewed journal of the American Heart Association.

According to the senior study author Wendy L. Bennett, M.D., M.P.H., an associate professor of medicine at Johns Hopkins University School of Medicine in Baltimore, although ‘time-restricted eating patterns’ – known as intermittent fasting – are popular, rigorously designed studies have not yet determined whether limiting the total eating window during the day helps to control weight.

This study evaluated the association between time from the first meal to the last meal with weight change. Nearly 550 adults (18 years old or older) from three health systems in Maryland and Pennsylvania with electronic health records were enrolled in the study. Participants had at least one weight and height measurement registered in the two years prior to the study’s enrollment period (Feb.-July 2019).

Most participants (80%) reported they were white adults; 12% self-reported as Black adults; and about 3% self-identified as Asian adults. Most participants reported having a college education or higher; the average age was 51 years, and the average body mass index was 30.8, considered obese. The average follow-up time for weight recorded in the electronic health record was 6.3 years.

Participants with a higher body mass index at enrollment were more likely to be Black adults, older, have Type 2 diabetes or high blood pressure, have a lower education level, exercise less, eat fewer fruits and vegetables, have a longer duration from last mealtime to sleep and a shorter duration from first to last meal, compared to the adults who had a lower body mass index.

The research team created a mobile application, Daily24, for participants to catalogue sleeping, eating and wake-up time for each 24-hour window in real-time. Emails, text messages and in-app notifications encouraged participants to use the app as much as possible during the first month and again during “power weeks” — one week per month for the six-month intervention portion of the study.

Based on the timing of sleeping and eating each day recorded in the mobile app, researchers were able to measure:

• the time from the first meal to the last meal each day;
• the time lapse from waking to first meal; and
• the interval from the last meal to sleep.

They calculated an average for all data from completed days for each participant.

The data analysis found:

• Meal timing was not associated with weight change during the six-year follow-up period. This includes the interval from first to last meal, from waking up to eating a first meal, from eating the last meal to going to sleep and total sleep duration.
• Total daily number of large meals (estimated at more than 1,000 calories) and medium meals (estimated at 500-1,000 calories) were each associated with increased weight over the six-year follow up, while fewer small meals (estimated at less than 500 calories) was associated with decreasing weight.
• The average time from first to last meal was 11.5 hours; average time from wake up to first meal measured 1.6 hours; average time from last meal to sleep was 4 hours; and average sleep duration was calculated at 7.5 hours.
• The study did not detect an association meal timing and weight change in a population with a wide range of body weight.

As reported by Bennett, even though prior studies have suggested intermittent fasting may improve the body’s rhythms and regulate metabolism, this study in a large group with a wide range of body weights did not detect this link. Large-scale, rigorous clinical trials of intermittent fasting on long-term weight change are extremely difficult to conduct; however, even short-term intervention studies may be valuable to help guide future recommendations.

Although the study found that meal frequency and total calorie intake were stronger risk factors for weight change than meal timing, the findings could not prove direct cause and effect, according to lead study author Di Zhao, Ph.D., an associate scientist in the division of cardiovascular and clinical epidemiology at Johns Hopkins Bloomberg School of Public Health.

Researchers note there are limitations to the study since they did not evaluate the complex interactions of timing and frequency of eating. Additionally, since the study is observational in nature, the authors were unable to conclude cause and effect. Future studies should work toward including a more diverse population, since the majority of the study’s participants were well-educated white women in the mid-Atlantic region of the U.S., the authors noted author.

Researchers also were not able to determine the intentionality of weight loss among study participants prior to their enrollment and could not rule out the additional variable of any preexisting health conditions.

According to the American Heart Association’s 2022 statistics, 40% of adults in the U.S. are obese; and the Association’s current diet and lifestyle recommendations to reduce cardiovascular disease risk include limiting overall calorie intake, eating healthy foods and increasing physical activity.

New research suggests coaching overweight or obese pregnant women to improve their ability to plan and progress toward goals may be vital to helping them lower the amount of fat in their diet.

Maternal diet quality affects prenatal development and long-term child health outcomes. Still, the stress that typically increases during pregnancy – often heightened by concern for fetal health and anxiety over impending parenthood – may derail efforts to focus on healthful eating, previous research has shown.  

In this new study, researchers at The Ohio State University set out to identify the pathway between stress and total fat consumption, with a broader goal to evaluate an intervention designed to improve the diets of pregnant women who are overweight or obese.

Through a series of questionnaires and statistical analysis, the team found that two thinking-related skills – planning and execution of those plans – were weakened in women whose stress was high, and those skill gaps were associated with higher total fat intake.

These two skills are executive functions, a set of multiple thinking processes that enable people to plan, monitor behavior and execute their goals.

“People with a higher stress level tend to have a higher fat intake, too. If stress is high, we’re so stressed out that we’re not thinking about anything – and we don’t care what we eat,” said lead author Mei-Wei Chang, associate professor of nursing at Ohio State.

