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Background: This article reports on the findings of a workshop held at the joint 5th World Organisation of Family Doctors (WONCA) Africa and 20th National Family Practitioners Conference in Tshwane, South Africa, in 2017. Postgraduate training for family medicine in Africa takes place in the clinical workspace at the bedside or next to the patient in the clinic, district hospital or regional hospital. Direct supervisor observation, exchange of reflection and feedback, and learning conversations between the supervisor and the registrar are central to learning and assessment processes. Objectives: The aim of the workshop was to understand how family medicine registrars (postgraduate trainees in family medicine) in Africa learn in the workplace. Methods: Thirty-five trainers and registrars from nine African countries, the United Kingdom, United States and Sweden participated. South Africa was represented by the universities of Cape Town, Limpopo, Pretoria, Sefako Makgatho, Stellenbosch, Walter Sisulu and Witwatersrand. Results: Six major themes were identified: (1) context is critical, (2) learning style of the registrar and (teaching style) of the supervisor, (3) learning portfolio is utilised, (4) interactions between registrar and supervisor, (5) giving and receiving feedback and (6) the competence of the supervisor. Conclusion: The training of family physicians across Africa shares many common themes. However, there are also big differences among the various countries and even programmes within countries. The way forward would include exploring the local contextual enablers that influence the learning conversations between trainees and their supervisors. Family medicine training institutions and organisations (such as WONCA Africa and the South African Academy of Family Physicians) have a critical role to play in supporting trainees and trainers towards developing local competencies which facilitate learning in the clinical workplace dominated by service delivery pressures.

Introduction: Brazil and South Africa share many sociodemographic and health features that provide many learning opportunities. Brazil’s national health system, the Sistema Único de Saúde (SUS) prioritises primary health care since 1994, the year democracy came to South Africa. Two family physicians from these countries met in Rocinha favela in Rio de Janeiro, a densely populated area where poverty, danger, drugs, tuberculosis and mental illness are the focus of the health system. Maria do Socorro Family Clinic: Central to the SUS are the Family Health Teams, consisting of community health workers, nurses, doctors and allied health workers. This clinic in Rocinha has 11 teams, caring for 2700 people each, all visited monthly, preventing illness and promoting health. Patients with mental illness are cared for in a therapeutic residency, with an onsite psychiatrist, psychologist and social worker. The relationships between the health carers and the clinic and the community are collegial and equal, sharing care. Larger than life photos of patients from the community line the walls. Training: A culture of learning is evident, with 18 family medicine residents, student nurses, a small library and a learning centre at the clinic. Local authorities compensate trainees in family medicine more than traditional specialties. Conclusion: Brazil has made massive progress in providing universal health coverage over the last 20 years. South Africa, with not too dissimilar challenges, is embarking on this road more recently. The lessons learnt at clinic and community level in this inner-city clinic could be very useful for similar settings in South Africa and other countries.

Background: The inappropriate use of emergency centres (ECs) is an expanding problem globally. The high attendance of non-urgent return presentations to ECs is recognised as part of the problem, placing an unnecessary demand on limited staff and resources. Of unscheduled returns 34% of cases had no change to diagnosis or treatment with the conclusion that 80% of re-attendance could be attributed to deficiencies in the initial consultation. This study aimed to evaluate the reasons why patients sought an early second consultation for the same complaint at a hospital EC in South Africa, by exploring the patient’s experience and shortcomings in the first consultation. Method: A qualitative study was conducted using in-depth, semi-structured interviews with 20 purposively selected participants who presented to a rural regional provincial hospital’s EC within 7 days of a prior consultation for the same complaint. Verbatim transcripts were analysed using the framework method. Results: The main reasons for a second consultation were symptom related factors and the need for diagnostic certainty. The major themes around patient experience of the initial consultation were shortcomings in effective evaluation and management of pain, diagnostic uncertainty including poor examination, poor explanation, uncertain access and follow-up and societal encouragement to utilise a hospital EC. Conclusion: Further interventions should explore pain as a presenting symptom of the illness experience, and promote competence in addressing physical and psychological causative factors within a patient-centred approach for all health staff, especially in primary care services.

