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Background Inflammation is a key driver of the transition of acute kidney injury to progressive fibrosis and chronic kidney disease (AKI-to-CKD transition). Blocking a-disintegrin-and-metalloprotease-17 (ADAM17)-dependent ectodomain shedding, in particular of epidermal growth factor receptor (EGFR) ligands and of the type 1 inflammatory cytokine tumor necrosis factor (TNF), reduces pro-inflammatory and pro-fibrotic responses after ischemic AKI or unilateral ureteral obstruction (UUO), a classical fibrosis model. Metalloprotease or EGFR inhibition show significant undesirable side effects in humans. In retrospective studies anti-TNF biologics reduce the incidence and progression of CKD in humans. Whether TNF has a role in AKI-to-CKD transition and how TNF inhibition compares to EGFR inhibition is largely unknown. Methods Mice were subjected to bilateral renal ischemia-reperfusion injury or unilateral ureteral obstruction. Kidneys were analyzed by histology, immunohistochemistry, qPCR, western blot, mass cytometry, scRNA sequencing, and cytokine profiling. Results Here we show that TNF or EGFR inhibition reduce AKI-to-CKD transition and fibrosis equally by about 25%, while combination has no additional effect. EGFR inhibition reduced kidney TNF expression by about 50% largely by reducing accumulation of TNF expressing immune cells in the kidney early after AKI, while TNF inhibition did not affect EGFR activation or immune cell accumulation. Using scRNAseq data we show that TNF is predominantly expressed by immune cells in AKI but not in proximal tubule cells (PTC), and PTC-TNF knockout did not affect AKI-to-CKD transition in UUO. Thus, the anti-inflammatory and anti-fibrotic effects of the anti-TNF biologic etanercept in AKI-to-CKD transition rely on blocking TNF that is released from immune cells recruited or accumulating in response to PTC-EGFR signals. Conclusion Short-term anti-TNF biologics during or after AKI could be helpful in the prevention of AKI-to-CKD transition.

IntroductionPrevious studies demonstrate that a significant proportion of casualties do not receive pain medication prehospital after traumatic injuries. To address possible reasons, the U.S. Military has sought to develop novel delivery methods to aid in administration of pain medications prehospital. We sought to describe the dose and route of ketamine administered prehospital to help inform materiel solutions.Materials and MethodsThis is a secondary analysis of a previously described dataset focused on prehospital data within the Department of Defense Trauma Registry from 2007 to 2020. We isolated encounters in which ketamine was administered along with the amount dosed and the route of administration in nonintubated patients.ResultsWithin our dataset, 862 casualties met inclusion for this analysis. The median age was 28 and nearly all (98%) were male. Most were battle injuries (88%) caused by explosives (54%). The median injury severity score was 10 with the extremities accounting to the most frequent seriously injured body region (38%). The mean dose via intravenous route was 50.4 mg (n = 743, 95% CI 46.5-54.3), intramuscular was 66.7 mg (n = 234, 95% CI 60.3-73.1), intranasal was 56.5 mg (n = 10, 39.1-73.8), and intraosseous was 83.3 mg (n = 34, 66.3-100.4). Most had a medic or CLS in their chain of care (87%) with air evacuation as the primary mechanism of evacuation (86%).ConclusionsThe average doses administered were generally larger than the doses recommended by Tactical Combat Casualty Care guidelines. Currently, guidelines may underdose analgesia. Our data will help inform materiel solutions based on end-user requirements.

BackgroundAdvanced glycation end products (AGEs) are formed via the nonenzymatic glycation of sugars with amino acids. Two AGEs, Nε-(1-carboxymethyl)-L-Lysine (CML) and pentosidine, have been observed to be elevated in subjects suffering from a multitude of chronic disease states, and accumulation of these compounds may be related to the pathophysiology of disease progression and aging.MethodsWe describe here the development and validation of a specific and reproducible LC-MS/MS method to quantify CML and pentosidine in human serum with lower limits of quantitation of 75 ng/mL and 5 ng/mL, respectively. The analyte calibration curve exhibited excellent linearity at a range of 0–10 900 ng/mL for CML and 0–800 ng/mL for pentosidine. High-low linearity of 5 serum pairs was assessed, with a mean recovery of 103% (range 94—116%) for CML, and 104% (range 97—116%) for pentosidine.ResultsSerum concentrations of CML and pentosidine were quantified in 30 control and 30 subjects with chronic renal insufficiency. A significant increase in both analytes was observed in renal failure compared to control subjects (2.1-fold and 8.4-fold, respectively; P ConclusionsThese mass spectroscopy-based assays for serum CML and pentosidine should be useful in accurately evaluating circulating levels of these key AGEs in various disease states.

Societies are organizing themselves to keep the COVID-19 virus at bay for the foreseeable future. The World Health Organization (WHO) has proposed that every country implement a comprehensive set of measures to prevent infection, detect cases, interrupt transmission, control clusters, suppress outbreaks and reduce mortality. Throughout the world, four systems capabilities are emerging that are important for societies to get ahead of the virus and become COVID-19 ready. First: understand the pattern of infection locally and act on it effectively: assess the status of the outbreak; act rapidly and robustly to interrupt transmission. Second: enable people to be active participants in their own responses. Third: focus on the places where people are most at risk of infection. Fourth: assess the performance of responses to COVID-19 infection: detecting cases, interrupting chains of transmission, minimizing adverse consequences, protecting the most vulnerable and ensuring opportunities for sustainable livelihoods and well-being for all, leaving no-one behind. These four capabilities are being woven together within societies: successful weaving can be helped through focusing on three interlinked elements: making information available; assessing and reducing risk; and being able to suppress outbreaks rapidly. This means involving everyone in the response and having strong public health defences. Governments, authorities, public health teams, employers and community organizations make it possible for us to stay safe but, in the end, what happens is up to all of us, individually and collectively. If we are to live well with the threat of COVID-19, solidarity really does matter.