“That’s why we focused on executive functions as a mediator between stress and diet. And with this baseline data, we have reasons to believe that designing an intervention around executive functions could improve dietary outcomes,” she said. “I would anticipate the results could be similar for nonpregnant women because it’s all about how people behave.”

The study was published recently in the Journal of Pediatrics, Perinatology and Child Health.

The 70 women enrolled in the study had a pre-pregnancy body mass index of between 25 (scores between 25 and 29.9 are categorized as overweight) and 45 (scores of 30 and higher are categorized as obese).

The participants completed questionnaires assessing both overall perceived stress and pregnancy-related stress, as well as executive functions – specifically focusing on metacognition, the ability to plan, and behaviour regulation, the ability to execute those plans. They also completed two 24-hour dietary recalls of their calorie intake and consumption of total fat, added sugar, and fruits and vegetables.

“We were interested in the mediation role of executive functions. The mediator is what makes everything happen,” Chang said. “We wanted to know: If we focus intervention on executive functions, would that carry through to behaviour change in dietary intake?

“Weight loss interventions often involve a prescribed diet or meal plan, and you are told to follow it. But that doesn’t lead to behaviour change in the long term.”

Statistical modelling showed that higher perceived stress was associated with a worsened ability to plan and monitor behaviour, and that pathway was linked to higher total fat intake. Similarly, higher levels of pregnancy-related stress were associated with a lower ability to plan, which in turn was associated with the worsened ability to monitor behaviours related to carrying out the plan – and these factors were linked to higher fat consumption.

These pathways suggested that an intervention designed to lower stress would function as a starting point to improve the diet and enhancing skills through coaching – emphasizing the ability to plan, including being flexible with planning, and behaviour monitoring, particularly when making food choices – would be key to changing eating patterns.

“You need to improve executive functions, and you also need to lower stress,” Chang said. She and colleagues are now analyzing data on the effectiveness of an intervention for the study participants that emphasized stress management and boosting executive function to promote healthy eating.

A specific region of the brain regulates executive functions, and strengths or weaknesses in these skill areas are thought to be affected by various physiological factors. Previous research has found that executive function deficits are more likely to occur in women who are overweight or obese than in women whose weight is average.

“Executive function is not well-studied, and it is not related to intelligence. But people with low executive function cannot make detailed plans and stick to them, and that’s how they get into trouble,” Chang said. “Metacognition and behaviour regulation must go hand in hand – that way you have a much better chance to control your behaviours, and then you will eat better.”

A first study of its kind by Tel Aviv University researchers examined the “secret magic” of medical clowns

According to researchers from Tel Aviv University and the Israel Center for Medical Simulation: “medical clowns not only help the patients, but they also help the parents, the medical team, and the achievement of therapeutic goals. In fact, through various communication skills, clowns enable patients to overcome crises and move towards healing.”

Not just fun and games: a new study tested and categorized the skills of medical clowns found that their importance goes far beyond just creating a good mood for patients. The researchers identified 40 different skills of medical clowns, including establishing an emotional connection and creating a personal relationship with the patient, expressing the patient’s voice about his/her frustrations and difficulties to the medical staff, increasing the patient’s motivation to adhere to the medical treatment, distracting the patient from pain, and creating an atmosphere of games and play. The findings emphasize that the importance of clowns goes far beyond “providing just a good mood” for patients.

The research was conducted under the leadership of Prof. Orit Karnieli-Miller, with Dr. Lior Rosenthal, both from the Department of Medical Education at Tel Aviv University, in collaboration with Ms. Orna Divon-Ophir, Dr. Doron Sagi, Prof. Amitai Ziv and Ms. Liat Pessach-Gelblum from the Israel Center for Medical Simulation (MSR). The study was published in Qualitative Health Research, a leading journal in the field of health.

Prof. Orit Karnieli-Miller CREDITTel Aviv University

The researchers show that medical clowns not only help the patients and their family members, but also the medical team and the achievement of treatment goals. Through use of different communication skills, clowns make it easier for the patient to cooperate with one or another treatment. The medical clowns work in a team with other therapists and know how to intervene and help when an argument or crisis arises, and help overcome it and advance treatment.

Earlier studies showed medical clowns as trained professionals whose goal is to change the hospital environment through humor. Studies conducted throughout the years have shown the clowns’ positive influence on this and in helping patients deal with pain. However, so far, no studies have empirically mapped the skills they use and their therapeutic goals to help understand their “secret magic.” In addition, there was a lack of broad understanding of how clowns can help children, teenagers, and their parents in various challenging situations of distress and difficulty, as well as how they can help patients and medical teams achieve treatment goals. This lack of appreciation of the potential benefits of utilizing the services of a medical clown meant that sometimes patients and medical teams could be reluctant to cooperate with them.

As part of the new study, the team of researchers focused on qualitative, in-depth systematic identification of the skills of medical clowns through observation and analysis of their actions in challenging encounters with adolescents, parents, and medical staff. The team analyzed videotaped sessions of medical clowns in various simulated situations and conducted in-depth interviews with expert medical clowns. The researchers identified forty different skills used by the medical clowns to achieve four therapeutic goals: 1) building a relationship and connecting to the needs and desires of the patients; 2) dealing with emotions and difficulties; 3) increasing the patient’s motivation to adhere to the treatment plan; and 4) increasing the patient’s sense of control and providing encouragement to patients.

