Adaptive DNA amplification of synthetic gene circuit opens a way to overcome cancer chemoresistance.

Drug resistance continues to impede the success of cancer treatments, creating a need for experimental model systems that are broad, yet simple, to allow the identification of mechanisms and novel countermeasures applicable to many cancer types. To address these needs, we investigated a set of engineered mammalian cell lines with synthetic gene circuits integrated into their genome that evolved resistance to Puromycin. We identified DNA amplification as the mechanism underlying drug resistance in 4 out of 6 replicate populations. Triplex-forming oligonucleotide (TFO) treatment combined with Puromycin could efficiently suppress the growth of cell populations with DNA amplification. Similar observations in human cancer cell lines suggest that TFOs could be broadly applicable to mitigate drug resistance, one of the major difficulties in treating cancer.

Overnight Melatonin Concentration and Sleep Quality Are Associated with the Clinical Features of Polycystic Ovary Syndrome.

Circulating melatonin is elevated in women with polycystic ovary syndrome (PCOS); whether circadian disruptions coincide with sleep disturbances in women with PCOS or their symptom severity is unclear. The objective of this observational pilot study was to determine whether altered patterns of melatonin excretion are associated with reduced sleep quality in women with versus without PCOS. Participants underwent a clinical assessment, transvaginal ultrasound, and reproductive hormone testing. Morning and evening urine samples were assayed for urinary 6-sulfatoxymelatonin (MEL) as a proxy for melatonin production. The night (morning MEL)-to-day (evening MEL) ratio, or N:D ratio, was determined to approximate the rhythm of MEL production. Sleep quality and duration were assessed using the Pittsburgh Sleep Quality Index (PSQI) and wrist actigraphy. No differences were detected in overnight MEL, daytime MEL, or the N:D ratio in participants with PCOS versus controls. The PCOS group experienced reduced weekend sleep efficiency vs. controls (81% vs. 88% p < 0.05). The number of follicles per ovary (FNPO) was positively associated with overnight MEL (r = 0.359, p < 0.05). Weekend sleep time and overnight MEL concentrations were dependent on PCOS status. Therefore, diagnostic features of PCOS were associated with MEL production and sleep disturbances, suggesting that women with a more severe clinical presentation of PCOS may be more likely to experience altered MEL production or sleep disturbances.

Notch1 cortical signaling regulates epithelial architecture and cell-cell adhesion.

Notch receptors control tissue morphogenic processes that involve coordinated changes in cell architecture and gene expression, but how a single receptor can produce these diverse biological outputs is unclear. Here, we employ a 3D model of a human ductal epithelium to reveal tissue morphogenic defects result from loss of Notch1, but not Notch1 transcriptional signaling. Instead, defects in duct morphogenesis are driven by dysregulated epithelial cell architecture and mitogenic signaling which result from the loss of a transcription-independent, Notch1 cortical signaling mechanism that ultimately functions to stabilize adherens junctions and cortical actin. We identify that Notch1 localization and cortical signaling are tied to apical-basal cell restructuring and discover that a Notch1-FAM83H interaction underlies control of epithelial adherens junctions and cortical actin. Together, these results offer new insights into Notch1 signaling and regulation and advance a paradigm in which transcriptional and cell adhesive programs might be coordinated by a single receptor.

Rationale and design of the SenseWhy project: A passive sensing and ecological momentary assessment study on characteristics of overeating episodes.