Introduction/Objective Blood cultures are commonly obtained to evaluate the presence of bacteria or fungal infection in a patient’s bloodstream. The presence of living microorganisms circulating in the bloodstream is of substantial prognostic and diagnostic importance. A positive blood culture indicates a reason for the patient’s illness and provides the etiological agent for antimicrobial therapy. Collection of blood culture is an exact process that requires time, the proper order of draw, and following of correct protocol. The busy Emergency department that requires multiple demands for nurse’s time, turnover of staff, rushing from one task to another can result in the improper collection and false-positive blood cultures. The national benchmark is set at 3% by the American Society of Clinical Microbiology (ASM) and The Clinical and Laboratory Standard Institute (CLSI). False-positive blood culture results in increased length of stay and unnecessary antimicrobial therapy, resulting in an increased cost burden to the hospital of about $5000 per patient. Methods/Case Report At our 150-bed community hospital, 26 beds Emergency Department, we have come a long way in reduction of our blood culture contamination rates from upwards of 4% to less than 2%, far lower than the national benchmark. Results (if a Case Study enter NA) NA Conclusion There are multiple devices available from various manufacturers claiming to reduce blood culture contamination. These devices do reduce blood culture (BC) contamination but at an added cost of the device. The rate of BC can be reduced and less than 3% is achievable by materials available in the laboratory. We have achieved this by providing training to every new staff by demonstration and direct observation, providing everything required for collection in a kit, using proper technique, the inclusion of diversion method that involves the aseptic collection of a clear tube before collecting blood cultures, and following up monthly on any false positive blood cultures.

Background Frailty confers risk for surgical morbidity and mortality. Whether patient-reported measures of health, well-being, or quality of life respond differently to surgery in non-frail and frail individuals is unknown. Methods Older adults with severe aortic stenosis presenting for surgery were assessed for frailty using Cardiovascular Health Study Criteria. Patient-reported measures of functional capacity (Duke Activity Status Index [DASI]), physical and mental health (Medical Outcomes Study Short Form-Physical and Mental Component Scales [SF-12 PCS and SF-12 MCS, respectively]), well-being (linear analogue self-assessment [LASA]), and quality of life (LASA) were administered before and 3 months after surgery. Results Of 103 participants (mean age of 80.6 years), 54 were frail. Frail participants had lower baseline DASI, SF-12 PCS, SF-12 MCS, physical well-being, and quality of life scores than non-frail participants. At follow-up, frail participants showed significant improvement in physical function, with DASI and SF-12 PCS scores improving by 50% and 14%, respectively. Non-frail subjects did not significantly improve in these measures. SF-12 MCS scores also improved to a greater extent in frail compared to non-frail participants (3.6 vs < 1 point). Furthermore, the frail participants improved to a greater extent than non-frail participants in physical well-being (21.6 vs 7.1 points) and quality of life measures (25.1 vs 8.7 points). Conclusions Frailty is prevalent in older adults with severe aortic stenosis and is associated with poor physical and mental function, physical well-being, and quality of life. In response to surgery, frail participants exhibited greater improvement in these patient-centered outcomes than non-frail peers.

Being a woman is one of the strongest risk factors for multiple sclerosis. The natural reproductive period from menarche to natural menopause corresponds to the active inflammatory disease period in multiple sclerosis. The fifth decade marks both the peri-menopausal transition in the reproductive aging and a transition from the relapsing-remitting to the progressive phase in multiple sclerosis. A short reproductive period with premature/early menopause and/or low number of pregnancies may be associated with an earlier onset of the progressive multiple sclerosis phase. A cross-sectional study of survey-based reproductive history in a multiple sclerosis clinical series enriched for patients with progressive disease, and a case–control study of multiple sclerosis and age/sex matched controls from a population-based cohort were conducted. Menarche age, number of complete/incomplete pregnancies, menopause type and menopause age were compared between 137 cases and 396 control females. Onset of relapsing-remitting phase of multiple sclerosis, progressive disease onset and reaching severe disability (expanded disability status scale 6) were studied as multiple sclerosis-related outcomes (n = 233). Menarche age was similar between multiple sclerosis and control females (P = 0.306). Females with multiple sclerosis had fewer full-term pregnancies than the controls (P, Premature/early menopause or nulliparity was associated with earlier progressive multiple sclerosis onset with a ‘dose effect’ of pregnancies on delaying progressive multiple sclerosis and severe disability. Zeydan et al.’s findings suggest a beneficial impact of oestrogen in delaying progressive multiple sclerosis and may have implications for counselling women on reproductive health., Graphical Abstract

To determine early somatic changes in high-grade serous ovarian cancer (HGSOC), we performed whole genome sequencing on a rare collection of 16 low stage HGSOCs. The majority showed extensive structural alterations (one had an ultramutated profile), exhibited high levels of p53 immunoreactivity, and harboured a TP53 mutation, deletion or inactivation. BRCA1 and BRCA2 mutations were observed in two tumors, with nine showing evidence of a homologous recombination (HR) defect. Combined Analysis with The Cancer Genome Atlas (TCGA) indicated that low and late stage HGSOCs have similar mutation and copy number profiles. We also found evidence that deleterious TP53 mutations are the earliest events, followed by deletions or loss of heterozygosity (LOH) of chromosomes carrying TP53, BRCA1 or BRCA2. Inactivation of HR appears to be an early event, as 62.5% of tumours showed a LOH pattern suggestive of HR defects. Three tumours with the highest ploidy had little genome-wide LOH, yet one of these had a homozygous somatic frame-shift BRCA2 mutation, suggesting that some carcinomas begin as tetraploid then descend into diploidy accompanied by genome-wide LOH. Lastly, we found evidence that structural variants (SV) cluster in HGSOC, but are absent in one ultramutated tumor, providing insights into the pathogenesis of low stage HGSOC.

