Using a Consensus-Driven Approach to Incorporate Holistic Review Into an Internal Medicine Residency Program.

Background: Despite growing interest in incorporating holistic review within residency admissions, implementation by residency programs remains challenging. Objective: To incorporate holistic review into the internal medicine residency program at the University of Wisconsin and to report initial feasibility and acceptability data. Methods: During the 2020-2021 application cycle, residency stakeholders performed a consensus-driven process to identify highly valued applicant attributes. We used a holistic review process to identify the presence of these attributes among applicants and updated our rank list algorithm to incorporate these attributes. We modified our interview screening criteria and rank list algorithm to de-emphasize other metrics. We surveyed stakeholders to assess time required for this process and compared our final rank list to what it would have been using our prior system. Results: The final list of 10 prioritized applicant attributes included extraordinary leadership, community service, and grit, among others. Among 25 matched residents, 8 (32%) were recognized to have exceptional achievement within one of these 10 attributes. Four members of the incoming intern class (16%) would have been in a rank position lower than our historical matched resident cutoff had they not received additional points for these attributes. Faculty reported that holistic review of applications took an additional 3.8 minutes on average. It was felt that current application materials limit the ability to implement a fully holistic review. Conclusions: The addition of holistic review to our residency admissions process was achieved using a consensus-driven approach and showed favorable feasibility and acceptability data.

A Sequence- and Ligation-Independent Cloning (SLIC) Procedure for the Insertion of Genes into a Plasmid Vector.

Molecular cloning is a routine technique for many laboratories with applications from genetic engineering to recombinant protein expression. While restriction-ligation cloning can be slow and inefficient, ligation-independent cloning uses long single-stranded overhangs generated by T4 DNA polymerase's 3' exonuclease activity to anneal the insert and plasmid vector prior to transformation. This chapter describes a fast, high-efficiency protocol for inserting one or more genes into a vector using sequence- and ligation-independent cloning (SLIC).

Predicted wind and solar energy expansion has minimal overlap with multiple conservation priorities across global regions.

Protected areas and renewable energy generation are critical tools to combat biodiversity loss and climate change, respectively. Over the coming decades, expansion of the protected area network to meet conservation objectives will be occurring alongside rapid deployment of renewable energy infrastructure to meet climate targets, driving potential conflict for a finite land resource. Renewable energy infrastructure can have negative effects on wildlife, and co-occurrence may mean emissions targets are met at the expense of conservation objectives. Here, we assess current and projected overlaps of wind and solar photovoltaic installations and important conservation areas across nine global regions using spatially explicit wind and solar data and methods for predicting future renewable expansion. We show similar levels of co-occurrence as previous studies but demonstrate that once area is accounted for, previous concerns about overlaps in the Northern Hemisphere may be largely unfounded, although they are high in some biodiverse countries (e.g., Brazil). Future projections of overlap between the two land uses presented here are generally dependent on priority threshold and region and suggest the risk of future conflict can be low. We use the best available data on protected area degradation to corroborate this level of risk. Together, our findings indicate that while conflicts between renewables and protected areas inevitably do occur, renewables represent an important option for decarbonization of the energy sector that would not significantly affect area-based conservation targets if deployed with appropriate policy and regulatory controls.

Randomized, double-blind, placebo-controlled trial of ISC 17536, an oral inhibitor of transient receptor potential ankyrin 1, in patients with painful diabetic peripheral neuropathy: impact of preserved small nerve fiber function.

ABSTRACT: Patients with chronic pain syndromes, such as those with painful peripheral neuropathy due to diabetes mellitus, have limited treatment options and suffer ongoing attrition of their quality of life. Safer and more effective treatment options are needed. One therapeutic approach encompasses phenotypic characterization of the neuropathic pain subtype, combined with the selection of agents that act on relevant mechanisms. ISC 17536 is a novel, orally available inhibitor of the widely expressed pain receptor, transient receptor potential ankyrin 1, which mediates nociceptive signaling in peripheral small nerve fibers. In this randomized, placebo-controlled, proof-of-concept trial, we assessed the safety and efficacy of 28-day administration of ISC 17536 in 138 patients with chronic, painful diabetic peripheral neuropathy and used quantitative sensory testing to characterize the baseline phenotype of patients. The primary end point was the change from baseline to end of treatment in the mean 24-hour average pain intensity score based on an 11-point pain intensity numeric rating scale. The study did not meet the primary end point in the overall patient population. However, statistically significant and clinically meaningful improvement in pain were seen with ISC 17536 in an exploratory hypothesis-generating subpopulation of patients with preserved small nerve fiber function defined by quantitative sensory testing. These results may provide a mechanistic basis for targeted therapy in specific pain phenotypes in line with current approaches of "precision medicine" or personalized pain therapeutics. The hypothesis is planned to be tested in a larger phase 2 study.

