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Background: Access to surgical specialty care differs based on geographic location, insurance status, and subspecialty type. This study uses the Inland Empire as a model to determine the relationship between Social Vulnerability Indices (SVIs), surgeon sex, and surgical subspecialty distribution. Methods: 823 census tracts from the Centers for Disease Control's (CDC) SVI 2018 database were compared against 992 surgeons within 30 distinct subspecialties. This data was retrieved from the American Medical Association's (AMA) 2018 Physician Masterfile. Spearman's bivariate and multiple regression were used to compare the relationship between SVI and number of surgical subspecialists within each census tract. Results: There were approximately 3.34 male and 0.35 female surgeons per census tract (t(267) = 7.74, p

Lipid nanoparticles (LNPs) are being intensively researched and developed to leverage their ability to safely and effectively deliver therapeutics. To achieve optimal therapeutic delivery, a comprehensive understanding of the relationship between formulation, structure, and efficacy is critical. However, the vast chemical space involved in the production of LNPs and the resulting structural complexity make the structure to function relationship challenging to assess and predict. New components and formulation procedures, which provide new opportunities for the use of LNPs, would be best identified and optimized using high-throughput characterization methods. Recently, a high-throughput workflow, consisting of automated mixing, small-angle X-ray scattering (SAXS), and cellular assays, demonstrated a link between formulation, internal structure, and efficacy for a library of LNPs. As SAXS data can be rapidly collected, the stage is set for the collection of thousands of SAXS profiles from a myriad of LNP formulations. In addition, correlated LNP small-angle neutron scattering (SANS) datasets, where components are systematically deuterated for additional contrast inside, provide complementary structural information. The centralization of SAXS and SANS datasets from LNPs, with appropriate, standardized metadata describing formulation parameters, into a data repository will provide valuable guidance for the formulation of LNPs with desired properties. To this end, we introduce Simple Scattering, an easy-to-use, open data repository for storing and sharing groups of correlated scattering profiles obtained from LNP screening experiments. Here, we discuss the current state of the repository, including limitations and upcoming changes, and our vision towards future usage in developing our collective knowledge base of LNPs.

Abstract A culturally tailored virtual program could meet the survivorship needs of Asian American women breast cancer survivors (AABC). This study aims to determine the efficacy of a culturally tailored virtual information and coaching/support program (TICAA) in improving AABC’s survivorship experience. A randomized clinical trial (NCT02803593) was conducted from January 2017 to June 2020 among 199 AABC. The intervention group utilized TICAA and the American Cancer Society [ACS] website while the control group used only ACS website for 12 weeks. The outcomes were measured using the SCNS-34SF (needs; primary), the MSAS-SF (symptoms; secondary), and the FACT-B (quality of life; secondary). The data were analyzed using an intent-to-treat approach. The intervention group showed significant reductions in their needs from the baseline (T0) to post 4 weeks (T1) and to post 12 weeks (T2). Although the changes were not statistically significant, the intervention group had decreased symptoms from T0 to T2 while the control group had an increase in their symptoms. The intervention group had a significant increase in their quality of life from T0 to T2. A culturally tailored virtual program could therefore improve quality of life in AABC patients. Trial Registration: To Enhance Breast Cancer Survivorship of Asian Americans (TICAA), NCT02803593, https://clinicaltrials.gov/ct2/show/NCT02803593?titles=TICAA&draw=2&rank=1

