Quantitative trait loci for chemical body composition traits in pigs and their positional associations with body tissues, growth and feed intake.
In this study, quantitative trait loci (QTL) for chemical and physical body composition, growth and feed intake in pigs were identified in a three-generation full-sib population, developed by crossing Pietrain sires with a commercial dam line. Phenotypic data from 315 F(2) animals were available for protein and lipid deposition measured in live animals by the deuterium dilution technique at 30-, 60-, 90-, 120- and 140-kg body weight. At 140-kg body weight, carcass characteristics were measured by the AutoFOM grading system and after dissection. Three hundred and eighty-six animals from 49 families were genotyped for 51 molecular markers covering chromosomes SSC2, SSC4, SSC8, SSC9, SSC10 and SSC14. Novel QTL for protein (lipid) content at 60-kg body weight and protein (lipid) accretion from 120 to 140 kg were detected on SSC9 near several previously detected QTL for lean and fat tissue in neck, shoulder and ham cuts. Another QTL for lipid accretion was found on SSC8, closely associated with a QTL for intramuscular fat content. QTL for daily feed intake were detected on SSC2 and SSC10. The favourable allele of a QTL for food conversion ratio (FCR) on SSC2 was associated with alleles for increased lean tissue and decreased fat tissue. Because no QTL for growth rate were found in the region, the QTL for FCR is most likely due to a change in body composition. These QTL provide insights into the genomic regulation of chemical or physical body composition and its association with feed intake, feed efficiency and growth.


Near-roadway pollution and childhood asthma: implications for developing "win-win" compact urban development and clean vehicle strategies.
The emerging consensus that exposure to near-roadway traffic-related pollution causes asthma has implications for compact urban development policies designed to reduce driving and greenhouse gases. We estimated the current burden of childhood asthma-related disease attributable to near-roadway and regional air pollution in Los Angeles County (LAC) and the potential health impact of regional pollution reduction associated with changes in population along major traffic corridors. The burden of asthma attributable to the dual effects of near-roadway and regional air pollution was estimated, using nitrogen dioxide and ozone as markers of urban combustion-related and secondary oxidant pollution, respectively. We also estimated the impact of alternative scenarios that assumed a 20% reduction in regional pollution in combination with a 3.6% reduction or 3.6% increase in the proportion of the total population living near major roads, a proxy for near-roadway exposure. We estimated that 27,100 cases of childhood asthma (8% of total) in LAC were at least partly attributable to pollution associated with residential location within 75 m of a major road. As a result, a substantial proportion of asthma-related morbidity is a consequence of near-roadway pollution, even if symptoms are triggered by other factors. Benefits resulting from a 20% regional pollution reduction varied markedly depending on the associated change in near-roadway proximity. Our findings suggest that there are large and previously unappreciated public health consequences of air pollution in LAC and probably in other metropolitan areas with dense traffic corridors. To maximize health benefits, compact urban development strategies should be coupled with policies to reduce near-roadway pollution exposure.


Microstructural integrity of white matter moderates an association between childhood adversity and adult trait anger.
Amongst a number of negative life sequelae associated with childhood adversity is the later expression of a higher dispositional tendency to experience anger and frustration to a wide range of situations (i.e., trait anger). We recently reported that an association between childhood adversity and trait anger is moderated by individual differences in both threat-related amygdala activity and executive control-related dorsolateral prefrontal cortex (dlPFC) activity, wherein individuals with relatively low amygdala and high dlPFC activity do not express higher trait anger even when having experienced childhood adversity. Here, we examine possible structural correlates of this functional dynamic using diffusion magnetic resonance imaging data from 647 young adult men and women volunteers. Specifically, we tested whether the degree of white matter microstructural integrity as indexed by fractional anisotropy modulated the association between childhood adversity and trait anger. Our analyses revealed that higher microstructural integrity of multiple pathways was associated with an attenuated link between childhood adversity and adult trait anger. Amongst these pathways was the uncinate fasciculus (UF; ΔR 2 = 0.01), which not only provides a major anatomical link between the amygdala and prefrontal cortex but also is associated with individual differences in regulating negative emotion through top-down cognitive reappraisal. These findings suggest that higher microstructural integrity of distributed white matter pathways including but not limited to the UF may represent an anatomical foundation serving to buffer against the expression of childhood adversity as later trait anger, which is itself associated with multiple negative health outcomes.


Resveratrol inhibits vascular smooth muscle cell proliferation and induces apoptosis.
In France, despite a high intake of dietary cholesterol and saturated fat, the cardiovascular death rate is one of the lowest among developed countries. This "French paradox" has been postulated to be related to the high red wine intake in France. The aim of this study was to determine the effects of resveratrol, a major polyphenol component of red wine, on vascular smooth muscle cell (SMC) proliferation in vitro. SMCs were exposed to 10(-6) to 10(-4) M resveratrol and cell proliferation was assessed by cell counting. Cell cycle analysis was done by treating cells with propidium iodide followed by flow-activated cell sorting. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling staining. We demonstrate that resveratrol inhibited bovine aortic SMC proliferation in a dose-dependent manner. The lowest concentration of resveratrol resulting in a significant decrease in SMC proliferation compared with control was 10(-5) M. By flow cytometry, we observed a block in the G1-S phase of the SMC cycle. Resveratrol treatment also resulted in a dose-dependent apoptosis of SMCs but had no effects on SMC morphology. The results indicated that vascular SMC proliferation could be inhibited by resveratrol through a block on G1-S phase and by an increase in apoptosis. It supports the conjecture that red wine consumption may have a beneficial effect on cardiovascular mortality. Our results suggest that resveratrol inhibits, in a dose-dependent manner, smooth muscle cell proliferation, which may help to partially explain a beneficial effect of wine drinking. This inhibition is related to an early block in the cell cycle and also to a dose-dependent apoptotic effect. The present study demonstrates that resveratrol not only is an indirect marker of a healthy life style and alimentation but may also be directly responsible for the French paradox.


p16INK4a and its regulator miR-24 link senescence and chondrocyte terminal differentiation-associated matrix remodeling in osteoarthritis.
Recent evidence suggests that tissue accumulation of senescent p16INK4a-positive cells during the life span would be deleterious for tissue functions and could be the consequence of inherent age-associated disorders. Osteoarthritis (OA) is characterized by the accumulation of chondrocytes expressing p16INK4a and markers of the senescence-associated secretory phenotype (SASP), including the matrix remodeling metalloproteases MMP1/MMP13 and pro-inflammatory cytokines interleukin-8 (IL-8) and IL-6. Here, we evaluated the role of p16INK4a in the OA-induced SASP and its regulation by microRNAs (miRs). We used IL-1-beta-treated primary OA chondrocytes cultured in three-dimensional setting or mesenchymal stem cells differentiated into chondrocyte to follow p16INK4a expression. By transient transfection experiments and the use of knockout mice, we validate p16INK4a function in chondrocytes and its regulation by one miR identified by means of a genome-wide miR-array analysis. p16INK4a is induced upon IL-1-beta treatment and also during in vitro chondrogenesis. In the mouse model, Ink4a locus favors in vivo the proportion of terminally differentiated chondrocytes. When overexpressed in chondrocytes, p16INK4a is sufficient to induce the production of the two matrix remodeling enzymes, MMP1 and MMP13, thus linking senescence with OA pathogenesis and bone development. We identified miR-24 as a negative regulator of p16INK4a. Accordingly, p16INK4a expression increased while miR-24 level was repressed upon IL-1-beta addition, in OA cartilage and during in vitro terminal chondrogenesis. We disclosed herein a new role of the senescence marker p16INK4a and its regulation by miR-24 during OA and terminal chondrogenesis.


Optimal amounts of fluorescent dye improve expression microarray results in tumor specimens.
Expression microarrays have great potential for clinical use but variability of the results represents a challenge for reliable practical application. The amount of fluorescent dye used in microarray experiments is a significant source of variability that has not been systematically studied. Here we demonstrate that the quantity of Cy3 dye affects microarray results performed on tumor specimens. Signal-to-noise ratios and coefficients of variation are significantly improved by increasing Cy3 to 150-180 pmol, but any further increase does not improve the data. In conclusion, optimal amounts of dye reduce variability and improve reliability of expression microarray experiments.


Agonistic monoclonal antibody against CD40 receptor decreases lymphocyte apoptosis and improves survival in sepsis.
Sepsis causes a marked apoptosis-induced depletion of lymphocytes. The degree of lymphocyte apoptosis during sepsis strongly correlates with survival. CD40, a member of the TNFR family, is expressed on APCs and has potent antiapoptotic activity. In this study we determined whether an agonistic Ab against CD40 could protect lymphocytes from sepsis-induced apoptosis. Secondly, we examined potential antiapoptotic mechanisms of the putative protection. Lastly, we aimed to determine whether anti-CD40 treatment could improve survival in sepsis. CD1 mice were made septic by the cecal ligation and puncture method and treated postoperatively with anti-CD40 Ab. Treatment with anti-CD40 completely abrogated sepsis-induced splenic B cell death and, surprisingly, decreased splenic and thymic T cell death as well (p < 0.001). To investigate the mechanism of protection of anti-CD40 therapy on T cells, CD40 receptor expression was examined. As anticipated, the CD40 receptor was constitutively expressed on B cells, but, unexpectedly, splenic and thymic T cells were found to express CD40 receptor during sepsis. Furthermore, CD4+CD8- T cells were the predominant subtype of T cells expressing CD40 receptor during sepsis. Additionally, the antiapoptotic protein Bcl-x(L) was found to be markedly increased in splenic B and T cells as well as in thymic T cells after treatment with anti-CD40 Ab (p < 0.0025). Lastly, mice that were made septic in a double injury model of sepsis had improved survival after treatment with anti-CD40 as compared with controls (p = 0.05). In conclusion, anti-CD40 treatment increases Bcl-x(L), provides nearly complete protection against sepsis-induced lymphocyte apoptosis, and improves survival in sepsis.