Reversible lysine acetylation is a highly regulated post-translational protein modification that is known to regulate several signaling pathways. However, little is known about the radiation-induced changes in the acetylome. In this study, we analyzed the acute post-translational acetylation changes in primary human cardiac microvascular endothelial cells 4 h after a gamma radiation dose of 2 Gy. The acetylated peptides were enriched using anti-acetyl conjugated agarose beads. A total of 54 proteins were found to be altered in their acetylation status, 23 of which were deacetylated and 31 acetylated. Pathway analyses showed three protein categories particularly affected by radiation-induced changes in the acetylation status: the proteins involved in the translation process, the proteins of stress response, and mitochondrial proteins. The activation of the canonical and non-canonical Wnt signaling pathways affecting actin cytoskeleton signaling and cell cycle progression was predicted. The protein expression levels of two nicotinamide adenine dinucleotide (NAD+)-dependent deacetylases, sirtuin 1 and sirtuin 3, were significantly but transiently upregulated 4 but not 24 h after irradiation. The status of the p53 protein, a target of sirtuin 1, was found to be rapidly stabilized by acetylation after radiation exposure. These findings indicate that post-translational modification of proteins by acetylation and deacetylation is essentially affecting the radiation response of the endothelium.

International audience; In 2009, the European High Level and Expert Group identified key policy and scientific questions to be addressed through a strategic research agenda for low-dose radiation risk. This initiated the establishment of a European Research Platform, called MELODI (Multidisciplinary European Low Dose Research Initiative). In 2010, the DoReMi Network of Excellence was launched in the Euratom 7th Framework Programme. DoReMi has acted as an operational tool for the sustained development of the MELODI platform during its early years. A long-term Strategic Research Agenda for European low-dose radiation risk research has been developed by MELODI. Strategic planning of DoReMi research activities is carried out in close collaboration with MELODI. The research priorities for DoReMi are designed to focus on objectives that are achievable within the 6-y lifetime of the project and that are in areas where stimulus and support can help progress towards the longer-term strategic objectives. © The Author 2014.

Senescent cells drive age-related tissue dysfunction through their potent secretome, termed the senescence associated secretory phenotype (SASP). Circulating concentrations of SASP factors may reflect biological age and serve as clinically useful biomarkers of surgical risk and ultimately, surrogate endpoints in clinical trials. However, they remain largely uncharacterized. We tested associations between circulating concentrations of SASP proteins and biological age, as determined by the accumulation of age-related health deficits, and/or postoperative outcomes in a sample of residents in Olmstead County, MN, age 60-90 years (n = 115) and cohorts of older adults undergoing surgery for severe aortic stenosis (prospective; n = 97) or ovarian cancer (case-control; n = 36). Circulating concentrations of SASP factors were associated with biological age and adverse postoperative outcomes, including risk of any adverse event or rehospitalization within the year following surgery (aortic stenosis group) or admission to an intensive care unit within 30 days of hospital discharge (ovarian cancer group). Gradient boosting machine modeling revealed a panel of SASP factors that predicted adverse outcomes across both surgical groups better than biological age or chronological age and sex. This suggests that the circulating SASP is a robust indicator of age-related health status and may help guide clinical decision making. Furthermore, circulating SASP factors may be harnessed as a readily quantifiable biomarkers in senescence-targeting interventional human studies.

A nation's opioid consumption was once seen as a sign of civility. The higher the per capita rate of morphine consumption, as calculated by the World Health Organization, the more advanced and caring a nation's health care system was seen to be—this suggested better care of the elderly, the sick, the suffering, and the dying [1]. So it is of interest that when numbers from IMS Health were released earlier this year (and reported in a feature article in the Milwaukee Journal Sentinel) showed that opioid prescribing has experienced a slight decrease from 243 million prescriptions in 2011 to 241 million in 2012, this change was hailed as a sign of good things to come for Americans [2]. While a reduction in “pill mill” activity and inappropriate prescribing should be lauded, there is unfortunately much to be concerned about for both the majority of people with pain and the well-intentioned prescribers alike. Moreover, the sensationalization by mainstream media and by politicians has served to fuel a fire that creates negative outcomes for legitimate pain patients [3,4]. There is no question that at one time it was suggested that more opioids were better, while to some extent minimizing the risks to individual patients and to society at large. There is still some debate over the reasons for the rapid growth of opioid use. We argue that this growth was largely due to well-intentioned physicians and thought leaders who had positive experiences treating pain in the advanced cancer patient population. Expanded opioid prescribing in that arena led to dramatic improvements in quality of life, and as a result, many were hopeful that similar improvements would be seen in the much larger and more diverse chronic noncancer pain population [5–7]. When reports of OxyContin abuse first surfaced over a …