Objectives: Overeating interventions and research often focus on single determinants and use subjective or nonpersonalized measures. We aim to (1) identify automatically detectable features that predict overeating and (2) build clusters of eating episodes that identify theoretically meaningful and clinically known problematic overeating behaviors (e.g., stress eating), as well as new phenotypes based on social and psychological features. Method: Up to 60 adults with obesity in the Chicagoland area will be recruited for a 14-day free-living observational study. Participants will complete ecological momentary assessments and wear 3 sensors designed to capture features of overeating episodes (e.g., chews) that can be visually confirmed. Participants will also complete daily dietitian-administered 24-hour recalls of all food and beverages consumed. Analysis: Overeating is defined as caloric consumption exceeding 1 standard deviation of an individual's mean consumption per eating episode. To identify features that predict overeating, we will apply 2 complementary machine learning methods: correlation-based feature selection and wrapper-based feature selection. We will then generate clusters of overeating types and assess how they align with clinically meaningful overeating phenotypes. Conclusions: This study will be the first to assess characteristics of eating episodes in situ over a multiweek period with visual confirmation of eating behaviors. An additional strength of this study is the assessment of predictors of problematic eating during periods when individuals are not on a structured diet and/or engaged in a weight loss intervention. Our assessment of overeating episodes in real-world settings is likely to yield new insights regarding determinants of overeating that may translate into novel interventions.

Case report: Clinical and pathological findings in a canine patient with intervertebral disk extrusion resembling progressive myelomalacia.

A 4-year-old female spayed dog presented to the emergency department for non-ambulatory tetraparesis, which progressed to tetraplegia. Computed tomography (CT) confirmed cervical intervertebral disk extrusion at C5-6 extending to C6-7, and an emergency ventral slot was performed. After the procedure, the patient was placed on mechanical ventilation due to respiratory failure. Repeat assessment upon weaning her ventilatory support suggested the patient's neurological status had declined. Based on her deterioration and suspicion of progressive myelomalacia on magnetic resonance imaging (MRI), she was euthanized. Post-mortem histopathology of the spinal cord supported the presence of progressive myelomalacia. To the author's knowledge, this is the first case report describing a progressive myelomalacia in a canine patient with cervical intervertebral disk extrusion.

Adherens junctions organize size-selective proteolytic hotspots critical for Notch signalling.

Adherens junctions (AJs) create spatially, chemically and mechanically discrete microdomains at cellular interfaces. Here, using a mechanogenetic platform that generates artificial AJs with controlled protein localization, clustering and mechanical loading, we find that AJs also organize proteolytic hotspots for γ-secretase with a spatially regulated substrate selectivity that is critical in the processing of Notch and other transmembrane proteins. Membrane microdomains outside of AJs exclusively organize Notch ligand-receptor engagement (LRE microdomains) to initiate receptor activation. Conversely, membrane microdomains within AJs exclusively serve to coordinate regulated intramembrane proteolysis (RIP microdomains). They do so by concentrating γ-secretase and primed receptors while excluding full-length Notch. AJs induce these functionally distinct microdomains by means of lipid-dependent γ-secretase recruitment and size-dependent protein segregation. By excluding full-length Notch from RIP microdomains, AJs prevent inappropriate enzyme-substrate interactions and suppress spurious Notch activation. Ligand-induced ectodomain shedding eliminates size-dependent segregation, releasing Notch to translocate into AJs for processing by γ-secretase. This mechanism directs radial differentiation of ventricular zone-neural progenitor cells in vivo and more broadly regulates the proteolysis of other large cell-surface receptors such as amyloid precursor protein. These findings suggest an unprecedented role of AJs in creating size-selective spatial switches that choreograph γ-secretase processing of multiple transmembrane proteins regulating development, homeostasis and disease.

Small-molecule allosteric inhibitors of GPX4.

Encouraged by the dependence of drug-resistant, metastatic cancers on GPX4, we examined biophysical mechanisms of GPX4 inhibition, which revealed an unexpected allosteric site. We found that this site was involved in native regeneration of GPX4 under low glutathione conditions. Covalent binding of inhibitors to this allosteric site caused a conformational change, inhibition of activity, and subsequent cellular GPX4 protein degradation. To verify this site in an unbiased manner, we screened a library of compounds and identified and validated that an additional compound can covalently bind in this allosteric site, inhibiting and degrading GPX4. We determined co-crystal structures of six different inhibitors bound in this site. We have thus identified an allosteric mechanism for small molecules targeting aggressive cancers dependent on GPX4.