Abstract. Objectives:. With more than 300,000 patients per year in the United States alone, hip fractures are one of the most common injuries occurring in the elderly. The incidence is predicted to rise to 6 million cases per annum worldwide by 2050. Many fracture registries have been established, serving as tools for quality surveillance and evaluating patient outcomes. Most registries are based on billing and procedural codes, prone to under-reporting of cases. Deep learning (DL) is able to interpret radiographic images and assist in fracture detection; we propose to conduct a DL-based approach intended to autocreate a fracture registry, specifically for the hip fracture population. Methods:. Conventional radiographs (n = 18,834) from 2919 patients from Massachusetts General Brigham hospitals were extracted (images designated as hip radiographs within the medical record). We designed a cascade model consisting of 3 submodules for image view classification (MI), postoperative implant detection (MII), and proximal femoral fracture detection (MIII), including data augmentation and scaling, and convolutional neural networks for model development. An ensemble model of 10 models (based on ResNet, VGG, DenseNet, and EfficientNet architectures) was created to detect the presence of a fracture. Results:. The accuracy of the developed submodules reached 92%–100%; visual explanations of model predictions were generated through gradient-based methods. Time for the automated model-based fracture–labeling was 0.03 seconds/image, compared with an average of 12 seconds/image for human annotation as calculated in our preprocessing stages. Conclusion:. This semisupervised DL approach labeled hip fractures with high accuracy. This mitigates the burden of annotations in a large data set, which is time-consuming and prone to under-reporting. The DL approach may prove beneficial for future efforts to autocreate construct registries that outperform current diagnosis and procedural codes. Clinicians and researchers can use the developed DL approach for quality improvement, diagnostic and prognostic research purposes, and building clinical decision support tools.

Rubella virus is an important human pathogen that can cause neurological deficits in a developing fetus when contracted during pregnancy. Despite successful vaccination programs in the Americas and many developed countries, rubella remains endemic in many regions worldwide and outbreaks occur wherever population immunity is insufficient. Intense interest since rubella virus was first isolated in 1962 has advanced our understanding of clinical outcomes after infection disrupts key processes of fetal neurodevelopment. Yet it is still largely unknown which cell types in the developing brain are targeted. We show that in human brain slices, rubella virus predominantly infects microglia. This infection occurs in a heterogeneous population but not in a highly microglia-enriched monoculture in the absence of other cell types. By using an organoid-microglia model, we further demonstrate that rubella virus infection leads to a profound interferon response in non-microglial cells, including neurons and neural progenitor cells, and this response is attenuated by the presence of microglia.

Background Glioma-induced immune dysregulation of the hematopoietic system has been described in a limited number of studies. In this study, our group further demonstrates that gliomas interrupt the cellular differentiation programming and outcomes of hematopoietic stem and progenitor cells (HSPCs) in the bone marrow. HSPCs from glioma-bearing mice are reprogrammed and driven towards expansion of myeloid lineage precursors and myeloid-derived suppressor cells (MDSCs) in secondary lymphoid organs. However, we found this expansion is reversed by immunotherapy. Adoptive cellular therapy (ACT) has been demonstrably efficacious in multiple preclinical models of central nervous system (CNS) malignancies, and here we describe how glioma-induced dysfunction is reversed by this immunotherapeutic platform.Methods The impact of orthotopic KR158B-luc glioma on HSPCs was evaluated in an unbiased fashion using single cell RNAseq (scRNAseq) of lineage− cells and phenotypically using flow cytometry. Mature myeloid cell frequencies and function were also evaluated using flow cytometry. Finally, ACT containing total body irradiation, tumor RNA-pulsed dendritic cells, tumor-reactive T cells and HSPCs isolated from glioma-bearing or non-tumor-bearing mice were used to evaluate cell fate differentiation and survival.Results Using scRNAseq, we observed an altered HSPC landscape in glioma-bearing versus non-tumor-bearing mice . In addition, an expansion of myeloid lineage subsets, including granulocyte macrophage precursors (GMPs) and MDSCs, were observed in glioma-bearing mice relative to non-tumor-bearing controls. Furthermore, MDSCs from glioma-bearing mice demonstrated increased suppressive capacity toward tumor-specific T cells as compared with MDSCs from non-tumor-bearing hosts. Interestingly, treatment with ACT overcame these suppressive properties. When HSPCs from glioma-bearing mice were transferred in the context of ACT, we observed significant survival benefit and long-term cures in orthotopic glioma models compared with mice treated with ACT using non-glioma-bearing HSPCs.

