A nitrogen response pathway regulates virulence functions in Fusarium oxysporum via the protein kinase TOR and the bZIP protein MeaB.
During infection, fungal pathogens activate virulence mechanisms, such as host adhesion, penetration and invasive growth. In the vascular wilt fungus Fusarium oxysporum, the mitogen-activated protein kinase Fmk1 is required for plant infection and controls processes such as cellophane penetration, vegetative hyphal fusion, or root adhesion. Here, we show that these virulence-related functions are repressed by the preferred nitrogen source ammonium and restored by treatment with l-methionine sulfoximine or rapamycin, two specific inhibitors of Gln synthetase and the protein kinase TOR, respectively. Deletion of the bZIP protein MeaB also resulted in nitrogen source-independent activation of virulence mechanisms. Activation of these functions did not require the global nitrogen regulator AreA, suggesting that MeaB-mediated repression of virulence functions does not act through inhibition of AreA. Tomato plants (Solanum lycopersicum) supplied with ammonium rather than nitrate showed a significant reduction in vascular wilt symptoms when infected with the wild type but not with the DeltameaB strain. Nitrogen source also affected invasive growth in the rice blast fungus Magnaporthe oryzae and the wheat head blight pathogen Fusarium graminearum. We propose that a conserved nitrogen-responsive pathway might operate via TOR and MeaB to control virulence in plant pathogenic fungi.


IL-17 Production by γδ+ T Cells Is Critical for Inducing Th17 Responses in the Female Genital Tract and Regulated by Estradiol and Microbiota.
IL-17 can be produced by adaptive immune cells such as Th17 cells and by immune cells that produce IL-17 without prior priming. This latter category, which we will refer to as "innate," includes innate cells such as NK cells and innate lymphoid cells and innate-like T cell populations such as NKT cells and γδ+ T cells. Studies in mucosal tissues have shown that the induction of Th17 immunity is amplified by innate IL-17 produced within those tissues. However, the role of innate IL-17 and its effect on Th17 induction in the female genital tract (FGT) is largely unknown. In this study, we characterize the primary source of IL-17-secreting vaginal cells and show that innate IL-17 plays a critical role in priming adaptive Th17 responses in the FGT. Under homeostatic conditions, γδ+ T cells were the predominant source of innate IL-17 in the murine FGT, and this population was modulated by both the sex hormone estradiol and the presence of commensal microbiota. Compared with wild-type C57BL/6 mice, vaginal APCs isolated from IL-17A-deficient (IL-17A-/- ) mice were severely impaired at priming Th17 responses in APC-T cell cocultures. Furthermore, the defect in Th17 induction in the absence of innate IL-17 was associated with impairment of IL-1β production by vaginal CD11c+ dendritic cells. Overall, our study describes a novel role for IL-17 in the FGT and further demonstrates the importance of factors in the vaginal microenvironment that can influence adaptive immune responses.


Asthma, allergy, and atopy in three south-east Asian populations.
Whilst many recent reports have suggested a rise in the prevalence of asthma and allergic disease in Western countries, little is known about the epidemiology of these common conditions in south-east Asia. This study compared the prevalence of asthma and allergic disease amongst secondary school students in three south-east Asian populations--Hong Kong, Kota Kinabalu in Malaysia, and San Bu in China--and investigated the associations with atopy and family history. Secondary school students were given standard questionnaires on respiratory and allergic symptoms for completion by parents with response rates of 89.2% in Hong Kong (611 male, 451 female; mean (SD) age = 13.9 (1.8 years), 87.6% in Kota Kinabalu (134 male, 275 female; 15.5 (2.1) years), and 98.6% in San Bu (492 male, 245 female; 16.4 (1.8) years). Skin tests were performed in a subsample of students to determine atopic status. The respective prevalence (and 95% CI) for hayfever, eczema, and wheeze or asthma were 15.7% (13.5, 17.9), 20.1% (17.7, 22.5), 11.6% (9.3, 13.9) in Hong Kong, 11.2% (8.2, 14.3), 7.6% (5.0, 10.1), 8.2% (5.5, 10.9) in Kota Kinabalu, and 2.1% (1.2, 3.1), 7.2% (5.4, 9.1), 1.9% (0.7, 3.1) in San Bu. Atopy was common and was present in 49.0-63.9% of subjects in the three populations. Dust mite and cockroach were the commonest allergens that gave positive reactions in 42.8-60.5% and 25.7-35.9% of students respectively. A higher proportion of students in Hong Kong had severe degree of reactivity on skin test than the other two populations. Family history was associated with asthma and allergic symptoms in the three populations conferring a 3-80-fold increase in risk to family members and was a stronger predictor for asthma and allergy than atopy. Prevalence of asthma and allergic disease is low compared with Western countries, but considerable differences exist between the three south-east Asian populations despite similar rates of atopy. Asthma and allergic disease are more strongly associated with family history than atopy, which suggests that genetic and environmental factors common to the family, other than aeroallergen sensitisation, are important in the pathogenesis of asthma and allergy in the region.


High-dose methylprednisolone in the treatment of active ulcerative colitis.
Twenty patients with severe active ulcerative colitis were entered into a pilot study to test the efficacy of pulsed high-dose methylprednisolone in inducing remission. Of 20 patients, 12 improved, but 8 of 20 patients (40%) who did not went for urgent colectomy. The group proceeding to surgery tended to have more extensive disease. These results contrast unfavorably with historical control data from this unit.


In-Field Validation of an Inertial Sensor-Based System for Movement Analysis and Classification in Ski Mountaineering.
Ski Mountaineering (SkiMo) is a fast growing sport requiring both endurance and technical skills. It involves different types of locomotion with and without the skis. The aim of this study is to develop and validate in the snowfield a novel inertial-based system for analysing cycle parameters and classifying movement in SkiMo in real-time. The study was divided into two parts, one focused on real-time parameters estimation (cadence, distance from strides, stride duration, stride length, number of strides, slope gradient, and power) and, second, on transition detection (kickturns, skin on, skin off, ski on and off backpack) in order to classify between the different types of locomotion. Experimental protocol involved 16 experienced subjects who performed different SkiMo trials with their own equipment instrumented with a ski-mounted inertial sensor. The results obtained by the algorithm showed precise results with a relative error near 5% on all parameters. The developed system can, therefore, be used by skiers to obtain quantitative training data analysis and real-time feedback in the field. Nevertheless, a deeper validation of this algorithm might be necessary in order to confirm the accuracy on a wider population of subjects with various skill levels.


Circum-menarcheal bone acquisition is stress-driven: A longitudinal study in adolescent female gymnasts and non-gymnasts.
Mechanical loading through youth exercise is highly modifiable and represents a strategy to maximize peak adult bone mass, with the potential for broad implementation across the population to lower fracture risk. For girls, circum-menarcheal growth is critical, with around 50% of adult bone acquired over a 4-year period. Here, we prospectively followed 10 gymnasts and 12 age-matched non-gymnasts across approximately 4 years circum-menarche. A combination of pQCT and subject-specific finite element models were used to measure differences in bone acquisition and structure between the groups, and to determine the degree to which specific mechanical factors predict change in bone structure. At baseline, gymnasts had stronger bone, including 26% higher BMC, 51% greater compressive strength, and 21% higher trabecular density. Over the study period, both groups more than doubled their bone strength. Pre-menarcheal principal stresses predicted change in pQCT variables for non-gymnasts, but not gymnasts. The bone of non-gymnasts became more asymmetrical than the bone of gymnasts. Our results suggest that exposure to the diverse, intense mechanical signals of gymnastic loading during adolescence imparts substantial benefits to bone geometry and mechanical function. Specifically, the bone of gymnasts is better able to resist loading from multiple directions, and operates with a higher factor of safety compared to non-gymnasts.


Improved information pooling for hierarchical cognitive models through multiple and covaried regression.
Cognitive process models are fit to observed data to infer how experimental manipulations modify the assumed underlying cognitive process. They are alternatives to descriptive models, which only capture differences on the observed data level, and do not make assumptions about the underlying cognitive process. Process models may require more observations than descriptive models however, and as a consequence, usually fewer conditions can be simultaneously modeled with them. Unfortunately, it is known that the predictive validity of a model may be compromised when fewer experimental conditions are jointly accounted for (e.g., overestimation of predictor effects, or their incorrect assignment). We develop a hierarchical and covaried multiple regression approach to address this problem. Specifically, we show how to map the recurrences of all conditions, participants, items, and/or traits across experimental design cells to the process model parameters. This systematic pooling of information can facilitate parameter estimation. The proposed approach is particularly relevant for multi-factor experimental designs, and for mixture models that parameterize per cell to assess predictor effects. This hierarchical framework provides the capacity to model more conditions jointly to improve parameter recovery at low observation numbers (e.g., using only 1/6 of trials, recovering as well as standard hierarchical Bayesian methods), and to directly model predictor and covariate effects on the process parameters, without the need for post hoc analyses (e.g., ANOVA). An example application to real data is also provided.


Resolution improvement in STED super-resolution microscopy at low power using a phasor plot approach.
Stimulated emission depletion (STED) microscopy is a powerful super-resolution microscopy technique that has achieved significant results in breaking the resolution limit and relevant applications. In principle, STED super resolution is obtained by stimulated emission partially inhibiting the spontaneous emission in the periphery of a diffraction-limited area. However, very high depletion laser power is generally necessary for the enhancement of imaging resolution, which is harmful to live biological specimens due to its high phototoxicity and photo-bleaching effects. Therefore, further improving the STED resolution at a lower depletion power level has recently attracted increasing interest from researchers in various fields. In this work, a phasor plot approach combined with fluorescence lifetime imaging microscopy (FLIM) is used to resolve the abovementioned problem based on a long- and short-lifetime criterion. Firstly, the time-resolved data obtained by STED-FLIM is converted to the frequency domain via a phasor approach. Next, partial data is extracted according to the information on the phase and amplitude for resolution improvement. Then, fluorescent microspheres (100 nm in diameter) are observed under different depletion powers, resulting in a series of improved resolution through phasor plots. Finally, this method is applied to image human Nup153 in fixed HeLa cells, providing a 86 nm higher resolution than that in traditional STED imaging at a depletion power of 20 mW.