Background: Access to health services is one of the Batho Pele (‘people first’) values and principles of the South African government since 1997. This necessitated some changes around public service systems, procedures, attitudes and behaviour. The challenges of providing health care to rural geographically spread populations include variations in socio-economic status, transport opportunities, access to appointment information and patient perceptions of costs and benefits of seeking health care. George hospital, situated in a rural area, serves 5000 outpatient visits monthly, with non-attendance rates of up to 40%. Objectives: The aim of this research was to gain a greater understanding of the reasons behind non-attendance of outpatient department clinics to allow locally driven, targeted interventions. Methods: This was a descriptive study. We attempted to phone all patients who missed appointments over a 1-month period (n = 574). Only 20% were contactable with one person declining consent. Twenty-nine percent had no telephone number on hospital systems, 7% had incorrect numbers, 2% had died and 42% did not respond to three attempts. Results: The main reasons for non-attendance included unaware of appointment date (16%), out of area (11%), confusion over date (11%), sick or admitted to hospital (10%), family member sick or died (7%), appointment should have been cancelled by clerical staff (6%) and transport (6%). Only 9% chose to miss their appointment. The other 24% had various reasons. Conclusions: Improved patient awareness of appointments, adjustments in referral systems and enabling appointment cancellation if indicated would directly improve over two-thirds of reasons for non-attendance. Understanding the underlying causes will help appointment planning, reduce wasted costs and have a significant impact on patient care.

The Space Shuttle Atlantis launched on its final mission (STS-135) on July 8, 2011. After just under 13 days, the shuttle landed safely at Kennedy Space Center (KSC) for the last time. Female C57BL/6J mice flew as part of the Commercial Biomedical Testing Module-3 (CBTM-3) payload. Ground controls were maintained at the KSC facility. Subsets of these mice were made available to investigators as part of NASA's Bio-specimen Sharing Program (BSP). Our group characterized cell phenotype distributions and phagocytic function in the spleen, catecholamine and corticosterone levels in the adrenal glands, and transcriptomics/metabolomics in the liver. Despite decreases in most splenic leukocyte subsets, there were increases in reactive oxygen species (ROS)-related activity. Although there were increases noted in corticosterone levels in both the adrenals and liver, there were no significant changes in catecholamine levels. Furthermore, functional analysis of gene expression and metabolomic profiles suggest that the functional changes are not due to oxidative or psychological stress. Despite changes in gene expression patterns indicative of increases in phagocytic activity (e.g. endocytosis and formation of peroxisomes), there was no corresponding increase in genes related to ROS metabolism. In contrast, there were increases in expression profiles related to fatty acid oxidation with decreases in glycolysis-related profiles. Given the clear link between immune function and metabolism in many ground-based diseases, we propose a similar link may be involved in spaceflight-induced decrements in immune and metabolic function.

Protein diagnostic applications typically require pairs of analyte-specific reagents for capture and detection. We developed methods for the systematic isolation of slow off-rate modified aptamer (SOMAmer) reagents that bind to different epitopes and allow efficient pair-wise screening of multiple ligands. SOMAmers were generated via a second systematic evolution of ligands by exponential enrichment (SELEX), using complexes of target proteins with a primary, non-amplifiable SOMAmer and employing different modified nucleotides (e.g., naphthylmethyl- or tryptaminocarbonyl-dU) to favor alternate binding epitopes. Non-competing binding of primary and secondary SOMAmers was tested in radiolabel competition and sandwich binding assays. Multiplexed high-throughput screening for sandwich pairs utilized the Luminex platform, with primary SOMAmers as capture agents attached to different types of LumAvidin beads, which were then pooled for testing the secondary SOMAmers individually as detection agents. Functional SOMAmer pairs were obtained for Clostridium difficile binary toxin (CdtA) and for a panel of human proteins (ANGPT2, TSP2, CRDL1, MATN2, GPVI, C7, PLG) that had been previously identified as promising markers for cardiovascular risk. The equilibrium dissociation constants (Kd values) ranged from 0.02–2.7 nM, and the detection limits were in the low picomolar range for these proteins in SOMAmer sandwich assays. These results indicate that SOMAmer pairs hold promise for the development of rapid tests or specific diagnostic panels.

We measured the gas exchange parameters of six onion varieties in two photoperiodic regimes. The photoperiods simulated early (long-day) and late (short-day) onion growing seasons of the high desert regions near Lancaster, California. We used six locally grown onion varieties: RedBull (RB), RedWing (RW), Hybrid Cometa (HC), Granero (YG), Hybrid Valero (HV) and Hybrid SR (HS). For both photoperiod regimes, daylength and temperature were changed weekly to represent the natural progression of field growing conditions. The results show that the onion plants grown under short-daylength (SD) and higher temperature regimes exhibited higher leaf photosynthetic rates and stomatal conductance than onion seedlings grown under the long-day (LD) regime. Onion plants also had different functional leaf traits depending on crop growth environment during the onset of bulbing. The higher photosynthetic rate of the short-day plants coincided with bulb initiation. Onion photosynthesis and stomatal conductance increased at bulb initiation, apparently to provide carbohydrates for growth and storage in the bulb.

