Chinese hamster ovary cell line engineering strategies for modular production of custom extracellular vesicles.

Continuously secreted by all cell types, extracellular vesicles (EVs) are small membrane-bound structures which shuttle bioactive cargo between cells across their external environment. Their central role as natural molecular messengers and ability to cross biological barriers has garnered significant attention in the use of EVs as therapeutic delivery vehicles. Still, harnessing the potential of EVs is faced with many obstacles. A cell line engineering approach can be used to exploit EVs to encapsulate a bespoke cargo of interest. However, full details regarding native EV-loading mechanisms remain under debate, making this a challenge. While Chinese hamster ovary (CHO) cells are well known to be the preferred host for recombinant therapeutic protein production, their application as an EV producer cell host has been largely overlooked. In this study, we engineered CHO DG44 cells to produce custom EVs with bespoke cargo. To this end, genetic constructs employing split green fluorescent protein technology were designed for tagging both CD81 and protein cargoes to enable EV loading via self-assembling activity. To demonstrate this, NanoLuc and mCherry were used as model reporter cargoes to validate engineered loading into EVs. Experimental findings indicated that our custom EV approach produced vesicles with up to 15-fold greater cargo compared with commonly used passive loading strategies. When applied to recipient cells, we observed a dose-dependent increase in cargo activity, suggesting successful delivery of engineered cargo via our custom CHO EVs.

Changes in mixed ethnicity households and neighbourhood transitions in England and Wales.

Conventional explanations of neighbourhood ethnic transitions consider what drives differential growth in ethnic group populations without regard to household composition. We enrich these nonhousehold approaches by using consistent Census data on neighbourhoods and households for England and Wales for 2001, 2011 and 2021 to analyse connections between mixed-ethnicity households and neighbourhood ethnic diversity. We employ a neighbourhood typology of ethnic diversity that identifies neighbourhoods as either low- or moderate-diversity, or high-diversity, where no single ethnic group is in the majority. We focus particularly on White-majority and highly diverse neighbourhoods given the dominance of the former in residential spaces in England and Wales, and because they are the principal source of transitions to highly diverse neighbourhoods. Mixed-ethnicity households have become an increasingly important feature of the ethnic diversification of England and Wales; by 2021, almost 15% of multiperson households were mixed, a growth from 12% in 2011 and 9% in 2001. We find that higher levels of neighbourhood ethnic diversity are associated with higher shares of mixed-ethnicity households. In high-diversity neighbourhoods, for example, around 30% of multiperson households (nearly a quarter of a million households) were ethnically mixed in 2021. Levels of household mixing in moderate-diversity White neighbourhoods were considerably higher than in low-diversity White neighbourhoods. The same is found for diversifying neighbourhoods. Neighbourhoods that become more ethnically diverse were typically home to higher rates of mixed-ethnicity households. Stably moderately diverse White neighbourhoods also had a higher proportion of mixed households. Studies of neighbourhood transition that focus on individuals in neighbourhoods may be missing important processes operating in the intimate spaces of the home. Including this intermediate scale of analysis adds to our understanding of neighbourhood ethnic mixing and processes of integration.

Pneumococcal Disease Outbreak at a State Prison, Alabama, USA, September 1-October 10, 20181.

A pneumococcal disease outbreak caused by Streptococcus pneumoniae serotype 12F occurred in a state prison in Alabama, USA. Among 1,276 inmates, 40 cases were identified (3 confirmed, 2 probable, 35 suspected). Close living quarters, substance use, and underlying conditions likely contributed to disease risk. Prophylaxis for close contacts included azithromycin and 23-valent pneumococcal polysaccharide vaccine.

A comparative analysis of recombinant Fab and full-length antibody production in Chinese hamster ovary cells.

