Clinical Pharmacists as Educators in Family Medicine Residency Programs: A CERA Study of Program Directors.
The clinical pharmacist's role within family medicine residency programs (FMRPs) is well established. However, there is limited information regarding perceptions of program directors (PDs) about clinical pharmacy educators. The study objectives were (1) to estimate the prevalence of clinical pharmacists within FMRPs and (2) to determine barriers and motivations for incorporation of clinical pharmacists as educators. The Council of Academic Family Medicine Educational Research Alliance (CERA) distributed an electronic survey to PDs. Questions addressed formalized pharmacotherapy education, clinical pharmacists in educator roles, and barriers and benefits of clinical pharmacists in FMRPs. The overall response rate was 50% (224/451). Seventy-six percent (170/224) of the responding PDs reported that clinical pharmacists provide pharmacotherapy education in their FMRPs, and 57% (97/170) consider clinical pharmacists as faculty members. In programs with clinical pharmacists, 72% (83/116) of PDs reported having a systematic approach for teaching pharmacotherapy versus 22% (21/95) in programs without. In programs without clinical pharmacists, the top barrier to incorporation was limited ability to bill for clinical services 48% (43/89) versus 29% (32/112) in programs with clinical pharmacists. In both programs with and without clinical pharmacists, the top benefit of having clinical pharmacists was providing a collaborative approach to pharmacotherapy education for residents (35% and 36%, respectively). Less than half of FMRPs incorporate clinical pharmacists as faculty members. Despite providing collaborative approaches to pharmacotherapy education, their limited ability to bill for services is a major barrier.


Why and how to step down chronic asthma drugs.
Asthma is a common chronic airways disease. The goal of asthma management is to control symptoms while minimizing the side effects of treatment. Following a period of stable asthma, clinicians should consider stepping down treatment. This approach is recommended by current guidelines. Step-down has been studied for several types of asthma drug regimens, and certain approaches may have lower risk than others. Systematic reviews of multiple trials support the following specific step-down approaches: optimizing inhaled corticosteroid dosing when stepping down oral corticosteroid, reducing inhaled corticosteroid from a higher dose, lowering inhaled corticosteroid-long acting bronchodilator (ICS-LABA) dose while adding ICS-LABA on-demand, adding leukotriene receptor antagonist (LTRA) while lowering inhaled corticosteroid dose, and using allergen immunotherapy when reducing inhaled corticosteroid from a higher dose. Systematic reviews of multiple trials support an increased risk of asthma exacerbation for patients who completely stop taking inhaled corticosteroid or long acting bronchodilator. Strategies to implement step-down in practice include the use of risk prediction as well as tools to support shared decision making and communication about risk between clinicians and patients.


Phenotypic Similarities Between Late-Onset Autosomal Dominant and Sporadic Alzheimer Disease: A Single-Family Case-Control Study.
The amyloid hypothesis posits that disrupted β-amyloid homeostasis initiates the pathological process resulting in Alzheimer disease (AD). Autosomal dominant AD (ADAD) has an early symptomatic onset and is caused by single-gene mutations that result in overproduction of β-amyloid 42. To the extent that sporadic late-onset AD (LOAD) also results from dysregulated β-amyloid 42, the clinical phenotypes of ADAD and LOAD should be similar when controlling for the effects of age. To use a family with late-onset ADAD caused by a presenilin 1 (PSEN1) gene mutation to mitigate the potential confound of age when comparing ADAD and LOAD. This case-control study was conducted at the Knight Alzheimer Disease Research Center at Washington University, St Louis, Missouri, and other National Institutes of Aging-funded AD centers in the United States. Ten PSEN1 A79V mutation carriers from multiple generations of a family with late-onset ADAD and 12 noncarrier family members were followed up at the Knight Alzheimer Disease Research Center (1985-2015) and 1115 individuals with neuropathologically confirmed LOAD were included from the National Alzheimer Coordinating Center database (September 2005-December 2014). Data analysis was completed in January 2016, including Knight Alzheimer Disease Research Center patient data collected up until the end of 2015. Planned comparison of clinical characteristics between cohorts, including age at symptom onset, associated symptoms and signs, rates of progression, and disease duration. Of the PSEN1 A79V carriers in the family with late-onset ADAD, 4 were female (57%); among those with LOAD, 529 were female (47%). Seven mutation carriers (70%) developed AD dementia, while 3 were yet asymptomatic in their seventh and eighth decades of life. No differences were observed between mutation carriers and individuals with LOAD concerning age at symptom onset (mutation carriers: mean, 75 years [range, 63-77 years] vs those with LOAD: mean, 74 years [range, 60-101 years]; P = .29), presenting symptoms (memory loss in 7 of 7 mutation carriers [100%] vs 958 of 1063 individuals with LOAD [90.1%]; P ≥ .99) and duration (mutation carriers: mean, 9.9 years [range, 2.3-12.8 years] vs those with LOAD: 9 years [range, 1-27 years]; P = .73), and rate of progression of dementia (median annualized change in Clinical Dementia Rating-Sum of Boxes score, mutation carriers: 1.2 [range, 0.1-3.3] vs those with LOAD: 1.9 [range, -3.5 to 11.9]; P = .73). Early emergence of comorbid hallucinations and delusions were observed in 57% of individuals with ADAD (4 of 7) vs 19% of individuals with LOAD (137 of 706) (P = .03). Three of 12 noncarriers (25%) from the PSEN1 A79V family are potential phenocopies as they also developed AD dementia (median age at onset, 76.0 years). In this family, the amyloidogenic PSEN1 A79V mutation recapitulates the clinical attributes of LOAD. Previously reported clinical phenotypic differences between individuals with ADAD and LOAD may reflect age- or mutation-dependent effects.


Compendium of measures to prevent disease associated with animals in public settings, 2005.
Multiple venues encourage or permit the public to come in contact with animals, resulting in millions of human-animal contacts each year. These settings include county or state fairs, petting zoos, animal swap meets, pet stores, zoologic institutions, circuses, carnivals, farm tours, livestock-birthing exhibits, educational exhibits at schools, and wildlife photo opportunities. Although multiple benefits of human-animal contact exist, infectious diseases, rabies exposures, injuries, and other human health problems associated with these settings are of concern. Rabid or potentially rabid animals in public settings can result in extensive public health investigation and action. Infectious disease outbreaks reported during the previous decade have been attributed to multiple organisms, including Escherichia coli O157:H7, Salmonella, Coxiella burnetti, Mycobacterium tuberculosis, and ringworm. Such incidents have substantial medical, public health, legal, and economic effects. This report provides standardized recommendations for public health officials, veterinarians, animal venue operators, animal exhibitors, visitors to animal venues and exhibits, and others concerned with disease-control and with minimizing risks associated with animals in public settings. The recommendation to wash hands is the single most important prevention step for reducing the risk for disease transmission. Other critical recommendations are that venues include transition areas between animal areas and nonanimal areas (where food is sold) and that animals are properly cared for and managed in public settings. In addition, this report recommends educating venue operators, staff, exhibitors, and visitors regarding the risk for disease transmission where animal contact is possible.


Metacarpophalangeal joint loads during bonobo locomotion: model predictions versus proxies.
The analysis of internal trabecular and cortical bone has been an informative tool for drawing inferences about behaviour in extant and fossil primate taxa. Within the hand, metacarpal bone architecture has been shown to correlate well with primate locomotion; however, the extent of morphological differences across taxa is unexpectedly small given the variability in hand use. One explanation for this observation is that the activity-related differences in the joint loads acting on the bone are simply smaller than estimated based on commonly used proxies (i.e. external loading and joint posture), which neglect the influence of muscle forces. In this study, experimental data and a musculoskeletal finger model are used to test this hypothesis by comparing differences between climbing and knuckle-walking locomotion of captive bonobos (Pan paniscus) based on (i) joint load magnitude and direction predicted by the models and (ii) proxy estimations. The results showed that the activity-related differences in predicted joint loads are indeed much smaller than the proxies would suggest, with joint load magnitudes being almost identical between the two locomotor modes. Differences in joint load directions were smaller but still evident, indicating that joint load directions might be a more robust indicator of variation in hand use than joint load magnitudes. Overall, this study emphasizes the importance of including muscular forces in the interpretation of skeletal remains and promotes the use of musculoskeletal models for correct functional interpretations.


[Ectopic mediastinal parathyroid adenoma. Detection with a radioisotopic probe and resolution with videothoracoscopy].
We report a 66 years old woman with a diagnosis of primary hyperparathyroidism. Localization to mediastinum was obtained with parathyroid scintigraphy using 99mtc-methoxy-isobutyl-isonitrite (Tc99-MIBI). The patient was successfully operated upon by means of a videothora-coscopic approach. During the procedure serum parathormone was measured before and 10 minutes after adenomectomy, showing a more than 50% reduction from the basal level. An attempt to detect the precise site of the adenoma with a Tc99-MIBI probe was unsuccessful because of its proximity to the myocardium, but radioactivity was confirmed on the surgical specimen after resection. The patient's calcemia and parathormone levels became normal during the postoperative course and she remains normocalcemic 9 months after the procedure. In our case, preoperative localization and intraoperative parathormone measurements were both very useful for confirming surgical success; the intraoperative localization with a radioactive probe was not useful, but radioactivity was confirmed after resection on the surgical specimen. The endoscopic surgical procedure with videothoracoscopy was well tolerated, less painful than a thoracotomy, and it shortened the hospitalization period.


Exercise Counteracts Aging-Related Memory Impairment: A Potential Role for the Astrocytic Metabolic Shuttle.
Age-related cognitive impairment has become one of the most common health threats in many countries. The biological substrate of cognition is the interconnection of neurons to form complex information processing networks. Experience-based alterations in the activities of these information processing networks lead to neuroadaptation, which is physically represented at the cellular level as synaptic plasticity. Although synaptic plasticity is known to be affected by aging, the underlying molecular mechanisms are not well described. Astrocytes, a glial cell type that is infrequently investigated in cognitive science, have emerged as energy suppliers which are necessary for meeting the abundant energy demand resulting from glutamatergic synaptic activity. Moreover, the concerted action of an astrocyte-neuron metabolic shuttle is essential for cognitive function; whereas, energetic incoordination between astrocytes and neurons may contribute to cognitive impairment. Whether altered function of the astrocyte-neuron metabolic shuttle links aging to reduced synaptic plasticity is unexplored. However, accumulated evidence documents significant beneficial effects of long-term, regular exercise on cognition and synaptic plasticity. Furthermore, exercise increases the effectiveness of astrocyte-neuron metabolic shuttle by upregulation of astrocytic lactate transporter levels. This review summarizes previous findings related to the neuronal activity-dependent astrocyte-neuron metabolic shuttle. Moreover, we discuss how aging and exercise may shape the astrocyte-neuron metabolic shuttle in cognition-associated brain areas.