Coronary Artery Restenosis in Women by History of Preeclampsia.

Background A history of preeclampsia is associated with increased risk of coronary artery disease and experimental evidence suggests that a history of preeclampsia also increases the risk of restenosis. However, the extent to which a history of preeclampsia is associated with risk of restenosis after percutaneous coronary intervention in women is unknown. Methods and Results We included 6065 parous women aged ≤65 years with first percutaneous coronary intervention on 9452 segments 2006 to 2017, linking nationwide data on percutaneous coronary intervention and delivery history in Sweden. Main outcomes were clinical restenosis and target lesion revascularization within 2 years. We accounted for segment-, procedure-, and patient-related potential predictors of restenosis in proportional hazards regression models. Restenosis occurred in 345 segments (3.7%) and target lesion revascularization was performed on 383 patients (6.3%). A history of preeclampsia was neither significantly associated with risk of restenosis (predictor-accounted hazard ratio [HR], 0.71 [95% CI, 0.41-1.23]) nor target lesion revascularization (0.74 [95% CI, 0.51-1.07]) compared with a normotensive pregnancy history. When term and preterm preeclampsia were investigated separately, segments in women with a history of term preeclampsia had a lower risk of restenosis (predictor-accounted HR, 0.45 [95% CI, 0.21-0.94]). A history of preeclampsia was not significantly associated with death by any cause within 2 years of the index procedure (predictor-accounted HR 1.06, [95% CI, 0.62-1.80]). Conclusions A history of preeclampsia was not associated with increased risk of restenosis but instead some evidence pointed to a decreased risk. To facilitate future studies and allow for replication, concomitant collection of data on pregnancy complication history and percutaneous coronary intervention outcomes in women is warranted.

Hexose translocation mediated by SlSWEET5b is required for pollen maturation in Solanum lycopersicum.

Pollen fertility is critical for successful fertilization and, accordingly, for crop yield. While sugar unloading affects the growth and development of all types of sink organs, the molecular nature of sugar import to tomato (Solanum lycopersicum) pollen is poorly understood. However, sugar will eventually be exported transporters (SWEETs) have been proposed to be involved in pollen development. Here, reverse transcription-quantitative polymerase chain reaction (PCR) revealed that SlSWEET5b was markedly expressed in flowers when compared to the remaining tomato SlSWEETs, particularly in the stamens of maturing flower buds undergoing mitosis. Distinct accumulation of SlSWEET5b-ß-glucuronidase activities was present in mature flower buds, especially in anther vascular and inner cells, symplasmic isolated microspores (pollen grains), and styles. The demonstration that SlSWEET5b-GFP fusion proteins are located in the plasma membrane supports the idea that the SlSWEET5b carrier functions in apoplasmic sugar translocation during pollen maturation. This is consistent with data from yeast complementation experiments and radiotracer uptake, showing that SlSWEET5b operates as a low-affinity hexose-specific passive facilitator, with a Km of ∼36 mM. Most importantly, RNAi-mediated suppression of SlSWEET5b expression resulted in shrunken nucleus-less pollen cells, impaired germination, and low seed yield. Moreover, stamens from SlSWEET5b-silenced tomato mutants showed significantly lower amounts of sucrose (Suc) and increased invertase activity, indicating reduced carbon supply and perturbed Suc homeostasis in these tissues. Taken together, our findings reveal the essential role of SlSWEET5b in mediating apoplasmic hexose import into phloem unloading cells and into developing pollen cells to support pollen mitosis and maturation in tomato flowers.

Food Handling Concerns and Practices at Home during the COVID-19 Pandemic by Food Security Status.