Building on the research about assessing teachers' leader identity, we were curious to understand how K-12 school principals perceive teachers as leaders. We used a survey research design to gather quantitative and qualitative data from K-12 school principals. We could not find any extant surveys aligned with our research focus; thus, we designed and validated a survey for our research. We contacted 1042 K-12 principals from the south-central region of the US; 70 responded by completing our survey. Analysis indicated a potential misalignment in principal leadership styles between the ideal attributes of leadership and contextual or practical applications of leadership. Additional analysis revealed considerable overlap between the principals' perceptions of leadership and the roles of teachers; however, those leadership roles aligned with teachers as school-level leaders outside the classroom rather than in their role of teaching students. Findings also revealed an alignment of principals' perceptions of teachers as leaders with principals' efforts to develop teachers as leaders. Overall, our research suggests that principals support teachers' development as leaders. We close with a discussion of the results and implications for research and practice.

We explored the challenges with the unplanned change of abruptly shifting to online learning that the COVID-19 pandemic mitigation had on first-year students' perceptions of learning, their connection to the university, and the general college experience. We used a cross-sectional method to gather quantitative and qualitative data using an online survey. More than 200 participating first-year college students indicated concerns with making connections, feeling challenged and unfulfilled with their educational experience, and struggling to adjust to the pandemic-mandated changes. Additionally, we found that students experienced challenges accepting ownership of their learning and navigating ambiguous situations.

Reading and writing are fundamental skills students need to succeed in college, making literacy development an issue of education equity. The literacy skills can be content-specific, indicating faculty members across disciplines need to support student development of appropriate literacy skills. The extent to which faculty members support student literacy development is likely associated with their literacy-focused education equity mindset. The goal of our research was to document the mindset of faculty members across multiple disciplines. We gathered a combination of quantitative and qualitative data from 345 college faculty members using a survey. We found variations in the mindset strength between disciplines by the number of students taught, gender, and age. Overall, the faculty members held a moderate literacy-focused education equity mindset. The findings have implications for student inclusion, retention, and completion.

Mycobacteria produce a capsule layer, which consists of glycan-like polysaccharides and a number of specific proteins. In this study, we show that, in slow-growing mycobacteria, the type VII secretion system ESX-5 plays a major role in the integrity and stability of the capsule. We have identified PPE10 as the ESX-5 substrate responsible for this effect. Mutants in esx-5 and ppe10 both have impaired capsule integrity as well as reduced surface hydrophobicity. Electron microscopy, immunoblot and flow cytometry analyses demonstrated reduced amounts of surface localized proteins and glycolipids, and morphological differences in the capsular layer. Since capsular proteins secreted by the ESX-1 system are important virulence factors, we tested the effect of the mutations that cause capsular defects on virulence mechanisms. Both esx-5 and ppe10 mutants of Mycobacterium marinum were shown to be impaired in ESX-1-dependent hemolysis. In agreement with this, the ppe10 and esx5 mutants showed reduced recruitment of ubiquitin in early macrophage infection and intermediate attenuation in zebrafish embryos. These results provide a pivotal role for the ESX-5 secretion system and its substrate PPE10, in the capsular integrity of pathogenic mycobacteria. These findings open up new roads for research on the mycobacterial capsule and its role in virulence and immune modulation.

Background: Since 2007, the postgraduate training of family physicians for South African district hospitals has been formalised. This training differs from European and North American programmes as up to 30% of the skills needed rely on district hospital surgical, obstetrics and anaesthetics procedures, particularly in rural areas, as outlined in the national unit standards. The aim of this study was to evaluate the appropriateness and sufficiency of learning opportunities for these skills in a rural district hospital. Methods: A descriptive, cross-sectional study was undertaken of the number and type of procedures performed in theatre for a 1-year period and compared with the required procedural skills stipulated in the national unit standards. Descriptive statistical analyses were used to analyse categorical data. Results: Three thousand seven hundred and forty-one procedures were performed during the study period. Anaesthesia was the most common procedure, followed by Caesarean section. There were adequate opportunities for teaching most core skills. Conclusions: Sufficient and appropriate learning opportunities exist for postgraduate family medicine training in all the core skills performed in a theatre according to the national unit standards. Keywords: Post Graduate Training, Family Medicine, Procedural Skills, Rural, District hospitals

