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The Alcohol Use Disorders Identification Test (AUDIT) has been developed from a six-country WHO collaborative project as a screening instrument for hazardous and harmful alcohol consumption. It is a 10-item questionnaire which covers the domains of alcohol consumption, drinking behaviour, and alcohol-related problems. Questions were selected from a 150-item assessment schedule (which was administered to 1888 persons attending representative primary health care facilities) on the basis of their representativeness for these conceptual domains and their perceived usefulness for intervention. Responses to each question are scored from 0 to 4, giving a maximum possible score of 40. Among those diagnosed as having hazardous or harmful alcohol use, 92% had an AUDIT score of 8 or more, and 94% of those with non-hazardous consumption had a score of less than 8. AUDIT provides a simple method of early detection of hazardous and harmful alcohol use in primary health care settings and is the first instrument of its type to be derived on the basis of a cross-national study.

These statements about the importance of effect sizes were made by two of the most influential statistician-researchers of the past half-century. Yet many submissions to Journal of Graduate Medical Education omit mention of the effect size in quantitative studies while prominently displaying the P value. In this paper, we target readers with little or no statistical background in order to encourage you to improve your comprehension of the relevance of effect size for planning, analyzing, reporting, and understanding education research studies.In medical education research studies that compare different educational interventions, effect size is the magnitude of the difference between groups. The absolute effect size is the difference between the average, or mean, outcomes in two different intervention groups. For example, if an educational intervention resulted in the improvement of subjects' examination scores by an average total of 15 of 50 questions as compared to that of another intervention, the absolute effect size is 15 questions or 3 grade levels (30%) better on the examination. Absolute effect size does not take into account the variability in scores, in that not every subject achieved the average outcome.In another example, residents' self-assessed confidence in performing a procedure improved an average of 0.4 point on a Likert-type scale ranging from 1 to 5, after simulation training. While the absolute effect size in the first example appears clear, the effect size in the second example is less apparent. Is a 0.4 change a lot or trivial? Accounting for variability in the measured improvement may aid in interpreting the magnitude of the change in the second example.Thus, effect size can refer to the raw difference between group means, or absolute effect size, as well as standardized measures of effect, which are calculated to transform the effect to an easily understood scale. Absolute effect size is useful when the variables under study have intrinsic meaning (eg, number of hours of sleep). Calculated indices of effect size are useful when the measurements have no intrinsic meaning, such as numbers on a Likert scale; when studies have used different scales so no direct comparison is possible; or when effect size is examined in the context of variability in the population under study.Calculated effect sizes can also quantitatively compare results from different studies and thus are commonly used in meta-analyses.The effect size is the main finding of a quantitative study. While a P value can inform the reader whether an effect exists, the P value will not reveal the size of the effect. In reporting and interpreting studies, both the substantive significance (effect size) and statistical significance (P value) are essential results to be reported.For this reason, effect sizes should be reported in a paper's Abstract and Results sections. In fact, an estimate of the effect size is often needed before starting the research endeavor, in order to calculate the number of subjects likely to be required to avoid a Type II, or β, error, which is the probability of concluding there is no effect when one actually exists. In other words, you must determine what number of subjects in the study will be sufficient to ensure (to a particular degree of certainty) that the study has acceptable power to support the null hypothesis. That is, if no difference is found between the groups, then this is a true finding.Statistical significance is the probability that the observed difference between two groups is due to chance. If the P value is larger than the alpha level chosen (eg, .05), any observed difference is assumed to be explained by sampling variability. With a sufficiently large sample, a statistical test will almost always demonstrate a significant difference, unless there is no effect whatsoever, that is, when the effect size is exactly zero; yet very small differences, even if significant, are often meaningless. Thus, reporting only the significant P value for an analysis is not adequate for readers to fully understand the results.For example, if a sample size is 10 000, a significant P value is likely to be found even when the difference in outcomes between groups is negligible and may not justify an expensive or time-consuming intervention over another. The level of significance by itself does not predict effect size. Unlike significance tests, effect size is