Fluorescent Detection of Reactive Oxygen Species in \u3cem\u3eSaccharomyces cerevisiae\u3c/em\u3e Applied to Chronological Lifespan

During the course of normal aerobic metabolism, cells are exposed to a wide range of reactive oxygen species such as the superoxide anion, hydrogen peroxide, and the hydroxyl radical. These reactive oxygen species (ROS) are highly reactive metabolites of oxygen and can damage a wide range of macromolecules in the cell, including nucleic acids, proteins, and lipids, and can even, in some severe cases, lead to cell death. Normally, molecular oxygen is relatively unreactive and harmless in its ground state; however, it can undergo partial reduction via electrons that are leaked from the electron transport chain to form both the superoxide anion and hydrogen peroxide, both of which can react further to form the dangerously reactive hydroxyl radical. In order to combat the toxic and potentially deadly effects of ROS, cells are equipped with various antioxidant defense mechanisms, which include enzymes like superoxide dismutase 1 (∆sod1). Our objective is to observe these various reactive oxygen species using yeast (Saccharomyces cerevisiae) as a model organism and explore different biochemical staining assays such as Amplex Red (AR) and Dihydroethidium (DHE). These stains can both be used to track live cells and quantify ROS levels. This will allow us to study how ROS changes during chronological yeast lifespan. Although there are many types of reactive oxygen species that exist in various parts of the cell, our work thus far has aimed to track extracellular hydrogen peroxide via AR and superoxide generation in the mitochondria via DHE. Our initial results indicate that we are able to track superoxide production using DHE in wild type cell and ∆sod1 yeast strains spectroscopically. Ultimately, we will use both fluorescence spectroscopy and live cell imaging via fluorescence microscopy to assess superoxide levels in multiple yeast strains. Our results will provide insight into the role of ROS in aging as we quantify levels during yeast lifespan

Fluorescent Detection of Reactive Oxygen Species in Saccharomyces Cerevisiae Applied to Chronological Lifespan

During the course of normal aerobic metabolism, cells are exposed to a wide range of reactive oxygen species such as the superoxide anion, hydrogen peroxide, and the hydroxyl radical. These reactive oxygen species (ROS) are highly reactive metabolites of oxygen and can damage a wide range of macromolecules in the cell, including nucleic acids, proteins, and lipids, and can even, in some severe cases, lead to cell death. Normally, molecular oxygen is relatively unreactive and harmless in its ground state; however, it can undergo partial reduction via electrons that are leaked from the electron transport chain to form both the superoxide anion and hydrogen peroxide, both of which can react further to form the dangerously reactive hydroxyl radical. In order to combat the toxic and potentially deadly effects of ROS, cells are equipped with various antioxidant defense mechanisms, which include enzymes like superoxide dismutase 1 (Sod1p). Our objective is to observe these various reactive oxygen species using yeast (Saccharomyces cerevisiae) as a model organism and explore different biochemical staining assays such as Amplex Red (AR) and Dihydroethidium (DHE). These stains can both be used to track live cells and quantify ROS levels. This will allow us to study how ROS changes during chronological yeast lifespan. Although there are many types of reactive oxygen species that exist in various parts of the cell, our work thus far has aimed to track extracellular hydrogen peroxide via AR and superoxide generation in the mitochondria via DHE. Our initial results indicate that we are able to track superoxide production using DHE in wild type cell and sod1∆ yeast strains spectroscopically. Ultimately, we will use both fluorescence spectroscopy and live cell imaging via fluorescence microscopy to assess superoxide levels in multiple yeast strains. Our results will provide insight into the role of ROS in aging as we quantify levels during yeast lifespan

Thriving Private-Community Partnerships: Perspectives from Fiji’s Upper Navua Conservation Area