Introduction: Radical cystectomy (RC) is the gold standard treatment for muscle-invasive bladder cancer. This procedure has a high rate of perioperative complications, many of which are infectious in nature. The objective of our study was to evaluate demographic, intrinsic and extrinsic patient variables associated with developing readmission within 30 days due to infectious complications following RC. Methods: We acquired data available from the American College of Surgeons National Surgical Quality Improvement Program. We queried this dataset to identify all patients who underwent RC for muscle-invasive malignant disease (CPT 188.x) in 2012 based on CPT coding. Logistic regression analysis was used to investigate the relationship between preoperative variables and readmissions for infectious complications. Results: Of the 961 patients undergoing cystectomy for malignancy, 159 (17%) required readmission for any indications at a median of 16 days (interquartile range 13–22 days) postoperatively. We identified 71 of a total of 159 (45%) readmissions, which were due to infectious complications. Smoking was more prevalent in the patient population readmitted for an infectious complication compared with the patient population readmitted for a non-infectious complication (37% versus 25%; p = 0.03). Using logistic regression analysis smoking was associated with a significant risk for readmission due to an infectious cause (odds ratio 2.28, 95% confidence interval 1.82–2.97, p = 0.02). Readmission due to an infectious etiology was not associated with other perioperative factors including type of urinary diversion, sex, duration of operation, hypertension, or recent weight loss. Conclusion: Readmission following RC is a common occurrence and infectious complications drive readmission in almost half of the cases. Current smoking was the only independent risk factor for an infectious readmission. Counseling patients in smoking cessation prior to the procedure may provide an avenue for quality improvement to limit readmissions.

Volatile chemicals increased trap catch of flies from the families Lauxaniidae [Homoneura bispina (Loew) and Camptoprosopella borealis Shewell], Chloropidae (Olcella sp.), and Anthomyiidae (Delia spp.) in field crops. With lauxaniids, baiting with 2-phenylethanol on cotton-roll dispensers increased catch of H. bispina in two corn plot tests, and methyl salicylate increased trap catch in one test. Traps baited with methyl salicylate increased the catch of C. borealis. When using plastic-sachet dispensers, traps baited with methyl salicylate caught more H. bispina than ones baited with 2-phenylethanol, whereas traps baited with 2-phenylethanol caught more C. borealis than those with methyl salicylate. For chloropids, traps baited with 2-isopropyl-3-methoxypyrazine greatly increased catch of Olcella flies in corn and soybean. With anthomyiids, catch of male Delia flies in wheat increased with 2-phenylethanol on cotton rolls and with either 2-phenylethanol or methyl salicylate using plastic dispensers. In soybean, 2-phenylethanol formulated on cotton rolls or in plastic dispensers increased catch of male Delia flies, but methyl salicylate did not affect trap catch. Trap catch of female Delia flies did not vary among chemicals. In another test in soybean, trap catch of both male and female Delia flies was greater with 2-phenylethanol than with other volatile chemicals.

Spaceflight affects numerous organ systems in the body, leading to metabolic dysfunction that may have long-term consequences. Microgravity-induced alterations in liver metabolism, particularly with respect to lipids, remain largely unexplored. Here we utilize a novel systems biology approach, combining metabolomics and transcriptomics with advanced Raman microscopy, to investigate altered hepatic lipid metabolism in mice following short duration spaceflight. Mice flown aboard Space Transportation System -135, the last Shuttle mission, lose weight but redistribute lipids, particularly to the liver. Intriguingly, spaceflight mice lose retinol from lipid droplets. Both mRNA and metabolite changes suggest the retinol loss is linked to activation of PPARα-mediated pathways and potentially to hepatic stellate cell activation, both of which may be coincident with increased bile acids and early signs of liver injury. Although the 13-day flight duration is too short for frank fibrosis to develop, the retinol loss plus changes in markers of extracellular matrix remodeling raise the concern that longer duration exposure to the space environment may result in progressive liver damage, increasing the risk for nonalcoholic fatty liver disease.

[This corrects the article DOI: 10.1371/journal.pone.0152877.].

Kate Van Brunt1, Louis S Matza1, Peter M Classi2, Joseph A Johnston21Center for Health Outcomes Research at United BioSource Corporation, Bethesda, MD, USA; 2Eli Lilly and Company, Indianapolis, IN, USAObjectives: A growing body of literature has highlighted the importance of considering patient preferences as part of the medical decision-making process. The purpose of the current review was to identify and summarize published research on preferences related to attention-deficit/hyperactivity disorder (ADHD) and its treatment, while suggesting directions for future research.Methods: A literature search identified 15 articles that included a choice-based assessment of preferences related to ADHD.Results: The 15 studies were grouped into four categories based on preference content: preference for a treatment directly experienced by the respondent or the respondent's child; preference for general treatment approaches; preference for a specific treatment attribute or outcome; and preference for aspects of ADHD-related treatment. Preference assessment methods ranged from global single items to detailed choice-based procedures, with few studies using rigorously developed assessment methods. Respondents included patients with ADHD, clinicians, parents, teachers, and survey respondents from the general population. Factors influencing preference include treatment characteristics, effectiveness for specific symptoms, side effects, and respondent demographics. Minimal research has examined treatment preferences of adults with ADHD.Discussion: Because there is no dominant treatment known to be the first choice for all patients, ADHD is a condition for which individual preferences can play an important role when making treatment decisions for individual patients. Given the potential role of preferences in clinical decision-making, more research is needed to better understand the preferences of patients with ADHD and other individuals who are directly affected by the disorder, such as parents and teachers.Keywords: patient preference, ADHD, parent preference, utility