The COVID-19 crisis does not diminish environmental motivation: Evidence from two panel studies of decision making and self-reported pro-environmental behavior.
The literature shows that threats unrelated to environmental problems can shift attention away from these problems and affect pro-environmental behavior. It is not clear whether the COVID-19 crisis that started in 2019 had any uniform effect on pro-environmental behavior and decision making. In two preregistered panel studies conducted before and during the first COVID wave (n1 = 206, n2 = 164) and before and during the second COVID wave (n3 = n4 = 260), we found that the crisis had had no uniform effect on pro-environmental behaviors, environmental attitude, nor on the behavioral costs of general pro-environmental behavior. Analysis of one specific pro-environmental behavior, the choice of environmentally friendly delivery of products, revealed that the general preference for green delivery services and heightened preference for green delivery services among people with higher attitude levels remained unchanged by the COVID-19 crisis. Thus, if the COVID-19 crisis has had any effects on pro-environmental behaviors, these effects are probably fragmented, specific to certain population segments, and not visible in the short-term perspective.


Risk factors for a first-incident radiographic vertebral fracture in women > or = 65 years of age: the study of osteoporotic fractures.
Vertebral fractures in older women signal an increased risk of additional osteoporotic fractures. To identify risk factors for first vertebral fractures, we studied 5822 women > or =65 years of age who had no fracture on baseline radiographs of the spine. Several modifiable risk factors increased an older woman's risk of developing a first vertebral fracture, and women with multiple risk factors and low BMD had the highest risk. Risk factors and low BMD should be useful to help focus efforts to prevent these fractures. Vertebral fractures are a common cause of back pain and disability and signal an increased risk of additional osteoporotic fractures in older women. Little is known about the risk factors for the first occurrence of a vertebral fracture. To identify risk factors for a first vertebral fracture, we studied 5822 women > or =65 years of age from the Study of Osteoporotic Fractures who had no fracture on baseline radiographs of the spine. We measured potential risk factors and BMD of the wrist and calcaneus at baseline and BMD of the spine and hip halfway through follow-up. Fractures were assessed by standard methods from spine radiographs obtained at baseline and follow-up an average of 3.7 years later. In multivariable analyses, older age, previous nonspine fracture, low BMD at all sites, a low body mass index (BMI), current smoking, low milk consumption during pregnancy, low levels of daily physical activity, having a fall, and regular use of aluminum-containing antacids independently increased the risk of a first vertebral fracture. Women using estrogen and those who engaged in recreational physical activity had a decreased risk. The effects of low BMI, smoking, use of estrogen and antacids, and previous fracture were partially mediated by BMD. Women in the lower third of wrist BMD with five or more risk factors had a 12-fold greater risk than women in the highest third of BMD who had zero to three risk factors. The 27% of women at highest risk suffered 60% of the incident fractures. In conclusion, several modifiable risk factors and BMD independently increase an older woman's risk of developing a first vertebral fracture. The combination of risk factors and BMD should be useful for focusing efforts to prevent vertebral fractures.


An alternative approach to the traditional mixotrophic cultures of Haematococcus pluvialis Flotow (Chlorophyceae).
In traditional mixotrophic cultures of microalgae, all the inorganic nutrients and organic carbon sources are supplied in the medium before inoculation. In this study, however, an alternative approach was adopted in Haematococcus pluvialis Flotow, a microalga capable of growing mixotrophically on sodium acetate (Na-Ac). First, the cells were grown under 75 micromol photons m-2 s-1 phototrophically without Na-Ac until the stationary phase and then exposed to five different light regimes by the addition of Na-Ac, e.g., dark, 20, 40, 75 and 150 micromol photons m-2 s-1. Dry weight (DW), pigments and especially cell number in alternative mixotrophy (AM) were higher than traditional mixotrophy (TM). Cell number in AM almost doubled up from 21.7 to 42.9 x 104 cells mL-1 during 5 day exposure to Na-Ac, whereas the increase was only 1.2-fold in TM. Maximum cell density was reached in 75 micromol photons m-2 s-1 among the light intensities tested. We propose that Na-Ac in TM of H. pluvialis can not be utilized as efficient as in AM. With this respect, AM is of several advantages against TM such as a much higher cell density in a batch culture period and minimized risk of contamination due to the shorter exposure of cells to organic carbon sources. In consequence, this method may be used for other strains of the species, and even for the other microalgal species able to grow mixotrophically.


The elusive gene for keratolytic winter erythema.
Keratolytic winter erythema (KWE), also known as Oudtshoorn skin disease, is characterised by a cyclical disruption of normal epidermal keratinisation affecting primarily the palmoplantar skin with peeling of the palms and soles, which is worse in the winter. It is a rare monogenic, autosomal dominant condition of unknown cause. However, due to a founder effect, it occurs at a prevalence of 1/7 200 among South African Afrikaans-speakers. In the mid-1980s, samples were collected from affected families for a linkage study to pinpoint the location of the KWE gene. A genome-wide linkage analysis, using microsatellite markers, identified the KWE critical region on chromosome 8p23.1-p22. Subsequent genetic studies focused on screening candidate genes in this critical region; however, no pathogenic mutations that segregated exclusively with KWE were identified. The cathepsin B (CTSB) and farnesyl-diphosphate farnesyltransferase 1 (FDFT1) genes revealed no potentially pathogenic variants, nor did they show differential gene expression in affected skin. Mutation detection in additional candidate genes also failed to identify the KWE-associated variant, suggesting that the causal variant may be in an uncharacterised functional region. Bioinformatic analysis revealed highly conserved regions within the KWE critical region and a custom tiling array was designed to cover this region and to search for copy number variation. Although the study did not identify a variant that segregates exclusively with KWE, it provided valuable insight into the complex KWE-linked region. Next-generation sequencing approaches are being used to comb the region, but the causal variant for this interesting hyperkeratotic palmoplantar phenotype still remains elusive.


The Use of Virtual Reality for Managing Psychological Distress in Adolescents: Systematic Review.
Acute and chronic psychological distress are prevalent during adolescence and can have negative impacts on adolescents in all life domains. The aim of this systematic review was to appraise the use of virtual reality (VR) interventions to manage symptoms of psychological distress symptoms among adolescents. MEDLINE, PubMed, PsycINFO, and Scopus databases were searched up to June 2020. Available citations were de-duplicated and screened by two authors using title and abstract information. A total of 301 articles were retained for full-text evaluation next to eligibility criteria. Empirical studies of all designs and comparator groups were included if these appraised the impact of an immersive VR intervention on any standardized measure indicative of psychological distress in an adolescent sample. Data were extracted into a standardized coding sheet. Results were tabulated and discussed with a narrative synthesis due to the heterogeneity between studies. A total of seven studies met inclusion criteria. There were four randomized controlled trials and three uncontrolled pilot studies on new VR interventions. Distress-related issues included: state-anxiety, venepuncture, risk taking, public speaking anxiety, social anxiety disorder, sexual victimization, and chemotherapy administration. All studies reported significant changes on outcome measures after VR treatment. Six studies reported small-to-large reductions in symptoms. The average attrition rate was 3.6 percent during the active VR treatment phase. Treatment acceptability was high in the studies that assessed user engagement factors. The VR technology can provide a safe, rapidly efficacious, and acceptable treatment modality for managing psychological distress in several key adolescent populations.


The Perception of Visual UncertaintyRepresentation by Non-Experts.
We tested how non-experts judge point probability for seven different visual representations of uncertainty, using a case from an unfamiliar domain. Participants (n = 140) rated the probability that the boundary between two earth layers passed through a given point, for seven different visualizations of the positional uncertainty of the boundary. For all types of visualizations, most observers appear to construct an internal model of the uncertainty distribution that closely resembles a normal distribution. However, the visual form of the uncertainty range (i.e., the visualization type) affects this internal model and the internal model relates to participants' numeracy. We conclude that perceived certainty is affected by its visual representation. In a follow-up experiment we found no indications that the absence (or presence) of a prominent center line in the visualization affects the internal model. We discuss if and how our results inform which visual representation is most suitable for representing uncertainty and make suggestions for future work.


Using intrinsically fluorescent proteins for plant cell imaging.
The intrinsically fluorescent proteins (IFPs), such as the green, cyan and yellow fluorescent proteins, have revolutionized how we can image the dynamics of cellular events. Intrinsically fluorescent proteins have been used as reporter genes to monitor transcriptional regulation, as targeted markers for organelles and subcellular structures, in fusion proteins to directly observe protein motility and dynamics, and in sensors designed to show changes in cellular environments ranging from pH to protein kinase activity. The IFPs hold tremendous potential to reveal the dynamic processes that underlie plant cell function; however, as with all technology there are artifacts and pitfalls inherent in their use. In this review, we highlight some of the practical issues in using IFPs for live cell imaging. These include choice of the appropriate IFP, dealing with autofluorescence, photobleaching and phototoxicity, and application of approaches such as fluorescence resonance energy transfer (FRET), fluorescence lifetime imaging (FLIM) and fluorescence recovery after photobleaching (FRAP) to gain high-resolution data about protein dynamics within the cell. We also discuss some of the more common artifacts associated with these fluorescence imaging approaches and suggest controls that should help both spot these problems and suggest their solutions.


Biogenesis, transit, and functional properties of the insulin proreceptor and modified insulin receptors in 3T3-L1 adipocytes. Use of monensin to probe proreceptor cleavage and generate altered receptor subunits.
The biogenesis, intracellular transport, and functional properties of the insulin proreceptor and modified insulin receptors were studied in hormone-responsive 3T3-L1 adipocytes. After control cells were labeled with [35S]Met for 7 min, the principal polypeptide that was precipitated by anti-insulin receptor antibodies had a molecular weight (Mr) of 180 000. This initial precursor was rapidly converted (t1/2 = 35 min) to a 200-kilodalton (kDa) polypeptide, designated the insulin proreceptor, by the apparent posttranslational addition of N-linked, high mannose core oligosaccharide units. Mature alpha (Mr 130 000) and beta (Mr 90 000) subunits were derived from sequences within the proreceptor by proteolytic cleavage and late processing steps, and these subunits appeared on the cell surface 2-3 h after synthesis of the 180-kDa precursor. The cation ionophore monensin was used in combination with metabolic labeling, affinity cross-linking, and external proteolysis to probe aspects of proreceptor function, transit, and the development of insulin sensitivity at the target cell surface. At 5 micrograms/mL, monensin potently inhibited the proteolytic cleavage step, and the 200-kDa polypeptide accumulated. Lower concentrations of the ionophore selectively blocked late processing steps in 3T3-L1 adipocytes so that apparently smaller alpha' (Mr 120 000) and beta' (Mr 85 000) subunits were produced. Proreceptor and alpha' and beta' subunits were translocated to the cell surface, indicating that the signal for intracellular transit occurs in the 200-kDa polypeptide and is independent of the posttranslational proteolysis and late processing steps. The alpha' subunit bound insulin both at the surface of intact cells and after solubilization with Triton X-100; the beta' subunit was phosphorylated in an insulin-stimulated manner. The detergent-solubilized 200-kDa proreceptor also exhibited both functional properties. However, the proreceptor that was transported to and exposed on the cell surface was incapable of binding insulin in intact adipocytes. Thus, late processing is not essential for the expression of functions associated with mature alpha and beta subunits. In contrast, it appears that the proteolytic generation of subunits is required for the correct orientation of the hormone binding site in the plasma membrane bilayer and the development of insulin responsiveness in 3T3-L1 adipocytes.