In field conditions, plants may experience numerous environmental stresses at any one time. Research suggests that the plant response to multiple stresses is different from that for individual stresses, producing nonadditive effects. In particular, the molecular signaling pathways controlling biotic and abiotic stress responses may interact and antagonize one another. The transcriptome response of Arabidopsis (Arabidopsis thaliana) to concurrent water deficit (abiotic stress) and infection with the plant-parasitic nematode Heterodera schachtii (biotic stress) was analyzed by microarray. A unique program of gene expression was activated in response to a combination of water deficit and nematode stress, with 50 specifically multiple-stress-regulated genes. Candidate genes with potential roles in controlling the response to multiple stresses were selected and functionally characterized. RAPID ALKALINIZATION FACTOR-LIKE8 (AtRALFL8) was induced in roots by joint stresses but conferred susceptibility to drought stress and nematode infection when overexpressed. Constitutively expressing plants had stunted root systems and extended root hairs. Plants may produce signal peptides such as AtRALFL8 to induce cell wall remodeling in response to multiple stresses. The methionine homeostasis gene METHIONINE GAMMA LYASE (AtMGL) was up-regulated by dual stress in leaves, conferring resistance to nematodes when overexpressed. It may regulate methionine metabolism under conditions of multiple stresses. AZELAIC ACID INDUCED1 (AZI1), involved in defense priming in systemic plant immunity, was down-regulated in leaves by joint stress and conferred drought susceptibility when overexpressed, potentially as part of abscisic acid-induced repression of pathogen response genes. The results highlight the complex nature of multiple stress responses and confirm the importance of studying plant stress factors in combination.

Background and Aims: Understanding the synthesis of ascorbic acid (l-AsA) in green tissues in model species has advanced considerably; here we focus on its production and accumulation in fruit. In particular, our aim is to understand the links between organs which may be sources of l-AsA (leaves) and those which accumulate it (fruits). The work presented here tests the idea that changes in leaf and fruit number influence the accumulation of l-AsA. The aim was to understand the importance of leaf tissue in the production of l-AsA and to determine how this might provide routes for the manipulation of fruit tissue l-AsA. Methods: The experiments used Ribes nigrum (blackcurrant), predominantly in field experiments, where the source–sink relationship was manipulated to alter potential leaf l-AsA production and fruit growth and accumulation of l-AsA. These manipulations included reductions in reproductive capacity, by raceme removal, and the availability of assimilates by leaf removal and branch phloem girdling. Natural variation in fruit growth and fruit abscission is also described as this influences subsequent experimental design and the interpretation of l-AsA data. Key Results: Results show that fruit l-AsA concentration is conserved but total yield of l-AsA per plant is dependent on a number of innate factors many of which relate to raceme attributes. Leaf removal and phloem girdling reduced fruit weight, and a combination of both reduced fruit yields further. It appears that around 50 % of assimilates utilized for fruit growth came from apical leaves, while between 20 and 30 % came from raceme leaves, with the remainder from ‘storage’. Conclusions: Despite being able to manipulate leaf area and therefore assimilate availability and stored carbohydrates, along with fruit yields, rarely were effects on fruit l-AsA concentration seen, indicating fruit l-AsA production in Ribes was not directly coupled to assimilate supply. There was no supporting evidence that l-AsA production occurred predominantly in green leaf tissue followed by its transfer to developing fruits. It is concluded that l-AsA production occurs predominantly in the fruit of Ribes nigrum.

Co-ordination of metabolic and physiological activity between plant parts is key to the control of growth and development. Here the movement of resources and their allocation between mother plants and daughter ramets along Fragaria stolons was quantified with respect to hierarchy. Gradients of internodal ramet leaf water potential (ψ) and stolon and ramet hydraulic conductivities (L) were measured together with apparent stolon IAA movement via the polar auxin transport pathway (PAT). These processes are linked with measurements of stolon vascular development. The pattern of tissue differentiation and lignification in sequential stele sections of stolons demonstrated the rapid acquisition of the capacity for water transport, with transpiration potentially varying systematically with stolon lignification and the acropetal decline in stolon xylem ψ. Stolon and ramet L declined acropetally, with L across older ramets being significantly lower than that of the connecting stolons. The capacity for polar IAA transport increased with stolon age; this was due to increased transport intensity in older tissues. The partitioning of dry matter was strongly hierarchical with younger ramets smaller than older ramets, while foliar concentrations of N, P, and K were greater for the younger ramets. The results show that stolon anatomy develops rapidly at the apical end, facilitating hierarchical ramet development, which is evident as a basipetal increase in L. The rapid development of transport tissue functionality enables young unrooted ramets to acquire water, in order to supply an expanding leaf area, as well as mineral ions disproportionally with respect to older ramets. This facilitates colonization and self-rooting of apical ramets. The unidirectional increase in basipetal PAT along stolons facilitates hierarchical ramet development.

Plant responses to different stresses are highly complex and involve changes at the transcriptome, cellular, and physiological levels. Recent evidence shows that plants respond to multiple stresses differently from how they do to individual stresses, activating a specific programme of gene expression relating to the exact environmental conditions encountered. Rather than being additive, the presence of an abiotic stress can have the effect of reducing or enhancing susceptibility to a biotic pest or pathogen, and vice versa. This interaction between biotic and abiotic stresses is orchestrated by hormone signalling pathways that may induce or antagonize one another, in particular that of abscisic acid. Specificity in multiple stress responses is further controlled by a range of molecular mechanisms that act together in a complex regulatory network. Transcription factors, kinase cascades, and reactive oxygen species are key components of this cross-talk, as are heat shock factors and small RNAs. This review aims to characterize the interaction between biotic and abiotic stress responses at a molecular level, focusing on regulatory mechanisms important to both pathways. Identifying master regulators that connect both biotic and abiotic stress response pathways is fundamental in providing opportunities for developing broad-spectrum stress-tolerant crop plants.