Ecotourism has proven itself to be a forerunner in the advancement of environmental conservation all the while supporting cultural tradition, uniqueness, and pride among indigenous communities. Successful private-community partnerships associated with ecotourism operations are vital to the overall prosperity of both the businesses and the local communities. Such accomplishments can be seen through numerous livelihood goals including income, food security, health, reduced vulnerability, governance, and empowerment (Walpole & Wilder, 2008). Private-community partnerships also support global initiatives such as the Sustainable Development Goals (Ban Ki-Moon, 2015) and sustainable development frameworks including those proposed by the United Nations World Tourism Organization (UNWTO, 2015). Understanding such partnerships assists not only large organizations such as the WTO, but it also benefits smaller ecotourism operators and entrepreneurs who are trying to achieve their sustainable tourism development goals. This study examined the partnership between an ecotourism company (Rivers Fiji) and two rural villages located in Fiji’s Upper Navua Conservation Area. Focus groups were conducted in each village. Observation journals were also used to record conversations outside of the focus groups. Data were thematically organized and analyzed to offer researcher interpretations and understandings

Joint Action Effects of Emamectin Benzoate and Cypermethrin on the Marine Copepod Tigriopus californicus

This project examined the effect of two pesticides on the marine copepod Tigriopus californicus. The pesticides are emamectin benzoate and cypermethrin, which are commonly used in salmon aquaculture to treat the salmon for sea lice, a type of parasitic copepod. The results of this study indicate that these pesticides are toxic to the free-living T. californicus at very low dosage levels when used individually. When the pesticides are used in combination, the mortality rate is similar to when they are used individually, but there is an additional paralytic effect on the animals. All animals exposed to any of the pesticide mixtures used in this study were irreversibly paralyzed and therefore effectively killed. This indicates a strong need for more testing on the combined impacts of pesticides and increased regulatory action for salmon aquaculture

Scaling prediction errors to reward variability benefits error-driven learning in humans.

Effective error-driven learning requires individuals to adapt learning to environmental reward variability. The adaptive mechanism may involve decays in learning rate across subsequent trials, as shown previously, and rescaling of reward prediction errors. The present study investigated the influence of prediction error scaling and, in particular, the consequences for learning performance. Participants explicitly predicted reward magnitudes that were drawn from different probability distributions with specific standard deviations. By fitting the data with reinforcement learning models, we found scaling of prediction errors, in addition to the learning rate decay shown previously. Importantly, the prediction error scaling was closely related to learning performance, defined as accuracy in predicting the mean of reward distributions, across individual participants. In addition, participants who scaled prediction errors relative to standard deviation also presented with more similar performance for different standard deviations, indicating that increases in standard deviation did not substantially decrease "adapters'" accuracy in predicting the means of reward distributions. However, exaggerated scaling beyond the standard deviation resulted in impaired performance. Thus efficient adaptation makes learning more robust to changing variability.This work was supported by the Wellcome Trust and the Niels Stensen Foundation.This is the final version of the article. It first appeared from the American Physiological Society via http://dx.doi.org/10.1152/jn.00483.201

Rheological Characterization Of Dynamic Re–Engineering Of The Pericellular Region By Human Mesenchymal Stem Cell–Secreted Enzymes In Well–Defined Hydrogel Scaffolds