Sec24-dependent secretion drives cell-autonomous expansion of tracheal tubes in Drosophila.
Epithelial tubes in developing organs, such as mammalian lungs and insect tracheae, need to expand their initially narrow lumina to attain their final, functional dimensions. Despite its critical role for organ function, the cellular mechanism of tube expansion remains unclear. Tracheal tube expansion in Drosophila involves apical secretion and deposition of a luminal matrix, but the mechanistic role of secretion and the nature of forces involved in the process were not previously clear. Here we address the roles of cell-intrinsic and extrinsic processes in tracheal tube expansion. We identify mutations in the sec24 gene stenosis, encoding a cargo-binding subunit of the COPII complex. Via genetic-mosaic analyses, we show that stenosis-dependent secretion drives tube expansion in a cell-autonomous fashion. Strikingly, single cells autonomously adjust both tube diameter and length by implementing a sequence of events including apical membrane growth, cell flattening, and taenidial cuticle formation. Known luminal components are not required for this process. Thus, a cell-intrinsic program, rather than nonautonomous extrinsic cues, controls the dimensions of tracheal tubes. These results indicate a critical role of membrane-associated proteins in the process and imply a mechanism that coordinates autonomous behaviors of individual cells within epithelial structures.


Super-Nyquist-WDM transmission over 7,326-km seven-core fiber with capacity-distance product of 1.03 Exabit/s · km.
We show super-Nyquist-WDM transmission technique, where optical signals with duobinary-pulse shaping can be wavelength-multiplexed with frequency spacing of below baudrate. Duobinary-pulse shaping can reduce the signal bandwidth to be a half of baudrate while controlling inter-symbol interference can be compensated by the maximum likelihood sequence estimation in a receiver. First, we experimentally evaluate crosstalk characteristics as a function of channel spacing between the dual-channel DP-QPSK signals with duobinary-pulse shaping. As a result, the crosstalk penalty can be almost negligible as far as the ratio of baudrate to frequency spacing is maintained to be less than 1.20. Next, we demonstrate 140.7-Tbit/s, 7,326-km transmission of 7 × 201-channel 25-GHz-spaced super-Nyquist-WDM 100-Gbit/s optical signals using seven-core fiber and full C-band seven-core EDFAs. To the best of our knowledge, this is one of the first reports of high-capacity transmission experiments with capacity-distance product in excess of 1 Exabit/s · km.


Summary of the IADR Cariology Research Group Symposium, Barcelona, Spain, July 2010: new directions in cariology research.
Caries remains the most prevalent noncontagious biofilm-mediated disease in humans. It is clear that the current approaches to decrease the prevalence of caries in human populations, including water fluoridation and school-based programs, are not enough to protect everyone. The scientific community has suggested the need for innovative work in a number of areas in cariology, encompassing disease etiology, epidemiology, definition, prevention, and treatment. In this symposium, two of these areas, dealing specifically with etiological aspects of caries were discussed: (1) systematic research on caries risk assessment using population-based cohort techniques, and (2) genetic studies to identify genes and genetic markers of diagnostic, prognostic, and therapeutic value. This paper summarizes these presentations.


Lexical-semantic organization in children with specific language impairment.
To determine whether children with specific language impairment (SLI) show deficits in lexical-semantic organization and, if so, whether these deficits are commensurate with their delay in vocabulary size and whether the deficits affect all children with SLI. Fourteen children with SLI, 14 age matches (AM), and 14 expressive vocabulary matches (VM) generated 3 associations to each of 48 words. Associations were coded as semantic (e.g., dog-pet), clang (e.g., cow-how), or erroneous (e.g., spoon-Disney). Relative to the AM children, children with SLI produced fewer semantic responses, more clangs, and more errors. Relative to the VM children, fewer semantic responses and more errors in the children with SLI were found in by-item analyses. Across elicitation trials, semantic responses decreased in the AM and VM children but remained stable in the SLI children. Examination of individual performance in the SLI group revealed that poor semantic performance was associated with a deficit in expressive vocabulary and a gap between receptive and expressive vocabularies. Significant variability in lexical-semantic organization skills exists among children with SLI. Deficits in lexical-semantic organization were demonstrated by a subgroup of children with SLI who likely had concomitant word-finding difficulties.


Simulated consultations: a sociolinguistic perspective.
Assessment of consulting skills using simulated patients is widespread in medical education. Most research into such assessment is sited in a statistical paradigm that focuses on psychometric properties or replicability of such tests. Equally important, but less researched, is the question of how far consultations with simulated patients reflect real clinical encounters--for which sociolinguistics, defined as the study of language in its socio-cultural context, provides a helpful analytic lens. In this debate article, we draw on a detailed empirical study of assessed role-plays, involving sociolinguistic analysis of talk in OSCE interactions. We consider critically the evidence for the simulated consultation (a) as a proxy for the real; (b) as performance; (c) as a context for assessing talk; and (d) as potentially disadvantaging candidates trained overseas. Talk is always a performance in context, especially in professional situations (such as the consultation) and institutional ones (the assessment of professional skills and competence). Candidates who can handle the social and linguistic complexities of the artificial context of assessed role-plays score highly--yet what is being assessed is not real professional communication, but the ability to voice a credible appearance of such communication. Fidelity may not be the primary objective of simulation for medical training, where it enables the practising of skills. However the linguistic problems and differences that arise from interacting in artificial settings are of considerable importance in assessment, where we must be sure that the exam construct adequately embodies the skills expected for real-life practice. The reproducibility of assessed simulations should not be confused with their validity. Sociolinguistic analysis of simulations in various professional contexts has identified evidence for the gap between real interactions and assessed role-plays. The contextual conditions of the simulated consultation both expect and reward a particular interactional style. Whilst simulation undoubtedly has a place in formative learning for professional communication, the simulated consultation may distort assessment of professional communication These sociolinguistic findings contribute to the on-going critique of simulations in high-stakes assessments and indicate that further research, which steps outside psychometric approaches, is necessary.


Expression of the type 1 insulin-like growth factor receptor is up-regulated in primary prostate cancer and commonly persists in metastatic disease.
The type 1 insulin-like growth factor receptor (IGF1R) mediates tumor cell growth, adhesion, and protection from apoptosis. High plasma IGF-I levels predispose to prostate cancer, but there is no consensus regarding IGF1R expression in primary and metastatic prostate cancer. Recent studies in a human cell line and a mouse model suggest that metastatic prostate cancer cell detachment may be favored by impairing cadherin function via loss of expression of insulin receptor substrate-1 (IRS-1), the principal IGF1R docking molecule. This may be accompanied by PTEN mutation, reactivating a key antiapoptotic pathway, and by IGF1R down-regulation to prevent Shc-mediated differentiation. We studied IGF1R expression in 54 samples of primary prostate tissue including 44 archival and 10 prospectively collected biopsies. We performed semiquantitative immunostaining for the IGF1R, IRS-1, and PTEN, and in situ hybridization for IGF1R. The IGF1R was significantly up-regulated at the protein and mRNA level in primary prostate cancer compared with benign prostatic epithelium. There was a trend toward increased expression of IRS-1 in the malignant biopsies. We also measured IGF1R, IRS-1, and PTEN expression in 12 paired biopsies of primary prostate cancer and subsequent bone metastases. In four cases, IGF1R and IRS-1 levels were lower in the metastases than in the primary tumors. Three of these metastases also lacked significant PTEN staining, compatible with findings in the model systems described above. However, this pattern was relatively uncommon, and 8 of 12 cases expressed detectable IGF1R and IRS-1 in both primary and metastatic biopsies. These findings challenge earlier reports of IGF1R down-regulation in metastatic disease and reinforce the importance of the IGF1R in prostate cancer biology.


Dynamic simulation of articulated rigid bodies with contact and collision.
We propose a novel approach for dynamically simulating articulated rigid bodies undergoing frequent and unpredictable contact and collision. In order to leverage existing algorithms for nonconvex bodies, multiple collisions, large contact groups, stacking, etc., we use maximal rather than generalized coordinates and take an impulse-based approach that allows us to treat articulation, contact, and collision in a unified manner. Traditional constraint handling methods are subject to drift, and we propose a novel prestabilization method that does not require tunable potentially stiff parameters as does Baumgarte stabilization. This differs from poststabilization in that we compute allowable trajectories before moving the rigid bodies to their new positions, instead of correcting them after the fact when it can be difficult to incorporate the effects of contact and collision. A poststabilization technique is used for momentum and angular momentum. Our approach works with any black box method for specifying valid joint constraints and no special considerations are required for arbitrary closed loops or branching. Moreover, our implementation is linear both in the number of bodies and in the number of auxiliary contact and collision constraints, unlike many other methods that are linear in the number of bodies, but not in the number of auxiliary constraints.


