Broadening participation in community problem solving: a multidisciplinary model to support collaborative practice and research.
Over the last 40 years, thousands of communities-in the United States and internationally-have been working to broaden the involvement of people and organizations in addressing community-level problems related to health and other areas. Yet, in spite of this experience, many communities are having substantial difficulty achieving their collaborative objective, and many funders of community partnerships and participation initiatives are looking for ways to get more out of their investment. One of the reasons we are in this predicament is that the practitioners and researchers who are interested in community collaboration come from a variety of contexts, initiatives, and academic disciplines, and few of them have integrated their work with experiences or literatures beyond their own domain. In this article, we seek to overcome some of this fragmentation of effort by presenting a multidisciplinary model that lays out the pathways by which broadly participatory processes lead to more effective community problem solving and to improvements in community health. The model, which builds on a broad array of practical experience as well as conceptual and empirical work in multiple fields, is an outgrowth of a joint-learning work group that was organized to support nine communities in the Turning Point initiative. Following a detailed explication of the model, the article focuses on the implications of the model for research, practice, and policy. It describes how the model can help researchers answer the fundamental effectiveness and "how-to" questions related to community collaboration. In addition, the article explores differences between the model and current practice, suggesting strategies that can help the participants in, and funders of, community collaborations strengthen their efforts.


Analysis Of The Health Care System Of Pakistan: Lessons Learnt And Way Forward.
Pakistani health care system is in progress and since last year, Pakistan has tried to make much improvement in its health care delivery system and has brought out many reforms. A systematic search of national and international literature was looked from peerreviewed databases form MEDLINE, CINAHL, and PubMed. There is little strength in health care delivery system in Pakistan like making health policies, participating in Millennium Development Goals program, initiating vertical programs and introducing Public Private Partnership, improving human resource development and infrastructure by making Basic Health Unit and Rural Health Centres. However, these all programs are very limited in its scope and that is the reason that Pakistan's healthcare system is still not very efficient. There are numerous weaknesses like poor governance, lack of access and unequal resources, poor quality of Health Information Management System, corruption in health system, lack of monitoring in health policy and health planning and lack of trained staff. Pakistan is improving very slowly in the health sector for the last five decades as is evident by its health indicators and above mentioned strengths and weaknesses. Therefore, the Government needs to take strong initiatives to change the current health care system.


Antidepressants for people with epilepsy and depression.
Depressive disorders are the most common psychiatric comorbidity in people with epilepsy, affecting around one-third, with a significant negative impact on quality of life. There is concern that people may not be receiving appropriate treatment for their depression because of uncertainty regarding which antidepressant or class works best, and the perceived risk of exacerbating seizures. This review aimed to address these issues, and inform clinical practice and future research. This is an updated version of the original Cochrane Review published in Issue 12, 2014. To evaluate the efficacy and safety of antidepressants in treating depressive symptoms and the effect on seizure recurrence, in people with epilepsy and depression. For this update, we searched CRS Web, MEDLINE, SCOPUS, PsycINFO, and ClinicalTrials.gov (February 2021). We searched the World Health Organization Clinical Trials Registry in October 2019, but were unable to update it because it was inaccessible. There were no language restrictions. We included randomised controlled trials (RCTs) and prospective non-randomised studies of interventions (NRSIs), investigating children or adults with epilepsy, who were treated with an antidepressant and compared to placebo, comparative antidepressant, psychotherapy, or no treatment for depressive symptoms. DATA COLLECTION AND ANALYSIS: The primary outcomes were changes in depression scores (proportion with a greater than 50% improvement, mean difference, and proportion who achieved complete remission) and change in seizure frequency (mean difference, proportion with a seizure recurrence, or episode of status epilepticus). Secondary outcomes included the number of participants who withdrew from the study and reasons for withdrawal, quality of life, cognitive functioning, and adverse events. Two review authors independently extracted data for each included study. We then cross-checked the data extraction. We assessed risk of bias using the Cochrane tool for RCTs, and the ROBINS-I for NRSIs. We presented binary outcomes as risk ratios (RRs) with 95% confidence intervals (CIs) or 99% CIs for specific adverse events. We presented continuous outcomes as standardised mean differences (SMDs) with 95% CIs, and mean differences (MDs) with 95% CIs. MAIN RESULTS: We included 10 studies in the review (four RCTs and six NRSIs), with 626 participants with epilepsy and depression, examining the effects of antidepressants. One RCT was a multi-centre study comparing an antidepressant with cognitive behavioural therapy (CBT). The other three RCTs were single-centre studies comparing an antidepressant with an active control, placebo, or no treatment. The NRSIs reported on outcomes mainly in participants with focal epilepsy before and after treatment for depression with a selective serotonin reuptake inhibitor (SSRI); one NRSI compared SSRIs to CBT. We rated one RCT at low risk of bias, three RCTs at unclear risk of bias, and all six NRSIs at serious risk of bias. We were unable to conduct any meta-analysis of RCT data due to heterogeneity of treatment comparisons. We judged the certainty of evidence to be moderate to very low across comparisons, because single studies contributed limited outcome data, and because of risk of bias, particularly for NRSIs, which did not adjust for confounding variables. More than 50% improvement in depressive symptoms ranged from 43% to 82% in RCTs, and from 24% to 97% in NRSIs, depending on the antidepressant given. Venlafaxine improved depressive symptoms by more than 50% compared to no treatment (mean difference (MD) -7.59 (95% confidence interval (CI) -11.52 to -3.66; 1 study, 64 participants; low-certainty evidence); the results between other comparisons were inconclusive. Two studies comparing SSRIs to CBT reported inconclusive results for the proportion of participants who achieved complete remission of depressive symptoms. Seizure frequency data did not suggest an increased risk of seizures with antidepressants compared to control treatments or baseline. Two studies measured quality of life; antidepressants did not appear to improve quality of life over control. No studies reported on cognitive functioning. Two RCTs and one NRSI reported comparative data on adverse events; antidepressants did not appear to increase the severity or number of adverse events compared to controls. The NSRIs reported higher rates of withdrawals due to adverse events than lack of efficacy. Reported adverse events for antidepressants included nausea, dizziness, sedation, headache, gastrointestinal disturbance, insomnia, and sexual dysfunction. AUTHORS' CONCLUSIONS: Existing evidence on the effectiveness of antidepressants in treating depressive symptoms associated with epilepsy is still very limited. Rates of response to antidepressants were highly variable. There is low certainty evidence from one small RCT (64 participants) that venlafaxine may improve depressive symptoms more than no treatment; this evidence is limited to treatment between 8 and 16 weeks, and does not inform longer-term effects. Moderate to low evidence suggests neither an increase nor exacerbation of seizures with SSRIs. There are no available comparative data to inform the choice of antidepressant drug or classes of drug for efficacy or safety for treating people with epilepsy and depression. RCTs of antidepressants utilising interventions from other treatment classes besides SSRIs, in large samples of patients with epilepsy and depression, are needed to better inform treatment policy. Future studies should assess interventions across a longer treatment duration to account for delayed onset of action, sustainability of treatment responses, and to provide a better understanding of the impact on seizure control.


Subversion of integrins by enteropathogenic Yersinia.
Enteropathogenic Yersinia are gram-negative bacterial species that translocate from the lumen of the intestine and are able to grow within deep tissue sites. During the earliest stages of disease, the organism is able to bind integrin receptors that are presented on the apical surface of M cells in the intestine, which allows its internalization and subsequent translocation into regional lymph nodes. The primary integrin substrate is the outer-membrane protein invasin, which binds with extraordinarily high affinity to at least five different integrins that have the (beta)(1) chain. Bacterial uptake into host cells is modulated by the affinity of receptor-substrate interaction, receptor concentration and the ability of the substrate to aggregate target receptors.


Assessing Photoreceptor Structure in Retinitis Pigmentosa and Usher Syndrome.
The purpose of this study was to examine cone photoreceptor structure in retinitis pigmentosa (RP) and Usher syndrome using confocal and nonconfocal split-detector adaptive optics scanning light ophthalmoscopy (AOSLO). Nineteen subjects (11 RP, 8 Usher syndrome) underwent ophthalmic and genetic testing, spectral-domain optical coherence tomography (SD-OCT), and AOSLO imaging. Split-detector images obtained in 11 subjects (7 RP, 4 Usher syndrome) were used to assess remnant cone structure in areas of altered cone reflectivity on confocal AOSLO. Despite normal interdigitation zone and ellipsoid zone appearance on OCT, foveal and parafoveal cone densities derived from confocal AOSLO images were significantly lower in Usher syndrome compared with RP. This was due in large part to an increased prevalence of non-waveguiding cones in the Usher syndrome retina. Although significantly correlated to best-corrected visual acuity and foveal sensitivity, cone density can decrease by nearly 38% before visual acuity becomes abnormal. Aberrantly waveguiding cones were noted within the transition zone of all eyes and corresponded to intact inner segment structures. These remnant cones decreased in density and increased in diameter across the transition zone and disappeared with external limiting membrane collapse. Foveal cone density can be decreased in RP and Usher syndrome before visible changes on OCT or a decline in visual function. Thus, AOSLO imaging may allow more sensitive monitoring of disease than current methods. However, confocal AOSLO is limited by dependence on cone waveguiding, whereas split-detector AOSLO offers unambiguous and quantifiable visualization of remnant cone inner segment structure. Confocal and split-detector thus offer complementary insights into retinal pathology.