Genetic factors can modify susceptibility to the carcinogenic effect of ionising radiation. To establish if radioiodine-induced thyroid cancer is similarly genetically influenced, we studied F1 hybrid crosses between inbred mouse strains. Mice were perinatally exposed to iodine-131 and thyroid tissues examined after 18 months. Differences in the incidence and distribution of histological subtypes were quantified in relation to genetic background. As expected, the occurrence of thyroid lesions was significantly higher in irradiated mouse hybrids than in unirradiated controls. The most frequent alterations were the simple and the complex hyperplasias, followed by follicular adenoma and, less frequently, follicular carcinoma. Both the incidence and distribution of the histiotype were different between the hybrid mouse crosses. Crosses using JF1 mice (M. m. molossinus) produced F1 offspring that were more resistant to radiation-induced thyroid lesions. Sequence analysis of Braf, Ret, Hras, Kras, Kit and Trp53, all genes that are commonly mutated in human thyroid cancers, did not show any evidence of mutation in the tumours. However, microsatellite analysis of genomic DNA revealed frequent allelic imbalances in complex hyperplasia and follicular adenoma. We conclude that genetic background, in particular the JF1 genotype, confer differences in susceptibility to the carcinogenic effects of radioiodine on the thyroid.

Background and Aims: The resurgence of malaria, particularly in the developing world, is considerable and exacerbated by the development of single-gene multi-drug resistances to chemicals such as chloroquinone. Drug therapies, as recommended by the World Health Organization, now include the use of antimalarial compounds derived from Artemisia annua – in particular, the use of artemisinin-based ingredients. Despite our limited knowledge of its mode of action or biosynthesis there is a need to secure a supply and enhance yields of artemisinin. The present study aims to determine how plant biomass can be enhanced while maximizing artemisinin concentration by understanding the plant's nutritional requirements for nitrogen and potassium. Methods: Experiments were carried out, the first with differing concentrations of nitrogen, at 6, 31, 56, 106, 206 or 306 mg L–1 being applied, while the other differing in potassium concentration (51, 153 or 301 mg L–1). Nutrients were supplied in irrigation water to plants in pots and after a growth period biomass production and leaf artemisinin concentration were measured. These data were used to determine optimal nutrient requirements for artemisinin yield. Key Results: Nitrogen nutrition enhanced plant nitrogen concentration and biomass production successively up to 106 mg N L–1 for biomass and 206 mg N L–1 for leaf nitrogen; further increases in nitrogen had no influence. Artemisinin concentration in dried leaf material, measured by HPLC mass spectroscopy, was maximal at a nitrogen application of 106 mg L–1, but declined at higher concentrations. Increasing potassium application from 51 to 153 mg L–1 increased total plant biomass, but not at higher applications. Potassium application enhanced leaf potassium concentration, but there was no effect on leaf artemisinin concentration or leaf artemisinin yield. Conclusions: Artemisinin concentration declined beyond an optimal point with increasing plant nitrogen concentration. Maximization of artemisinin yield (amount per plant) requires optimization of plant biomass via control of nitrogen nutrition.

Objective. Equianalgesic conversion methods are commonly used to switch patients from one opioid to another due to suboptimal pain relief or adverse events. There is no universally accepted opioid conversion method, however, and there is often significant variability between conversion resources. As a result, patients are at risk for undertreated pain and serious adverse events. The purpose of this survey was to compare the equianalgesic conversion estimates between nurse practitioners, pharmacists, and physicians for commonly prescribed opioids. Methods. A survey form was developed using Survey Monkey. Participation was solicited by providing a link to the survey via social media (e.g., Facebook, Twitter, LinkedIn, etc.) and emailing professional organizations for sharing with their members and followers. Data collected included demographics and estimated morphine equivalents (MEQs) of hydrocodone 80 mg, fentanyl transdermal patches 1,800 mcg (as 75 mcg/hour), methadone 40 mg, oxycodone 120 mg, and hydromorphone 48 mg. Participants were also asked to provide their choice of reference utilized to complete the conversions, including personal knowledge. Descriptive analyses were performed using measures of central tendency. Hypothesis testing was performed using Pearson's chi-squared and Fisher's Exact Test for categorical data and the Kruskal–Wallis equality of populations rank test for continuous data to assess differences between median opioid doses by professional groups. Results. The total number of respondents included in the analysis was 319. Physicians, pharmacists, and nurse practitioners/physician assistants comprised 25.4%, 56.7%, and 16.3%, respectively, of respondents. The overall mean (± standard deviation) MEQ doses for fentanyl, hydrocodone, hydromorphone, methadone, and oxycodone were: 176 (±117) mg, 88 (±42) mg, 192 (±55) mg, 193 (±201) mg, and 173 (±39) mg, respectively. For fentanyl, the mean (±standard deviation) MEQ doses were 180 (±122) mg, 178 (±128) mg, and 157 (±68) mg, for physicians, pharmacists, and nurse practitioners/physician assistants, respectively. For all three groups of clinicians, the median MEQ dose for fentanyl was 150 mg. The mean (±standard deviation) MEQ doses of methadone for physicians, pharmacists, and nurse practitioners/physician assistants were: 214 (±142) mg, 171 (±107) mg, and 185 (±129) mg, respectively. The median MEQ dose for methadone was 160 mg for each of the clinician groups. Conclusions. As evidenced by large standard deviations, there was significant variation in mean opioid conversions to MEQ doses within each profession type, particularly for fentanyl and methadone. The median MEQ doses provided for opioid conversions were the same among each profession. No universal method exists that allows each of the five studied opioids to be accurately and consistently converted to another opioid (i.e., morphine).