A Scoping Review on COVID-19 Vaccine Hesitancy among the Lesbian, Gay, Bisexual, Transgender, Queer, Intersex and Asexual (LGBTQIA+) Community and Factors Fostering Its Refusal.
The ramifications of the existing crisis caused by the coronavirus pandemic are sensed in all walks of life. Among the various efforts made to curb the spread of this novel infection, the development of COVID-19 vaccines had a profound role in flattening the pandemic curve. Even though the rapid vaccine drive received a highly welcoming response among people, the reluctance and ignorance of a part of the population towards available safe vaccines stand as impediments to achieving the desired outcome. The LGBTQIA+ (Lesbian, Gay, Bisexual, Transgender, Queer, Intersex and Asexual) communities are the least studied groups in this regard. The purpose of this study is to extensively review and report on COVID-19 vaccine uptake and refusal among the LGBTQIA+ population and enumerate the factors contributing to vaccine hesitancy. The study extends further to outline a conceptual framework for interventions to enhance COVID-19 vaccine acceptance among the LGBTQIA+ population. We performed a systematic search using key terms on Google Scholar and PubMed. The obtained results were filtered using the eligibility criteria framed for this study. The initial search provided an extensive result of 4510 articles which were later screened at various levels to arrive at the final inclusive collection of manuscripts adding to 17. The studies were analyzed by the authors individually, and the data were categorized using variables. The results are interpreted using charts and graphs. The whole manuscript has been structured in accordance with the PRISMA extension for scoping reviews. The comprehensive search yielded 17 eligible articles for this review. Most of the studies were conducted in the United States (n = 17), and predominantly cross-sectional studies have been conducted. The major comparative factor was the HIV status of the LGBTQIA+ population. HIV-affected patients were more willing to take up COVID-19 vaccination. However, social stigma, discrimination, lack of access and non-prioritization in vaccine drives were found to be the major factors contributing to vaccine hesitancy among this population. The invention of the COVID-19 vaccination revolutionized the healthcare systems burdened with COVID-19. Although this is a breakthrough scientific contribution, many factors are associated with the rate of vaccine acceptance, especially among sexual and gender minorities. The reviewed studies have revealed numerous factors that influence vaccine uptake and refusal with the commonest being concerns on discrimination, social stigma, inequitable access to healthcare, vaccine safety, efficacy, potency, side effects and lack of trust in medical workers. These impediments in vaccine coverage should be meticulously addressed to ensure optimum LGBTQIA+ physical and mental health as well as for providing non-discriminative, equitable and quality healthcare service regardless of the gender or sexual orientation of individuals.


A porcine model of neurofibromatosis type 1 that mimics the human disease.
Loss of the NF1 tumor suppressor gene causes the autosomal dominant condition, neurofibromatosis type 1 (NF1). Children and adults with NF1 suffer from pathologies including benign and malignant tumors to cognitive deficits, seizures, growth abnormalities, and peripheral neuropathies. NF1 encodes neurofibromin, a Ras-GTPase activating protein, and NF1 mutations result in hyperactivated Ras signaling in patients. Existing NF1 mutant mice mimic individual aspects of NF1, but none comprehensively models the disease. We describe a potentially novel Yucatan miniswine model bearing a heterozygotic mutation in NF1 (exon 42 deletion) orthologous to a mutation found in NF1 patients. NF1+/ex42del miniswine phenocopy the wide range of manifestations seen in NF1 patients, including café au lait spots, neurofibromas, axillary freckling, and neurological defects in learning and memory. Molecular analyses verified reduced neurofibromin expression in swine NF1+/ex42del fibroblasts, as well as hyperactivation of Ras, as measured by increased expression of its downstream effectors, phosphorylated ERK1/2, SIAH, and the checkpoint regulators p53 and p21. Consistent with altered pain signaling in NF1, dysregulation of calcium and sodium channels was observed in dorsal root ganglia expressing mutant NF1. Thus, these NF1+/ex42del miniswine recapitulate the disease and provide a unique, much-needed tool to advance the study and treatment of NF1.


The impact of official development aid on maternal and reproductive health outcomes: a systematic review.
Progress toward meeting Millennium Development Goal 5, which aims to improve maternal and reproductive health outcomes, is behind schedule. This is despite ever increasing volumes of official development aid targeting the goal, calling into question the distribution and efficacy of aid. The 2005 Paris Declaration on Aid Effectiveness represented a global commitment to reform aid practices in order to improve development outcomes, encouraging a shift toward collaborative aid arrangements which support the national plans of aid recipient countries (and discouraging unaligned donor projects). We conducted a systematic review to summarise the evidence of the impact on MDG 5 outcomes of official development aid delivered in line with Paris aid effectiveness principles and to compare this with the impact of aid in general on MDG 5 outcomes. Searches of electronic databases identified 30 studies reporting aid-funded interventions designed to improve maternal and reproductive health outcomes. Aid interventions appear to be associated with small improvements in the MDG indicators, although it is not clear whether changes are happening because of the manner in which aid is delivered. The data do not allow for a meaningful comparison between Paris style and general aid. The review identified discernible gaps in the evidence base on aid interventions targeting MDG 5, notably on indicators MDG 5.4 (adolescent birth rate) and 5.6 (unmet need for family planning). This review presents the first systematic review of the impact of official development aid delivered according to the Paris principles and aid delivered outside this framework on MDG 5 outcomes. Its findings point to major gaps in the evidence base and should be used to inform new approaches and methodologies aimed at measuring the impact of official development aid.


Young women partition fatty acids towards ketone body production rather than VLDL-TAG synthesis, compared with young men.
Before the menopause, women are relatively protected against CVD compared with men. The reasons for this sex difference are not completely understood, but hepatic fatty acid metabolism may play a role. The present study aimed to investigate the utilisation of plasma NEFA by the liver and to determine whether they are partitioned differently into ketone bodies and VLDL-TAG in healthy, lean young men and women. Volunteers were studied during a prolonged overnight fast (12-19 h) using an intravenous infusion of [U-¹³C]palmitate. After 12 h fasting, the women had a more advantageous metabolic profile with lower plasma glucose (P < 0·05) and TAG (P < 0·05) but higher plasma NEFA (P < 0·05) concentrations. Plasma 3-hydroxybutyrate (3-OHB) concentrations rose more in women than in men, and the transfer of ¹³C from [U-¹³C]palmitate to plasma [¹³C]3-OHB reached a plateau 6-7 h after the start of the infusion in women but was still increasing at 6 h in men. This implies a slower 3-OHB production rate and/or dilution by other precursor pools in men. In women, the high isotopic enrichment of plasma 3-OHB suggested that systemic plasma fatty acids were the major source of 3-OHB production. However, in men, this was not observed during the course of the study (P < 0·01). There were no sex differences for the incorporation of ¹³C into VLDL1- or VLDL2-TAG. The ability of young women to partition fatty acids towards ketone body production rather than VLDL-TAG may contribute to their more advantageous metabolic profile compared with young men.


Contrasting effects of acute and chronic stress on the transcriptome, epigenome, and immune response of Atlantic salmon.
Stress experienced during early life may have lasting effects on the immune system, with impacts on health and disease dependent on the nature and duration of the stressor. The epigenome is especially sensitive to environmental stimuli during early life and represents a potential mechanism through which stress may cause long-lasting health effects. However, the extent to which the epigenome responds differently to chronic vs acute stressors is unclear, especially for non-mammalian species. We examined the effects of acute stress (cold-shock during embryogenesis) and chronic stress (absence of tank enrichment during larval-stage) on global gene expression (using RNA-seq) and DNA methylation (using RRBS) in the gills of Atlantic salmon (Salmo salar) four months after hatching. Chronic stress induced pronounced transcriptional differences, while acute stress caused few lasting transcriptional effects. However, both acute and chronic stress caused lasting and contrasting changes in the methylome. Crucially, we found that acute stress enhanced transcriptional immune response to a pathogenic challenge (bacterial lipopolysaccharide, LPS), while chronic stress suppressed it. We identified stress-induced changes in promoter and gene-body methylation that were associated with altered expression for a small proportion of immune-related genes, and evidence of wider epigenetic regulation within signalling pathways involved in immune response. Our results suggest that stress can affect immuno-competence through epigenetic mechanisms, and highlight the markedly different effects of chronic larval and acute embryonic stress. This knowledge could be used to harness the stimulatory effects of acute stress on immunity, paving the way for improved stress and disease management through epigenetic conditioning.


Accurate method to determine the eigenstates of polarization in gyrotropic media.
Many practical modulator materials include combinations of electrooptically induced birefringence, optical activity, and/or Faraday rotation. Thus, there is a need for a procedure to design and analyze devices fabricated with materials exhibiting any or all of these effects. In this paper a simple procedure employing an extension of the general Jacobi method is introduced for determining the properties of the two allowed elliptical eigenpolarizations for an arbitrary direction of propagation and for the principal indices and axes of a general lossless, electrooptic, and gyrotropic medium. The procedure uses an iterative application of unitary transformations to diagonalize the Hermitian impermeability tensor. A complex polarization variable is defined from elements of the unitary transformation matrix to determine the ellipticity, azimuth angle, relative amplitude and phase, and handedness of the two orthogonal elliptical polarizations. The phase velocity indices of refraction are readily calculated with simple derived expressions. The procedure is numerically stable and accurate for any crystal class, external field direction, and direction of propagation.


Effects of local alendronate administration on bone defect healing. Histomorphometric and radiological evaluation in a rabbit model.
To performed a histomorphometric and radiological study to evaluate the effects of alendronate sodium administered locally in mandibular bone defects created in rabbits. Two circular defects 5 mm in diameter were created bilaterally in the mandibular corpus of 20 New Zealand rabbits (i.e., four defects per animal). Each defect received one of four treatments: no treatment (EC group), alendronate irrigation (AL group), autogenous bone grafting (AG group), or alendronate irrigation with autogenous bone grafting (AL+AG group). Histomorphometric and radiological assessments were conducted at 4 and 8 weeks after surgery. Between-group comparisons of the new bone area, the value of the AL+AG group was significantly lower thanthe remaining three groups at 4 weeks postoperatively. In all groups, the new bone area was significantly larger at 8 weeks than at 4 weeks. The residual graft area at 4 and 8 weeks was significantly higher in the AL+AG group than in the AG group, although it was significantly smaller at 8 weeks than at 4 weeks in both these groups. The use of alendronate sodium in conjunction with autogenous bone grafting improves the osteoconductive properties of the graft, enhances graft retention in the defect, and improves ossification.