Human mesenchymal stem cells (hMSCs) are critical for wound healing. hMSCs are active in each phase of wound healing regulating inflammation. Inflammatory regulation also enables hMSCs to restart stalled healing in chronic wounds. Before hMSCs reach a wound, they migrate out of their niche and across several rheologically distinctive microenvironments. The aim of synthetic implantable wound healing scaffolds is to mimic these in vivo microenvironments to enhance delivery of hMSCs to the wound while providing structure to the surrounding tissue. To use these materials as implantable wound healing scaffolds, we must determine how to leverage the material microenvironment to encourage cell migration and cell delivery to the wound. These materials mimic both physical and chemical aspects of the native extracellular matrix and enable precise engineering of the cues initially presented in the microenvironment to 3D encapsulated cells. Although these scaffolds are initially well-defined, they are designed to allow cellular remodeling and degradation, which constantly presents new environmental cues to the cell. In our work, we encapsulate human mesenchymal stem cells (hMSCs) into a synthetic poly(ethylene glycol (PEG)-peptide hydrogel scaffold. This scaffold consists of a 4-arm star PEG backbone end-functionalized with norbornene which is cross-linked with a matrix metalloproteinase (MMP) degradable peptide sequence. The encapsulated hMSCs secrete MMPs, enabling them to degrade the hydrogel scaffold prior to and during motility. To characterize cellular re-engineering of the pericellular region we use multiple particle tracking microrheology (MPT) to measure the dynamic changes in the material temporally and spatially. In MPT, 1 micron fluorescently labeled probe particles are embedded in the material and the Brownian motion of these particles is measured and used to determine material properties. Our work measures hMSC-mediated degradation in the pericellular region has a microenvironment where the cross-link density decreases as distance from the cell increases. This is called a reverse reaction-diffusion degradation profile. The hMSC is keeping the scaffold stiff directly around it to spread and attach prior to motility. To do this, the cell simultaneously secretes tissue inhibitors of metalloproteinases (TIMPs), which bind to the catalytic part of the MMP making it inactive and unable to degrade the scaffold. This MMP-TIMP complex then diffuses through the scaffold and unbinds, making the MMP active and degrading the scaffold. This is modeled using Michaelis-Menten competitive inhibition. We then inhibit TIMPs to determine how this changes the rheological properties in the pericellular region and hMSC motility. After TIMP inhibition, we measure a reaction-diffusion degradation profile in the pericellular, which is the opposite of the degradation profile created by untreated hMSCs. Additionally, cell motility is greatly enhanced in the scaffold. This could be due to durotaxis, the migration along a stiffness gradient to a higher moduli material, or due to a decreased material barrier in migration. This simple chemical treatment has the potential to enhance hMSC delivery to wounds from this hydrogel scaffold but it is unclear if the change in the bulk properties of the scaffold could increase degradation to the point where the scaffold can no longer support the wounded tissue if used for implantation

Understanding parents’ sense-making of their role in adolescent daughters’ social media use through the lens of relational dialectics theory 2.0

With the development of social media, parents must figure out how to guide their children’s use or even whether to allow it. Data were gathered through semi-structured interviews with 30 parents of daughters aged 12–18. Relational dialectics theory 2.0 was used to analyze how parents’ talk revealed their sense-making of their role in adolescent daughters’ social media use. Analysis revealed that parents voiced competing discourses pertaining to “bad” versus “good” parents and monitoring versus trusting daughters. Discourses that take place at the distal level compete with those at the proximal site of the utterance chain, challenging parents to engage in sense-making. Findings suggest that the advice of open communication between parent and adolescent addresses only the proximal level and not the distal level of societal expectations for monitoring and close involvement required of the “good parent.” To make sense of and manage competing discourses, parents appear to couple conversations with daughters with voicing discourses of daughter uniqueness as a way to favor trusting over monitoring and still maintain a “good” parent identity