independent of sample size. Statistical significance, on the other hand, depends upon both sample size and effect size. For this reason, P values are considered to be confounded because of their dependence on sample size. Sometimes a statistically significant result means only that a huge sample size was used.3A commonly cited example of this problem is the Physicians Health Study of aspirin to prevent myocardial infarction (MI).4 In more than 22 000 subjects over an average of 5 years, aspirin was associated with a reduction in MI (although not in overall cardiovascular mortality) that was highly statistically significant: P < .00001. The study was terminated early due to the conclusive evidence, and aspirin was recommended for general prevention. However, the effect size was very small: a risk difference of 0.77% with r2 = .001—an extremely small effect size. As a result of that study, many people were advised to take aspirin who would not experience benefit yet were also at risk for adverse effects. Further studies found even smaller effects, and the recommendation to use aspirin has since been modified.Depending upon the type of comparisons under study, effect size is estimated with different indices. The indices fall into two main study categories, those looking at effect sizes between groups and those looking at measures of association between variables (table 1). For two independent groups, effect size can be measured by the standardized difference between two means, or mean (group 1) – mean (group 2) / standard deviation.The denominator standardizes the difference by transforming the absolute difference into standard deviation units. Cohen's term d is an example of this type of effect size index. Cohen classified effect sizes as small (d = 0.2), medium (d = 0.5), and large (d ≥ 0.8).5 According to Cohen, “a medium effect of .5 is visible to the naked eye of a careful observer. A small effect of .2 is noticeably smaller than medium but not so small as to be trivial. A large effect of .8 is the same distance above the medium as small is below it.” 6 These designations large, medium, and small do not take into account other variables such as the accuracy of the assessment instrument and the diversity of the study population. However these ballpark categories provide a general guide that should also be informed by context.Between group means, the effect size can also be understood as the average percentile distribution of group 1 vs. that of group 2 or the amount of overlap between the distributions of interventions 1 and 2 for the two groups under comparison. For an effect size of 0, the mean of group 2 is at the 50th percentile of group 1, and the distributions overlap completely (100%)—that is , there is no difference. For an effect size of 0.8, the mean of group 2 is at the 79th percentile of group 1; thus, someone from group 2 with an average score (ie, mean) would have a higher score than 79% of the people from group 1. The distributions overlap by only 53% or a non-overlap of 47% in this situation (table 2).5,6Statistical power is the probability that your study will find a statistically significant difference between interventions when an actual difference does exist. If statistical power is high, the likelihood of deciding there is an effect, when one does exist, is high. Power is 1-β, where β is the probability of wrongly concluding there is no effect when one actually exists. This type of error is termed Type II error. Like statistical significance, statistical power depends upon effect size and sample size. If the effect size of the intervention is large, it is possible to detect such an effect in smaller sample numbers, whereas a smaller effect size would require larger sample sizes. Huge sample sizes may detect differences that are quite small and possibly trivial.Methods to increase the power of your study include using more potent interventions that have bigger effects, increasing the size of the sample/subjects, reducing measurement error (use highly valid outcome measures), and raising the α level but only if making a Type I error is highly unlikely.Before starting your study, calculate the power of your study with an estimated effect size; if power is too low, you may need more subjects in the study. How can you estimate an effect size before carrying out the study and finding the differences in outcomes? For the purpose of calculating a reasonable sample size, effect size can be estimated by pilot study results, similar work published by others, or the minimum difference that would be considered important by educators/experts. There are many online sample size/power calculators available, with explanations of their use (BOX).7,8Power must be calculated prior to starting the study; post-hoc calculations, sometimes reported when prior calculations are omitted, have limited value due to the incorrect assumption that the sample effect size represents the population effect size.Of interest, a β error of 0.2 was chosen by Cohen, who postulated that an α error was more serious than a β error. Therefore, he estimated the β error at 4 times the α: 4 × 0.05 = 0.20. Although arbitrary, as this has been copied by researchers for decades, use of other levels will need to be explained.Effect size helps readers understand the magnitude of differences found, whereas statistical significance examines whether the findings are likely to be due to chance. Both are essential for readers to understand the full impact of your work. Report both in the Abstract and Results sections.

