Long-Term Measurements of Human Inflammatory Cytokines Reveal Complex Baseline Variations between Individuals.
Comprehensive characterization of the healthy human proteome baseline is essential for personalized medicine. Baseline data are necessary to understand the variation between individuals, as well as longitudinal variation within individuals. Many important protein biomarkers, such as cytokines, exist at extremely low or undetectable levels in the healthy state. This paper describes results from a 14-week study of healthy human subjects using ultrasensitive single-molecule array (Simoa) assays to measure both intra and intersubject variation of 15 cytokines. The results show a wide variation in the ranges of some cytokines between individuals and demonstrate that individual baseline values will be essential for predicting disease presence and progression. Although all of the studied cytokines demonstrated high temporal stability (or low intrasubject variation) over the entire study period, there were two distinct groups of cytokines that demonstrated either high (IL-8, IFN-γ, IL-2, IL-6, and IL-1β) or low (IL-15, TNF-α, IL-12 p70, IL-17A, GM-CSF, IL-12 p40, IL-10, IL-7, IL-1α, and IL-5) subject-to-subject variation. This work demonstrates that ultrasensitive assays are essential for characterizing human cytokines in healthy subjects. The results show that some cytokines vary by more than two orders of magnitude between individuals, making it an imperative to obtain individual baseline measurements if they are to play a role in health and disease diagnosis.


Polymorphisms in the human monomethylarsonic acid (MMA V) reductase/hGSTO1 gene and changes in urinary arsenic profiles.
Large interindividual variability in urinary arsenic profiles, following chronic inorganic arsenic exposure, is well-known in humans. To understand this variability, we studied the relationship between polymorphisms in the gene for human monomethylarsonic acid (MMA(V)) reductase/hGSTO1 and the urinary arsenic profiles of individuals chronically exposed to arsenic in their drinking water. To ensure that we did not overlook rare polymorphisms, not included in the public databases, we amplified and sequenced all six exons of the gene and their flanking regions, using DNA isolated from peripheral blood samples of 75 subjects, living in the vicinity of Torreon, Mexico. Four groups, based on the levels of arsenic (9-100 microg/L) in their drinking water, were studied. We identified six novel polymorphisms and two reported previously. The novel polymorphisms were a three base pair deletion (delGGC) in the first intron; a G > C transversion, leading to a serine-to-cysteine substitution at amino acid 86; a G > T transversion and a A > T transversion in intron 5; a G > A transition resulting in glutamate-to-lysine substitution in amino acid 208; and a C > T transition producing an alanine-to-valine substitution in amino acid 236. Two subjects displayed significant differences in patterns of urinary arsenic; they had increased levels of urinary inorganic arsenic and reduced levels of methylated urinary arsenic species as compared to the rest of the study population. These two subjects had the same unique polymorphisms in hGSTO1 in that they were heterozygous for E155del and Glu208Lys. The identified SNPs may be one of the reasons for the large interindividual variability in the response of humans to chronic inorganic arsenic exposure. The findings suggest the need for further studies to identify unambiguously specific polymorphisms that may account for interindividual variability in the human response to chronic inorganic arsenic exposure.


Emotional Responses to Persistent Chemotherapy-induced Peripheral Neuropathy Experienced by Patients with Colorectal Cancer in Japan.
For patients who wish to continue treatment for persistent chemotherapy-induced peripheral neuropathy (PCIPN) while maintaining the quality of life (QOL), the only effective way appears to be the stop-and-go strategy. The objective of the present study was to analyze emotional responses of Japanese patients with colorectal cancer (CRC) experiencing PCIPN for the first time and to consider effective ways of providing supportive nursing care. In all, 25 patients with metastatic CRC who had completed more than six courses receiving first-line therapy with modified FOLFOX6 were included. Data were collected through semi-structured interviews. Emotional responses to PCIPN experienced by patients with CRC fell into the following categories: (1) suffering from an inability to cope with the previously unknown sensation of numbness, (2) inability to perform daily living activities activities of daily living [ADL]/instrumental activities of daily living [IADL] independently and feeling fear for physical safety, (3) feeling a sense of relief at being able to live the same life as before, and (4) facing the threat of cancer and tolerating the numbness caused by the life-supporting treatment. Fear, helplessness, dismay and other uncomfortable feelings represented 72.5% of all emotional responses, probably reflecting the lack of effective intervention for PCIPN. These results suggest that both subjective and objective assessments of PCIPN and proper use of a stop-and-go strategy are essential for treatment continuation and maintenance of patient's QOL; therefore, an integrated approach is desirable.


Breastfeeding Practices among Poor Women in Mesoamerica.
Breastfeeding is an effective intervention to reduce pediatric morbidity and mortality. The prevalence of practices and predictors of breastfeeding among the poor in Mesoamerica has not been well described. We estimated the prevalence of ever breastfeeding, early initiation of breastfeeding, exclusive breastfeeding, and breastfeeding between 6 mo and 2 y of age using household survey data for the poorest quintile of families living in 6 Mesoamerican countries. We also assessed the predictors of breastfeeding behaviors to identify factors amenable to policy interventions. We analyzed data from 12,529 children in Guatemala, Honduras, Mexico (Chiapas State), Nicaragua, Panama, and El Salvador using baseline survey data from the Salud Mesoamérica 2015 Initiative. We created multivariable Poisson regression models with robust variance estimates to calculate adjusted risk ratios (aRRs) and 95% CIs for breastfeeding outcomes and to control for sociodemographic and healthcare-related factors. Approximately 97% of women in all countries breastfed their child at least once, and 65.1% (Nicaragua) to 79.0% (Panama) continued to do so between 6 mo and 2 y of age. Breastfeeding in the first hour of life varied by country (P < 0.001), with the highest proportion reported in Panama (89.8%) and the lowest in El Salvador (65.6%). Exclusive breastfeeding also varied by country (P = 0.037), ranging from 44.5% in Panama to 76.8% in Guatemala. For every 20% increase in the proportion of peers who exclusively breastfed, there was an 11% (aRR: 1.11, 95% CI: 1.04, 1.18) increase in the likelihood of exclusive breastfeeding. Our study revealed significant variation in the prevalence of breastfeeding practices by poor women across countries surveyed by the Salud Mesoamérica 2015 initiative. Future interventions to promote exclusive breastfeeding should consider ways to leverage the role of the community in supporting individual women.


High frequency electromagnetic fields (GSM signals) affect gene expression levels in tumor suppressor p53-deficient embryonic stem cells.
Effects of electromagnetic fields (EMF) simulating exposure to the Global System for Mobile Communications (GSM) signals were studied using pluripotent embryonic stem (ES) cells in vitro. Wild-type ES cells and ES cells deficient for the tumor suppressor p53 were exposed to pulse modulated EMF at 1.71 GHz, lower end of the uplink band of GSM 1800, under standardized and controlled conditions, and transcripts of regulatory genes were analyzed during in vitro differentiation. Two dominant GSM modulation schemes (GSM-217 and GSM-Talk), which generate temporal changes between GSM-Basic (active during talking phases) and GSM-DTX (active during listening phases thus simulating a typical conversation), were applied to the cells at and below the basic safety limits for local exposures as defined for the general public by the International Commission on Nonionizing Radiation Protection (ICNIRP). GSM-217 EMF induced a significant upregulation of mRNA levels of the heat shock protein, hsp70 of p53-deficient ES cells differentiating in vitro, paralleled by a low and transient increase of c-jun, c-myc, and p21 levels in p53-deficient, but not in wild-type cells. No responses were observed in either cell type after EMF exposure to GSM-Talk applied at similar slot-averaged specific absorption rates (SAR), but at lower time-averaged SAR values. Cardiac differentiation and cell cycle characteristics were not affected in embryonic stem and embryonic carcinoma cells after exposure to GSM-217 EMF signals. Our data indicate that the genetic background determines cellular responses to GSM modulated EMF. Bioelectromagnetics 25:296-307, 2004.


Stabilized integrin-targeting ternary LPD (lipopolyplex) vectors for gene delivery designed to disassemble within the target cell.
Recent research in the field of nonviral gene delivery vectors has focused on preparing nanoparticles that are stabilized by the incorporation of a PEG coating and where one of the vector components is also cleavable. Here,we describe the synthesis, formulation, transfection properties, and biophysical studies of a PEG-stabilized ternary lipopolyplex vector in which, for the first time, both the lipid and peptide components are designed to be cleaved once the vector has been internalized. A series of cationic lipids, bearing short tri- or hexaethylene glycol groups, attached to the headgroup via an ester linkage, has been prepared. Trifunctional peptides have also been prepared, consisting of a Lys(16) sequence at the N-terminus (to bind and condense plasmid DNA); a spacer group (containing a sequence recognized and cleaved by endosomal enzymes) and an optional PEG4 amino acid; and an integrin-targeting cyclic peptide sequence (allowing the resulting nanoparticle to be internalized via receptor-mediated endocytosis). Differing combinations of these lipids and peptides have been formulated with DOPE and with plasmid DNA, and complex stability, transfection, and cleavage studies carried out. It was shown that optimal transfection activities in a range of cell types and complex stabilities were achieved with lipids bearing short cleavable triethylene glycol moieties, whereas the incorporation of PEG4 amino acids into the cleavable peptides had little effect. We have synthesized appropriate fluorescently labeled components and have studied the uptake of the vector, endosomal escape, peptide cleavage, and plasmid transport to the nucleus in breast cancer cells using confocal microscopy. We have also studied the morphology of these compact, stabilized vectors using cryo-EM.


Development of a PacBio Long-Read Sequencing Assay for High Throughput Detection of Fungicide Resistance in Zymoseptoria tritici.
Fungicide resistance has become a challenging problem in management of Septoria tritici blotch (STB), caused by Zymoseptoria tritici, the most destructive disease of winter wheat throughout western and northern Europe. To ensure the continued effectiveness of those fungicides currently used, it is essential to monitor the development and spread of such resistance in field populations of the pathogen. Since resistance to the key families of fungicides used for STB control (demethyalation inhibitors or azoles, succinate dehydrogenase inhibitors or SDHIs and Quinone outside Inhibitors or QoIs) is conferred through target-site mutations, the potential exists to monitor resistance through the molecular detection of alterations in the target site genes. As more efficient fungicides were developed and applied, the pathogen has continuously adapted through accumulating multiple target-site alterations. In order to accurately monitor these changes in field populations, it is therefore becoming increasingly important to completely sequence the targeted genes. Here we report the development of a PacBio assay that facilitates the multiplex amplification and long-read sequencing of the target gene(s) for the azole (CYP51), SDHI (Sdh B, C, and D), and QoI (cytochrome b) fungicides. The assay was developed and optimised using three Irish Z. tritici collections established in spring 2017, which capture the range of fungicide resistance present in modern European populations of Z. tritici. The sequences obtained through the PacBio assay were validated using traditional Sanger sequencing and in vitro sensitivity screenings. To further exploit the long-read and high throughput potential of PacBio sequencing, an additional nine housekeeping genes (act, BTUB, cal, cyp, EF1, GAPDH, hsp80-1, PKC, TFC1) were sequenced and used to provide comprehensive Z. tritici strain genotyping.


