Glucocorticoid response to naturalistic interactions between children and dogs.

Although research has shown that pets appear to provide certain types of social support to children, little is known about the physiological bases of these effects, especially in naturalistic contexts. In this study, we investigated the effect of free-form interactions between children (ages 8-10 years) and dogs on salivary cortisol concentrations in both species. We further investigated the role of the child-dog relationship by comparing interactions with the child's pet dog to interactions with an unfamiliar dog or a nonsocial control condition, and modeled associations between survey measures of the human-animal bond and children's physiological responses. In both children and dogs, salivary cortisol decreased from pre- to post-interaction; the effect was strongest for children interacting with an unfamiliar dog (compared to their pet dog) and for the pet dogs (compared to the unfamiliar dog). We found minimal evidence for associations between cortisol output and behaviors coded from video, but children scoring higher on survey measures of the human-animal bond exhibited the greatest reductions in cortisol when interacting with dogs. Self-reported loneliness was not related to cortisol or the human-animal bond, but measures of both loneliness and the human-animal bond were higher among children who participated after the onset of the COVID-19 pandemic, relative to those who participated before the pandemic. This study builds on previous work that investigated potential stress-buffering effects of human-animal interaction during explicit stressors and demonstrates important physiological correlates of naturalistic interactions between children and dogs, similar to those that occur in daily life.

Diurnal fecal glucocorticoid metabolite rhythms in a cathemeral primate, the red-bellied lemur (Eulemur rubriventer), and across mammalian species.

Measuring glucocorticoids is one of the most reliable and widely used techniques to monitor stress responses, however invasive techniques to collect plasma samples may not be applicable for wild populations. Monitoring excreted glucocorticoids is an effective noninvasive technique that researchers have used increasingly over the past two decades, and it has allowed the investigation of glucocorticoids in a variety of species with a range of activity patterns. Many species exhibit predictable circadian patterns of glucocorticoid secretion in accordance with their daily activity pattern. There remains a gap in our understanding of how excreted glucocorticoid metabolites vary throughout the day and across species, despite the utility of this information when developing sampling protocols and analyzing data. We investigated circadian patterns of glucocorticoid excretion in a cathemeral primate species, Eulemur rubriventer (red-bellied lemur), in Ranomafana National Park, Madagascar. We collected fecal samples from 10 individuals throughout the day and analyzed fecal glucocorticoid levels across three time points (Early, Midday, and Late), and again across two time points (Morning and Afternoon). We also investigated whether activity pattern, sex (as a control variable), and other traits associated with gut passage rate (diet, body mass) could help predict the presence and timing of circadian patterns of fecal glucocorticoid metabolites across mammal species. We found that fecal glucocorticoid metabolite levels in E. rubriventer fluctuate throughout the day, with lowest levels in the morning and peak levels in the afternoon. None of the variables that we tested predicted whether daily fecal glucocorticoid metabolites changed significantly throughout the day, nor when levels were likely to peak, across species. We stress the importance of controlling for sampling time and reporting these results as standard practice in studies of fecal glucocorticoid metabolites.

Mechanisms of the different effects of sucrose, glucose, fructose, and a glucose-fructose mixture on wheat starch gelatinization, pasting, and retrogradation.