PurposeThe Audio-Visual Assisted Therapeutic Ambience in Radiotherapy (AVATAR) system was the first published radiation therapy (RT)-compatible system to reduce the need for pediatric anesthesia through video-based distraction. We evaluated the feasibility of AVATAR implementation and effects on anesthesia use, quality of life, and anxiety in a multicenter pediatric trial.Methods and materialsPediatric patients 3 to 10 years of age preparing to undergo RT at 10 institutions were prospectively enrolled. Children able to undergo at least 1 fraction of RT using AVATAR without anesthesia were considered successful (S). Patients requiring anesthesia for their entire treatment course were nonsuccessful (NS). The PedsQL3.0 Cancer Module (PedsQL) survey assessed quality of life and was administered to the patient and guardian at RT simulation, midway through RT, and at final treatment. The modified Yale Preoperative Anxiety Scale (mYPAS) assessed anxiety and was performed at the same 3 time points. Success was evaluated using the χ2 test. PedsQL and mYPAS scores were assessed using mixed effects models with time points evaluated as fixed effects and a random intercept on the subject.ResultsEighty-one children were included; median age was 7 years. AVATAR was successful at all 10 institutions and with photon and proton RT. There were 63 (78%) S patients; anesthesia was avoided for a median of 20 fractions per patient. Success differed by age (P = .04) and private versus public insurance (P < .001). Both patient (P = .008) and parent (P = .006) PedsQL scores significantly improved over the course of RT for patients aged 5 to 7. Anxiety in the treatment room decreased for both S and NS patients over RT course (P < .001), by age (P < .001), and by S versus NS patients (P < .001).ConclusionsIn this 10-center prospective trial, anesthesia avoidance with AVATAR was 78% in children aged 3 to 10 years, higher than among age-matched historical controls (49%; P < .001). AVATAR implementation is feasible across multiple institutions and should be further studied and made available to patients who may benefit from video-based distraction.

Microarray gene expression profiling has been used to distinguish histological subtypes of renal cell carcinoma (RCC), and consequently to identify specific tumor markers. The analytical procedures currently in use find sets of genes whose average differential expression across the two categories differ significantly. In general each of the markers thus identifi ed does not distinguish tumor from normal with 100% accuracy, although the group as a whole might be able to do so. For the purpose of developing a widely used economically viable diagnostic signature, however, large groups of genes are not likely to be useful. Here we use two different methods, one a support vector machine variant, and the other an exhaustive search, to reanalyze data previously generated in our Lab (Lenburg et al. 2003). We identify 158 genes, each having an expression level that is higher (lower) in every tumor sample than in any normal sample, and each having a minimum differential expression across the two categorie at a signifi cance of 0.01. The set is highly enriched in cancer related genes (p = 1.6 × 10 – 12), containing 43 genes previously associated with either RCC or other types of cancer. Many of the biomarkers appear to be associated with the central alterations known to be required for cancer transformation. These include the oncogenes JAZF1, AXL, ABL2; tumor suppressors RASD1, PTPRO, TFAP2A, CDKN1C; and genes involved in proteolysis or cell-adhesion such as WASF2, and PAPPA.

Background: The theme of the 2014 Southern African Rural Health Conference was ‘Building resilience in facing rural realities’. Retaining health professionals in South Africa is critical for sustainable health services. Only 12% of doctors and 19% of nurses have been retained in the rural areas. The aim of the workshop was to understand from health practitioners why they continued working in their rural settings. Conference workshop: The workshop consisted of 29 doctors, managers, academic family physicians, nurses and clinical associates from Southern Africa, with work experience from three weeks to 13 years, often in deep rural districts. Using the nominal group technique, the following question was explored, ‘What is it that keeps you going to work every day?’ Participants reflected on their work situation and listed and rated the important reasons for continuing to work. Results: Five main themes emerged. A shared purpose, emanating from a deep sense of meaning, was the strongest reason for staying and working in a rural setting. Working in a team was second most important, with teamwork being related to attitudes and relationships, support from visiting specialists and opportunities to implement individual clinical skills. A culture of support was third, followed by opportunities for growth and continuing professional development, including teaching by outreaching specialists. The fifth theme was a healthy work-life balance. Conclusion: Health practitioners continue to work in rural settings for often deeper reasons relating to a sense of meaning, being part of a team that closely relate to each other and feeling supported.