Objective: Obesity and nutrient oversupply increase mammalian target of rapamycin (mTOR) signaling in multiple cell types and organs, contributing to the onset of insulin resistance and complications of metabolic disease. However, it remains unclear when and where mTOR activation mediates these effects, limiting options for therapeutic intervention. The objective of this study was to isolate the role of constitutive mTOR activation in Nav1.8-expressing peripheral neurons in the onset of diet-induced obesity, bone loss, and metabolic disease. Methods: In humans, loss of function mutations in tuberous sclerosis complex 2 (TSC2) lead to maximal constitutive activation of mTOR. To mirror this in mice, we bred Nav1.8-Cre with TSC2fl/fl animals to conditionally delete TSC2 in Nav1.8-expressing neurons. Male and female mice were studied from 4- to 34-weeks of age and a subset of animals were fed a high-fat diet (HFD) for 24-weeks. Assays of metabolism, body composition, bone morphology, and behavior were performed. Results: By lineage tracing, Nav1.8-Cre targeted peripheral sensory neurons, a subpopulation of postganglionic sympathetics, and several regions of the brain. Conditional knockout of TSC2 in Nav1.8-expressing neurons (Nav1.8-TSC2KO) selectively upregulated neuronal mTORC1 signaling. Male, but not female, Nav1.8-TSC2KO mice had a 4–10% decrease in body size at baseline. When challenged with HFD, both male and female Nav1.8-TSC2KO mice resisted diet-induced gains in body mass. However, this did not protect against HFD-induced metabolic dysfunction and bone loss. In addition, despite not gaining weight, Nav1.8-TSC2KO mice fed HFD still developed high body fat, a unique phenotype previously referred to as ‘normal weight obesity’. Nav1.8-TSC2KO mice also had signs of chronic itch, mild increases in anxiety-like behavior, and sex-specific alterations in HFD-induced fat distribution that led to enhanced visceral obesity in males and preferential deposition of subcutaneous fat in females. Conclusions: Knockout of TSC2 in Nav1.8+ neurons increases itch- and anxiety-like behaviors and substantially modifies fat storage and metabolic responses to HFD. Though this prevents HFD-induced weight gain, it masks depot-specific fat expansion and persistent detrimental effects on metabolic health and peripheral organs such as bone, mimicking the ‘normal weight obesity’ phenotype that is of growing concern. This supports a mechanism by which increased neuronal mTOR signaling can predispose to altered adipose tissue distribution, adipose tissue expansion, impaired peripheral metabolism, and detrimental changes to skeletal health with HFD – despite resistance to weight gain.

Objectives The “marionette technique” for transumbilical laparoscopic cholecystectomy (m-TLC) offers improved cosmesis and possibly shorter postoperative recovery for patient undergoing laparoscopic cholecystectomy versus the four-port conventional laparoscopic cholecystectomy (CLC). We compared the outcomes of m-TLC and CLC at a tertiary care facility in New York. Methods A retrospective chart review was conducted and data on patients who underwent m-TLC and CLC were retrieved. Hospital length of stay (LOS), operative time, and complications were compared between the two groups using linear and logistic regression, as appropriate. Results M-TLC group patients were significantly younger, predominantly females with lower body mass index. They were less likely to have previous abdominal surgery and more likely to have noninflammatory pathology (p

Category: Ankle; Other Introduction/Purpose: Finding factors that can exacerbate or ameliorate the incidence of Venous thromboembolism (VTE) can affect the process of making decision on whether to start prophylaxis or not, especially when on the verge of whether to-give or not-to-give prophylaxis. Among each patient's profile, medications are of the most important factors influencing surgeon's decision on the prophylactic methods in VTE-vulnerable patients. Among medications, Statins were shown to reduce the incidence of VTE in patients who were receiving them for hyperlipidemia and cardiovascular conditions. However, none of the current VTE prediction methods, particularly in orthopaedic practice, have considered statins protective. Herein we aimed to determine any correlations between statin consumption and the incidence of VTE in ankle fractures and whether to include statins in prediction models of VTE. Methods: In this case-control machine learning-based study, approved by the Institutional Review Board (IRB), the ICD and CPT codes were used to identify the patients who were diagnosed with ankle fracture in the Mass General Brigham database from 2004 to end of May 2021. After screening approximately 16,421 patients with ankle fractures, a total of 1,176 patients who were suspect VTE according to their signs and symptoms were recruited, 239 had confirmed VTE (case group) and 937 did not have VTE (controls). Forty-nine cases and 396 controls were statin users. Using a semi-automated machine learning-based algorithm, patients' demographics, past medical and surgical history, fracture characteristics and weber classification, and statin consumption status were obtained, and values were organized in a numerical analyzable manner in the dataset. We used chi-squared and Pearson correlation tests where applicable, and outcomes were displayed and interpreted using p-value (p