Sen1 is a key regulator of transcription-driven conflicts.
Cellular homeostasis requires the coordination of several machineries concurrently engaged in the DNA. Wide-spread transcription can interfere with other processes, and transcription-replication conflicts (TRCs) threaten genome stability. The conserved Sen1 helicase not only terminates non-coding transcription but also interacts with the replisome and reportedly resolves genotoxic R-loops. Sen1 prevents genomic instability, but how this relates to its molecular functions remains unclear. We generated high-resolution, genome-wide maps of transcription-dependent conflicts and R-loops using a Sen1 mutant that has lost interaction with the replisome but is termination proficient. We show that, under physiological conditions, Sen1 removes RNA polymerase II at TRCs within genes and the rDNA and at sites of transcription-transcription conflicts, thus qualifying as a "key regulator of conflicts." We demonstrate that genomic stability is affected by Sen1 mutation only when in addition to its role at the replisome, the termination of non-coding transcription or R-loop removal are additionally compromised.


The healthiness and sustainability of national and global food based dietary guidelines: modelling study.
To analyse the health and environmental implications of adopting national food based dietary guidelines (FBDGs) at a national level and compared with global health and environmental targets. Modelling study. 85 countries. Population of 85 countries. A graded coding method was developed and used to extract quantitative recommendations from 85 FBDGs. The health and environmental impacts of these guidelines were assessed by using a comparative risk assessment of deaths from chronic diseases and a set of country specific environmental footprints for greenhouse gas emissions, freshwater use, cropland use, and fertiliser application. For comparison, the impacts of adopting the global dietary recommendations of the World Health Organization and the EAT-Lancet Commission on Healthy Diets from Sustainable Food Systems were also analysed. Each guideline's health and sustainability implications were assessed by modelling its adoption at both the national level and globally, and comparing the impacts to global health and environmental targets, including the Action Agenda on Non-Communicable Diseases, the Paris Climate Agreement, the Aichi biodiversity targets related to land use, and the sustainable development goals and planetary boundaries related to freshwater use and fertiliser application. Adoption of national FBDGs was associated with reductions in premature mortality of 15% on average (95% uncertainty interval 13% to 16%) and mixed changes in environmental resource demand, including a reduction in greenhouse gas emissions of 13% on average (regional range -34% to 35%). When universally adopted globally, most of the national guidelines (83, 98%) were not compatible with at least one of the global health and environmental targets. About a third of the FBDGs (29, 34%) were incompatible with the agenda on non-communicable diseases, and most (57 to 74, 67% to 87%) were incompatible with the Paris Climate Agreement and other environmental targets. In comparison, adoption of the WHO recommendations was associated with similar health and environmental changes, whereas adoption of the EAT-Lancet recommendations was associated with 34% greater reductions in premature mortality, more than three times greater reductions in greenhouse gas emissions, and general attainment of the global health and environmental targets. As an example, the FBDGs of the UK, US, and China were incompatible with the climate change, land use, freshwater, and nitrogen targets, and adopting guidelines in line with the EAT-Lancet recommendation could increase the number of avoided deaths from 78 000 (74 000 to 81 000) to 104 000 (96 000 to 112 000) in the UK, from 480 000 (445 000 to 516 000) to 585 000 (523 000 to 646 000) in the USA, and from 1 149 000 (1 095 000 to 1 204 000) to 1 802 000 (1 664 000 to 1 941 000) in China. This analysis suggests that national guidelines could be both healthier and more sustainable. Providing clearer advice on limiting in most contexts the consumption of animal source foods, in particular beef and dairy, was found to have the greatest potential for increasing the environmental sustainability of dietary guidelines, whereas increasing the intake of whole grains, fruits and vegetables, nuts and seeds, and legumes, reducing the intake of red and processed meat, and highlighting the importance of attaining balanced energy intake and weight levels were associated with most of the additional health benefits. The health results were based on observational data and assuming a causal relation between dietary risk factors and health outcomes. The certainty of evidence for these relations is mostly graded as moderate in existing meta-analyses.


Rare variants of ATM and risk for Hodgkin's disease and radiation-associated breast cancers.
In this study, we first sought to evaluate whether individuals heterozygous for ATM mutations may have an increased susceptibility to radiation-induced breast cancer (BC) after treatment for Hodgkin's disease (HD). We next sought to determine the frequency of ATM variants in patients with Hodgkin's lymphoma, regardless of coexisting BC, compared with healthy volunteers. Full sequence analysis of ATM was performed on cDNA from peripheral blood lymphocytes from 37 cases of BC after therapeutic radiation therapy for HD and 27 comparison cases with HD and no BC treated during the same time period. The frequency of ATM variants was analyzed in the total group of 64 cases of HD and compared to allele frequencies in 128 ethnically matched controls from the same geographical region. No protein-truncating ATM mutations were observed in cases with HD with or without BC. Missense mutations were more frequent in the cohort with HD compared with patients with BC following HD (P = 0.02). The median time from HD to the development of BC was 18 years in patients with ATM variants compared with 16 years in those with no ATM variants (P = 0.04). Multiple ATM variants, including one homozygous mutation, were observed in 9 HD cases. Heterozygous protein-truncating or missense mutations of ATM were not associated with increased radiation-associated risk of BC after HD. The observation of multiple germ-line mutations and a homozygote suggests that rare ATM variants may constitute cancer-susceptibility alleles in a subset of cases.


Life-history traits of the common snook Centropomus undecimalis in a Caribbean estuary and large-scale biogeographic patterns relevant to management.
The ecology of common snook Centropomus undecimalis in Amatique Bay, a tropical estuary in eastern Guatemala, was investigated and life-history traits were used to conduct a meta-analysis of the species from Florida to Brazil. The reproduction cycle of C. undecimalis in Amatique was strongly related to the precipitation cycle, with a lag of 2 months. Spawning occurred from April to November with a peak spawning after the onset of the summer rains. Protandric sex reversal occurred early in the dry season (December) before somatic recovery from spawning. The growth cycle preceded that of body condition by c. 1 month, and was out of phase with the reproductive cycle. Growth was fast, as many individuals reached >70% of the maximum observed total length (LT , 102 cm) after 3 years. Sex transition occurred within a relatively narrow LT range (70-79 cm), but over a wide range of ages, indicating plasticity in this respect. The meta-analysis indicated a latitudinal-temperature gradient in life-history traits, as well as different seasonal patterns relative to temperature and hydrographical cycles. Centropomus undecimalis from cooler winter waters (e.g. Florida) reach larger maximum LT and LT at sex change, as well as greater gonado-somatic indices and longer life spans. Further, increased fishing mortality results in younger age at sex reversal and male predominance in the populations compared. Recognition of large-scale biogeographic patterns in this important, but little studied, fish species helps in the formulation of management advice in other areas of its occurrence.


Identifying key regulatory genes in the whole blood of septic patients to monitor underlying immune dysfunctions.
There is currently no reliable tool available to measure immune dysfunction in septic patients in the clinical setting. This proof-of-concept study assesses the potential of gene expression profiling of whole blood as a tool to monitor immune dysfunction in critically ill septic patients. Whole-blood samples were collected daily for up to 5 days from patients admitted to the intensive care unit with sepsis. RNA isolated from whole-blood samples was assayed on Illumina HT-12 gene expression microarrays consisting of 48,804 probes. Microarray analysis identified 3,677 genes as differentially expressed across 5 days between septic patients and healthy controls. Of the 3,677 genes, biological pathway analysis identified 86 genes significantly downregulated in the sepsis patients were present in pathways relating to immune response. These 86 genes correspond to known immune pathways implicated in sepsis, including lymphocyte depletion, reduced T-lymphocyte activation, and deficient antigen presentation. Furthermore, expression levels of these genes correlated with clinical severity, with a significantly greater degree of downregulation found in nonsurvivors compared with survivors. The results show that whole-blood gene expression analysis can capture systemic immune dysfunctions in septic patients. Our study provides an experimental basis to support further study on the use of a gene expression-based assay, to assess immunosuppression, and to guide immunotherapy in future clinical trials.


Caring while living apart.
In recent years, researchers have paid increasing attention to the issue of care in the new types of family and partner relationships. The aim of this study is to examine expectations and attitudes concerning care among elderly people in LAT (Living Apart Together)- relationships in Sweden, that is, people who have a long-term intimate relationship, but who do not share a common home. Questionnaires were completed by 116 elderly people in LAT-relationships. Partners are ranked as the main providers of care, especially by men, but there is considerable variation in the answers. This seems to be principally related to the degree of flexibility in this type of relationship. Some individuals see a LAT-relationship as a marriage-like relationship, while others see it primarily as a type of relationship that guarantees them the possibility of maintaining their own independent way of life.


Double Monoubiquitination Modifies the Molecular Recognition Properties of p15PAF Promoting Binding to the Reader Module of Dnmt1.
The proliferating cell nuclear antigen (PCNA)-associated factor p15PAF is a nuclear protein that acts as a regulator of DNA repair during DNA replication. The p15PAF gene is overexpressed in several types of human cancer, and its function is regulated by monoubiquitination of two lysines (K15 and K24) at the protein N-terminal region. We have previously shown that p15PAF is an intrinsically disordered protein which partially folds upon binding to PCNA and independently contacts DNA through its N-terminal tail. Here we present an NMR conformational characterization of p15PAF monoubiquitinated at both K15 and K24 via a disulfide bridge mimicking the isopeptide bond. We show that doubly monoubiquitinated p15PAF is monomeric, intrinsically disordered, and binds to PCNA as nonubiquitinated p15PAF does but interacts with DNA with reduced affinity. Our SAXS-derived conformational ensemble of doubly monoubiquitinated p15PAF shows that the ubiquitin moieties, separated by eight disordered residues, form transient dimers because of the high local effective ubiquitin concentration. This observation and the sequence similarity with histone H3 N-terminal tail suggest that doubly monoubiquitinated p15PAF is a binding target of DNA methyl transferase Dnmt1, as confirmed by calorimetry. Therefore, doubly monoubiquitinated p15PAF directly interacts with PCNA and recruits Dnmt1 for maintenance of DNA methylation during replication.


