Comparison of N-Glycopeptide to Released N-Glycan Abundances and the Influence of Glycopeptide Mass and Charge States on N-Linked Glycosylation of IgG Antibodies.

We report the comparison of mass-spectral-based abundances of tryptic glycopeptides to fluorescence abundances of released labeled glycans and the effects of mass and charge state and in-source fragmentation on glycopeptide abundances. The primary glycoforms derived from Rituximab, NISTmAb, Evolocumab, and Infliximab were high-mannose and biantennary complex galactosylated and fucosylated N-glycans. Except for Evolocumab, in-source ions derived from the loss of HexNAc or HexNAc-Hex sugars are prominent for other therapeutic IgGs. After excluding in-source fragmentation of glycopeptide ions from the results, a linear correlation was observed between fluorescently labeled N-glycan and glycopeptide abundances over a dynamic range of 500. Different charge states of human IgG-derived glycopeptides containing a wider variety of abundant attached glycans were also investigated to examine the effects of the charge state on ion abundances. These revealed a linear dependence of glycopeptide abundance on the mass of the glycan with higher charge states favoring higher-mass glycans. Findings indicate that the mass spectrometry-based bottom-up approach can provide results as accurate as those of glycan release studies while revealing the origin of each attached glycan. These site-specific relative abundances are conveniently displayed and compared using previously described glycopeptide abundance distribution spectra "GADS" representations. Mass spectrometry data are available from the MAssIVE repository (MSV000093562).

Economic Evaluation of Early Psychosis Interventions From A Canadian Perspective.

BACKGROUND: Compared to treatment as usual (TAU), early psychosis intervention programs (EPI) have been shown to reduce mortality, hospitalizations and days of assisted living while improving employment status. AIMS: The study aim was to conduct a cost-benefit analysis (CBA) and a cost-effectiveness analysis (CEA) to compare EPI and TAU in Canada. METHODS: A decision-analytic model was used to estimate the 5-year costs and benefits of treating patients with a first episode of psychosis with EPI or TAU. EPI benefits were derived from randomized controlled trials (RCTs) and Canadian administrative data. The cost of EPI was based on a published survey of 52 EPI centers in Canada while hospitalizations, employment and days of assisted living were valued using Canadian unit costs. The outcomes of the CBA and CEA were expressed in terms of net benefit (NB) and incremental cost per life year gained (LYG), respectively. Scenario analyses were conducted to examine the impact of key assumptions. Costs are reported in 2019 Canadian dollars. RESULTS: Base case results indicated that EPI had a NB of $85,441 (95% CI: $41,140; $126,386) compared to TAU while the incremental cost per LYG was $26,366 (95% CI: EPI dominates TAU (less costs, more life years); $102,269). In all sensitivity analyses the NB of EPI remained positive and the incremental cost per LYG was less than $50,000. CONCLUSIONS: In addition to EPI demonstrated clinical benefits, our results suggest that large-scale implementation of EPI in Canada would be desirable from an economic point of view .

Comparison between intra-articular infiltrations of placebo, steroids, hyaluronic and PRP for knee osteoarthritis: a Bayesian network meta-analysis.

INTRODUCTION: Regarding the efficacy of intra-articular injections of platelet-rich plasma, hyaluronic acid and corticosteroids, current evidence is controversial. The superiority of one technique over another is questioned and debates are ongoing. The purpose of the present study was to compare and investigate the efficacy of these intra-articular infiltrations in patients with knee osteoarthritis (OA). A Bayesian network meta-analysis of randomized clinical trials (RCTs) was conducted comparing patient outcomes at 3, 6 and 12-months of follow-up. MATERIALS AND METHODS: This Bayesian network meta-analysis was conducted according to the PRISMA extension statement for reporting systematic reviews incorporating network meta-analyses of health care interventions. All the RCTs comparing the outcomes of two or more intra-articular infiltrations of interest for knee OA were considered for inclusion. The outcomes of interest were the WOMAC and VAS scores. The network meta-analyses were performed using the STATA routine for Bayesian hierarchical random-effects models. RESULTS: Data from 30 RCTs (3463 patients) were collected. At 3-months follow-up, PRP showed the best WOMAC scores, followed by the Placebo, CCS and HA. At 6-months follow-up, PRP showed the best WOMAC scores, followed by HA, CCS and Placebo. At 12-months follow-up, PRP showed the best WOMAC scores, followed by the Placebo, HA and CCS. At 3-months follow-up, the PRP showed the best VAS scores, followed by CCS, HA and Placebo. At 6-months follow-up, PRP showed the best VAS scores, followed by CCS, Placebo and HA. At 12-months follow-up, the PRP showed the best VAS scores, followed by CCS, Placebo and HA. CONCLUSION: Intra-articular injections of PRP demonstrated the best overall outcome compared to steroids, hyaluronic acid and placebo for patients with knee osteoarthrosis at 3, 6 and 12-months follow-up. Among CCS, hyaluronic acid and placebo, no discrepancies were detected. LEVEL OF EVIDENCE: I, Bayesian network meta-analysis of RCTs.