Increasing Uptake of HIV, Sexually Transmitted Infection, and Family Planning Services, and Reducing HIV-Related Risk Behaviors Among Youth Living With HIV in Uganda.
To address barriers to care for youth living with HIV (YLHIV), the Link Up project implemented a peer-led intervention model that provided a comprehensive package of HIV and sexual and reproductive health and rights services through community-based peer support groups for YLHIV. Peer educators delivered targeted counseling and health education, and referred YLHIV to antiretroviral therapy (ART), and reproductive health services that were available at youth-oriented sexual and reproductive health and rights facilities. At baseline (October to November 2014), 37 peer support groups for YLHIV were established in Luwero and Nakasongola districts. During this same time period, we recruited a cohort of 473 support group members, aged 15-24 years. After a 9-month intervention period (January to September 2015), we completed the end-line survey with 350 members of the original cohort. Multivariate logistic regression analysis applied to longitudinal data was used to assess changes in key outcomes from baseline to end line. Multivariate analyses showed significant increases at end line, compared with baseline, in self-efficacy (adjusted odds ratio [AOR]: 1.8 [1.3-2.6]), comprehensive HIV knowledge [AOR: 1.8 [1.3-2.6]), HIV disclosure (AOR: 1.6 [1.01-2.6]), condom use at last sex (AOR: 1.7 [1.2-2.5]), sexually transmitted infection uptake (AOR: 2.1 [1.5-2.9]), ART uptake (AOR: 2.5 [1.6-4.0]), ART adherence (AOR: 2.5 [1.3-4.9]), CD4 testing (AOR: 2.4 [1.5-3.6]), and current use of a modern contraceptive method (AOR: 1.7 [1.1-2.7]). Link Up's intervention strategy likely contributed to observed increases in self-efficacy, knowledge of HIV, condom use, HIV disclosure ART utilization and adherence, CD4 testing, STI testing uptake, and use of modern family planning methods. This model shows promise and should be adapted for use among YLHIV in similar settings and evaluated further.


Comparison of Choroidal Thickness Measurements Using Spectral Domain Optical Coherence Tomography in Six Different Settings and With Customized Automated Segmentation Software.
We investigate which spectral domain-optical coherence tomography (SD-OCT) setting is superior when measuring subfoveal choroidal thickness (CT) and compared results to an automated segmentation software. Thirty patients underwent enhanced depth imaging (EDI)-OCT. B-scans were extracted in six different settings (W+N = white background/normal contrast 9; W+H = white background/maximum contrast 16; B+N = black background/normal contrast 12; B+H = black background/maximum contrast 16; C+N = Color-encoded image on black background at predefined contrast of 9, and C+H = Color-encoded image on black background at high/maximal contrast of 16), resulting in 180 images. Subfoveal CT was manually measured by nine graders and by automated segmentation software. Intraclass correlation (ICC) was assessed. ICC was higher in normal than in high contrast images, and better for achromatic black than for white background images. Achromatic images were better than color images. Highest ICC was achieved in B+N (ICC = 0.64), followed by B+H (ICC = 0.54), W+N, and W+H (ICC = 0.5 each). Weakest ICC was obtained with Spectral-color (ICC = 0.47). Mean manual CT versus mean computer estimated CT showed a correlation of r = 0.6 (P = 0.001). Black background with white image at normal contrast (B+N) seems the best setting to manually assess subfoveal CT. Automated assessment of CT seems to be a reliable tool for CT assessment. To define optimized OCT analysis settings to improve the evaluation of in vivo imaging.


Protein profiling of angiogenesis-related growth factors in laryngeal carcinoma: Pattern of protein expression in relation to tumour progression.
The expression of angiogenesis-related proteins was determined in laryngeal tumour tissue, associated tumour involved lymph nodes, apparently normal mucosa and control tissue and were related to tumour stage. Both laryngeal tumour tissue and associated metastatic nodes were obtained from seven patients undergoing surgical resection; in four cases apparently normal mucosa was also dissected from the tumour specimen margins. Control uvula mucosa was obtained from five healthy volunteers undergoing uvulopalatopharyngoplasty. The relative expression of 55 angiogenesis-related proteins was determined in tissue lysates using a Proteome Profiler human angiogenesis array kit. The level of 32/55 angiogenesis-related proteins was higher in tumour tissue compared with controls. Furthermore, in these tumour biopsies higher levels of proteins were associated with increasing tumour stage. A similar trend was seen for 29/32 of these proteins in the nodal tissue. In T4 stage tumour tissue samples, 29/55 angiogenensis-related proteins were more highly expressed compared with the adjacent normal mucosa from the same patient, and this decreased to 8 proteins in tumour tissue from the T1 stage patients. In contrast, the expression of 23 angiogenesis-related proteins in metastatic lymph node tissue from T4 stage patients was lower compared with that found in the normal mucosa adjacent to the tumour. In conclusion, this study has identified a number of factors involved in angiogenesis that are likely to contribute to the growth and metastasis of laryngeal tumours. Furthermore, a number of factors were also substantially altered in metastatic deposits compared with the primary tumour mass or adjacent normal tissue. This study requires confirmatory analysis of the selected key factors in a larger cohort of patients.


AWE-WQ: fast-forwarding molecular dynamics using the accelerated weighted ensemble.
A limitation of traditional molecular dynamics (MD) is that reaction rates are difficult to compute. This is due to the rarity of observing transitions between metastable states since high energy barriers trap the system in these states. Recently the weighted ensemble (WE) family of methods have emerged which can flexibly and efficiently sample conformational space without being trapped and allow calculation of unbiased rates. However, while WE can sample correctly and efficiently, a scalable implementation applicable to interesting biomolecular systems is not available. We provide here a GPLv2 implementation called AWE-WQ of a WE algorithm using the master/worker distributed computing WorkQueue (WQ) framework. AWE-WQ is scalable to thousands of nodes and supports dynamic allocation of computer resources, heterogeneous resource usage (such as central processing units (CPU) and graphical processing units (GPUs) concurrently), seamless heterogeneous cluster usage (i.e., campus grids and cloud providers), and support for arbitrary MD codes such as GROMACS, while ensuring that all statistics are unbiased. We applied AWE-WQ to a 34 residue protein which simulated 1.5 ms over 8 months with peak aggregate performance of 1000 ns/h. Comparison was done with a 200 μs simulation collected on a GPU over a similar timespan. The folding and unfolded rates were of comparable accuracy.


Results of an innovative university-based recovery education program for adults with psychiatric disabilities.
This study examined the effectiveness of an educational approach to psychiatric rehabilitation called the Recovery Center. Using a quasi-experimental design we recruited 97 intervention and 81 comparison participants and examined the intervention's impact on health, mental health, subjective, and role functioning outcomes. Results suggested that this intervention was effective in improving subjective outcomes, especially empowerment and recovery attitudes, both of which received primary emphasis in the intervention. The Recovery Center, which integrates a bio-psychosocial framework with psycho-educational interventions shows promise as a complement to traditional mental health services in developing readiness for rehabilitation and promoting recovery among individuals with severe psychiatric disabilities.


De novo myocardial regeneration: advances and pitfalls.
The capability of adult tissue-derived stem cells for cardiogenesis has been extensively studied in experimental animals and clinical studies for treatment of postischemic cardiomyopathy. The less-than-anticipated improvement in the heart function in most clinical studies with skeletal myoblasts and bone marrow cells has warranted a search for alternative sources of stem cells. Despite their multilineage differentiation potential, ethical issues, teratogenicity, and tissue rejection are main obstacles in developing clinically feasible methods for embryonic stem cell transplantation into patients. A decade-long research on embryonic stem cells has paved the way for discovery of alternative approaches for generating pluripotent stem cells. Genetic manipulation of somatic cells for pluripotency genes reprograms the cells to pluripotent status. Efforts are currently focused to make reprogramming protocols safer for clinical applications of the reprogrammed cells. We summarize the advancements and complicating features of stem cell therapy and discuss the decade-and-a-half-long efforts made by stem cell researchers for moving the field from bench to the bedside as an adjunct therapy or as an alternative to the contemporary therapeutic modalities for routine clinical application. The review also provides a special focus on the advancements made in the field of somatic cell reprogramming.


Time Course Analysis of Skeletal Muscle Pathology of GDE5 Transgenic Mouse.
Glycerophosphodiesterase 5 (GDE5) selectively hydrolyses glycerophosphocholine to choline and is highly expressed in type II fiber-rich skeletal muscles. We have previously generated that a truncated mutant of GDE5 (GDE5dC471) that lacks phosphodiesterase activity and shown that transgenic mice overexpressing GDE5dC471 in skeletal muscles show less skeletal muscle mass than control mice. However, the molecular mechanism and pathophysiological features underlying decreased skeletal muscle mass in GDE5dC471 mice remain unclear. In this study, we characterized the skeletal muscle disorder throughout development and investigated the primary cause of muscle atrophy. While type I fiber-rich soleus muscle mass was not altered in GDE5dC471 mice, type II fiber-rich muscle mass was reduced in 8-week-old GDE5dC471 mice. Type II fiber-rich muscle mass continued to decrease irreversibly in 1-year-old transgenic mice with an increase in apoptotic cell. Adipose tissue weight and blood triglyceride levels in 8-week-old and 1-year-old transgenic mice were higher than those in control mice. This study also demonstrated compensatory mRNA expression of neuromuscular junction (NMJ) components, including nicotinic acetylcholine receptors (α1, γ, and ε subunits) and acetylcholinesterase in type II fiber-rich quadriceps muscles in GDE5dC471 mice. However, we did not observe morphological changes in NMJs associated with skeletal muscle atrophy in GDE5dC471 mice. We also found that HSP70 protein levels are significantly increased in the skeletal muscles of 2-week-old GDE5dC471 mice and in mouse myoblastic C2C12 cells overexpressing GDE5dC471. These findings suggest that GDE5dC471 mouse is a novel model of early-onset irreversible type II fiber-rich myopathy associated with cellular stress.


Sentinel node biopsy using a magnetic tracer versus standard technique: the SentiMAG Multicentre Trial.
The SentiMAG Multicentre Trial evaluated a new magnetic technique for sentinel lymph node biopsy (SLNB) against the standard (radioisotope and blue dye or radioisotope alone). The magnetic technique does not use radiation and provides both a color change (brown dye) and a handheld probe for node localization. The primary end point of this trial was defined as the proportion of sentinel nodes detected with each technique (identification rate). A total of 160 women with breast cancer scheduled for SLNB, who were clinically and radiologically node negative, were recruited from seven centers in the United Kingdom and The Netherlands. SLNB was undertaken after administration of both the magnetic and standard tracers (radioisotope with or without blue dye). A total of 170 SLNB procedures were undertaken on 161 patients, and 1 patient was excluded, leaving 160 patients for further analysis. The identification rate was 95.0 % (152 of 160) with the standard technique and 94.4 % (151 of 160) with the magnetic technique (0.6 % difference; 95 % upper confidence limit 4.4 %; 6.9 % discordance). Of the 22 % (35 of 160) of patients with lymph node involvement, 16 % (25 of 160) had at least 1 macrometastasis, and 6 % (10 of 160) had at least a micrometastasis. Another 2.5 % (4 of 160) had isolated tumor cells. Of 404 lymph nodes removed, 297 (74 %) were true sentinel nodes. The lymph node retrieval rate was 2.5 nodes per patient overall, 1.9 nodes per patient with the standard technique, and 2.0 nodes per patient with the magnetic technique. The magnetic technique is a feasible technique for SLNB, with an identification rate that is not inferior to the standard technique.