Can community care workers deliver a falls prevention exercise program? A feasibility study.
Almost half of older people receiving community care fall each year and this rate has not improved in the last decade. Falls prevention programs targeted at this group are uncommon, and expensively delivered by university trained allied health professionals. To investigate the feasibility of community care workers delivering a falls prevention exercise program to older clients, at low or medium risk of falling, as part of an existing service provision. Community care workers from 10 community care organizations participated in the training for, and delivery to their clients of, an 8-week evidence-based falls prevention exercise program. Community care workers included assessment staff (responsible for identifying the need for community care services through completing an assessment) and support workers (responsible for providing support in the home). Clients were surveyed anonymously at the completion of the intervention and workers participated in a semi-structured interview. Twenty-five community care workers participated in the study. The falls prevention program was delivered to 29 clients, with an average age of 82.7 (SD: 8.72) years and consisting of 65.5% female. The intervention was delivered safely with no adverse events recorded, and the eligibility and assessment tools were completed by the majority of community care workers (93.1%). Assessment staff found it difficult to find time to deliver the intervention. Support workers were able to complete the intervention within their current service delivery period, with the initial assessment taking a small amount of additional time. Support workers reported enjoying the additional responsibility afforded by delivering the falls prevention program and seeing changes in their clients. The majority of clients (82%) reported enjoying the exercises, with 59% reporting that they felt it made a positive change in their health. Clients completed the exercises on average 4.8 (SD: 2.2) days per week. Community care workers who have completed appropriate training are able to deliver a falls prevention exercise program to their clients as part of their current services. Further research is required to determine whether the program reduces the rate of falls for community care clients and whether integration of a falls prevention program into an existing service is cost-effective.


Aerosol infection of mice with Bordetella pertussis.
Aerosol inhalation of Bordetella pertussis Tohama phase I resulted in a reproducible and uniform infection of mice (strain DDY or ICR). Mice in groups of 10 exposed for 30 min to aerosols generated from bacterial suspensions of 10(9) and 10(10) organisms per ml resulted in mean bacterial counts of 2.3 (+/- 0.3) X 10(4) and 1.0 (+/- 0.3) X 10(5) colony-forming units, respectively, in the lung of each animal. Subsequent studies using a 30-min aerosol inoculation of ICR mice with 2 X 10(9) bacterial cells per ml showed: (i) B. pertussis cells reached a maximum of about 10(7) colony-forming units per lung 14 days after inhalation. (ii) Deaths (10 to 100%, depending on mouse age) occurred 10 to 14 days after exposure. (iii) The lung weight and the leukocyte count increased from basal values of 100 mg and 10(4) leukocytes per mm3 to a plateau of 950 mg and 1.95 X 10(5) leukocytes per mm3, respectively, 14 days after challenge. (iv) There was a significantly reduced rate of body weight gain by infected mice compared to noninfected mice. (v) With mortality as the criterion for disease, susceptibility varied with the age of mice as follows: 10 days old greater than 18 greater than 28 greater than 49. (vi) Bacteria were associated with ciliated respiratory epithelial cells by scanning electron microscopy.


Overcoming knowledge stickiness in scientific knowledge transfer.
This paper explores the transfer and dissemination of knowledge between scientists, the volunteers who collect the knowledge and the communities which learn from it in order to implement change. The role of knowledge "stickiness" in the reduction of knowledge transfer is outlined. The characteristics of the knowledge and the situation combine to develop a range of factors, "stickiness predictors," which can deter knowledge transfer. These stickiness predictors are used to analyse data gathered from three qualitative cases, which were developed from both participant observation and semi-structured interviews studying the interactions between the scientists, volunteers and organisations. A reconsideration of the way that knowledge and knowledge transfer are being conceptualised by scientists is proposed, in order to enable "stickiness" factors to be recognised and managed, thereby increasing the potential for scientific literacy. A move towards a more broadly constituted community of practice is proposed.


A novel method to compensate for different amplification efficiencies between patient DNA samples in quantitative real-time PCR.
Quantification of residual disease by real-time polymerase chain reaction (PCR) will become a pivotal tool in the development of patient-directed therapy. In recent years, various protocols to quantify minimal residual disease in leukemia or lymphoma patients have been developed. These assays assume that PCR efficiencies are equal for all samples. Determining t(14;18) and albumin reaction efficiencies for sixteen follicular lymphoma patient samples revealed higher efficiencies for blood samples than for lymph node samples in general. However, within one sample both reactions had equivalent efficiencies. Differences in amplification efficiencies between patient samples (low efficiencies) and the calibrator in quantitative analyses result in the underestimation of residual disease in patient samples whereby the weakest positive patient samples are at highest error. Based on these findings for patient samples, the efficiency compensation control was developed. This control includes two reference reactions in a multiplex setting, specific for the beta-actin and albumin housekeeping genes that are present in a constant ratio within DNA templates. The difference in threshold cycle values for both reference reactions, ie, the Ct(2) value, is dependent on the amplification efficiency, and is used to compensate for efficiency differences between patient samples and the calibrator. The beta-actin reference reaction is also used to normalize for DNA input. Furthermore, the efficiency compensation control facilitates identification of patient samples that are so contaminated with PCR inhibitory compounds that different amplification reactions are affected to a different extent. Accurate quantitation of residual disease in these samples is therefore impossible with the current quantitative real-time PCR protocols. Identification and exclusion of these inadequate samples will be of utmost importance in quantitative retrospective studies, but even more so, in future molecular diagnostic analyses.


Use of potentially inappropriate medications among older outpatients and inpatients in a tertiary care hospital in Malaysia.
Older individuals are seemingly having more medical conditions, which predispose them to a greater risk of polypharmacy. Potentially inappropriate medications (PIMs), including those having anticholinergic and sedative properties, are common in their prescriptions, often associated with functional decline and negative health outcomes. Thus, this study reports proportions of inappropriate drugs and drug burden exposures and its correlation with patient-reported outcomes (PROs) among cognitively intact older adults admitted to a ward or visiting the outpatient clinic at a tertiary care hospital in Malaysia. This cross-sectional study included data from 344 older (173 inpatients and 171 outpatients) patients, aged 60 years and above, through validated questionnaires. Medication appropriateness was assessed via Medication Appropriateness Index (MAI) tool, whereas Beers and Screening Tool of Older Person's Potentially Inappropriate Prescribing (STOPP) criteria were used to evaluate PIMs and potentially inappropriate prescribing (PIP), respectively. The Drug Burden Index (DBI) and polypharmacy, as well as PROs, included Groningen Frailty Indicator (GFI), Katz Index of Independence in Activities of Daily Living (Katz ADL) and Older People's Quality of Life (OPQOL) were also evaluated. Overall, inpatients received significantly higher medications (6.90 ± 2.70 vs 4.49 ± 3.20) than outpatients. A significantly higher proportion of inpatients received at least one PIM (65% vs 57%) or PIP (57.4% vs 17.0%) and higher mean MAI score (1.76 ± 1.08 and 1.10 ± 0.34) and DBI score (2.67 ± 1.28 vs 1.49 ± 1.17) than outpatients. Inpatients had significantly higher total OPQOL (118.53 vs 79.95) and GFI score (5.44 vs 3.78) than outpatients. We only found significant correlations between GFI and DBI and total OPQOL and the number of PIMs. Proportions of PIMs and DBI exposure were significantly higher in an inpatient setting. No significant correlations between exposures to inappropriate medications or drug burden and PROs were observed.


A three-dimensional study of sexual dimorphism in the human face.
The sexual dimorphism in three-dimensional facial form (size plus shape) was investigated in a sample of 40 men and 36 women by using Euclidean-distance matrix analysis. Subjects ranged in age from 19 to 32 years, had excellent dentitions, and had no craniocervical disorders. For each subject, 16 facial landmarks were automatically collected using a computerized system consisting of two infrared CCD cameras, real-time hardware for the recognition of markers, and software for the three-dimensional reconstruction of landmarks' x, y, z coordinates. Euclidean-distance matrix analysis confirmed the well-known size difference between adult male and female faces (men's faces being 6% to 7% larger than women's faces), while it demonstrated no significant gender differences in three-dimensional facial shape. This result contrasted with the shape differences previously found when separate two-dimensional frontal and sagittal plane projections were analyzed. It could be explained by a relative three-dimensional compensation between the different facial dimensions.


Virotherapy in Germany-Recent Activities in Virus Engineering, Preclinical Development, and Clinical Studies.
Virotherapy research involves the development, exploration, and application of oncolytic viruses that combine direct killing of cancer cells by viral infection, replication, and spread (oncolysis) with indirect killing by induction of anti-tumor immune responses. Oncolytic viruses can also be engineered to genetically deliver therapeutic proteins for direct or indirect cancer cell killing. In this review-as part of the special edition on "State-of-the-Art Viral Vector Gene Therapy in Germany"-the German community of virotherapists provides an overview of their recent research activities that cover endeavors from screening and engineering viruses as oncolytic cancer therapeutics to their clinical translation in investigator-initiated and sponsored multi-center trials. Preclinical research explores multiple viral platforms, including new isolates, serotypes, or fitness mutants, and pursues unique approaches to engineer them towards increased safety, shielded or targeted delivery, selective or enhanced replication, improved immune activation, delivery of therapeutic proteins or RNA, and redirecting antiviral immunity for cancer cell killing. Moreover, several oncolytic virus-based combination therapies are under investigation. Clinical trials in Germany explore the safety and potency of virotherapeutics based on parvo-, vaccinia, herpes, measles, reo-, adeno-, vesicular stomatitis, and coxsackie viruses, including viruses encoding therapeutic proteins or combinations with immune checkpoint inhibitors. These research advances represent exciting vantage points for future endeavors of the German virotherapy community collectively aimed at the implementation of effective virotherapeutics in clinical oncology.