Monoclonal antibodies are the leading class of biopharmaceuticals in terms of numbers approved for therapeutic purposes. Antigen-binding fragments (Fab) are also used as biotherapeutics and used widely in research applications. The dominant expression systems for full-length antibodies are mammalian cell-based, whereas for Fab molecules the preference has been an expression in bacterial systems. However, advances in CHO and downstream technologies make mammalian systems an equally viable option for small- and large-scale Fab production. Using a panel of full-length IgG antibodies and their corresponding Fab pair with different antigen specificities, we investigated the impact of the IgG and Fab molecule format on production from Chinese hamster ovary (CHO) cells and assessed the cellular capability to process and produce these formats. The full-length antibody format resulted in the recovery of fewer mini-pools posttransfection when compared to the corresponding Fab fragment format that could be interpreted as indicative of a greater overall burden on cells. Antibody-producing cell pools that did recover were subsequently able to achieve higher volumetric protein yields (mg/L) and specific productivity than the corresponding Fab pools. Importantly, when the actual molecules produced per cell of a given format was considered (as opposed to mass), CHO cells produced a greater number of Fab molecules per cell than obtained with the corresponding IgG, suggesting that cells were more efficient at making the smaller Fab molecule. Analysis of cell pools showed that gene copy number was not correlated to the subsequent protein production. The amount of mRNA correlated with secreted Fab production but not IgG, whereby posttranscriptional processes act to limit antibody production. In summary, we provide the first comparative description of how full-length IgG and Fab antibody formats impact on the outcomes of a cell line construction process and identify potential limitations in their production that could be targeted for engineering increases in the efficiency in the manufacture of these recombinant antibody formats.

The evolution and stability of multi-ethnic residential neighbourhoods in England.

This paper analyses the most ethnically diverse spaces in England. We define multi-ethnic neighbourhoods as spaces where no one group is in a majority and at least five ethnic groups have representation. Around 4% of all English neighbourhoods (Lower Layer Super Output Areas) met these criteria in 2011. Often mislabelled as "segregated" spaces, the growth of ethnically diverse neighbourhoods helps benchmark increased inter-ethnic contact, yet we know very little about their spatial extent and the dynamics of their expansion. We use Census data for 1991, 2001, and 2011 to consider how neighbourhood-level diversity has changed during a period of substantial increase in ethnic diversity at the national scale. To what extent did these highly diverse areas grow, and what is the geography of that growth? Which types of areas did these neighbourhoods transition from? For example, were multi-ethnic neighbourhoods formerly low or moderately diverse, and which groups dominated these locales? We also consider if multi-ethnic neighbourhoods are here to stay, or if they are compositionally unstable. We reveal a surprising aspect in England's neighbourhood transitions: multi-ethnic neighbourhoods are highly stable, and increasingly so. Some 88% of neighbourhoods that were multi-ethnic in 1991 retained their high-diversity status in 2001, while over 95% of 2001 multi-ethnic neighbourhoods remained highly diverse by 2011. This is a different story to that of the USA, where high-diversity neighbourhoods have received more scholarly attention, and where these neighbourhoods have high attrition rates, functioning as stepping stones to another type of space. We explore the demographic and housing dynamics associated with this stability.

Mixed measures: different definitions of racially diverse neighborhoods compared.

Focusing on neighborhoods that researchers consider particularly diverse, this paper assesses the ways scholars have characterized neighborhood racial diversity in the United States. Social scientists use a variety of methods to define and measure highly racially diverse places, resulting in a single label being used to capture very different aspects of a census tract's racial demography. We examine the criteria used to classify neighborhood racial diversity to bring perspective on the logic behind various approaches. We then group the range of schemas into several broad types from which we choose a representative four. These form the basis for a series of empirical comparisons using U.S. Census data to reveal the contexts where the taxonomies produce similar outcomes and those where they do not. The analysis goes on to consider the implications stemming from the choices social scientists make when they opt for one approach over another.

Wilderness Medical Society Clinical Practice Guidelines for the Prevention and Treatment of Heat Illness: 2019 Update.