MOTIVATION: Molecular biotechnology now makes it possible to build elaborate systems models, but the systems biology community needs information standards if models are to be shared, evaluated and developed cooperatively. RESULTS: We summarize the Systems Biology Markup Language (SBML) Level 1, a free, open, XML-based format for representing biochemical reaction networks. SBML is a software-independent language for describing models common to research in many areas of computational biology, including cell signaling pathways, metabolic pathways, gene regulation, and others. AVAILABILITY: The specification of SBML Level 1 is freely available from http://www.sbml.org/

BACKGROUND: Low muscle mass and weakness are common and potentially disabling in older adults, but in order to become recognized as a clinical condition, criteria for diagnosis should be based on clinically relevant thresholds and independently validated. The Foundation for the National Institutes of Health Biomarkers Consortium Sarcopenia Project used an evidence-based approach to develop these criteria. Initial findings were presented at a conference in May 2012, which generated recommendations that guided additional analyses to determine final recommended criteria. Details of the Project and its findings are presented in four accompanying manuscripts. METHODS: The Foundation for the National Institutes of Health Sarcopenia Project used data from nine sources of community-dwelling older persons: Age, Gene/Environment Susceptibility-Reykjavik Study, Boston Puerto Rican Health Study, a series of six clinical trials, Framingham Heart Study, Health, Aging, and Body Composition, Invecchiare in Chianti, Osteoporotic Fractures in Men Study, Rancho Bernardo Study, and Study of Osteoporotic Fractures. Feedback from conference attendees was obtained via surveys and breakout groups. RESULTS: The pooled sample included 26,625 participants (57% women, mean age in men 75.2 [±6.1 SD] and in women 78.6 [±5.9] years). Conference attendees emphasized the importance of evaluating the influence of body mass on cutpoints. Based on the analyses presented in this series, the final recommended cutpoints for weakness are grip strength <26kg for men and <16kg for women, and for low lean mass, appendicular lean mass adjusted for body mass index <0.789 for men and <0.512 for women. CONCLUSIONS: These evidence-based cutpoints, based on a large and diverse population, may help identify participants for clinical trials and should be evaluated among populations with high rates of functional limitations.

BACKGROUND: To assess potentially elevated cardiovascular risk related to new antihyperglycemic drugs in patients with type 2 diabetes, regulatory agencies require a comprehensive evaluation of the cardiovascular safety profile of new antidiabetic therapies. We assessed cardiovascular outcomes with alogliptin, a new inhibitor of dipeptidyl peptidase 4 (DPP-4), as compared with placebo in patients with type 2 diabetes who had had a recent acute coronary syndrome. METHODS: We randomly assigned patients with type 2 diabetes and either an acute myocardial infarction or unstable angina requiring hospitalization within the previous 15 to 90 days to receive alogliptin or placebo in addition to existing antihyperglycemic and cardiovascular drug therapy. The study design was a double-blind, noninferiority trial with a prespecified noninferiority margin of 1.3 for the hazard ratio for the primary end point of a composite of death from cardiovascular causes, nonfatal myocardial infarction, or nonfatal stroke. RESULTS: A total of 5380 patients underwent randomization and were followed for up to 40 months (median, 18 months). A primary end-point event occurred in 305 patients assigned to alogliptin (11.3%) and in 316 patients assigned to placebo (11.8%) (hazard ratio, 0.96; upper boundary of the one-sided repeated confidence interval, 1.16; P<0.001 for noninferiority). Glycated hemoglobin levels were significantly lower with alogliptin than with placebo (mean difference, -0.36 percentage points; P<0.001). Incidences of hypoglycemia, cancer, pancreatitis, and initiation of dialysis were similar with alogliptin and placebo. CONCLUSIONS: Among patients with type 2 diabetes who had had a recent acute coronary syndrome, the rates of major adverse cardiovascular events were not increased with the DPP-4 inhibitor alogliptin as compared with placebo. (Funded by Takeda Development Center Americas; EXAMINE ClinicalTrials.gov number, NCT00968708.).

The worldwide incidence of bone disorders and conditions has trended steeply upward and is expected to double by 2020, especially in populations where aging is coupled with increased obesity and poor physical activity. Engineered bone tissue has been viewed as a potential alternative to the conventional use of bone grafts, due to their limitless supply and no disease transmission. However, bone tissue engineering practices have not proceeded to clinical practice due to several limitations or challenges. Bone tissue engineering aims to induce new functional bone regeneration via the synergistic combination of biomaterials, cells, and factor therapy. In this review, we discuss the fundamentals of bone tissue engineering, highlighting the current state of this field. Further, we review the recent advances of biomaterial and cell-based research, as well as approaches used to enhance bone regeneration. Specifically, we discuss widely investigated biomaterial scaffolds, micro- and nano-structural properties of these scaffolds, and the incorporation of biomimetic properties and/or growth factors. In addition, we examine various cellular approaches, including the use of mesenchymal stem cells (MSCs), embryonic stem cells (ESCs), adult stem cells, induced pluripotent stem cells (iPSCs), and platelet-rich plasma (PRP), and their clinical application strengths and limitations. We conclude by overviewing the challenges that face the bone tissue engineering field, such as the lack of sufficient vascularization at the defect site, and the research aimed at functional bone tissue engineering. These challenges will drive future research in the field.