The Orphan Cytokine Receptor CRLF3 Emerged With the Origin of the Nervous System and Is a Neuroprotective Erythropoietin Receptor in Locusts.
The orphan cytokine receptor-like factor 3 (CRLF3) was identified as a neuroprotective erythropoietin receptor in locust neurons and emerged with the evolution of the eumetazoan nervous system. Human CRLF3 belongs to class I helical cytokine receptors that mediate pleiotropic cellular reactions to injury and diverse physiological challenges. It is expressed in various tissues including the central nervous system but its ligand remains unidentified. A CRLF3 ortholog in the holometabolous beetle Tribolium castaneum was recently shown to induce anti-apoptotic mechanisms upon stimulation with human recombinant erythropoietin. To test the hypothesis that CRLF3 represents an ancient cell-protective receptor for erythropoietin-like cytokines, we investigated its presence across metazoan species. Furthermore, we examined CRLF3 expression and function in the hemimetabolous insect Locusta migratoria. Phylogenetic analysis of CRLF3 sequences indicated that CRLF3 is absent in Porifera, Placozoa and Ctenophora, all lacking the traditional nervous system. However, it is present in all major eumetazoan groups ranging from cnidarians over protostomians to mammals. The CRLF3 sequence is highly conserved and abundant amongst vertebrates. In contrast, relatively few invertebrates express CRLF3 and these sequences show greater variability, suggesting frequent loss due to low functional importance. In L. migratoria, we identified the transcript Lm-crlf3 by RACE-PCR and detected its expression in locust brain, skeletal muscle and hemocytes. These findings correspond to the ubiquitous expression of crlf3 in mammalian tissues. We demonstrate that the sole addition of double-stranded RNA to the culture medium (called soaking RNA interference) specifically interferes with protein expression in locust primary brain cell cultures. This technique was used to knock down Lm-crlf3 expression and to abolish its physiological function. We confirmed that recombinant human erythropoietin rescues locust brain neurons from hypoxia-induced apoptosis and showed that this neuroprotective effect is absent after knocking down Lm-crlf3. Our results affirm the erythropoietin-induced neuroprotective function of CRLF3 in a second insect species from a different taxonomic group. They suggest that the phylogenetically conserved CRLF3 receptor may function as a cell protective receptor for erythropoietin or a structurally related cytokine also in other animals including vertebrate and mammalian species.


Consideration of the efficacy of non-ionic vesicles in the targeted delivery of oral vaccines.
The fundamentals of this research were to exploit non-ionic surfactant technology for delivery and administration of vaccine antigens across the oral route and to gain a better understanding of vaccine trafficking. Using a newly developed method for manufacture of non-ionic surfactant vesicles (niosomes and bilosomes) lower process temperatures were adopted thus reducing antigen exposure to potentially damaging conditions. Vesicles prepared by this method offered high protection to enzymatic degradation, with only ∼10 % antigen loss measured when vesicles incorporating antigen were exposed to enzyme digestion. Interestingly, when formulated using this new production method, the addition of bile salt to the vesicles offered no advantage in terms of stability within simulated gastro-intestinal conditions. Considering their ability to deliver antigen to their target site, results demonstrated that incorporation of antigen within vesicles enhanced delivery and targeting of the antigen to the Peyer's Patch, again with niosomes and bilosomes offering similar efficiency. Delivery to both the Peyer's patches and mesentery lymphatics was shown to be dose dependent at lower concentrations, with saturation kinetics applying at higher concentrations. This demonstrates that in the formulation of vaccine delivery systems, the lipid/antigen dose ratio is not only a key factor in production cost, but is equally a key factor in the kinetics of delivery and targeting of a vaccine system.


Effects of guided imagery on outcomes of pain, functional status, and self-efficacy in persons diagnosed with fibromyalgia.
(1) To investigate the effects of a 6-week intervention of guided imagery on pain level, functional status, and self-efficacy in persons with fibromyalgia (FM); and (2) to explore the dose-response effect of imagery use on outcomes. Longitudinal, prospective, two-group, randomized, controlled clinical trial. The sample included 48 persons with FM recruited from physicians' offices and clinics in the mid-Atlantic region. Participants randomized to Guided Imagery (GI) plus Usual Care intervention group received a set of three audiotaped guided imagery scripts and were instructed to use at least one tape daily for 6 weeks and report weekly frequency of use (dosage). Participants assigned to the Usual Care alone group submitted weekly report forms on usual care. All participants completed the Short-Form McGill Pain Questionnaire (SF-MPQ), Arthritis Self- Efficacy Scale (ASES), and Fibromyalgia Impact Questionnaire (FIQ), at baseline, 6, and 10 weeks, and submitted frequency of use report forms. FIQ scores decreased over time in the GI group compared to the Usual Care group (p = 0.03). Ratings of self-efficacy for managing pain (p = 0.03) and other symptoms of FM also increased significantly over time (p = < 0.01) in the GI group compared to the Usual Care group. Pain as measured by the SF-MPQ did not change over time or by group. Imagery dosage was not significant. This study demonstrated the effectiveness of guided imagery in improving functional status and sense of self-efficacy for managing pain and other symptoms of FM. However, participants' reports of pain did not change. Further studies investigating the effects of mind-body interventions as adjunctive self-care modalities are warranted in the fibromyalgia patient population.


Was rurality protective in the 1918 influenza pandemic in New Zealand?
This study aimed to examine the impact of rurality on mortality rates from pandemic influenza in New Zealand in 1918. Mortality data was obtained from death certificates (in a published source) and denominator population data from the 1916 census (for the European population only). Analyses were conducted on cities (n = 4), towns (n = 111), counties (n = 97). The influenza mortality rate for the towns and cities was more than twice that of the counties that represented rural settings (rate ratio (RR) = 2.13, 95% CI = 2.00-2.27). However, larger towns (population >2000 people) had a significantly lower mortality rate than smaller towns (RR = 0.81, 95%CI = 0.74-0.88). Similarly, cities had a lower mortality rate than larger towns (RR = 0.89, 95%CI = 0.83-0.95). These results are suggestive that rurality may have provided some protection from mortality during this influenza pandemic. This may have been due to a mix of remoteness and greater social distancing among rural residents. However, the differences in mortality rates between towns and cities may have reflected other factors such as the more organised provision of community care in the larger towns and cities, when compared to smaller towns.


Processive movement of single kinesins on crowded microtubules visualized using quantum dots.
Kinesin-1 is a processive molecular motor transporting cargo along microtubules. Inside cells, several motors and microtubule-associated proteins compete for binding to microtubules. Therefore, the question arises how processive movement of kinesin-1 is affected by crowding on the microtubule. Here we use total internal reflection fluorescence microscopy to image in vitro the runs of single quantum dot-labelled kinesins on crowded microtubules under steady-state conditions and to measure the degree of crowding on a microtubule at steady-state. We find that the runs of kinesins are little affected by high kinesin densities on a microtubule. However, the presence of high densities of a mutant kinesin that is not able to step efficiently reduces the average speed of wild-type kinesin, while hardly changing its processivity. This indicates that kinesin waits in a strongly bound state on the microtubule when encountering an obstacle until the obstacle unbinds and frees the binding site for kinesin's next step. A simple kinetic model can explain quantitatively the behaviour of kinesin under both crowding conditions.


Genetic heterogeneity in uveal melanoma assessed by multiplex ligation-dependent probe amplification.
To determine intratumor genetic heterogeneity in uveal melanoma (UM) by multiplex ligation-dependent probe amplification (MLPA) in formalin-fixed, paraffin-embedded (FFPE) tumor tissues. DNA was extracted from whole tumor sections and from two to nine different areas microdissected from 32 FFPE UMs. Thirty-one loci on chromosomes 1, 3, 6, and 8 were tested with MLPA for copy number changes. The tumor was considered heterogeneous at a locus if (1) the difference in dosage quotients (DQs) of any two areas was 0.2 or more, and (2) the DQs of the areas belonged to different ranges. Comparison of MLPA data obtained from microdissected areas of the UMs showed heterogeneity in 1 to 26 examined loci in 24 (75%) tumors, with only 25% of the tumors being homogeneous. Intratumor heterogeneity of 3p12.2, 6p21.2, and 8q11.23 was most common, occurring in >30% of the UMs. Gains of chromosome 3 were observed in four UMs, with three of these tumors showing the highest degree of heterogeneity. Copy number variation was associated with differences in tumor cell type, but not with differences in tumor pigmentation or reactive inflammation. UMs with genetic heterogeneity across multiple sample sites showed equivocal MLPA results when the whole tumor section was examined. These results suggest that different clones dilute MLPA results. Heterogeneity of chromosomal abnormalities of chromosomes 1, 3, 6, and 8 is present in most UMs. This heterogeneity causes equivocal MLPA results. One random tumor sample may not be representative of the whole tumor and, therefore, may be insufficient for prognostic testing.


The BTB-zinc finger transcription factor abrupt acts as an epithelial oncogene in Drosophila melanogaster through maintaining a progenitor-like cell state.
The capacity of tumour cells to maintain continual overgrowth potential has been linked to the commandeering of normal self-renewal pathways. Using an epithelial cancer model in Drosophila melanogaster, we carried out an overexpression screen for oncogenes capable of cooperating with the loss of the epithelial apico-basal cell polarity regulator, scribbled (scrib), and identified the cell fate regulator, Abrupt, a BTB-zinc finger protein. Abrupt overexpression alone is insufficient to transform cells, but in cooperation with scrib loss of function, Abrupt promotes the formation of massive tumours in the eye/antennal disc. The steroid hormone receptor coactivator, Taiman (a homologue of SRC3/AIB1), is known to associate with Abrupt, and Taiman overexpression also drives tumour formation in cooperation with the loss of Scrib. Expression arrays and ChIP-Seq indicates that Abrupt overexpression represses a large number of genes, including steroid hormone-response genes and multiple cell fate regulators, thereby maintaining cells within an epithelial progenitor-like state. The progenitor-like state is characterised by the failure to express the conserved Eyes absent/Dachshund regulatory complex in the eye disc, and in the antennal disc by the failure to express cell fate regulators that define the temporal elaboration of the appendage along the proximo-distal axis downstream of Distalless. Loss of scrib promotes cooperation with Abrupt through impaired Hippo signalling, which is required and sufficient for cooperative overgrowth with Abrupt, and JNK (Jun kinase) signalling, which is required for tumour cell migration/invasion but not overgrowth. These results thus identify a novel cooperating oncogene, identify mammalian family members of which are also known oncogenes, and demonstrate that epithelial tumours in Drosophila can be characterised by the maintenance of a progenitor-like state.