The gelatinization, pasting, and retrogradation of starch influence texture, quality, and shelf-life attributes of many foods. The purpose of this work was to document the effects of a 50:50 glucose:fructose (glc:fru) mixture and sucrose solutions on these starch traits to provide a fundamental basis to explain the different texture and shelf-life attributes of baked goods formulated with these sugars. Differential scanning calorimetry, rapid visco analyzer, and oscillatory rheometry were used to quantify the effects of glucose, fructose, glc:fru mixture, and sucrose at different concentrations (0% to 60% w/w), on the gelatinization temperature, pasting, and retrogradation properties of wheat starch. Distinct differences were found between the effects of sucrose and those of the monosaccharides including the glc:fru mixture. Sucrose elevated Tgel and pasting temperature most and decreased other RVA parameters compared to the monosaccharides as concentration increased. Fructose and the glc:fru mixture promoted amylopectin retrogradation, while retrogradation was inhibited in sucrose and glucose solutions. The glc:fru mixture had similar effects on starch properties compared to fructose under static measurement conditions (DSC), and the effects were in between those of glucose and fructose under dynamic conditions when shear was applied (RVA and rheology). These effects are explained by the phase separation and/or solute partitioning of the monosaccharide constituents of the glc:fru mixture. Sugar solution physicochemical properties correlated strongly with starch gelatinization and retrogradation. The results substantiate the important relationship between sugar physicochemical properties and solution dynamics with starch thermal properties, which in turn affect the texture and structure of starch-containing food products. PRACTICAL APPLICATION: The quality attributes of starch-containing baked goods are influenced by how different amounts and types of sugars affect starch cooking properties. The underlying mechanisms of the different sugar effects involve solution viscosity, intermolecular hydrogen bonding, and phase separation. Substituting one sugar for another has less effect on these starch properties in products with lower sugar concentrations than in products with more sugar. Mixtures of sugars behave differently than single sugars in different conditions due to phase separation. Baked goods made with glucose:fructose mixtures in place of sucrose likely have higher amounts of gelatinized starch and increased firmness (i.e., staling or retrogradation) over time.

From hybrid to fully remote clinical trial amidst the COVID-19 pandemic: Strategies to promote recruitment, retention, and engagement in a randomized mHealth trial.

Clinical trials worldwide were disrupted when the COVID-19 pandemic began in early 2020. Most intervention trials moved to some form of remote implementation due to restrictions on in-person research activities. Although the proportion of remote trials is growing, they remain the vast minority of studies in part due to few successful examples. Our team transitioned Goals for Reaching Optimal Wellness (GROWell), an NIH-funded (R01NR017659) randomized control trial (RCT; ClinicalTrials.gov identifier NCT04449432) originally designed as a hybrid intervention, into a fully remote clinical trial. GROWell is a digital dietary intervention for people who enter pregnancy with overweight or obesity. Primary outcomes include gestational weight gain and six-month postpartum weight retention. Strategies that we have tested, refined, and deployed include: (a) use of a HIPAA-compliant, web-based participant recruitment and engagement platform; (b) use of a HIPAA-compliant digital health platform to disseminate GROWell and conduct study visits (c) interconnectivity of these two platforms for seamless recruitment, consent, enrollment, intervention delivery, follow-up, and study team blinding; (d) detailed SMS messages to address initial challenges with protocol adherence; (e) email notifications alerting the study team about missed participant surveys so they can follow-up; (f) remuneration using email gift cards with recipient choice of vendor; and (g) geotargeting social media campaigns to improve participation of Black Indigenous and People of Color Communities. These strategies have resulted in screen failure rates improving by 7%, study task adherence improving by an average of 20-30% across study visits, and study completion rates of 82%. Researchers may consider some or all of these approaches in future remote mHealth trials.

Women's status, breastfeeding support, and breastfeeding practices in the United States.

The objective of this study is to examine associations between state-level breastfeeding support and breastfeeding practices, controlling for women's status, in the U.S. We used publicly available data on state-level breastfeeding practices and support (international board-certified lactation consultants (IBCLC), births in Baby-Friendly hospitals, and La Leche League Leaders) for births in 2015 from the CDC Breastfeeding Report Card (2018) and other CDC reported data, and indicators of women's status from the Institute for Women's Policy Research reports (2015). We conducted an ecological study to estimate incidence rate ratios of exclusive breastfeeding at six months and breastfeeding at 12 months with breastfeeding supports using bivariate and multivariable Poisson regression. Political participation, poverty, and employment and earnings were associated with breastfeeding practices, as was each breastfeeding support in bivariate analyses. After controlling for women's status, only IBCLCs were positively associated with rates of exclusive breastfeeding at 6 months and continued breastfeeding at 12 months. For every additional IBCLC per 1000 live births, the rate of exclusive breastfeeding at 6 months increased by 5 percent (95% CI 1.03, 1.07) and the rate of breastfeeding at 12 months increased by 4 percent (95% CI 1.02, 1.06). Political participation, poverty, and employment and earnings were associated with breastfeeding practices, indicating a relationship between women's political and economic status and their breastfeeding practices in the U.S. Given the influence of women's status, increasing the number of IBCLCs may improve breastfeeding practices.