Abstract Cancer-associated fibroblasts (CAF) are major players in the progression and drug resistance of pancreatic ductal adenocarcinoma (PDAC). CAFs constitute a diverse cell population consisting of several recently described subtypes, although the extent of CAF heterogeneity has remained undefined. Here we use single-cell RNA sequencing to thoroughly characterize the neoplastic and tumor microenvironment content of human and mouse PDAC tumors. We corroborate the presence of myofibroblastic CAFs and inflammatory CAFs and define their unique gene signatures in vivo. Moreover, we describe a new population of CAFs that express MHC class II and CD74, but do not express classic costimulatory molecules. We term this cell population “antigen-presenting CAFs” and find that they activate CD4+ T cells in an antigen-specific fashion in a model system, confirming their putative immune-modulatory capacity. Our cross-species analysis paves the way for investigating distinct functions of CAF subtypes in PDAC immunity and progression. Significance: Appreciating the full spectrum of fibroblast heterogeneity in pancreatic ductal adenocarcinoma is crucial to developing therapies that specifically target tumor-promoting CAFs. This work identifies MHC class II–expressing CAFs with a capacity to present antigens to CD4+ T cells, and potentially to modulate the immune response in pancreatic tumors. See related commentary by Belle and DeNardo, p. 1001. This article is highlighted in the In This Issue feature, p. 983

Utilization of polymers as biomaterials has greatly impacted the advancement of modern medicine. Specifically, polymeric biomaterials that are biodegradable provide the significant advantage of being able to be broken down and removed after they have served their function. Applications are wide ranging with degradable polymers being used clinically as surgical sutures and implants. In order to fit functional demand, materials with desired physical, chemical, biological, biomechanical and degradation properties must be selected. Fortunately, a wide range of natural and synthetic degradable polymers has been investigated for biomedical applications with novel materials constantly being developed to meet new challenges. This review summarizes the most recent advances in the field over the past 4 years, specifically highlighting new and interesting discoveries in tissue engineering and drug delivery applications.

OBJECTIVE: To issue a recommendation on the types and amounts of physical activity needed to improve and maintain health in older adults. PARTICIPANTS: A panel of scientists with expertise in public health, behavioral science, epidemiology, exercise science, medicine, and gerontology. EVIDENCE: The expert panel reviewed existing consensus statements and relevant evidence from primary research articles and reviews of the literature. PROCESS: After drafting a recommendation for the older adult population and reviewing drafts of the Updated Recommendation from the American College of Sports Medicine (ACSM) and the American Heart Association (AHA) for Adults, the panel issued a final recommendation on physical activity for older adults. SUMMARY: The recommendation for older adults is similar to the updated ACSM/AHA recommendation for adults, but has several important differences including: the recommended intensity of aerobic activity takes into account the older adult's aerobic fitness; activities that maintain or increase flexibility are recommended; and balance exercises are recommended for older adults at risk of falls. In addition, older adults should have an activity plan for achieving recommended physical activity that integrates preventive and therapeutic recommendations. The promotion of physical activity in older adults should emphasize moderate-intensity aerobic activity, muscle-strengthening activity, reducing sedentary behavior, and risk management.

BACKGROUND: Sex and gender differences are often overlooked in research design, study implementation and scientific reporting, as well as in general science communication. This oversight limits the generalizability of research findings and their applicability to clinical practice, in particular for women but also for men. This article describes the rationale for an international set of guidelines to encourage a more systematic approach to the reporting of sex and gender in research across disciplines. METHODS: A panel of 13 experts representing nine countries developed the guidelines through a series of teleconferences, conference presentations and a 2-day workshop. An internet survey of 716 journal editors, scientists and other members of the international publishing community was conducted as well as a literature search on sex and gender policies in scientific publishing. RESULTS: The Sex and Gender Equity in Research (SAGER) guidelines are a comprehensive procedure for reporting of sex and gender information in study design, data analyses, results and interpretation of findings. CONCLUSIONS: The SAGER guidelines are designed primarily to guide authors in preparing their manuscripts, but they are also useful for editors, as gatekeepers of science, to integrate assessment of sex and gender into all manuscripts as an integral part of the editorial process.