Effects of Soybean Cultivar, Foliar Application of Azoxystrobin, and Year on Seed Vigor and Microflora Under Delayed Harvest Conditions.

The effects of cultivar and foliar fungicide applications on soybean seed germination, vigor, microflora, and yield after delayed harvest were determined at the University of Arkansas Vegetable Research Station in Kibler, AR, from 2008 to 2010. Seven cultivars with varying levels of resistance to Diaporthe spp. or Cercospora spp. were treated or not treated with a foliar application of azoxystrobin at the R5 growth stage. Plots were harvested 3 weeks after the plants had reached harvest maturity. Yields were recorded, samples of seed were collected, and standard germination (SG) and accelerated aging (AA) were assessed. Seeds were also assayed for infection by fungi on modified potato dextrose agar and by bacteria on nutrient agar. Seed vigor was significantly reduced by infection with Diaporthe spp., Fusarium spp., and Bacillus subtilis, but not with Cercospora spp. Cultivar had a significant impact on yield, seed vigor, and seed infection levels. The cultivar Osage had consistently high seed vigor and low overall seed infection incidence throughout the study. MO/PSD-0259, AG 4403, and UA 4805 also had relatively high seed vigor and low seed infection rates. PI 80837 had a low incidence of seed infection by Diaporthe spp. and Fusarium spp. in 2008 and 2010, but high levels in 2009, when environmental conditions were especially favorable for these pathogens. AP 350 and Suweon97 had relatively high seed infection incidences, particularly of Diaporthe spp. and Fusarium spp., and relatively low seed vigor. Application of the foliar fungicide azoxystrobin at the R5 growth stage significantly increased AA across years and cultivars and increased seed infection by Diaporthe spp. in 2009 across cultivars. There were significant negative correlations between yield and seed infection by Diaporthe spp. and Bacillus subtilis during 1 year and with Fusarium spp. during all 3 years. Overall, resistance to seed infection can persist even when harvest is delayed. In addition to Diaporthe spp., other seedborne pathogens may reduce seed vigor and yield.

The Effect of Overexpression of Lrp5 on the Temporomandibular Joint.

OBJECTIVE: The temporomandibular joint (TMJ) is a unique fibrocartilaginous joint that adapts to mechanical loading through cell signaling pathways such as the Wnt pathway. Increased expression of low-density lipoprotein receptor-related protein 5 (Lrp5), a co-receptor of the Wnt pathway, is associated with a high bone mass (HBM) phenotype. The objective of this study was to analyze the effect of overexpression of Lrp5 on the subchondral bone and cartilage of the TMJ in mice exhibiting the HBM phenotype. DESIGN: Sixteen-week-old Lrp5 knock-in transgenic mice carrying either the A214V (EXP-A) or G171V (EXP-G) missense mutations, and wildtype controls (CTRL) were included in this study. Fluorescent bone labels, calcein, alizarin complexone, and demeclocycline were injected at 3.5, 7.5, and 11.5 weeks of age, respectively. The left mandibular condyle was used to compare the subchondral bone micro-computed tomography parameters and the right TMJ was used for histological analyses. Cartilage thickness, matrix proteoglycan accumulation, and immunohistochemical localization of Lrp5 and sclerostin were compared between the groups. RESULTS: Subchondral bone volume (BV) and percent bone volume (BV/TV) were significantly increased in both EXP-A and EXP-G compared with CTRL (P < 0.05) whereas trabecular spacing (Tb.Sp) was decreased. Cartilage thickness, extracellular matrix production, and expression of Lrp5 and Sost were all increased in the experimental groups. The separation between the fluorescent bone labels indicated increased endochondral maturation between 3.5 and 7.5 weeks. CONCLUSIONS: These data demonstrate that Lrp5 overexpression leads to adaptation changes in the mandibular condylar cartilage of the TMJ to prevent cartilage degradation.

The effect of overexpression of Lrp5 on orthodontic tooth movement.