Mycobacteria possess different type VII secretion (T7S) systems to secrete proteins across their unusual cell envelope. One of these systems, ESX-5, is only present in slow-growing mycobacteria and responsible for the secretion of multiple substrates. However, the role of ESX-5 substrates in growth and/or virulence is largely unknown. In this study, we show that esx-5 is essential for growth of both Mycobacterium marinum and Mycobacterium bovis. Remarkably, this essentiality can be rescued by increasing the permeability of the outer membrane, either by altering its lipid composition or by the introduction of the heterologous porin MspA. Mutagenesis of the first nucleotide-binding domain of the membrane ATPase EccC5 prevented both ESX-5-dependent secretion and bacterial growth, but did not affect ESX-5 complex assembly. This suggests that the rescuing effect is not due to pores formed by the ESX-5 membrane complex, but caused by ESX-5 activity. Subsequent proteomic analysis to identify crucial ESX-5 substrates confirmed that all detectable PE and PPE proteins in the cell surface and cell envelope fractions were routed through ESX-5. Additionally, saturated transposon-directed insertion-site sequencing (TraDIS) was applied to both wild-type M. marinum cells and cells expressing mspA to identify genes that are not essential anymore in the presence of MspA. This analysis confirmed the importance of esx-5, but we could not identify essential ESX-5 substrates, indicating that multiple of these substrates are together responsible for the essentiality. Finally, examination of phenotypes on defined carbon sources revealed that an esx-5 mutant is strongly impaired in the uptake and utilization of hydrophobic carbon sources. Based on these data, we propose a model in which the ESX-5 system is responsible for the transport of cell envelope proteins that are required for nutrient uptake. These proteins might in this way compensate for the lack of MspA-like porins in slow-growing mycobacteria.

We evaluated the performance of the Heat Dissipation Technique (HDT) to measure sap flow in whole trees by comparison with potometric water uptake. Two tropical lowland species, Ochroma lagopus (balsa), a pioneer species with light wood and Hyeronima alchorneoides (pilón), a late-successional species with hard wood were examined. Diurnal courses of sap flow measured with the HDT showed good agreement with potometry. At the low sap flow rates (below 1 Kg h-1) occurring during nocturnal recharge HDT consistently underestimated sap flow rates. This resulted in the failure of the current version of the HDT to measure nocturnal water uptake, an important component of the water budget of at least one of the two species examined. Rev. Biol. Trop. 54(1): 73-81. Epub 2006 Mar 31.

A tumour microenvironment abundant in regulatory T (Treg) cells aids solid tumours to evade clearance by effector T cells. Systemic strategies to suppress Treg cells or to augment immunity can elicit autoimmune side effects, cytokine storms and other toxicities. Here we report the design, fabrication and therapeutic performance of a biodegradable macroporous scaffold, implanted peritumourally, that releases a small-molecule inhibitor of transforming growth factor β to suppress Treg cells, chemokines to attract effector T cells and antibodies to stimulate them. In two mouse models of aggressive tumours, the implant boosted the recruitment and activation of effector T cells into the tumour and depleted it of Treg cells, which resulted in an 'immunological abscopal effect' on distant metastases and in the establishment of long-term memory that impeded tumour recurrence. We also show that the scaffold can be used to deliver tumour-antigen-specific T cells into the tumour. Peritumourally implanted immunomodulatory scaffolds may represent a general strategy to enhance T-cell immunity and avoid the toxicities of systemic therapies.

Microgravity-induced bone loss results in a 1% bone mineral density loss monthly and can be a mission critical factor in long-duration spaceflight. Biomolecular therapies with dual osteogenic and anti-resorptive functions are promising for treating extreme osteoporosis. We previously confirmed that NELL-like molecule-1 (NELL-1) is crucial for bone density maintenance. We further PEGylated NELL-1 (NELL-polyethylene glycol, or NELL-PEG) to increase systemic delivery half-life from 5.5 to 15.5 h. In this study, we used a bio-inert bisphosphonate (BP) moiety to chemically engineer NELL-PEG into BP-NELL-PEG and specifically target bone tissues. We found conjugation with BP improved hydroxyapatite (HA) binding and protein stability of NELL-PEG while preserving NELL-1's osteogenicity in vitro. Furthermore, BP-NELL-PEG showed superior in vivo bone specificity without observable pathology in liver, spleen, lungs, brain, heart, muscles, or ovaries of mice. Finally, we tested BP-NELL-PEG through spaceflight exposure onboard the International Space Station (ISS) at maximal animal capacity (n = 40) in a long-term (9 week) osteoporosis therapeutic study and found that BP-NELL-PEG significantly increased bone formation in flight and ground control mice without obvious adverse health effects. Our results highlight BP-NELL-PEG as a promising therapeutic to mitigate extreme bone loss from long-duration microgravity exposure and musculoskeletal degeneration on Earth, especially when resistance training is not possible due to incapacity (e.g., bone fracture, stroke).