Endospores of Bacillus spp., especially Bacillus subtilis, have served as experimental models for exploring the molecular mechanisms underlying the incredible longevity of spores and their resistance to environmental insults. In this review we summarize the molecular laboratory model of spore resistance mechanisms and attempt to use the model as a basis for exploration of the resistance of spores to environmental extremes both on Earth and during postulated interplanetary transfer through space as a result of natural impact processes.

Many intracellular pathogens infect a broad range of host tissues, but the importance of T cells for immunity in these sites is unclear because most of our understanding of antimicrobial T cell responses comes from analyses of lymphoid tissue. Here, we show that in response to viral or bacterial infection, antigen-specific CD8 T cells migrated to nonlymphoid tissues and were present as long-lived memory cells. Strikingly, CD8 memory T cells isolated from nonlymphoid tissues exhibited effector levels of lytic activity directly ex vivo, in contrast to their splenic counterparts. These results point to the existence of a population of extralymphoid effector memory T cells poised for immediate response to infection.

Osteoporosis is a disorder in which loss of bone strength leads to fragility fractures. This review examines the fundamental pathogenetic mechanisms underlying this disorder, which include: (a) failure to achieve a skeleton of optimal strength during growth and development; (b) excessive bone resorption resulting in loss of bone mass and disruption of architecture; and (c) failure to replace lost bone due to defects in bone formation. Estrogen deficiency is known to play a critical role in the development of osteoporosis, while calcium and vitamin D deficiencies and secondary hyperparathyroidism also contribute. There are multiple mechanisms underlying the regulation of bone remodeling, and these involve not only the osteoblastic and osteoclastic cell lineages but also other marrow cells, in addition to the interaction of systemic hormones, local cytokines, growth factors, and transcription factors. Polymorphisms of a large number of genes have been associated with differences in bone mass and fragility. It is now possible to diagnose osteoporosis, assess fracture risk, and reduce that risk with antiresorptive or other available therapies. However, new and more effective approaches are likely to emerge from a better understanding of the regulators of bone cell function.

Categorization of the Ehlers-Danlos syndromes began in the late 1960s and was formalized in the Berlin nosology. Over time, it became apparent that the diagnostic criteria established and published in 1988 did not discriminate adequately between the different types of Ehlers-Danlos syndromes or between Ehlers-Danlos syndromes and other phenotypically related conditions. In addition, elucidation of the molecular basis of several Ehlers-Danlos syndromes has added a new dimension to the characterization of this group of disorders. We propose a revision of the classification of the Ehlers-Danlos syndromes based primarily on the cause of each type. Major and minor diagnostic criteria have been defined for each type and complemented whenever possible with laboratory findings. This simplified classification will facilitate an accurate diagnosis of the Ehlers-Danlos syndromes and contribute to the delineation of phenotypically related disorders.

People with severe mental illness (SMI) - schizophrenia, bipolar disorder and major depressive disorder - appear at risk for cardiovascular disease (CVD), but a comprehensive meta-analysis is lacking. We conducted a large-scale meta-analysis assessing the prevalence and incidence of CVD; coronary heart disease; stroke, transient ischemic attack or cerebrovascular disease; congestive heart failure; peripheral vascular disease; and CVD-related death in SMI patients (N=3,211,768) versus controls (N=113,383,368) (92 studies). The pooled CVD prevalence in SMI patients (mean age 50 years) was 9.9% (95% CI: 7.4-13.3). Adjusting for a median of seven confounders, patients had significantly higher odds of CVD versus controls in cross-sectional studies (odds ratio, OR=1.53, 95% CI: 1.27-1.83; 11 studies), and higher odds of coronary heart disease (OR=1.51, 95% CI: 1.47-1.55) and cerebrovascular disease (OR=1.42, 95% CI: 1.21-1.66). People with major depressive disorder were at increased risk for coronary heart disease, while those with schizophrenia were at increased risk for coronary heart disease, cerebrovascular disease and congestive heart failure. Cumulative CVD incidence in SMI patients was 3.6% (95% CI: 2.7-5.3) during a median follow-up of 8.4 years (range 1.8-30.0). Adjusting for a median of six confounders, SMI patients had significantly higher CVD incidence than controls in longitudinal studies (hazard ratio, HR=1.78, 95% CI: 1.60-1.98; 31 studies). The incidence was also higher for coronary heart disease (HR=1.54, 95% CI: 1.30-1.82), cerebrovascular disease (HR=1.64, 95% CI: 1.26-2.14), congestive heart failure (HR=2.10, 95% CI: 1.64-2.70), and CVD-related death (HR=1.85, 95% CI: 1.53-2.24). People with major depressive disorder, bipolar disorder and schizophrenia were all at increased risk of CVD-related death versus controls. CVD incidence increased with antipsychotic use (p=0.008), higher body mass index (p=0.008) and higher baseline CVD prevalence (p=0.03) in patients vs. CONTROLS: Moreover, CVD prevalence (p=0.007), but not CVD incidence (p=0.21), increased in more recently conducted studies. This large-scale meta-analysis confirms that SMI patients have significantly increased risk of CVD and CVD-related mortality, and that elevated body mass index, antipsychotic use, and CVD screening and management require urgent clinical attention.