Developmental Boxes

Developmental Boxes The assigned facility is a resource center that operates to assist families in need. Services offered include housing placement, homelessness prevention, securement of affordable childcare, after-school programs, and health education. The center has served the Mount Baker community for nearly three years and intends to continue partnering with public services to end the cycle of poverty and create brighter futures for those who need it most (Mercy Housing, 2021). Our team conducted a windshield survey of the community served by the facility and determined there to be limited access to childcare in the area as well as minimal resources to promote healthy childhood development. With these findings, our team created an intervention aiming to positively affect early childhood development by meeting age-appropriate milestones. To meet this goal, we created developmental boxes for children ages 0-5 with basic care supplies, age-appropriate toys, and educational pamphlets for parents to be better equipped in caring for their young children. Data from the windshield survey showed that 33.3% of residents in the area are unemployed, 33.2% of the population are people of color, and 10% live below the poverty line (Statistical Atlas, n.d.). According to the staff members crime rates have recently been increasing, making it unsafe for children. Families with children make up 27% of the population (Statistical Atlas, n.d.). Numerous families are struggling with finances and are at risk for falling below the poverty line. These statistics imply that parents may struggle to provide appropriate developmental resources for their children. Homelessness and poverty have been proven to adversely affect children’s development as housing is a social determinant of health. Studies have shown that 37% of school-aged children who are from homeless families are falling below average in their schoolwork and 40% of those children have had to repeat grade levels (Murran & Brady, 2023). An association between poverty, children’s development and academic performance is evident as early as two years old. School readiness upon entering kindergarten sets the trajectory for future success (Engle & Black, 2008). Our group assembled developmental boxes targeting children ranging from newborn to 5 years old, including hygiene necessities, clothing, and age-appropriate toys. Due to poverty and homelessness, children in this age group often do not have access to developmental activities to aid in meeting their milestones. According to Erikson’s theory of Development, children in the age group of 0-18 months are expected to have a secure environment provided by their caregiver, in which they can develop in the stage of trust vs. mistrust. Children are expected to develop the skill of trust and hope in their primary caregivers. However, those who experience adverse childhood events such as unstable income, housing, clothing, and food resources are at a greater risk of developing mistrust and having developmental delays. Children in the age group of 18 months to 3 years old are expected to have a caregiver that promotes self-sufficiency while maintaining a secure environment. This helps the child develop a sense of autonomy. However, children from homeless and low-income families may not fully meet this stage due to lack of resources and a secure environment. As a result of lacking a safe home environment to independently explore, clothes to practice dressing themselves, food to begin feeding themselves, toys to independently play with, and access to toilet training, these children may not be equipped with a sense of will, confidence, or autonomy to navigate the world on their own successfully, and may instead experience shame and doubt. Children in the age group of 3 to 5 years old are expected to have a caregiver that encourages, supports, and guides their own initiatives and interests (Orenstein & Lewis, 2023). The purpose of the developmental boxes is to promote proper development for children within this population, who often lack the necessary resources for this development to progress. Specifically, play is a universal childhood activity that fosters development. Play promotes executive function skills by encouraging children to take on cognitive demands, pursue goals, create problems to solve, make choices, and engage in personal interests (Doebel & Lillard, 2023). The developmental boxes that our group assembled include an educational pamphlet focusing on breastfeeding, PURPLE crying, and common childhood illnesses, as well as a map composed of local resources for families. PURPLE crying involves peaks of unexplained, long-lasting crying, which is resistant to soothing (Reese et al., 2014). The population in this area is at a high risk for knowledge deficit due to lack of resources and increased homelessness (Bridget & Oudshoorn, 2019). By providing the pamphlet and resource map in the developmental boxes, parents are receiving education with fundamental information and locations of resources to promote their child’s health and well-being. Common childhood illnesses include the common cold, ear infections, influenza, RSV, and strep throat (Wakeforest Pediatrics Associates, 2022). Additionally, parents may not be aware of specific illnesses their child may have if they are displaying certain symptoms or which symptoms require emergency intervention. The pamphlet also listed symptoms that would require seeking emergency care including coughing up thick, green-yellow phlegm or blood, vomiting, difficulty breathing, severely drowsy, and