Urbanization creates novel ecosystems comprised of species assemblages and environments with no natural analogue. Moreover, irrigation can alter plant function compared to non-irrigated systems. However, the capacity of irrigation to alter functional trait patterns across multiple species is unknown but may be important for the dynamics of urban ecosystems. We evaluated the hypothesis that urban irrigation influences plasticity in functional traits by measuring carbon-gain and water-use traits of 30 tree species planted in Southern California, USA spanning a coastal-to-desert gradient. Tree species respond to irrigation through increasing the carbon-gain trait relationship of leaf nitrogen per specific leaf area compared to their native habitat. Moreover, most species shift to a water-use strategy of greater water loss through stomata when planted in irrigated desert-like environments compared to coastal environments, implying that irrigated species capitalize on increased water availability to cool their leaves in extreme heat and high evaporative demand conditions. Therefore, irrigated urban environments increase the plasticity of trait responses compared to native ecosystems, allowing for novel response to climatic variation. Our results indicate that trees grown in water-resource-rich urban ecosystems can alter their functional traits plasticity beyond those measured in native ecosystems, which can lead to plant trait dynamics with no natural analogue.

BackgroundPatient-reported outcome measures are needed to assess the impact of treatments for COVID-19 on symptoms. The ACTIV-2 COVID-19 Symptom Diary (ACSD) is being used in the ongoing Accelerating COVID-19 Therapeutic Interventions and Vaccines-2 (ACTIV-2) platform clinical trial. The purpose of the current study was to conduct qualitative interviews to assess content validity of the ACSD.MethodsInterviews were conducted with adults who had tested positive for SARS-CoV-2. The ACSD begins with global items, followed by a symptom checklist. Each interview began with concept elicitation focusing on participant experiences with COVID-19. Then, participants completed the ACSD, and cognitive interviews were conducted to evaluate the questionnaire. Interviews were recorded, transcribed, and coded following a qualitative content analysis. For the qualitative analysis, a coding dictionary was developed with a list of all potential codes and instructions for how the codes should be applied and combined.ResultsInterviews were conducted with 30 participants (mean age = 39 years; 57% female; 17% Latinx; 17% Black/African American; 40% meeting at least one criterion for classification as high risk of progression to severe COVID-19). Commonly reported symptoms included fatigue (reported by 100% of the sample), body pain/muscle pain/aches (87%), headaches (87%), cough (83%), loss of smell (73%), shortness of breath/difficulty breathing (70%), and chills (70%). The 13 symptoms most commonly reported in this study are included in the ACSD. After completing the ACSD, participants consistently reported that it was clear and easy to complete, and all items were generally interpreted as intended. Based on participants' input, the ACSD was edited slightly after the first 13 interviews, and the revised version was used for the final 17 interviews. Two additional items assessing "brain fog" and dizziness were recommended for addition to the ACSD in future research.ConclusionsThis qualitative study supports the content validity of the ACSD for assessment of COVID-19 symptoms. Quantitative research with larger samples will be needed to examine the questionnaire's measurement properties.

Urothelial cells respond to bladder distension with ATP release, and ATP signaling within the bladder and from the bladder to the CNS is essential for proper bladder function. In other cell types, pannexin 1 (Panx1) channels provide a pathway for mechanically-induced ATP efflux and for ATP-induced ATP release through interaction with P2X7 receptors (P2X7Rs). We report that Panx1 and P2X7R are functionally expressed in the bladder mucosa and in immortalized human urothelial cells (TRT-HU1), and participate in urothelial ATP release and signaling. ATP release from isolated rat bladders induced by distention was reduced by the Panx1 channel blocker mefloquine (MFQ) and was blunted in mice lacking Panx1 or P2X7R expression. Hypoosmotic shock induced YoPro dye uptake was inhibited by MFQ and the P2X7R blocker A438079 in TRT-HU1 cells, and was also blunted in primary urothelial cells derived from mice lacking Panx1 or P2X7R expression. Rinsing-induced mechanical stimulation of TRT-HU1 cells triggered ATP release, which was reduced by MFQ and potentiated in low divalent cation solution (LDPBS), a condition known to enhance P2X7R activation. ATP signaling evaluated as intercellular Ca2+ wave radius was significantly larger in LDPBS, reduced by MFQ and by apyrase (ATP scavenger). These findings indicate that Panx1 participates in urothelial mechanotransduction and signaling by providing a direct pathway for mechanically-induced ATP release and by functionally interacting with P2X7Rs.