Immune-neural connections: how the immune system's response to infectious agents influences behavior.
Humans and animals use the classical five senses of sight, sound, touch, smell and taste to monitor their environment. The very survival of feral animals depends on these sensory perception systems, which is a central theme in scholarly research on comparative aspects of anatomy and physiology. But how do all of us sense and respond to an infection? We cannot see, hear, feel, smell or taste bacterial and viral pathogens, but humans and animals alike are fully aware of symptoms of sickness that are caused by these microbes. Pain, fatigue, altered sleep pattern, anorexia and fever are common symptoms in both sick animals and humans. Many of these physiological changes represent adaptive responses that are considered to promote animal survival, and this constellation of events results in sickness behavior. Infectious agents display a variety of pathogen-associated molecular patterns (PAMPs) that are recognized by pattern recognition receptors (PRRs). These PRR are expressed on both the surface [e.g. Toll-like receptor (TLR)-4] and in the cytoplasm [e.g. nucleotide-binding oligomerization domain (Nod)-like receptors] of cells of the innate immune system, primarily macrophages and dendritic cells. These cells initiate and propagate an inflammatory response by stimulating the synthesis and release of a variety of cytokines. Once an infection has occurred in the periphery, both cytokines and bacterial toxins deliver this information to the brain using both humoral and neuronal routes of communication. For example, binding of PRR can lead to activation of the afferent vagus nerve, which communicates neuronal signals via the lower brain stem (nucleus tractus solitarius) to higher brain centers such as the hypothalamus and amygdala. Blood-borne cytokines initiate a cytokine response from vascular endothelial cells that form the blood-brain barrier (BBB). Cytokines can also reach the brain directly by leakage through the BBB via circumventricular organs or by being synthesized within the brain, thus forming a mirror image of the cytokine milieu in the periphery. Although all cells within the brain are capable of initiating cytokine secretion, microglia have an early response to incoming neuronal and humoral stimuli. Inhibition of proinflammatory cytokines that are induced following bacterial infection blocks the appearance of sickness behaviors. Collectively, these data are consistent with the notion that the immune system communicates with the brain to regulate behavior in a way that is consistent with animal survival.


Intratumoral heterogeneity in a minority of ovarian low-grade serous carcinomas.
Ovarian low-grade serous carcinoma (LGSC) has fewer mutations than ovarian high-grade serous carcinoma (HGSC) and a less aggressive clinical course. However, an overwhelming majority of LGSC patients do not respond to conventional chemotherapy resulting in a poor long-term prognosis comparable to women diagnosed with HGSC. KRAS and BRAF mutations are common in LGSC, leading to clinical trials targeting the MAPK pathway. We assessed the stability of targetable somatic mutations over space and/or time in LGSC, with a view to inform stratified treatment strategies and clinical trial design. Eleven LGSC cases with primary and recurrent paired samples were identified (stage IIB-IV). Tumor DNA was isolated from 1-4 formalin-fixed paraffin-embedded tumor blocks from both the primary and recurrence (n = 37 tumor and n = 7 normal samples). Mutational analysis was performed using the Ion Torrent AmpliSeqTM Cancer Panel, with targeted validation using Fluidigm-MiSeq, Sanger sequencing and/or Raindance Raindrop digital PCR. KRAS (3/11), BRAF (2/11) and/or NRAS (1/11) mutations were identified in five unique cases. A novel, non-synonymous mutation in SMAD4 was observed in one case. No somatic mutations were detected in the remaining six cases. In two cases with a single matched primary and recurrent sample, two KRAS hotspot mutations (G12V, G12R) were both stable over time. In three cases with multiple samplings from both the primary and recurrent surgery some mutations (NRAS Q61R, BRAF V600E, SMAD4 R361G) were stable across all samples, while others (KRAS G12V, BRAF G469V) were unstable. Overall, the majority of cases with detectable somatic mutations showed mutational stability over space and time while one of five cases showed both temporal and spatial mutational instability in presumed drivers of disease. Investigation of additional cases is required to confirm whether mutational heterogeneity in a minority of LGSC is a general phenomenon that should be factored into the design of clinical trials and stratified treatment for this patient population.


Economic linkages to changing landscapes.
Many economic processes are intertwined with landscape change. A large number of individual economic decisions shape the landscape, and in turn the changes in the landscape shape economic decisions. This article describes key research questions about the economics of landscape change and reviews the state of research knowledge. The rich and varied economic-landscape interactions are an active area of research by economists, geographers, and others. Because the interactions are numerous and complex, disentangling the causal relationships in any given landscape system is a formidable research challenge. Limited data with mismatched temporal and spatial scales present further obstacles. Nevertheless, the growing body of economic research on these topics is advancing and shares fundamental challenges, as well as data and methods, with work in other disciplines.


Management of parapneumonic effusions and empyema.
Pleural infection remains a common and serious respiratory condition with important implications for patients and health-care services. This review will cover the management of pleural infection including medical treatment, the role of intrapleural agents and surgical treatment. We discuss the directions that future research in this important area might take. Increasing incidence of pleural infection has been reported worldwide without a clear explanation. The pathogens responsible for pleural infection differ from those in pneumonia. Proper antibiotic selection and pleural fluid drainage remain the cornerstones of treatment. There is no evidence in adult pleural infection to support the routine use of intrapleural fibrinolytics to alter clinically meaningful outcomes; however, combined intrapleural tissue plasminogen activator (tPA) and deoxyribonuclease (DNase) therapy may have a future role. The role of medical thoracoscopy remains unproven. Surgical referral should be considered in patients who fail to respond to standard medical management after 5 to 7 days. Despite advances in microbiology, medical management, and surgery, the mortality of pleural infection at one year remains approximately 20% for the last two decades. Future studies are required to validate predictive scores for patients' stratification (RAPID score) and the role of fibrinolytics (combination of tPA plus DNase). Surgical drainage remains a vital treatment option, but ongoing research is required to define the group of patients who would benefit most and when, in the disease course, this treatment should be offered.


Tethering of Multi-Vesicular Bodies and the Tonoplast to the Plasma Membrane in Plants.
Tethering of the plasma membrane (PM) and many organelles to the endoplasmic reticulum (ER) for communication and lipid exchange has been widely reported. However, despite growing interest in multi-vesicular bodies (MVBs) as possible sources of exosomes, tethering of MVBs to the PM has not been reported. Here we show that MVBs and the vacuolar membrane (tonoplast) could be tethered to the PM (PM-MVB/TP tethering) by artificial protein fusions or bimolecular fluorescence complementation (BiFC) complexes that contain a peripheral membrane protein that binds the PM and also a protein that binds MVBs or the tonoplast. PM-binding proteins capable of participating in PM-MVB/TP tethering included StRem1.3, BIK1, PBS1, CPK21, and the PtdIns(4)-binding proteins FAPP1 and Osh2. MVB/TP-binding proteins capable of participating in tethering included ARA6, ARA7, RHA1, RABG3f, and the PtdIns(3)P-binding proteins Vam7p and Hrs-2xFYVE. BiFC complexes or protein fusions capable of producing PM-MVB/TP tethering were visualized as large well-defined patches of fluorescence on the PM that could displace PM proteins such as AtFlotillin1 and also could displace cytoplasmic proteins such as soluble GFP. Furthermore, we identified paralogous ubiquitin E3 ligase proteins, SAUL1 (AtPUB44), and AtPUB43 that could produce PM-MVB/TP tethering. SAUL1 and AtPUB43 could produce tethering in uninfected tissue when paired with MVB-binding proteins or when their E3 ligase domain was deleted. When Nicotiana benthamiana leaf tissue was infected with Phytophthora capsici, full length SAUL1 and AtPUB43 localized in membrane patches consistent with PM-MVB/TP tethering. Our findings define new tools for studying PM-MVB/TP tethering and its possible role in plant defense. Although not previously observed, the tethering of multi-vesicular bodies to the plasma membrane is of interest due to the potential role of this process in producing exosomes in plants. Here we describe tools for observing and manipulating the tethering of multi-vesicular bodies and the tonoplast to the plant plasma membrane, and describe two plant proteins that may naturally regulate this process during infection.


Systematic Comparison of Retinal Organoid Differentiation from Human Pluripotent Stem Cells Reveals Stage Specific, Cell Line, and Methodological Differences.
A major goal in the stem cell field is to generate tissues that can be utilized as a universal tool for in vitro models of development and disease, drug development, or as a resource for patients suffering from disease or injury. Great efforts are being made to differentiate human pluripotent stem cells in vitro toward retinal tissue, which is akin to native human retina in its cytoarchitecture and function, yet the numerous existing retinal induction protocols remain variable in their efficiency and do not routinely produce morphologically or functionally mature photoreceptors. Herein, we determine the impact that the method of embryoid body (EB) formation and maintenance as well as cell line background has on retinal organoid differentiation from human embryonic stem cells and human induced pluripotent stem cells. Our data indicate that cell line-specific differences dominate the variables that underline the differentiation efficiency in the early stages of differentiation. In contrast, the EB generation method and maintenance conditions determine the later differentiation and maturation of retinal organoids. Of the latter, the mechanical method of EB generation under static conditions, accompanied by media supplementation with Y27632 for the first 48 hours of differentiation, results in the most consistent formation of laminated retinal neuroepithelium containing mature and electrophysiologically responsive photoreceptors. Collectively, our data provide substantive evidence for stage-specific differences in the ability to give rise to laminated retinae, which is determined by cell line-specific differences in the early stages of differentiation and EB generation/organoid maintenance methods at later stages.