Automated prediction of site and sequence of protein modification with ATRP initiators.

One of the most straightforward and commonly used chemical modifications of proteins is to react surface amino groups (lysine residues) with activated esters. This chemistry has been used to generate protein-polymer conjugates, many of which are now approved therapeutics. Similar conjugates have also been generated by reacting activated ester atom transfer polymerization initiators with lysine residues to create biomacromolecular initiators for polymerization reactions. The reaction between activated esters and lysine amino groups is rapid and has been consistently described in almost every publication on the topic as a "random reaction". A random reaction implies that every accessible lysine amino group on a protein molecule is equally reactive, and as a result, that the reaction is indiscriminate. Nonetheless, the literature contradicts itself by also suggesting that some lysine amino groups are more reactive than others (as a function of pKa, surface accessibility, temperature, and local environment). If the latter assumption is correct, then the outcome of these reactions cannot be random at all, and we should be able to predict the outcome from the structure of the protein. Predicting the non-random outcome of a reaction between surface lysines and reactive esters could transform the speed at which active bioconjugates can be developed and engineered. Herein, we describe a robust integrated tool that predicts the activated ester reactivity of every lysine in a protein, thereby allowing us to calculate the non-random sequence of reaction as a function of reaction conditions. Specifically, we have predicted the intrinsic reactivity of each lysine in multiple proteins with a bromine-functionalised N-hydroxysuccinimide initiator molecule. We have also shown that the model applied to PEGylation. The rules-based analysis has been coupled together in a single Python program that can bypass tedious trial and error experiments usually needed in protein-polymer conjugate design and synthesis.

Microtentacle Formation in Ovarian Carcinoma.

BACKGROUND: The development of chemoresistance to paclitaxel and carboplatin represents a major therapeutic challenge in ovarian cancer, a disease frequently characterized by malignant ascites and extrapelvic metastasis. Microtentacles (McTNs) are tubulin-based projections observed in detached breast cancer cells. In this study, we investigated whether ovarian cancers exhibit McTNs and characterized McTN biology. METHODS: We used an established lipid-tethering mechanism to suspend and image individual cancer cells. We queried a panel of immortalized serous (OSC) and clear cell (OCCC) cell lines as well as freshly procured ascites and human ovarian surface epithelium (HOSE). We assessed by Western blot ß-tubulin isotype, α-tubulin post-translational modifications and actin regulatory proteins in attached/detached states. We studied clustering in suspended conditions. Effects of treatment with microtubule depolymerizing and stabilizing drugs were described. RESULTS: Among cell lines, up to 30% of cells expressed McTNs. Four McTN morphologies (absent, symmetric-short, symmetric-long, tufted) were observed in immortalized cultures as well as ascites. McTN number/length varied with histology according to metastatic potential. Most OCCC overexpressed class III ß-tubulin. OCCC/OSC cell lines exhibited a trend towards more microtubule-stabilizing post-translational modifications of α-tubulin relative to HOSE. Microtubule depolymerizing drugs decreased the number/length of McTNs, confirming that McTNs are composed of tubulin. Cells that failed to form McTNs demonstrated differential expression of α-tubulin- and actin-regulating proteins relative to cells that form McTNs. Cluster formation is more susceptible to microtubule targeting agents in cells that form McTNs, suggesting a role for McTNs in aggregation. CONCLUSIONS: McTNs likely participate in key aspects of ovarian cancer metastasis. McTNs represent a new therapeutic target for this disease that could refine therapies, including intraperitoneal drug delivery.

North Carolina LGBTQ Domestic Violence Response Initiative: Building Capacity to Provide Safe, Affirming Services.