First Report of Boscalid and Penthiopyrad-Resistant Isolates of Alternaria solani Causing Early Blight of Potato in Michigan.
Early blight of potato (Solanum tuberosum) is caused by Alternaria solani and occurs annually in Michigan. If left uncontrolled, it can result in yield losses exceeding 20% and impact stored potatoes. The disease is commonly managed using succinate dehydrogenase inhibitor (SDHI) fungicides (1). Unfortunately, recent studies have shown that SDHI resistance has increased dramatically over the past 2 years in A. solani populations (1,2). To investigate the occurrence of SDHI resistance in Michigan, potato leaves with early blight symptoms were collected from fields in Montcalm and Ionia counties, MI, in 2012. To obtain A. solani isolates from leaves, small pieces of leaf tissue (5 × 5 mm) were excised from the center of lesions and transferred on to water agar. Plates were incubated at 25°C overnight to allow conidia to germinate. Single germinated A. solani conidia were transferred to potato dextrose agar (PDA) and incubated at 25°C for 7 days. The identity of cultures was confirmed by colony and conidial morphology (3). Nineteen A. solani isolates were obtained and each was screened for sensitivity to the SDHI fungicides boscalid, penthiopyrad, and fluopyram, using a 50 ppm discriminatory dose based on EC50 values previously determined (2). Mycelial plugs (~5.5 mm) were transferred to amended and non-amended PDA plates that were incubated at 25°C for 7 days. An isolate was considered highly resistant if fungal growth relative to control plates exceeded 50%, moderately resistant if it was between 35 and 50%, and sensitive if it was less than 35% (2). A sensitive A. solani isolate (AS11) from Bonners Ferry, ID, was used as a control in these experiments. Of all isolates tested, 11% were highly resistant to both boscalid and penthiopyrad and 5% were moderately resistant to both fungicides, 21% were moderately resistant to penthiopyrad alone, and the remaining isolates (84 and 68% respectively) were sensitive to the two fungicides. None of the isolates tested were resistant to fluopyram. Recently, two major mutations, H227R in SdhB and H133R in SdhD, have been identified in highly resistant A. solani isolates in Idaho (2). Because the majority of the identified mutations occur near the 3' end of each subunit, this region was amplified and sequenced using the following primer sets: SdhB (5'-TACTGGTGGAACCAGGAGGAGTA-3' and 5'-CATACCACTCTAGGTGAAG-3'), SdhC (5'-CCAAATCACCTGGTACGCCTCG-3' and 5'-TCATCCGAGGAAGGTGTAGTAAAGGCTG-3'), and SdhD (5'-CCGACTCTATTCTCTGCGCCCT-3' and 5'-CTCGAAAGAGTAGAGGGCAAGACCCA-3'). In this study, all of the isolates that were highly resistant to both boscalid and penthiopyrad were found to contain the H133R mutation in SdhD, which correlated with the strongest resistance phenotype. To our knowledge, this is the first report of resistance to SDHI fungicides in populations of A. solani on potato in Michigan. These data are concerning as they indicate that the highly resistant isolates have already developed cross-resistance between boscalid and penthiopyrad, despite penthiopyrad not yet having regular use in Michigan. Although all of the isolates tested were sensitive to fluopyram, the discovery of isolates resistant to boscalid and penthiopyrad suggests that all SDHI fungicides should be considered at high risk of resistance development in Michigan. References: (1) K. Fairchild et al. Crop Prot. 49:31, 2013. (2) T. Miles et al. Plant Pathol. doi: 10.1111/ppa.12077, 2013. (3) P. Wharton et al. Plant Dis. 96:454, 2012.


Contribution of an unusual CDR2 element of a single domain antibody in ricin toxin binding affinity and neutralizing activity.
Ricin toxin's enzymatic subunit (RTA) has been subjected to intensive B cell epitope mapping studies using a combination of competition ELISAs, hydrogen exchange-mass spectrometry and X-ray crystallography. Those studies identified four spatially distinct clusters (I-IV) of toxin-neutralizing epitopes on the surface of RTA. Here we describe A9, a new single domain camelid antibody (VHH) that was proposed to recognize a novel epitope on RTA that straddles clusters I and III. The X-ray crystal structure of A9 bound to RTA (2.6 Å resolution) revealed extensive antibody contact with RTA's β-strand h (732 Å2 buried surface area; BSA), along with limited engagement with α-helix D (90 Å2) and α-helix C (138 Å2). Collectively, these contacts explain the overlap between epitope clusters I and III, as identified by competition ELISA. However, considerable binding affinity, and, consequently, toxin-neutralizing activity of A9 is mediated by an unusual CDR2 containing five consecutive Gly residues that interact with α-helix B (82 Å2), a known neutralizing hotspot on RTA. Removal of a single Gly residue from the penta-glycine stretch in CDR2 reduced A9's binding affinity by 10-fold and eliminated toxin-neutralizing activity. Computational modeling indicates that removal of a Gly from CDR2 does not perturb contact with RTA per se, but results in the loss of an intramolecular hydrogen bond network involved in stabilizing CDR2 in the unbound state. These results reveal a novel configuration of a CDR2 element involved in neutralizing ricin toxin.


Broad spectrum developmental role of Brachypodium AUX1.
Targeted cellular auxin distribution is required for morphogenesis and adaptive responses of plant organs. In Arabidopsis thaliana (Arabidopsis), this involves the prototypical auxin influx facilitator AUX1 and its LIKE-AUX1 (LAX) homologs, which act partially redundantly in various developmental processes. Interestingly, AUX1 and its homologs are not strictly essential for the Arabidopsis life cycle. Indeed, aux1 lax1 lax2 lax3 quadruple knock-outs are mostly viable and fertile, and strong phenotypes are only observed at low penetrance. Here we investigated the Brachypodium distachyon (Brachypodium) AUX1 homolog BdAUX1 by genetic, cell biological and physiological analyses. We report that BdAUX1 is essential for Brachypodium development. Bdaux1 loss-of-function mutants are dwarfs with aberrant flower development, and consequently infertile. Moreover, they display a counter-intuitive root phenotype. Although Bdaux1 roots are agravitropic as expected, in contrast to Arabidopsis aux1 mutants they are dramatically longer than wild type roots because of exaggerated cell elongation. Interestingly, this correlates with higher free auxin content in Bdaux1 roots. Consistently, their cell wall characteristics and transcriptome signature largely phenocopy other Brachypodium mutants with increased root auxin content. Our results imply fundamentally different wiring of auxin transport in Brachypodium roots and reveal an essential role of BdAUX1 in a broad spectrum of developmental processes, suggesting a central role for AUX1 in pooideae.


Late HIV Testing in a Cohort of HIV-Infected Patients in Puerto Rico.
Late HIV testing (LT), defined as receiving an AIDS diagnosis within a year of one's first positive HIV test, is associated with higher HIV transmission, lower HAART effectiveness, and worse outcomes. Latinos represent 36% of LT in the US, yet research concerning LT among HIV cases in Puerto Rico is scarce. Multivariable logistic regression analysis was used to identify factors associated with LT, and a Cochran‒Armitage test was used to determine LT trends in an HIV-infected cohort followed at a clinic in Puerto Rico specialized in the management and treatment of HIV. From 2000 to 2011, 47% of eligible patients were late testers, with lower median CD4 counts (54 vs. 420 cells/mm3) and higher median HIV viral load counts (253,680 vs. 23,700 copies/mL) than non-LT patients. LT prevalence decreased significantly, from 47% in 2000 to 37% in 2011. In a mutually adjusted logistic regression model, males, older age at enrollment and past history of IDU significantly increased LT odds, whereas having a history of amphetamine use decreased LT odds. When the data were stratified by mode of transmission, it became apparent that only the category men who have sex with men (MSM) saw a significant reduction in the proportion of LT, falling from 67% in 2000 to 33% in 2011. These results suggest a gap in early HIV detection in Puerto Rico, a gap that decreased only among MSM. An evaluation of the manner in which current HIV-testing guidelines are implemented on the island is needed.


Endotoxin removal by magnetic separation-based blood purification.
This work describes a magnetic separation-based approach using polymyxin B-functionalized metal alloy nanomagnets for the rapid elimination of endotoxins from human blood in vitro and functional assays to evaluate the biological relevance of the blood purification process. Playing a central role in gram-negative sepsis, bacteria-derived endotoxins are attractive therapeutic targets. However, both direct endotoxin detection in and removal from protein-rich fluids remains challenging. We present the synthesis and functionalization of ultra-magnetic cobalt/iron alloy nanoparticles and a magnetic separation-based approach using polymyxin B-functionalized nanomagnets to remove endotoxin from human blood in vitro. Conventional chromogenic Limulus Amebocyte Lysate assays confirm decreased endotoxin activity in purified compared to untreated samples. Functional assays assessing key steps in host defense against bacteria show an attenuated inflammatory mediator expression from human primary endothelial cells in response to purified blood samples compared to untreated blood and less chemotactic activity. Exposing Escherichia coli-positive blood samples to polymyxin B-functionalized nanomagnets even impairs the ability of gram-negative bacteria to form colony forming units, thus making magnetic separation based blood purification a promising new approach for future sepsis treatment.


Studying host-pathogen interactions in 3-D: organotypic models for infectious disease and drug development.
Representative, reproducible, and high-throughput models of human cells and tissues are critical for a meaningful evaluation of host-pathogen interactions and are an essential component of the research developmental pipeline. The most informative infection models-animals, organ explants, and human trials-are not suited for extensive evaluation of pathogenesis mechanisms and screening of candidate drugs. At the other extreme, more cost-effective and accessible infection models such as conventional cell culture and static coculture may not capture physiological and three-dimensional (3-D) aspects of tissue biology that are important in assessing pathogenesis, effectiveness, and cytotoxicity of therapeutics. Our lab has used innovative bioengineering technology to establish biologically meaningful 3-D models of human tissues that recapitulate many aspects of the differentiated structure and function of the parental tissue in vivo, and we have applied these models to study infectious disease. We have established a variety of different 3-D models that are currently being used in infection studies-including small intestine, colon, lung, placenta, bladder, periodontal ligament, and neuronal models. Published work from our lab has shown that our 3-D models respond to infection with bacterial and viral pathogens in ways that reflect the infection process in vivo. By virtue of their physiological relevance, 3-D cell cultures may also hold significant potential as models to provide insight into the neuropathogenesis of HIV infection. Furthermore, the experimental flexibility, reproducibility, cost efficiency, and high-throughput platform afforded by these 3-D models may have important implications for the design and development of drugs with which to effectively treat neurological complications of HIV infection.