Exogenous collagen cross-linking recovers tendon functional integrity in an experimental model of partial tear.
We investigated the hypothesis that exogenous collagen cross-linking can augment intact regions of tendon to mitigate mechanical propagation of partial tears. We first screened the low toxicity collagen cross-linkers genipin, methylglyoxal and ultra-violet (UV) light for their ability to augment tendon stiffness and failure load in rat tail tendon fascicles (RTTF). We then investigated cross-linking effects in load bearing equine superficial digital flexor tendons (SDFT). Data indicated that all three cross-linking agents augmented RTTF mechanical properties but reduced native viscoelasticity. In contrast to effects observed in fascicles, methylglyoxal treatment of SDFT detrimentally affected tendon mechanical integrity, and in the case of UV did not alter tendon mechanics. As in the RTTF experiments, genipin cross-linking of SDFT resulted in increased stiffness, higher failure loads and reduced viscoelasticity. Based on this result we assessed the efficacy of genipin in arresting tendon tear propagation in cyclic loading to failure. Genipin cross-linking secondary to a mid-substance biopsy-punch significantly reduced tissue strains, increased elastic modulus and increased resistance to fatigue failure. We conclude that genipin cross-linking of injured tendons holds potential for arresting tendon tear progression, and that implications of the treatment on matrix remodeling in living tendons should now be investigated.


Micronutrient status and human immunodeficiency virus (HIV) infection.
This study surveyed serum concentrations of vitamins, electrolytes, and trace elements in subjects seropositive for HIV-1 by ELISA and confirmatory Western blot. Thirty subjects (26 males, 4 females) were recruited at a hospital clinic. Seventeen were classified as having mild or severe ARC (AIDS-related complex), 7 had AIDS, and 6 were asymptomatic. Eight had experienced weight loss of 10 pounds or more in the past 6 months. Most (93%) were anergic to skin test antigens. Percentages of subjects with below normal plasma concentrations include: zinc-30%, calcium-27%, magnesium-30%, carotenes-31%, total choline-50%, and ascorbate-27%. Eighty-seven percent of the subjects had at least one abnormally low value. Percentages with above normal values include: folate-37% and carnitine-37%. Some subjects with above normal values for plasma vitamins reported self-supplementation, usually with large doses. The results suggest that one or more abnormally low concentrations of the plasma micronutrients studied here are likely to be present in the majority of HIV seropositive patients.


Endocrine disrupting chemicals research program of the U.S. Environmental Protection Agency: summary of a peer-review report.
At the request of the U.S. Environmental Protection Agency (EPA) Office of Research and Development, a subcommittee of the Board of Scientific Counselors Executive Committee conducted an independent and open peer review of the Endocrine Disrupting Chemicals Research Program (EDC Research Program) of the U.S. EPA. The subcommittee was charged with reviewing the design, relevance, progress, scientific leadership, and resources of the program. The subcommittee found that the long-term goals and science questions in the EDC Program are appropriate and represent an understandable and solid framework for setting research priorities, representing a combination of problem-driven and core research. Long-term goal (LTG) 1, dealing with the underlying science surrounding endocrine disruptors, provides a solid scientific foundation for conducting risk assessments and making risk management decisions. LTG 2, dealing with defining the extent of the impact of endocrine-disrupting chemicals (EDCs), has shown greater progress on ecologic effects of EDCs compared with that on human health effects. LTG 3, which involves support of the Endocrine Disruptor Screening and Testing Program of the U.S. EPA, has two mammalian tests already through a validation program and soon available for use. Despite good progress, we recommend that the U.S. EPA a) strengthen their expertise in wildlife toxicology, b) expedite validation of the Endocrine Disruptors Screening and Testing Advisory Committee tests, c) continue dependable funding for the EDC Research Program, d) take a leadership role in the application of "omics" technologies to address many of the science questions critical for evaluating environmental and human health effects of EDCs, and e) continue to sponsor multidisciplinary intramural research and interagency collaborations.


Severe Intracranial Involvement in Giant Cell Arteritis: 5 Cases and Literature Review.
Involvement of intracranial arteries in giant cell arteritis (GCA) is rare. We describe the neurologic complications of intracranial GCA (IC GCA) and available treatment options. We describe 5 IC GCA cases from 3 Canadian vasculitis centers and review the literature. We searched English-language publications reporting similar patients meeting American College of Rheumatology (ACR) criteria for GCA and having intracranial artery involvement diagnosed by autopsy, magnetic resonance angiography, computed tomography angiography, or conventional angiography. All 5 cases of IC GCA met ACR criteria for GCA; 4 cases had a temporal artery biopsy that was consistent with GCA. All cases experienced cerebrovascular accident(s). Arteritis involved the following vessels: intracranial internal carotid (n = 1), vertebrobasilar arteries (n = 1), or both (n = 3). All cases received aspirin and oral prednisone (preceded by intravenous methylprednisone in 3 cases), combined with an immunosuppressant in 4 cases. All patients survived; 2 had complete neurological recovery, 3 had residual neurologic sequelae. The literature review included 42 cases from 28 publications. The clinical features of the reported cases were similar to those of our 5 cases. However, mortality was 100% in untreated cases (n = 2), 58% in those treated with corticosteroid alone (n = 31), and 40% in those treated with corticosteroid and an immunosuppressant (n = 10). IC GCA appears to be associated with neurologic complications and mortality. In some cases corticosteroid alone was not sufficient to prevent neurologic complications. The role of additional immunosuppressive agents needs further investigation.


Endothelial cell decay after descemet's stripping automated endothelial keratoplasty and top hat penetrating keratoplasty.
To analyze endothelial cell density (ECD) decay after Descemet's stripping automated endothelial keratoplasty (DSAEK) and top hat keratoplasty (THPK) in patients with Fuchs' endothelial dystrophy (FED) and/or pseudophakic bullous keratopathy (PPBK). Patients underwent either THPK (n = 33) or DSAEK (n = 39) at the Erasmus Medical Center, Rotterdam. For each nonrandomized cohort, a biexponential regression model for ECD decay was fitted. Factors associated with higher ECD decay were evaluated. Median follow-up was 31.2 months (range, 11-91) in the THPK cohort, and 23.4 months (range, 6-61) in the DSAEK group. The early ECD decay was much higher after DSAEK (half time, 2.2 months) than after THPK (half time, 12.8 months). The late ECD decay after DSAEK was less steep (half time, 75.5 months) than after THPK (half time, 62 months). The 1-, 3- and 5-year endothelial cell losses derived from the models after DSAEK were 56%, 66%, and 73%, respectively, and after THPK were 24%, 50%, and 64%, respectively. For the DSAEK cohort, PPBK as an indication for surgery was associated with significantly higher late-phase decay rates. For the top-hat cohort, a significantly lower late-phase decay rate was found in PPBK. FED and same-session cataract surgery were confounding variables in the DSAEK cohort. Regarding DSAEK, postoperative re-bubbling was not found to have significant effects on early or late ECD decay rates. However, the small sample size and other limitations related to the method of evaluation may have influenced these findings. After DSAEK, early ECD decay was stronger than after THPK, as opposed to late decay. Late decay was faster for PPBK than for FED after DSAEK.


Tactile numerosity is coded in external space.
Humans, and several non-human species, possess the ability to make approximate but reliable estimates of the number of objects around them. Alike other perceptual features, numerosity perception is susceptible to adaptation: exposure to a high number of items causes underestimation of the numerosity of a subsequent set of items, and vice versa. Several studies have investigated adaptation in the auditory and visual modality, whereby stimuli are preferentially encoded in an external coordinate system. As tactile stimuli are primarily coded in an internal (body-centered) reference frame, here we ask whether tactile numerosity adaptation operates based on internal or external spatial coordinates as it occurs in vision or audition. Twenty participants performed an adaptation task with their right hand located either in the right (uncrossed) or left (crossed) hemispace, in order for the two hands to occupy either two completely different positions, or the same position in space, respectively. Tactile adaptor and test stimuli were passively delivered either to the same (adapted) or different (non-adapted) hands. Our results show a clear signature of tactile numerosity adaptation aftereffects with a pattern of over- and under-estimation according to the adaptation rate (low and high, respectively). In the uncrossed position, we observed stronger adaptation effects when adaptor and test stimuli were delivered to the "adapted" hand. However, when both hands were aligned in the same spatial position (crossed condition), the magnitude of adaptation was similar irrespective of which hand received adaptor and test stimuli. These results demonstrate that numerosity information is automatically coded in external coordinates even in the tactile modality, suggesting that such a spatial reference frame is an intrinsic property of numerosity processing irrespective of the sensory modality.


Association of pulmonary artery obstruction index with elevated heart-type fatty acid binding protein and short-term mortality in patients with pulmonary embolism at intermediate risk.
Heart-type fatty acid binding protein (H-FABP) is a sensitive marker of myocardial injury and predictor of worse prognosis in patients with pulmonary embolism (PE). Assessment of right ventricular dysfunction and pulmonary artery obstruction index (PAOI) with computed tomography (CT) has been reported as a predictor of mortality in PE. Therefore, we aimed to assess the correlation between H-FABP and CT angiographic PAOI in PE patients at intermediate risk. Sixty-one patients (28 males; mean age, 62 ± 17 years) with diagnosis of PE were included in this study. CT was performed in all patients, and the following parameters were evaluated: right ventricle/left ventricle ratio (RV/LV), pulmonary artery axial diameter, superior vena cava axial diameter, and PAOI determined with Qanadli score. Blood samples were assessed for H-FABP and troponin levels. Patients were followed for 30 days after discharge. Mean PAOI was 57 ± 18%. Eleven patients died during the follow-up period due to PE (18% mortality rate). H-FABP was positive in 21 patients (35%). There was no difference in CT parameters between patients with positive H-FABP and negative H-FABP. In addition, CT parameters were similar between patients who survived and those who did not. RV/LV ratio correlated with PAOI score. Among the biomarkers, troponin levels correlated with both RV/LV ratio and PAOI. H-FABP was an independent predictor of mortality. PAOI and RV/LV ratio did not predict 30-day mortality. Although H-FABP positivity confers a bad prognosis on PE patients at intermediate risk, PAOI did not predict mortality in this group.