OBJECTIVE: To analyse the effect of gain-of-function mutations in the low-density lipoprotein receptor-related protein 5 (Lrp5) on orthodontic tooth movement (OTM). SETTING AND SAMPLE POPULATION: A split-mouth study design was utilized. Thirty-two male Lrp5-high bone mass (HBM) knock-in mice including A214V and G171V mutants (n = 16/group) and sixteen C57BL/6 wild-type (WT) mice were included in the study. MATERIALS AND METHODS: A mouse model of OTM was used for mesial movement of the maxillary first molar using a closed-coil nickel titanium (NiTi) spring attached between the molar and the incisors. After 21 days, the dissected maxillae were scanned for micro-computed tomography (micro-CT) analyses and embedded in methyl methacrylate and paraffin for histological staining and imaging. Histological analyses included immunohistochemistry for sclerostin (Sost), tartrate-resistant acid phosphatase (TRAP) staining for osteoclasts and fluorescent imaging. RESULTS: OTM in the A214V and G171V groups was significantly less than the WT group. Bone volume (BV), per cent bone volume (BV/TV) and trabecular thickness (Tb.Th) were significantly increased in both A241V and G171V animals compared to the WT animals. On the compression side, decreased osteoclast activity was seen in both A214V and G171V groups compared to the WT group. Fluorescent labelling demonstrated that the pattern of bone deposition in the A214V animals was periosteal whereas the G171V animals added bone endocortically. CONCLUSION: Gain-of-function mutations of Lrp5 decrease orthodontic tooth movement by increasing alveolar bone mass and reducing osteoclast-mediated bone resorption.

Identifying Agricultural Frontiers for Modeling Global Cropland Expansion.

The increasing expansion of cropland is major driver of global carbon emissions and biodiversity loss. However, predicting plausible future global distributions of croplands remains challenging. Here, we show that, in general, existing global data aligned with classical economic theories of expansion explain the current (1992) global extent of cropland reasonably well, but not recent expansion (1992-2015). Deviations from models of cropland extent in 1992 ("frontierness") can be used to improve global models of recent expansion, most likely as these deviations are a proxy for cropland expansion under frontier conditions where classical economic theories of expansion are less applicable. Frontierness is insensitive to the land cover dataset used and is particularly effective in improving models that include mosaic land cover classes and the largely smallholder-driven frontier expansion occurring in such areas. Our findings have important implications as the frontierness approach offers a straightforward way to improve global land use change models.

Impacts of rising temperatures and farm management practices on global yields of 18 crops.

Understanding the impact of changes in temperature and precipitation on crop yields is a vital step in developing policy and management options to feed the world. As most existing studies are limited to a few staple crops, we implemented global statistical models to examine the influence of weather and management practices on the yields of 18 crops, accounting for 70% of crop production by area and 65% by calorific intake. Focusing on the impact of temperature, we found considerable heterogeneity in the responses of yields across crops and countries. Irrigation was found to alleviate negative implications from temperature increases. Countries where increasing temperature causes the most negative impacts are typically the most food insecure, with the lowest calorific food supply and average crop yield. International action must be coordinated to raise yields in these countries through improvement and modernization of agricultural practices to counteract future adverse impacts of climate change.

The influence of the global electric power system on terrestrial biodiversity.

Given its total contribution to greenhouse gas emissions, the global electric power sector will be required to undergo a fundamental transformation over the next decades to limit anthropogenic climate change to below 2 °C. Implications for biodiversity of projected structural changes in the global electric power sector are rarely considered beyond those explicitly linked to climate change. This study uses a spatially explicit consumption-based accounting framework to examine the impact of demand for electric power on terrestrial vertebrate biodiversity globally. We demonstrate that the biodiversity footprint of the electric power sector is primarily within the territory where final demand for electric power resides, although there are substantial regional differences, with Europe displacing its biodiversity threat along international supply chains. The relationship between size of individual components of the electric power sector and threat to biodiversity indicates that a shift to nonfossil sources, such as solar and wind, could reduce pressures on biodiversity both within the territory where demand for power resides and along international supply chains. However, given the current levels of deployment of nonfossil sources of power, there is considerable uncertainty as to how the impacts of structural changes in the global electric power system will scale. Given the strong territorial link between demand and associated biodiversity impacts, development of strong national governance around the electric power sector represents a clear route to mitigate threats to biodiversity associated with efforts to decarbonize society over the coming century.

Collagenolytic matrix metalloproteinases antagonize proteinase-activated receptor-2 activation, providing insights into extracellular matrix turnover.