Pharmacists' knowledge, attitudes, and practices regarding influenza vaccination and treatment of pregnant women.
To understand pharmacists' knowledge, attitudes, and practices regarding vaccination and treatment of pregnant women for seasonal influenza and pandemic 2009 influenza A (H1N1). Descriptive, exploratory, nonexperimental study. United States between January 21, 2010, and February 9, 2010. 606 pharmacists who participated in the American Pharmacists Association (APhA) Immunization Certificate Training Program and practice in chain, supermarket, mass merchandise, and independent pharmacies. Electronic survey sent by APhA to 7,356 pharmacists who had participated in its Immunization Certificate Training Program. Pharmacists' knowledge, attitudes, and practices regarding vaccination and antiviral treatment of pregnant women for seasonal and H1N1 influenza. Respondents were more likely to recognize that pregnant women are at an increased risk associated with H1N1 influenza (85%) than to recognize the increased risk associated with seasonal influenza (78%). However, respondents were less likely to believe that they have an important role in vaccinating pregnant women compared with the general public (82% vs. 97%) and less likely to agree that 2009 H1N1 vaccine was safe during pregnancy compared with the seasonal influenza vaccine (78% vs. 87%). Pharmacists who had been vaccinated themselves were more likely to recommend vaccination for pregnant patients. Only 38% believed that antiviral medications such as oseltamivir can be given during pregnancy because the benefits outweigh the risks. However, in response to case studies of pregnant women who were candidates for antiviral medications, respondents indicated that they would take extraordinary steps to ensure that pregnant women were either referred for medical assessment or for assistance in obtaining prescribed antiviral medications. Education efforts that focus on the effectiveness and safety of influenza vaccination during pregnancy and the benefits of treating pregnant women with confirmed or suspected influenza with antiviral medications may be useful in improving pharmacists' support of pharmaceutical interventions to reduce the impact of influenza in pregnant women. Pharmacists' personal decisions regarding vaccination may be a marker for their overall assessment of risks and benefits and may influence their recommendations for pregnant patients.


High genetic diversity is not essential for successful introduction.
Some introduced populations thrive and evolve despite the presumed loss of diversity at introduction. We aimed to quantify the amount of genetic diversity retained at introduction in species that have shown evidence of adaptation to their introduced environments. Samples were taken from native and introduced ranges of Arctotheca populifolia and Petrorhagia nanteuilii. Using microsatellite data, we identified the source for each introduction, estimated genetic diversity in native and introduced populations, and calculated the amount of diversity retained in introduced populations. These values were compared to those from a literature review of diversity in native, confamilial populations and to estimates of genetic diversity retained at introduction. Gene diversity in the native range of both species was significantly lower than for confamilials. We found that, on average, introduced populations showing evidence of adaptation to their new environments retained 81% of the genetic diversity from the native range. Introduced populations of P. nanteuilii had higher genetic diversity than found in the native source populations, whereas introduced populations of A. populifolia retained only 14% of its native diversity in one introduction and 1% in another. Our literature review has shown that most introductions demonstrating adaptive ability have lost diversity upon introduction. The two species studied here had exceptionally low native range genetic diversity. Further, the two introductions of A. populifolia represent the largest percentage loss of genetic diversity in a species showing evidence of substantial morphological change in the introduced range. While high genetic diversity may increase the likelihood of invasion success, the species examined here adapted to their new environments with very little neutral genetic diversity. This finding suggests that even introductions founded by small numbers of individuals have the potential to become invasive.


Evidence for a physiological role for nitric oxide in the regulation of the LH surge: effect of central administration of antisense oligonucleotides to nitric oxide synthase.
Antisense oligonucleotides of brain-nitric oxide synthase (b-NOS) and endothelial-NOS (e-NOS) were used in steroid-primed ovariectomized rats to examine the physiological role of nitric oxide in the regulation of the LH surge. Since macrophage-NOS (m-NOS) is not produced in the hypothalamus under normal conditions, the m-NOS antisense oligonucleotide was used as control for the possible toxicity of the phosphorothioated and propynylated antisense oligonucleotides used. Female rats were ovariectomized on day 70 of age and implanted with a third ventricle cannula on day 77 of age, injected with 5 micrograms of estradiol on days 84 and 85 of age at 17.00 h and with 1 mg progesterone or vehicle on day 86 at 09.00 h. Blood samples were collected between 13.00 and 19.00 h on day 86 of age via a jugular cannula inserted on day 85 of age. Antisense oligonucleotides (400 or 800 ng) or vehicle were injected in the third ventricle at 17.00 h on days 84 and 85 just before the estradiol injection and at 06.00 and 12.00 h on day 86. Neither the 400-ng nor the 800-ng dose of m-NOS AS had any effect on the steroid-induced LH surge. In contrast, central administration of the 400-ng dose of e-NOS AS and the 800-ng dose of b-NOS AS significantly attenuated the steroid-induced LH surge. The 40% reduction in LH by e-NOS AS and b-NOS AS was accompanied by a 33 and 28% reduction in their respective protein levels as shown by Western blots. The higher amount of b-NOS AS needed to reduce the LH surge is probably due to the high abundance of b-NOS in the hypothalamus as compared to e-NOS. As a whole, this study provides significant evidence for a physiological role of nitric oxide in mediating the steroid-induced LH surge.


Awareness and Use of mHealth Apps: A Study from England.
Purpose: Mobile health (mHealth) solutions have become an inevitable element of the healthcare landscape. The recommendation and use of mHealth is important, but it is often underutilised. This study was conducted in England. It aimed to determine the use and recommendation of mHealth apps by pharmacists, the public's perceptions of mHealth apps in general, and the awareness and use of health apps by diabetic patients in particular. Methods: The study used a mixed research approach, utilising a sequence of survey-based questionnaires with pharmacists and the general public, followed by semi-structured interviews with diabetic patients. Results: Pharmacists' questionnaires revealed that 56% of the respondents were aware of health apps, 60% of which recommended them to patients. Over 76% of the individuals owned a smartphone. The types of applications that saw the most use from the general public were health and lifestyle apps (24%), social apps (19%), followed by news (18%). Although eight out of nine diabetic patients owned a smartphone, only three used diabetes apps. Diabetic patients also suggested an interest in using diabetes apps to aid in optimising care via the utilisation of visual aids, reminders, recording patient data, social coaching, and remote collaboration with healthcare professionals (HCPs), but time was seen as the biggest obstacle to using a diabetes mHealth application. Conclusion: Despite the growing number of mHealth apps, the level of awareness and usability of such apps by patients and pharmacists was still relatively low. Nevertheless, the majority who used health apps found them to be beneficial, and the public agreed that it helped them to live a healthier lifestyle. Therefore, health apps have great potential in health promotion. Pharmacists are ideally placed to promote them and make patients more aware of them. To increase the use of these apps, it is necessary to first increase awareness and knowledge of these apps, both to the public and to healthcare professionals.


Pemetrexed, Bevacizumab, or the Combination As Maintenance Therapy for Advanced Nonsquamous Non-Small-Cell Lung Cancer: ECOG-ACRIN 5508.
Pemetrexed or bevacizumab is used for maintenance therapy of advanced nonsquamous non-small-cell lung cancer (NSCLC). The combination of bevacizumab and pemetrexed has also demonstrated efficacy. We conducted a randomized study to determine the optimal maintenance therapy. Patients with advanced nonsquamous NSCLC and no prior systemic therapy received carboplatin (area under the curve, 6), paclitaxel (200 mg/m2), and bevacizumab (15 mg/kg) for up to four cycles. Patients without progression after four cycles were randomly assigned to maintenance therapy with bevacizumab (15 mg/kg), pemetrexed (500 mg/m2), or a combination of the two agents. The primary end point was overall survival, with bevacizumab serving as the control group. Of the 1,516 patients enrolled, 874 (57%) were randomly assigned after induction therapy to one of the three maintenance therapy groups. With a median follow-up of 50.6 months, median survival with pemetrexed was 15.9 months, compared with 14.4 months with bevacizumab (hazard ratio [HR], 0.86; P = .12); median survival with pemetrexed and bevacizumab was 16.4 months (HR, 0.9; P = .28); median progression-free survival was 4.2, 5.1 (HR, 0.85; P = .06), and 7.5 months (HR, 0.67; P < .001) for the three groups, respectively. Incidence of worst grade 3 to 4 toxicity was 29%, 37%, and 51%, respectively, for bevacizumab, pemetrexed, and the combination regimen. Single-agent bevacizumab or pemetrexed is efficacious as maintenance therapy for advanced nonsquamous NSCLC. Because of a lack of survival benefit and higher toxicity, the combination of bevacizumab and pemetrexed cannot be recommended.


Noise propagation in linear and nonlinear inverse scattering.
The propagation of noise from the data to the reconstructed speed of sound image by inverse scattering within the framework of the Lippmann-Schwinger integral equation of scattering is discussed. The inversion algorithm that was used consisted in minimizing a Tikhonov functional in the unknown speed of sound. The gradient of the objective functional was computed by the method of the adjoint fields. An analytical expression for the inverse scattering covariance matrix of the image noise was derived. It was shown that the covariance matrix in the linear x-ray computed tomography is a special case of the inverse scattering matrix derived in this paper. The matrix was also analyzed in the limit of the linearized Born approximation, and the results were found to be in qualitative agreement with those recently reported in the literature for Born inversion using filtered backpropagation algorithm. Finally, the applicability of the analysis reported here to the obstacle problem and the physical optics approximation was discussed.


Demographic features of idiopathic macular telangiectasia in Korean patients.
To investigate the clinical and demographic features of idiopathic macular telangiectasia (MacTel) in Korean patients since the introduction of spectral domain optical coherence tomography (SD-OCT). We reviewed medical records of patients who were diagnosed with MacTel from 2009 to 2013. All patients underwent fluorescein angiography and SD-OCT and were classified as type 1 or type 2 according to the classification system proposed by Yannuzzi. Over a period of 5 years, 4 (18.2%) patients were diagnosed with type 1 MacTel and 18 (81.8%) patients were diagnosed with type 2 MacTel. All patients with type1 MacTel were male, and their mean age was 51 ± 8.6 years. Among patients with type 2 MacTel, 3 (16.7%) were male, 15 (83.3%) were female, and the mean age was 60 ± 13.6 years. Whereas all type 1 MacTel patients had either metamorphopsia or mild scotoma, of the 18 patients with type 2 MacTel, only 4 (22.2%) had those symptoms, 10 (55.6%) complained of only mild visual impairment, and the other 4 (22.2%) had no symptoms. Intraretinal cystoid spaces were observed in 26 (72.2%) of 36 eyes with type 2 MacTel by SD-OCT. These cystoid spaces had irregular boundaries and did not correspond to angiographic leakages. Type 2 MacTel was most common in the present study. The wider availability of SD-OCT may have contributed to the diagnosis of type 2 MacTel. Type 2 MacTel may be more prevalent than type 1 in Koreans, which corresponds to the results of Western countries.