The Prevalence of Scabies and Impetigo in the Solomon Islands: A Population-Based Survey.
Scabies and impetigo are common, important and treatable skin conditions. Reports from several Pacific island countries show extremely high prevalence of these two conditions, but for many countries, including the Solomon Islands, there is a paucity of epidemiological data. Ten rural villages in the Western Province of the Solomon Islands were included in the study, chosen so that data collection could be integrated with an existing project investigating clinical and serological markers of yaws. All residents were eligible to participate, and 1908 people were enrolled. Participants were interviewed and examined by a paediatric registrar, who recorded relevant demographic information, and made a clinical diagnosis of scabies and/or impetigo, severity and distribution. The total unweighted prevalence of scabies was 19.2% (95% confidence interval [CI] 17.5-21.0), and age and gender weighted prevalence 19.2% (95%CI 16.7-21.9). The adult prevalence of scabies was 10.4% (95%CI 8.2-13.2), and the highest prevalence was found in infants < 1 year of age (34.1%, adjusted odds ratio [AOR] compared with adults: 3.6, 95%CI 2.2-6.0) and children aged 1-4 years (25.7%, AOR 2.6, 95%CI 1.7-3.9). Scabies affected two or more body regions in 80.9% of participants, and 4.4% of scabies cases were classified as severe. The total unweighted prevalence of active impetigo was 32.7% (95%CI 30.6-34.8), and age and gender weighted prevalence 26.7% (95%CI 24.2-29.5). The highest prevalence was found in children aged 1-4 years (42.6%, AOR compared with adults: 4.1, 95%CI 2.9-5.8). Scabies infestation was associated with active impetigo infection (AOR 2.0, 95%CI 1.6-2.6); with 41.1% of active impetigo cases also having scabies. Scabies and impetigo are very common in the rural Western Province of the Solomon Islands. Scabies infestation is strongly associated with impetigo. Community control strategies for scabies may reduce the burden of both conditions and their downstream complications.


Thermal landscape change as a driver of ectotherm responses to plant invasions.
A growing body of research demonstrates the impacts of invasive alien plants on native animals, but few studies consider thermal effects as a driver of the responses of native organisms. As invasive alien plants establish and alter the composition and arrangement of plant communities, the thermal landscapes available to ectotherms also change. Our study reviews the research undertaken to date on the thermal effects of alien plant invasions on native reptiles, amphibians, insects and arachnids. The 37 studies published between 1970 and early 2019 portray an overall detrimental effect of invasive plants on thermal landscapes, ectothermic individuals' performance and species abundance, diversity and composition. With a case study of a lizard species, we illustrate the use of thermal ecology tools in plant invasion research and test the generality of alien plant effects: changes in thermoregulation behaviour in invaded landscapes varied depending on the level of invasion and lizard traits. Together, the literature review and case study show that thermal effects of alien plants on ectotherms can be substantial albeit context-dependent. Further research should cover multiple combinations of native/invasive plant growth forms, invasion stages and ectotherm traits. More attention is also needed to test causality along the chain of effects from thermal landscapes to individuals, populations and communities.


Critical Research Needed to Examine the Environmental Impacts of Expanded Refrigeration on the Food System.
The unbroken global refrigerated supply chain, or cold chain, is rapidly expanding in developing countries. In addition to increasing the energy intensity of the food system, the expanded cold chain may facilitate changes in the global diet, food waste patterns, food production and distribution, and shopping habits. The sustainability impacts of many of these changes chain are unknown, given the complexity of interacting social, economic, and technical factors. The current literature surrounding the environmental impacts of refrigeration in the food system focuses on the direct impacts of energy use and coolant emissions, and lacks a critical evaluation of the accompanying systemic societal changes that potentially carry greater environmental impacts. This review examines the cold chain as a transformative technology, identifying key intrinsic, indirect, and external factors that will favorably, unfavorably, or ambiguously impact the environmental profile of the food system. The review identifies key interactions and feedbacks between the cold chain, food production and consumption decisions, infrastructure development, and the global environment which are largely unexamined and in need of empirical data. Viewing cold chain expansion from this broader perspective is essential to understanding the changing impacts of the food system in developing countries and may inform future sustainability planning.


Protein-coding variants implicate novel genes related to lipid homeostasis contributing to body-fat distribution.
Body-fat distribution is a risk factor for adverse cardiovascular health consequences. We analyzed the association of body-fat distribution, assessed by waist-to-hip ratio adjusted for body mass index, with 228,985 predicted coding and splice site variants available on exome arrays in up to 344,369 individuals from five major ancestries (discovery) and 132,177 European-ancestry individuals (validation). We identified 15 common (minor allele frequency, MAF ≥5%) and nine low-frequency or rare (MAF <5%) coding novel variants. Pathway/gene set enrichment analyses identified lipid particle, adiponectin, abnormal white adipose tissue physiology and bone development and morphology as important contributors to fat distribution, while cross-trait associations highlight cardiometabolic traits. In functional follow-up analyses, specifically in Drosophila RNAi-knockdowns, we observed a significant increase in the total body triglyceride levels for two genes (DNAH10 and PLXND1). We implicate novel genes in fat distribution, stressing the importance of interrogating low-frequency and protein-coding variants.


Mutation in NRAS in familial Noonan syndrome--case report and review of the literature.
Noonan syndrome (NS), a heterogeneous developmental disorder associated with variable clinical expression including short stature, congenital heart defect, unusual pectus deformity and typical facial features, is caused by activating mutations in genes involved in the RAS-MAPK signaling pathway. Here, we present a clinical and molecular characterization of a small family with Noonan syndrome. Comprehensive mutation analysis of NF1, PTPN11, SOS1, CBL, BRAF, RAF1, SHOC2, MAP2K2, MAP2K1, SPRED1, NRAS, HRAS and KRAS was performed using targeted next-generation sequencing. The result revealed a recurrent mutation in NRAS, c.179G > A (p.G60E), in the index patient. This mutation was inherited from the index patient's father, who also showed signs of NS. We describe clinical features in this family and review the literature for genotype-phenotype correlations for NS patients with mutations in NRAS. Neither of affected individuals in this family presented with juvenile myelomonocytic leukemia (JMML), which together with previously published results suggest that the risk for NS individuals with a germline NRAS mutation developing JMML is not different from the proportion seen in other NS cases. Interestingly, 50% of NS individuals with an NRAS mutation (including our family) present with lentigines and/or Café-au-lait spots. This demonstrates a predisposition to hyperpigmented lesions in NRAS-positive NS individuals. In addition, the affected father in our family presented with a hearing deficit since birth, which together with lentigines are two characteristics of NS with multiple lentigines (previously LEOPARD syndrome), supporting the difficulties in diagnosing individuals with RASopathies correctly. The clinical and genetic heterogeneity observed in RASopathies is a challenge for genetic testing. However, next-generation sequencing technology, which allows screening of a large number of genes simultaneously, will facilitate an early and accurate diagnosis of patients with RASopathies.


Whole-Body Vibration as a Modality for the Rehabilitation of Peripheral Neuropathies: Implications for Cancer Survivors Suffering from Chemotherapy-Induced Peripheral Neuropathy.
The objective was to study the effect of whole-body vibration (WBV) on strength, balance and pain in patients with peripheral neuropathies and to consider its significance for the rehabilitation of patients suffering from chemotherapy-induced peripheral neuropathy (CIPN). Using a broad search strategy, PubMed was searched for clinical trials on WBV interventions aimed at improving strength, balance or pain in patients with peripheral neuropathies, which were published in English until 5(th) June 2014. The search was performed by the first author and generated a total of 505 results, which yielded 5 articles that met the inclusion criteria, being studies: i) published in English; ii) involving adult human subjects' peripheral neuropathies; iii) evaluating the effect of WBV as a therapeutic intervention; and iv) reporting findings for at least one of the following outcomes: strength, balance or pain. Methodological quality of included studies was assessed independently by first and second author, using the physiotherapy evidence database scale. The overall methodological quality of included studies was low. Two studies found a beneficial effect of WBV on neuropathic pain, but another study failed to find the same effect. One study found significant improvements in both muscle strength and balance, while another study found improvements only in some, but not all, of the applied tests to measure muscle strength and balance. The results of this literature search suggest insufficient evidence to assess the effectiveness for the effects of WBV on neuropathic pain, muscle strength and balance in patients with peripheral neuropathies. More high-quality trials are needed to guide the optimization of rehabilitation programs for cancer survivors with CIPN in particular.


IL-17-high asthma with features of a psoriasis immunophenotype.
The role of IL-17 immunity is well established in patients with inflammatory diseases, such as psoriasis and inflammatory bowel disease, but not in asthmatic patients, in whom further study is required. We sought to undertake a deep phenotyping study of asthmatic patients with upregulated IL-17 immunity. Whole-genome transcriptomic analysis was performed by using epithelial brushings, bronchial biopsy specimens (91 asthmatic patients and 46 healthy control subjects), and whole blood samples (n = 498) from the Unbiased Biomarkers for the Prediction of Respiratory Disease Outcomes (U-BIOPRED) cohort. Gene signatures induced in vitro by IL-17 and IL-13 in bronchial epithelial cells were used to identify patients with IL-17-high and IL-13-high asthma phenotypes. Twenty-two of 91 patients were identified with IL-17, and 9 patients were identified with IL-13 gene signatures. The patients with IL-17-high asthma were characterized by risk of frequent exacerbations, airway (sputum and mucosal) neutrophilia, decreased lung microbiota diversity, and urinary biomarker evidence of activation of the thromboxane B2 pathway. In pathway analysis the differentially expressed genes in patients with IL-17-high asthma were shared with those reported as altered in psoriasis lesions and included genes regulating epithelial barrier function and defense mechanisms, such as IL1B, IL6, IL8, and β-defensin. The IL-17-high asthma phenotype, characterized by bronchial epithelial dysfunction and upregulated antimicrobial and inflammatory response, resembles the immunophenotype of psoriasis, including activation of the thromboxane B2 pathway, which should be considered a biomarker for this phenotype in further studies, including clinical trials targeting IL-17.