Category: Ankle; Other Introduction/Purpose: The risk of venous thromboembolism (VTE) after foot and ankle surgery is significantly lower than rates after hip/knee arthroplasty, but it isn't zero. Specific subgroups of patients may be at higher risk, forcing patients and clinicians to navigate the risks and benefits of chemoprophylaxis with insufficient data. Efforts have been made to add clarity to such decision making using conventional data-analysis and risk-scoring methods, but none of these methods were patient-specific or built on robust models of a given patient's individual characteristics. In this study we used machine-learning to determine the potential risk factors for VTE after ankle fracture. We aimed to develop a patient-specific predictive model to assist clinicians and patients in deciding upon the use of postoperative chemoprophylaxis after foot and ankle surgery. Methods: In this preliminary machine-learning-based case-control study, 16,421 patients with ankle fractures were recruited retrospectively. We used an automated-string search method to find patients who were clinically suspected to have developed VTE. Among 1176 such patients, 239 had confirmed VTE within 180 days of sustaining an ankle fracture (cases) and 937 did not (controls). Groups were further sub-divided in patients who had been receiving chemoprophylaxis and those who hadn't. Over 110 factors and variables including patient demographics, past-medical and surgical history, fracture characteristics, treatment, medications, and laboratory values were included in our machine-learning dataset. Three analytical algorithms were used in our machine-learning methods including backward-logistic-regression, decision-tree-classifier (depth=5), and neural networks (two dense layers (n=16 and 4), two drop-out layers, and a sigmoid classifier). Conventional statistics were also used to compare the case and control groups (chi-squared, t-test, p

Background and purpose — Patient-acceptable symptom states (PASS) represent the level on a patient-reported outcome measure (PROM) at which patients are satisfied with postoperative outcomes. We defined the PASS for the Oxford Hip Score (OHS) and Forgotten Joint Score (FJS-12) at 3-month, 1-year, and 2-year intervals after primary total hip arthroplasty (THA). Patients and methods — Between July 2018 and April 2019, primary THA patients in an academic medical center’s registry completed the OHS, FJS-12, and a satisfaction anchor question at 3-month (n = 230), 1-year (n = 180), or 2-year (n = 187) postoperative intervals. PASS thresholds were derived with receiver operating characteristic analysis using the 80% specificity method. 95% confidence intervals (CI) were calculated using 1,000 non-parametric bootstrap replications. Results — 74%, 85%, and 86% of patients reported having a satisfactory symptom state at 3 months, 1, and 2 years after surgery, respectively. At 3-month, 1-year, and 2-year intervals, PASS thresholds were 34 (CI 31–36), 40 (CI 36–44), and 39 (CI 35–42) points for the OHS and 59 (CI 54–64), 68 (CI 61–75), and 69 (CI 62–75) points for the FJS-12. Interpretation — PASS thresholds varied with time for both the OHS and the FJS-12, with lower 3-month compared with 1-year and 2-year thresholds. These PASS thresholds represent OHS and FJS-12 levels at which the average patient is satisfied with THA outcomes, helping to interpret PROMs and serving as clinically significant benchmarks and patient-centered outcomes for research.