Molecular and functional characterization of the non-isopeptide-selective ETB receptor in endothelial cells. Receptor coupling to nitric oxide synthase.
There is accumulating evidence that endothelial cells express a non-isopeptide-selective endothelin (ET) receptor, ETB, which may be responsible for ET-1-induced transient vasorelaxation. The purpose of the present study was to seek direct evidence for ETB receptor expression in human umbilical vein endothelial cells (HUVEC) and to characterize its functional role in HUVEC and in Chinese hamster ovary cells stably transfected with ETB receptor cDNA (CHO-ETB). Reverse polymerase chain reaction using HUVEC total RNA and ETB receptor-specific oligonucleotide primers firmly demonstrated the presence of an endogenous transcript of the appropriate molecular size. Next, a biotinylated ligand specifically recognizing the ETB receptor, IRL-1620, was synthesized, and immunocytochemical mapping of binding sites was performed in CHO-ETB cells. Specific binding of biotinylated IRL-1620 was evident in CHO-ETB cells, confirming appropriate cell surface receptor expression. Continuous nitric oxide (NO) monitoring with NO-selective electrode revealed a dose-dependent ET-1 stimulation of NO production by HUVEC. Stable transfection of CHO-ETB cells with endothelial nitric oxide synthase (NOS), but not mock-transfection, imparted responsiveness to ET-1 similar to that for HUVEC and was characterized by the immediate release of NO. Protein tyrosine kinase-dependent and calcium-calmodulin-dependent pathways were involved in ET-1-induced activation of the constitutive NOS in CHO-ETB/NOS cells, but coupling of the receptor to the enzyme in HUVEC appeared to be predominantly protein tyrosine kinase-dependent. Although sufficient, calcium/calmodulin system was not an obligatory prerequisite for the ET-1-induced activation of NOS in HUVEC. In conclusion, using two cell systems, we demonstrated that the ETB receptor is functionally coupled to NOS and coordinates the generation of NO via a tyrosine kinase-dependent and a calcium/calmodulin-dependent pathway.


Application of protein engineering to enhance crystallizability and improve crystal properties.
Until recently, protein crystallization has mostly been regarded as a stochastic event over which the investigator has little or no control. With the dramatic technological advances in synchrotron-radiation sources and detectors and the equally impressive progress in crystallographic software, including automated model building and validation, crystallization has increasingly become the rate-limiting step in X-ray diffraction studies of macromolecules. However, with the advent of recombinant methods it has also become possible to engineer target proteins and their complexes for higher propensity to form crystals with desirable X-ray diffraction qualities. As most proteins that are under investigation today are obtained by heterologous overexpression, these techniques hold the promise of becoming routine tools with the potential to transform classical crystallization screening into a more rational high-success-rate approach. This article presents an overview of protein-engineering methods designed to enhance crystallizability and discusses a number of examples of their successful application.


Is the tobacco control movement misrepresenting the acute cardiovascular health effects of secondhand smoke exposure? An analysis of the scientific evidence and commentary on the implications for tobacco control and public health practice.
While chronic exposure to secondhand smoke has been well recognized as a cause of heart disease in nonsmokers, there has been recent speculation about the potential acute cardiovascular effects of transient exposure to secondhand smoke among nonsmokers; in particular, the possibility that such exposure could increase the risk of acute myocardial infarction even in an otherwise healthy nonsmoker. This paper reviews the claims being made by a number of anti-smoking and public health groups regarding the acute cardiovascular effects of secondhand smoke exposure among otherwise healthy adults, analyzes the validity of these claims based on a review of the scientific evidence, and discusses the implications of the findings for tobacco control and public health practice. Based on the analysis, it appears that a large number of anti-smoking organizations are making inaccurate claims that a single, acute, transient exposure to secondhand smoke can cause severe and even fatal cardiovascular events in healthy nonsmokers. The dissemination of inaccurate information by anti-smoking groups to the public in support of smoking bans is unfortunate because it may harm the tobacco control movement by undermining its credibility, reputation, and effectiveness. Disseminating inaccurate information also represents a violation of basic ethical principles that are a core value of public health practice that cannot and should not be sacrificed, even for a noble end such as protecting nonsmokers from secondhand smoke exposure. How the tobacco control movement responds to this crisis of credibility will go a long way towards determining the future effectiveness of the movement and its ability to continue to save lives and protect the public's health.


Mitochondrial damage-associated inflammation highlights biomarkers in PRKN/PINK1 parkinsonism.
There is increasing evidence for a role of inflammation in Parkinson's disease. Recent research in murine models suggests that parkin and PINK1 deficiency leads to impaired mitophagy, which causes the release of mitochondrial DNA (mtDNA), thereby triggering inflammation. Specifically, the CGAS (cyclic GMP-AMP synthase)-STING (stimulator of interferon genes) pathway mitigates activation of the innate immune system, quantifiable as increased interleukin-6 (IL6) levels. However, the role of IL6 and circulating cell-free mtDNA in unaffected and affected individuals harbouring mutations in PRKN/PINK1 and idiopathic Parkinson's disease patients remain elusive. We investigated IL6, C-reactive protein, and circulating cell-free mtDNA in serum of 245 participants in two cohorts from tertiary movement disorder centres. We performed a hypothesis-driven rank-based statistical approach adjusting for multiple testing. We detected (i) elevated IL6 levels in patients with biallelic PRKN/PINK1 mutations compared to healthy control subjects in a German cohort, supporting the concept of a role for inflammation in PRKN/PINK1-linked Parkinson's disease. In addition, the comparison of patients with biallelic and heterozygous mutations in PRKN/PINK1 suggests a gene dosage effect. The differences in IL6 levels were validated in a second independent Italian cohort; (ii) a correlation between IL6 levels and disease duration in carriers of PRKN/PINK1 mutations, while no such association was observed for idiopathic Parkinson's disease patients. These results highlight the potential of IL6 as progression marker in Parkinson's disease due to PRKN/PINK1 mutations; (iii) increased circulating cell-free mtDNA serum levels in both patients with biallelic or with heterozygous PRKN/PINK1 mutations compared to idiopathic Parkinson's disease, which is in line with previous findings in murine models. By contrast, circulating cell-free mtDNA concentrations in unaffected heterozygous carriers of PRKN/PINK1 mutations were comparable to control levels; and (iv) that circulating cell-free mtDNA levels have good predictive potential to discriminate between idiopathic Parkinson's disease and Parkinson's disease linked to heterozygous PRKN/PINK1 mutations, providing functional evidence for a role of heterozygous mutations in PRKN or PINK1 as Parkinson's disease risk factor. Taken together, our study further implicates inflammation due to impaired mitophagy and subsequent mtDNA release in the pathogenesis of PRKN/PINK1-linked Parkinson's disease. In individuals carrying mutations in PRKN/PINK1, IL6 and circulating cell-free mtDNA levels may serve as markers of Parkinson's disease state and progression, respectively. Finally, our study suggests that targeting the immune system with anti-inflammatory medication holds the potential to influence the disease course of Parkinson's disease, at least in this subset of patients.


Soluble antigen abrogates the appearance of anti-protein IgG1-forming cell precursors during primary immunization.
The anti-human serum albumin (HSA) B-cell repertoire of C57BL/6 mice was examined by culturing splenocytes at limiting dilution following polyclonal stimulation with Escherichia coli lipopolysaccharide and a lymphokine mixture. The frequency of anti-HSA precursors was determined before and after immunization with alum-precipitated HSA and 10(9) killed Bordetella pertussis organisms, by submitting clonal supernatants to an ELISA. Anti-HSA IgG1-forming precursors were rare in unimmunized spleens, representing approximately equal to 1 in 500,000 splenocytes or only approximately equal to 100 cells per spleen. Between day 5 and day 7 after immunization, this figure increased to approximately equal to 20,000 cells per spleen. Over the following 3 weeks, there was a progressive increase in the mean optical density generated in the clonal ELISA, presumably due to affinity maturation of the B-cell population. When freshly deaggregated HSA was injected before or even up to 4 days after challenge immunization, the appearance of anti-HSA IgG1-forming cell precursors was largely prevented. The effect was most marked with 5 mg or 1 mg of soluble HSA, but impressive partial effects could be seen with as little as 10 micrograms of HSA if administered before challenge immunization. Most of the few clones seen after the higher doses of the toleragen appeared to make antibody of low affinity. The capacity to influence the B-cell pool by soluble antigen administered just 1-2 days before the sudden appearance of IgG1 precursors argues against the totality of the effect being due to T-cell-mediated suppression and in favor of a direct effect on B cells.


Photoelectrochemical Behavior of a Molecular Ru-Based Water-Oxidation Catalyst Bound to TiO2-Protected Si Photoanodes.
A hybrid photoanode based on a molecular water oxidation precatalyst was prepared from TiO2-protected n- or p+-Si coated with multiwalled carbon nanotubes (CNT) and the ruthenium-based water oxidation precatalyst [RuIV(tda)(py-pyr)2(O)], 1(O) (tda2- is [2,2':6',2″-terpyridine]-6,6″-dicarboxylato and py-pir is 4-(pyren-1-yl)-N-(pyridin-4-ylmethyl)butanamide). The Ru complex was immobilized by π-π stacking onto CNTs that had been deposited by drop casting onto Si electrodes coated with 60 nm of amorphous TiO2 and 20 nm of a layer of sputtered C. At pH = 7 with 3 Sun illumination, the n-Si/TiO2/C/CNT/[1+1(O)] electrodes exhibited current densities of 1 mA cm-2 at 1.07 V vs NHE. The current density was maintained for >200 min at a constant potential while intermittently collecting voltammograms that indicated that over half of the Ru was still in molecular form after O2 evolution.


Single-shot interferometric measurement of pulse-to-pulse stability of absolute phase using a time-stretch technique.
Measurement of the absolute phase of ultrashort optical pulses in real-time is crucial for various applications, including frequency comb and high-field physics. Modern single-shot techniques, such as dispersive Fourier transform and time-lens, make it possible to investigate non-repetitive spectral dynamics of ultrashort pulses yet do not provide the information on absolute phase. In this work, we demonstrate a novel approach to characterise single-shot pulse-to-pulse stability of the absolute phase with the acquisition rate of 15 MHz. The acquisition rate, limited by the repetition rate of the used free-running mode-locked Erbium-doped fibre laser, substantially exceeds one of the traditional techniques. The method is based on the time-stretch technique. It exploits a simple all-fibre Mach-Zehnder interferometric setup with a remarkable resolution of ∼7.3 mrad. Using the proposed method, we observed phase oscillations in the output pulses governed by fluctuations in the pulse intensity due to Kerr-induced self-phase modulation at frequencies peaked at 4.6 kHz. As a proof-of-concept application of the demonstrated interferometric methodology, we evaluated phase behaviour during vibration exposure on the laser platform. The results propose a new view on the phase measurements that provide a novel avenue for numerous sensing applications with MHz data frequencies.