ABSTRACT: There is limited examination about coronavirus disease 19 (COVID-19)-related food handling concerns and practices that cause chemical or microbial contamination and illness, particularly among those with food insecurity. We investigated consumer food handling concerns and practices during the COVID-19 pandemic and whether they differed by food insecurity status. An online survey was distributed among Chicago, IL, residents between 15 July and 21 August 2020 (n = 437). Independent t tests and Fisher's exact tests were used to identify differences in food handling concerns and practices between those with and without food insecurity (alpha = 0.05). Survey items included questions about food handling practices that were considered safe or neutral (i.e., washing hands and produce with water, sanitizing food packaging) and unsafe (i.e., using cleaning agents to wash foods, leaving perishable foods outside) by using 5-point Likert-style scales or categorical responses (i.e., yes, no). Participant responses fell between "slightly" and "somewhat" concerned about contracting COVID-19 from food and food packaging (mean ± standard error [SE]: 2.7 ± 0.1). Although participants reported washing their hands before eating and before preparing foods at least "most of the time" (mean ± SE: 4.4 ± 0.0 and 4.5 ± 0.0, respectively), only one-third engaged in unsafe practices. The majority of participants (68%) indicated that they altered food handling practices due to the COVID-19 pandemic and received information about food safety from social media (61%). When investigating differences in concerns and practices by food insecurity status, food insecure participants were more concerned about COVID-19 foodborne transmission for all food items (all P < 0.001) and more frequently performed unsafe food handling practices than those with food security (all P < 0.001). Results from this study suggest more investigation is needed to understand barriers to safe food handling knowledge and practices, particularly among those with food insecurity.

Comparative Validity of Mostly Unprocessed and Minimally Processed Food Items Differs Among Popular Commercial Nutrition Apps Compared with a Research Food Database.

BACKGROUND: Commercial nutrition apps are increasingly used to evaluate diet. Evaluating the comparative validity of nutrient data from commercial nutrition app databases is important to determine the merits of using these apps for dietary assessment. OBJECTIVE: Nutrient data from four commercial nutrition apps were compared with a research-based food database, Nutrition Data System for Research (NDSR) (version 2017). DESIGN: Comparative validation study. PARTICIPANTS/SETTING: An investigator identified the 50 most frequently consumed foods (22% of total reported foods) from a weight-loss study in Chicago, IL, during 2017. Nutrient data were compared between four commercial databases with NDSR. MAIN OUTCOME MEASURES: Comparative validity of energy, macronutrients, and other nutrient data (ie, total sugars, fiber, saturated fat, cholesterol, calcium, and sodium). STATISTICAL ANALYSES PERFORMED: Intraclass correlation coefficients (ICCs) evaluated agreement between commercial databases with the NDSR for foods that were primarily un- and minimally processed and by the three most frequently consumed food groups. Bland-Altman plots determined degree of bias for calories between commercial databases and NDSR. RESULTS: This study observed excellent agreement between NDSR and CalorieKing (ICC range = 0.90 to 1.00). Compared with NDSR, agreement for Lose It! and MyFitnessPal ranged from good to excellent (ICC range = 0.89 to 1.00), with the exception of fiber in MyFitnessPal (ICC = 0.67). Fitbit showed the widest variability with NDSR (ICC range = 0.52 to 0.98). When evaluating by food group, Fitbit had poor agreement for all food groups, with the lowest agreement observed for fiber within the vegetable group (ICC = 0.16). Bland-Altman plots confirmed ICC energy results but also found that MyFitnessPal had the poorest agreement to NDSR (mean 8.35 [SD 133.31] kcal) for all food items. CONCLUSIONS: Degree of agreement varied by commercial nutrition app. CalorieKing and Lose It! had mostly excellent agreement with NDSR for all investigated nutrients. Fitbit showed the widest variability in agreement with NDSR for most nutrients, which may reflect how well the app can accurately capture diet.

Association of number of bites and eating speed with energy intake: Wearable technology results under free-living conditions.