Classical Hodgkin Lymphoma with Reduced β2M/MHC Class I Expression Is Associated with Inferior Outcome Independent of 9p24.1 Status.
In classical Hodgkin lymphoma (cHL), malignant Hodgkin Reed-Sternberg (HRS) cells evade antitumor immunity by multiple mechanisms, including perturbed antigen presentation and enhanced PD-1 signaling. HRS cell expression of the PD-1 ligands is attributable, in part, to copy number alterations of 9p24.1/CD274(PD-L1)/PDCD1LG2(PD-L2) Amplification of PD-L1/PD-L2 is associated with advanced clinical stage and inferior progression-free survival (PFS) following first-line (induction) therapy. The relationships between altered expression of β2-microglobulin (β2M), MHC class I, and MHC class II by HRS cells, PD-L1/PD-L2 amplification, and clinical outcome in cHL are poorly defined. We assessed these variables in diagnostic biopsy specimens from 108 patients with cHL who received uniform treatment and had long-term follow-up and found decreased/absent expression of β2M/MHC class I in 79% (85/108) and decreased/absent expression of MHC class II in 67% (72/108) of cases. Patients with decreased/absent β2M/MHC class I had shorter PFS, independent of PD-L1/PD-L2 amplification and advanced stage. Decreased or absent MHC class II was unrelated to outcome. These results suggest that MHC class I-mediated antigen presentation by HRS cells is an important component of the biological response to standard chemo/radiotherapy. The paucity of β2M/MHC class I expression on HRS cells also prompts speculation regarding alternative mechanisms of action of PD-1 blockade in cHL. Cancer Immunol Res; 4(11); 910-6. ©2016 AACR.


An observational study of pain self-management strategies and outcomes: does type of pain, age, or gender, matter?
Background and aims Acute pain is differentiated from chronic pain by its sudden onset and short duration; in contrast, chronic pain is characterized by a duration of at least several months, typically considered longer than normal healing time. Despite differences in definition, there is little information on how types of self-management strategies or outcomes differ when pain is chronic rather than acute. Additionally, age and gender are thought to be related to types of strategies used and outcomes. However, strategies used and outcomes can be influenced by level of education, socioeconomic status, occupation, and access to the health care system, which can confound associations to type of pain, age or gender. The purpose of this study was to examine the association of strategies used for pain self-management and outcomes with type of pain, acute or chronic, age, or gender in a socioeconomically homogenous population, pharmacists. Methods Pharmacists with acute or chronic pain and a valid email completed an on-line questionnaire on demographic characteristics, pain characteristics, pharmacological and non-pharmacological strategies for managing pain, and outcomes (e.g. pain intensity). Univariate analysis was conducted by stratifying on type of pain (acute or chronic), then stratifying on gender (men vs. women) and age (younger vs. older). The a priori alpha level was 0.05. Results A total of 366 pharmacists completed the questionnaire, 212 with acute pain (average age=44±12.1; 36% men) and 154 with chronic pain (average age=53±14.0; 48% men). The chronic pain group reported substantially higher levels of pain before treatment, level of post-treatment pain, level of pain at which sleep was possible, and goal pain levels (effect sizes [ES's]=0.37-0.61). The chronic pain group were substantially more likely to use prescription non-steroidal anti-inflammatory medications (NSAIDS), opioids, and non-prescription pain relievers (ES's=0.29-0.80), and non-medical strategies (ES's=0.56-0.77). Participants with chronic pain also were less confident (ES=0.54) and less satisfied (ES=0.52). In contrast, there were no differences within either the acute or chronic pain groups related to gender and outcomes. In the acute pain group, there also were no gender differences related to management strategies. However, younger age in the acute pain group was associated with use of herbal remedies and use of rest. Within the chronic pain group, men were more likely to use NSAIDS and women more likely to use hot/cold packs or massage while older participants were more likely to use massage. Variability in post-treatment level of pain and percent relief was high in all groups (coefficient of variation=25%-100%). Conclusions The differences between acute and chronic pain were substantial and included differences in demographic characteristics, pain characteristics, management strategies used, and outcomes. In contrast, few associations between age and gender with either management strategies or outcomes were identified, although the variability was high. Implications When managing or researching pain management, acute pain should be differentiated from chronic pain. Because of the substantial variability within the gender and age groups, an individual approach to pain management irrespective of age and gender may be most useful.


LFA-1-specific therapy prolongs allograft survival in rhesus macaques.
Outcomes in transplantation have been limited by suboptimal long-term graft survival and toxicities associated with current immunosuppressive approaches. T cell costimulation blockade has shown promise as an alternative strategy to avoid the side effects of conventional immunosuppressive therapies, but targeting CD28-mediated costimulation alone has proven insufficient to prevent graft rejection in primates. Donor-specific memory T (TM) cells have been implicated in costimulation blockade-resistant transplant rejection, due to their enhanced effector function and decreased reliance on costimulatory signaling. Thus, we have tested a potential strategy to overcome TM cell-driven rejection by targeting molecules preferentially expressed on these cells, such as the adhesion molecule lymphocyte function-associated antigen 1 (LFA-1). Here, we show that short-term treatment (i.e., induction therapy) with the LFA-1-specific antibody TS-1/22 in combination with either basiliximab (an IL-2Rα-specific mAb) and sirolimus (a mammalian target of rapamycin inhibitor) or belatacept (a high-affinity variant of the CD28 costimulation-blocker CTLA4Ig) prolonged islet allograft survival in nonhuman primates relative to control treatments. Moreover, TS-1/22 masked LFA-1 on TM cells in vivo and inhibited the generation of alloproliferative and cytokine-producing effector T cells that expressed high levels of LFA-1 in vitro. These results support the use of LFA-1-specific induction therapy to neutralize costimulation blockade-resistant populations of T cells and further evaluation of LFA-1-specific therapeutics for use in transplantation.


A correlation of immunohistochemical expression of TP53 and severity of inflammation with varying grades of oral squamous cell carcinoma.
Epithelial cells exposed to carcinogens and genetic damage, once surpass reversible cell damage, either undergo apoptosis or transform into malignancy, chiefly oral squamous cell carcinoma (OSCC). Progressive accumulation of genetic errors in TP53 results in tumorigenesis. Inflammation is also a modulator in this process. The present study attempted to correlate the immunohistochemical expression of TP53 with increased aggressiveness of OSCC, to determine how these immune cells regulate the path of carcinogenesis and to define the role of inflammation in TP53 immunoexpression. Tissue sections from 24 biopsy-proven cases of OSCC were stained with anti-TP53 antibody. Five hundred neoplastic epithelial cells and inflammatory cells, each, were counted at invasive tumor front. Two hundred peritumoral neutrophils were counted based on nuclear lobes present. The least TP53 expression was seen in well-differentiated OSCC (P < 0.001), whereas neutrophil count and plasma cell count were found to be least in well-differentiated OSCC (P < 0.001), whereas the number of lymphocytes and macrophages decreased with increased grading of OSCC (P < 0.001). Four- and five-lobed neutrophils were found to be highest in poorly differentiated OSCC (P < 0.001), whereas two-lobed neutrophil count was seen to be maximum in well-differentiated OSCC (P < 0.001). TP53 plays a significant role in the initiation and progression of OSCC. Inflammation plays a role of friend and a foe simultaneously, and little modification in the present treatment modalities for OSCC can bring a favorable change in the life of cancer survivors.


Fast nonnegative deconvolution for spike train inference from population calcium imaging.
Fluorescent calcium indicators are becoming increasingly popular as a means for observing the spiking activity of large neuronal populations. Unfortunately, extracting the spike train of each neuron from a raw fluorescence movie is a nontrivial problem. This work presents a fast nonnegative deconvolution filter to infer the approximately most likely spike train of each neuron, given the fluorescence observations. This algorithm outperforms optimal linear deconvolution (Wiener filtering) on both simulated and biological data. The performance gains come from restricting the inferred spike trains to be positive (using an interior-point method), unlike the Wiener filter. The algorithm runs in linear time, and is fast enough that even when simultaneously imaging >100 neurons, inference can be performed on the set of all observed traces faster than real time. Performing optimal spatial filtering on the images further refines the inferred spike train estimates. Importantly, all the parameters required to perform the inference can be estimated using only the fluorescence data, obviating the need to perform joint electrophysiological and imaging calibration experiments.


Metataxonomic and Metagenomic Approaches vs. Culture-Based Techniques for Clinical Pathology.
Diagnoses that are both timely and accurate are critically important for patients with life-threatening or drug resistant infections. Technological improvements in High-Throughput Sequencing (HTS) have led to its use in pathogen detection and its application in clinical diagnoses of infectious diseases. The present study compares two HTS methods, 16S rRNA marker gene sequencing (metataxonomics) and whole metagenomic shotgun sequencing (metagenomics), in their respective abilities to match the same diagnosis as traditional culture methods (culture inference) for patients with ventilator associated pneumonia (VAP). The metagenomic analysis was able to produce the same diagnosis as culture methods at the species-level for five of the six samples, while the metataxonomic analysis was only able to produce results with the same species-level identification as culture for two of the six samples. These results indicate that metagenomic analyses have the accuracy needed for a clinical diagnostic tool, but full integration in diagnostic protocols is contingent on technological improvements to decrease turnaround time and lower costs.


Wound management and the use of mouth rinse in mandibular third molar surgery.
The aim of this survey was to assess the knowledge and practice of Swiss dentists regarding wound management and the use of mouth rinse in surgical removal of mandibular third molars (MTM). A postal survey was conducted among all 3,288 dentists who are members of the Swiss Dental Society (SSO) representing the majority of dentists in Switzerland. The questionnaire consisted of 13 questions with mostly multiple-choice answers. Demographic profile, surgical experience, the use of antibiotics, and wound management, i.e. wound closure and the use of mouth rinse were assessed. The response rate was 55%. Semi-closed (59.1%), closed (19%) and open wound management (11.7%) were applied most often. Semi-closed wound management was preferred in the German-speaking region (67%) and closed wound management was preferred in the French-speaking region (55%). For semi-closed wound healing, drains impregnated with terra-cortril (42%) and iodoform-Vaseline (40%) were used. For closed wound management, most dentists reported leaving the wound to heal with the blood clot only (60.5%). Most dentists (74.5%) prescribed chlorhexidine 0.2% (CHX) mouth rinse when performing MTM surgery and a combination of immediate preoperative and postoperative use was preferred. Semi-closed wound management with drain and CHX mouth rinse is frequently used in Switzerland in the perioperative management in MTM surgery. It is a well-documented procedure leading to favourable outcomes without using any systemic antibiotics. However, wound management techniques differ between the three linguistic regions.