Patients' experiences of acquiring a deep surgical site infection: an interview study.
The negative impact of surgical site infection (SSI) in terms of morbidity, mortality, additional costs, and length of stay (LOS) in the hospital is well described in the literature, as are risk factors and preventive measures. Given the lack of knowledge regarding patients' experiences of SSI, the aim of the present study was to describe patients' experiences of acquiring a deep SSI. Content analysis was used to analyze data obtained from 14 open interviews with participants diagnosed with a deep SSI. Patients acquiring a deep SSI suffer significantly from pain, isolation, and insecurity. The SSI changes physical, emotional, social, and economic aspects of life in extremely negative ways, and these changes are often persistent. Health care professionals should focus on strategies to enable early diagnosis and treatment of SSIs. The unacceptable suffering related to the infection, medical treatment, and an insufficient patient-professional relationship should be addressed when planning individual care, because every effort is needed to support this group of patients and minimize their distress. All possible measures should be taken to avoid bacterial contamination of the surgical wound during and after surgery to prevent the development of SSI.


Roughing filtration as an effective pre-treatment system for high turbidity water.
Effective water treatment is the prime goal of every water treatment facility. Chakwal Water Treatment Plant in Pakistan has been treating high-turbidity surface water through crude coagulation, sedimentation and slow sand filtration since the early 1980s. The process has always been tedious in terms of high coagulant dosage, large volumes of sludge and short filter runs especially after wet spells. A laboratory-scale study was conducted to see if roughing filtration, as the pre-treatment process, would help in reducing coagulant dose and sludge volume and improving effluent quality. Results indicated that up-flow rouging filtration with media grades decreasing in the direction of flow could reduce wet weather raw water turbidity (by more than 90%) and coagulant dose. Overall, the plant could save over US $54,000 annually in terms of coagulant cost only. Longer filter runs, improved product water quality leading to lower chlorine dose requirement, would be additional benefits.


Safety and efficacy of iron therapy on reducing red blood cell transfusion requirements and treating anaemia in critically ill adults: A systematic review with meta-analysis and trial sequential analysis.
To evaluate the safety (risk of infection) and efficacy (transfusion requirements, changes in haemoglobin (Hb)) of iron therapy in adult intensive care unit (ICU) patients. We systematically searched seven databases for all relevant studies until January 2018 and included randomized (RCT) studies comparing iron, by any route, with placebo/no iron. 805 participants from 6 RCTs were included. Iron therapy, by any route, did not decrease the risk of requirement for a red blood cell (RBC) transfusion (Risk ratio (RR) 0.91, 95% CI 0.80 to 1.04, p = 0.15) or mean number of RBCs transfused per participant (mean difference (MD) -0.30, 95% CI -0.68 to 0.07, p = 0.15). Iron therapy did increase mean Hb concentration (MD 0.31 g/dL, 95% CI 0.04 to 0.59, p = 0.03). There was no difference in infection (RR 0.95, 95% CI 0.79 to 1.19, p = 0.44). Trial Sequential Analysis suggests that the required participant numbers to detect or reject a clinically important effect of iron therapy on transfusion requirements or infection in ICU patients has not yet been reached. Iron therapy results in a modest increase in Hb. The current evidence is inadequate to exclude an important effect on transfusion requirements or infection.


Evaluation of membrane fouling mitigation strategies in an algal membrane photobioreactor (AMPBR) treating secondary wastewater effluent.
Microalgae-based advanced wastewater treatment has gained momentum due to the possibility of recovering nutrients for the production of fertilizers, biofuels and fine chemicals from microalgal biomass. The objective of this study is to evaluate the effect of different fouling control strategies on the development of Chlorella vulgaris in a membrane photobioreactor (AMPMBR) treating a secondary wastewater effluent. The experimental results showed a decrease in the fouling rate (bar/hours) of 50% for backwash and relaxation and 60% for nitrogen bubble scouring. Additionally, in-situ non-destructive real time monitoring was employed to visualize and assess the change in morphology of the algae formed on the membrane surface. The use of fouling mitigation led to substantial changes in the biomass morphologies impacting the performance of the AMPMBR. The lowest biomass deposition (5-10 µm) was observed when nitrogen bubble scouring was employed, while the application of relaxation led to the thickest (180 µm), most heterogeneous and porous structure. The use of backwash led to a partial temporary biomass detachment from the membrane surface. This study, provided a better understanding of the impact of fouling mitigation strategies on the biomass formed on the membrane of AMPMBR.


Knockout of the rodent malaria parasite chitinase pbCHT1 reduces infectivity to mosquitoes.
During mosquito transmission, malaria ookinetes must cross a chitin-containing structure known as the peritrophic matrix (PM), which surrounds the infected blood meal in the mosquito midgut. In turn, ookinetes produce multiple chitinase activities presumably aimed at disrupting this physical barrier to allow ookinete invasion of the midgut epithelium. Plasmodium chitinase activities are demonstrated targets for human and avian malaria transmission blockade with the chitinase inhibitor allosamidin. Here, we identify and characterize the first chitinase gene of a rodent malaria parasite, Plasmodium berghei. We show that the gene, named PbCHT1, is a structural ortholog of PgCHT1 of the avian malaria parasite Plasmodium gallinaceum and a paralog of PfCHT1 of the human malaria parasite Plasmodium falciparum. Targeted disruption of PbCHT1 reduced parasite infectivity in Anopheles stephensi mosquitoes by up to 90%. Reductions in infectivity were also observed in ookinete feeds-an artificial situation where midgut invasion occurs before PM formation-suggesting that PbCHT1 plays a role other than PM disruption. PbCHT1 null mutants had no residual ookinete-derived chitinase activity in vitro, suggesting that P. berghei ookinetes express only one chitinase gene. Moreover, PbCHT1 activity appeared insensitive to allosamidin inhibition, an observation that raises questions about the use of allosamidin and components like it as potential malaria transmission-blocking drugs. Taken together, these findings suggest a fundamental divergence among rodent, avian, and human malaria parasite chitinases, with implications for the evolution of Plasmodium-mosquito interactions.


Preliminary analysis of the genetic basis for vancomycin resistance in Staphylococcus aureus strain Mu50.
Glycopeptides, such as vancomycin, are frequently the antibiotics of choice for treatment of infections caused by the now common methicillin-resistant Staphylococcus aureus (MRSA). Incidences of vancomycin resistance in S. aureus (VRSA) have been increasing worldwide for the last 5 years. Complex mechanisms producing changes in cell wall content and composition generate the VRSA phenotype, but the genetic basis of these changes has not yet been determined. To facilitate the genetic investigation, entire genome sequences of the archetypal VRSA (Mu50), and vancomycin-susceptible MRSA strains N315, EMRSA 16 and COL were compared. The in silico analysis revealed several loss-of-function mutations in Mu50, affecting important cell wall biosynthesis and intermediary metabolism genes, not previously reported. The new findings provide further evidence for the hypothesis that vancomycin resistance in Mu50 is due to fundamental changes, important to metabolic pathways that impinge on peptidoglycan biosynthesis. These observations will inform targeted experiments aimed at a complete understanding of the mechanism(s) of vancomycin resistance in S. aureus Mu50 and other VRSA strains.


Reading trajectories of children with language difficulties from preschool through fifth grade.
The current work examined which theory of reading development, the cumulative reading trajectory or the compensatory trajectory of development, most accurately represents the reading trajectories of children with language difficulties (LD) relative to their peers with typical language (TL) skills. Specifically, initial levels of reading skills, overall rate of growth, and patterns of growth were examined. Children were classified according to whether or not they exhibited LD at 54 months of age (LD n = 145; TL n = 653), using data from the National Institute of Child Health and Human Development's Early Child Care Research Network (see NICHD, 1993). A latent shape growth curve model was used to model reading skills at 4 time points from preschool through fifth grade. In comparison to children with TL, children with LD showed lower reading skills in preschool, but their overall reading growth was faster. All children developed the skills associated with reading more rapidly at earlier ages compared to later ages. Children with LD continued to exhibit reading skills that were substantially lower than those of children with TL during fifth grade. Results supported the compensatory trajectory of development. Speech-language pathologists are encouraged to adopt evidence-based practices in order to boost reading outcomes for children with LD beginning in preschool.


Epidemiology of lobomycosis-like disease in bottlenose dolphins Tursiops spp. from South America and southern Africa.
We report on the epidemiology of lobomycosis-like disease (LLD), a cutaneous disorder evoking lobomycosis, in 658 common bottlenose dolphins Tursiops truncatus from South America and 94 Indo-Pacific bottlenose dolphins T. aduncus from southern Africa. Photographs and stranding records of 387 inshore residents, 60 inshore non-residents and 305 specimens of undetermined origin (inshore and offshore) were examined for the presence of LLD lesions from 2004 to 2015. Seventeen residents, 3 non-residents and 1 inshore dolphin of unknown residence status were positive. LLD lesions appeared as single or multiple, light grey to whitish nodules and plaques that may ulcerate and increase in size over time. Among resident dolphins, prevalence varied significantly among 4 communities, being low in Posorja (2.35%, n = 85), Ecuador, and high in Salinas, Ecuador (16.7%, n = 18), and Laguna, Brazil (14.3%, n = 42). LLD prevalence increased in 36 T. truncatus from Laguna from 5.6% in 2007-2009 to 13.9% in 2013-2014, albeit not significantly. The disease has persisted for years in dolphins from Mayotte, Laguna, Salinas, the Sanquianga National Park and Bahía Málaga (Colombia) but vanished from the Tramandaí Estuary and the Mampituba River (Brazil). The geographical range of LLD has expanded in Brazil, South Africa and Ecuador, in areas that have been regularly surveyed for 10 to 35 yr. Two of the 21 LLD-affected dolphins were found dead with extensive lesions in southern Brazil, and 2 others disappeared, and presumably died, in Ecuador. These observations stress the need for targeted epidemiological, histological and molecular studies of LLD in dolphins, especially in the Southern Hemisphere.