The collagenase subfamily of matrix metalloproteinases (MMPs) have important roles in the remodeling of collagenous matrices. The proteinase-activated receptor (PAR) family has a unique mechanism of activation requiring proteolysis of an extracellular domain forming a neo-N terminus that acts as a tethered ligand, a process that has been associated with the development of arthritis. Canonical PAR2 activation typically occurs via a serine proteinase at Arg36-Ser37, but other proteinases can cleave PARs downstream of the tethered ligand and "disarm" the receptor. To identify additional cleavage sites within PAR2, we synthesized a 42-amino-acid peptide corresponding to the extracellular region. We observed that all three soluble MMP collagenases, MMP-1, MMP-8, and MMP-13, cleave PAR2 and discovered a novel cleavage site (Ser37-Leu38). Metalloproteinases from resorbing bovine nasal cartilage and recombinant human collagenases could cleave a quenched fluorescent peptide mimicking the canonical PAR2 activation region, and kinetic constants were determined. In PAR2-overexpressing SW1353 chondrocytes, we demonstrated that the activator peptide SLIGKV-NH2 induces rapid calcium flux, inflammatory gene expression (including MMP1 and MMP13), and the phosphorylation of extracellular signal-regulated kinase (ERK) and p38 kinase. The corresponding MMP cleavage-derived peptide (LIGKVD-NH2) exhibited no canonical activation; however, we observed phosphorylation of ERK, providing evidence of biased agonism. Importantly, we demonstrated that preincubation with active MMP-1 reduced downstream PAR2 activation by a canonical activator, matriptase, but not SLIGKV-NH2 These results support a role for collagenases as proteinases capable of disarming PAR2, revealing a mechanism that suppresses PAR2-mediated inflammatory responses.

Osteoblast differentiation during orthodontic tooth movement.

OBJECTIVES: To analyse the expression of osteoblast differentiation markers and osteoclast activity in the periodontal ligament (PDL) following 2, 4 and 7 days of orthodontic tooth movement (OTM) in an animal model. SETTING AND SAMPLE POPULATION: Eighteen C57BL/6 wild-type mice. MATERIAL AND METHODS: For the OTM model, orthodontic force was applied to the maxillary right first molar using a closed-coil NiTi spring activated between the molar and incisors. The left side served as the control. Following OTM, the dissected tissues were scanned for micro-computed tomography (micro-CT) analysis and processed for histology. Histological stains included tartrate-resistant acid phosphatase (TRAP) staining for osteoclasts and immunohistochemistry for osteoblast markers alpha-smooth muscle actin (α-SMA), osteopontin (OP) and osteocalcin (OC). RESULTS: Micro-CT analysis showed increasing OTM on days 2, 4 and 7 days as well as decrease in bone volume and per cent bone volume at 4 and 7 days. Statistically significant increases in osteoblast marker expression were seen in all groups when compared to the control. TRAP expression was highest at 4 and 7 days, α-SMA was highest at 2 days and OP/OC was highest at 4 days. CONCLUSION: During OTM, proliferation of pre-osteoblasts peaks at 2 days while mineralization of the osteoid peaks at 4 days. The osteoclast response is delayed.

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms.

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

Grassland futures in Great Britain - Productivity assessment and scenarios for land use change opportunities.

To optimise trade-offs provided by future changes in grassland use intensity, spatially and temporally explicit estimates of respective grassland productivities are required at the systems level. Here, we benchmark the potential national availability of grassland biomass, identify optimal strategies for its management, and investigate the relative importance of intensification over reversion (prioritising productivity versus environmental ecosystem services). Process-conservative meta-models for different grasslands were used to calculate the baseline dry matter yields (DMY; 1961-1990) at 1km2 resolution for the whole UK. The effects of climate change, rising atmospheric [CO2] and technological progress on baseline DMYs were used to estimate future grassland productivities (up to 2050) for low and medium CO2 emission scenarios of UKCP09. UK benchmark productivities of 12.5, 8.7 and 2.8t/ha on temporary, permanent and rough-grazing grassland, respectively, accounted for productivity gains by 2010. By 2050, productivities under medium emission scenario are predicted to increase to 15.5 and 9.8t/ha on temporary and permanent grassland, respectively, but not on rough grassland. Based on surveyed grassland distributions for Great Britain in 2010 the annual availability of grassland biomass is likely to rise from 64 to 72milliontonnes by 2050. Assuming optimal N application could close existing productivity gaps of ca. 40% a range of management options could deliver additional 21∗106tonnes of biomass available for bioenergy. Scenarios of changes in grassland use intensity demonstrated considerable scope for maintaining or further increasing grassland production and sparing some grassland for the provision of environmental ecosystem services.

Maxillary expansion in an animal model with light, continuous force.