Capillary blood flow around microglial somata determines dynamics of microglial processes in ischemic conditions.
Microglia are the resident immune cells in the brain. Under normal conditions, resting ramified microglia constantly extend and retract fine processes while performing immunological surveillance. In ischemia, microglia become activated as demonstrated by morphological changes during deramification leading to transformation from ramified to amoeboid form. In vivo two-photon microscopy of enhanced green fluorescent protein (EGFP)-expressing microglia in mouse neocortex was used to examine microglial dynamics during the early periods of focal and global ischemia. A penumbra-like "area-at-risk" surrounded by a square-shaped area of severely hypoperfused tissue was created by laser-induced photothrombosis. The dynamics of microglial processes in the area-at-risk was strongly correlated with capillary blood flow (BF) measured within 10 μm of microglial somata. Changes in BF around distal microglial processes (>30 μm from somata) had no effect on microglial dynamics. A severe reduction of capillary BF near somata by 84% ± 6% resulted in initiation of microglial deramification, suggesting activation. A moderate decrease in BF near somata by 22% ± 5% or increase by 87% ± 10%, reflecting a redistribution of capillary BF, had no effect on microglial morphology. Complete BF loss during cardiac arrest (CA) or transient bilateral common carotid artery occlusion (BCCAO) entirely stalled all microglial processes without structural changes. Reperfusion after BCCAO induced recovery of microglial dynamics to preocclusion values. These findings suggest that during ischemia, the severe drop in BF around microglial somata coincides with morphological activation. However, this activation requires some residual BF, because complete perfusion loss (as during BCCAO and CA) did not support microglial deramification.


Cannabis use and deliberate self-harm in adolescence: a comparative analysis of associations in England and Norway.
The objective of this study was to test hypotheses on causality and selection regarding associations between cannabis use and deliberate self-harm (DSH) among adolescents. School surveys were conducted among 9,800 adolescents in England and Norway applying identical measures on deliberate self-harm, suicidal thoughts, cannabis use, and various potential confounders. Cannabis use was more prevalent in England than in Norway. It was associated with DHS, suicidal thoughts and various risk factors for DSH. However, these associations were stronger in Norway than in England. The adjusted associations between cannabis use and suicidal thoughts were non-significant in both countries. The adjusted cannabis-DSH association was non-significant in England but significant in Norway. Elevated risk of DSH in adolescent cannabis users seems to be mainly due to selection mechanisms. Thus the association is not likely to be direct but due to other shared contributory factors.


Pharmacokinetics, Pharmacodynamics, and PKPD Modeling of Curcumin in Regulating Antioxidant and Epigenetic Gene Expression in Healthy Human Volunteers.
Curcumin is a major component of the spice turmeric ( Curcuma longa), often used in food or as a dietary supplement. Many preclinical studies on curcumin suggest health benefits in many diseases due to its antioxidant/anti-inflammatory and epigenetic effects. The few human studies and curcumin's unfavorable pharmacokinetics (PK) have limited its potential, leading researchers to study and develop formulations to improve its PK. The purpose of this clinical study is to describe the acute pharmacokinetics and pharmacodynamics (PK/PD) of commercially marketed curcumin in normal, healthy human volunteers. Twelve volunteers received a 4 g dose of curcumin capsules with a standard breakfast. Plasma samples were collected at specified time points and analyzed for curcumin and its glucuronide levels. RNA was extracted from leukocytes and analyzed for expression of select antioxidant and epigenetic histone deacetylase (HDAC) genes. Plasma levels of parent curcumin were below the detection limit by HPLC-ITMS/MS/MS. However, curcumin-O-glucuronide (COG), a major metabolite of curcumin, was detected as soon as 30 min. These observations of little to no curcumin and some levels of metabolite are in line with previous studies. PD marker antioxidant genes NRF2, HO-1, and NQO1 and epigenetic genes HDAC1, HDAC2, HDAC3, and HDAC4 were quantified by qPCR. COG PK is well-described by a one-compartment model, and the PK/PD of COG and its effect on antioxidant and epigenetic gene expression are captured by an indirect response model (IDR). A structural population PK model was sequentially established using a nonlinear mixed-effect model program (Monolix Lixoft, Orsay, France). Physiologically based pharmacokinetic modeling (PBPK) and simulation using Simcyp correlated well with the observed data. Taken together, these results show that the bioavailability of the parent curcumin compound is low, and oral administration of curcumin can still deliver detectable levels of curcumin glucuronide metabolite. But most importantly, it elicits antioxidant and epigenetic effects which could contribute to the overall health beneficial effects of curcumin.


Interspecific competition during transmission of two sympatric malaria parasite species to the mosquito vector.
The role of species interactions in structuring parasite communities remains controversial. Here, we show that interspecific competition between two avian malaria parasite species, Plasmodium gallinaceum and P. juxtanucleare, occurs as a result of interference during parasite fertilization within the bloodmeal of the mosquito. The significant reduction in the transmission success of P. gallinaceum to mosquitoes, due to the co-infecting P. juxtanucleare, is predicted to have compromised its colonization of regions occupied by P. juxtanucleare and, thus, may have contributed to the restricted global distribution of P. gallinaceum. Such interspecies interactions may occur between human malaria parasites and, thus, impact upon parasite species epidemiology, especially in regions of seasonal transmission.


Annotated chemical patent corpus: a gold standard for text mining.
Exploring the chemical and biological space covered by patent applications is crucial in early-stage medicinal chemistry activities. Patent analysis can provide understanding of compound prior art, novelty checking, validation of biological assays, and identification of new starting points for chemical exploration. Extracting chemical and biological entities from patents through manual extraction by expert curators can take substantial amount of time and resources. Text mining methods can help to ease this process. To validate the performance of such methods, a manually annotated patent corpus is essential. In this study we have produced a large gold standard chemical patent corpus. We developed annotation guidelines and selected 200 full patents from the World Intellectual Property Organization, United States Patent and Trademark Office, and European Patent Office. The patents were pre-annotated automatically and made available to four independent annotator groups each consisting of two to ten annotators. The annotators marked chemicals in different subclasses, diseases, targets, and modes of action. Spelling mistakes and spurious line break due to optical character recognition errors were also annotated. A subset of 47 patents was annotated by at least three annotator groups, from which harmonized annotations and inter-annotator agreement scores were derived. One group annotated the full set. The patent corpus includes 400,125 annotations for the full set and 36,537 annotations for the harmonized set. All patents and annotated entities are publicly available at www.biosemantics.org.


The minimal active human SVA retrotransposon requires only the 5'-hexamer and Alu-like domains.
RNA-based duplication mediated by reverse transcriptase (RT), a process termed retrotransposition, is ongoing in humans and is a source of significant inter- and perhaps intraindividual genomic variation. The long interspersed element 1 (LINE-1 or L1) ORF2 protein is the genomic source for RT activity required for mobilization of its own RNA in cis and other RNAs, such as SINE/variable-number tandem-repeat (VNTR)/Alu (SVA) elements, in trans. SVA elements are ~2-kb hominid-specific noncoding RNAs that have resulted in single-gene disease in humans through insertional mutagenesis or aberrant mRNA splicing. Here, using an SVA retrotransposition cell culture assay in U2OS cells, we investigated SVA domains important in L1-mediated SVA retrotransposition. Partial- and whole-domain deletions revealed that removal of either the Alu-like or SINE-R domain in the context of a full-length SVA has little to no effect, whereas removal of the CT hexamer or the VNTR domain can result in a 75% decrease in activity. Additional experiments demonstrate that the Alu-like fragment alone can retrotranspose at low levels while the addition of the CT hexamer can enhance activity as much as 2-fold compared to that of the full-length SVA. These results suggest that no SVA domain is essential for retrotransposition in U2OS cells and that the 5' end of SVA (hexamer and Alu-like domain) is sufficient for retrotransposition.


Enabling the extended compact genetic algorithm for real-parameter optimization by using adaptive discretization.
An adaptive discretization method, called split-on-demand (SoD), enables estimation of distribution algorithms (EDAs) for discrete variables to solve continuous optimization problems. SoD randomly splits a continuous interval if the number of search points within the interval exceeds a threshold, which is decreased at every iteration. After the split operation, the nonempty intervals are assigned integer codes, and the search points are discretized accordingly. As an example of using SoD with EDAs, the integration of SoD and the extended compact genetic algorithm (ECGA) is presented and numerically examined. In this integration, we adopt a local search mechanism as an optional component of our back end optimization engine. As a result, the proposed framework can be considered as a memetic algorithm, and SoD can potentially be applied to other memetic algorithms. The numerical experiments consist of two parts: (1) a set of benchmark functions on which ECGA with SoD and ECGA with two well-known discretization methods: the fixed-height histogram (FHH) and the fixed-width histogram (FWH) are compared; (2) a real-world application, the economic dispatch problem, on which ECGA with SoD is compared to other methods. The experimental results indicate that SoD is a better discretization method to work with ECGA. Moreover, ECGA with SoD works quite well on the economic dispatch problem and delivers solutions better than the best known results obtained by other methods in existence.


Role of Angiopoietins and Tie-2 in Diabetic Retinopathy.
The aim of this study was to determine the serum levels of angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), soluble vascular endothelial tyrosine kinase receptor (Tie-2) and vascular endothelial growth factor (VEGF), in the serum of type 2 diabetic patients having non-proliferative (NPDR) or proliferative diabetic retinopathy (PDR). One hundred patients with type 2 diabetes mellitus were involved in this cross-sectional study. They were classified into 25 diabetic patients without retinopathy, 35 patients with NPDR and 40 PDR patients. The study was carried out in the outpatients clinic of the Research Institute of Ophthalmology, Giza, Egypt from August 2016 through May 2017. Serum VEGF, Ang-1, Ang-2 and Tie-2 receptor levels were assayed using enzyme linked immunosorbent assay (ELISA). Data were analyzed by SPSS version 20 and Microsoft Excel (Version 10) using ANOVA, Harman's single factor test, and Pearson's Product Moment Correlation. The serum levels of Ang-2 and VEGF were significantly elevated in NPDR groups and PDR group compared to diabetics without retinopathy groups (p<0.001). The serum levels of Ang-1 were significantly higher in the NPDR group compared to the diabetics without retinopathy groups (p<0.01), while no significant difference was observed between the PDR and diabetics without retinopathy groups. Ang-1/Ang-2 ratio was the lowest in the PDR group compared to the NPDR and diabetics without retinopathy groups. The serum levels of Tie-2 were not significantly changed among the three studied groups, serum Ang-2 was positively correlated with VEGF and Tie-2 in the PDR and NPDR groups. The angiopoietin/Tie system and VEGF are essential features in the commencement and development of PDR.