Intimate partner violence (IPV) is as prevalent or more so as reported by lesbian, gay, bisexual, transgender, and queer (LGBTQ) individuals compared with non-LGBTQ individuals. Yet largely due to prior harmful and nonaffirming experiences with service providers, they are reluctant to seek services from domestic violence service providers (DVSP). These factors, combined with the reality that those who do seek services may not be provided with safe, affirming, and effective responses, exacerbate the impact of IPV on LGBTQ health and recovery. The North Carolina (NC) LGBTQ Domestic Violence Response Initiative (NC Initiative) was developed to increase DVSP capacity to serve LGBTQ survivors. This article describes the first four phases of the Initiative's development: (1) formative focus group research with DVSPs statewide, identifying agencies' interest in improving their responses to LGBTQ survivors; (2) Initiative launch, including funding and identification of partners; (3) development of capacity assessment tools; and (4) review of baseline capacity assessment findings. The development of tools, specifically designed to assess capacity of DVSPs to serve LGBTQ survivors, is a unique contribution as no tools existed to do so. The common capacity-building needs and data-informed recommendations identified provide a starting point for agencies expanding into LGBTQ-focused work, who may not have the benefit of grant funding, tools, or evaluators to assist. Our findings, lessons learned, and tools may be used by agencies to evaluate and inform their own practices, thereby creating safer, more affirming agencies that are capable provide effective care to meet unique needs of LGBTQ survivors.

Goals for Reaching Optimal Wellness (GROWell): A clinical trial protocol of a digital dietary intervention for pregnant and postpartum people with prenatal overweight or obesity.

BACKGROUND: Excess gestational weight gain (EGWG) is associated with multiple pregnancy complications and health risks for birthing people and their infants. Likewise, postpartum weight retention (PPWR), or not losing all pregnancy weight, has long-term health consequences. EGWG among people who enter pregnancy with overweight or obesity have worse obstetric outcomes and increased PPWR compared to women who gain within Institute of Medicine guidelines. METHODS: This study protocol describes the details of a blinded, randomized clinical trial of GROWell: Goals for Reaching Optimal Wellness, a mHealth tool designed to improve diet quality among people who enter pregnancy with overweight or obese BMIs to help them achieve appropriate GWG and safe postpartum pregnancy weight loss. Individuals with overweight and obesity will be randomly assigned to an attention control or intervention arm. The intervention group will receive personalized, goal-oriented text messages regarding dietary choices, while the attention control group will receive text messages about healthy pregnancy, labor, delivery, and early infancy. Both groups will complete online surveys at baseline, follow up, 3 and 6 months postpartum. RESULTS AND DISCUSSION: Currently, 162 subjects have been enrolled. Outcomes associated with GWG and pregnancy are expected in late 2023, while outcomes on postpartum weight retention GROWell adherence are expected in late 2024. The results of this trial will support the use of an evidence-based mHealth tool to be integrated into clinical practice to reduce EGWG and PPWR among pregnant people with overweight and obese BMIs, a resource that is currently lacking. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT04449432. Registered on June 26, 2020.

The first responder exposure to contaminating powder on dog fur during intranasal and intramuscular naloxone administration.

OBJECTIVE: To determine whether first responders delivering naloxone by either the IM or intranasal (IN) route were at risk of contamination with inert powder simulating canine opioid exposure. DESIGN: Prospective, crossover design. SETTING: Research study (university setting). ANIMALS: Ten clinically normal working dogs ranging from 9 to 44 months were enrolled based on training to detect odor and ability to be restrained with minimal stress. All enrolled dogs completed both arms of the study without adverse effects. INTERVENTIONS: Dogs were randomly assigned to fentanyl reversal with either IM or IN naloxone and then the alternate treatment after a 7-day washout period. Prior to reversal, dogs' heads were brushed with an inert glow-in-the-dark powder. First responders (the same 2 individuals for all dogs) performing the reversal were photographed under ultraviolet light prior to and 5 min after administering the medication. Digital photographs were scored by body region for presence of glowing powder by observers blinded to timing of photograph (pre- or postreversal) and route of reversal (IM vs IN). MEASUREMENTS AND MAIN RESULTS: Compared to pretreatment, the inert powder scores were higher after treatment regardless of route of naloxone administration (P < 0.001). IN administration led to higher contamination than IM naloxone, particularly in the chest area (P = 0.012). CONCLUSIONS: Both IN and IM naloxone administration to dogs with clinical signs of opioid exposure result in a risk of first responders becoming contaminated with powder, which could include opioids. Awareness, proper personal protective equipment, and appropriate posttreatment decontamination are important to reduce risk of inadvertent exposure of mucous membranes to these contaminating powders.

REPRINT OF: Physical and Mental Health Effects of Intimate Partner Violence for Men and Women.