Evaluation of the effect of diode laser for debonding ceramic brackets on nanomechanical properties of enamel.
The heat from laser can lead to the destruction of organic components of enamel and consequently changes in the mechanical properties of enamel. The purpose of this study was to evaluate the effect of diode laser on the nanomechanical properties of enamel in the process of debonding the ceramic brackets. In This in vitro study Eighteen ceramic brackets were bonded on the intact premolars in 3 groups of 6 (one control and two study groups). To debond the brackets in the study groups, laser diode was used for 3 s with 1W and 3W power. Shear bond strength and adhesive remnant index were recorded for all groups. Hardness and elastic modulus were measured in 1-31 μ depth from enamel surface in each debonded area. Analysis of variance was used for determining the difference in shear bond strength (SBS), hardness, and elastic modulus and was followed by post hoc Tukey's honest significant difference test. One-sample t-test was used to compare the changes in the pulp temperature with the standard threshold (5.5°C). The significance level was set at %5 in this study. SBS was significantly greater in the control group compared to the study groups. There was no significant difference in the average of hardness and elastic modulus of enamel between the groups. Pulp temperature elevation in the study groups was significantly < 5.5°C (P = 0.000). The diode laser with either 1W or 3W power for 3 s is effective in debonding the ceramic brackets without any detrimental effect on the pulp or mechanical properties of enamel. In regard to the pulp health, the 1W power laser is rather recommended for debonding the ceramic brackets than the 3W laser power.


Twelve-year trends in the prevalence and risk factors of diabetes and prediabetes in Turkish adults.
There is concern about an emerging diabetes epidemic in Turkey. We aimed to determine the prevalence of diagnosed and undiagnosed diabetes, prediabetes and their 12-year trends and to identify risk factors for diabetes in the adult Turkish population. A cross-sectional, population-based survey, 'TURDEP-II' included 26,499 randomly sampled adults aged ≥ 20 years (response rate: 87 %). Fasting glucose and biochemical parameters were measured in all; then a OGTT was performed to identify diabetes and prediabetes in eligible participants. The prevalence of diabetes was 16.5 % (new 7.5 %), translating to 6.5 million adults with diabetes in Turkey. It was higher in women than men (p = 0.008). The age-standardized prevalence to the TURDEP-I population (performed in 1997-98) was 13.7 % (if same diagnostic definition was applied diabetes prevalence is calculated 11.4 %). The prevalence of isolated-IFG and impaired glucose tolerance (IGT), and combined prediabetes was 14.7, 7.9, and 8.2 %, respectively; and that of obesity 36 % and hypertension 31.4 %. Compared to TURDEP-I; the rate of increase for diabetes: 90 %, IGT: 106 %, obesity: 40 % and central obesity: 35 %, but hypertension decreased by 11 % during the last 12 years. In women age, waist, body mass index (BMI), hypertension, low education, and living environment; in men age, BMI, and hypertension were independently associated with an increased prevalence of diabetes. In women current smoking, and in men being single were associated with a reduced risk. These results from one of the largest nationally representative surveys carried out so far show that diabetes has rapidly become a major public health challenge in Turkey. The figures are alarming and underscore the urgent need for national programs to prevent diabetes, to manage the illness and thus prevent complications.


Whooping cough and pertussis vaccine: a comparison of risks and benefits in Britain during the period 1968-83.
Since 1975, acceptance of pertussis vaccine has fallen from over 70% to 50% or less in most parts of Britain. This permits evaluation of a continuing natural experiment in which the frequency and severity of whooping cough can be compared those of adverse events following injections of pertussis vaccine. National data show an increase in notifications of whooping cough in most parts of Britain since 1975. Hospital admissions show considerable variation between areas with relatively high rates in some areas of deprivation but very low rates in more affluent areas even where vaccine-acceptance is around 50%. Deaths of infants with whooping cough have decreased steadily since 1900, the rate since 1975 being the lowest ever. Active epidemiological surveillance in Glasgow, with a population of 216,000 children and 13,000 births annually, shows that outbreaks and severe cases requiring admission to hospital were concentrated consistently in a few areas of deprivation. There is a significant correlation between vaccine-acceptance and hospital admission by district of residence but analysis of variance shows this effect to be less than that of overcrowding in households and other deprivation variables. In each of three outbreaks studied prospectively (1974-5, 78-78 and 82) about 30% of cases occurred in children who had received three doses of pertussis vaccine. Such vaccination had a significant protective effect in children aged 1-4 years but not in older children. There was no evidence of a herd immunity sufficient to protect infants below age for vaccination. Admissions to hospital decreased during the period 1970-83. There were no deaths attributable to proven or suspected infections with Bordetella pertussis during the period 1972-1983. No cases of encephalopathy, permanent brain damage or lung damage were detected in a follow up of all cases notified, surveyed or admitted to hospital between 1975 and 1982. Collectively, these national and local data provided estimates of the frequency of infection, complications of infection, admission to hospital and death in children with whooping cough for comparison with local, national and published estimates of the frequency and severity of adverse reactions, encephalopathy, permanent brain damage and death after injections of pertussis vaccine. It is concluded that, in children living in non-deprived circumstances in Britain, the risk of pertussis vaccine during the period 1970-83 exceeded those of whooping cough. In some deprived sectors, the risks from whooping cough might have been marginally higher but there was no evidence that this was associated with any increase in deaths or permanent disabilities.


How birds dissipate heat before, during and after flight.
Animal flight uses metabolic energy at a higher rate than any other mode of locomotion. A relatively small proportion of the metabolic energy is converted into mechanical power; the remainder is given off as heat. Effective heat dissipation is necessary to avoid hyperthermia. In this study, we measured surface temperatures in lovebirds (Agapornis personatus) using infrared thermography and used heat transfer modelling to calculate heat dissipation by convection, radiation and conduction, before, during and after flight. The total non-evaporative rate of heat dissipation in flying birds was 12× higher than before flight and 19× higher than after flight. During flight, heat was largely dissipated by forced convection, via the exposed ventral wing areas, resulting in lower surface temperatures compared with birds at rest. When perched, both before and after exercise, the head and trunk were the main areas involved in dissipating heat. The surface temperature of the legs increased with flight duration and remained high on landing, suggesting that there was an increase in the flow of warmer blood to this region during and after flight. The methodology developed in this study to investigate how birds thermoregulate during flight could be used in future studies to assess the impact of climate change on the behavioural ecology of birds, particularly those species undertaking migratory flights.


An innocuous bias in whisker use in adult rats modifies receptive fields of barrel cortex neurons.
The effect of innocuously biasing the flow of sensory activity from the whiskers for periods of 3-30 d in awake, behaving adult rats on the receptive field organization of rat SI barrel cortex neurons was studied. One pair of adjacent whiskers, D2 and either D1 or D3, remained intact unilaterally (whisker pairing), all others being trimmed throughout the period of altered sensation. Receptive fields of single cells in the contralateral D2 barrel were analyzed under urethane anesthesia by peristimulus time histogram (PSTH) and latency histogram analysis after 3, 7-10, and 30 d of pairing and compared with controls, testing all whiskers cut to the same length. Response magnitudes to surround receptive field in-row whiskers D1 and D3 were not significantly different for control animals. The same was found for surround in-arc whiskers C2 and E2. However, after 3 d of whisker pairing a profound bias occurred in response to the paired D-row surround whisker relative to the opposite trimmed surround D-row whisker and to the C2 and E2 whiskers. This bias increased with the duration of pairing, regardless of which surround whisker (D1 or D3) was paired with D2. For all three periods of pairing the mean response to the paired surround whisker was increased relative to controls, but peaked at 7-10 d. Response to the principal center-receptive (D2) whisker was increased for the 3 and 7-10 d groups and then decreased at 30 d. Responses to trimmed arc surround whiskers (C2 and E2) were decreased in proportion to the duration of changed experience. Analysis of PSTH data showed that earliest discharges (5-10 msec poststimulus) to the D2 whisker increased progressively in magnitude with duration of pairing. For the paired surround whisker similar early discharges newly appeared after 30 d of pairing. At 3 and 7-10 d of pairing, increases in response to paired whiskers and decreases to cut surround whiskers were confined to late portions of the PSTH (10-100 msec poststimulus). Changes at 3-10 d can be attributed to alterations in intracortical synaptic relay between barrels. Longer-term changes in response to both paired whisker inputs (30 d) largely appear to reflect increases in thalamocortical synaptic efficacy. Our findings suggest that novel innocuous somatosensory experiences produce changes in the receptive field configuration of cortical cells that are consistent with Hebbian theories of experience-dependent potentiation and weakening of synaptic efficacy within SI neocortical circuitry, for correlated and uncorrelated sensory inputs, respectively.


Palliative Care with Attachment Hybrid Removable Prosthesis.
Abutment injury, unsatisfactory aesthetics and lesser retention exist with the cast partial denture. Though these constraints exist in the Removable Partial Denture (RPD) it is still widely used because of the simplicity in design, fabrication, economics and patient comfort. This clinical report describes a hybrid RPD technique which uses extra coronal attachment that reduces the limitations and provides better comfort for the patient.


Association between Daily Hydrogen Sulfide Exposure and Incidence of Emergency Hospital Visits: A Population-Based Study.
The adverse health effects of high concentrations of hydrogen sulfide (H2S) exposure are well known, though the possible effects of low concentrations have not been thoroughly studied. The aim was to study short-term associations between modelled ambient low-level concentrations of intermittent hydrogen sulfide (H2S) and emergency hospital visits with heart diseases (HD), respiratory diseases, and stroke as primary diagnosis. The study is population-based, using data from patient-, and population-registers from the only acute care institution in the Reykjavik capital area, between 1 January, 2007 and 30 June, 2014. The study population was individuals (≥18yr) living in the Reykjavik capital area. The H2S emission originates from a geothermal power plant in the vicinity. A model was used to estimate H2S exposure in different sections of the area. A generalized linear model assuming Poisson distribution was used to investigate the association between emergency hospital visits and H2S exposure. Distributed lag models were adjusted for seasonality, gender, age, traffic zones, and other relevant factors. Lag days from 0 to 4 were considered. The total number of emergency hospital visits was 32961 with a mean age of 70 years. In fully adjusted un-stratified models, H2S concentrations exceeding 7.00μg/m3 were associated with increases in emergency hospital visits with HD as primary diagnosis at lag 0 risk ratio (RR): 1.067; 95% confidence interval (CI): 1.024-1.111, lag 2 RR: 1.049; 95%CI: 1.005-1.095, and lag 4 RR: 1.046; 95%CI: 1.004-1.089. Among males an association was found between H2S concentrations exceeding 7.00μg/m3, and HD at lag 0 RR: 1.087; 95%CI: 1.032-1.146 and lag 4 RR: 1080; 95%CI: 1.025-1.138; and among those 73 years and older at lag 0 RR: 1.075; 95%CI: 1.014-1.140 and lag 3 RR: 1.072; 95%CI: 1.009-1.139. No associations were found with other diseases. The study showed an association between emergency hospital visits with HD as primary diagnosis and same day H2S concentrations exceeding 7.00μg/m3, more pronounced among males and those 73 years and older than among females and younger individuals.