Reciprocal androgen receptor/interleukin-6 crosstalk drives oesophageal carcinoma progression and contributes to patient prognosis.
Oesophageal squamous cell carcinoma (ESCC), a leading lethal malignancy of the digestive tract, is characterized by marked gender disparity. Clarifying the roles of the function and regulatory pathway of the androgen receptor (AR) will improve our understanding of oesophageal cancer progression, thereby facilitating the personalized management of ESCC. Here we report evidence to show that AR is a key mediator of inflammatory signals in ESCC cancer progression. High AR expression was associated with poor overall survival in tobacco-using ESCC patients but not in ESCC patients not using tobacco. A gain and loss of AR function enhanced and repressed ESCC cell growth, respectively, by altering cell cycle progression. In mice bearing human ESCC xenografts, silencing AR expression attenuated tumour growth, whereas AR overexpression promoted tumour growth in mice of different androgen statuses (male, female, and castrated male). Array assays revealed that the inflammatory cytokine interleukin-6 (IL6) is a prominent AR target gene in ESCC. By directly binding to the IL6 promoter, AR enhances IL6 transcription, and IL6 can in turn activate AR expression, thus forming a reciprocal regulatory circuit to sustain STAT3 oncogenic signalling in ESCC. Moreover, high expression levels of both AR and IL6 in human ESCC predict poor clinical outcome in tobacco users. Together, these data establish that AR promotes ESCC growth and is associated with poor patient prognosis. The discovery of a positive feedback loop between IL6 and AR bridges the knowledge gaps among lifestyle factor-associated inflammation, gender disparity, and oesophageal carcinoma. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Unintended consequences of delisting routine eye exams on retinopathy screening for people with diabetes in Ontario, Canada.
Routine eye examinations for healthy adults aged 20-64 years were delisted from the Ontario Health Insurance Plan in 2004, but they continue to be insured for people with diabetes regardless of age. We sought to assess whether the delisting of routine eye examinations for healthy adults had the unintended consequence of decreasing retinopathy screening for adults with diabetes. We used administrative data to calculate eye examinations for people with diabetes ages 40-64 years and 65 years and older in each 2-year period from 1998 to 2010. We examined differences by sex, income, rurality and type of health care provider. We used segmented linear regression to assess the change in trend before and after 2004. For people with diabetes aged 65 years and older, eye examinations rose gradually from 1998 to 2010, with no substantial change between 2004 and 2006. For people with diabetes aged 40-65 years, there was an 8.7% (95% confidence interval [CI] 6.3%-11.1%) decrease in eye examinations between 2004 and 2006. Results were similar for all population subgroups. Ophthalmologic examinations decreased steadily for both age groups during the study period, and there was a decline in optometry examinations for people ages 40-65 years after 2004. The delisting of routine eye examinations for healthy adults in Ontario had the unintended consequence of reducing publicly funded retinopathy screening for people with diabetes. More research is needed to understand whether patients are being charged for an insured service or to what degree misunderstanding has prevented patients from seeking care.


Changing climate and overgrazing are decimating Mongolian steppes.
Satellite observations identify the Mongolian steppes as a hotspot of global biomass reduction, the extent of which is comparable with tropical rainforest deforestation. To conserve or restore these grasslands, the relative contributions of climate and human activities to degradation need to be understood. Here we use a recently developed 21-year (1988-2008) record of satellite based vegetation optical depth (VOD, a proxy for vegetation water content and aboveground biomass), to show that nearly all steppe grasslands in Mongolia experienced significant decreases in VOD. Approximately 60% of the VOD declines can be directly explained by variations in rainfall and surface temperature. After removing these climate induced influences, a significant decreasing trend still persists in the VOD residuals across regions of Mongolia. Correlations in spatial patterns and temporal trends suggest that a marked increase in goat density with associated grazing pressures and wild fires are the most likely non-climatic factors behind grassland degradation.


Significance of Ovarian Endometriosis on the Prognosis of Ovarian Clear Cell Carcinoma.
The aim of this study was to evaluate the significance of ovarian endometriosis on the prognosis of ovarian clear cell carcinoma (OCCC). Patients with OCCC were divided into 2 groups according to the presence of ovarian endometriosis: group 1, no coexisting ovarian endometriosis; group 2, clear cell carcinoma arising from ovarian endometriosis or the presence of ovarian endometriosis elsewhere in the ovary. Clinicopathologic characteristics, disease-free survival (DFS), and overall survival (OS) were compared between the 2 groups. Of 155 patients with OCCC, 77 were categorized into group 1 and 78 into group 2. Group 2 patients were younger than group 1 (median age, 48 vs 51 years; P = 0.005) and had higher incidence of early-stage disease (stage I, 77% vs 58%; P = 0.001) and lower incidence of lymph node metastasis (4% vs 17%; P = 0.008). Group 2 patients were observed to have a significantly higher 5-year DFS (P < 0.001) and OS (P = 0.001) compared with group 1. In stage I disease, group 2 had a significantly higher 5-year DFS (P = 0.004) and OS (P = 0.016) than did group 1. In the multivariate analysis, coexisting endometriosis and advanced International Federation of Obstetrics and Gynecology stage were significant factors for both DFS and OS rates. Ovarian clear cell carcinoma with endometriosis was found more frequently in younger women and had a higher incidence of early-stage disease and a lower incidence of lymph node metastasis compared with OCCC without endometriosis. Ovarian endometriosis was associated with improved prognostic factors and a better DFS and OS even in stage I disease. Ovarian endometriosis was an independent prognostic factor for OCCC.


Sedentary behaviors and the risk of incident hypertension: the SUN Cohort.
Scientific evidence from different sources suggests a positive association between sedentary behaviors and the incidence of hypertension. However, no previous prospective study corroborated this potential relationship in an adult population. Our objective was to assess prospectively the association of different sedentary behaviors (interactive and noninteractive sedentary activity, television viewing, and sleeping) with the incidence of hypertension. In this prospective, dynamic cohort study (the SUN Study), 11,837 Spanish university graduates, with a mean age of 36 years, were followed for an average of 40 months, from 1999 to 2006. Sedentary behavior was assessed with a questionnaire at baseline, and the incidence of hypertension was assessed with validated, biennial questionnaires. We identified 291 new cases of hypertension among 6742 participants who remained after excluding those with self-reported hypertension, cardiovascular disease, diabetes, and cancer at baseline. Self-reported total sedentary behavior was directly associated with a higher risk of hypertension (hazard ratio, 1.48; 95% confidence interval, 1.01 to 2.18, comparing those in the upper and lower quartiles; P for trend = .03). In subtype analyses, interactive sedentary behavior (driving and computer use), but not noninteractive sedentary behavior (television viewing and sleeping), was associated with a higher risk of hypertension. Interactive sedentary behaviors appear to be an independent risk factor for incident hypertension. Noninteractive sedentary behaviors (television viewing and sleeping) did not show a significant association with incident hypertension.


Visit-to-Visit Variability in Total Cholesterol Correlates with the Progression of Renal Function Decline in a Chinese Community-Based Hypertensive Population.
Studies have demonstrated that cholesterol variability is an independent predictor of cerebrovascular and cardiovascular events. This study aimed to investigate the association of visit-to-visit variability in total cholesterol (TC) with kidney decline in a Chinese community-based population. We assessed intraindividual variability in TC among 6,465 hypertensive participants and correlated the results with endpoints. TC variability was measured using standard deviation (SD), average successive variability (ASV), coefficient of variation (CV), and variability independent of the mean (VIM). The endpoint of this study was progression of renal function decline defined as a decrease in estimated glomerular filtration rate (eGFR) ≥30% and to a level <60 mL/min/1.73 m2 during follow-up if the baseline eGFR was ≥60 mL/min/1.73 m2, or a decrease in eGFR ≥50% during follow up if the baseline eGFR was <60 mL/min/1.73 m2. After a median follow-up of 27 months, 13.5% (n = 877) of the participants experienced progression of renal function decline. In the multivariable-adjusted Cox model, each 1-SD increase in TC variability (by SD) increased the risk of renal function decline by 11% (HR = 1.11; 95% CI 1.034-1.197; p = 0.004); this was independent of the baseline eGFR, mean follow-up TC levels, and the lipid-lowering therapy. Similar results were found for the 3 other measures of variability, i.e., ASV, CV, and VIM. In subjects with hypertension, visit-to-visit variability in TC is an independent predictor of renal function decline.


Are persons with cognitive impairment able to state consistent choices?
This study examined the decision-making capacity of persons with cognitive impairment with respect to their everyday care preferences and choices. This is the first in a series of articles to report on findings from a larger study that examines choice, decision making, values, preferences, and practices in everyday care for community-dwelling persons with cognitive impairment and their family caregivers. Fifty-one respondent pairs, or dyads, were interviewed, that is, persons with cognitive impairment (n = 51) and their family caregivers (n = 51). All persons with cognitive impairment were interviewed twice within a week using a parallel interview to determine stability and accuracy of responses. The family caregiver was interviewed once. Persons with mild to moderate cognitive impairment (i.e., Mini-Mental State Exam scores 13-26) are able to respond consistently to questions about preferences, choices, and their own involvement in decisions about daily living, and to provide accurate and reliable responses to questions about demographics. Including the perspective of persons with cognitive impairment in both research and practice has the potential to enhance their autonomy and improve their quality of life.