Mass Spectral Library of Acylcarnitines Derived from Human Urine.

We describe the creation of a mass spectral library of acylcarnitines and conjugated acylcarnitines from the LC-MS/MS analysis of six NIST urine reference materials. To recognize acylcarnitines, we conducted in-depth analyses of fragmentation patterns of acylcarnitines and developed a set of rules, derived from spectra in the NIST17 Tandem MS Library and those identified in urine, using the newly developed hybrid search method. Acylcarnitine tandem spectra were annotated with fragments from carnitine and acyl moieties as well as neutral loss peaks from precursors. Consensus spectra were derived from spectra having similar retention time, fragmentation pattern, and the same precursor m/z and collision energy. The library contains 157 different precursor masses, 586 unique acylcarnitines, and 4 332 acylcarnitine consensus spectra. Furthermore, from spectra that partially satisfied the fragmentation rules of acylcarnitines, we identified 125 conjugated acylcarnitines represented by 987 consensus spectra, which appear to originate from Phase II biotransformation reactions. To our knowledge, this is the first report of conjugated acylcarnitines. The mass spectra provided by this work may be useful for clinical screening of acylcarnitines as well as for studying relationships among fragmentation patterns, collision energies, structures, and retention times of acylcarnitines. Further, these methods are extensible to other classes of metabolites.

Evaluation of geriatrics education at a Chinese University: A survey of attitudes and knowledge among undergraduate medical students.

Despite widespread reforms in medical education across China, nationally there has been no mandate or movement toward systemically incorporating geriatrics into curricula. To what degree medical students are trained and have exposure to geriatric topics remains unclear. We surveyed 190 medical students during their final year of medical school at a Chinese medical university, graduating from reformed and also traditional curricula. The survey was comprised of a subjective assessment of attitudes and reported knowledge, as well as an objective assessment of knowledge via a multiple choice test. Student attitudes were favorable toward geriatrics, with 91% supporting the addition of specialized clinical experiences to the curriculum. Students generally reported low exposure to geriatrics, with no statistically significant differences between reform and traditional curricula. There was a statistically significant difference in performance on the multiple choice test between curricula but at a degree unlikely to be practically significant. Students had very favorable attitudes toward geriatrics as a field and specialty; however scored poorly on competency exams, with the lowest performance around diagnosis and treatment of specific geriatric conditions. Our results suggest that there is a need and desire for increased geriatric-oriented learning at Chinese medical schools.

Hybrid Search: A Method for Identifying Metabolites Absent from Tandem Mass Spectrometry Libraries.

Metabolomics has a critical need for better tools for mass spectral identification. Common metabolites may be identified by searching libraries of tandem mass spectra, which offers important advantages over other approaches to identification. But tandem libraries are not nearly complete enough to represent the full molecular diversity present in complex biological samples. We present a novel hybrid search method that can help identify metabolites not in the library by similarity to compounds that are. We call it "hybrid" searching because it combines conventional, direct peak matching with the logical equivalent of neutral-loss matching. A successful hybrid search requires the library to contain "cognates" of the unknown: similar compounds with a structural difference confined to a single region of the molecule, that does not substantially alter its fragmentation behavior. We demonstrate that the hybrid search is highly likely to find similar compounds under such circumstances.