Personalized weight management strategies are gaining interest. However, knowledge is limited regarding eating habits and association with energy intake, and current technologies limit assessment in free-living situations. We assessed associations between eating behavior and time of day with energy intake using a wearable camera under free-living conditions and explored if obesity modifies the associations. Sixteen participants (50% with obesity) recorded free-living eating behaviors using a wearable fish-eye camera for 14 days. Videos were viewed by trained annotators who confirmed number of bites, eating speed, and time of day for each eating episode. Energy intake was determined by a trained dietitian performing 24-h diet recalls. Greater number of bites, reduced eating speed, and increased BMI significantly predicted higher energy intake among all participants (P < 0.05, each). There were no significant interactions between obesity and number of bites, eating speed, or time of day (p > 0.05). Greater number of bites and reduced eating speed were significantly associated with higher energy intake in participants without obesity. Results show that under free-living conditions, more bites and slower eating speed predicted higher energy intake when examining consumption of foods with beverages. Obesity did not modify these associations. Findings highlight how eating behaviors can impact energy balance and can inform weight management interventions using wearable technology.

Using optogenetics to link myosin patterns to contractile cell behaviors during convergent extension.

Distinct patterns of actomyosin contractility are often associated with particular epithelial tissue shape changes during development. For example, a planar-polarized pattern of myosin II localization regulated by Rho1 signaling during Drosophila body axis elongation is thought to drive cell behaviors that contribute to convergent extension. However, it is not well understood how specific aspects of a myosin pattern influence the multiple cell behaviors, including cell intercalation, cell shape changes, and apical cell area fluctuations, that simultaneously occur during morphogenesis. Here, we developed two optogenetic tools, optoGEF and optoGAP, to activate or deactivate Rho1 signaling, respectively. We used these tools to manipulate myosin patterns at the apical side of the germband epithelium during Drosophila axis elongation and analyzed the effects on contractile cell behaviors. We show that uniform activation or inactivation of Rho1 signaling across the apical surface of the germband is sufficient to disrupt the planar-polarized pattern of myosin at cell junctions on the timescale of 3-5 min, leading to distinct changes in junctional and medial myosin patterns in optoGEF and optoGAP embryos. These two perturbations to Rho1 activity both disrupt axis elongation and cell intercalation but have distinct effects on cell area fluctuations and cell packings that are linked with changes in the medial and junctional myosin pools. These studies demonstrate that acute optogenetic perturbations to Rho1 activity are sufficient to rapidly override the endogenous planar-polarized myosin pattern in the germband during axis elongation. Moreover, our results reveal that the levels of Rho1 activity and the balance between medial and junctional myosin play key roles not only in organizing the cell rearrangements that are known to directly contribute to axis elongation but also in regulating cell area fluctuations and cell packings, which have been proposed to be important factors influencing the mechanics of tissue deformation and flow.

Obesity Status on associations between cancer-related beliefs and health behaviors in cancer survivors: Implications for patient-clinician communication.

OBJECTIVE: Associations between cancer beliefs and health behavior engagement are largely unexplored in cancer survivors, particularly among those with overweight and obesity. We investigated belief-behavior associations for cancer survivors, and whether obesity altered these associations. METHODS: Cancer survivors were identified from the National Cancer Institute HINTS Survey 5 data and classified as having had an obesity-related cancer or not. Linear and multiple logistic regression analyses examined whether cancer risk beliefs and self-efficacy predicted dining out behaviors and physical activity (PA). Restricted analyses were conducted in those with overweight or obesity. RESULTS: Low self-efficacy to take care of one's health was associated with longer sitting time in the overall sample (p = 0.04). In cancer survivors with overweight or obesity, engagement in healthier behaviors was associated with 1) feeling less overwhelmed by cancer risk recommendations and 2) believing that PA or obesity influences cancer development (both p < 0.05). Among those with overweight and obesity, associations between cancer beliefs and health behaviors were not significantly different by cancer type (obesity-related vs. not). CONCLUSIONS: Obesity altered associations between cancer risk beliefs and health behavior engagement from the overall sample. PRACTICE IMPLICATIONS: Weight status may be a useful tailoring factor when delivering health-promoting interventions for cancer survivors.