Electric-field dependence of the effective dielectric constant in graphene.
The dielectric constant of a material is one of the fundamental features used to characterize its electrostatic properties such as capacitance, charge screening, and energy storage capability. Graphene is a material with unique behavior due to its gapless electronic structure and linear dispersion near the Fermi level, which can lead to a tunable band gap in bilayer and trilayer graphene, a superconducting-insulating transition in hybrid systems driven by electric fields, and gate-controlled surface plasmons. All of these results suggest a strong interplay between graphene properties and external electric fields. Here we address the issue of the effective dielectric constant (ε) in N-layer graphene subjected to out-of-plane (E(ext)(⊥)) and in-plane (E(ext)(||)) external electric fields. The value of ε has attracted interest due to contradictory reports from theoretical and experimental studies. Through extensive first-principles electronic structure calculations, including van der Waals interactions, we show that both the out-of-plane (ε(⊥)) and the in-plane (ε(||)) dielectric constants depend on the value of applied field. For example, ε(⊥) and ε(||) are nearly constant (~3 and ~1.8, respectively) at low fields (E(ext) < 0.01 V/Å) but increase at higher fields to values that are dependent on the system size. The increase of the external field perpendicular to the graphene layers beyond a critical value can drive the system to a unstable state where the graphene layers are decoupled and can be easily separated. The observed dependence of ε(⊥) and ε(||) on the external field is due to charge polarization driven by the bias. Our results point to a promising way of understanding and controlling the screening properties of few-layer graphene through external electric fields.


Differential experiences of the psychobiological sequelae of ecstasy use: quantitative and qualitative data from an internet study.
Previous work provided preliminary evidence that different patterns of use among ecstasy users may impact on perceived side-effects. Participants recruited via an ecstasy-related bulletin board differed in their responses compared to those recruited via other means. The present investigation compares self-reports of psychobiological difficulties among ecstasy users recruited either via a bulletin board or by alternative methods. Qualitative data included reports of any negative or positive changes attributable to ecstasy use and reasons for cessation of use. An Internet-based design was utilized and 209 volunteers completed the study, 117 of whom were recruited via a bulletin board devoted to discussion of ecstasy. Psychobiological difficulties attributable to ecstasy use varied, with mood fluctuation the most common. Differences between the two groups in the extent to which these problems were reported was found. Bulletin board recruits were less likely to report anxiety or poor concentration, but more likely to report tremors/twitches. For the whole sample, lifetime use was associated more with psychobiologial problems, although this pattern was stronger and more pervasive for the non-bulletin board participants. Bulletin board recruits were more aware of possible negative psychological effects and were more likely to report adopting harm reduction strategies. From the qualitative data three negative consequences of use were identified, the most common of which was "psychological problems". In support of the quantitative findings the likelihood of reporting psychological problems increased with lifetime exposure to ecstasy in both recruitment conditions but interestingly this did not appear to impact on reasons for cessation of use. Participants also reported a number of effects that they regarded as beneficial. Future research should also take these aspects of use into account.


Retinal arterial macroaneurysms: risk factors of poor visual outcome.
To evaluate the long-term natural course and to identify the risk factors of a poor visual outcome in patients with retinal arterial macroaneurysms (MA). In this observational case series, we retrospectively evaluated the consecutive patients with MA at our institution over a 17-year period. The visual outcomes of the different types of MA (presented as predominantly hemorrhagic or exudative) were compared. A total of 31 patients with 46 confirmed MA were identified. The ocular manifestations of MA included retinal hemorrhage (96.7%), retinal exudate (61.3%), macular involvement (77.4%) and vitreous hemorrhage (12.9%). Among the 10 cases of hemorrhagic-type MA, visual acuity improved by 2 or more lines in 9 (90%) and decreased in 1 patient (10%). In the 15 cases of exudative-type MA, 4 improved (26.7%), 8 remained stationary (53.3%) and 3 had decreased visual acuity (20.0%). The average vision change in the minimum angle of resolution improved by 0.59 log units in hemorrhagic MA and decreased by 0.07 log units in exudative MA (p < 0.01). With respect to change of vision, the eyes with hemorrhagic-type MA showed greater improvement than those with the exudative type, but the final visual outcome was not different in the 2 groups. Foveal exudate was a statistically significant risk factor for final vision equal to or worse than 6/30 (odds ratio = 6.93, p < 0.05). Different types of MA may have different clinical presentations. In the majority of MA, there is gradual and spontaneous involution. However, poor visual outcome may occur secondary to foveal exudates and subfoveal hemorrhage.


Characterization of an interferon-induced 48-kD protein immunologically related to the double-stranded RNA-activated protein kinase PKR.
Polyclonal antibodies raised against purified and urea-denatured double-stranded protein kinase (PKR) from human origin cross-reacted by immunoblotting with a 48-kD protein (p48) induced by the three types of interferon (IFN), alpha, beta, and gamma. The induction of p48 is IFN dose dependent and its accumulation occurs a few hours after the addition of IFN. The induction of p48 is blocked by actinomycin D. Analysis by two-dimensional gel isoelectric-focusing, revealed p48 as a single spot with an isoelectric point (pI) of 6.8. In the same experiment the PKR was revealed as several subspecies with pI values in the pH range of 7.4-8.0. Cell fractionation experiments indicated that PKR and p48 have different subcellular localizations: PKR was found to be associated with the microsomal pellet as shown previously whereas p48 was recovered in the microsomal supernatant fraction. In addition to these differences, PKR and p48 were found to be differentially expressed in some human cells treated with the three types of IFN. For example, in HeLa cells, IFN-alpha or IFN-beta induced similarly both PKR and p48 whereas IFN-gamma induced mainly p48. In U937 cells in which PKR was not expressed with or without IFN treatment, p48 was strongly induced by all three types of IFN. These results suggest different mechanisms for the induction of PKR and p48. In view of its presence in different types of human cells and its induction by different types of IFN, it is possible to suggest that p48 might play an important role in mediating some of the action of IFN.


Micro- and minisatellite-expressed sequence tag (EST) markers discriminate between populations of Rhipicephalus appendiculatus.
Biological differences, including vector competence for the protozoan parasite Theileria parva have been reported among populations of Rhipicephalus appendiculatus (Acari: Ixodidae) from different geographic regions. However, the genetic diversity and population structure of this important tick vector remain unknown due to the absence of appropriate genetic markers. Here, we describe the development and evaluation of a panel of EST micro- and minisatellite markers to characterize the genetic diversity within and between populations of R. appendiculatus and other rhipicephaline species. Sixty-six micro- and minisatellite markers were identified through analysis of the R. appendiculatus Gene Index (RaGI) EST database and selected bacterial artificial chromosome (BAC) sequences. These were used to genotype 979 individual ticks from 10 field populations, 10 laboratory-bred stocks, and 5 additional Rhipicephalus species. Twenty-nine markers were polymorphic and therefore informative for genetic studies while 6 were monomorphic. Primers designed from the remaining 31 loci did not reliably generate amplicons. The 29 polymorphic markers discriminated populations of R. appendiculatus and also 4 other Rhipicephalus species, but not R. zambeziensis. The percentage Principal Component Analysis (PCA) implemented using Multiple Co-inertia Analysis (MCoA) clustered populations of R. appendiculatus into 2 groups. Individual markers however differed in their ability to generate the reference typology using the MCoA approach. This indicates that different panels of markers may be required for different applications. The 29 informative polymorphic micro- and minisatellite markers are the first available tools for the analysis of the phylogeography and population genetics of R. appendiculatus.


Child sexual abuse in religiously affiliated and secular institutions: a retrospective descriptive analysis of data provided by victims in a government-sponsored reappraisal program in Germany.
The disclosure of widespread sexual abuse committed by professional educators and clergymen in institutions in Germany ignited a national political debate, in which special attention was paid to church-run institutions. We wanted to find out whether the nature of the abuse and its effect on victims differed depending on whether the abuse had been experienced in religiously affiliated versus secular institutions. In 2010, the German government established a hotline that victims could contact anonymously to describe their experiences of sexual abuse. The information provided by callers was documented and categorized. Our analysis looked at a subset of the data collected, in order to compare the nature of the abuse experienced at three types of institutions: Roman Catholic, Protestant, and non-religiously affiliated. Non-parametric tests were used to compare frequency distributions, and qualitative data were analyzed descriptively. Of the 1050 victims in our sample, 404 had been in Roman Catholic, 130 in Protestant, and 516 in non-religious institutions. The overall mean age at the time of reporting was 52.2 years. Males (59.8%) outnumbered females. Victims who had been in religiously affiliated institutions were significantly older than those who had been in secular institutions. Almost half the victims had been abused physically as well as sexually, and most victims reported that the abuse had occurred repeatedly and that the assaults had been committed by males. Patterns of abuse (time, type, and extent), and the gender of the offenders did not differ between the three groups. Intercourse was more frequently reported by older victims and by females. Similar percentages of victims in all groups reported current psychiatric diagnoses (depression, anxiety disorders, PTSD). Significantly more victims from Protestant institutions reported having current psychosocial problems. The results suggest that child sexual abuse in institutions is attributable to the nature of institutional structures and to societal assumptions about the rights of children more than to the attitudes towards sexuality of a specific religion. The exploratory data arising from this study may serve as a starting point for building hypotheses, and may point the way toward improvements in prevention and intervention strategies.


A broad autism phenotype expressed in facial morphology.
Autism spectrum disorder is a heritable neurodevelopmental condition diagnosed based on social and communication differences. There is strong evidence that cognitive and behavioural changes associated with clinical autism aggregate with biological relatives but in milder form, commonly referred to as the 'broad autism phenotype'. The present study builds on our previous findings of increased facial masculinity in autistic children (Sci. Rep., 7:9348, 2017) by examining whether facial masculinity represents as a broad autism phenotype in 55 non-autistic siblings (25 girls) of autistic children. Using 3D facial photogrammetry and age-matched control groups of children without a family history of ASD, we found that facial features of male siblings were more masculine than those of male controls (n = 69; p < 0.001, d = 0.81 [0.36, 1.26]). Facial features of female siblings were also more masculine than the features of female controls (n = 60; p = 0.005, d = 0.63 [0.16, 1.10]). Overall, we demonstrated for males and females that facial masculinity in non-autistic siblings is increased compared to same-sex comparison groups. These data provide the first evidence for a broad autism phenotype expressed in a physical characteristic, which has wider implications for our understanding of the interplay between physical and cognitive development in humans.