EDITOR'S NOTE: This article is a reprint of a previously published article. For citation purposes, please use the original publication details: Coker AL, Davis KE, Arias I, et al. Physical and mental health effects of intimate partner violence for men and women. Am J Prev Med. 1985;1(6):1-8. BACKGROUND: Few population-based studies have assessed the physical and mental health consequences of both psychological and physical intimate partner violence (IPV) among women or men victims. This study estimated IPV prevalence by type (physical, sexual, and psychological) and associated physical and mental health consequences among women and men. METHODS: The study analyzed data from the National Violence Against Women Survey (NVAWS) of women and men aged 18 to 65. This random-digit-dial telephone survey included questions about violent victimization and health status indicators. RESULTS: A total of 28.9% of 6790 women and 22.9% of 7122 men had experienced physical, sexual, or psychological IPV during their lifetime. Women were significantly more likely than men to experience physical or sexual IPV (relative risk [RR]=2.2, 95% confidence interval [CI]=2.1, 2.4) and abuse of power and control (RR=1.1, 95% CI=1.0, 1.2), but less likely than men to report verbal abuse alone (RR=0.8, 95% CI=0.7, 0.9). For both men and women, physical IPV victimization was associated with increased risk of current poor health; depressive symptoms; substance use; and developing a chronic disease, chronic mental illness, and injury. In general, abuse of power and control was more strongly associated with these health outcomes than was verbal abuse. When physical and psychological IPV scores were both included in logistic regression models, higher psychological IPV scores were more strongly associated with these health outcomes than were physical IPV scores. CONCLUSIONS: Both physical and psychological IPV are associated with significant physical and mental health consequences for both male and female victims.

Prospective evaluation of health status, quality of life and clinical outcomes following implantable defibrillator generator exchange.

BACKGROUND: Little is known about health status and quality of life (QoL) after implantable cardioverter-defibrillator (ICD) generator exchange (GE). METHODS: We prospectively followed patients undergoing first-time ICD GE. Serial assessments of health status were performed by administering the 36-Item Short Form Survey (SF-36). RESULTS: Mean age was 67.5 ± 14.3 years, left ventricle ejection fraction (LVEF) was 36.5% ± 15.0% and over 40% of the cohort had improved LVEF to > 35% at the time of GE. SF-36 scores were significantly worse in physical/general health domains compared to domains of emotional/social well-being ( P < 0.001 for each comparison). Physical health scores were significantly worse among those with medical comorbidities including diabetes, chronic obstructive pulmonary disease and atrial fibrillation. Mean follow-up was 1.6 ± 0.5 years after GE. Overall SF-36 scores remained stable across all domains during follow-up. Survival at 3 years post-GE was estimated at 80%. Five patients died during follow-up and most deaths were adjudicated as non-arrhythmic in origin. Four patients experienced appropriate ICD shocks after GE, three of whom had LVEF which remains impaired LVEF (i.e., < 35%) at the time of GE. CONCLUSION: Patients undergoing ICD GE have significantly worse physical health compared to emotional/social well-being, which is associated with the presence of medical comorbidities. In terms of clinical outcomes, the incidence of appropriate shocks after GE among those with improvement in LVEF is very low, and most deaths post-procedure appear to be non-arrhythmic in origin. These data represent an attempt to more fully characterize the spectrum of QoL and clinical outcomes after GE.

Efficacy of an mHealth Intervention (BRAVE) to Promote Mental Wellness for American Indian and Alaska Native Teenagers and Young Adults: Randomized Controlled Trial.