The Wilderness Medical Society convened an expert panel in 2011 to develop a set of evidence-based guidelines for the recognition, prevention, and treatment of heat illness. We present a review of the classifications, pathophysiology, and evidence-based guidelines for planning and preventive measures, as well as best practice recommendations for both field- and hospital-based therapeutic management of heat illness. These recommendations are graded based on the quality of supporting evidence and balance the benefits and risks or burdens for each modality. This is an updated version of the original Wilderness Medical Society Practice Guidelines for the Treatment and Prevention of Heat-Related Illness published in 2013.

Where science, technology, engineering, and mathematics (STEM) graduates move: Human capital, employment patterns, and interstate migration in the United States.

This research investigates the interstate migration of workers in the United States who have earned an undergraduate STEM (science, technology, engineering, and mathematics) degree compared with those who have not. We build on previous studies that (a) classified "skilled" workers as having earned an undergraduate degree (b) used net migration gain or loss as a yardstick of relative destination attraction, and (c) advanced the idea that physical amenities play an outsized role in labour market preferences for skilled workers. We calibrate the attractivity of states for three levels of human capital and then evaluate these assessments of relative attractivity to show that workers with different types of human capital respond to different labour market signals in contradictory ways. Amenity, measured by heating degree days, has little to do with the state-to-state migration of workers who have a STEM degree, yet helps explain the migration patterns of workers with no undergraduate degree. Employment growth in a state influences migration for degreed workers in the recessionary years but not in the period of recovery. The opposite holds for workers without a degree. States with high percentages of any type of degreed workers attract both STEM and non-STEM degreed migrants but not those without a degree. States with a large share of STEM degreed workers in their degreed workforce are especially attractive for STEM degreed migrants. The conclusions discuss what the findings imply about diverging access to labour market opportunity by human capital and state higher education policy.

Racial and Ethnic Disparities in Travel for Head and Neck Cancer Treatment and the Impact of Travel Distance on Survival.

BACKGROUND: Patients who travel a long distance (≥50 miles) for cancer care have improved outcomes. However, to the authors' knowledge, the prevalence of long travel distances for treatment by patients with head and neck squamous cell carcinoma (HNSCC), and the effect of travel distance on overall survival (OS), remains unknown. METHODS: The authors used the National Cancer Data base from 2004 through 2013 to identify patients with HNSCC undergoing definitive treatment. Travel distance for treatment was categorized as short (<12.5 miles), intermediate (12.5-49.9 miles), and long (50-249.9 miles). The primary outcome, OS, was evaluated using Cox shared-frailty modeling. A secondary outcome, factors associated with intermediate and long travel distances, was evaluated using multivariable hierarchical logistic regression. RESULTS: Among 118,000 patients with HNSCC, 62,753 (53.2%), 40,644 (34.4%), and 14,603 (12.4%) patients, respectively, traveled short, intermediate, and long distances for treatment. After adjusting for relevant covariates, long travel distance was associated with treatment at academic and high-volume centers. Patients of black race, of Hispanic ethnicity, with Medicaid insurance, and who were treated with nonsurgical treatment were less likely to travel long distances for treatment (P<.001). Traveling a long distance for treatment was associated with improved OS on multivariable analysis (adjusted hazard ratio, 0.93; 95% confidence interval, 0.89-0.96) compared with a short distance. CONCLUSIONS: Traveling a long distance for HNSCC treatment is associated with improved survival, especially for patients receiving nonsurgical management. Racial and ethnic disparities in travel for HNSCC treatment exist. As regionalization of care continues, future work should identify and address reasons for racial and ethnic disparities in travel that may prevent access to care at high-volume facilities. Cancer 2018;000:000-000. © 2018 American Cancer Society.

Transmitting and decoding facial expressions of emotion during healthy aging: More similarities than differences.