The role of temperature in determining species' vulnerability to ocean acidification: a case study using Mytilus galloprovincialis.
Ocean acidification (OA) is occurring across a backdrop of concurrent environmental changes that may in turn influence species' responses to OA. Temperature affects many fundamental biological processes and governs key reactions in the seawater carbonate system. It therefore has the potential to offset or exacerbate the effects of OA. While initial studies have examined the combined impacts of warming and OA for a narrow range of climate change scenarios, our mechanistic understanding of the interactive effects of temperature and OA remains limited. Here, we use the blue mussel, Mytilus galloprovincialis, as a model species to test how OA affects the growth of a calcifying invertebrate across a wide range of temperatures encompassing their thermal optimum. Mussels were exposed in the laboratory to a factorial combination of low and high pCO2 (400 and 1200 µatm CO2) and temperatures (12, 14, 16, 18, 20, and 24°C) for one month. Results indicate that the effects of OA on shell growth are highly dependent on temperature. Although high CO2 significantly reduced mussel growth at 14°C, this effect gradually lessened with successive warming to 20°C, illustrating how moderate warming can mediate the effects of OA through temperature's effects on both physiology and seawater geochemistry. Furthermore, the mussels grew thicker shells in warmer conditions independent of CO2 treatment. Together, these results highlight the importance of considering the physiological and geochemical interactions between temperature and carbonate chemistry when interpreting species' vulnerability to OA.


Silicon nanocrystals on bottle resonators: mode structure, loss mechanisms and emission dynamics.
Bottle resonators can support high Q-factor whispering gallery modes (WGMs) and demonstrate a rich mode spectrum. Resonators were fabricated using a fiber tapering apparatus and were coated with a thin, smooth layer of luminescent silicon nanocrystals. The photoluminescence spectrum showed WGM peaks with Q-factors near 2,500; however, evanescent measurements showed that these modes are a composite of many modes with Q-factors exceeding 106, the highest yet seen for a silicon-nanocrystalcoated microresonators. The mode structure showed strong polarization and sensitivity to position within the bottle resonator. An analysis of loss mechanisms establishes surface roughness scattering as the limiting factor in these nanocrystal-coated bottle resonators in the absence of excited carriers.


Making arm movements within different parts of space: the premotor and motor cortical representation of a coordinate system for reaching to visual targets.
The activity of 156 individual arm-related neurons was studied in the premotor cortex (area 6) while monkeys made arm movements of similar directions within different parts of 3-dimensional space. This study was aimed at describing the relationship between premotor cortical cell activity and direction of arm movement and assessing the coordinate system underlying this relationship. We found that the activity of 152 (97.4%) cells varied in an orderly fashion with the direction of movement, in at least some region of the work space. Premotor cortical cells fired most for a given preferred direction and less for other directions of movement. These preferred directions covered the directional continuum in a uniform fashion across the work space. It was found that, as movements of similar directions were made within different parts of the work space, the cells' preferred directions changed their orientation. Although these changes had different magnitudes for different cells, at the population level, they followed closely the changes in orientation of the arm necessary to move the hand from one to another part of the work space. This shift of cells' preferred direction with the orientation of the arm in space has been observed with similar characteristics in the motor cortex (see Caminiti et al., 1990). In both premotor and motor cortices, neuronal movement population vectors provide a good description of movement direction. Unlike the individual cell preferred directions upon which they are based, movement population vectors did not change their spatial orientation across the work space, suggesting that they remain good predictors of movement direction regardless of the region of space in which movements are made. The firing frequency of both premotor and motor cortical neurons varied significantly with the position occupied by the hand in space. These static positional effects were observed in 88.5% of premotor and 91.8% of motor cortical cells. In a second task, monkeys made movements from differing origins to a common end point. This task was performed within 3 different parts of space and was aimed at dissociating movement direction from movement end point. It was found that in both premotor and motor cortices virtually all cells were related to the direction and not to the end point of movement. These data suggest that premotor and motor cortices use common mechanisms for coding arm movement direction. They also provide a basis for understanding the coordinate transformation required to move the hand toward visual targets in space.


Reconciling the role of serotonin in behavioral inhibition and aversion: acute tryptophan depletion abolishes punishment-induced inhibition in humans.
The neuromodulator serotonin has been implicated in a large number of affective and executive functions, but its precise contribution to motivation remains unclear. One influential hypothesis has implicated serotonin in aversive processing; another has proposed a more general role for serotonin in behavioral inhibition. Because behavioral inhibition is a prepotent reaction to aversive outcomes, it has been a challenge to reconcile these two accounts. Here, we show that serotonin is critical for punishment-induced inhibition but not overall motor response inhibition or reporting aversive outcomes. We used acute tryptophan depletion to temporarily lower brain serotonin in healthy human volunteers as they completed a novel task designed to obtain separate measures of motor response inhibition, punishment-induced inhibition, and sensitivity to aversive outcomes. After a placebo treatment, participants were slower to respond under punishment conditions compared with reward conditions. Tryptophan depletion abolished this punishment-induced inhibition without affecting overall motor response inhibition or the ability to adjust response bias in line with punishment contingencies. The magnitude of reduction in punishment-induced inhibition depended on the degree to which tryptophan depletion reduced plasma tryptophan levels. These findings extend and clarify previous research on the role of serotonin in aversive processing and behavioral inhibition and fit with current theorizing on the involvement of serotonin in predicting aversive outcomes.


Bacteria in the lakes of the Tibetan Plateau and polar regions.
The Tibetan Plateau, also termed 'the Third Pole' harbors the largest number of high-altitude lakes in the world. Due to the presence of extreme conditions such as low temperature and oligotrophy, the lakes of the Tibetan Plateau share environmental features in common with lakes in the polar regions. However, the extent to which these environments are analogous, or indeed whether they harbor similar microbial communities or a high level of endemic species is poorly understood. Here we compared high-throughput 16S rRNA gene sequencing data from the lakes of the three different regions in order to characterize their taxonomic diversity, the community composition and biogeography. Our results showed despite the similarity in environmental conditions, the spatial distribution of the bacterial communities was distinct with only 3.1% of all operational taxonomic units (OTUs) being present in all three regions (although these OTUs did account for a considerable proportion of the total sequences, 36.4%). Sequences belonging to Burkholderiales and Actinomycetales dominated the shared OTUs across all three regions. Scale dependent distance decay patterns provided evidence of dispersal limitation. Climatic variables and dispersal limitation were apparently both important in controlling the spatial distribution of bacterial communities across regions. This work expands our understanding of the diversity and biogeography of lake bacterial communities across the Tibetan Plateau and provides insights into how they compare to those of the Antarctic and Arctic.


How preserved is episodic memory in behavioral variant frontotemporal dementia?
Studies have shown variable memory performance in patients with behavioral variant frontotemporal dementia (bvFTD). Our study investigated whether this variability is due to the admixture of patients with true bvFTD and phenocopy patients. We also sought to compare performance of patients with bvFTD and patients with Alzheimer disease (AD). We analyzed neuropsychological memory performance in patients with a clinical diagnosis of bvFTD divided into those who progressed (n = 50) and those who remained stable (n = 39), patients with AD (n = 64), and healthy controls (n = 64). Patients with progressive bvFTD were impaired on most memory tests to a similar level to that of patients with early AD. Findings from a subset of patients with progressive bvFTD with confirmed FTLD pathology (n = 10) corroborated these findings. By contrast, patients with phenocopy bvFTD performed significantly better than progressors and patients with AD. Logistic regression revealed that patients with bvFTD can be distinguished to a high degree (85%) on the immediate recall score of a word list learning test (Rey Auditory Verbal Learning Test). Our results provide evidence for an underlying memory deficit in "real" or progressive behavioral variant frontotemporal dementia (bvFTD) similar to Alzheimer disease, though the groups differ in orientation scores, with patients with bvFTD being intact. Exclusion solely based on impaired neuropsychological memory performance can potentially lead to an underdiagnosis of FTD.


Structure and interactions of NCAM modules 1 and 2, basic elements in neural cell adhesion.
The structure in solution of the second Ig-module fragment of residues 117-208 of NCAM has been determined. Like the first Ig-module of residues 20-116, it belongs to the I set of the immunogloblin superfamily. Module 1 and module 2 interact weakly, and the binding sites of this interaction have been identified. The two-module fragment NCAM(20-208) is a stable dimer. Removal of the charged residues in these sites in NCAM(20-208) abolishes the dimerization. Modeling the dimer of NCAM(20-208) to fit the interactions of these charges produces one coherent binding site for the formation of two antiparallel strands of the first two NCAM modules. This mode of binding could be a major element in trans-cellular interactions in neural cell adhesion.


Effect of oximetry on hospitalization in bronchiolitis: a randomized clinical trial.
Routine use of pulse oximetry has been associated with changes in bronchiolitis management and may have lowered the hospitalization threshold for patients with bronchiolitis. To examine if infants with bronchiolitis whose displayed oximetry measurements have been artificially elevated 3 percentage points above true values experience hospitalization rates at least 15% lower compared with infants with true values displayed. Randomized, double-blind, parallel-group trial conducted from 2008 to 2013 in a tertiary-care pediatric emergency department in Toronto, Ontario, Canada. Participants were 213 otherwise healthy infants aged 4 weeks to 12 months with mild to moderate bronchiolitis and true oxygen saturations of 88% or higher. Pulse oximetry measurements with true saturation values displayed or with altered saturation values displayed that have been increased 3 percentage points above true values. The primary outcome was hospitalization within 72 hours, defined as inpatient admission within this interval or active hospital care for greater than 6 hours. Secondary outcomes included the use of supplemental oxygen in the emergency department, level of physician agreement with discharge from the emergency department, length of emergency department stay, and unscheduled visits for bronchiolitis within 72 hours. Forty-four of 108 patients (41%) in the true oximetry group and 26 of 105 (25%) in the altered oximetry group were hospitalized within 72 hours (difference, 16% [95% CI for the difference, 3.6% to 28.4%]; P = .005). Using the emergency department physician as a random effect, the primary treatment effect remained significant (adjusted odds ratio, 4.0 [95% CI, 1.6 to 10.5]; P = .009). None of the secondary outcomes were significantly different between the groups. There were 23 of 108 (21.3%) subsequent unscheduled medical visits for bronchiolitis in the true oximetry group and 15 of 105 (14.3%) in the altered oximetry group (difference, 7% [95% CI, -0.3% to 0.2%]; P = .18). Among infants presenting to an emergency department with mild to moderate bronchiolitis, those with an artificially elevated pulse oximetry reading were less likely to be hospitalized within 72 hours or to receive active hospital care for more than 6 hours than those with unaltered oximetry readings. This suggests that oxygen saturation should not be the only factor in the decision to admit, and its use may need to be reevaluated. clinicaltrials.gov Identifier: NCT00673946.