Ultrasensitive, real-time analysis of biomarkers in breath using tunable external cavity laser and off-axis cavity-enhanced absorption spectroscopy.
A robust biomedical sensor for ultrasensitive detection of biomarkers in breath based on a tunable external cavity laser (ECL) and an off-axis cavity-enhanced absorption spectroscopy (OA-CEAS) using an amplitude stabilizer is developed. A single-mode, narrow-linewidth, tunable ECL is demonstrated. A broadly coarse wavelength tuning range of 720 cm⁻¹ for the spectral range between 6890 and 6170 cm⁻¹ is achieved by rotating the diffraction grating forming a Littrow-type external-cavity configuration. A mode-hop-free tuning range of 1.85 cm⁻¹ is obtained. The linewidths below 140 kHz are recorded. The ECL is combined with an OA-CEAS to perform laser chemical sensing. Our system is able to detect any molecule in breath at concentrations to the ppbv range that have absorption lines in the spectral range between 1450 and 1620 nm. Ammonia is selected as target molecule to evaluate the performance of the sensor. Using the absorption line of ammonia at 6528.76 cm⁻¹, a minimum detectable absorption coefficient of approximately 1×10⁻⁸ cm⁻¹ is demonstrated for 256 averages. This is achieved for a 1.4-km absorption path length and a 2-s data-acquisition time. These results yield a detection sensitivity of approximately 8.6×10⁻¹⁰ cm⁻¹ Hz(-1/2). Ammonia in exhaled breath is analyzed and found in a concentration of 870 ppb for our example.


Long-term effects of wildfire on available soil nutrient composition and stoichiometry in a Chinese boreal forest.
Wildfire disturbance is a major driver of biogeochemical processes in Eurasian boreal forests, yet little is known about the response of soil nutrient stoichiometry to wildfire in this ecosystem. To fill this gap, we measured the composition of available soil nutrients and their stoichiometric ratios in a Chinese boreal forest along a gradient of fire history. In the 1-year-post-fire site, wildfire increased the relative abundances of element nitrogen (N), phosphorus (P), Sulphur (S), iron (Fe), and aluminum (Al), and reduced the abundances of element calcium (Ca), magnesium (Mg), and potassium (K). Available soil N:K, N:S, P:K, P:S, and S:K ratios were 240%, 70%, 440%, 160%, and 150% higher than the control, but N:P ratio was not significantly different. In the 11-year-post-fire site, the soil nutrient composition recovered to the pre-fire levels. Although most of the soil nutrient stoichiometry returned to pre-fire levels, soil N:P ratios became significantly higher. These results showed immediate wildfire effects on soil nutrient availability and composition were strongly related to fire severity, but such effects could be subdued by soil environment and topographical variations over time. Although wildfire effects on soil nutrients are mostly short-term, it could produce relatively long-term effects on balance between N and P.


Replicase-derived resistance against pea early browning virus in Nicotiana benthamiana is an unstable resistance based upon posttranscriptional gene silencing.
Virus resistance in Nicotiana benthamiana plants containing a translatable Pea early browning virus (PEBV) 54K sequence from the 201K replicase gene has been reported previously. Resistant plants contain multiple transgene copies divided between two loci. Analysis of a genetic series containing the two loci in separate homozygous or heterozygous condition suggest that only one of the loci is necessary to induce the resistance. The resistance observed in R2 and R3 generations of lines containing both transgene loci in homozygous condition became less consistent in R4 and R5 generations. This inversely correlated with steady-state transgene transcript levels of the segregating populations. The use of recombinant Potato virus X vectors carrying PEBV 54K sequences showed that the resistance is based upon posttranscriptional gene silencing, is non-strand specific, and recognizes 3' located sequences within the PEBV 54K sequence.


[Comparison of Stages of Physical Activity Among Nurses, Physicians, and Allied Healthcare Professionals].
Despite the fact that regular physical activity is known to improve physical health and reduce absenteeism and perceived job strain, healthcare professionals currently perform inadequate physical activity. To understand and compare the differences in stages of physical activity among nurses, physicians, and allied healthcare professionals. A cross-sectional questionnaire was used to survey nurses, physicians, and allied healthcare professionals who worked at four branches of a hospital in Taiwan. Data were collected using the physical activity stage of change questionnire. Data were analyzed using descriptive analysis, Chi-square test of homogeneity of proportions and a posteriori comaparisons, and multinominal logistic regression analysis. Of the 3,260 questionnaires administrated, 2,001 valid questionnaires were returned for an effective response rate of 61.4%. In total, 1,497 (74.8%) of the nurses, 241 (12.0%) of the physicians, and 263 (13.1%) of the allied healthcare professionals completed the questionnaire. The results revealed that most of the physicans and allied healthcare professionals were in the preparation stage, while most of the nurses were in the precontemplation stage. Multinominal logistic regression analyses showed that, using the precontemplation stage as a reference value, the odds of being in the contemplation stage were lower among participants who were nurses or male or who had a master's or doctoral degree (p < .05). Further, the odds of being in the preparation stage were lower among nurses (p < .01). The odds of being in the maintenance stage were higher among participants that: lived in the Taitung area, were male, or were single (p < .01). The findings provide valuable references for future hospital leaders to develop effective individual physical activity programs for healthcare professionals.


TRPV channel expression in human skin and possible role in thermally induced cell death.
Cell death via necrosis and apoptosis is a hallmark of deep dermal to full-thickness cutaneous burn injuries. Keratinocytes might act as thermosensory cells that transmit information regarding ambient temperature via heat-gated transient receptor potential vanilloid (TRPV) ion channels. The aim of this study was to investigate the distribution of TRPV1, 2, 3, and 4 in uninjured and thermally burned skin. The authors investigated warmth-evoked currents in keratinocytes and cell kinetics of thermally injured keratinocytes in culture with agonists and antagonists of TRPV channels. Specimens of uninjured normal skin and discarded tissue of thermally injured skin were stained for TRPV1, 2, 3, and 4. Cultured primary human keratinocytes were heated for 5 minutes at the following temperatures: 37°C (control), 42°C, and 60°C and thereafter cultured for 24 or 48 hours at 37°C. Thermally stressed cells were treated with TRPV antagonists capsazepine or ruthenium red, and cell viability capacity was determined. TRPV1, TRPV2, TRPV3, and TRPV4 immunoreactivity was differentially identified on basal and suprabasal keratinocytes of healthy human skin. Patch clamp analysis showed a functional response of human keratinocytes at temperatures >40°C. Cell death of keratinocytes after heating at 42°C was reduced by 15 and 5% with ruthenium red and by 20 and 30% by capsazepine at 24 and 48 hours, respectively. Cell death after treatment at 60°C was significantly reduced at 24 hours with capsazepine (22%) or ruthenium red (18%) but only minimally affected after 48 hours postinjury. Interaction with TRPV channels on keratinocytes may offer a new strategy to counteract cell death after thermal injury.


Fog Computing Enabling Industrial Internet of Things: State-of-the-Art and Research Challenges.
Industry is going through a transformation phase, enabling automation and data exchange in manufacturing technologies and processes, and this transformation is called Industry 4.0. Industrial Internet-of-Things (IIoT) applications require real-time processing, near-by storage, ultra-low latency, reliability and high data rate, all of which can be satisfied by fog computing architecture. With smart devices expected to grow exponentially, the need for an optimized fog computing architecture and protocols is crucial. Therein, efficient, intelligent and decentralized solutions are required to ensure real-time connectivity, reliability and green communication. In this paper, we provide a comprehensive review of methods and techniques in fog computing. Our focus is on fog infrastructure and protocols in the context of IIoT applications. This article has two main research areas: In the first half, we discuss the history of industrial revolution, application areas of IIoT followed by key enabling technologies that act as building blocks for industrial transformation. In the second half, we focus on fog computing, providing solutions to critical challenges and as an enabler for IIoT application domains. Finally, open research challenges are discussed to enlighten fog computing aspects in different fields and technologies.


External control of reactions in microdroplets.
Scale reduction of chemical reactions enables novel screening and synthesis approaches that facilitate a highly parallelized and combinatorial exploration of chemical space. Droplet-based microfluidics have evolved as a powerful platform to allow many chemical reactions within small volumes that each can be controlled and manipulated. A significant technical challenge is the ability to change the concentration of reactants inside a droplet. Here we describe a strategy that relies on the use of reactants that are soluble in both oil and water and allow a passive, diffusive exchange of reactants between the oil and aqueous phases to externally control composition of the droplets. We demonstrate the applicability of our approach by externally changing the pH inside microdroplets without the need for physical manipulation or droplet merging.


Morbidity, survival, and site of recurrence after mediastinal lymph-node dissection versus systematic sampling after complete resection for non-small cell lung cancer.
Mediastinal lymph-node dissection was compared to systematic mediastinal lymph-node sampling in patients undergoing complete resection for non-small cell lung cancer with respect to morbidity, duration of chest tube drainage and hospitalization, survival, disease-free survival, and site of recurrence. A consecutive series of one hundred patients with non-small-cell lung cancer, clinical stage T1-3 N0-1 after standardized staging, was divided into two groups of 50 patients each, according to the technique of intraoperative mediastinal lymph-node assessment (dissection versus sampling). Mediastinal lymph-node dissection consisted of removal of all lymphatic tissues within defined anatomic landmarks of stations 2-4 and 7-9 on the right side, and stations 4-9 on the left side according to the classification of the American Thoracic Society. Systematic mediastinal lymph-node sampling consisted of harvesting of one or more representative lymph nodes from stations 2-4 and 7-9 on the right side, and stations 4-9 on the left side. All patients had complete resection. A mean follow-up time of 89 months was achieved in 92 patients. The two groups of patients were comparable with respect to age, gender, performance status, tumor stage, histology, extent of lung resection, and follow-up time. No significant difference was found between both groups regarding the duration of chest tube drainage, hospitalization, and morbidity. However, dissection required a longer operation time than sampling (179 +/- 38 min versus 149 +/- 37 min, p < 0.001). There was no significant difference in overall survival between the two groups; however, patients with stage I disease had a significantly longer disease-free survival after dissection than after sampling (60.2 +/- 7 versus 44.8 +/- 8 months, p < 0.03). Local recurrence was significantly higher after sampling than after dissection in patients with stage I tumor (12.5% versus 45%, p = 0.02) and in patients with nodal tumor negative mediastinum (N0/N1 disease) (46% versus 13%, p = 0.004). Our results suggest that mediastinal lymph-node dissection may provide a longer disease-free survival in stage I non-small cell lung cancer and, most importantly, a better local tumor control than mediastinal lymph-node sampling after complete resection for N0/N1 disease without leading to increased morbidity.


A prospective study of the presentation and management of dancing eye syndrome/opsoclonus-myoclonus syndrome in the United Kingdom.
The incidence, mode of presentation and management of Dancing Eye Syndrome/Opsoclonus-Myoclonus Syndrome (DES/OMS) was prospectively evaluated in 20 United Kingdom (UK) paediatric neurology centres by questionnaire over a 24-month period between 2003 and 2005. Nineteen children were notified, giving an incidence of 0.18 cases per million total population per year. Mean age at presentation was 18 months (range 3-42 months). Fifteen families consented to participate in the study. Atypical features were present in 6/15 cases including very delayed presentation of opsoclonus, dysphagia, and rapid spontaneous improvement without treatment. Only 4/15 cases were associated with neuroblastoma (NB) but current practice in excluding this is diverse and a standardised approach is suggested.