OBJECTIVES: To describe the changes in activities of daily living (ADL) function occurring before and after hospital admission in older people hospitalized with medical illness and to assess the effect of age on loss of ADL function. DESIGN: Prospective observational study. SETTING: The general medical service of two hospitals. PARTICIPANTS: Two thousand two hundred ninety-three patients aged 70 and older (mean age 80, 64% women, 24% nonwhite). MEASUREMENTS: At the time of hospital admission, patients or their surrogates were interviewed about their independence in five ADLs (bathing, dressing, eating, transferring, and toileting) 2 weeks before admission (baseline) and at admission. Subjects were interviewed about ADL function at discharge. Outcome measures included functional decline between baseline and discharge and functional changes between baseline and admission and between admission and discharge. RESULTS: Thirty-five percent of patients declined in ADL function between baseline and discharge. This included the 23% of patients who declined between baseline and admission and failed to recover to baseline function between admission and discharge and the 12% of patients who did not decline between baseline and admission but declined between hospital admission and discharge. Twenty percent of patients declined between baseline and admission but recovered to baseline function between admission and discharge. The frequency of ADL decline between baseline and discharge varied markedly with age (23%, 28%, 38%, 50%, and 63% in patients aged 70-74, 75-79, 80-84, 85-89, and > or =90, respectively, P <.001). After adjustment for potential confounders, age was not associated with ADL decline before hospitalization (odds ratio (OR) for patients aged > or =90 compared with patients aged 70-74 = 1.26, 95% confidence interval (CI) = 0.88-1.82). In contrast, age was associated with the failure to recover ADL function during hospitalization in patients who declined before admission (OR for patients aged > or =90 compared with patients aged 70-74 = 2.09, 95% CI = 1.20-3.65) and with new losses of ADL function during hospitalization in patients who did not decline before admission (OR for patients aged > or =90 compared with patients aged 70-74 = 3.43, 95% CI = 1.92-6.12). CONCLUSION: Many hospitalized older people are discharged with ADL function that is worse than their baseline function. The oldest patients are at particularly high risk of poor functional outcomes because they are less likely to recover ADL function lost before admission and more likely to develop new functional deficits during hospitalization

BACKGROUND: We conducted a double-blind, randomized, placebo-controlled trial to determine the effects of oral cyclophosphamide on lung function and health-related symptoms in patients with evidence of active alveolitis and scleroderma-related interstitial lung disease. METHODS: At 13 clinical centers throughout the United States, we enrolled 158 patients with scleroderma, restrictive lung physiology, dyspnea, and evidence of inflammatory interstitial lung disease on examination of bronchoalveolar-lavage fluid, thoracic high-resolution computed tomography, or both. Patients received oral cyclophosphamide (< or =2 mg per kilogram of body weight per day) or matching placebo for one year and were followed for an additional year. Pulmonary function was assessed every three months during the first year, and the primary end point was the forced vital capacity (FVC, expressed as a percentage of the predicted value) at 12 months, after adjustment for the baseline FVC. RESULTS: Of 158 patients, 145 completed at least six months of treatment and were included in the analysis. The mean absolute difference in adjusted 12-month FVC percent predicted between the cyclophosphamide and placebo groups was 2.53 percent (95 percent confidence interval, 0.28 to 4.79 percent), favoring cyclophosphamide (P<0.03). There were also treatment-related differences in physiological and symptom outcomes, and the difference in FVC was maintained at 24 months. There was a greater frequency of adverse events in the cyclophosphamide group, but the difference between the two groups in the number of serious adverse events was not significant. CONCLUSIONS: One year of oral cyclophosphamide in patients with symptomatic scleroderma-related interstitial lung disease had a significant but modest beneficial effect on lung function, dyspnea, thickening of the skin, and the health-related quality of life. The effects on lung function were maintained through the 24 months of the study.