Older adults tend to perform more poorly than younger adults on emotional expression identification tasks. The goal of the present study was to test a processing mechanism that might explain these differences in emotion recognition-specifically, age-related variation in the utilization of specific visual cues. Seventeen younger and 17 older adults completed a reverse-correlation emotion categorization task (Bubbles paradigm), consisting of a large number of trials in each of which only part of the visual information used to convey an emotional facial expression was revealed to participants. The task allowed us to pinpoint the visual features each group used systematically to correctly recognize the emotional expressions shown. To address the possibility that faces of different age groups are differently processed by younger and older adults, we included younger, middle-aged, and older adult face models displaying happy, fearful, angry, disgusted, and sad facial expressions. Our results reveal strong similarity in the utilization of visual information by younger and older adult participants in decoding the emotional expressions from faces across ages-particularly for happy and fear emotions. These findings suggest that age-related differences in strategic information use are unlikely to contribute to the decline of facial expression recognition skills observed in later life.

Remaking White Residential Segregation: Metropolitan Diversity and Neighborhood Change in the United States.

Between 1990 and 2010, the white population share in US metropolitan areas fell from 73.5 percent to 60.3 percent. This paper explores how this decline affected the number and composition of white census tracts (tracts in which non-Latino whites constitute the largest group). In 1990, white tracts comprised 82 percent of all metropolitan tracts. By 2010, this percentage had fallen to 70 percent, paralleling the percentage point drop in white population share. This loss was concentrated among the most segregated white tracts - those with low diversity. White tracts that were moderately diverse actually doubled in number between 1990 and 2010 although this increase was insufficient to cancel the loss of low diversity white tracts. We model the effects of metropolitan characteristics on white-tract change by metropolitan area. Greater metropolitan-scale diversity increases the probability that low-diversity white tracts transition to moderate-diversity white. Moderately diverse white tracts, however, become more stable with increased diversity. A large metropolitan percentage of blacks or the foreign born, however, reverses this stabilizing effect, increasing the probability that moderately diverse white tracts transition to non-white tracts (i.e. where a non-white group is the largest group). Thus the level and composition of metropolitan diversity matters for the trajectory of moderately diverse white tracts. Overall, the formation of new white tracts, possibly the result of gentrification, coupled with the emergence of moderately diverse white tracts and an increasing share of whites living in such residential environments, suggests a reconfiguration rather than a dissolving of white dominated neighborhood space in response to increased diversity in surrounding metropolitan contexts.

A PROSPECTIVE ON ZELINSKY'S HYPOTHESIS OF THE MOBILITY TRANSITION.

Wilbur Zelinsky's 1971 paper in Geographical Review entitled the "Hypothesis of the Mobility Transition" was both forward-looking and offered innovative ideas regarding human geographic mobility. One of the most interesting aspects of the paper was a set of predictions for mobility in a "future superadvanced society". Many of these predictions have now come to pass, including a general decline in international and internal migration and residential change, the increasing regulation of migration - especially internally, and the possibility that the widespread adoption of information and communication technologies has impacted human geographic mobility. Hence, this essay looks at the mobility transition not as an obsolete frame of reference but as a prescient, pliable, and adaptable framework which not only informs the study of human geographic mobility today but also, perhaps, even into the future.

Effect of time to initiation of postoperative radiation therapy on survival in surgically managed head and neck cancer.

BACKGROUND: The objective of this study was to determine the effects of National Comprehensive Cancer Network (NCCN) guideline-adherent initiation of postoperative radiation therapy (PORT) and different time-to-PORT intervals on the overall survival (OS) of patients with head and neck squamous cell carcinoma (HNSCC). METHODS: The National Cancer Data Base was reviewed for the period of 2006-2014, and patients with HNSCC undergoing surgery and PORT were identified. Kaplan-Meier survival estimates, Cox regression analysis, and propensity score matching were used to determine the effects of initiating PORT within 6 weeks of surgery and different time-to-PORT intervals on survival. RESULTS: This study included 41,291 patients. After adjustments for covariates, starting PORT >6 weeks postoperatively was associated with decreased OS (adjusted hazard ratio [aHR], 1.13; 99% confidence interval [CI], 1.08-1.19). This finding remained in the propensity score-matched subset (hazard ratio, 1.21; 99% CI, 1.15-1.28). In comparison with starting PORT 5 to 6 weeks postoperatively, initiating PORT earlier was not associated with improved survival (aHR for ≤ 4 weeks, 0.93; 99% CI, 0.85-1.02; aHR for 4-5 weeks, 0.92; 99% CI, 0.84-1.01). Increasing durations of delay beyond 7 weeks were associated with small, progressive survival decrements (aHR, 1.09, 1.10, and 1.12 for 7-8, 8-10, and >10 weeks, respectively). CONCLUSIONS: Nonadherence to NCCN guidelines for initiating PORT within 6 weeks of surgery was associated with decreased survival. There was no survival benefit to initiating PORT earlier within the recommended 6-week timeframe. Increasing durations of delay beyond 7 weeks were associated with small, progressive survival decrements. Cancer 2017;123:4841-50. © 2017 American Cancer Society.