BACKGROUND: Culturally relevant interventions are needed to help American Indian and Alaska Native (AI/AN) teenagers and young adults navigate common risky situations involving family and friends, including drug and alcohol misuse, dating violence, and suicidality. Leveraging We R Native, a multimedia health resource for Native teenagers and young adults, staff of the Northwest Portland Area Indian Health Board designed the BRAVE intervention for Native youth. The program is delivered via SMS text messaging and includes role model videos, mental wellness strategies, links to culturally relevant resources, and social support from family and friends. OBJECTIVE: We aim to conduct a randomized controlled trial of the BRAVE intervention among AI/AN teenagers and young adults (aged 15-24 years) to assess its impact on their physical, mental, and spiritual health; their resilience and self-esteem; and their coping and help-seeking skills. METHODS: From October to December 2019, we recruited 2334 AI/AN teenagers and young adults nationwide via social media channels and SMS text messages and enrolled 1044 participants. AI/AN teenagers and young adults enrolled in the study received either BRAVE SMS text messages, designed to improve mental health, help-seeking skills, and cultural resilience, or 8 weeks of science, technology, engineering, and math (STEM) SMS text messages, designed to elevate and reaffirm Native voices in STEM and medicine and then received the BRAVE SMS text messages. The impacts of the BRAVE intervention were tested using linear mixed-effect models and linear regressions. RESULTS: A total of 833 AI/AN teenagers and young adults were included in the analysis. Individuals in the BRAVE and STEM arms showed significant positive trends over the course of the study for all outcomes, except cultural identity and help-seeking behavior. Mean scores were significantly different for health (P<.001), resilience (P<.001), negative coping (P=.03), positive coping (P<.001), self-efficacy (P=.02), and self-esteem (P<.001). Changes in help-seeking self-efficacy were significant in those exhibiting risky behaviors at baseline to exit (P=.01). Those who reported positive coping scores at baseline also reported better health on average; however, no difference was found in risky drug and alcohol use (P<.001). The number of participants who used SMS text messages to help themselves increased from 69.1% (427/618) at 3 months to 76% (381/501; P<.001) at 8 months. Similarly, the number of participants who used SMS text messages to help friends or family members increased from 22.4% (138/616) at 3 months to 54.6% (272/498) at 8 months. CONCLUSIONS: This is the first national randomized controlled trial of a mobile health intervention among AI/AN teenagers and young adults to test the efficacy of a mental wellness intervention in relation to STEM career messages. This study provides new insights for supporting the next generation of AI/AN changemakers. TRIAL REGISTRATION: ClinicalTrials.gov NCT04979481; https://clinicaltrials.gov/ct2/show/NCT04979481.

Blood Mercury in Three Populations of Endangered Whooping Crane (Grus americana).

We investigated concentrations of blood total mercury (THg) in three extant populations of endangered Whooping Crane (Grus americana). Blood THg was greater in cranes reintroduced during 2001-2008 that range in the eastern US (median = 0.31 ug/g ww) than both wild cranes in central North America (median = 0.11 ug/g ww) and from captivity (median = 0.01 ug/g ww). The median THg blood concentrations in the two free-ranging populations of Whooping Cranes were low compared to reproductive toxic thresholds suggested for other large bird species (> 4.3 ug/g ww), but a singular elevated concentration was observed in one crane (1.04 ug/g ww; 6% of samples from the reintroduced population). These results likely reflect variable dietary Hg exposure among these populations, but should prompt additional biomonitoring to discern risk of greater Hg exposure nearer to the time of breeding and potential effects on productivity.

The BRAVE Study: Formative Research to Design a Multimedia Intervention for American Indian and Alaska Native Young Adults.

American Indian and Alaska Native (AI/AN) young adults are strong and resilient. Interventions designed to improve their mental health and help-seeking skills are especially needed, particularly those that include culturally relevant resources and relatable role models. This paper presents formative research from the BRAVE study, a five-year community based participatory research project led by the Northwest Portland Area Indian Health Board. Formative research included three phases and more than 38 AI/AN young adults and content experts from across the United States. Results indicate that behavioral interventions can be feasibly delivered via text message to AI/AN young adults and that including Native youth in the formative research is critical to designing a comprehensive, culturally-responsive intervention. Lessons learned from this five-year process may help other youth-serving organizations, prevention programs, policymakers, researchers, and educators as they support the next generation of AI/AN leaders.

Dysregulated Provision of Oxidisable Substrates to the Mitochondria in ME/CFS Lymphoblasts.