• Background and Aims A number of strawberry varieties were surveyed for their total ellagic acid concentration, and attempts were made to determine if ellagic acid and ascorbic acid concentrations of two strawberry cultivars could be increased by polythene reflective mulches. • Methods After adjusting crop yields and cultivation using polythene mulches with two different PAR reflective capacities, field- and polytunnel-grown strawberries were analysed for ellagic acid and ascorbic acid concentrations by HPLC. Comparative measurements of yield and fruit quality were determined along with plant developmental changes. • Key Results Ellagic acid concentration varied widely with strawberry cultivar (60–341µg g−1 frozen weight), as did the ratio of conjugated ellagic acid:free ellagic acid. Also, there was significant year-to-year variation in total ellagic acid concentration with some cultivars. Mulches with different reflective capacities impacted on strawberry production; highly reflective mulches significantly increased growth and yield, the latter due to increases in fruit size and number. • Conclusions Highly reflective mulches significantly increased total concentrations of ellagic acid and ascorbic acid relative to control in fruit of different cultivars. The potential of agronomic practices to enhance the concentration and amounts of these important dietary bioactive compounds is discussed.

It was investigated whether premature fruit abscission in Prunus avium L. was triggered by a reduction in polar auxin transport (PAT). The capacity of pedicels to transport tritiated IAA ([ 3 H]-IAA) via the PAT pathway was measured at intervals throughout flower and fruit development. The extent of passive diffusion, assessed by concurrent applications of [ 14 C]-benzoic acid ([ 14 C]-BA), was negligible. Transported radioactivity recovered from agar blocks eluted at the same retention time as authentic [ 3 H]-IAA during HPLC fractionation. The capacity for PAT was already high 7 d before anthesis and increased further following the fertilization of flowers at anthesis. PAT intensity was greatest immediately following fertilization and at the beginning of the cell expansion phase of fruit growth; the transport intensity in fruitlets destined to abscind was negligible. The amount of endogenous IAA moving through the PAT pathway was greatest during the first 3 weeks after fertilization and was again high at the beginning of the fruit expansion stage. IAA export in the phloem increased following fertilization then declined below detectable levels. ABA export in the phloem increased markedly during stone formation and at the onset of fruit expansion. TIBA applied to pedicels of fruit in situ promoted fruitlet abscission in 2000 but not in 2001, despite PAT capacity being reduced by over 98% in the treated pedicels. The application of TIBA to pedicels did not affect fruit expansion. The role of PAT and IAA in relation to the development and retention of Prunus avium fruit is discussed.

We have recently identified a locus exhibiting a high frequency of allelic imbalance (AI) in both spontaneous human (HSA 6q14.1-15) and radiogenic murine (MMU9, 42 cM) osteosarcoma. Here we describe the fine mapping of the locus in osteosarcoma arising in (BALB/c x CBA) F 1 hybrid mice. These studies have allowed us to identify Tbx18, a member of the T-box transcriptional regulator gene family, as a candidate gene. Three intragenic Tbx18 polymorphisms were used to map the region of maximum AI to within the gene itself; 16 of 17 tumours exhibited imbalances of at least one of these markers. The highest frequency was found in exon 1, where 14 of 17 tumours were affected at a single nucleotide polymorphism at 541 nt. Two polymorphic CA repeat markers in intron 2 and intron 5 demonstrated overlapping regions of imbalance in several tumours. Both markers flanking the Tbx18 gene (D9Osm48 and D9Mit269) revealed significantly lower frequencies of imbalance and confirmed the limitation of the common interval to Tbx18. Examination of both the mouse and human annotated genomic sequences indicated Tbx18 to be the only gene within the interval. Sequence analysis of the Tbx18 coding region did not reveal any evidence of mutation. Given the haploinsufficiency phenotypes reported for other T-box genes, we speculate that AI may influence the function of Tbx18 during osteosarcomagenesis.

BACKGROUND: A randomized trial conducted by the Gynecologic Oncology Group (GOG, study #111) in the United States showed a better outcome for patients with advanced ovarian cancer on the paclitaxel-cisplatin regimen than for those on a standard cyclophosphamide-cisplatin regimen. Before considering the paclitaxel-cisplatin regimen as the new "standard," a group of European and Canadian investigators planned a confirmatory phase III trial. METHODS: This intergroup trial recruited 680 patients with broader selection criteria than the GOG #111 study and administered paclitaxel as a 3-hour instead of a 24-hour infusion; progression-free survival was the primary end point. Patient survival was analyzed by use of the Kaplan-Meier technique. Treatment effects on patient survival were estimated by Cox proportional hazards regression models. All statistical tests were two-sided. RESULTS: The overall clinical response rate was 59% in the paclitaxel group and 45% in the cyclophosphamide group; the complete clinical remission rates were 41% and 27%, respectively; both differences were statistically significant (P =.01 for both). At a median follow-up of 38.5 months and despite a high rate of crossover (48%) from the cyclophosphamide arm to the paclitaxel arm at first detection of progression of disease, a longer progression-free survival (log-rank P =.0005; median of 15.5 months versus 11.5 months) and a longer overall survival (log-rank P =. 0016; median of 35.6 months versus 25.8 months) were seen in the paclitaxel regimen compared with the cyclophosphamide regimen. CONCLUSIONS: There is strong and confirmatory evidence from two large randomized phase III trials to support paclitaxel-cisplatin as the new standard regimen for treatment of patients with advanced ovarian cancer.