Infectious complications and conservative treatment of totally implantable venous access devices in children with cancer.
Besides their complications, totally implantable venous access devices (TIVADs) increase the quality of life in children with cancer. The aim of this study was analysis of infectious complications and results of conservative management in TIVADs. Three hundred and one catheters were implanted in 283 patients between February 1991 and January 2005. Infectious complications were analyzed retrospectively. Cumulative duration of implantation was 153,757 days. In 140 devices (46.5%), no complication was detected. Total rate of infection was 1.96/1000 catheter days. Types of infections were as follows: catheterrelated bloodstream infections: 190; catheter-related systemic infections: 74; pocket infections: 19, exit site infections: 14; and tunnel infections: 5. Staphylococcus epidermidis and non-albicans candida were the most common isolations. During follow-up, a total of 119 catheters had been removed. Most of them were due to infection (n=42). In conclusion, TIVADs are important in children with cancer who need prolonged intravenous access, so they should be used carefully and managed conservatively in case of complications.


Xanthones of Lichen Source: A 2016 Update.
An update of xanthones encountered in lichens is proposed as more than 20 new xanthones have been described since the publication of the compendium of lichen metabolites by Huneck and Yoshimura in 1996. The last decades witnessed major advances regarding the elucidation of biosynthetic schemes leading to these fascinating compounds, accounting for the unique substitution patterns of a very vast majority of lichen xanthones. Besides a comprehensive analysis of the structures of xanthones described in lichens, their bioactivities and the emerging analytical strategies used to pinpoint them within lichens are presented here together with physico-chemical properties (including NMR data) as reported since 1996.


Contribution of sensorimotor integration to spinal stabilization in humans.
The control of upper body (UB) orientation relative to the pelvis in the frontal plane was characterized by analyzing responses to external perturbations consisting of continuous pelvis tilts (eyes open [EO] and eyes closed [EC]) and visual surround tilts (EO) at various amplitudes. Lateral sway of the lower body was prevented on all tests. UB sway was analyzed by calculating impulse-response functions (IRFs) and frequency-response functions (FRFs) from 0.023 to 10.3 Hz for pelvis tilt tests and FRFs from 0.041 to 1.5 Hz for visual tests. For pelvis tilt tests, differences between FRFs were limited to frequencies<3 Hz and were dependent on stimulus amplitude. IRFs were nearly identical across all pelvis tilt tests for the first 0.2 s, but showed amplitude-dependent changes in their time course at longer time lags. The availability of visual orientation cues (EO vs. EC) had only a small effect on the UB sway during pelvis tilt tests. This small effect of vision was consistent with the small UB sway evoked on visual tilt tests. Experimental results were interpreted using a feedback model of UB orientation control that included time-delayed sensory integration, short-latency reflexive mechanisms, and intrinsic biomechanical properties of the UB. Variation in model parameters indicated that subjects shifted toward reliance on vestibular information and away from proprioceptive information as pelvis tilt amplitudes increased. For visual tilt stimuli, model parameters indicated that subjects shifted toward reliance on vestibular and proprioceptive information and away from visual information as the stimulus amplitude increased.


Heritability and biochemistry of gangliosidosis in emus (Dromaius novaehollandiae).
The progeny of two emu breeder pairs, which had a history of producing offspring with gangliosidosis, were monitored for 15 mo. DNA fingerprinting revealed that individuals in each breeder pair were not related to each other. One breeder pair had 13 progeny that reached or exceeded the age of 1 mo, and six of these progeny developed gangliosidosis. The mean age at which these affected emus were euthanatized, with distinct neurologic disease, or died was 5.7 mo. The second emu pair had 13 progeny, seven of which developed gangliosidosis, with a mean age of euthanasia/death of 4.6 mo. Affected emus died or were euthanatized from 2 to 8 mo of age. The primary clinical sign in the affected emus was mild to severe ataxia. Severe hemorrhage into the body cavity or the muscles of the thigh was noted in 8 of 13 of the affected emus. Brain ganglioside levels were evaluated in six of the affected emus and six controls. Significant increases (P < 0.05) in gangliosides GM1 and GM3 were noted, with 2.3- and 4.9-fold increases in these two gangliosides, respectively, in affected emus. Furthermore, the diseased emu brains contained ganglioside GM2, whereas this monosialoganglioside was undetectable in the brains of normal controls. Total mean brain ganglioside sialic acid in affected emus was increased 3.3-fold in comparison with controls. Serum chemistries revealed elevated cholesterol and decreased uric acid levels in affected emus. Gangliosidosis in emus is an inherited disease process that, in the current study, caused 50% mortality in the progeny of two emu breeder pairs. The elimination of this lethal gene from emu breeder stock is essential for the long-term economic viability of the United States emu industry.


Screening extracts of Zanthoxylum chalybeum and Warburgia ugandensis for activity against measles virus (Swartz and Edmonston strains) in vitro.
A large proportion of the population in Uganda still relies on the use of plant extracts for treatment of various ailments. This study tested the claimed efficacy of some plants in the treatment of measles. In vitro antiviral assays were performed on extracts of two medicinal plants (Warburgia ugandensis and Zanthoxylum chalybeum) using measles virus (Edmonston and Swartz strains) as the test organisms. The assays performed were the neutralisation tests and the plaque reduction assays. Of the two plants Z. chalybeum had demonstrable in vitro antiviral activity in the seed extracts (titer reduction factor [TRF]: 100, for the ethanolic extract). The in vitro antiviral activity of the seed extracts was demonstrated to be due to compound 27-135D (TRF=1000), which was characterized by (1)H-NMR spectroscopy as the alkaloid skimmianine. Skimmianine had minimal toxicity to VERO cell lines. The petroleum ether extracts and the ethanolic extracts of Warburgia ugandensis had no inhibitory effect on cytopathic effect (CPE) formation, especially at the maximal non-toxic dose (MNTD). The extracts of W. ugandensis were highly toxic to VERO cell lines. The TRF values for the stem bark extracts of W. ugandensis were: water extract, 10; ethanolic extract, 1; fraction 27-163D, 100., which were regarded to be too low. Seed extracts of Z.chalybeum therefore probably cure measles due to the antiviral effect of skimmianine. It is not clear how extracts of W. ugandensis produce a beneficial response in measles disease, if at all.


Winner's Curse Correction and Variable Thresholding Improve Performance of Polygenic Risk Modeling Based on Genome-Wide Association Study Summary-Level Data.
Recent heritability analyses have indicated that genome-wide association studies (GWAS) have the potential to improve genetic risk prediction for complex diseases based on polygenic risk score (PRS), a simple modelling technique that can be implemented using summary-level data from the discovery samples. We herein propose modifications to improve the performance of PRS. We introduce threshold-dependent winner's-curse adjustments for marginal association coefficients that are used to weight the single-nucleotide polymorphisms (SNPs) in PRS. Further, as a way to incorporate external functional/annotation knowledge that could identify subsets of SNPs highly enriched for associations, we propose variable thresholds for SNPs selection. We applied our methods to GWAS summary-level data of 14 complex diseases. Across all diseases, a simple winner's curse correction uniformly led to enhancement of performance of the models, whereas incorporation of functional SNPs was beneficial only for selected diseases. Compared to the standard PRS algorithm, the proposed methods in combination led to notable gain in efficiency (25-50% increase in the prediction R2) for 5 of 14 diseases. As an example, for GWAS of type 2 diabetes, winner's curse correction improved prediction R2 from 2.29% based on the standard PRS to 3.10% (P = 0.0017) and incorporating functional annotation data further improved R2 to 3.53% (P = 2×10-5). Our simulation studies illustrate why differential treatment of certain categories of functional SNPs, even when shown to be highly enriched for GWAS-heritability, does not lead to proportionate improvement in genetic risk-prediction because of non-uniform linkage disequilibrium structure.


Associations of body mass index, physical activity and sedentary time with blood pressure in primary school children from south-west England: A prospective study.
Elevated blood pressure in children is a significant risk factor for the development of cardiovascular disease in adulthood. We examined how children's body mass index (BMI), physical activity and sedentary time at ages 9 and 11 are associated with blood pressure at age 11. Data were from 1283 children from Bristol, UK, who participated in the study aged 11 years, 797 of whom also participated in the study aged 9 years. Child height, weight and blood pressure were measured, and children wore accelerometers for five days, from which moderate-to-vigorous-intensity physical activity and sedentary minutes per day were derived. Multiple imputation of missing data and adjusted linear and logistic regression models were used to examine associations. Child BMI at 11 years was cross-sectionally associated with higher systolic and diastolic blood pressure (mean difference [95% confidence interval]: 0.91 [0.32 to 1.50] mm Hg and 1.08 [0.54 to 1.62] mm Hg, respectively, per standard deviation (SD) of BMI). BMI at age 9 was also positively associated with diastolic blood pressure at age 11 (1.16 mmHg per two years [0.49 to 1.84], per SD of BMI). For girls, sedentary time at age 9 years was associated with increased odds of having high systolic blood pressure at age 11 (odds ratio: 1.08 [1.01 to 1.16], per 10 minutes per day). There was no evidence of associations between sedentary time and blood pressure among boys. Similarly, there was little evidence that physical activity was associated with blood pressure in either cross-sectional or prospective analyses. Effective strategies are needed to prevent excess bodyweight among children in order to reduce cardiovascular disease risk.


Three linked enzyme loci in fishes: implications in the evolution of vertebrate chromosomes.
A three-point linkage group comprised of loci coding for adenosine deaminase (ADA), glucose-6-phosphate dehydrogenase (G6PDH), and 6-phospho-gluconate dehydrogenase (6PGD) is described in fish of the genus Xiphophorus (Poeciliidae). The alleles at loci in this group were shown to assort independently from the alleles at three other loci--isocitrate dehydrogenase 1 and 2, and glyceraldehyde-3-phosphate dehydrogenase 1. Alleles at the latter three loci also assort independently from each other. Data were obtained by observing the segregation of electrophoretically variant alleles in reciprocal backcross hybrids derived from crosses between either X. helleri guentheri or X. h. strigatus and X. maculatus. The linkage component of chi2 was significant (less than 0.01) in all crosses, indicating that the linkage group is conserved in all populations of both species of Xiphophorus examined. While data from X. h. guentheri backcrosses indicate the linkage relationship ADA--6%--G6PDH--24%--6PGD, and ADA--29%--6PGD (30% when corrected for double crossovers), data from backcrosses involving strigatus, while supporting the same gene order, yielded significantly different recombination frequencies. The likelihood of the difference being due to an inversion could not be separated from the possibility of a sex effect on recombination in the present data. The linkage of 6PGD and G6PDH has been shown to exist in species of at least three classes of vertebrates, indicating the possibility of evolutionary conservation of this linkage.