Although understanding of the biomedical basis of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is growing, the underlying pathological mechanisms remain uncertain. We recently reported a reduction in the proportion of basal oxygen consumption due to ATP synthesis by Complex V in ME/CFS patient-derived lymphoblast cell lines, suggesting mitochondrial respiratory inefficiency. This was accompanied by elevated respiratory capacity, elevated mammalian target of rapamycin complex 1 (mTORC1) signaling activity and elevated expression of enzymes involved in the TCA cycle, fatty acid ß-oxidation and mitochondrial transport. These and other observations led us to hypothesise the dysregulation of pathways providing the mitochondria with oxidisable substrates. In our current study, we aimed to revisit this hypothesis by applying a combination of whole-cell transcriptomics, proteomics and energy stress signaling activity measures using subsets of up to 34 ME/CFS and 31 healthy control lymphoblast cell lines from our growing library. While levels of glycolytic enzymes were unchanged in accordance with our previous observations of unaltered glycolytic rates, the whole-cell proteomes of ME/CFS lymphoblasts contained elevated levels of enzymes involved in the TCA cycle (p = 1.03 × 10-4), the pentose phosphate pathway (p = 0.034, G6PD p = 5.5 × 10-4), mitochondrial fatty acid ß-oxidation (p = 9.2 × 10-3), and degradation of amino acids including glutamine/glutamate (GLS p = 0.034, GLUD1 p = 0.048, GOT2 p = 0.026), branched-chain amino acids (BCKDHA p = 0.028, BCKDHB p = 0.031) and essential amino acids (FAH p = 0.036, GCDH p = 0.006). The activity of the major cellular energy stress sensor, AMPK, was elevated but the increase did not reach statistical significance. The results suggest that ME/CFS metabolism is dysregulated such that alternatives to glycolysis are more heavily utilised than in controls to provide the mitochondria with oxidisable substrates.

Changes in membrane elasticity caused by the hydrophobic surfactant proteins correlate poorly with adsorption of lipid vesicles.

To establish how the hydrophobic surfactant proteins, SP-B and SP-C, promote adsorption of lipids to an air/water interface, we used X-ray diffuse scattering (XDS) to determine an order parameter of the lipid chains (Sxray) and the bending modulus of the lipid bilayers (KC). Samples contained different amounts of the proteins with two sets of lipids. Dioleoylphosphatidylcholine (DOPC) provided a simple, well characterized model system. The nonpolar and phospholipids (N&PL) from extracted calf surfactant provided the biological mix of lipids. For both systems, the proteins produced changes in Sxray that correlated well with KC. The dose-response to the proteins, however, differed. Small amounts of protein generated large decreases in Sxray and KC for DOPC that progressed monotonically. The changes for the surfactant lipids were erratic. Our studies then tested whether the proteins produced correlated effects on adsorption. Experiments measured the initial fall in surface tension during adsorption to a constant surface area, and then expansion of the interface during adsorption at a constant surface tension of 40 mN m-1. The proteins produced a sigmoidal increase in the rate of adsorption at 40 mN m-1 for both lipids. The results correlated poorly with the changes in Sxray and KC in both cases. Disordering of the lipid chains produced by the proteins, and the softening of the bilayers, fail to explain how the proteins promote adsorption of lipid vesicles.

Characterization of ultrasound-mediated delivery of trastuzumab to normal and pathologic spinal cord tissue.

Extensive studies on focused ultrasound (FUS)-mediated drug delivery through the blood-brain barrier have been published, yet little work has been published on FUS-mediated drug delivery through the blood-spinal cord barrier (BSCB). This work aims to quantify the delivery of the monoclonal antibody trastuzumab to rat spinal cord tissue and characterize its distribution within a model of leptomeningeal metastases. 10 healthy Sprague-Dawley rats were treated with FUS + trastuzumab and sacrificed at 2-h or 24-h post-FUS. A human IgG ELISA (Abcam) was used to measure trastuzumab concentration and a 12 ± fivefold increase was seen in treated tissue over control tissue at 2 h versus no increase at 24 h. Three athymic nude rats were inoculated with MDA-MB-231-H2N HER2 + breast cancer cells between the meninges in the thoracic region of the spinal cord and treated with FUS + trastuzumab. Immunohistochemistry was performed to visualize trastuzumab delivery, and semi-quantitative analysis revealed similar or more intense staining in tumor tissue compared to healthy tissue suggesting a comparable or greater concentration of trastuzumab was achieved. FUS can increase the permeability of the BSCB, improving drug delivery to specifically targeted regions of healthy and pathologic tissue in the spinal cord. The achieved concentrations within the healthy tissue are comparable to those reported in the brain.