Association between emergency department resources and diagnosis of intimate partner violence.
There is little information about which intimate partner violence (IPV) policies and services assist in the identification of IPV in the emergency department (ED). The objective of this study was to examine the association between a variety of resources and documented IPV diagnoses. Using billing data assembled from 21 Oregon EDs from 2001 to 2005, we identified patients who were assigned a discharge diagnosis of IPV. We then surveyed ED directors and nurse managers to gain information about IPV-related policies and services offered by participating hospitals. We combined billing data, survey results, and hospital-level variables. Multivariate analysis assessed the likelihood of receiving a diagnosis of IPV depending on the policies and services available. In 754 597 adult female ED visits, IPV was diagnosed 1929 times. Mandatory IPV screening and victim advocates were the most commonly available IPV resources. The diagnosis of IPV was independently associated with the use of a standardized intervention checklist (odds ratio: 1.71; 95% confidence interval: 1.04-2.82). Public displays regarding IPV were negatively associated with IPV diagnosis (odds ratio 0.56; 95% confidence interval: 0.35-0.88). IPV remains a rare documented diagnosis. Most common hospital-level resources did not demonstrate an association with IPV diagnoses; however, a standardized intervention checklist may play a role in clinician's likelihood of diagnosing IPV.


[Primary melanoma of the esophagus].
Primary malignant melanoma of the esophagus is a rare, aggressive and poor-prognostic neoplasm, usually detected in late stages. Surgery (esophagectomy) is the treatment of choice when operable and the only factor that improves the disease prognosis is the detection of the lesions in early stages and the recognition of non-typical lesions. Most commonly, Melanoma lesions appear as large masses with proliferative, ulcerated and pigmented aspect. We report a clinical case whose endoscopic presentation was an elevated, protruding and sessile lesion (Paris Classification Type 0- Is), amelanocytic, size of 4mm, smooth surface, pink and with regular borders (mucosa of tumor with a normal esophagic mucosa appearance). This lesion initially could not be identified because the patient was admitted due to upper gastrointestinal bleeding caused by peptic esophagitis Grade D according to Los Angeles Classification. After histological analysis and immunohistochemistry of the lesion, melanoma was diagnosed. No skin, mucous or ocular lesions were found and multislice spiral computed tomography of thorax and abdomen did not show any metastasis.


Machinability investigation in turning of high density fiberboard.
A series of experiments were conducted to assess the machinability of high density fiberboard using cemented carbide cutting tools. The objective of this work was to investigate the influence of two cutting parameters, spindle speed and feed per turn, on cutting forces, chip formation and cutting quality. The results are as follows: cutting forces and chip-breaking length decrease with increasing spindle speed and decreasing feed per turn. In contrast, surface roughness increases with decrease of spindle speed and increase in feed per turn. Chips were divided into four categories based on their shape: dust, particle, splinter, and semicontinuous chips. Chip-breaking length had a similar tendency to the variance of cutting forces with respect to average roughness and mean peak-to-valley height: an increase in the variance of cutting forces resulted in increased average roughness and mean peak-to-valley height. Thus, high cutting speed and low feed rate are parameters suitable for high-quality HDF processing and will improve not only machining quality, but production efficiency.


Exaggerated postural vasoconstrictor reflex in Raynaud's phenomenon.
The central and local regulation of capillary blood flow in the finger was studied by the local xenon-133 washout technique in women with primary Raynaud's phenomenon, men with vibration induced white finger, and their respective sex matched controls. The vasoconstrictor response to venous stasis of 40 mm Hg elicited by local reflex was normal in both types of Raynaud's phenomenon. Change in posture from lying to sitting induced vasoconstriction in all groups, which was abolished by proximal nervous blockade. The vasoconstrictor response to sitting was augmented in both groups of subjects with Raynaud's phenomenon compared with their sex matched controls. These results show the existence of central and local postural vasoconstrictor reflexes in normal fingers. In both types of Raynaud's phenomenon there was hyperreactivity of the central sympathetic nervous system to orthostatic stress and normal function of digital arterioles and postganglionic sympathetic nerve fibres.


TNFR1 is essential for CD40, but not for lipopolysaccharide-induced sickness behavior and clock gene dysregulation.
Autoimmune and infectious diseases are associated with behavioral changes referred to as sickness behavior syndrome (SBS). In autoimmunity, the generation of anti-self T lymphocytes and autoantibodies critically involves binding of CD40 ligand on T-cells to its receptor CD40 on B-cells, dendritic cells and macrophages. Activation of CD40 leads to production of proinflammatory cytokines and, as shown here, induces SBS. Here we report that these behavioral changes depend on the expression of tumor necrosis factor alpha receptor 1 (TNFR1), but not on interleukin-1 receptor 1 or interleukin-6. Moreover, the intensity of SBS correlates with suppression of E-box controlled clock genes, including Dbp, and upregulation of Bmal1. However, the absence of TNFR1 does not interfere with the development of SBS and dysregulation of clock genes in mice treated with lipopolysaccharide. Thus, our results suggest that TNFR1 mediates SBS and dysregulation of clock genes in autoimmune diseases.


A Matter of Perspective: Comparison of the Characteristics of Persons with HIV Infection in the United States from the HIV Outpatient Study, Medical Monitoring Project, and National HIV Surveillance System.
Comparative analyses of the characteristics of persons living with HIV infection (PLWH) in the United States (US) captured in surveillance and other observational databases are few. To explore potential joint data use to guide HIV treatment and prevention in the US, we examined three CDC-funded data sources in 2012: the HIV Outpatient Study (HOPS), a multisite longitudinal cohort; the Medical Monitoring Project (MMP), a probability sample of PLWH receiving medical care; and the National HIV Surveillance System (NHSS), a surveillance system of all PLWH. Overall, data from 1,697 HOPS, 4,901 MMP, and 865,102 NHSS PLWH were analyzed. Compared with the MMP population, HOPS participants were more likely to be older, non-Hispanic/Latino white, not using injection drugs, insured, diagnosed with HIV before 2009, prescribed antiretroviral therapy, and to have most recent CD4+ T-lymphocyte cell count ≥500 cells/mm3 and most recent viral load test<2 00 copies/mL. The MMP population was demographically similar to all PLWH in NHSS, except it tended to be slightly older, HIV diagnosed more recently, and to have AIDS. Our comparative results provide an essential first step for combined epidemiologic data analyses to inform HIV care and prevention for PLWH in the US.


Early evolutionary loss of the lipid A modifying enzyme PagP resulting in innate immune evasion in Yersinia pestis.
Immune evasion through membrane remodeling is a hallmark of Yersinia pestis pathogenesis. Yersinia remodels its membrane during its life cycle as it alternates between mammalian hosts (37 °C) and ambient (21 °C to 26 °C) temperatures of the arthropod transmission vector or external environment. This shift in growth temperature induces changes in number and length of acyl groups on the lipid A portion of lipopolysaccharide (LPS) for the enteric pathogens Yersinia pseudotuberculosis (Ypt) and Yersinia enterocolitica (Ye), as well as the causative agent of plague, Yersinia pestis (Yp). Addition of a C16 fatty acid (palmitate) to lipid A by the outer membrane acyltransferase enzyme PagP occurs in immunostimulatory Ypt and Ye strains, but not in immune-evasive Yp Analysis of Yp pagP gene sequences identified a single-nucleotide polymorphism that results in a premature stop in translation, yielding a truncated, nonfunctional enzyme. Upon repair of this polymorphism to the sequence present in Ypt and Ye, lipid A isolated from a Yp pagP+ strain synthesized two structures with the C16 fatty acids located in acyloxyacyl linkage at the 2' and 3' positions of the diglucosamine backbone. Structural modifications were confirmed by mass spectrometry and gas chromatography. With the genotypic restoration of PagP enzymatic activity in Yp, a significant increase in lipid A endotoxicity mediated through the MyD88 and TRIF/TRAM arms of the TLR4-signaling pathway was observed. Discovery and repair of an evolutionarily lost lipid A modifying enzyme provides evidence of lipid A as a crucial determinant in Yp infectivity, pathogenesis, and host innate immune evasion.


Absorbable collagen membranes for periodontal regeneration: a systematic review.
Guided tissue regeneration (GTR) with bioabsorbable collagen membranes (CM) is commonly used for the treatment of periodontal defects. The objective of this systematic review of randomized clinical trials was to assess the clinical efficacy of GTR procedures with CM, with or without bone substitutes, in periodontal infrabony defects compared with that of open flap debridement (OFD) alone. Primary outcomes were tooth loss and gain in clinical attachment level (CAL). Screening of records, data extraction, and risk-of-bias assessments were performed by two reviewers. Weighted mean differences were estimated by random effects meta-analysis. We included 21 reports on 17 trials. Risk of bias was generally high. No data were available for the primary outcome tooth loss. The summary treatment effect for change in CAL for GTR with CM compared with OFD was 1.58 mm (95% CI, 1.27 to 1.88). Despite large between-trial heterogeneity (I2 = 75%, p < .001), all trials favored GTR over OFD. No differences in treatment effects were detected between trials of GTR with CM alone and trials of GTR with CM in combination with bone substitutes (p for interaction, .31). GTR with CM, with or without substitutes, may result in improved clinical outcomes compared with those achieved with OFD alone. Our findings support GTR with CM for the treatment of infrabony periodontal defects.