[Head masters' perception of school-based hostility in Alicante, Spain: a qualitative study].
There is growing social concern about school violence but little is known about how this situation is experienced in the schools. The objective of this study was to know the perception that School's head masters have of the school-based hostility, on their causes, how they value current preventive strategies, and their recommendations to develop future preventive interventions. We conducted an exploratory study with 14 semistructured interviews of School's head masters. Interviews were conducted at the Schools between May and June 2003 in 1 independent school, 9 state comprehensive schools, and 4 other comprehensive schools within the city of Alicante. The perception of the head masters is that the prevalence of violence is low, more verbal than physical and within gender. They identify lack of punctuality, absenteeism and lack of interest as forms of hostility. As causes they identified age, family problems, school environment, society, media (TV and video-games) and poor language skills. Preventive methods in use were, on one hand, extra-lessons and transversal contents foreseen in the Education Law and common to all schools, and, on the other hand, ideological and pedagogical contents which were specific of some centres. Recommendations focused in demands for increased economic support and skilled human resources. School-based violence is not perceived as an alarming school issue, nor by its magnitude neither by the immediate causes and students' characteristics to which it is associated. A heavier emphasis is placed on external and environmental causes perceived as much more difficult to confront.


Use of adaptive optics to determine the optimal ocular spherical aberration.
To explore the impact of spherical aberration (SA) on contrast sensitivity using an adaptive optics vision simulator to determine the optimal amount of SA to include in customized corrections of wavefront aberrations. Laboratorio de Optica, Universidad de Murcia, Murcia, Spain, and AMO Groningen BV, Groningen, The Netherlands. An adaptive optics vision simulator consisting of a wavefront sensor, a 97-segmented deformable mirror to induce and correct aberrations of the eye, and a visual testing path was constructed for this study. The deformable mirror allows the effective ocular wavefront aberration to be manipulated and the resulting visual performance to be measured simultaneously. Subjective measurements of contrast sensitivity at 15 cycles per degree were performed with a 4.8 mm pupil in 5 subjects with different levels of naturally occurring SA. Contrast sensitivity was measured when SA values of -0.09 microm, 0.0 microm, 0.09 microm, and 0.182 microm were induced when the other natural aberrations of the eye were present, when the aberrations were corrected, and at defocus values of +/-0.25 diopter (D) and +/-0.50 D. Subjects experienced peak contrast sensitivity performance with varying levels of SA when their natural aberrations were present; however, average contrast performance peaked at 0 mum of SA. When all higher-order aberrations were corrected, all 5 subjects' peak performance occurred at 0 microm of SA. The adaptive optics vision simulator reduced the root-mean-square wavefront aberration of the eye by up to a factor of 4 and allowed noninvasive testing of the visual performance resulting from any ocular wavefront aberration introduced by customized correction procedures. This study showed that, on average, contrast performance peaked when SA was completely corrected.


Adhesion-induced tyrosine phosphorylation of the p130 src substrate.
Adhesion of cells to the extracellular matrix leads to an increase in the tyrosine phosphorylation of a specific set of proteins, three of which have now been identified as the focal adhesion proteins pp125FAK, paxillin and tensin. In addition, we have previously noted the adhesion-induced tyrosine phosphorylation of a fourth protein, with an apparent molecular mass of 130. As in the case of FAK, paxillin and tensin, a 130 kDa protein is also found to be highly tyrosine phosphorylated in Rous sarcoma virus (RSV)-transformed cells. This protein forms a stable complex with pp60src and is directly phosphorylated by activated forms of c-src. Using a monoclonal antibody (mAb 4F4) specific for the src-associated p130 we show that p130 is also phosphorylated in response to cell adhesion. Immunoprecipitation of p130 followed by an anti-phosphotyrosine immunoblot revealed that adhesion of rat embryo fibroblasts (REF52) to fibronectin (FN) led to a significant increase in the phosphotyrosine content of p130. Furthermore, a comparison of cell lysates before and after immunoprecipitation confirmed the absence of tyrosine phosphorylated p130 from lysates immunoprecipitated with mAb 4F4. Immunofluorescence staining of REF52s revealed that p130 is found in focal adhesions as well as along stress fibers in a pattern reminiscent of that exhibited by alpha-actinin. In addition, in many cells, we found significant staining in the nucleus, but evidence is presented that the nuclear staining is not due to tyrosine phosphorylated p130. Finally, unlike pp125FAK, p130 does not appear to be itself a kinase as evidence by immune-complex kinase assays carried out in the presence or absence of exogenous substrates.


Dorsal root ganglia neurons and differentiated adipose-derived stem cells: an in vitro co-culture model to study peripheral nerve regeneration.
Dorsal root ganglia (DRG) neurons, located in the intervertebral foramina of the spinal column, can be used to create an in vitro system facilitating the study of nerve regeneration and myelination. The glial cells of the peripheral nervous system, Schwann cells (SC), are key facilitators of these processes; it is therefore crucial that the interactions of these cellular components are studied together. Direct contact between DRG neurons and glial cells provides additional stimuli sensed by specific membrane receptors, further improving the neuronal response. SC release growth factors and proteins in the culture medium, which enhance neuron survival and stimulate neurite sprouting and extension. However, SC require long proliferation time to be used for tissue engineering applications and the sacrifice of an healthy nerve for their sourcing. Adipose-derived stem cells (ASC) differentiated into SC phenotype are a valid alternative to SC for the set-up of a co-culture model with DRG neurons to study nerve regeneration. The present work presents a detailed and reproducible step-by-step protocol to harvest both DRG neurons and ASC from adult rats; to differentiate ASC towards a SC phenotype; and combines the two cell types in a direct co-culture system to investigate the interplay between neurons and SC in the peripheral nervous system. This tool has great potential in the optimization of tissue-engineered constructs for peripheral nerve repair.


Metabolic Effects Associated with ICS in Patients with COPD and Comorbid Type 2 Diabetes: A Historical Matched Cohort Study.
Management guidelines for chronic obstructive pulmonary disease (COPD) recommend that inhaled corticosteroids (ICS) are prescribed to patients with the most severe symptoms. However, these guidelines have not been widely implemented by physicians, leading to widespread use of ICS in patients with mild-to-moderate COPD. Of particular concern is the potential risk of worsening diabetic control associated with ICS use. Here we investigate whether ICS therapy in patients with COPD and comorbid type 2 diabetes mellitus (T2DM) has a negative impact on diabetic control, and whether these negative effects are dose-dependent. This was a historical matched cohort study utilising primary care medical record data from two large UK databases. We selected patients aged ≥40 years with COPD and T2DM, prescribed ICS (n = 1360) or non-ICS therapy (n = 2642) between 2008 and 2012. The primary endpoint was change in HbA1c between the baseline and outcome periods. After 1:1 matching, each cohort consisted of 682 patients. Over the 12-18-month outcome period, patients prescribed ICS had significantly greater increases in HbA1c values compared with those prescribed non-ICS therapies; adjusted difference 0.16% (95% confidence interval [CI]: 0.05-0.27%) in all COPD patients, and 0.25% (95% CI: 0.10-0.40%) in mild-to-moderate COPD patients. Patients in the ICS cohort also had significantly more diabetes-related general practice visits per year and received more frequent glucose strip prescriptions, compared with those prescribed non-ICS therapies. Patients prescribed higher cumulative doses of ICS (>250 mg) had greater odds of increased HbA1c and/or receiving additional antidiabetic medication, and increased odds of being above the Quality and Outcomes Framework (QOF) target for HbA1c levels, compared with those prescribed lower cumulative doses (≤125 mg). For patients with COPD and comorbid T2DM, ICS therapy may have a negative impact on diabetes control. Patients prescribed higher cumulative doses of ICS may be at greater risk of diabetes progression. ENCePP ENCEPP/SDPP/6804.


The time course of interocular suppression in normal and amblyopic subjects.
The authors measured the time course of interocular suppression of five normal subjects and eleven patients with amblyopia (strabismic and/or anisometropic). Orthogonal gratings were presented dichoptically for durations that ranged from 10-6000 msec. All normal observers reported fusion or superimposition of the orthogonal gratings for short stimuli and reported binocular rivalry for stimuli longer than 150 msec. At long presentation times, all amblyopes constantly suppressed the pattern that was presented to their amblyopic eye. Six amblyopes showed superimposition of the two patterns at short presentation times. Of these, three had time courses similar to those of normal observers; the other three had a much shorter onset of suppression (about 80 msec). The remaining five amblyopes perceived only the pattern of the dominant eye at short stimulus durations; at intermediate durations, they reported partial superimposition of the stimuli, whereas at the longest stimulus durations, again only the stimulus of the dominant eye was perceived. The results suggest that binocular rivalry in normal observers and strabismic suppression in amblyopes are mediated by different mechanisms. The heterogeneity of the time courses of suppression in amblyopes might result from differences in the disturbances of early visual development (age at onset of strabismus and/or anisometropia, origin, and therapy).


The effects of familiarity and social hierarchy on group membership decisions in a social fish.
Members of animal groups face a trade-off between the benefits of remaining with a familiar group and the potential benefits of dispersing into a new group. Here, we examined the group membership decisions of Neolamprologus pulcher, a group-living cichlid. We found that subordinate helpers showed a preference for joining familiar groups, but when choosing between two unfamiliar groups, helpers did not preferentially join groups that maximized their social rank. Rather, helpers preferred groups containing larger, more dominant individuals, despite receiving significantly more aggression within these groups, possibly owing to increased protection from predation in such groups. These results suggest a complex decision process in N. pulcher when choosing among groups, dependent not only on familiarity but also on the social and life-history consequences of joining new groups.