The identification of new drug candidates from chemical libraries is a major component of discovery research in many pharmaceutical companies. Given the large size of many conventional and combinatorial libraries and the rapid increase in the number of possible therapeutic targets, the speed with which efficient high-throughput screening (HTS) assays can be developed can be a rate-limiting step in the discovery process. We show here that aptamers, nucleic acids that bind other molecules with high affinity, can be used as versatile reagents in competition binding HTS assays to identify and optimize small-molecule ligands to protein targets. To illustrate this application, we have used labeled aptamers to plateletderived growth factor B-chain and wheat germ agglutinin to screen two sets of potential small-molecule ligands. In both cases, binding affinities of all ligands tested (small molecules and aptamers) were strongly correlated with their inhibitory potencies in functional assays. The major advantages of using aptamers in HTS assays are speed of aptamer identification, high affinity of aptamers for protein targets, relatively large aptamer-protein interaction surfaces, and compatibility with various labeling/detection strategies. Aptamers may be particularly useful in HTS assays with protein targets that have no known binding partners such as orphan receptors. Since aptamers that bind to proteins are often specific and potent antagonists of protein function, the use of aptamers for target validation can be coupled with their subsequent use in HTS.

Substantial amounts of nitrate nitrogen NO3-N can leach from intensively grazed pasture in the northeast U.S. where there is about 30 cm of groundwater recharge, annually. Management options for reducing NO3-N leaching were evaluated for this environment using the Cornell Net Carbohydrate and Protein System Model and a recently developed nitrogen leaching index. Management options utilizing energy supplementation of grazing dairy cows could improve nitrogen efficiency within the cow, but would not necessarily reduce NO3-N leaching at the pasture scale if stocking rate was not controlled. The management option of using white clover to supply nitrogen to the pasture decreased NO3-N leaching, but produced less dry matter yield, which in turn reduced stocking rate. The economic returns of reducing NO3-N with these options need to be evaluated in light of milk prices and commodity and fertilizer nitrogen costs. At current prices and costs, the economic benefit from the energy supplementation options is substantial.

With school buildings shuttering due to COVID-19, school counselors' work needed to shift to virtual environments. The shift in operations led us to wonder about school counselors' perceptions and practices in these new and unfamiliar conditions. In our mixed-methods cross-sectional study, we gathered quantitative and qualitative data from 89 school counselors. Our findings include evidence of the counselors feeling disconnected from their students, lack of technology and student access, privacy issues, limited preparation to counsel in virtual environments, and work and life balance. We follow our results with a discussion, related implications, and directions for future research.

College students' interactions with campus leaders is critical to their success, particularly in situations of distress. Yet, little is known about college students' knowledge, perceptions, and identification of campus administrators, faculty members, and staff as leaders and their interactions with these campus leaders. To fill the gap in the literature, we applied a cross-sectional methodology to gather a combination of quantitative and qualitative data using an online survey. We had 60 first-year students participate in our exploratory research by fully completing our survey. We found that students identified their advisors as leaders on campus. We also found most of our participants avoided campus administrators in fear of judgment, intimidation, and feelings of anxiety. Our results have implications for campus leadership, college administrators, student retention, and campus climate. Following our results, we discuss implications for practice and offer additional recommendations for future research.

Wildland–urban interfaces (WUIs), the juxtaposition of highly and minimally developed lands, are an increasingly prominent feature on Earth. WUIs are hotspots of environmental and ecological change that are often priority areas for planning and management. A better understanding of WUI dynamics and their role in the coupling between cities and surrounding wildlands is needed to reduce the risk of environmental hazards, ensure the continued provisioning of ecosystem services, and conserve threatened biodiversity. To fill this need, we propose an expanded framework for WUIs that not only conceptualizes these interfaces as emergent and functional components of socioecological processes but also extends them vertically from the bedrock to the top of the vegetation and horizontally across heterogeneous landscapes. This framework encourages management that reconciles pervasive trade-offs between development and resulting multiple environmental impacts. Focusing on southern California as a case study, we use the framework to facilitate integration across disciplines and between scientists and managers.

Our previous results with flight (FLT) mice showed abnormalities in thymuses and spleens that have potential to compromise immune defense mechanisms. In this study, the organs were further evaluated in C57BL/6 mice after Space Shuttle Atlantis returned from a 13-day mission. Thymuses and spleens were harvested from FLT mice and ground controls housed in similar animal enclosure modules (AEM). Organ and body mass, DNA fragmentation and expression of genes related to T cells and cancer were determined. Although significance was not obtained for thymus mass, DNA fragmentation was greater in the FLT group (P

[This corrects the article on p. e75097 in vol. 8.].