Production cross sections of nitrogen isotopes from high-energy carbon isotopes on hydrogen and carbon targets have been measured for the first time for a wide range of isotopes. The fragment separator FRS at GSI was used to deliver C isotope beams. The cross sections of the production of N isotopes were determined by charge measurements of forward going fragments. The cross sections show a rapid increase with the number of neutrons in the projectile. Since the production of nitrogen is mostly due to charge exchange reactions below the proton separation energies, the present data suggests a concentration of Gamow-Teller and Fermi transition strength at low excitation energies for neutron-rich isotopes. It was also observed that the cross sections were enhanced much more strongly for neutron rich isotopes in the C-target data., Comment: 11 pages, 9 figures, 30 references

Cystatins are small protein inhibitors of cysteine proteinases. The relative importance of the N-terminal region of cystatins, and of a conserved glycine within this region, remains unclear despite several studies. It was found that deletion of the N-terminal 21 amino acids abolishes the inhibitory capacity of oryzacystatin-I. The importance of a conserved glycine residue (Gly10) was also examined by replacing it with 11 other amino acids. Three further glycine residues (Gly5, -6 and -11) in this N-terminal region of oryzacystatin-I were similarly mutated. Only those variants in which Gly10 was substituted show any significant change in inhibitory capacity compared with wild-type oryzacystatin-I. The inhibitory characteristics of hybrid cystatin molecules comprising regions of chicken egg white cystatin and oryzacystatin were also examined. It is suggested that in common with animal cystatins, the N-terminal region of the plant cystatin, oryzacystatin-I, and in particular the highly conserved Gly10 residue are important for effective inhibition of papain.

A detailed study of the calcifuge Lupinus Iuteus L. (yellow lupin) has been carried out in an attempt to explain its poor performance in the presence of high concentrations of rhizosphenc calcium. Plants were grown on two different calcium regimes, 1 or 15 mol m– Ca and, after an establishment period, measurements were made of the rate of leaf extension, final length of the leaflets and the leaf gas exchange. In addition, the distribution of calcium within the leaf tissue was investigated. At 15 mol m–3 Ca, leaflet length at full expansion was reduced as a consequence of reduced extension rate and a decline in cell wall extensibility. Transpiration in excised leaves, assayed gravimetrically, was significantly reduced in plants grown in high calcium. Similar results were also obtained from gas exchange measurements. Analysis of A/C, curves indicated that in plants grown in high [Ca] there was a substantial reduction in net assimilation over a range of concentrations of CO2 X-ray microanalysis revealed that a large amount of cal cium delivered in the xylem sap is retained in the mesophyll tissue, and most of that reaching the epidermal tissue is not found in the guard cells but in the cells adjacent to them, which in this species are not anatomically distinct as ‘subsidiary’ cells.

Individual genetic variation can influence susceptibility to the carcinogenic effects of many environmental carcinogens. In radiation-exposed populations those individuals with a greater genetically determined susceptibility would be at greater risk of developing cancer. To include this modification of risk into radiation protection schemes it is necessary to identify the genes responsible for determining individual sensitivity. Alpha-particle-induced osteosarcoma in the mouse has been adopted as a model of human radiation carcinogenesis, and genome-wide screens have been conducted for allelic imbalance and genetic linkage. These studies have revealed a series of genes involved in determining the sensitivity to radiogenic osteosarcoma formation.

Vertebral fractures are the most frequent of the fractures associated with osteoporosis, yet little is known of their impact on health in the United States. To aid in this understanding, the authors examined the survival rate of 335 residents of Rochester, Minnesota, who had an initial radiologic diagnosis of vertebral fracture between 1985 and 1989. Seventy-six died during 809 person-years of follow-up. The overall survival rate was worse than expected, and diverged steadily from expected values throughout the course of the study. At 5 years after diagnosis, the estimated survival was 61% compared with an expected value of 76% (relative survival = 0.81, 95% confidence interval (CI) 0.70-0.92). The 5-year relative survival after a hip fracture in Rochester was a comparable 0.82 (95% CI 0.77-0.87), but there was a much greater excess of deaths within the first 6 months as compared with patients with vertebral fractures. The 5-year relative survival rate after a distal forearm fracture was 1.00 (95% CI 0.95-1.05). Clinically diagnosed vertebral fractures are rarely fatal, and the reduced survival seen subsequently could related to comorbid conditions. Nonetheless, the excess mortality should be accounted for in assessing the public health impact of osteoporosis.

Morphological and physiological characteristics of micropropagated plants of Delphilium cv. Princess Caroline were studied. Leaves produced in vitro showed poor control of water loss which appears to result from restricted responses by stomata and not from poor cuticular development. Stomata of leaves produced in vitro were larger and more frequent than those produced during acclimatization. Despite the fact that stomata from isolated epidermis of leaves produced in vitro reduced their apertures when exposed to turgor-reducing treatments, they did not close fully

The primary repertoire in mice, in large part, is determined by the Ig gene segments joined to form the variable region genes-VDJ for the heavy chain genes and VJ for the light chain genes. However, the mechanisms that determine which VH gene, of several hundred available, is joined to a DJH structure remain unexplained. One theory proposes that the VH gene segments closest to the 3' end of the VH locus are chosen because of their location, i.e. that proximity to a DJH structure is a prime determinant for VH selection. Alternatively, the content of the DNA may be the determining factor regardless of chromosomal position. According to this hypothesis 3' preference is due to the coincidence of preferred content and proximity to DJH structures. Since the 3' VH families span more than a centimorgan of DNA we were able to distinguish between these concepts by determining VH usage of gene segments within this region. The BALB/c strain is ideal for this study since it uses 3' VH families at a very high frequency. We therefore mapped the positions of the 3' VH gene families and used this map to investigate the usage of these gene segments in BALB/c fetal liver derived A-MuLV transformed cell lines. We found that while VH gene segments throughout the Ig locus, both 3' and 5' were used, there was a clear preference for 3' VH families. However, within the 3' VH segments the most proximal were not preferred, indicating that proximity to DJH structure was not a strict determinant of VH utilization.