The Matching of STEM Degree Holders with STEM Occupations in Large Metropolitan Labor Markets in the United States.

Workers in science, technology, engineering, and mathematics (STEM) are vital for regional and national prosperity. Not every person with STEM qualifications, however, finds employment in a STEM job. This article analyzes the geography of this matching between STEM degree holders and certain types of STEM occupations across large metropolitan labor markets in the United States. We find that although labor-market size has no effect, living in denser STEM labor markets elevates the probabilities of matching; having an advanced degree enhances this effect. Women are also far less likely to be matched than men; being black or Latino additionally lowers the chances of matching. Combining spatial effects with individual attributes increases probabilities of matching in places with high concentrations of STEM jobs for women, racial minorities, and the foreign born, but these advantages are often the same for white, native-born men. In denser STEM labor markets the job-matching advantage spans the labor pool, conferring no differential benefit for different population subgroups.

Influence of a metatarsus adductus foot type on plantar pressures during walking in adults using a pedobarograph.

Metatarsus adductus is a relatively common congenital foot deformity that is often unrecognized at birth. Thus, the adult foot with metatarsus adductus is prone to pathologic entities that have been theorized to result from lateral column overload. We present a descriptive study comparing plantar foot pressure distribution during gait in subjects with and without metatarsus adductus. A total of 65 subjects were recruited for the study: 28 subjects with and 37 subjects without metatarsus adductus. An EMED(®) pedobarograph was used to collect the data. The analysis of the peak pressure and pressure-time integral in each of the 8 regions of the plantar surface of the foot showed significant (p < .05) differences between each of the regions and a significant (p < .05) interaction effect between the 8 regions and the 2 groups. A series of independent Student's t tests were therefore performed to determine which of the plantar regions showed a significant difference between the 2 groups. The result of those t tests showed that the peak pressure and pressure-time integral were significantly different (p < .05) between the 2 groups for the "heel," "lateral midfoot," and "lateral forefoot." The results of the present study support the concept that during gait, the adult foot with metatarsus adductus has increased peak plantar pressures on the lateral side of the foot.

A prospective randomized blister prevention trial assessing paper tape in endurance distances (Pre-TAPED).

OBJECTIVE: Friction foot blisters are a common injury occurring in up to 39% of marathoners, the most common injury in adventure racing, and represent more than 70% of medical visits in multi-stage ultramarathons. The goal of the study was to determine whether paper tape could prevent foot blisters in ultramarathon runners. METHODS: This prospective randomized trial was undertaken during RacingThePlanet 155-mile (250-km), 7-day self-supported ultramarathons in China, Australia, Egypt, Chile, and Nepal in 2010 and 2011. Paper tape was applied prerace to one randomly selected foot, with the untreated foot acting as the own control. The study end point was development of a hot spot or blister on any location of either foot. RESULTS: One hundred thirty-six participants were enrolled with 90 (66%) having completed data for analysis. There were 36% women, with a mean age of 40 ± 9.4 years (range, 25-40 years) and pack weight of 11 ± 1.8 kg (range, 8-16 kg). All participants developed blisters, with 89% occurring by day 2 and 59% located on the toes. No protective effect was observed by the intervention (47 versus 35; 52% versus 39%; P = .22), with fewer blisters occurring around the tape on the experimental foot than under the tape (23 vs 31; 25.6% versus 34.4%), yet 84% of study participants when queried would choose paper tape for blister prevention in the future. CONCLUSIONS: Although paper tape was not found to be significantly protective against blisters, the intervention was well tolerated with high user satisfaction.