Associations of diet, physical activity and polycystic ovary syndrome in the Coronary Artery Risk Development in Young Adults Women's Study.

BACKGROUND: Current evidence supports the adoption of healthy diet and physical activity (PA) behaviors in patients with polycystic ovary syndrome (PCOS), given the positive effects of those behaviors on physical well-being. An improved understanding of the associations between diet and PA with PCOS is needed to ascertain whether tailored dietary and PA recommendations are needed for this population. Thus, we investigated the associations of diet and PA with PCOS and its isolated features. METHODS: Cross-sectional study. Of the 748 women who were included in this study from the Coronary Artery Risk Development in Young Adults (CARDIA) Women's Study, 40 were classified as having PCOS, 104 had isolated hyperandrogenism (HA) and 75 had isolated oligomenorrhea (OA). Dietary intake was measured using the CARDIA diet history questionnaire and diet quality was scored using the Alternative Healthy Eating Index 2010; a higher score indicated a better quality diet. Self-reported PA was measured using a validated interviewer-administered questionnaire. Polytomous logistic regression analyses examined the associations between diet and PA with PCOS, HA, and OA status (outcomes), adjusting for age, race, total energy intake, education, and/or body mass index. The threshold for statistical significance was set at p < 0.05. RESULTS: Mean age of the participants was 25.4 years (SD 3.6) and 46.8% of participants were Black women. There was little to no association of total energy intake, nutrients, diet quality, and PA with PCOS, HA or OA status. CONCLUSION: Energy intake, nutrient composition, diet quality, and PA were not associated with PCOS, supporting recent PCOS guidelines of using national recommendations for the general population to encourage health-promoting behaviors among women with PCOS. However, longitudinal studies evaluating changes in diet and physical activity in relation to the development and/or the progression of PCOS are needed to establish a causal association.

eHealth Practices in Cancer Survivors With BMI in Overweight or Obese Categories: Latent Class Analysis Study.

BACKGROUND: eHealth technologies have been found to facilitate health-promoting practices among cancer survivors with BMI in overweight or obese categories; however, little is known about their engagement with eHealth to promote weight management and facilitate patient-clinician communication. OBJECTIVE: The objective of this study was to determine whether eHealth use was associated with sociodemographic characteristics, as well as medical history and experiences (ie, patient-related factors) among cancer survivors with BMI in overweight or obese categories. METHODS: Data were analyzed from a nationally representative cross-sectional survey (National Cancer Institute's Health Information National Trends Survey). Latent class analysis was used to derive distinct classes among cancer survivors based on sociodemographic characteristics, medical attributes, and medical experiences. Logistic regression was used to examine whether class membership was associated with different eHealth practices. RESULTS: Three distinct classes of cancer survivors with BMI in overweight or obese categories emerged: younger with no comorbidities, younger with comorbidities, and older with comorbidities. Compared to the other classes, the younger with comorbidities class had the highest probability of identifying as female (73%) and Hispanic (46%) and feeling that clinicians did not address their concerns (75%). The older with comorbidities class was 6.5 times more likely than the younger with comorbidities class to share eHealth data with a clinician (odds ratio [OR] 6.53, 95% CI 1.08-39.43). In contrast, the younger with no comorbidities class had a higher likelihood of using a computer to look for health information (OR 1.93, 95% CI 1.10-3.38), using an electronic device to track progress toward a health-related goal (OR 2.02, 95% CI 1.08-3.79), and using the internet to watch health-related YouTube videos (OR 2.70, 95% CI 1.52-4.81) than the older with comorbidities class. CONCLUSIONS: Class membership was associated with different patterns of eHealth engagement, indicating the importance of tailored digital strategies for delivering effective care. Future eHealth weight loss interventions should investigate strategies to engage younger cancer survivors with comorbidities and address racial and ethnic disparities in eHealth use.