Correction to: Clinical diversity of invasive cryptococcosis in AIDS patients from central China: report of two cases with review of literature.

After publication of the original article [1], we were notified that Figs. 1 and 2 has been misplaced. Hence, the position of the two pictures should be reversed.

Clinical diversity of invasive cryptococcosis in AIDS patients from central China: report of two cases with review of literature.

BACKGROUND: Although antiretroviral therapy (ART) has greatly improved the prognosis of acquired immunodeficiency syndrome (AIDS) patients globally, opportunistic infections (OIs) are still common in Chinese AIDS patients, especially cryptococcosis. CASE PRESENTATION: We described here two Chinese AIDS patients with cryptococcal infections. Case one was a fifty-year-old male. At admission, he was conscious and oriented, with papulonodular and umbilicated skin lesions, some with ulceration and central necrosis resembling molluscum contagiosum. The overall impression reminded us of talaromycosis: we therefore initiated empirical treatment with amphotericin B, even though the case history of this patient did not support such a diagnosis. On the second day of infusion, the patient complained of intermittent headache, but the brain CT revealed no abnormalities. On the third day, a lumbar puncture was performed. The cerebral spinal fluid (CSF) was turbid, with slightly increased pressure. India ink staining was positive, but the cryptococcus antigen latex agglutination test (CrAgLAT: IMMY, USA) was negative. Two days later, the blood culture showed a growth of Cryptococcus neoformans, and the same result came from the skin culture. We added fluconazole to the patient's treatment, but unfortunately, he died three days later. Case two was a sixty-four-year-old female patient with mild fever, productive cough, dyspnea upon movement, and swelling in both lower limbs. The patient was empirically put on cotrimoxazole per os and moxifloxacin by infusion. A bronchofibroscopy was conducted with a fungal culture, showing growth of Cryptococcus laurentii colonies. Amphotericin B was started thereafter but discontinued three days later in favor of fluconazole 400 mg/d due to worsening renal function. The patient became afebrile after 72 h of treatment with considerable improvement of other comorbidities and was finally discharged with continuing oral antifungal therapy. CONCLUSIONS: Our cases illustrate that cryptococcal disease is an important consideration when treating immunocompromised individuals such as AIDS patients. Life threatening meningitis or meningoencephalitis caused by C. neoformansmay still common in these populations and can vary greatly in clinical presentations, especially with regard to skin lesions. Pulmonary cryptococcosis caused by C. laurentii is rare, but should also be considered in certain contexts. Guidelines for its earlier diagnosis, treatment and prophylaxis are needed.

Effects of Work-Related Stressors and Mindfulness on Mental and Physical Health Among Australian Nurses and Healthcare Workers.

PURPOSE: To examine the relative impact of work-related stressors and the personal resource of mindfulness on employees' mental and physical health. DESIGN: A cross-sectional survey design with nursing and healthcare workers in Victoria, Australia. METHODS: Data were collected from 702 respondents. Mean scores for work-related stressors and employee mental and physical health were compared with population norms. We used hierarchical linear regressions to examine the relative impact of demographics, work-related stressors, and mindfulness on employee mental and physical health. FINDINGS: Employees in this sample reported higher levels of work-related stress and poorer mental health compared to available norms, while their levels of physical health were within the normal range. Regression analyses showed that work-related stressors were important predictors of employee mental health, but mindfulness was the stronger predictor. There was a slightly stronger relationship between employee physical health and work-related stress compared to mindfulness. Furthermore, being younger and employed in a non-nursing role were associated with better physical health. CLINICAL RELEVANCE: Encouraging mindfulness as a health behavior practice among nurses and other healthcare workers could improve employee well-being and potentially enable them to more effectively fulfill the requirements of their demanding roles.

The impact of perceived workload on nurse satisfaction with work-life balance and intention to leave the occupation.