Wilderness Medical Society practice guidelines for the prevention and treatment of heat-related illness: 2014 update.

The Wilderness Medical Society (WMS) convened an expert panel to develop a set of evidence-based guidelines for the recognition, prevention, and treatment of heat illness. We present a review of the classifications, pathophysiology, and evidence-based guidelines for planning and preventive measures as well as best practice recommendations for both field and hospital-based therapeutic management of heat illness. These recommendations are graded on the basis of the quality of supporting evidence, and balance between the benefits and risks or burdens for each modality. This is an updated version of the original WMS Practice Guidelines for the Prevention and Treatment of Heat-Related Illness published in Wilderness & Environmental Medicine 2013;24(4):351-361.

The allure of new immigrant destinations and the Great Recession in the United States.

In the 1990s, the immigrant population in the United States dispersed to non-traditional settlement locations (what have become known as "new immigrant destinations"). This paper examines whether the allure of new destinations persisted in the 2000s with a particular focus on the internal migration of the foreign born during the recent deep recessionary period and its aftermath. Three specific questions motivate the analysis. First, are immigrants, much like the US-born population, becoming less migratory within the country over time? Second, is immigrant dispersal from traditional gateways via internal migration continuing despite considerable economic contraction in many new destination metropolitan areas? Third, is immigration from aboard a substitute for what appears to be declining immigrant internal migration to new destinations? The findings reveal a close correlation between the declining internal migration propensity of the US-born and immigrants in the last two decades. We also observe parallels between the geographies of migration of native- and foreign-born populations with both groups moving to similar metropolitan areas in the 1990s. This redistributive association, however, weakened in the subsequent decade as new destination metropolitan areas lost their appeal for both groups, especially immigrants. There is no evidence to suggest that immigration from abroad is substituting for the decline in immigrant redistribution through internal migration to new destinations. Across destination types the relationship between immigration from abroad and the internal migration of the foreign born remained the same before, during, and after the Great Recession.

The migration response to the Legal Arizona Workers Act.

The 2008 Legal Arizona Workers Act (LAWA) requires all public and private employers to authenticate the legal status of their workers using the federal employment verification system known as E-Verify. With LAWA, Arizona became the first state to have a universal mandate for employment verification. While LAWA targets unauthorized workers, most of whom are Latino immigrants, other groups could experience LAWA's effects, such as those who share households with undocumented workers. In addition, employers may seek to minimize their risk of LAWA penalties by not hiring those who appear to them as more likely to be unauthorized, such as naturalized Latino immigrants and US-born Latinos. Existing research has found a reduction in foreign-born Latino employment and population in response to LAWA. This paper asks a different question: have groups that are most likely to be affected by the law migrated to other states? We find a significant and sustained increase in the internal outmigration rate from Arizona of foreign-born, noncitizen Latinos - the group most likely to include the unauthorized - after the passage of LAWA. There was no significant LAWA internal migration response by foreign-born Latino citizens. US-born Latinos showed some signs of a LAWA-induced internal migration response after the law went into effect, but it is not sustained. The results indicate that local and state immigration policy can alter the settlement geography of the foreign born. This leads us to speculate about how immigrant settlement may adjust in the coming years to the intersecting geographies of post-recession economic opportunity and tiered immigration policies.

Wilderness Medical Society practice guidelines for the prevention and treatment of heat-related illness.

The Wilderness Medical Society (WMS) convened an expert panel to develop a set of evidence-based guidelines for the recognition, prevention, and treatment of heat-related illness. We present a review of the classifications, pathophysiology, and evidence-based guidelines for planning and preventive measures as well as best-practice recommendations for both field- and hospital-based therapeutic management of heat-related illness. These recommendations are graded based on the quality of supporting evidence and the balance between the benefits and risks or burdens for each modality.