The demonstration of the succinic dehydrogenase system in Bacillus subtilis using tetranitro--blue tetrazolium combined with techniques of electron microscopy.
Activity of the succinic dehydrogenase system was studied in Bacillus subtilis utilizing combined techniques of cytochemistry and electron microscopy. Organisms were incubated in a medium containing tetranitro-blue tetrazolium (TNBT) which served as an electron acceptor. Enzymatic activity, as evidenced by deposition of TNBT-formazan, was found on membranous organelles associated with the cytoplasmic membrane and septal plasma membrane, the nuclear area, and the plasma membrane. Flagella, approximately 190 A in diameter, with thorn-like projections protruded through the cell wall. Tangential-oblique sections of the cell wall showed many pores approximately 220 A in diameter with a center-to-center spacing of approximately 450 A.


Inhibitory Effects of the Extracts of Juglans sigillata Green Husks on the Proliferation, Migration and Survival of KYSE150 and EC9706 Human Esophageal Cancer Cell Lines.
This study evaluated the antitumor activity of the extracts of green husks of Juglans sigillata Dode on esophageal cancer. KYSE150 EC9706 cells were treated with different concentrations of six components of the extracts of J. sigillata green husks. Cell viability was measured by MTT. Cell migration and cell invasion were measured by wound-healing assays and transwell assays, respectively. Cell apoptosis and cycle were measured by flow cytometry. The expression of cell migration, cell cycle and cell apoptosis regulatory proteins was analyzed by Western blotting. Only the three constituents, including EtOH extractives, EtOAc soluble fraction and gallic acid (GA), exhibited inhibitory effects on the cell viability, migration and invasion by decreasing MMP2 and MMP9 expression (all P < 0.05). Flow cytometry revealed that these three constituents also induced cell apoptosis by increasing Bax and cleaved caspase-3 but decreasing Bcl-2 in KYSE150 and EC9706 cells. Furthermore, these constituents arrested the cell cycle at G0/G1 by downregulating the expression of Cyclin D1 but upregulating p53 and phospho-p53 expression in KYSE150 cells. In conclusion, the green husks of J. sigillata may act as a potential inhibitor on esophageal cancer growth. GA was the major single active constituent of the extracts.


Characterisation of HIV-1 transmission clusters and drug-resistant mutations in Denmark, 2004 to 2016.
This study describes the prevalence of human immunodeficiency virus (HIV) drug resistance mutations among 1,815 patients in Denmark from 2004 to 2016 and characterises transmission clusters. POL sequences were analysed for subtype, drug resistance mutations and phylogenetic relationship. The prevalence of surveillance drug resistance mutations (SDRM) was 6.7%, while the prevalence of drug resistance mutations (DRM) with a clinical impact was 12.3%. We identified 197 transmission clusters with 706 patients. Patients 40 years or older were less likely to be members of a transmission cluster and patients in transmission clusters were less likely to be infected abroad. The proportion of late presenters (LP) was lower in active compared with inactive clusters. Large active clusters consisted of more men who have sex with men (MSM), had members more frequently infected in Denmark and contained a significantly lower proportion of LP and significantly fewer patients with DRM than small active clusters. Subtyping demonstrated that the Danish HIV epidemic is gradually becoming more composed of non-B subtypes/circulating recombinant forms. This study shows that active HIV-1 transmission has become increasingly MSM-dominated and that the recent increase in SDRM and DRM prevalence is not associated with more sustained transmission within identified transmission networks or clusters.


Microenvironmental protection of CML stem and progenitor cells from tyrosine kinase inhibitors through N-cadherin and Wnt-β-catenin signaling.
Tyrosine kinase inhibitors (TKIs) are highly effective in treatment of chronic myeloid leukemia (CML) but do not eliminate leukemia stem cells (LSCs), which remain a potential source of relapse. TKI treatment effectively inhibits BCR-ABL kinase activity in CML LSCs, suggesting that additional kinase-independent mechanisms contribute to LSC preservation. We investigated whether signals from the bone marrow (BM) microenvironment protect CML LSCs from TKI treatment. Coculture with human BM mesenchymal stromal cells (MSCs) significantly inhibited apoptosis and preserved CML stem/progenitor cells following TKI exposure, maintaining colony-forming ability and engraftment potential in immunodeficient mice. We found that the N-cadherin receptor plays an important role in MSC-mediated protection of CML progenitors from TKI. N-cadherin-mediated adhesion to MSCs was associated with increased cytoplasmic N-cadherin-β-catenin complex formation as well as enhanced β-catenin nuclear translocation and transcriptional activity. Increased exogenous Wnt-mediated β-catenin signaling played an important role in MSC-mediated protection of CML progenitors from TKI treatment. Our results reveal a close interplay between N-cadherin and the Wnt-β-catenin pathway in protecting CML LSCs during TKI treatment. Importantly, these results reveal novel mechanisms of resistance of CML LSCs to TKI treatment and suggest new targets for treatment designed to eradicate residual LSCs in CML patients.


Schistosoma mansoni: anti-SMw32 proteinase response in vaccinated and challenged baboons.
Antibodies to a cysteinyl proteinase of the trematode Schistosoma mansoni have been detected in serum from infected mice and humans. We have evaluated antiproteinase responses in infected baboons and in baboons vaccinated with irradiated, cryopreserved schistosomules prior to challenge. Prechallenge sera and normal, uninfected control sera were nonreactive by ELISA and immunoblots. Serum antibodies were first detectable by ELISA at two months post-challenge in both challenged (C) and vaccinated-challenged (V-C) baboons (serum dilution 1:200). By four months post-challenge, ELISA absorbance values for subgroup C baboons were significantly higher than for V-C counterparts. The immunoblot technique provided a more sensitive means of detecting antibody early in the infection. One month post-challenge, 7 of 12 C and V-C sera (diluted 1:100) contained measurable anti-proteinase antibody. By month two, 12 of 12 were immunoblot-positive. Baboons vaccinated but not challenged (subgroup V) remained negative. The presence of the anti-proteinase antibody appears to be a sensitive and early marker for infection by S. mansoni.


An unbiased in vitro screen for activating epidermal growth factor receptor mutations.
Cancer tissues harbor thousands of mutations, and a given oncogene may be mutated at hundreds of sites, yet only a few of these mutations have been functionally tested. Here, we describe an unbiased platform for the functional characterization of thousands of variants of a single receptor tyrosine kinase (RTK) gene in a single assay. Our in vitroscreen for activating mutations (iSCREAM) platform enabled rapid analysis of mutations conferring gain-of-function RTK activity promoting clonal growth. The screening strategy included a somatic model of cancer evolution and utilized a library of 7,216 randomly mutated epidermal growth factor receptor (EGFR) single-nucleotide variants that were tested in murine lymphoid Ba/F3 cells. These cells depend on exogenous interleukin-3 (IL-3) for growth, but this dependence can be compensated by ectopic EGFR overexpression, enabling selection for gain-of-function EGFR mutants. Analysis of the enriched mutants revealed EGFR A702V, a novel activating variant that structurally stabilized the EGFR kinase dimer interface and conferred sensitivity to kinase inhibition by afatinib. As proof of concept for our approach, we recapitulated clinical observations and identified the EGFR L858R as the major enriched EGFR variant. Altogether, iSCREAM enabled robust enrichment of 21 variants from a total of 7,216 EGFR mutations. These findings indicate the power of this screening platform for unbiased identification of activating RTK variants that are enriched under selection pressure in a model of cancer heterogeneity and evolution.


Evidence That IAA Conjugates Are Slow-Release Sources of Free IAA in Plant Tissues.
Evidence that indoleacetic acid (IAA) conjugates are metabolized via enzyme-catalyzed hydrolysis to free IAA and that their biological activities are related to the rates at which they are hydrolyzed by the tissue is presented. These conclusions are based on the following observations. Slow but continuous decarboxylation of the IAA moiety of IAA-l-alanine and IAA-glycine occurs when these conjugates are applied to pea (Pisum sativum L. cv. Alaska) stem segments. Inasmuch as IAA conjugates are protected from peroxidase-catalyzed oxidative decarboxylation, the conjugates are probably hydrolyzed and the freed IAA then further metabolized. Free IAA and IAA-l-alanine are converted, by pea stem tissue, into the same metabolites. The metabolism is enzymic, since conjugates of IAA with the d-isomers of the amino acids are inactive. Ethylene production induced by IAA-l-alanine and by IAA-glycine is correlated with their hydrolysis, as indicated by their decarboxylation and with the appearance or nonappearance of IAA metabolites in the tissues.


Treatment of Sjögren's syndrome-associated dry eye an evidence-based review.
Outcomes-based review of reported treatment options for patients with dry eye secondary to Sjögren's syndrome (SS). Dry eye affects many individuals worldwide. Significant proportion of patients with dry eye has underlying SS, a progressive autoimmune condition. The few suggested guidelines for the treatment of dry eye are mostly based on severity of symptoms and/or clinical findings rather than on outcomes analysis, and do not differentiate SS from other causes of dry eye. METHODS AND LITERATURE REVIEW: A search strategy was developed to identify prospective, interventional studies of treatments for SS-associated dry eye from electronic databases. Eligible references were restricted to English-language articles published after 1975. These sources were augmented by hand searches of reference lists from accessed articles. Study selection, data extraction, and grading of evidence were completed independently by ≥4 review authors. The searches identified 3559 references as of August 10, 2010. After duplicate review of the titles and abstracts, 245 full-text papers were assessed, 62 of which were relevant for inclusion in the review. In the current literature on SS-associated dry eye, there is a paucity of rigorous clinical trials to support therapy recommendations. Nonetheless, the recommended treatments include topical lubricants, topical anti-inflammatory therapy, and tear-conserving strategies. The efficacy of oral secretagogues seems greater in the treatment of oral dryness than ocular dryness. Although oral hydroxychloroquine is commonly prescribed to patients with SS to alleviate fatigue and arthralgias, the literature lacks strong evidence for the efficacy of this treatment for dry eye.