Validity and reliability of functional performance tests in meniscectomized patients with or without knee osteoarthritis.
Reduced functional performance and muscular dysfunction after knee injury and in knee osteoarthritis (OA) is suggested to be a factor in OA development. Validated functional performance tests applicable in the clinic and large-scale studies are lacking. The aim was to study the reliability and validity of 10 functional performance tests. Two hundred and eighty-five subjects, 15-22 years post-meniscectomy, performed 10 functional performance tests. The mean age was 54 years (SD+/-11.2) and 79% were men; 52% had radiographic OA, and 48% were categorized as symptomatic. The tests were evaluated for test-retest reliability, discriminative ability (younger vs older age, men vs women, symptom-free vs symptomatic) and floor and ceiling effects. Two of the 10 tests, maximum number of knee bendings in 30 s and one-leg hop for distance, had good test-retest reliability (ICC 0.92, 95% CI 0.86-0.96 and 0.93, 95% CI 0.87-0.97) and were able to discriminate with regard to age, gender and symptoms, and had acceptable floor effects (9% and 3%, respectively). This study suggests the use of two functional performance tests: knee bendings/30 s and one-leg hop for distance, easy to use for evaluation of interventions due to knee injury and knee OA and when attaining long-term data of natural disease history.


Epidermal growth factor receptors in human breast carcinoma cells: a potential selective target for transforming growth factor alpha-Pseudomonas exotoxin 40 fusion protein.
Epidermal growth factor (EGF) receptors are expressed in high levels by some poor prognosis breast tumors. We have examined the cytotoxic effect of the tumor growth factor alpha (TGF alpha)-delta Cys-Pseudomonas exotoxin (PE40) recombinant fusion protein on normal and tumorigenic human breast epithelial cells in vitro and in vivo. The MDA-468, MDA-231, BT-20, and MCF-7ADR estrogen receptor-negative, EGF receptor-rich breast cancer lines were exquisitely sensitive in vitro to TGF alpha-delta Cys-PE40 with a 50% inhibitory concentration of < or = 0.02 nM. The estrogen receptor-positive, low EGF receptor MCF-7, ZR75-1, and T47D cells were less sensitive to the fusion toxin with a 50% inhibitory concentration of > 0.2 nM. The nontumorigenic cell lines 184, 184A1, and 184B5 were relatively resistant to TGF alpha-delta Cys-PE40 despite exhibiting high levels of EGF receptors. Continuous i.p. administration of TGF alpha-delta Cys-PE40 via an osmotic minipump at a dose of 0.4 microgram/g/day over 7 days inhibited MDA-468, MA-231, and BT-20 but not MCF-7 tumor growth in female athymic mice. Host tissue toxicity was not observed with this dose of TGF alpha-delta Cys-PE40. Mixed MDA-468/MCF-7 tumors were established in nude mice after coinoculation of both cell types in estrogen-supplemented animals. EGF receptor immunohistochemistry and immunoblot procedures indicated that TGF alpha-PE40 eliminated the MDA-468 cells while sparing the adjacent MCF-7 cells. By immunoblot, EGF receptors were consistently more abundant in tumor tissue than in adjacent nontumor tissue from the same mastectomy specimen (n = 7). These data support the notion that EGF receptors can be selectively targeted in human breast cancer cells for the delivery of antitumor agents. Further clinical studies with TGF alpha-delta Cys-PE40 and other chimeric toxins using the same cellular target will address this possibility.


Effects of specific mu and kappa opiate tolerance and abstinence on hypothalamo-pituitary-adrenal axis secretion in the rat.
Chronic administration of opiates to rats results in HPA axis tolerance and abstinence-induced hypersecretion. The effects of specific mu and kappa tolerance and withdrawal on the functional secretion of the HPA axis were evaluated in this study. Adult male rats were injected s.c. twice daily with saline, morphine or U50,488 for 5 days. Serum adrenocorticotrophic hormone (ACTH) or corticosterone (CS) were determined by radioimmunoassay as measures of HPA axis function. Tolerance to morphine (10 mg/kg) and U50,488 (1 mg/kg), but no cross-tolerance, was observed suggesting the development of mu- or kappa-specific tolerance, respectively. Tolerance does not occur at the pituitary or adrenal levels after these paradigms because ACTH and CS responses to exogenous corticotropin-releasing factor and ACTH, respectively, were not attenuated. CS secretion in response to novelty stress was not affected by either chronic opiate treatment, but the circadian variation of CS levels was slightly blunted after chronic morphine. In contrast, the elevation of CS secretion by quipazine (0.5 mg/kg) and physostigmine (0.1 mg/kg) was attenuated after chronic U50,488, but not morphine administration. Both spontaneous and antagonist-precipitated withdrawal from morphine, but not U50,488, resulted in elevation of CS levels. Low doses of morphine suppressed morphine abstinence-induced CS hypersecretion, whereas, U50,488 and clonidine had no effect. In conclusion, alterations of HPA axis function occur during chronic mu or kappa opiate administration that are receptor-specific and involve multiple neural controls of the HPA axis.


Student, Faculty, and Staff Approval of University Smoke/Tobacco-Free Policies: An Analysis of Campus Newspaper Articles.
To provide a nontraditional source of data to university policymakers regarding student, faculty, and staff approval of university smoke/tobacco-free policies, as published through campus newspaper articles. From January to April 2016, a total of 2523 articles were retrieved concerning campus smoking/tobacco at 4-year, public universities. Of these, 54 articles met the inclusion factors, which described 30 surveys about campus approval of tobacco-free policies and 24 surveys about smoke-free policies. In all, the surveys included more than 130 000 respondents. With the exception of 4 surveys, all reported that the most of the respondents approved a tobacco/smoke-free campus policy. Although the study had several limitations, the findings provide a synthesis from a nontraditional data source that is consistent with findings from the peer-reviewed literature, in which most of the students, faculty, and staff on university campuses approve of smoke/tobacco-free campus policies.


Qualitative features of esophageal fluorescence angiography and anastomotic outcomes in children.
Indocyanine green (ICG) is commonly used to assess perfusion, but quality defining features are lacking. We sought to establish qualitative features of esophageal ICG perfusion assessments, and develop an esophageal anastomotic scorecard to risk-stratify anastomotic outcomes. Single institution, retrospective analysis of children with an intraoperative ICG perfusion assessment of an esophageal anastomosis. Qualitative perfusion features were defined and a perfusion score developed. Associations between perfusion and clinical features with poor anastomotic outcomes (PAO, leak or refractory stricture) were evaluated with logistic and time-to-event analyses. Combining significant features, we developed and tested an esophageal anastomotic scorecard to stratify PAO risk. From 2019 to 2021, 53 children (median age 7.4 months) underwent 55 esophageal anastomoses. Median (IQR) follow-up was 14 (10-19.9) months; mean (SD) perfusion score was 13.2 (3.4). Fifteen (27.3%) anastomoses experienced a PAO and had significantly lower mean perfusion scores (11.3 (3.3) vs 14.0 (3.2), p = 0.007). Unique ICG perfusion features, severe tension, and primary or rescue traction-induced esophageal lengthening [Foker] procedures were significantly associated with PAO on both logistic and Cox regression. The scorecard (range 0-7) included any Foker (+2), severe tension (+1), no arborization on either segment (+1), suture line hypoperfusion >twice expected width (+2), and segmental or global areas of hypoperfusion (+1). A scorecard cut-off >3 yielded a sensitivity of 73% and specificity of 93% (AUC 0.878 [95%CI 0.777 to 0.978]) in identifying a PAO. A scoring system comprised of qualitative ICG perfusion features, tissue quality, and anastomotic tension can help risk-stratify esophageal anastomotic outcomes accurately. Diagnostic - II.


A nearly cryptic Scorpionfly, Panorpa cryptica n. sp. (Mecoptera: Panorpidae) from North America.
The first nearly cryptic species of scorpionfly from the United States, Panorpa cryptica Bicha and Schiff, n. sp., is described from northern Georgia, southwestern North Carolina and northwestern South Carolina. This insect was initially differentiated from the very similar Panorpa nebulosa Westwood by its unique cytochrome oxidase subunit I (COI) mitochondrial DNA. Habitat details, distribution, and biology are described.


[Hereditary colorectal cancer].
Hereditary syndromes cause approximately 5 to 15% of overall colorectal cancer (CRC) cases. Hereditary CRC is conventionally divided into two major categories: hereditary non-polyposis colorectal cancer (HNPCC) and those related to polyposis syndromes including familial adenomatous polyposis (FAP), Peutz-Jegher syndrome (PJS), and juvenile polyposis (JP). The screening for the cancer and methods of treatment applied to patients with hereditary CRC are quite different from those applied to the general population. The genes responsible for these syndromes has recently identified, as a result, genetic testing has become the most important determining factor in clinical decisions. Germ-line mutation of the APC gene induces FAP, an autosomal dominant disorder, characterized by the development of hundreds to thousands of colonic adenomas. CRC appears in almost all affected individuals by the time they are 50 years of age. An affected individual should undergo colectomy by his/her late teens. Furthermore, according to the findings of genetic testing, at-risk family members also need endoscopic surveillance and surgery. Recently, a mutation on the MYH gene is increasingly being investigated in patients with multiple polyps, and autosomal recessive MYH polyposis is considered to be a new category of polyposis. More common than FAP, HNPCC is caused by germ-line mutations in DNA mismatch repair genes, mainly MLH1 and MSH2. Although there is no polyposis, polyps seem to be more villous and dysplastic and appear to grow rapidly into CRCs. The aggregate lifetime risk of CRC is about 80% for mutation carriers. The risk for other types of cancer, such as endometrial, ovarian, small bowel, and transitional cell cancer, is also increased. The Amsterdam criteria and Bethesda guidelines are the best-known tools for diagnosis and genetic testing, and colectomy followed by endoscopic follow-up is the standard treatment. PJS and JP are reported to be characterized by hamartomatous polyps throughout the GI tract and germ-line mutations in the STK11 gene (PJS) and the DPC4/BMPR1A gene (JP).