Comparison between standard heparin and tinzaparin for haemodialysis catheter lock.
Tinzaparin offers some advantages over unfractionated heparin (UFH) for hemodialysis circuit anticoagulation. No study has compared these two molecules as hemodialysis catheter locks. This study aimed to compare the efficacy of tinzaparin and UFH as locks for hemodialysis tunnelled central venous catheter in patients with end-stage renal disease (ESRD). A randomized, prospective, single-blinded, controlled study was undertaken. Patients were randomly assigned to receive UFH and tinzaparin for two 7-week periods in a crossover fashion. The doses used were 5,000 U of UFH and 2,000 U of tinzaparin per catheter line. The primary outcome was the need for thrombolytic catheter lock use defined with the Hemodialysis Unit alteplase protocol. Forty-two patients with ESRD were enrolled, totalling 815 UFH lock sessions and 729 tinzaparin lock sessions. A 47.4% reduction in the incidence of alteplase lock use was observed with tinzaparin lock (3.16% vs. 6.01%, chi-square, p = 0.0078). There was no significant difference in the time to first alteplase use between the two locks by Kaplan-Meier survival analysis (logrank, p = 0.0900). Our results suggest that tinzaparin could be an appropriate alternative for a hemodialysis tunnelled central venous catheter lock, but these results should be confirmed with a larger trial.


Wavefront measurement for a hard-X-ray nanobeam using single-grating interferometry.
Wavefront measurement for a hard-X-ray nanobeam using single-grating interferometry based on the Talbot effect and the Fourier transform method was demonstrated in the 1-km-long beamline of SPring-8. 10 keV X-rays were one-dimensionally focused down to 32 nm using a total-reflection elliptical mirror. An intentionally distorted wavefront was generated using a deformable mirror placed just upstream of the focusing mirror. The wavefront measured by interferometry was cross-checked with the phase retrieval method using intensity profiles around the beam waist. Comparison of the obtained wavefront errors revealed that they are in good agreement with each other and with the wavefront error estimated from the shape of the deformable mirror at a ~0.5 rad level.


Adult nephron-specific MR-deficient mice develop a severe renal PHA-1 phenotype.
Aldosterone is the main mineralocorticoid hormone controlling sodium balance, fluid homeostasis, and blood pressure by regulating sodium reabsorption in the aldosterone-sensitive distal nephron (ASDN). Germline loss-of-function mutations of the mineralocorticoid receptor (MR) in humans and in mice lead to the "renal" form of type 1 pseudohypoaldosteronism (PHA-1), a case of aldosterone resistance characterized by salt wasting, dehydration, failure to thrive, hyperkalemia, and metabolic acidosis. To investigate the importance of MR in adult epithelial cells, we generated nephron-specific MR knockout mice (MR(Pax8/LC1)) using a doxycycline-inducible system. Under standard diet, MR(Pax8/LC1) mice exhibit inability to gain weight and significant weight loss compared to control mice. Interestingly, despite failure to thrive, MR(Pax8/LC1) mice survive but develop a severe PHA-1 phenotype with higher urinary Na(+) levels, decreased plasma Na(+), hyperkalemia, and higher levels of plasma aldosterone. This phenotype further worsens and becomes lethal under a sodium-deficient diet. Na(+)/Cl(-) co-transporter (NCC) protein expression and its phosphorylated form are downregulated in the MR(Pax8/LC1) knockouts, as well as the αENaC protein expression level, whereas the expression of glucocorticoid receptor (GR) is increased. A diet rich in Na(+) and low in K(+) does not restore plasma aldosterone to control levels but is sufficient to restore body weight, plasma, and urinary electrolytes. In conclusion, MR deletion along the nephron fully recapitulates the features of severe human PHA-1. ENaC protein expression is dependent on MR activity. Suppression of NCC under hyperkalemia predominates in a hypovolemic state.


The Enriching Interplay between Openness and Interest: A Theoretical Elaboration of the OFCI Model and a First Empirical Test.
The Openness-Fluid-Crystallized-Intelligence (OFCI) model posits long-term relations between Openness and cognitive abilities and has been successfully tested with longitudinal data. However, research on the developmental interplay between cognitive abilities and personality exists only sparsely. The current paper focuses on a theoretical development of the OFCI model which suggests micro-level mechanisms underlying the long-term development. Specifically, within-situation relations between Openness, interests, situational perception, cognitive abilities, and emotions are proposed to explain longitudinal relations between Openness and cognitive abilities. Using experience sampling, selected parts of this elaboration were empirically scrutinized in a first test of the proposed ideas. Openness and specific interest both varied substantially across situations and covaried systematically. In interaction with an indicator of fluid intelligence, this covariation was related to an indicator of crystallized intelligence. The paper contributes to theorizing the intertwined development of personality and cognitive abilities, and highlights the importance of within-situation research for explaining long-term development.


Reference-free detection of isolated SNPs.
Detecting single nucleotide polymorphisms (SNPs) between genomes is becoming a routine task with next-generation sequencing. Generally, SNP detection methods use a reference genome. As non-model organisms are increasingly investigated, the need for reference-free methods has been amplified. Most of the existing reference-free methods have fundamental limitations: they can only call SNPs between exactly two datasets, and/or they require a prohibitive amount of computational resources. The method we propose, discoSnp, detects both heterozygous and homozygous isolated SNPs from any number of read datasets, without a reference genome, and with very low memory and time footprints (billions of reads can be analyzed with a standard desktop computer). To facilitate downstream genotyping analyses, discoSnp ranks predictions and outputs quality and coverage per allele. Compared to finding isolated SNPs using a state-of-the-art assembly and mapping approach, discoSnp requires significantly less computational resources, shows similar precision/recall values, and highly ranked predictions are less likely to be false positives. An experimental validation was conducted on an arthropod species (the tick Ixodes ricinus) on which de novo sequencing was performed. Among the predicted SNPs that were tested, 96% were successfully genotyped and truly exhibited polymorphism.


PREDICTING INDIVIDUAL WELL-BEING THROUGH THE LANGUAGE OF SOCIAL MEDIA.
We present the task of predicting individual well-being, as measured by a life satisfaction scale, through the language people use on social media. Well-being, which encompasses much more than emotion and mood, is linked with good mental and physical health. The ability to quickly and accurately assess it can supplement multi-million dollar national surveys as well as promote whole body health. Through crowd-sourced ratings of tweets and Facebook status updates, we create message-level predictive models for multiple components of well-being. However, well-being is ultimately attributed to people, so we perform an additional evaluation at the user-level, finding that a multi-level cascaded model, using both message-level predictions and userlevel features, performs best and outperforms popular lexicon-based happiness models. Finally, we suggest that analyses of language go beyond prediction by identifying the language that characterizes well-being.


Completeness of the Road-to-Health Booklet and Road-to-Health Card: Results of cross-sectional surveillance at a provincial tertiary hospital.
Accurate record-keeping is important for continuity and quality of care. Completing a child's Road-to-Health Booklet (RTHB), or the older, less detailed, Road-to-Health Card/Chart (RTHC), immediate interpretation thereof and appropriate action facilitates comprehensive care, which could contribute to a decline in child morbidity and mortality. This study aimed to assess the extent to which healthcare personnel working in catchment clinics of Kalafong Provincial Tertiary Hospital (KPTH), Tshwane district, South Africa, complete HIV-related, sociodemographic, neonatal, growth and immunisation information in the RTHC and/or RTHB. A cross-sectional, quantitative record review was conducted. Data were extracted from 318 RTHCs and/or RTHBs of children attending KPTH for paediatric care. Data extraction focused on six main areas, namely documentation of HIV-related, neonatal, sociodemographic, anthropometric, immunisation and vitamin A-related information. During data analysis, age-appropriate completeness scores were generated for each area and completeness of documentation in the RTHB and RTHC was assessed. Data demonstrate significantly less unrecorded HIV-related information (maternal HIV status, timing of maternal HIV testing, timing of maternal antiretroviral therapy [ART] initiation, current maternal ART use and infant feeding decisions) in RTHBs compared with RTHCs (p < 001). Despite this, 24% of all RTHBs had no record of maternal HIV status and 67% of RTHBs from documented HIV-exposed infants had no record of maternal ART duration. Neonatal information completeness was similar between RTHBs and RTHCs, but socio-demographic completeness was significantly better in RTHBs compared with RTHCs (p = 0.006). Growth (especially weight), immunisation and vitamin A completeness was > 80% and similar between RTHBs and RTHCs. Length-for-age, weight-for-length and head circumference were plotted in < 5% of RTHBs and none of the RTHCs. Although completeness of key HIV-related information was better in RTHBs compared with RTHCs, RTHB completeness was suboptimal. Healthcare personnel need reminders to utilise the RTHB optimally to improve continuity and quality of child healthcare.


Switching From Conventional Photodynamic Therapy to Daylight Photodynamic Therapy For Actinic Keratoses: Systematic Review and Meta-analysis.
Actinic keratosis is a precursor lesion to the most common nonmelanoma skin cancer. Conventional photodynamic therapy (PDT) has been shown to be effective, but the procedure is time-consuming, can be very painful, and requires infrastructure. These shortcomings led to the emergence of daylight PDT. To obtain a global estimate of efficacy, we undertook a systematic literature review and performed a meta-analysis of the available evidence on the efficacy and safety of daylight PDT as compared to conventional PDT in the treatment of actinic keratosis and/or field cancerization. The conclusion is that the difference in efficacy is clinically negligible (global estimate of the mean response rate difference, -3.69%; 95% CI, -6.54% to -0.84%). The adverse effects of daylight PDT are mild and localized (79% of patients report no discomfort), and patients report less pain (P<.001). Daylight PDT gives good to excellent cosmetic results in more than 90% of patients, and patient satisfaction is greater (P<.001).