BACKGROUND AND AIM: In the drive to make the health sector more economically efficient and effective, what is potentially being lost is the need to look after the well-being of those who work within this profession. Nurses are the largest group in the health sector workforce and the frontline of patient care. Workload perceptions are known to be impacting nurses' well-being and are becoming a critical concern for the retention of this workforce. In response, this study aims to examine the relationships among perceived workload, satisfaction with work-life balance (an indicator of well-being), and intention to leave the occupation. Additionally, high involvement work practices (HIWPs) are examined as a form of organisational support that may buffer the negative impact of perceived workload on nurses' well-being and intention to leave the occupation. METHOD: A 2016 online survey of the nursing profession in Australia yielded 2984 responses. We assessed the impact of perceived workload on nurses' well-being and intention to leave the occupation, and the role of HIWPs in ameliorating the negative impact of perceived workload. RESULTS AND CONCLUSION: Our results show that perceived workload is associated with increasing intention to leave the occupation and is mediated by nurses' satisfaction with work-life balance. Where organisational support is provided through HIWPs, this can mitigate such intentions. These aspects are within the control of those who manage this workforce and should be central to human resource management strategies in the health care sector.

GEneSTATION 1.0: a synthetic resource of diverse evolutionary and functional genomic data for studying the evolution of pregnancy-associated tissues and phenotypes.

Mammalian gestation and pregnancy are fast evolving processes that involve the interaction of the fetal, maternal and paternal genomes. Version 1.0 of the GEneSTATION database (http://genestation.org) integrates diverse types of omics data across mammals to advance understanding of the genetic basis of gestation and pregnancy-associated phenotypes and to accelerate the translation of discoveries from model organisms to humans. GEneSTATION is built using tools from the Generic Model Organism Database project, including the biology-aware database CHADO, new tools for rapid data integration, and algorithms that streamline synthesis and user access. GEneSTATION contains curated life history information on pregnancy and reproduction from 23 high-quality mammalian genomes. For every human gene, GEneSTATION contains diverse evolutionary (e.g. gene age, population genetic and molecular evolutionary statistics), organismal (e.g. tissue-specific gene and protein expression, differential gene expression, disease phenotype), and molecular data types (e.g. Gene Ontology Annotation, protein interactions), as well as links to many general (e.g. Entrez, PubMed) and pregnancy disease-specific (e.g. PTBgene, dbPTB) databases. By facilitating the synthesis of diverse functional and evolutionary data in pregnancy-associated tissues and phenotypes and enabling their quick, intuitive, accurate and customized meta-analysis, GEneSTATION provides a novel platform for comprehensive investigation of the function and evolution of mammalian pregnancy.

Postincident Support for Healthcare Workers Experiencing Occupational Violence and Aggression.

PURPOSE: To investigate the relative contributions of workplace type, occupational violence and aggression (OVA) strategies and interventions along with perceptions of the occupational health and safety (OHS) environment on the likelihood of receiving postincident support following the experience of OVA. DESIGN: We used a cross-sectional study design with an online survey to collect data from employees in nursing and midwifery in Victoria, Australia. METHODS: Survey data collected from 3,072 members of the Australian Nursing and Midwifery Federation (Victorian branch) were analyzed using logistic regression. FINDINGS: Of the 3,072 respondents who had experienced OVA in the preceding 12 months, 1,287 (42%) reported that they had received postincident support. Hierarchical logistic regression revealed that the OHS environment was the dominant factor that predicted the likelihood of workers receiving postincident support. Working in a positive OHS environment characterized by higher levels of leading indicators of OHS, prioritization of OHS, supervisor support for safety, and team psychological safety was the stronger predictor of postincident support. Being employed in a workplace that offered training in the management and prevention of OVA also increased the likelihood of receiving postincident support. CONCLUSIONS: While training in the management and prevention of OVA contributed to the likelihood of receiving postincident support, a greater emphasis on the OHS environment was more important in predicting the likelihood that workers received support. CLINICAL RELEVANCE: This study identifies workplace practices that facilitate the provision of postincident support for healthcare workers. Facilitating effective postincident support could improve outcomes for workers, their patients and workplaces, and society in general.

Twelve tips for using clinical cases to teach medical ethics.

The essential role of clinical case discussions in the teaching of medical ethics is well recognized. Based upon published literature and the authors' curricular development experience, the following 12 tips cover all major aspects of the case method for teaching clinical ethics and offer practical suggestions for designing and conducting case discussions.