OBJECTIVES: Maxillary constriction is routinely addressed with rapid maxillary expansion (RME). However, the heavy forces delivered by most RME appliances to expand the palate may lead to deleterious effects on the teeth and supporting tissues. The objective of this study was to explore a more physiologic maxillary expansion with light continuous force. MATERIALS AND METHODS: Twenty 6-week-old Sprague-Dawley rats were equally divided into experimental (EXPT) and control (CTRL) groups. A custom-fabricated archwire expansion appliance made from 0.014-inch copper-nickel-titanium wire was activated 5 mm and bonded to the maxillary molar segments of animals in the EXPT group for 21 days. The force applied to each maxillary segment was 5 cN. Microfocus x-ray computed tomography and histological analyses were used to compare the tooth movement and bone morphology in the midpalatal suture and buccal aspect of the alveolar process between the EXPT and CTRL groups. Descriptive statistics (mean ± standard error of the mean) and nonparametric statistical tests were used to compare the outcomes across groups. RESULTS: Compared to the CTRL group, there was a statistically significant increase in buccal tooth movement and expansion of the midpalatal suture in the EXPT group. There was no difference in the bone morphologic parameters between groups. The mineral apposition rate was increased on the buccal surface of the alveolar process in the EXPT group. CONCLUSIONS: Application of light, continuous force resulted in maxillary osseous expansion due to bilateral sutural apposition and buccal drift of the alveolar processes. This animal experiment provides a more physiologic basis for maxillary expansion.

External validation of a collar-mounted triaxial accelerometer for second-by-second monitoring of eight behavioural states in dogs.

Early detection of disease by an animal owner may motivate them to seek early veterinary advice. Presentation before a more advanced clinical manifestation is evident could lead to more effective treatment and thus benefit the animal's health and welfare. Accelerometers are able to detect changes in specific activities or behaviours, thus indicating early signs of possible adverse health events. The objective of this validation study was to determine whether the detection of eight behavioural states: walk, trot, canter/gallop, sleep, static/inactive, eat, drink, and headshake, by an accelerometer device was sufficiently accurate to be useful in a clinical setting. This fully independent external validation estimated the accuracy of a specific triaxial, collar-mounted accelerometer on a second-by second basis in 51 healthy dogs of different breeds, aged between 6 months and 13 years, weighing >10 kg. The overall diagnostic effectiveness was estimated as: % record correctly classified of > 95% in walk, trot, canter/gallop, eat, drink and headshake and >90% in sleep and static/inactive. The positive predictive values ranged from 93-100%, while the negative predictive values ranged from 96-100%, with exception of static/inactive (86%).This was probably because dogs were placed in unfamiliar kennels where they did not exhibit their typical resting behaviour. The device is worn on a collar, making its use feasible for anyone wanting to monitor their dog's behaviour. The high accuracy in detecting various kinds of behaviour appears promising in assessing canine health and welfare states.

Using Small Angle X-Ray Scattering (SAXS) to Characterize the Solution Conformation and Flexibility of Matrix Metalloproteinases (MMPs).

Small angle X-ray scattering (SAXS) provides information about the conformation and flexibility of proteins in solution, and hence provides complementary structural information to that obtained from X-ray crystallography and nuclear magnetic resonance spectroscopy. In this chapter, we describe the methods for the preparation of matrix metalloproteinase (MMP) samples for SAXS analyses, and for the acquisition, processing and interpretation of the SAXS data.

Global evidence of positive impacts of freshwater biodiversity on fishery yields.

AIM: An often-invoked benefit of high biodiversity is the provision of ecosystem services. However, evidence for this is largely based on data from small-scale experimental studies of relationships between biodiversity and ecosystem function that may have little relevance to real-world systems. Here, large-scale biodiversity datasets are used to test the relationship between the yield of inland capture fisheries and species richness from 100 countries. LOCATION: Inland waters of Africa, Europe and parts of Asia. METHODS: A multimodel inference approach was used to assess inland fishery yields at the country level against species richness, waterside human population, area, elevation and various climatic variables, to determine the relative importance of species richness to fisheries yields compared with other major large-scale drivers. Secondly, the mean decadal variation in fishery yields at the country level for 1981-2010 was regressed against species richness to assess if greater diversity reduces the variability in yields over time. RESULTS: Despite a widespread reliance on targeting just a few species of fish, freshwater fish species richness is highly correlated with yield (R2 = 0.55) and remains an important and statistically significant predictor of yield once other macroecological drivers are controlled for. Freshwater richness also has a significant negative relationship with variability of yield over time in Africa (R2 = 0.16) but no effect in Europe. MAIN CONCLUSIONS: The management of inland waters should incorporate the protection of freshwater biodiversity, particularly in countries with the highest-yielding inland fisheries as these also tend to have high freshwater biodiversity. As these results suggest a link between biodiversity and stable, high-yielding fisheries, an important win-win outcome may be possible for food security and conservation of freshwater ecosystems. However, findings also highlight the urgent need for more data to fully understand and monitor the contribution of biodiversity to inland fisheries globally.