Objective To establish the technique of intratumoral combination therapy of radiofrequency hyperthermia (RFH) with herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) gene therapy for rat ovarian cancers. Material and methods This study consisted of three parts: (1) in vitro experiments to establish the ‘proof of principal’ that combination of RFH and HSV-TK gene therapy has the synergistic effect on human ovarian cancer cells; (2) creation of bioluminescence imaging-detectable rat ovarian cancer model; and (3) in vivo experiments using this rat model to validate the technical feasibility of the combination therapy. Cells and nude rats were divided into four groups: (i) combination therapy (HSV-TK/GCV + RFH); (ii) RFH; (iii) HSV-TK/GCV; and (iv) phosphate-buffered saline (PBS). Data were analyzed using Dunnett t-test or Kruskal–Wallis test. Results Cell proliferation assay demonstrated significantly greater reduction in viable cells with the combination therapy [0.52 (0.43, 0.61)] compared to other treatments [RFH 0.90 (0.84, 0.96), HSV-TK/GCV 0.71 (0.53, 0.88), PBS 1 (1, 1); p

Introduction: For the estimated 15 million cancer survivors in the United States, access to accurate information regarding quality of life issues such as sexual health and fertility preservation is a very important part of cancer recovery. Aim: Because National Cancer Institute (NCI)–designated cancer centers (CCs) are leaders of cancer care, we sought to determine the prevalence and quality of information relating to male and female sexual health and fertility on all NCI-designated CC websites. Methods: A total of 70 NCI-designated CC websites listed on www.cancer.gov and divided by geographic regions were carefully analyzed for the presence and quality of male and female sexual health and fertility information. Fisher’s exact test was performed to assess for differences between websites. Main Outcome Measure: We determined the number and percentage of NCI-designated CC websites with sexual health and/or fertility information, availability of information by region, and information available for each gender. Results: Sexual health and/or fertility information was accessible on 43 of 63 (68.3%) CC websites and was more prevalent on comprehensive CC compared with non-comprehensive CC websites (37 of 49 [75.5%] vs 6 of 14 [42.9%] websites, respectively; P < .05). NCI-designated CC websites had less information on sexual health compared with that on fertility (24 of 63 [38.1%] vs 41 of 63 [65.1%] websites, respectively; P < .01). Access to data was less available for men compared with women (35 of 41 [85.4%] vs 41 of 41 [100%] websites, respectively; P

Abstract Background While prostate cancer can often manifest as an indolent disease, the development of locally-advanced or metastatic disease can cause significant morbidity or mortality. Elucidation of molecular mechanisms contributing to disease progression is crucial for more accurate prognostication and effective treatments. R-Spondin 3 (RSPO3) is a protein previously implicated in the progression of colorectal and lung cancers. However, a role for RSPO3 in prostate cancer prognosis and behaviour has not been explored. Methods We compare the relative levels of RSPO3 expression between normal prostate tissue and prostate cancer in two independent patient cohorts (Taylor and GSE70768—Cambridge). We also examine the association of biochemical relapse with RSPO3 levels in these cohorts. For elucidation of the biological effect of RSPO3, we use siRNA technology to reduce the levels of RSPO3 in established prostate cancer cell lines, and perform in vitro proliferation, invasion, western blotting for EMT markers and clonogenic survival assays for radiation resistance. Furthermore, we show consequences of RSPO3 knockdown in an established chick chorioallantoic membrane (CAM) assay model of metastasis. Results RSPO3 levels are lower in prostate cancer than normal prostate, with a tendency for further loss in metastatic disease. Patients with lower RSPO3 expression have lower rates of biochemical relapse-free survival. SiRNA-mediated loss of RSPO3 results in no change to clonogenic survival and a lower proliferative rate, but increased invasiveness in vitro with induction of epithelial–mesenchymal transition (EMT) markers. Consistent with these results, lower RSPO3 expression translates to greater metastatic capacity in the CAM assay. Together, our preclinical findings identify a role of RSPO3 downregulation in prostate cancer invasiveness, and provide a potential explanation for how RSPO3 functions as a positive prognostic marker in prostate cancer.

We report quantitative label-free imaging with phase and polarization (QLIPP) for simultaneous measurement of density, anisotropy, and orientation of structures in unlabeled live cells and tissue slices. We combine QLIPP with deep neural networks to predict fluorescence images of diverse cell and tissue structures. QLIPP images reveal anatomical regions and axon tract orientation in prenatal human brain tissue sections that are not visible using brightfield imaging. We report a variant of U-Net architecture, multi-channel 2.5D U-Net, for computationally efficient prediction of fluorescence images in three dimensions and over large fields of view. Further, we develop data normalization methods for accurate prediction of myelin distribution over large brain regions. We show that experimental defects in labeling the human tissue can be rescued with quantitative label-free imaging and neural network model. We anticipate that the proposed method will enable new studies of architectural order at spatial scales ranging from organelles to tissue.