Single-leg-stance radiographs in the diagnosis of pelvic instability.
In the nonacute setting, the diagnosis of pelvic instability is difficult. Patients who present with pelvic pain may have underlying instability. The purpose of the present study was to report the effectiveness of single-leg-stance radiographs in the diagnosis of pelvic instability in a consecutive series of patients presenting with pelvic pain. Thirty-eight consecutive patients (twenty-four women and fourteen men) ranging in age from eighteen to seventy-eight years who presented with pelvic pain and a history of injury (twenty-seven), childbirth (seven [four primiparous and three multiparous]), or osteopenia (four) were evaluated with a visual analog scale pain score and a standard series of radiographs in an attempt to identify pelvic instability. The average time from the onset of symptoms to the evaluation was forty-one months (range, six weeks to twenty-seven years). Each patient was evaluated with supine anteroposterior, inlet, and outlet pelvic radiographs; a standing anteroposterior pelvic radiograph; and two single-leg-standing pelvic radiographs (one with the patient standing on the left leg and one with the patient standing on the right leg). A positive finding was defined as >or=0.5 cm of vertical translation measured at the symphyseal bodies between the two single-leg-stance radiographs. Of the thirty-eight patients, twenty-five demonstrated pelvic instability (average, 1.98 cm; range, 0.5 to 5 cm). With the numbers available, the average visual analog scale pain score for the patients with a stable pelvis was not significantly different from that for the patients with an unstable pelvis (6.4 +/- 2.9 compared with 7.3 +/- 1.9; p = 0.28). Standing anteroposterior and single-leg-stance pelvic radiographs aid in the diagnosis of pelvic instability more effectively than do the standard three radiographs of the pelvis made in the supine position or a standing anteroposterior radiograph of the pelvis alone. Additional studies will be needed to correlate this instability with clinical symptoms. Diagnostic Level IV. See Instructions to Authors for a complete description of levels of evidence.


Establishing usability heuristics for heuristics evaluation in a specific domain: Is there a consensus?
Heuristics evaluation is frequently employed to evaluate usability. While general heuristics are suitable to evaluate most user interfaces, there is still a need to establish heuristics for specific domains to ensure that their specific usability issues are identified. This paper presents a comprehensive review of 70 studies related to usability heuristics for specific domains. The aim of this paper is to review the processes that were applied to establish heuristics in specific domains and identify gaps in order to provide recommendations for future research and area of improvements. The most urgent issue found is the deficiency of validation effort following heuristics proposition and the lack of robustness and rigour of validation method adopted. Whether domain specific heuristics perform better or worse than general ones is inconclusive due to lack of validation quality and clarity on how to assess the effectiveness of heuristics for specific domains. The lack of validation quality also affects effort in improving existing heuristics for specific domain as their weaknesses are not addressed.


β-Lactoglobulin Is Insulinotropic Compared with Casein and Whey Protein Ingestion during Catabolic Conditions in Men in a Double-Blinded Randomized Crossover Trial.
Muscle loss during acute infectious disease is mainly triggered by inflammation, immobilization, and malnutrition. The objective was to compare muscle protein kinetics and metabolism following ingestion of the dairy protein supplements β-lactoglobulin (BLG), casein (CAS), and whey (WHE) during controlled catabolic conditions. We used a randomized crossover design (registered at clinicaltrials.gov as NCT03319550) to investigate 9 healthy male participants [age: 20-40 y; BMI (in kg/m2) 20-30] who were randomly assigned servings of BLG, CAS, or WHE (0.6 g protein/kg, one-third as bolus and two-thirds as sip every 20 min) on 3 separate occasions separated by ∼6-8 wk. The participants received an infusion of lipopolysaccharide (1 ng/kg) combined with 36 h of fasting and bed rest before each study day, mimicking a clinical catabolic condition. The forearm model and isotopic tracer techniques were used to quantify muscle protein kinetics. Muscle biopsy specimens were obtained and intramyocellular signaling investigated using Western blot. BLG, CAS, and WHE improved the net balance of phenylalanine (NBphe) from baseline with ∼75% (P < 0.001) with no difference between interventions (primary outcome, P < 0.05). No difference in rates of appearance and disappearance of phenylalanine or in intramyocellular signaling activation was found between interventions (secondary outcomes). The incremental AUC for serum insulin was 62% higher following BLG compared with CAS (P < 0.001) and 30% higher compared with WHE (P = 0.002), as well as 25% higher in WHE compared with CAS (P = 0.006). Following BLG consumption, plasma concentrations of glucose-dependent insulinotropic peptide (GIP) increased 70% compared with CAS (P = 0.001) and increased 34% compared with WHE (P = 0.06). No significant difference was found between WHE and CAS (P = 0.12). BLG, WHE, and CAS have similar effects on muscle in young male participants during catabolic conditions. BLG showed specific, possibly GIP-dependent, insulinotropic properties, which may have future clinical implications.


A paradigm for consensus. The University Hospital Consortium guidelines for the use of albumin, nonprotein colloid, and crystalloid solutions.
To develop contemporary, comprehensive guidelines for the appropriate and efficient use of albumin, nonprotein colloid, and crystalloid solutions. A systematic, literature-based, consensus exercise employing a modified Delphi method. Thirty-one medical and allied health professionals from 26 University Hospital Consortium (Oak Brook, Ill) member institutions were initially chosen to participate. Participants were selected on the basis of their recognized research in the use of albumin, nonprotein colloid, and crystalloid solutions, and/or experience in the review of appropriateness of such use. A total of 24 participants completed the exercise. Group responses were statistically analyzed in an iterative consensus development process. Five separate questionnaire rounds were designed to establish criteria for the appropriate use of albumin, nonprotein colloid, and crystalloid solutions. Consensus guidelines were developed outlining the appropriate use of these products for 12 clinical indications, including hemorrhagic shock, nonhemorrhagic (maldistributive) shock, hepatic resection, thermal injury, cerebral ischemia, nutritional intervention, cardiac surgery, hyperbilirubinemia of the newborn, cirrhosis and paracentesis, nephrotic syndrome, organ transplantation, and plasmapheresis. The Delphi method, a systematic, literature-based consensus process, was shown to be useful in the development of complex clinical practice guidelines for the use of albumin, nonprotein colloid, and crystalloid solutions. It is anticipated that the guidelines will assist health care providers to develop local institutional policies and procedures for the appropriate and efficient use of albumin and albumin alternatives. Institutions reviewing and updating existing local guidelines may use the University Hospital Consortium guidelines as a model for comparison.


Measurement and clinical monitoring of human lymphocyte clonality by massively parallel VDJ pyrosequencing.
The complex repertoire of immune receptors generated by B and T cells enables recognition of diverse threats to the host organism. In this work, we show that massively parallel DNA sequencing of rearranged immune receptor loci can provide direct detection and tracking of immune diversity and expanded clonal lymphocyte populations in physiological and pathological contexts. DNA was isolated from blood and tissue samples, a series of redundant primers was used to amplify diverse DNA rearrangements, and the resulting mixtures of barcoded amplicons were sequenced using long-read ultra deep sequencing. Individual DNA molecules were then characterized on the basis of DNA segments that had been joined to make a functional (or nonfunctional) immune effector. Current experimental designs can accommodate up to 150 samples in a single sequence run, with the depth of sequencing sufficient to identify stable and dynamic aspects of the immune repertoire in both normal and diseased circumstances. These data provide a high-resolution picture of immune spectra in normal individuals and in patients with hematological malignancies, illuminating, in the latter case, both the initial behavior of clonal tumor populations and the later suppression or re-emergence of such populations after treatment.


Improved sensitivity of vaginal self-collection and high-risk human papillomavirus testing.
Self-collected vaginal specimens tested for high-risk human papillomavirus (HR-HPV) have been shown to be less sensitive for the detection of cervical intraepithelial neoplasia or cancer (≥CIN 3) than physician-collected endocervical specimens. To increase the sensitivity of self-collected specimens, we studied a self-sampling device designed to obtain a larger specimen from the upper vagina (POI/NIH self-sampler) and a more sensitive polymerase chain reaction (PCR)-based HR-HPV assay. Women (10,000) were screened with cervical cytology and HR-HPV testing of vaginal self-collected and endocervical physician-collected specimens. Women were randomly assigned to use either a novel self-collection device (POI/NIH self-sampler) or conical-shaped brush (Qiagen). The self-collected and clinician-collected specimens were assayed by Cervista (Hologic) and the research only PCR-based matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF). Women with any abnormal screening test underwent colposcopy and biopsy. Women (8,556), mean age of 38.9, had complete data; 1.6% had ≥ CIN 3. For either HR-HPV assay, the sensitivity was similar for the two self-collection devices. Tested with Cervista, the sensitivity for ≥CIN 3 of self-collected specimens was 70.9% and for endocervical specimens was 95.0% (p = 0.0001). Tested with MALDI-TOF, the sensitivity for ≥CIN 3 of self-collected specimens was 94.3% and for endocervical specimens was also 94.3% (p = 1.0). A self-collected sample using a PCR-based assay with the capability of very high throughput has similar sensitivity as a direct endocervical specimen obtained by a physician. Large population-based screening "events" in low-resource settings could be achieved by promoting self-collection and centralized high-throughput, low-cost testing by PCR-based MALDI-TOF.


A large-depth-of-field projected fringe profilometry using supercontinuum light illumination.
In this paper, a large-depth-of-field projected fringe profilometry using a supercontinuum light source generated by launching femto second laser pulses into a highly nonlinear photonic crystal fiber is presented. Since the supercontinuum light has high spatial coherence and a broad spectral range (from UV to near infrared), a high power (hundreds of mW) point white light source can be employed to generate modulated fringe patterns, which offers following major advantages: (1) large-depth-of-field, (2) ease of calibration, and (3) little speckle noise (a major problem for the laser system). Thus, a highly accurate, large-depth-of-field projected fringe profilometer can be realized. Both the theoretical description and experimental demonstration are provided.