presenting with blue skin (Raising Children, 2023). Homeless and low-income families often face barriers to accessing health care including transportation, costs, complex systems, communication barriers and not knowing where to go (Bridget & Oudshoorn, 2019). This information is extremely important to supply families with the knowledge they need to support their child’s health. The group set two SMART goals for the project- the first being 75% of families receiving the developmental boxes would report satisfaction with box contents by the end of the quarter. The second project goal was a reduction in screen time from children engaging with the developmental boxes by the end of the quarter. Results from the survey showed 5-10 children engaged with the developmental boxes. Staff and parents reported high satisfaction with the boxes with a reduction in screen time. The facility plans to continue using the developmental boxes following this quarter and rated sustainability to be 4/5. The additional materials provided including the educational pamphlet and resource map were also rated 4/5 as informational and educational. With this data, we conclude that both project goals have been successfully met. The facility exhibited a need for resources related to early childhood development. Our team created an intervention called “Developmental Boxes,” with the aim to decrease screen time in children and provide parents with educational materials to properly care for their young children. Multiple limitations presented themselves during the project- the first being the flooding of the original facility which caused displacement of staff and temporary closure of the site. This prohibited us from serving the originally assigned facility and neighboring community with our intervention. Another limitation that arose was the limited amount of feedback received from staff onsite. This may be due to the displacement of the original staff, and addition of new staff members to the project. Since these staff were introduced late in the quarter, they were unaware of the project idea. Although these limitations made the project challenging, results showed that both project goals were met. The facility reported high ratings for sustainability and plans to continue utilizing the Developmental Boxes once the quarter ends through public donations and federal funding. This project will continue to support future families and young children in their growth and development. References Bridget Osei, H. A., & Oudshoorn, A. (2019). The health challenges of families experiencing homelessness. [Families experiencing homelessness] Housing, Care and Support, 22(2), 93-105. doi:https://doi.org/10.1108/HCS-12-2018-0036 Doebel, S., & Lillard, A.S. (2023). How does play foster development? A new executive function perspective. Developmental Review, 67. https://www.sciencedirect.com/science/article/abs/pii/S0273229722000545?via%3Dihub Engle, P.L., & Black, M.M. (2008). The Effect of Poverty on Child Development and Educational Outcomes (Issue 1, Vol. 1136). Annals of the New York Academy of Sciences. https://doi.org/10.1196/annals.1425.023 Mercy Housing. (2021, July 19). Gardner House and Allen Family Center celebrate one-year anniversary. https://www.mercyhousing.org/2021/06/gardner-house-and-allen-family-center-celebrate-one-year-anniversary/ Murran, S., & Brady, E. (2023). How does family homelessness impact on children’s development? A critical review of the literature. Child & Family Social Work, 28 (2), 360-371. https://doi.org/10.1111/cfs.12968 Orenstein, G.A., & Lewis, L. (2023). Eriksons Stages of Psychosocial Development. StatPearls Publishing. Raising Children. (2023, April 13). Serious childhood illnesses: 0-3 years. Retrieved April 19, 2023, from https://raisingchildren.net.au/babies/health-daily-care/health-concerns/serious-child-illnesses Reese, L.S., Heiden, E.O., Kim, K.Q. & Yang, J. (2014). Evaluation of period of purple crying, an abusive head trauma prevention program. Journal of Obstetric, Gynecologic, & Neonatal Nursing, 43(6), 752-761. https://doi.org/10.1111/1552-6909.12495 Statistical Atlas. (n.d.). Overview of Mt. Baker, Seattle, Washington. Retrieved May 17, 2023, fromhttps://statisticalatlas.com/neighborhood/Washington/Seattle/Mt-Baker/Overview Wakeforest Pediatrics Associates. (2022, April 15). 10 common childhood illnesses. Retrieved April 19, 2023, from https://wakeforestpediatrics.com/common-childhood-illnesses

The common in-group identity model enhances communication about recycled water

Despite the potential of recycled water to provide a safe alternative water source, recycled drinking water schemes have met with community resistance that has undermined their implementation. The aim of the current study was to identify effective means of communicating information about recycled drinking water by drawing on the common in-group identity model. In an online, experimental study, we explored whether awareness of a common/shared superordinate identity could enhance the impact of information on public perceptions of recycled drinking water. The results showed that the communication of information increased participants' acceptance, perceived knowledge and positive emotions toward recycled drinking water and lowered their risk perceptions. Moreover, the results provide the first experimental evidence to show that the effect of information on recycled water was enhanced when the information was attributed to a scientist that shared a superordinate identity with participants, albeit only for those that identify strongly with the superordinate identity