Occupational Violence and Aggression Experienced by Nursing and Caring Professionals.

PURPOSE: To examine the extent and source of occupational violence and aggression (OVA) experienced by nursing and caring professionals. This study also examines the relative contributions of demographic characteristics and workplace and individual safety factors in predicting OVA. DESIGN: A cross-sectional study design with data collected using an online survey of employees in the nursing and caring professions in Victoria, Australia. METHODS: Survey data collected from 4,891 members of the Australian Nursing and Midwifery Federation (Victorian branch) were analyzed using logistic regression. FINDINGS: Sixty-seven percent of respondents reported experiencing OVA in the preceding 12 months, with nearly 20% experiencing OVA on a weekly or daily basis. The dominant sources of OVA were patients (79%) or relatives of patients (48%). Logistic regression analysis revealed that respondents working in public hospitals and aged care facilities were more likely to experience OVA, compared to those working in other workplaces. While higher levels of safety compliance reduced the likelihood of experiencing OVA, role overload and workplace safety factors such as prioritization of employee safety and leading indicators of occupational health and safety were stronger predictors. CONCLUSIONS: The likelihood of healthcare workers experiencing OVA varies across demographic and workplace characteristics. While some demographic characteristics and individual safety factors were significant predictors, our results suggest that a greater reduction in OVA could be achieved by improving workplace safety. CLINICAL RELEVANCE: The study's outcomes identify workforce segments that are most vulnerable to OVA. The study also highlights workplace safety factors such as the prioritization of employee safety that might assist in the reduction of OVA.

Medical ethics education in China: Lessons from three schools.

BACKGROUND: Ethics teaching is a relatively new area of medical education in China, with ethics curricula at different levels of development. This study examined ethics education at three medical schools in China to understand their curricular content, teaching and learning methods, forms of assessments, changes over time, and what changes are needed for further improvement. METHODS: We used student and faculty surveys to obtain information about the ethics courses' content, teaching methods, and revisions over time. The surveys also included five realistic cases and asked participants whether each would be appropriate to use for discussion in ethics courses. Students rated the cases on a scale and gave written comments. Finally, participants were asked to indicate how much they would agree with the statement that medical professionalism is about putting the interests of patients and society above one's own. RESULTS: There were both similarities and differences among these schools with regard to course topics, teaching and assessment methods, and course faculty compositions, suggesting their courses are at different levels of development. Areas of improvement for the schools' courses were identified based on this study's findings and available literature. A model of the evolution of medical ethics education in China was proposed to guide reform in medical ethics instruction in China. Analysis identified characteristics of appropriate cases and participants' attitudes toward the ideal of professionalism. DISCUSSION: We conclude that the development of medical ethics education in China is promising while much improvement is needed. In addition, ethics education is not confined to the walls of medical schools; the society at large can have significant influence on the formation of students' professional values.

Distinct subclassification of DRG neurons innervating the distal colon and glans penis/distal urethra based on the electrophysiological current signature.

Spinal sensory neurons innervating visceral and mucocutaneous tissues have unique microanatomic distribution, peripheral modality, and physiological, pharmacological, and biophysical characteristics compared with those neurons that innervate muscle and cutaneous tissues. In previous patch-clamp electrophysiological studies, we have demonstrated that small- and medium-diameter dorsal root ganglion (DRG) neurons can be subclassified on the basis of their patterns of voltage-activated currents (VAC). These VAC-based subclasses were highly consistent in their action potential characteristics, responses to algesic compounds, immunocytochemical expression patterns, and responses to thermal stimuli. For this study, we examined the VAC of neurons retrogradely traced from the distal colon and the glans penis/distal urethra in the adult male rat. The afferent population from the distal colon contained at least two previously characterized cell types observed in somatic tissues (types 5 and 8), as well as four novel cell types (types 15, 16, 17, and 18). In the glans penis/distal urethra, two previously described cell types (types 6 and 8) and three novel cell types (types 7, 14, and 15) were identified. Other characteristics, including action potential profiles, responses to algesic compounds (acetylcholine, capsaicin, ATP, and pH 5.0 solution), and neurochemistry (expression of substance P, CGRP, neurofilament, TRPV1, TRPV2, and isolectin B4 binding) were consistent for each VAC-defined subgroup. With identification of distinct DRG cell types that innervate the distal colon and glans penis/distal urethra, future in vitro studies related to the gastrointestinal and urogenital sensory function in normal as well as abnormal/pathological conditions may be benefitted.