X-linked Dystonia-Parkinsonism (XDP) is a progressive neurodegenerative disease involving the loss of medium spiny neurons within the striatum. An XDP-specific haplotype has been identified, consisting of seven sequence variants which cluster around the human TAF1 gene, but a direct relationship between any of these variants and disease pathogenesis has not yet been demonstrated. Because the pathogenic gene lesion remains unclear, it has been difficult to predict cellular pathways which are affected in XDP cells. To address that issue, we assayed expression of defined gene sets in XDP vs. control fibroblasts to identify networks of functionally-related transcripts which may be dysregulated in XDP patient cells. That analysis derived a 51-gene signature distinguishing XDP vs. control fibroblasts which mapped strongly to nuclear factor-kappa B (NFκB), a transcription factor pathway also implicated in the pathogenesis of other neurodegenerative diseases, including Parkinson's (PD) and Huntington's disease (HD). Constitutive and TNFα-evoked NFκB signaling was further evaluated in XDP vs. control fibroblasts based on luciferase reporter activity, DNA binding of NFκB subunits, and endogenous target gene transcription. Compared to control cells, XDP fibroblasts exhibited decreased basal NFκB activity and decreased levels of the active NFκB p50 subunit, but increased target gene expression in response to TNFα. NFκB signaling was further examined in neural stem cells differentiated from XDP and control induced pluripotent stem cell (iPSC) lines, revealing a similar pattern of increased TNFα responses in the patient lines compared to controls. These data indicate that an NFκB signaling phenotype is present in both patient fibroblasts and neural stem cells, suggesting this pathway as a site of dysfunction in XDP.

X-linked dystonia-parkinsonism (XDP) is a hereditary neurodegenerative disorder involving a progressive loss of striatal medium spiny neurons. The mechanisms underlying neurodegeneration are not known, in part because there have been few cellular models available for studying the disease. The XDP haplotype consists of multiple sequence variations in a region of the X chromosome containing TAF1, a large gene with at least 38 exons, and a multiple transcript system (MTS) composed of five unconventional exons. A previous study identified an XDP-specific insertion of a SINE-VNTR-Alu (SVA)-type retrotransposon in intron 32 of TAF1, as well as a neural-specific TAF1 isoform, N-TAF1, which showed decreased expression in post-mortem XDP brain compared with control tissue. Here, we generated XDP patient and control fibroblasts and induced pluripotent stem cells (iPSCs) in order to further probe cellular defects associated with this disease. As initial validation of the model, we compared expression of TAF1 and MTS transcripts in XDP versus control fibroblasts and iPSC-derived neural stem cells (NSCs). Compared with control cells, XDP fibroblasts exhibited decreased expression of TAF1 transcript fragments derived from exons 32-36, a region spanning the SVA insertion site. N-TAF1, which incorporates an alternative exon (exon 34′), was not expressed in fibroblasts, but was detectable in iPSC-differentiated NSCs at levels that were ∼threefold lower in XDP cells than in controls. These results support the previous findings that N-TAF1 expression is impaired in XDP, but additionally indicate that this aberrant transcription might occur in neural cells at relatively early stages of development that precede neurodegeneration.

Serous borderline tumor (SBT) of the testis is a rare, ovarian epithelial-type tumor of the testis. We present a case of SBT after radical orchiectomy in a 59-year-old man who had 3-month progressive enlargement of his right hemi-scrotum, negative testicular tumor markers and scrotal ultrasound equivocal for malignancy. Magnetic resonance imaging (MRI) of the pelvis was obtained to aid with evaluation and showed marked hyperintensity on T2-weighted images with frond-like nodular peripheral enhancement within right testis mass. These distinct MRI findings can help clinicians differentiate SBT from other testis tumors. Relapse has not been observed in 8 years of follow-up.