Obesity, Insulin Resistance, and Hyperandrogenism Mediate the Link between Poor Diet Quality and Ovarian Dysmorphology in Reproductive-Aged Women.

The relationship between diet quality and ovarian morphology has biological plausibility yet remains unclear and was therefore evaluated. In a multicenter cross-sectional analysis, four dietary patterns were scored for 111 consecutive reproductive-aged women (18-45 years) using (1) Healthy Eating Index (HEI-2015); (2) alternative HEI-2010; (3) alternate Mediterranean Diet (aMED); (4) and Dietary Approaches to Stop Hypertension (DASH) indices. Ovarian volume (OV) and follicle number per ovary (FNPO) were evaluated on transvaginal ultrasonography. Relationships between dietary and ovarian morphology indices were evaluated by linear regression and mediation analyses. Associations between aMED and DASH scores and OV/FNPO were completely mediated by obesity, insulin resistance, and hyperandrogenism (All: p < 0.05), unlike direct associations (All: p ≥ 0.89). Namely, a 1-standard deviation [SD] increase in aMED score was associated with decreases in OV (0.09 SD; 0.4 mL) through reducing waist circumference. Likewise, a 1 SD increase in aMED and DASH score was associated with decreases in OV (0.07 SD; 0.3 mL) by reducing glucose response to a 75 g glucose tolerance test. A 1 SD increase in DASH score was associated with decreased FNPO (0.07 SD; 2 follicles) by reducing free androgen index (All: p < 0.05). Adherence to aMED and DASH eating plans was indirectly associated with significant improvements in ovarian form, providing novel mechanistic insights for future interventions about contributions of diet quality on ovarian function.

Effects of Autoclaving, EtOH, and UV Sterilization on the Chemical, Mechanical, Printability, and Biocompatibility Characteristics of Alginate.

Sterilization is a necessary step during the processing of biomaterials, but it can affect the materials' functional characteristics. This study characterizes the effects of three commonly used sterilization processes-autoclaving (heat-based), ethanol (EtOH; chemical-based), and ultraviolet (UV; radiation-based)-on the chemical, mechanical, printability, and biocompatibility properties of alginate, a widely used biopolymer for drug delivery, tissue engineering, and other biomedical applications. Sterility assessment tests showed that autoclaving was effective against Gram-positive and Gram-negative bacteria at loads up to 108 CFU/mL, while EtOH was the least effective. Nuclear magnetic-resonance spectroscopy showed that the sterilization processes did not affect the monomeric content in the alginate solutions. The differences in compressive stiffness of the three sterilized hydrogels were also not significant. However, autoclaving significantly reduced the molecular weight and polydispersity index, as determined via gel permeation chromatography, as well as the dynamic viscosity of alginate. Printability analyses showed that the sterilization process as well as the extrusion pressure and speed affected the number of discontinuities and spreading ratio in printed and cross-linked strands. Finally, human adipose-derived stem cells demonstrated over 90% viability in all sterilized hydrogels over 7 days, but the differences in cellular metabolic activity in the three groups were significant. Taken together, the autoclaving process, while demonstrating broad spectrum sterility effectiveness, also resulted in most notable changes in alginate's key properties. In addition to the specific results with the three sterilization processes and alginate, this study serves as a roadmap to characterize the interrelationships between sterilization processes, fundamental chemical properties, and resulting functional characteristics and processability of hydrogels.

Derivation of a clinical decision instrument to identify patients with status epilepticus who require emergent brain CT.