Mice bearing subcutaneous EMT-6 mammary carcinomas received a single intravenous injection of 1.9 nm diameter gold particles (up to 2.7 g Au/kg body weight), which elevated concentrations of gold to 7 mg Au/g in tumours. Tumour-to-normal-tissue gold concentration ratios remained approximately 8:1 during several minutes of 250 kVp x-ray therapy. One-year survival was 86% versus 20% with x-rays alone and 0% with gold alone. The increase in tumours safely ablated was dependent on the amount of gold injected. The gold nanoparticles were apparently non-toxic to mice and were largely cleared from the body through the kidneys. This novel use of small gold nanoparticles permitted achievement of the high metal content in tumours necessary for significant high-Z radioenhancement.

BACKGROUND: Osteoporosis is a major public health problem of largely unknown cause. Loss-of-function mutations in the gene for low-density lipoprotein receptor-related protein 5 (LRP5), which acts in the Wnt signaling pathway, have been shown to cause osteoporosis-pseudoglioma. METHODS: We performed genetic and biochemical analyses of a kindred with an autosomal dominant syndrome characterized by high bone density, a wide and deep mandible, and torus palatinus. RESULTS: Genetic analysis revealed linkage of the syndrome to chromosome 11q12-13 (odds of linkage, >1 million to 1), an interval that contains LRP5. Affected members of the kindred had a mutation in this gene, with valine substituted for glycine at codon 171 (LRP5V171). This mutation segregated with the trait in the family and was absent in control subjects. The normal glycine lies in a so-called propeller motif that is highly conserved from fruit flies to humans. Markers of bone resorption were normal in the affected subjects, whereas markers of bone formation such as osteocalcin were markedly elevated. Levels of fibronectin, a known target of signaling by Wnt, a developmental protein, were also elevated. In vitro studies showed that the normal inhibition of Wnt signaling by another protein, Dickkopf-1 (Dkk-1), was defective in the presence of LRP5V171 and that this resulted in increased signaling due to unopposed Wnt activity. CONCLUSIONS: The LRP5V171 mutation causes high bone density, with a thickened mandible and torus palatinus, by impairing the action of a normal antagonist of the Wnt pathway and thus increasing Wnt signaling. These findings demonstrate the role of altered LRP5 function in high bone mass and point to Dkk as a potential target for the prevention or treatment of osteoporosis.

Abstract The cell envelope of Salmonella typhimurium has been separated into cytoplasmic and outer membrane fractions of relatively high purity by a new method based on isopycnic sucrose density gradient centrifugation of the total membrane fraction obtained by lysis of lysozyme-EDTA spheroplasts. The outer membrane fraction contained approximately 60% of the protein of the total membranes, 50% of the phospholipid, and 90% of the lipopolysaccharide. The cytoplasmic and outer membrane fractions differed markedly in over-all protein composition as determined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate as well as in specific enzyme activities. Total cytochromes, DPNH oxidase, succinate dehydrogenase, d-lactate dehydrogenase, and Enzyme II of the α-methylglucoside phosphotransferase system were recovered essentially exclusively in cytoplasmic membrane; the specific activities of these in the outer membrane were 1 to 5% of the values obtained for cytoplasmic membrane fractions. In contrast, a phospholipase activity tentatively identified as a mixture of phospholipase A and lysophospholipase, appeared to be localized primarily in the outer membrane. UDP-sugar hydrolase, ribonuclease I and endonuclease I activities were associated with both membrane fractions. Preliminary analysis of the phospholipid composition of the isolated fractions also showed significant quantitative differences in the relative distribution of the major glycerophosphatides. The ratios of phosphatidylglycerol to phosphatidylethanolamine and of cardiolipin to phosphatidylethanolamine in the outer membrane were approximately one-half and one-quarter those of the cytoplasmic membrane.