Molecular characterization of plant cell wall glycosyltransferases is a critical step towards understanding the biosynthesis of the complex plant cell wall, and ultimately for efficient engineering of biofuel and agricultural crops. The majority of these enzymes have proven very difficult to obtain in the needed amount and purity for such molecular studies, and recombinant cell wall glycosyltransferase production efforts have largely failed. A daunting number of strategies can be employed to overcome this challenge, including optimization of DNA and protein sequences, choice of expression organism, expression conditions, co-expression partners, purification methods, and optimization of protein solubility and stability. Hence researchers are presented with thousands of potential conditions to test. Ultimately, the subset of conditions that will be sampled depends on practical considerations and prior knowledge of the enzyme(s) being studied. We have developed a rational approach to this process. We devise a pipeline comprising in silico selection of targets and construct design, and high-throughput expression screening, target enrichment, and hit identification. We have applied this pipeline to a test set of Arabidopsis thaliana cell wall glycosyltransferases known to be challenging to obtain in soluble form, as well as to a library of cell wall glycosyltransferases from other plants including agricultural and biofuel crops. The screening results suggest that recombinant cell wall glycosyltransferases in general have a very low soluble:insoluble ratio in lysates from heterologous expression cultures, and that co-expression of chaperones as well as lysis buffer optimization can increase this ratio. We have applied the identified preferred conditions to Reversibly Glycosylated Polypeptide 1 from Arabidopsis thaliana, and processed this enzyme to near-purity in unprecedented milligram amounts. The obtained preparation of Reversibly Glycosylated Polypeptide 1 has the expected arabinopyranose mutase and autoglycosylation activities.

The structure of ortho-vanillin, C8H8O3, has been revisited with modern methods and at low temperature (100 K). The previous structure [Iwasaki et al. (1976). Acta Cryst. B32, 1264–1266] is confirmed, but geometric precision is improved by an order of magnitude. The C atom of the methoxy group lies close to the benzene ring plane, which is the most common geometry for –OMe groups lying ortho to –OH groups on an aromatic ring. The crystal structure displays one intramolecular O—H...O and three weak intermolecular C—H...O hydrogen bonds.

Abstract Genetic rescue—an increase in population fitness following the introduction of new alleles—has been proven to ameliorate inbreeding depression in small, isolated populations, yet is rarely applied as a conservation tool. A lingering question regarding genetic rescue in wildlife conservation is how long beneficial effects persist in admixed populations. Using data collected over 40 years from 1192 endangered Florida panthers (Puma concolor coryi) across nine generations, we show that the experimental genetic rescue implemented in 1995—via the release of eight female pumas from Texas—alleviated morphological, genetic, and demographic correlates of inbreeding depression, subsequently preventing extirpation of the population. We present unequivocal evidence, for the first time in any terrestrial vertebrate, that genetic and phenotypic benefits of genetic rescue remain in this population after five generations of admixture, which helped increase panther abundance (> fivefold) and genetic effective population size (> 20-fold). Additionally, even with extensive admixture, microsatellite allele frequencies in the population continue to support the distinctness of Florida panthers from other North American puma populations, including Texas. Although threats including habitat loss, human-wildlife conflict, and infectious diseases are challenges to many imperiled populations, our results suggest genetic rescue can serve as an effective, multi-generational tool for conservation of small, isolated populations facing extinction from inbreeding.

This research employs institutional characteristics and market-related factors to predict undergraduate students' tuition at 190 private colleges and universities in the USA. Results showed that the strongest correlations among variables for college tuition were reputation ranking and SAT scores. Results of a hierarchical multiple regression revealed that the type of institution, academic reputation ranking, the annual expenditures, geographic region, the existence of professional schools, the size of the faculty and the undergraduate student body, and university presidents' pay and benefits are all significant predictors of college tuition. After controlling all other variables, the unique contribution made by reputation ranking is still a significant predictor of college tuition. Research institutions charged their students more than liberal arts colleges, which, in turn, charged more than doctoral granting I institutions. Implications for parents and students, private colleges and universities, human resource management, and the Matthew effect are discussed. (Contains 2 tables.)