Patient-provider communication and self-care behavior among type 2 diabetes patients.
The purpose of this study was to determine whether diabetes self-care behaviors mediate the association between patient-provider communication (PPC) and diabetes outcomes. An ethnically diverse sample of 752 patients with type 2 diabetes was recruited from 3 different health systems. Subjects were interviewed by telephone about their general and diabetes-specific PPC, diabetes self-care behaviors (medication use, glucose testing, eating, and physical activity), and functional status (physical and mental). Glycemic control was assessed with the HbA1c assay. Statistical tests of the hypothetical path model were adjusted for age, gender, marital status, income, ethnicity, medication regimen, and number of comorbid conditions. General PPC was associated with mental functioning and physical functioning, and diabetes-specific PPC was associated with glycemic control. However, these associations were not mediated by self-care behaviors, which were independently associated with diabetes outcomes. Although self-care and PPC are both important determinants of diabetes patients' health status, these effects are independent. Thus, attempts to improve diabetes outcomes must focus on both good communication and the direct enhancement of illness self-management behaviors.


Hematopoietic niche drives FLT3-ITD acute myeloid leukemia resistance to quizartinib via STAT5-and hypoxia-dependent upregulation of AXL.
Internal tandem duplication in Fms-like tyrosine kinase 3 (FLT3-ITD) is the most frequent mutation observed in acute myeloid leukemia (AML) and correlates with poor prognosis. FLT3 tyrosine kinase inhibitors are promising for targeted therapy. Here, we investigated mechanisms dampening the response to the FLT3 inhibitor quizartinib, which is specific to the hematopoietic niche. Using AML primary samples and cell lines, we demonstrate that convergent signals from the hematopoietic microenvironment drive FLT3-ITD cell resistance to quizartinib through the expression and activation of the tyrosine kinase receptor AXL. Indeed, cytokines sustained phosphorylation of the transcription factor STAT5 in quizartinib-treated cells, which enhanced AXL expression by direct binding of a conserved motif in its genomic sequence. Likewise, hypoxia, another well-known hematopoietic niche hallmark, also enhanced AXL expression. Finally, in a xenograft mouse model, inhibition of AXL significantly increased the response of FLT3-ITD cells to quizartinib exclusively within a bone marrow environment. These data highlight a new bypass mechanism specific to the hematopoietic niche that hampers the response to quizartinib through combined upregulation of AXL activity. Targeting this signaling offers the prospect of a new therapy to eradicate resistant FLT3-ITD leukemic cells hidden within their specific microenvironment, thereby preventing relapses from FLT3-ITD clones.


Localization of agonist-sensitive PtdIns(3,4,5)P3 reveals a nuclear pool that is insensitive to PTEN expression.
Phosphatidylinositol (3,4,5) trisphosphate [PtdIns(3,4,5)P3] is a lipid second messenger, produced by Type I phosphoinositide 3-kinases (PI 3-kinases), which mediates intracellular responses to many growth factors. Although PI 3-kinases are implicated in events at both the plasma membrane and intracellular sites, including the nucleus, direct evidence for the occurrence of PtdIns(3,4,5)P3 at non-plasma membrane locations is limited. We made use of the pleckstrin homology (PH) domain of general receptor for phosphoinositides (Grp1) to detect PtdIns(3,4,5)P3 in an on-section labeling approach by quantitative immunogold electron microscopy. Swiss 3T3 cells contained low levels of PtdIns(3,4,5)P3 that increased up to 15-fold upon stimulation with platelet-derived growth factor (PDGF). The signal was sensitive to PI 3-kinase inhibitors and present mainly at plasma membranes, including lamellipodia, and in a surprisingly large pool within the nuclear matrix. Comparatively little labeling was observed in endomembranes. A similar distribution of PtdIns(3,4,5)P3 was observed in U87MG cells, which lack the PtdIns(3,4,5)P3 phosphatase, PTEN. Re-expression of PTEN into U87MG cells ablated plasma membrane PtdIns(3,4,5)P3, but not the nuclear pool of this lipid even when PTEN was targeted to nuclei. These data have important implications for the versatility of PI 3-kinase signaling and for the proposed functions of PTEN in the nucleus.


Cross-linked nano-onions of carbon nitride in the solid phase: existence of a novel C(48)N(12) aza-fullerene.
We report a new fullerenelike material consisting of cross-linked nano-onions of C and N. Growth of the onion shells takes place atom by atom on a substrate surface and yields thin solid films during magnetron sputter deposition. Electron microscopy and energy loss spectroscopy show that the core shell contains up to 20 at. % N corresponding to C(48)N(12) aza-fullerene composition. Nanoindentation of this nanostructured material gives high resilience with hardness 7 GPa, Young's modulus 37 GPa, and complete elastic recovery after loading with 0.5 mN to a depth of 75 nm. Total energy calculations show the stability of C(60-2n)N(2n) aza-fullerenes and suggest the existence of a novel C(48)N(12) molecule.


Distance estimation in the third dimension in desert ants.
Desert ants of the genus Cataglyphis perform large-scale foraging excursions from which they return to their nest by path integration. They do so by integrating courses steered and the distances travelled into a continually updated home vector. While it is known that the angular orientation is based on skylight cues, it still is largely enigmatic how the ants measure distances travelled. We extended the ants' task into the third dimension by training them to walk within an array of uphill and downhill channels, and later testing them on flat terrain, or vice versa. In these tests the ants indicated homing distances that did not correspond to the distances actually travelled, but to the ground distances; that is, to the sum of the horizontal projections of the uphill and downhill segments of the ants' paths. These results suggest a much more sophisticated mechanism of distance estimation than hitherto thought. The ants must be able to measure the slopes of undulating terrain and to integrate this information into their "odometer" for the distance estimation process.


Pore-lining residues identified by single channel SCAM studies in Cx46 hemichannels.
The substituted cysteine accessibility method was applied to single Cx46 hemichannels to identify residues that participate in lining the aqueous pore of channels formed of connexins. Criteria for assignment to the pore included reactivity to sulfydryl-specific methanethiosulfonate (MTS) reagents from both sides of an open hemichannel and observable effects on open channel properties. We demonstrate reactivity to MTS reagents over a stretch of seventeen amino acids, D51 through L35, that constitute segments of E1 and TM1. Qualitatively, the nature of the effects caused by the Cys substitutions alone and their modification with MTS reagents of either charge indicate side chain valence is most influential in determining single channel properties with D51 and L35 defining the extracellular and intracellular limits, respectively, of the identified pore-lining region. A number of Cys substitutions beyond L35 in TM1 caused severe alterations in hemichannel function and precluded assignment to the pore. Although all six subunits can be modified by smaller MTS reagents, modifications appear limited to fewer subunits with larger reagents.


Interethnic differences in the humoral response to non-repetitive regions of the Plasmodium falciparum circumsporozoite protein.
We analyzed the humoral immune response to the amino- (amino acids 22-125) and carboxy-terminal (amino acids 289-390) non-repetitive domains of the Plasmodium falciparum circumsporozoite protein (PfCSP) in individuals belonging to three west African ethnic groups (the Fulani, Mossi, and Rimaibé) living in the same conditions of hyperendemic transmission in a Sudan savanna area of Burkina Faso. Previous surveys conducted in the same area showed obvious interethnic differences in the susceptibility and immune reactivity to malaria, with the Fulani showing lower infection and disease rates and higher humoral responses to various P. falciparum antigens than sympatric ethnic groups. A total of 764 subjects (311 Mossi, 273 Rimaibé, and 180 Fulani) of all age classes were tested. The total mean +/- SE anti-(CSPf-N-term) and anti-(CSPf-C-term) seroprevalences were 65.6 +/- 1.7% and 57.0 +/- 1.8%, respectively. These seroprevalences were lower than that recorded in the same sample for the central (NANP)40 repetitive domain (88.3 +/- 1.2%). As previously reported for other P. falciparum antigens (PfCSP-(NANP)40, thrombospondin-related anonymous protein, merozoite surface protein-1, Pf155-ring-infected erythrocyte surface antigen, and Pf332), in spite of similar exposure to malaria, the Fulani showed higher immune reactivity than sympatric populations for both antigens tested. Our results confirm the presence of B cell epitopes in the non-repetitive regions of the PfCSP; moreover a further evidence of interethnic differences in the capacity to mount humoral responses against P. falciparum malaria was obtained. The assessment of the biological basis of interethnic heterogeneities in the susceptibility and in the humoral immune responses to malaria appears relevant in the development of anti-malaria vaccines.


Chagas disease in the 21st century: a public health success or an emerging threat?
Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is a major public health burden in Latin America and a potentially serious emerging threat to a number of countries throughout the world. Although public health programs have significantly reduced the prevalence of Chagas disease in Latin America in recent decades, the number of infections in the United States and non-endemic countries in Europe and the Western Pacific Region continues to rise. Moreover, there is still no vaccine or highly effective cure available for the approximately 10 million people currently infected with T. cruzi, a third of which will develop potentially fatal cardiomyopathy and/or severe digestive tract disorders. As Chagas disease becomes an increasingly globalized public health issue in the twenty-first century, continued attentiveness from governmental and health organizations as well as improved diagnostic tools, expanded surveillance and increased research funding will be required to maintain existing public health successes and stymie the spread of the disease to new areas and populations.


Conversion of one-step to two-step self-etch adhesives for improved efficacy and extended application.
One-step self-etch adhesives have restricted use due to their acid-base incompatibility with autocured composites and their behavior as permeable membranes after polymerization. This study examined the feasibility of their conversion to two-step self-etch adhesives via the adjunctive use of a non-solvented, relatively hydrophobic resin coating. iBond, Xeno III and Adper Prompt were used either in multiple coats, or in a single coat followed by the use of a layer of Scotchbond Multi-Purpose Plus bond resin for coupling to light- and auto-cured composites. Four types of experiments were performed. Bonded specimens were examined with TEM after immersion in an ammoniacal silver nitrate tracer. Fluid flow measurements of iBond were conducted using the two application protocols to compare the permeability of the bonded dentin with the original smear layer. Permeability of vital dentin bonded with both application protocols were compared for the transudation of dentin fluid across the bonded dentin. Microtensile bond strengths of dentin bonded with the two protocols were examined for their compatibility with an auto-cured composite. The results of the four experiments were complementary. iBond and Xeno III exhibited "apparent incompatibility" to auto-cured composites that resulted from their inherent permeability. This was confirmed by the presence of dentin fluid transudate on the adhesive surfaces when they were bonded to vital dentin. Conversely, Adper Prompt exhibited "true incompatibility" to auto-cured composites that was caused by adverse acid-base interaction, masking the inherent permeability of this adhesive. "True" and "apparent" incompatibility issues were eliminated upon their conversion to two-step self-etch adhesives.