BACKGROUND: Studies have shown the value of CT brain imaging in adults with first-time seizures, but there are no recommendations regarding emergent brain CTs in persons with an established seizure disorders. Our study aimed to derive a clinical decision instrument (CDI) to determine which patients with status epilepticus (SE) require emergent brain imaging. METHODS: This was a retrospective chart review of patients who presented to our emergency department with SE between 2010 and 2018. Patients with first-time seizures were excluded. A priori, we defined high risk criteria for emergent imaging as well as positive findings on brain CT. High risk criteria included known malignancy, trauma, and immunosuppression. Positive CT scans included findings such as intracranial hemorrhage (ICH) and mass. RESULTS: We identified 214 patients who met inclusion criteria Of the 181 patients without high risk criteria, 3% had positive CT scans. Of the 33 patients with high risk criteria, 10% had positive CT scans. The sensitivity, specificity, PPV, and NPV for our initial CDI were 38%, 85%, 9%, and 97%. Adding the criterion of prior ICH would have lowered our miss rate to 0.6%. Modifying our CDI to 1) History of ICH, 2) Malignancy, 3) Immunosuppression, and 4) Trauma would result in a CDI with sensitivity, specificity, PPV, and NPV of 87.5%, 87.4%, 21.2%, and 99.5%. CONCLUSIONS: By using four criteria to identify high risk patients, we can defer CT scanning in the vast majority of patients with SE and known seizure disorders. This CDI should be prospectively validated before adoption.

Counting Bites With Bits: Expert Workshop Addressing Calorie and Macronutrient Intake Monitoring.

BACKGROUND: Conventional diet assessment approaches such as the 24-hour self-reported recall are burdensome, suffer from recall bias, and are inaccurate in estimating energy intake. Wearable sensor technology, coupled with advanced algorithms, is increasingly showing promise in its ability to capture behaviors that provide useful information for estimating calorie and macronutrient intake. OBJECTIVE: This paper aimed to summarize current technological approaches to monitoring energy intake on the basis of expert opinion from a workshop panel and to make recommendations to advance technology and algorithms to improve estimation of energy expenditure. METHODS: A 1-day invitational workshop sponsored by the National Science Foundation was held at Northwestern University. A total of 30 participants, including population health researchers, engineers, and intervention developers, from 6 universities and the National Institutes of Health participated in a panel discussing the state of evidence with regard to monitoring calorie intake and eating behaviors. RESULTS: Calorie monitoring using technological approaches can be characterized into 3 domains: (1) image-based sensing (eg, wearable and smartphone-based cameras combined with machine learning algorithms); (2) eating action unit (EAU) sensors (eg, to measure feeding gesture and chewing rate); and (3) biochemical measures (eg, serum and plasma metabolite concentrations). We discussed how each domain functions, provided examples of promising solutions, and highlighted potential challenges and opportunities in each domain. Image-based sensor research requires improved ground truth (context and known information about the foods), accurate food image segmentation and recognition algorithms, and reliable methods of estimating portion size. EAU-based domain research is limited by the understanding of when their systems (device and inference algorithm) succeed and fail, need for privacy-protecting methods of capturing ground truth, and uncertainty in food categorization. Although an exciting novel technology, the challenges of biochemical sensing range from a lack of adaptability to environmental effects (eg, temperature change) and mechanical impact, instability of wearable sensor performance over time, and single-use design. CONCLUSIONS: Conventional approaches to calorie monitoring rely predominantly on self-reports. These approaches can gain contextual information from image-based and EAU-based domains that can map automatically captured food images to a food database and detect proxies that correlate with food volume and caloric intake. Although the continued development of advanced machine learning techniques will advance the accuracy of such wearables, biochemical sensing provides an electrochemical analysis of sweat using soft bioelectronics on human skin, enabling noninvasive measures of chemical compounds that provide insight into the digestive and endocrine systems. Future computing-based researchers should focus on reducing the burden of wearable sensors, aligning data across multiple devices, automating methods of data annotation, increasing rigor in studying system acceptability, increasing battery lifetime, and rigorously testing validity of the measure. Such research requires moving promising technological solutions from the controlled laboratory setting to the field.