Combination of selected enzymes with cetyltrimethylammonium bromide in biofilm inactivation, removal and regrowth.
Enzymes are considered an innovative and environmentally friendly approach for biofilm control due to their lytic and dispersal activities. In this study, four enzymes (β-glucanase, α-amylase, lipase and protease) were tested separately and in combination with the quaternary ammonium compound cetyltrimethylammonium bromide (CTAB) to control flow-generated biofilms of Pseudomonas fluorescens. The four enzymes caused modest reduction of biofilm colony forming units (CFU). Protease, β-glucanase and α-amylase also caused modest biofilm removal. CTAB combined with either β-glucanase or α-amylase increased biofilm removal. Its combination with either β-glucanase or protease increased CFU reduction. However, CTAB-protease combination was antagonist in biofilm removal. Long-term effects in biofilm mass reduction were observed after protease exposure. In contrast, biofilms treated with β-glucanase were able to regrow significantly after exposure. Moreover, short-term respirometry tests with planktonic cells were performed to understand the effects of enzymes and their combination with CTAB on P. fluorescens viability. Protease and lipase demonstrated antimicrobial action, while α-amylase increased bacterial metabolic activity. The combination of CTAB with either protease or α-amylase was antagonistic, decreasing the antimicrobial action of CTAB. The overall results demonstrate a modest effect of the selected enzymes in biofilm control, either when applied alone or each one in combination with CTAB. Total biofilm removal or CFU reduction was not achieved and, in some cases, the use of enzymes antagonized the effects of CTAB. The results also propose that complementary tests, to characterize biofilm integrity and microbial viability, are required when someone is trying to assess the role of novel biocide - enzyme mixtures for effective biofilm control.


Impaired leukocyte-endothelial cell interaction in spontaneously hypertensive rats.
Hypertension is associated with a progressive organ injury whose etiology remains largely speculative. An increasing database shows that activated leukocytes, while affording an important immune protection, may be a contributing factor to several of the pathogenetic features of the hypertension syndrome. The purpose of this study was to determine the extent to which the glucocorticoid pathway may be involved in the atypical kinetics of leukocytes in spontaneously hypertensive rats (SHR) compared with normotensive Wistar-Kyoto (WKY) rats. The typical venular leukocyte adhesion induced by histamine application was significantly lower in SHR, and a comparison of normalized leukocyte rolling velocity (VWBC/VRBC) showed the values to be significantly higher in SHR relative to WKY controls. This abnormal trend in adherent leukocyte numbers and in VWBC/VRBC values could be counteracted when SHR were pretreated with RU 486, a synthetic glucocorticoid inhibitor, and restored to the levels observed in WKY rats. Anti-P-selectin monoclonal antibody (PB1.3) attenuated in SHR and WKY rats the increment of adherent leukocyte numbers as well as the decrement of VWBC/VRBC value that developed under combined histamine and RU 486 superfusion. Furthermore, an anti-intercellular adhesion molecule-1 monoclonal antibody (1A29) served to attenuate the increment of adherent leukocyte number induced by a combination of histamine and RU 486 superfusion in WKY rats and SHR. The results indicate that the deficient leukocyte-endothelial cell interaction in SHR can be circumvented by a glucocorticoid inhibitor.


Emergency remote teaching in higher education: mapping the first global online semester.
Due to the Covid-19 pandemic that spread globally in 2020, higher education courses were subsequently offered in fully remote, online formats. A plethora of primary studies began investigating a range of topics exploring teaching and learning in higher education, particularly during the initial semester. In order to provide an overview and initial understanding of this emerging research field, a systematic mapping review was conducted that collates and describes the characteristics of 282 primary empirical studies. Findings reveal that research was carried out mostly descriptively and cross-sectionally, focusing predominantly on undergraduate students and their perceptions of teaching and learning during the pandemic. Studies originate from a broad range of countries, are overwhelmingly published open access, and largely focused on the fields of Health & Welfare and Natural Sciences, Mathematics & Statistics. Educational technology used for emergency remote teaching are most often synchronous collaborative tools, used in combination with text-based tools. The findings are discussed against pre-pandemic research on educational technology use in higher education teaching and learning, and perspectives for further research are provided. The online version contains supplementary material available at 10.1186/s41239-021-00282-x.


Effect of buspirone on withdrawal symptoms associated with smoking cessation.
Preliminary information suggests that buspirone hydrochloride ameliorates symptoms of nicotine withdrawal. In a double-blind, randomized, placebo-controlled trial, we determined the effects of buspirone on the withdrawal symptoms associated with smoking cessation in 40 long-term cigarette smokers. Subjects were randomized to 4 weeks of treatment with either buspirone (n = 20) or placebo (n = 20). Subjects were instructed to maintain their usual cigarette intake during the first 21 days of treatment and to cease smoking on day 22 of treatment. Withdrawal symptoms were subjectively rated before and during smoking cessation. Before smoking cessation, there were no significant differences in the intensity of any withdrawal symptom between the buspirone- and placebo-treated subjects, with the exception of a significantly higher rating for drowsiness in the buspirone group. During the smoking cessation period, the ratings for craving, anxiety, irritability, restlessness, and sadness were significantly lower in the buspirone group than the placebo group. There were no significant differences between the groups in their ratings for hunger, inability to concentrate, or drowsiness. Fifteen buspirone-treated subjects and nine placebo-treated subjects were able to abstain from cigarettes during the entire 7-day smoking cessation period. No subject dropped out of the study for perceived drug side effects. Eight buspirone-treated subjects and five placebo-treated subjects reported side effects, all of which were mild. Buspirone ameliorated most of the short-term withdrawal symptoms associated with smoking cessation. Further controlled studies will be needed to define the benefit in smoking cessation.


Moderation of phenotypic severity in dystrophic and junctional forms of epidermolysis bullosa through in-frame skipping of exons containing non-sense or frameshift mutations.
Non-sense mutations on both alleles of either the type VII collagen gene (COL7A1) or the genes encoding laminin 5 (LAMA3, LAMB3, or LAMC2) usually result in clinically severe forms of recessive dystrophic or junctional epidermolysis bullosa, respectively. In this study we assessed two unrelated families whose mutations in genomic DNA predicted severe recessive dystrophic epidermolysis bullosa or junctional epidermolysis bullosa phenotypes but in whom the manifestations were milder than expected. The recessive dystrophic epidermolysis bullosa patients had a homozygous single base-pair frameshift mutation in exon 19 of COL7A1 (2470insG). Clinically, there was generalized blistering but only mild scarring. Skin biopsy revealed positive type VII collagen immunoreactivity and recognizable anchoring fibrils. The junctional epidermolysis bullosa patients were compound heterozygotes for a frameshift/non-sense combination of mutations in exons 3 and 17 of LAMB3 (29insC/Q834X). These patients did not have the lethal form of junctional epidermolysis bullosa but, as adults, displayed the milder generalized atrophic benign epidermolysis bullosa variant. There was undetectable laminin 5 staining at the dermal-epidermal junction using an antibody to the beta3 chain, but faintly positive alpha3 and gamma2 chain labeling, and there was variable hypoplasia of hemidesmosomes. To explain the milder recessive dystrophic epidermolysis bullosa and junctional epidermolysis bullosa phenotypes in these families, reverse transcription-polymerase chain reaction, using RNA extracted from frozen skin, was able to provide evidence for some rescue of mutant mRNA transcripts with restoration of the open- reading frame. In the recessive dystrophic epidermolysis bullosa patients, transcripts containing in-frame skipping of exon 19 of COL7A1 in the cDNA were detected, and in the junctional epidermolysis bullosa patients transcripts with in-frame skipping of exon 17 of LAMB3 were identified. The truncated proteins encoded by these transcripts are expected to lack certain critical domains involved in cell-matrix attachment, but may still be able to contribute to adhesion thereby moderating the severity of the skin blistering. This study shows the limitations in predicting phenotype in epidermolysis bullosa solely based on mutation analysis of genomic DNA and emphasizes the importance of immunohistochemistry, electron microscopy, and mRNA assessment as parallel investigations.


Defensive inking in Aplysia spp: multiple episodes of ink secretion and the adaptive use of a limited chemical resource.
The seahare Aplysia spp. extracts many of its defensive chemicals from its red seaweed diet, including its purple ink, which is an effective deterrent against predators such as anemones and crabs. It is believed that the inking behavior is a high-threshold, all-or-none fixed act that nearly completely depletes the seahare of its ink supply. If a seahare depletes its gland of ink, it must seek out a source of red seaweed and then feed for at least 2 days to replenish its ink supply. This suggests that the animal would not be able to deploy ink more than once in rapid succession in response to successive attacks from one or more predators. However, we found that Aplysia spp. can secrete ink in response to three or more successive stimulations with (i) anemone tentacles, (ii) a mechanical stimulus, consisting of grabbing and lifting the animal from the substratum, or (iii) a noxious electric shock. A spectro-photometric measure of ink secretion showed that only approximately 48 % of the gland's releasable ink reserves are deployed initially. Thus, deployment of this defensive chemical is not strictly all-or-nothing, although the trigger mechanism is. Moreover, the animal tends to secrete a relatively fixed proportion (30--50 %) of its available ink reserves even after its gland has been depleted to approximately half its initial content. Since an animal need only use a proportion of its ink reserves to deter an attacker effectively, the inking behavior is adaptive in its economical use of a limited resource.


Bound exciton photoluminescence from ion‑implanted phosphorus in thin silicon layers.
We report the observation of clear bound exciton (BE) emission from ion-implanted phosphorus. Shallow implantation and high-temperature annealing successfully introduce active donors into thin silicon layers. The BE emission at a wavelength of 1079 nm shows that a part of the implanted donors are definitely activated and isolated from each other. However, photoluminescence and electron spin resonance studies find a cluster state of the activated donors. The BE emission is suppressed by this cluster state rather than the nonradiative processes caused by ion implantation. Our results provide important information about ion implantation for doping quantum devices with phosphorus quantum bits.