Epitope specificity of anti-synapsin autoantibodies: Differential targeting of synapsin I domains.
To identify the specific domains of the presynaptic protein synapsin targeted by recently described autoantibodies to synapsin. Sera of 20 and CSF of two patients with different psychiatric and neurological disorders previously tested positive for immunoglobulin (Ig)G antibodies to full-length synapsin were screened for IgG against synapsin I domains using HEK293 cells transfected with constructs encoding different domains of rat synapsin Ia. Additionally, IgG subclasses were determined using full-length synapsin Ia. Serum and CSF from one patient were also screened for IgA autoantibodies to synapsin I domains. Sera from nine and CSF from two healthy subjects were analyzed as controls. IgG in serum from 12 of 20 IgG synapsin full-length positive patients, but from none of the healthy controls, bound to synapsin domains. Of these 12 sera, six bound to the A domain, five to the D domain, and one to the B- (and possibly A-), D-, and E-domains of synapsin I. IgG antibodies to the D-domain were also detected in one of the CSF samples. Determination of IgG subclasses detected IgG1 in two sera and one CSF, IgG2 in none of the samples, IgG3 in two sera, and IgG4 in eight sera. One patient known to be positive for IgA antibodies to full-length synapsin had IgA antibodies to the D-domain in serum and CSF. Anti-synapsin autoantibodies preferentially bind to either the A- or the D-domain of synapsin I.


Are nitrogen-fertilized forest soils sinks or sources of carbon?
We developed a simple conceptual model that tracks nitrogen and carbon jointly through an N fertilized forest ecosystem. The stimulation of growth increases the litterfall and imports substrate for soil microorganisms. Microbial biomass forms according to the supply of C and N. The formation of microbial biomass is accompanied by respiratory C losses. The quantity of CO2 efflux depends on the C use efficiency of microbes. When excess N is available, the microbial activity is accelerated and the demand for substrate is high. Litterfall supplies an insufficient amount of C to the soil. In such a case, labile soil C is mineralized and the net effect of N fertilization is a loss of soil C. A strong N fertilization effect on the aboveground biomass can offset the soil C loss. In the case of a low N dosage or high N losses due to leaching or emission of nitrogen oxides, the soil C loss is small. The conceptual model was applied to a case study. The field data, collected over a time span of several decades, could not support sound conclusions on the temporal trend of soil C because the spatial and temporal variability of the chemical data was high. The conceptual model allowed to give an evaluation of the fertilization effect on soil C based on reproducible principles.


The amplification model for adaptive mutation: simulations and analysis.
It has been proposed that the lac revertants arising under selective conditions in the Cairns experiment do not arise by stress-induced mutagenesis of stationary phase cells as has been previously assumed. Instead, these revertants may arise within growing clones initiated by cells with a preexisting duplication of the weakly functional lac allele used in this experiment. It is proposed that spontaneous stepwise increases in lac copy number (amplification) allow a progressive improvement in growth. Reversion is made more likely primarily by the resultant increase in the number of mutational targets--more cells with more lac copies. The gene amplification model requires no stress-induced variation in the rate or target specificity of mutation and thus does not violate neo-Darwinian theory. However, it does require that a multistep process of amplification, reversion, and amplification segregation be completed within approximately 20 generations of growth. This work examines the proposed amplification model from a theoretical point of view, formalizing it into a mathematical framework and using this to determine what would be required for the process to occur within the specified period. The analysis assumes no stress-induced change in mutation rate and describes only the growth improvement occurring during the process of amplification and subsequent elimination of excess mutant lac copies. The dynamics of the system are described using Monte Carlo simulations and numerical integration of the deterministic equations governing the system. The results imply that the amplification model can account for the behavior of the system using biologically reasonable parameter values and thus can, in principle, explain Cairnsian adaptive mutation.


Guard Cells Integrate Light and Temperature Signals to Control Stomatal Aperture.
High temperature promotes guard cell expansion, which opens stomatal pores to facilitate leaf cooling. How the high-temperature signal is perceived and transmitted to regulate stomatal aperture is, however, unknown. Here, we used a reverse-genetics approach to understand high temperature-mediated stomatal opening in Arabidopsis (Arabidopsis thaliana). Our findings reveal that high temperature-induced guard cell movement requires components involved in blue light-mediated stomatal opening, suggesting cross talk between light and temperature signaling pathways. The molecular players involved include phototropin photoreceptors, plasma membrane H+-ATPases, and multiple members of the 14-3-3 protein family. We further show that phototropin-deficient mutants display impaired rosette evapotranspiration and leaf cooling at high temperatures. Blocking the interaction of 14-3-3 proteins with their client proteins severely impairs high temperature-induced stomatal opening but has no effect on the induction of heat-sensitive guard cell transcripts, supporting the existence of an additional intracellular high-temperature response pathway in plants.


Resection of selected invasive squamous cell carcinoma of the pyriform sinus by means of the lateral pharyngotomy approach: the partial lateral pharyngectomy.
Although the lateral pharyngotomy is a well-known surgical procedure, to our knowledge, no published reports have described the results achieved with a lateral pharyngectomy approach in patients with isolated and previously untreated selected invasive squamous cell carcinoma of the lateral wall of the pyriform sinus. The medical files and operative charts of 30 patients with an isolated, and previously untreated, squamous cell carcinoma of the pyriform sinus, treated for cure by lateral partial pharyngectomy and primary closure, as well as postoperative radiotherapy (n = 22), and preoperative chemotherapy (n = 8), were retrospectively studied. Sixteen tumors were classified as T1 and 14 as T2. All patients but one were followed until death. Three patients (9%) died in the immediate postoperative period (two from medical complications and one from a surgery-related complication). Overall, 29 patients died, resulting in a 77.7%, 40%, 23.3%, and 15% 1-, 3-, 5- and 10-year Kaplan-Meier actuarial survival estimate, respectively. Only one patient had a serious postoperative complication develop. All patients were decannulated. No patients required a gastrostomy and/or completion total laryngectomy for functional reasons. Local recurrence occurred in four patients (13%). The 3-and 5-year Kaplan-Meier actuarial local control estimates were 88.5% and 79.6%, respectively. The use of a platin-based induction chemotherapy (p = .05) regimen was the only variable that was significantly statistically related to local recurrence. Overall, a 93.3% laryngeal preservation rate was achieved. Partial pharyngectomy by means of the lateral pharyngotomy combined with postoperative radiation therapy is an efficient and function-sparing approach to control selected invasive squamous cell carcinomas of the lateral wall of the pyriform sinus.


EGFR gene copy number increase in vulvar carcinomas is linked with poor clinical outcome.
EGFR copy number increases have been frequently reported in cancer including vulvar carcinomas. Co-amplification of cancer genes plays an important role in the development of many tumour types. To better understand the effect of EGFR aberrations on vulvar cancer phenotype and patient prognosis, the authors analysed EGFR copy number changes using fluorescence in situ hybridisation and EGFR expression by immunohistochemistry in a tissue microarray containing 183 squamous cell carcinomas of vulva. Furthermore, the authors analysed the co-amplification frequency of EGFR with HER2, CCND1, MYC and PIK3CA, respectively. EGFR copy number increase was found in 39.3% of the tumours. Seventeen per cent of vulvar carcinomas showed EGFR high polysomy including 9% with amplification of the EGFR gene. Copy number gain of the EGFR locus was associated with non-basaloid phenotype (p=0.03), high-tumour stage (p<0.001), human papillomaviruse negativity of tumours (p=0.04) and the number of lymph node metastases (p=0.02). EGFR protein expression was statistically correlated to EGFR copy number increase (p<0.05). The observed co-amplification rate of EGFR with all four additionally examined oncogenes was much higher than statistically expected. There was a highly significant association between EGFR copy number increase and CCND1 amplifications (p<0.001) as well as the total number of gene amplifications (p=0.04). EGFR copy number gains were significantly related to unfavourable patient outcome in univariate analysis and multivariate Cox regression analysis. In conclusion, EGFR copy number increases are detectable in a substantial proportion of vulvar carcinomas with relationships to advanced tumour stages and the development of lymph node metastases. EGFR copy number aberrations are connected to other gene amplifications and probably define an human papillomaviruses-independent pathway in the development of vulvar carcinomas. These data support the potential utility of EGFR inhibitors as a therapeutic alternative in a subset of vulvar carcinomas.


Sticking and patching: tuning and anchoring cyclometallated ruthenium(II) complexes.
A series of [Ru(bpy)2(C^N)][PF6] (HC^N = 2-phenylpyridine derivative) complexes functionalized in the cyclometallating C^N phenyl ring with F, Me, OMe, CO2Me, S(t)Bu, SO2Me (ligands H1-H6) or in the C^N pyridine ring with 4-CO2Me or 4-C6H4P(O)(OEt)2 substituents (ligands H7 or H9) have been prepared and characterized; representative crystal structures confirm the structural features of the complexes. When the C^N ligand contains a CO2H substituent (ligand H28), deprotonation in addition to cyclometallation occurs to give a neutral, zwitter-ionic complex [Ru(bpy)2(8)]. The synthesis of the cationic complexes requires addition of a silver(i) salt (AgPF6 or AgBF4) to abstract Cl(-) from cis-[Ru(bpy)2Cl2] and (1)H NMR spectroscopic data are consistent with interactions between Ag(+) and the coordinated C^N ligand in [Ru(bpy)2(C^N)](+). The absorption spectra of [Ru(bpy)2(C^N)][PF6] (C^N = 1-6) are similar, but the introduction of the anchoring domains in [Ru(bpy)2(C^N)][PF6] with C^N = 7 or 9 enhances the absorption response; the greatest influence is observed in [Ru(bpy)2(9)](+) with the introduction of the 4-C6H4P(O)(OEt)2 substituent. Trends in emission and electrochemical behaviours of the complexes are interpreted in terms of the influence of the electronic properties of the C^N ligand substituents on the energies of the HOMO which is localized on the C^N ligand and Ru centre. This study provides an optimized synthetic route to the phosphonate ester derivative [Ru(bpy)2(9)][PF6], designed to anchor to a semiconductor surface; this complex also exhibits the most favourable absorption properties among the complexes studied.


Concurrent and predictive validity of a self-reported measure of medication adherence.
Adherence to the medical regimen continues to rank as a major clinical problem in the management of patients with essential hypertension, as in other conditions treated with drugs and life-style modification. This article reviews the psychometric properties and tests the concurrent and predictive validity of a structured four-item self-reported adherence measure (alpha reliability = 0.61), which can be easily integrated into the medical visit. Items in the scale address barriers to medication-taking and permit the health care provider to reinforce positive adherence behaviors. Data on patient adherence to the medical regimen were collected at the end of a formalized 18-month educational program. Blood pressure measurements were recorded throughout a 3-year follow-up period. Results showed the scale to demonstrate both concurrent and predictive validity with regard to blood pressure control at 2 years and 5 years, respectively. Seventy-five percent of the patients who scored high on the four-item scale at year 2 had their blood pressure under adequate control at year 5, compared with 47% under control at year 5 for those patients scoring low (P less than 0.01).