Relationship between red meat allergy and sensitization to gelatin and galactose-α-1,3-galactose.
We have observed patients clinically allergic to red meat and meat-derived gelatin. We describe a prospective evaluation of the clinical significance of gelatin sensitization, the predictive value of a positive test result, and an examination of the relationship between allergic reactions to red meat and sensitization to gelatin and galactose-α-1,3-galactose (α-Gal). Adult patients evaluated in the 1997-2011 period for suspected allergy/anaphylaxis to medication, insect venom, or food were skin tested with gelatin colloid. In vitro (ImmunoCAP) testing was undertaken where possible. Positive gelatin test results were observed in 40 of 1335 subjects: 30 of 40 patients with red meat allergy (12 also clinically allergic to gelatin), 2 of 2 patients with gelatin colloid-induced anaphylaxis, 4 of 172 patients with idiopathic anaphylaxis (all responded to intravenous gelatin challenge of 0.02-0.4 g), and 4 of 368 patients with drug allergy. Test results were negative in all patients with venom allergy (n = 241), nonmeat food allergy (n = 222), and miscellaneous disorders (n = 290). ImmunoCAP results were positive to α-Gal in 20 of 24 patients with meat allergy and in 20 of 22 patients with positive gelatin skin test results. The results of gelatin skin testing and anti-α-Gal IgE measurements were strongly correlated (r = 0.46, P < .01). α-Gal was detected in bovine gelatin colloids at concentrations of approximately 0.44 to 0.52 μg/g gelatin by means of inhibition RIA. Most patients allergic to red meat were sensitized to gelatin, and a subset was clinically allergic to both. The detection of α-Gal in gelatin and correlation between the results of α-Gal and gelatin testing raise the possibility that α-Gal IgE might be the target of reactivity to gelatin. The pathogenic relationship between tick bites and sensitization to red meat, α-Gal, and gelatin (with or without clinical reactivity) remains uncertain.


Challenges Facing First-Generation College Graduates in Medical School: A Qualitative Analysis.
First-generation (FG) medical students remain underrepresented in medicine despite ongoing national efforts to increase diversity; understanding the challenges faced by this student population is essential to building holistic policies, practices, and learning environments that promote professional actualization. Although FG students have been extensively studied in the undergraduate literature, there is little research investigating how FG students experience medical education or opportunities for educators to intervene. To explore challenges that FG students experience in undergraduate medical education and identify opportunities to improve foundational FG support. This qualitative study was conducted using an online platform with 37 FG students enrolled in 27 US medical schools. An interprofessional team of medical educators and trainees conducted semistructured interviews from November 2021 through April 2022. Participants were recruited using a medical student listserv. Data were analyzed from April to November 2022. After conducting a preliminary analysis using open coding, a codebook was created and used in a thematic analysis; the codebook used a combination of deductive and inductive coding. Among the 37 students recruited for this study, 21 (56.8%) were female; 23 (62.2%) were in the clinical phase of training; 1 (2.7%) was American Indian or Alaska Native, 7 (18.9%) were Hispanic, Latino, or of Spanish origin, 8 (21.6%) were non-Hispanic Asian or Asian American, 9 (24.3%) were non-Hispanic Black or African American, and 23 (32.4%) were non-Hispanic White; mean (SD) age was 27.3 (2.8) years. Participants described 4 major themes: (1) isolation and exclusion related to being a newcomer to medicine; (2) difficulty with access to basic resources (eg, food, rent, transportation) as well as educational (eg, books); (3) overall lack of faculty or institutional support to address these challenges; and (4) a sense of needing to rely on grit and resilience to survive. Although grit and resilience are desirable traits, results of this study suggest that FG medical students face increased adversity with inadequate institutional support, which forces them to excessively rely on grit and resilience as survival (rather than educational) strategies. By applying the holistic model often used in admissions to the postmatriculation educational process, targeted and flexible initiatives can be created for FG students so that all students, regardless of background, can achieve robust professional actualization.


First Report of Fusarium Wilt Caused by Fusarium oxysporum f. sp. canariensis on Canary Island Date Palm in Texas and South Carolina.
Canary Island date palm (Phoenix canariensis) is native to the Canary Islands and widely grown throughout the world as an ornamental. At a home site in Austin, TX in May 2008 and a commercial site near Charleston, SC in December 2009, declining Canary Island date palms were observed. Symptoms included individual leaves with chlorotic or necrotic leaflets on one side of the leaf blade (one-sided wilt or death) and a distinct reddish brown stripe along the petiole and rachis. Cross-sections through the petiole or rachis exhibited discoloration of internal tissue. Fusarium oxysporum was isolated from the internal petiole or rachis tissue of each palm sample onto one-quarter-strength potato dextrose agar (PDA). Typical macroconidia in pale orange sporodochia, microconidia in false heads on short monophialides, and chlamydospores were observed (2). Macroconidia were mostly 3-septate, slightly curved, and ranged from 3.8 to 4.2 × 42.9 to 46.5 μm. Microconidia were single cell, oval to reniform, and ranged from 2.5 to 2.9 × 7.2 to 7.8 μm. Single-spore isolates grown on full-strength PDA (12-h light and 26°C) produced abundant white-to-pale lavender mycelia with a purple pigment in the agar. One isolate from each location (PLM-385B from Texas and PLM-511A from South Carolina) was selected for pathogenicity tests and molecular characterization. The translation elongation factor 1-α gene (EF-1α) was amplified in each isolate by PCR using the ef1 and ef2 primers (1). Products were sequenced and queried for similarity against the NCBI database and the FUSARIUM-ID database ( http://isolate.fusariumdb.org/index.php ) (1) using the BLAST search tool. In both databases, both isolates matched F. oxysporum f. sp. canariensis strain NRRL 26035 (GenBank Accession No. AF008485; FD_01211) at 100% sequence similarity. Sequences for PLM-385B and PLM-511A have been deposited in the NCBI database (GenBank Accession Nos. HM591537 and HM591538, respectively). Pathogenicity of these two isolates was tested on three-leaf Canary Island date palm seedlings. There were five replicate palms per isolate and control treatment. All potting mix was shaken from the roots and three groups of five seedlings were placed in small buckets. Twenty-five milliliters of a 106 conidia ml-1 suspension was pipetted down among the leaf bases and the excess drained onto the roots. Control palms received sterile water. Seedlings were covered with plastic for 48 h and then transplanted into separate growing containers. Ten weeks after inoculation, initial symptoms of a leaf wilt (off-color and folded over) were observed on some of the inoculated palms. After 4 months, all palms inoculated with PLM-511A were dead and three of the five palms inoculated with PLM-385B were dead. The pathogen was reisolated from diseased palms. All five control palms remained healthy. While the symptomatic palm in Texas had been in the home site approximately 2 years, which implied the palm could have been already infected when transplanted, the palm in South Carolina had been planted in 1990. To our knowledge, this is the first report of Fusarium wilt of Canary Island date palm in Texas and South Carolina. Previously in the United States, the disease had only been noted in California, Florida, and Nevada. References: (1) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.


Neutrophil accumulation and NET release contribute to thrombosis in HIT.
Heparin-induced thrombocytopenia (HIT) is an immune-mediated thrombocytopenic disorder associated with a severe prothrombotic state. We investigated whether neutrophils and neutrophil extracellular traps (NETs) contribute to the development of thrombosis in HIT. Using an endothelialized microfluidic system and a murine passive immunization model, we show that HIT induction leads to increased neutrophil adherence to venous endothelium. In HIT mice, endothelial adherence is enhanced immediately downstream of nascent venous thrombi, after which neutrophils undergo retrograde migration via a CXCR2-dependent mechanism to accumulate into the thrombi. Using a microfluidic system, we found that PF4 binds to NETs, leading them to become compact and DNase resistant. PF4-NET complexes selectively bind HIT antibodies, which further protect them from nuclease digestion. In HIT mice, inhibition of NET formation through Padi4 gene disruption or DNase treatment limited venous thrombus size. PAD4 inactivation did affect arterial thrombi or severity of thrombocytopenia in HIT. Thus, neutrophil activation contributes to the development of venous thrombosis in HIT by enhancing neutrophil-endothelial adhesion and neutrophil clot infiltration, where incorporated PF4-NET-HIT antibody complexes lead to thrombosis propagation. Inhibition of neutrophil endothelial adhesion, prevention of neutrophil chemokine-dependent recruitment of neutrophils to thrombi, or suppression of NET release should be explored as strategies to prevent venous thrombosis in HIT.


Radiation therapy and concomitant paclitaxel/carboplatin chemotherapy for muscle invasive transitional cell carcinoma of the bladder: a well-tolerated combination.
This review evaluates tolerance and disease control for eight patients with muscle invasive bladder cancer treated with pelvic radiotherapy and concomitant paclitaxel/carboplatin chemotherapy. From October 1996 through February 1998, eight patients were treated with pelvic radiotherapy and concomitant paclitaxel/carboplatin chemotherapy. All received from 39.60-41.40 Gy to the pelvis followed by a boost to the initial site of disease. Final tumor doses ranged from 64.80-68.40 Gy. Most patients received paclitaxel at 150 mg/m2 and carboplatin at an area under the curve (AUC) of 7 at 3-week cycles during the radiation therapy. No patient required treatment interruption. With a median follow-up of 27 months, three patients remain free of local and distant disease at follow-up intervals of 24, 25, and 31 months. No surviving locally controlled patient demonstrated late urinary or gastrointestinal morbidity. All patients with a visibly complete transurethral resection of bladder tumor (TURBT) prior to radiotherapy achieved local disease control. For this group of patients, the absolute 2-year pelvic tumor control rate is 57%. The 2-year disease-specific survival is 43%. Paclitaxel/carboplatin chemotherapy can be delivered with continuous course pelvic radiation therapy without severe acute or apparent late toxicity. This combination also appears to be effective in achieving disease control in the urinary bladder, particularly in those patients who have undergone a thorough TURBT. The authors believe that it would be reasonable to investigate this combination in future bladder conservation protocols. The combination of paclitaxel and carboplatin with radiotherapy may be of particular value in elderly patients or those with renal impairment.