The clowns examined in the study were trained and recruited by the “Dream Doctors Project”, a non-profit association that employs medical clowns as part of the paramedical system in Israeli hospitals, and trains them to work within multi-disciplinary teams. The Tel Aviv University researchers collaborated with the Israel Center for Medical Simulation (MSR), which created a simulation-based workshop focused on developing the skills of experienced medical clowns.

Prof. Karnieli-Miller: “from the moment they enter the room, the clowns form a bond with the patients, strengthen them, and give them power and status within the medical system. They do this through an initial connection to the patients’ voice, and even to the patients’ reluctance to implement therapeutic recommendations – an emotional connection that often results in the patient changing their position and cooperating with the medical staff. The medical system is hierarchical, and it is not always easy for patients to navigate. Therefore, one of the skills of medical clowns is to place themselves in the lowest position in the medical setting. By doing so they empower the patients, giving the patients power and control, including the choice of whether to allow the clown to enter the room as well as to dictate the nature of the patient’s role vis-à-vis that of the clown. This provides an increased sense of control and gives the patient courage to face their challenges.”

The researchers emphasize that the clowns are very aware of the emotional difficulty associated with staying in a hospital and dealing with an illness. To deal with these issues the clowns sometimes distract the patient by using props, humor, and imagination. Other skills include the clowns allowing the patient to direct their frustrations towards them, away from the medical staff or parents. Depending on the situation the clowns may also use a comforting touch, soothing music, empathetic listening, or a reinforcing statement to provide an environment where the patient feels comfortable to express their feelings. A patient’s ability to gain legitimacy is important and is strengthened by the clowns. Prof. Karnieli-Miller adds: “Mapping the skills and goals of the medical clowns improves their understanding of their role and may help other health professionals appreciate their work methods and the benefits of incorporating these methods into their own practices when faced with similar challenges.”

Prof. Karnieli-Miller concludes: “This research is important because it allows the clowns to enhance their training program and refine their diverse skills to achieve the various therapeutic goals appropriate for different patients, as well as helping health professionals collaborate with the medical clowns. If professionals in the healthcare field will know how and when to cooperate with the medical clowns, they will be able to help patients overcome challenges, and at the same time they may be more tolerant of the clowns’ ‘disruption’ of the hospital care regimen. This appreciation of the clowns’ contribution will provide the clowns with the time and space to connect with patients and help and encourage patients to become more active participants in their treatment plan. Important goals all health professionals aspire to.”

Chemical “micromotors,” as illustrated here, can effectively deliver insulin in rats without an injection. CREDIT Adapted from ACS Nano, 2022, DOI: 10.1021/acsnano.2c07953

For millions of people living with diabetes, insulin is a life-saving drug. Unlike many other medicines, though, insulin cannot be easily delivered by swallowing a pill — it needs to be injected under the skin with a syringe or pump. Researchers have been making steps toward an insulin pill. A team reports in ACS Nano that they’ve delivered insulin to the colons of rats using an orally administered tablet powered by chemical “micromotors.”

Patients with diabetes have trouble regulating their blood glucose levels because they produce little or no insulin. Synthetic insulin has existed for over a hundred years, but it is often administered with an injection or an implanted pump. People affected by diabetes often take insulin multiple times per day so frequent injections can be painful, and as a result, some patients do not take the recommended dose at the correct times.

An oral form of the drug would be ideal, but the stomach’s harsh environment breaks down and neutralizes the hormone before it can be absorbed by the intestines and get into the bloodstream. Previous attempts at oral administration protected the hormone from stomach acids with micro- or nanocarriers. Still, they relied on insulin to diffuse passively into the cells that line the colon, which isn’t very efficient. A better approach could be actively moving the medicine around the body instead, such as with a recently reported Robo-capsule that delivers its cargo by drilling itself into the thick mucosal layer of the small intestine. Yingfeng Tu, Fei Peng, Kun Liu and colleagues wanted to achieve a similar effect with their insulin-loaded mini-tablets, which featured tiny, chemical “micromotors” that could deliver insulin to the colon safely and effectively.

To make these tablets, the researchers covered magnesium microparticles with a layer of an insulin-containing solution and a layer of liposomes. They then mixed these particles with baking soda, pressed them into mini-tablets about 3 mm long then covered them with an esterified starch solution. The starch protected the tablets from stomach acid, allowing them to reach the colon intact. As they broke down, the magnesium microparticles reacted with water to generate a stream of hydrogen gas bubbles, which acted as micromotors that propelled insulin toward the colon’s lining to be absorbed. The team also tested their mini-tablets in rats and found that they could significantly reduce the animals’ blood glucose levels for over five hours. In fact, they could maintain a glucose level almost as low as injection-delivered insulin. Though more work is needed, the researchers say that this is a concrete step toward creating more oral formulations of traditionally injection-only medications.