A world-wide analysis of reduced sensitivity to DMI fungicides in the banana pathogen Pseudocercospora fijiensis.
Pseudocercospora fijiensis is the causal agent of the black leaf streak disease (BLSD) of banana. Bananas are important global export commodities and a major staple food. Their susceptibility to BLSD pushes disease management towards excessive fungicide use, largely relying on multisite inhibitors and sterol demethylation inhibitors (DMIs). These fungicides are ubiquitous in plant disease control, targeting the CYP51 enzyme. We examined sensitivity to DMIs in P. fijiensis field isolates collected from various major banana production zones in Colombia, Costa Rica, Dominican Republic, Ecuador, the Philippines, Guadalupe, Martinique and Cameroon and determined the underlying genetic reasons for the observed phenotypes. We observed a continuous range of sensitivity towards the DMI fungicides difenoconazole, epoxiconazole and propiconazole with clear cross-sensitivity. Sequence analyses of PfCYP51 in 266 isolates showed 28 independent amino acid substitutions, nine of which correlated with reduced sensitivity to DMIs. In addition to the mutations, we observed up to six insertions in the Pfcyp51 promoter. Such promoter insertions contain repeated elements with a palindromic core and correlate with the enhanced expression of Pfcyp51 and hence with reduced DMI sensitivity. Wild-type isolates from unsprayed bananas fields did not contain any promoter insertions. The presented data significantly contribute to understanding of the evolution and global distribution of DMI resistance mechanisms in P. fijiensis field populations and facilitate the prediction of different DMI efficacy. The overall reduced DMI sensitivity calls for the deployment of a wider range of solutions for sustainable control of this major banana disease. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


The benefits of 'One Health' for pastoralists in Africa.
'One health' is particularly suited to serve mobile pastoralists. Dinka pastoralists in Sudan inspired Calvin Schwabe to coin the term 'one medicine', indicating that there is no difference in paradigm between human and veterinary medicine. Our contemporary definition of 'one health' is any added value in terms of improved health of humans and animals or financial savings or environmental services resulting from a closer cooperation of human and animal health sectors. Here we present a summary of 'one health' studies with mobile pastoralists in Africa which were done in research partnership, demonstrating such an added value. Initial joint human and animal health studies revealed higher livestock vaccination coverage than in the pastoralist community, leading to joint animal and human vaccination intervention studies which demonstrated a better access to primary health care services for pastoralists in Chad. Further simultaneous animal and human serological studies showed that camel breeding was associated with human Q-fever seropositivity. In Borana communities in Ethiopia, human cases of Mycobacterium bovis infection could be related to strains isolated from cattle. A challenge remained with regard to how to assess vaccination coverage in mobile populations. With the advent of mobile phones, health and demographic surveillance could be established for mobile pastoralists and their animals. This presents vast possibilities for surveillance and control of human and animal diseases. Pastoralists prefer a 'one health' approach and therefore contribute toward the validation of this concept by showing real added value of the cooperation between human and animal health services.


Memory Contextualization: The Role of Prefrontal Cortex in Functional Integration across Item and Context Representational Regions.
Memory recall is facilitated when retrieval occurs in the original encoding context. This context dependency effect likely results from the automatic binding of central elements of an experience with contextual features (i.e., memory "contextualization") during encoding. However, despite a vast body of research investigating the neural correlates of explicit associative memory, the neural interactions during encoding that predict implicit context-dependent memory remain unknown. Twenty-six participants underwent fMRI during encoding of salient stimuli (faces), which were overlaid onto unique background images (contexts). To index subsequent context-dependent memory, face recognition was tested either in intact or rearranged contexts, after scanning. Enhanced face recognition in intact relative to rearranged contexts evidenced successful memory contextualization. Overall subsequent memory effects (brain activity predicting whether items were later remembered vs. forgotten) were found in the left inferior frontal gyrus (IFG) and right amygdala. Effective connectivity analyses showed that stronger context-dependent memory was associated with stronger coupling of the left IFG with face- and place-responsive areas, both within and between participants. Our findings indicate an important role for the IFG in integrating information across widespread regions involved in the representation of salient items and contextual features.


Ewing sarcoma vs lymphoblastic lymphoma. A comparative immunohistochemical study.
To develop a practical immunohistochemistry panel for distinguishing lymphoblastic lymphoma from Ewing sarcoma (ES), we evaluated 17 ES and 27 lymphoblastic lymphoma and leukemia cases with antibodies to CD99, terminal deoxynucleotidyl transferase (TdT), leukocyte common antigen (LCA), CD43, CD79a, CD20, CD3, vimentin, and neuron-specific enolase (NSE). Three cases were bone lymphomas, 2 initially misdiagnosed as ES. All cases were CD99+. All lymphomas and leukemias were TdT+ compared to none of the ESs. None of the ESs expressed other lymphocytic markers, which were inconsistently expressed in the lymphomas and leukemias: CD43, 33%; LCA, 30%; CD79a, 19%; CD3, 19%; and CD20, 7%. Of the ESs, 88% were vimentin positive compared with 23% of lymphomas and leukemias. Vimentin was stronger and more diffuse in ES. NSE did not reliably stain any cases. When faced with the differential diagnosis of ES vs lymphoblastic lymphoma, an immunohistochemical panel that includes antibodies to CD99 and TdT is useful. Both epitopes are well preserved in fixed and decalcified tissue. A panel composed of antibodies to CD99 and TdT, in conjunction with other lymphocytic markers and vimentin, is highly sensitive and specific.


Micropatterning of poly(ethylene glycol) diacrylate hydrogels.
This protocol describes the techniques to synthesize and fabricate micropatterned poly(ethylene glycol) diacrylate-based hydrogels that can be used as substrates in cellular studies and tissue engineering scaffolds. These materials provide an essentially bioinert background material due to the very low protein adsorption characteristics of poly(ethylene glycol), but the materials can be modified with covalently grafted peptides, proteins, or other biomolecules of interest to impart specific biofunctionality to the material. Further, it is possible to use micropatterning technologies to control the localization of such covalent grafting of biomolecules to the hydrogel materials, thus spatially controlling the cell-material interactions. This protocol presents a relatively simple approach for mask-based photolithographic patterning, generally best suited for patterning the surface of hydrogel materials for 2D cell studies. A more sophisticated technique, two-photon laser scanning lithography, is also presented. This technique allows free-form, 3D micropatterning in hydrogels.


The Spatial Dynamics of Predators and the Benefits and Costs of Sharing Information.
Predators of all kinds, be they lions hunting in the Serengeti or fishermen searching for their catch, display various collective strategies. A common strategy is to share information about the location of prey. However, depending on the spatial characteristics and mobility of predators and prey, information sharing can either improve or hinder individual success. Here, our goal is to investigate the interacting effects of space and information sharing on predation efficiency, represented by the expected rate at which prey are found and consumed. We derive a feeding functional response that accounts for both spatio-temporal heterogeneity and communication, and validate this mathematical analysis with a computational agent-based model. This agent-based model has an explicit yet minimal representation of space, as well as information sharing about the location of prey. The analytical model simplifies predator behavior into a few discrete states and one essential trade-off, between the individual benefit of acquiring information and the cost of creating spatial and temporal correlation between predators. Despite the absence of an explicit spatial dimension in these equations, they quantitatively predict the predator consumption rates measured in the agent-based simulations across the explored parameter space. Together, the mathematical analysis and agent-based simulations identify the conditions for when there is a benefit to sharing information, and also when there is a cost.


Phosphatidylinositol 4,5-bisphosphate induces actin-based movement of raft-enriched vesicles through WASP-Arp2/3.
Phosphatidylinositol 4,5-bisphosphate (PIP(2)) has been implicated in the regulation of the actin cytoskeleton and vesicle trafficking. It stimulates de novo actin polymerization by activating the pathway involving the Wiskott-Aldrich syndrome protein (WASP) and the actin-related protein complex Arp2/3. Other studies show that actin polymerizes from cholesterol-sphingolipid-rich membrane microdomains called 'rafts', in a manner dependent on tyrosine phosphorylation. Although actin has been implicated in vesicle trafficking, and rafts are sites of active phosphoinositide and tyrosine kinase signaling that mediate apically directed vesicle trafficking, it is not known whether phosphoinositide regulation of actin dynamics occurs in rafts, or if it is linked to vesicle movements. Overexpression of type I phosphatidylinositol phosphate 5-kinase (PIP5KI), which synthesizes PIP(2), promoted actin polymerization from membrane-bound vesicles to form motile actin comets. Pervanadate (PV), a tyrosine phosphatase inhibitor, induced comets even in the absence of PIP5KI overexpression. PV increased PIP(2) levels, suggesting that it induces comets by changing PIP(2) homeostasis and by increasing tyrosine phosphorylation. Platelet-derived growth factor (PDGF) enhanced PV-induced comet formation, and these stimuli together potentiated the PIP5KI effect. The vesicles at the heads of comets were enriched in PIP5KIs and tyrosine phosphoproteins. WASP-Arp2/3 involvement was established using dominant-negative WASP constructs. Endocytic and exocytic markers identified vesicles enriched in lipid rafts as preferential sites of comet generation. Extraction of cholesterol with methyl-beta-cyclodextrin reduced comets, establishing that rafts promote comet formation. Sphingolipid-cholesterol rafts are preferred platforms for membrane-linked actin polymerization. This is mediated by in situ PIP(2) synthesis and tyrosine kinase signaling through the WASP-Arp2/3 pathway. Actin comets may provide a novel mechanism for raft-dependent vesicle transport and apical membrane trafficking.


Solvent water interactions within the active site of the membrane type I matrix metalloproteinase.
Matrix metalloproteinases (MMP) are an important family of proteases which catalyze the degradation of extracellular matrix components. While the mechanism of peptide cleavage is well established, the process of enzyme regeneration, which represents the rate limiting step of the catalytic cycle, remains unresolved. This step involves the loss of the newly formed N-terminus (amine) and C-terminus (carboxylate) protein fragments from the site of catalysis coupled with the inclusion of one or more solvent waters. Here we report a novel crystal structure of membrane type I MMP (MT1-MMP or MMP-14), which includes a small peptide bound at the catalytic Zn site via its C-terminus. This structure models the initial product state formed immediately after peptide cleavage but before the final proton transfer to the bound amine; the amine is not present in our system and as such proton transfer cannot occur. Modeling of the protein, including earlier structural data of Bertini and coworkers [I. Bertini, et al., Angew. Chem., Int. Ed., 2006, 45, 7952-7955], suggests that the C-terminus of the peptide is positioned to form an H-bond network to the amine site, which is mediated by a single oxygen of the functionally important Glu240 residue, facilitating efficient proton transfer. Additional quantum chemical calculations complemented with magneto-optical and magnetic resonance spectroscopies clarify the role of two additional, non-catalytic first coordination sphere waters identified in the crystal structure. One of these auxiliary waters acts to stabilize key intermediates of the reaction, while the second is proposed to facilitate C-fragment release, triggered by protonation of the amine. Together these results complete the enzymatic cycle of MMPs and provide new design criteria for inhibitors with improved efficacy.