Semiquantitative analysis of the histopathological features of the neuropathic foot ulcer: effects of pressure relief.
This study was designed to evaluate the histopathology of neuropathic ulcers and whether pressure relief could change such histological patterns. We compared neuropathic plantar ulcers tissue excised from 10 diabetic patients (group A) with those taken from 10 patients with comparable lesions and glycemic control after 20 days in a total contact cast (group B). Tissue specimens were blindly examined by two independent pathologists for hyperkeratosis, fibrosis, cutaneous annexes, capillaries, inflammation, cellular debris, and granulating tissue. For each parameter, quantification was obtained according to an arbitrary score: 0, absent; 1, present in <33%; 2, present in 34-66%; and 3, present in >67% of the lesion. Patients in group B showed a marked reduction in ulcer size after 20 days of casting (P < 0.01). The histopathological features of the two groups markedly differed. Group A patients showed a predominance of inflammatory elements as well as matrix alterations, vessel disruptions, inflammation, and debris. Group B ulcers showed a shift toward a reparative pattern with prevalence of neoformed capillaries and fibroblasts. Semiquantitative analysis confirmed the prevalence of hyperkeratosis, fibrosis, inflammation, and cellular debris in group A patients (P < 0.05), whereas cutaneous annexes, capillaries, and granulating tissue were more prevalent in group B lesions (P < 0.01). These results indicate that pressure relief with a total contact cast is associated with changes in the histology of neuropathic foot ulcers, indicating reduction of inflammatory and reactive components and acceleration of reparative processes.


Protective effects of Petroselinum crispum (Mill) Nyman ex A. W. Hill leaf extract on D-galactose-induced oxidative stress in mouse brain.
With an aim to examine the effect of ethanolic extract of P. crispum (Parsley) leaves on the D-galactose-induced oxidative stress in the brain of mouse, the activities of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) involved in oxygen radical (OR)-detoxification and antiperoxidative defense were measured in conjunction with an index of lipid peroxidation in mitochondrial fraction of various regions of the mouse brain. A significant decrease in superoxide dismutase and glutathione peroxidase activity was observed in D-galactose-stressed mice, while catalase activity was increased. Treatment of D-galactose-stressed mice with the ethanolic extract of P. crispum showed protection against the induced oxidative stress in brain regions. Concentration of thiobarbituric acid-reactive product was greatly elevated in D-galactose stress-induced mice and was significantly reduced in the brain regions of these mice upon treatment with P. crispum. It is postulated that parsley shows a protective effect against mitochondrial oxidative damage in the mouse brain.


Progression and risk factors of pododermatitis in part-time group housed rabbit does in Switzerland.
In rabbits (Oryctolagus cuniculus L.), pododermatitis is a chronic multifactorial skin disease that appears mainly on the plantar surface of the hind legs. This presumably progressive disease can cause pain leading to poor welfare, yet the progression of this disease has not been thoroughly assessed on the level of individual animals. The aim of this longitudinal study thus was to investigate the possible risk factors and the progression of pododermatitis in group housed breeding does in Switzerland on litter and plastic slats. Three commercial rabbit farms with part-time group housing on litter and plastic slats were visited every four weeks throughout one year. During every visit, the same 201 adult female breeding rabbits (67 does per farm) were evaluated for the presence and severity of pododermatitis. Additionally, the does' age, parity, body weight, reproductive state, hybrid, claw length, cleanliness and moisture of the paws and the temperature and humidity inside the barns were recorded as potential risk factors. The risk factors were analysed through general linear models and additive Bayesian network (ABN) modelling using a directed acyclic graph (DAG) for visualising associations between potential risk factors. The progression of pododermatitis was analysed with a transition matrix. Relative humidity inside the barns, body weight, number of kindlings, age, and claw length were the most important risk factors, all being positively associated with pododermatitis. In contrast to expectations, the cleanliness of the left hind paw was negatively associated with the occurrence of pododermatitis, but the effect was small. In young does, the severity of pododermatitis quickly increased and in some rabbits proceeded to ulcerated spots. It was shown that 60.00%, 14.17% and 3.33% of ulcerated lesions recovered to a state without ulceration within 4, 8 or >12 weeks, respectively.


Treatment sequencing in metastatic colorectal cancer.
Metastatic colorectal cancer (mCRC) remains incurable in most cases, but survival has improved with advances in cytotoxic chemotherapy and targeted agents. However, the optimal use and sequencing of these agents across multiple lines of treatment is unclear. Here, we review current treatment approaches and optimal treatment sequencing across the first-, second- and third-line settings in mCRC, including biological aspects affecting sequencing and rechallenge. Effective first-line therapy is a key determinant of treatment outcomes and should be selected after considering both clinical factors and biological markers, notably RAS and BRAF. The second-line regimen choice depends on the systemic therapies given in first-line. Anti-angiogenic agents (e.g. bevacizumab, ramucirumab and aflibercept) are indicated for most patients, whereas epidermal growth factor receptor (EGFR) inhibitors do not improve survival in the second-line setting. Molecular profiling is important in third-line treatment, with options in RAS wild-type patients including EGFR inhibitors (cetuximab or panitumumab), regorafenib and trifluridine/tipiracil. Immunotherapy with pembrolizumab or nivolumab ± ipilimumab may be considered for patients with high microsatellite instability disease. Targeting HER2/neu amplification shows promise for the subset of CRC tumours displaying this abnormality. Sequencing decisions are complicated by the potential for any treatment break or de-escalation to evoke a distinct clinical progression type. Ongoing trials are investigating the optimal sequencing and timing of therapies for mCRC. Molecular profiling has established new targets, and increasing knowledge of tumour evolution under drug pressure will possibly impact on sequencing.


A proteomic analysis of Pakistan Daboia russelii russelii venom and assessment of potency of Indian polyvalent and monovalent antivenom.
To address the dearth of knowledge on the biochemical composition of Pakistan Russell's Viper (Daboia russelii russelii) venom (RVV), the venom proteome has been analyzed and several biochemical and pharmacological properties of the venom were investigated. SDS-PAGE (reduced) analysis indicated that proteins/peptides in the molecular mass range of ~56.0-105.0kDa, 31.6-51.0kDa, 15.6-30.0kDa, 9.0-14.2kDa and 5.6-7.2kDa contribute approximately 9.8%, 12.1%, 13.4%, 34.1% and 30.5%, respectively of Pakistan RVV. Proteomics analysis of gel-filtration peaks of RVV resulted in identification of 75 proteins/peptides which belong to 14 distinct snake venom protein families. Phospholipases A2 (32.8%), Kunitz type serine protease inhibitors (28.4%), and snake venom metalloproteases (21.8%) comprised the majority of Pakistan RVV proteins, while 11 additional families accounted for 6.5-0.2%. Occurrence of aminotransferase, endo-β-glycosidase, and disintegrins is reported for the first time in RVV. Several of RVV proteins/peptides share significant sequence homology across Viperidae subfamilies. Pakistan RVV was well recognized by both the polyvalent (PAV) and monovalent (MAV) antivenom manufactured in India; nonetheless, immunological cross-reactivity determined by ELISA and neutralization of pro-coagulant/anticoagulant activity of RVV and its fractions by MAV surpassed that of PAV. The study establishes the proteome profile of the Pakistan RVV, thereby indicating the presence of diverse proteins and peptides that play a significant role in the pathophysiology of RVV bite. Further, the proteomic findings will contribute to understand the variation in venom composition owing to different geographical location and identification of pharmacologically important proteins in Pakistan RVV.


The economic burden of coronary heart disease in the UK.
To estimate the economic burden of coronary heart disease in the UK using both direct and indirect costs. A prevalence based approach was used to assess coronary heart disease related costs from the societal perspective. All UK residents in 1999 with coronary heart disease (ICD 9 codes 410-414 and ICD10 codes I20-I25). Direct health care costs were estimated from spending on prevention, accident and emergency care, hospital care, rehabilitation, and drug treatment. Direct non-health service costs were estimated from data on informal care. "Friction period" adjusted productivity costs were estimated using the human capital approach from lost earnings attributable to coronary heart disease related mortality and morbidity. The friction period is the period of employees' absence from work before the employer replaces them with other workers. Failure to adjust for this factor would overstate production loss. Coronary heart disease cost pound 1.73 billion to the UK health care system in 1999: pound 2.42 billion in informal care and pound 2.91 billion in friction period adjusted productivity loss; 24.1% of production losses were attributable to mortality and 75.9% to morbidity. The total annual cost of all coronary heart disease related burdens was pound 7.06 billion, the highest of all diseases in the UK for which comparable analyses have been done. Coronary heart disease is a leading public health problem in the UK in terms of the economic burden from disease. Cost estimates would be substantially understated if informal care/productivity costs were excluded.


Antibody levels to multiple malaria vaccine candidate antigens in relation to clinical malaria episodes in children in the Kasena-Nankana district of Northern Ghana.
Considering the natural history of malaria of continued susceptibility to infection and episodes of illness that decline in frequency and severity over time, studies which attempt to relate immune response to protection must be longitudinal and have clearly specified definitions of immune status. Putative vaccines are expected to protect against infection, mild or severe disease or reduce transmission, but so far it has not been easy to clearly establish what constitutes protective immunity or how this develops naturally, especially among the affected target groups. The present study was done in under six year old children to identify malaria antigens which induce antibodies that correlate with protection from Plasmodium falciparum malaria. In this longitudinal study, the multiplex assay was used to measure IgG antibody levels to 10 malaria antigens (GLURP R0, GLURP R2, MSP3 FVO, AMA1 FVO, AMA1 LR32, AMA1 3D7, MSP1 3D7, MSP1 FVO, LSA-1and EBA175RII) in 325 children aged 1 to 6 years in the Kassena Nankana district of northern Ghana. The antigen specific antibody levels were then related to the risk of clinical malaria over the ensuing year using a negative binomial regression model. IgG levels generally increased with age. The risk of clinical malaria decreased with increasing antibody levels. Except for FMPOII-LSA, (p = 0.05), higher IgG levels were associated with reduced risk of clinical malaria (defined as axillary temperature ≥37.5°C and parasitaemia of ≥5000 parasites/ul blood) in a univariate analysis, upon correcting for the confounding effect of age. However, in a combined multiple regression analysis, only IgG levels to MSP1-3D7 (Incidence rate ratio = 0.84, [95% C.I.= 0.73, 0.97, P = 0.02]) and AMA1 3D7 (IRR = 0.84 [95% C.I.= 0.74, 0.96, P = 0.01]) were associated with a reduced risk of clinical malaria over one year of morbidity surveillance. The data from this study support the view that a multivalent vaccine involving different antigens is most likely to be more effective than a monovalent one. Functional assays, like the parasite growth inhibition assay will be necessary to confirm if these associations reflect functional roles of antibodies to MSP1-3D7 and AMA1-3D7 in this population.