A randomised, blinded, trial of clopidogrel versus aspirin in patients at risk of ischaemic events (CAPRIE). CAPRIE Steering Committee.
Many clinical trials have evaluated the benefit of long-term use of antiplatelet drugs in reducing the risk of clinical thrombotic events. Aspirin and ticlopidine have been shown to be effective, but both have potentially serious adverse effects. Clopidogrel, a new thienopyridine derivative similar to ticlopidine, is an inhibitor of platelet aggregation induced by adenosine diphosphate. CAPRIE was a randomised, blinded, international trial designed to assess the relative efficacy of clopidogrel (75 mg once daily) and aspirin (325 mg once daily) in reducing the risk of a composite outcome cluster of ischaemic stroke, myocardial infarction, or vascular death; their relative safety was also assessed. The population studied comprised subgroups of patients with atherosclerotic vascular disease manifested as either recent ischaemic stroke, recent myocardial infarction, or symptomatic peripheral arterial disease. Patients were followed for 1 to 3 years. 19,185 patients, with more than 6300 in each of the clinical subgroups, were recruited over 3 years, with a mean follow-up of 1.91 years. There were 1960 first events included in the outcome cluster on which an intention-to-treat analysis showed that patients treated with clopidogrel had an annual 5.32% risk of ischaemic stroke, myocardial infarction, or vascular death compared with 5.83% with aspirin. These rates reflect a statistically significant (p = 0.043) relative-risk reduction of 8.7% in favour of clopidogrel (95% Cl 0.3-16.5). Corresponding on-treatment analysis yielded a relative-risk reduction of 9.4%. There were no major differences in terms of safety. Reported adverse experiences in the clopidogrel and aspirin groups judged to be severe included rash (0.26% vs 0.10%), diarrhoea (0.23% vs 0.11%), upper gastrointestinal discomfort (0.97% vs 1.22%), intracranial haemorrhage (0.33% vs 0.47%), and gastrointestinal haemorrhage (0.52% vs 0.72%), respectively. There were ten (0.10%) patients in the clopidogrel group with significant reductions in neutrophils (< 1.2 x 10(9)/L) and 16 (0.17%) in the aspirin group. Long-term administration of clopidogrel to patients with atherosclerotic vascular disease is more effective than aspirin in reducing the combined risk of ischaemic stroke, myocardial infarction, or vascular death. The overall safety profile of clopidogrel is at least as good as that of medium-dose aspirin.


Photoacoustic holographic imaging of absorbers embedded in silicone.
Light absorbing objects embedded in silicone have been imaged using photoacoustic digital holography. The photoacoustic waves were generated using a pulsed Nd:YAG laser, λ=1064 nm, and pulse length=12 ns. When the waves reached the silicone surface, they were measured optically along a line using a scanning laser vibrometer. The acoustic waves were then digitally reconstructed using a holographic algorithm. The laser vibrometer is proven to be sensitive enough to measure the surface velocity due to photoacoustic waves generated from laser pulses with a fluence allowed for human tissue. It is also shown that combining digital holographic reconstructions for different acoustic wavelengths provides images with suppressed noise and improved depth resolution. The objects are imaged at a depth of 16.5 mm with a depth resolution of 0.5 mm.


Suicide in France: some hypotheses.
The relatively high suicide rate in France is investigated and a number of influences are hypothesized as causative. These include, on a societal and demographic level, a history of high immigration, low emigration, a high proportion of old people, high urbanization, extraordinarily high alcoholism, and the extreme gap in income between upper and lower classes. The rigid bureaucracy of the state can leave the individual feeling infuriated and defeated. The legal system produces many injustices, such as long imprisonment without charges. The Church is liberal and supplies little prophylaxis against suicide. Overall, social integration must be judged to be low. Culturally, French values include an underlying pessimism, no strong fear of death, strong pressures to behave correctly and much malice toward neighbors. The modal personality structure contains defensive, constricted elements producing a vulnerable pseudo-autonomy. French child-rearing practices are effective in producing such personalities.


Ring lithiation of 1,8-bis(dimethylamino)naphthalene: another side of the 'proton sponge coin'.
It has been found that 1,8-bis(dimethylamino)naphthalene (DMAN), unlike N,N-dimethylaniline, undergoes ring metallation in the n-BuLi-TMEDA-Et2O system with a low selectivity and in poor total yields. The situation is significantly improved in the t-BuLi-TMEDA-n-hexane system when 3- and 4-lithium derivatives become the only reaction products obtained in good yields. The formation of 3-Li-DMAN is especially desired since no method of direct meta-functionalization of DMAN is known to date. The relative stability and structure of DMAN lithium derivatives have been examined with the help of X-ray and multinuclear NMR measurements as well as DFT calculations.


New insights into autophagic cell death in the gypsy moth Lymantria dispar: a proteomic approach.
Autophagy is an evolutionary ancient process based on the activity of genes conserved from yeast to metazoan taxa. Whereas its role as a mechanism to provide energy during cell starvation is commonly accepted, debate continues about the occurrence of autophagy as a means specifically activated to achieve cell death. The IPLB-LdFB insect cell line, derived from the larval fat body of the lepidoptera Lymantria dispar, represents a suitable model to address this question, as both autophagic and apoptotic cell death can be induced by various stimuli. Using morphological and functional approaches, we have observed that the culture medium conditioned by IPLB-LdFB cells committed to death by the ATPase inhibitor oligomycin A stimulates autophagic cell death in untreated IPLB-LdFB cells. Moreover, proteomic analysis of the conditioned media suggests that, in IPLB-LdFB cells, oligomycin A promotes a shift towards lipid metabolism, increases oxidative stress and specifically directs the cells towards autophagic activity.


Simulated patients' perspectives of and perceived role in medical students' professional identity development.
Much has been written about medical students' professional identity formation, the process of "becoming" a doctor. During their training, medical students interact with a range of teachers and trainers. Among these are simulated patients (SPs) who role-play patients, assisting students with their communication, procedural, and physical examination skills. With SPs regularly interacting with students, this qualitative study explored their views of students' emerging professional identities at one Australian medical school. SPs' contributions to developing professional identities were also explored. Fourteen SPs were interviewed individually or in pairs. After template analysis of the transcripts using a priori themes, a follow-up focus group (n = 7) was arranged. Although being older (implying maturity and more life experience) and exposure to real patients and previous health care experience were identified as contributing to developing an identity as a doctor, SPs recognized that for some, an existing professional identity might impede the development of a new identity. Simulated patients were of the opinion that they contributed to students' professional identities by creating a supportive environment for honing skills, which they did by realistically role-playing patient scripts, by making their bodies available, and by providing feedback as "patients." Through their authentic portrayal of patients and through their feedback, we are of the opinion that our SPs can contribute to students' developing identities as doctors. As lay individuals who often encounter students longitudinally, we believe that SPs offer a particular lens through which to view students' emerging identities as future doctors.


Barcode UK: A complete DNA barcoding resource for the flowering plants and conifers of the United Kingdom.
DNA barcoding and metabarcoding provide new avenues for investigating biological systems. These techniques require well-curated reference libraries with extensive coverage. Generating an exhaustive national DNA barcode reference library can open up new avenues of research in ecology, evolution and conservation, yet few studies to date have created such a resource. In plant DNA barcoding, herbarium collections provide taxonomically robust material but also pose challenges in lab processing. Here, we present a national DNA barcoding resource covering all of the native flowering plants and conifers of the United Kingdom. This represents 1,482 plant species, with the majority of specimens (81%) sourced from herbaria. Using Sanger sequencing of the plant DNA barcode markers, rbcL, matK, and ITS2, at least one DNA barcode was retrieved from 98% of the UK flora. We sampled from multiple individuals, resulting in a species coverage for rbcL of 96% (4,477 sequences), 90% for matK (3,259 sequences) and 75% for ITS2 (2,585 sequences). Sequence recovery was lower for herbarium material compared to fresh collections, with the age of the specimen having a significant effect on the success of sequence recovery. Species level discrimination was highest with ITS2, however, the ability to successfully retrieve a sequence was lowest for this region. Analyses of the genetic distinctiveness of species across a complete flora showed DNA barcoding to be informative for all but the most taxonomically complex groups. The UK flora DNA barcode reference library provides an important resource for many applications that require plant identification from DNA.


Surgical VLS Therapy of Oesophageal Achalasia in Pediatric Age: Four Case Reports.
The Authors present a retrospective review of their record of cases, characterized by 4 cases of achalasia in which it was performed a Heller myotomy with front fundoplication (Thall) in laparoscopic approach in the period from 2012 to 2019. In paediatric achalasia, the laparoscopic Heller myotomy seems to be the best treatment because of its multiple advantages offered by the minimally invasive technique. First of all, thanks to the video-technique, which allows a complete and extended myotomy, the accuracy of this operation is maximized; moreover, the post-operative pain is widely reduced, thanks to the minimal dissection and traction of the tissues; finally, but not negligible, this approach ensures a better aesthetic result than the classic open technique. With regard to the front fundoplication, the Authors suggest that it is mandatory because, even if it extends the operating time, it ensures a natural protection to the myotomy herniated mucosa and avoids gastro-oesophageal reflux, which often occurs after the surgical correction, thus obliging to perform a reoperation.


Genome-Scale Assessment of Age-Related DNA Methylation Changes in Mouse Spermatozoa.
DNA methylation plays important roles in the production and functioning of spermatozoa. Recent studies have suggested that DNA methylation patterns in spermatozoa can change with age, but the regions susceptible to age-related methylation changes remain to be fully elucidated. In this study, we conducted genome-scale DNA methylation profiling of spermatozoa obtained from C57BL/6N mice at 8 weeks (8w), 18 weeks (18w) and 17 months of age (17m). There was no substantial difference in the global DNA methylation patterns between 18w and 17m samples except for a slight increase of methylation levels in long interspersed nuclear elements in the 17m samples. We found that maternally methylated imprinting control regions (mICRs) and spermatogenesis-related gene promoters had 5-10% higher methylation levels in 8w samples than in 18w or 17m samples. Analysis of individual sequence reads suggested that these regions were fully methylated (80-100%) in a subset of 8w spermatozoa. These regions are also known to be highly methylated in a subset of postnatal spermatogonia, which might be the source of the increased DNA methylation in 8w spermatozoa. Another possible source was contamination by somatic cells. Although we carefully purified the spermatozoa, it was difficult to completely exclude the possibility of somatic cell contamination. Further studies are needed to clarify the source of the small increase in DNA methylation in the 8w samples. Overall, our findings suggest that DNA methylation patterns in mouse spermatozoa are relatively stable throughout reproductive life.