Persistent modification of Nav1.9 following chronic exposure to insecticides and pyridostigmine bromide.

Many veterans of the 1991 Gulf War (GW) returned from that conflict with a widespread chronic pain affecting deep tissues. Recently, we have shown that a 60day exposure to the insecticides permethrin, chlorpyrifos, and pyridostigmine bromide (NTPB) had little influence on nociceptor action potential forming Nav1.8, but increased Kv7 mediated inhibitory currents 8weeks after treatment. Using the same exposure regimen, we used whole cell patch methods to examine whether the influences of NTPB could be observed on Nav1.9 expressed in muscle and vascular nociceptors. During a 60day exposure to NTPB, rats exhibited lowered muscle pain thresholds and increased rest periods, but these measures subsequently returned to normal levels. Eight and 12weeks after treatments ceased, DRG neurons were excised from the sensory ganglia. Whole cell patch studies revealed little change in voltage dependent activation and deactivation of Nav1.9, but significant increases in the amplitude of Nav1.9 were observed 8weeks after exposure. Cellular studies, at the 8week delay, revealed that NTPB also significantly prolonged action potential duration and afterhyperpolarization (22°C). Acute application of permethrin (10µM) also increased the amplitude of Nav1.9 in skin, muscle and vascular nociceptors. In conclusion, chronic exposure to Gulf War agents produced long term changes in the amplitude of Nav1.9 expressed in muscle and vascular nociceptors. The reported increases in Kv7 amplitude may have been an adaptive response to increased Nav1.9, and effectively suppressed behavioral pain measures in the post treatment period. Factors that alter the balance between Nav1.9 and Kv7 could release spontaneous discharge and produce chronic deep tissue pain.

Positive impact of integrating histology and physiology teaching at a medical school in China.

To modernize its stagnant, traditional curriculum and pedagogy, the Medical School of Wuhan University in China adopted (with modifications) the University of Chicago's medical curriculum model. The reform effort in basic sciences was integrating histology and physiology into one course, increasing the two subjects' connection to clinical medicine, and applying new pedagogies and assessment methods. This study assessed the results of the reform by comparing the attitudes and academic achievements of students in the reform curriculum (n = 41) and their traditional curriculum peers (n = 182). An attitude survey was conducted to obtain students' views of their respective histology and physiology instruction. Survey items covered lectures, laboratory teaching, case analyses and small-group case discussions, assessment of students, and overall quality of the courses and instruction. A knowledge test consisting of questions from three sources was given to measure students' mastery of topics that they had learned. Results showed that reform curriculum students were rather satisfied with their course and new teaching methods in most cases. When these students' attitudes were compared with those of their traditional curriculum peers, several significant differences favoring the reform were identified regarding physiology teaching. No other significant difference was found for physiology or histology teaching. Reform curriculum students outperformed their peers on four of five subcategories of the knowledge test questions. These findings support the benefits of integration and state-of-the-art teaching methods. Our study may offer lessons to medical schools in China and other countries whose medical education is in need of change.

Medical education reform in wuhan university, china: a preliminary report of an international collaboration.

BACKGROUND: In 2008 Wuhan University Medical School in China proposed to reform its curriculum by adapting the curriculum of the University of Chicago Medical School. DESCRIPTION: An assessment of Wuhan University Medical School's traditional curriculum conducted in 2009 informed the reform directions, which included course integration, use of clinical cases, improved relevance of basic sciences to clinical medicine, reduction of lecture time, increase in group and independent learning, and the use of formative assessments. Fifty student volunteers per year were chosen to participate in the reform, and the rest remained in the traditional curriculum. EVALUATION: A student survey was conducted in 2011 to evaluate the reform by comparing the attitudes of those in the reform and standard curricula. CONCLUSIONS: The reform met the needs of the school, was generally well received, improved satisfaction in reform participants, and had a positive impact on students. Areas needing improvement were also identified.