Amounts of total and free calcium in root and shoot xylem sap were quantified for a number of species grown in comparable environments and in a rooting medium not deficient in calcium. The potential for the shoot to sequester calcium was also examined, along with the ability for ABA to regulate calcium flux to the leaf. Xylem sap calcium showed considerable interspecific and diurnal variation, even though the plants were grown with similar rhizospheric calcium concentrations. The potential for the shoot to sequester xylem sap calcium was also highly variable between species and implied an ability, at least in some species, to regulate the calcium reaching the shoot in the transpiration stream. Long distance transport of calcium in the xylem was not primarily by mass flow, because neither calcium uptake nor distribution were closely related to water uptake or transpiration. The diurnal changes in xylem sap total ion concentration appeared to be negatively correlated with transpiration while, in contrast, the calcium ion concentration showed two peaks, one occurring in the dark and the other in the light period. The application of ABA to roots caused an increase in the rate of exudation from the xylem of detopped well-watered plants. These experiments suggest that changes in root water relations driven by ionic fluxes were the likely cause for enhanced sap exudation from ABA-treated roots. The steady-state concentration of calcium in the xylem sap was unaffected by ABA when exudation rate increased and, consequently, the flux of calcium must also have increased.

The mechanism of S02-induced changes in stomatal conductance (g) of alder was examined to determine if S02 affects guard cell function directly or indirectly through the S02-induced changes in photosynthesis. During experimental fumigations at S02 concentrations of 3-3 /?mol m"3 (0 08 /J 1 " stomatal closure preceded declines in net photosynthetic rate (A), indicating that S02 can directly affect guard cells. From these and other studies it appears that the sequence of A and g responses may be influenced by S02 concentration as well as by species. Fumigation with S02 did not cause increases in g, even when the intercellular substomatal C02 concentration (ci) was reduced by 50 ??mol mol"1. Increases in g are not attributable to S02 effects on the C02-based stomatal control system.

A combination of high-performance-liquidchromatography (HPLC), sensitive radioimmunoassays and a homologous in vitro bioassay was used to characterise human parathyroid hormone (hPTH)-peptides in human parathyroid adenoma and plasma. Chromatography of several synthetic hPTH-peptides allows the calibration of the HPLC column. On the basis of sequence hydrophobicity the elution position of peptides can be predicted. A model for the determination of the minimal peptide sequence of each peptide has been developed which based on immunological and physicochemical properties allows the characterisation of unknown hPTH-peptides. Using this technique the heterogeneity of circulating hPTH-peptides in human plasma has been examined. Plasma extracts from healthy individuals, osteoporotic, hyperparathyroid and pseudohyperparathyroid patients were investigated. A uniform pattern in the heterogeneity of hPTH-peptides was detected. Using parathyroid adenoma as reference disease specific changes were characterised.

The characteristics of ABA-induced changes in the fluxes of C02 and water vapour from whole leaves of spring wheat (Triticum aestivum cv. Wembley) were examined. Aqueous solutions of ABA were supplied via the transpiration stream to intact leaves of different ages mounted within a gas exchange cuvette. ABA caused a reduction in stomatal conductance (g) that was proportional to the concentration in the solution fed to the leaf. For the maintenance of a reduction in g there was a requirement for a continual supply of ABA. At concentrations greater than 1CT2 mol m-3 ABA reduced g by at least 50% of the control value, while 1-0 mol m-3 closed stomata within 2 h. Concentrations as low as 10~3 mol m-3 produced a 20% reduction in g. As leaves aged they became less responsive to applied ABA. The possibility that the stomatal response may change after a leaf has previously experienced a pulse of ABA was explored by repeating the exposure of a leaf to 10""2 mol m~3 ABA. The first pulse of ABA produced a greater reduction in g than a subsequent exposure the following day. This decline in response of g to ABA on repeated exposure was maintained with leaves of different ages. The characteristics of the stomatal response to ABA are discussed in the context of what is known about the location of receptors for the hormone. It seems likely that a failure to respond to ABA that has previously accumulated in the guard cells should be viewed by means of maximizing the sensitivity to the current supply of ABA. It is suggested that the smaller response of the stomata of older leaves to ABA makes them more susceptible to water stress, so that they can act as sensors for declining water potentials to give early protection to younger, metabolically active leaves.

In approaching an economic problem from the ecological viewpoint, as is so often necessary in investigations of insect pests, a study of the part played by natural control factors is generally of great practical importance as well as of scientific interest. A knowledge of the factors involved and their mode of operation explains the fluctuations in numbers of the pest and should eventually provide a means of anticipating developments. These considerations give a dual bearing to the study of the more important biotic factors affecting the principal cutworm of Saskatchewan, the red-backed cutworm (Euxoa ochrogaster Gn.). The data presented in this preliminary report have been accumulated during the four years 1923 to 1926 and, while the details relate in the main to conditions at Saskatoon, the principles brought out apply throughout the province of Saskatchewan.