Anti-VEGF therapy reduces intestinal inflammation in Endoglin heterozygous mice subjected to experimental colitis.
Chronic intestinal inflammation is associated with pathological angiogenesis that further amplifies the inflammatory response. Vascular endothelial growth factor (VEGF), is a major angiogenic cytokine that has been implicated in chronic colitis and inflammatory bowel diseases. Endoglin (CD105), a transforming growth factor-β superfamily co-receptor expressed on endothelial and some myeloid cells, is a modulator of angiogenesis involved in wound healing and potentially in resolution of inflammation. We showed previously that Endoglin heterozygous (Eng (+/-)) mice subjected to dextran sodium sulfate developed severe colitis, abnormal colonic vessels and high tissue VEGF. We therefore tested in the current study if treatment with a monoclonal antibody to VEGF could ameliorate chronic colitis in Eng (+/-) mice. Tissue inflammation and microvessel density (MVD) were quantified on histological slides. Colonic wall thickness, microvascular hemodynamics and targeted MAdCAM-1(+) inflamed vessels were assessed in vivo by ultrasound. Mediators of angiogenesis and inflammation were measured by Milliplex and ELISA assays. Colitic Eng (+/-) mice showed an increase in intestinal inflammation, MVD, colonic wall thickness, microvascular hemodynamics and the number of MAdCAM-1(+) microvessels relative to colitic Eng (+/+) mice; these parameters were all attenuated by anti-VEGF treatment. Of all factors up-regulated in the inflamed gut, granulocyte colony-stimulating factor (G-CSF) and amphiregulin were further increased in colitic Eng (+/-) versus Eng (+/+) mice. Anti-VEGF therapy decreased tissue VEGF and inflammation-induced endoglin, IL-1β and G-CSF in colitic Eng (+/-) mice. Our results suggest that endoglin modulates intestinal angiogenic and inflammatory responses in colitis. Furthermore, contrast-enhanced ultrasound provides an excellent non-invasive imaging modality to monitor gut angiogenesis, inflammation and responses to anti-angiogenic treatment.


A Novel C-Terminal CIB2 (Calcium and Integrin Binding Protein 2) Mutation Associated with Non-Syndromic Hearing Loss in a Hispanic Family.
Hearing loss is a complex disorder caused by both genetic and environmental factors. Previously, mutations in CIB2 have been identified as a common cause of genetic hearing loss in Pakistani and Turkish populations. Here we report a novel (c.556C>T; p.(Arg186Trp)) transition mutation in the CIB2 gene identified through whole exome sequencing (WES) in a Caribbean Hispanic family with non-syndromic hearing loss. CIB2 belongs to the family of calcium-and integrin-binding (CIB) proteins. The carboxy-termini of CIB proteins are associated with calcium binding and intracellular signaling. The p.(Arg186Trp) mutation is localized within predicted type II PDZ binding ligand at the carboxy terminus. Our ex vivo studies revealed that the mutation did not alter the interactions of CIB2 with Whirlin, nor its targeting to the tips of hair cell stereocilia. However, we found that the mutation disrupts inhibition of ATP-induced Ca2+ responses by CIB2 in a heterologous expression system. Our findings support p.(Arg186Trp) mutation as a cause for hearing loss in this Hispanic family. In addition, it further highlights the necessity of the calcium binding property of CIB2 for normal hearing.


Bacteremia in infants with Salmonella enterocolitis.
In order to determine the incidence of Salmonella bacteremia in children with Salmonella enterocolitis and to predict the risk factors of bacteremia, we conducted a prospective study on 264 infants and children with Salmonella enterocolitis from January 1, 1988 to December 31, 1989. They were admitted to the Pediatric Department of Mackay Memorial Hospital with the chief problem of diarrhea. The rectal swab cultures were all positive for nontyphoidal Salmonella. Blood cultures were done immediately when Salmonella enterocolitis was confirmed. Clinical features including age, clinical symptoms, laboratory data, and serogroup were all recorded and analyzed to predict the risk factors of bacteremia in Salmonella enterocolitis. Twenty patients had bacteremia. Among them, 4 had meningitis simultaneously. There was one death, resulting in a 5% mortality in the bacteremia group. The overall rate of bacteremia was 7.6%. For those less than 3 months of age, it was 11.5%. We were unable to find any relationship between the bacteremia and the risk factors such as age, body temperature, clinical symptoms, white blood cell counts and differential counts. Interestingly, it was noted that Salmonella serogroup D1, a highly virulent and invasive Salmonella serogroup, was more closely related to the occurrence of bacteremia. It accounted for 40% of all cases of bacteremia. In conclusion, this study showed that the incidence of bacteremia in cases of Salmonella enterocolitis was 7.6% and the only risk factor of bacteremia was a culture of Salmonella serogroup D1. Therefore, in cases of Salmonella serogroup D1 enterocolitis, antibiotics treatment may be considerable.


Electrical microcurrent to prevent conditioning film and bacterial adhesion to urological stents.
Long-term catheters remain a significant clinical problem in urology due to the high rate of bacterial colonization, infection, and encrustation. Minutes after insertion of a catheter, depositions of host urinary components onto the catheter surface form a conditioning film actively supporting the bacterial adhesion process. We investigated the possibility of reducing or avoiding the buildup of these naturally forming conditioning films and of preventing bacterial adhesion by applying different current densities to platinum electrodes as a possible catheter coating material. In this model we employed a defined environment using artificial urine and Proteus mirabilis. The film formation and desorption was analyzed by highly mass sensitive quartz crystal microbalance and surface sensitive atomic force microscopy. Further, we performed bacterial staining to assess adherence, growth, and survival on the electrodes with different current densities. By applying alternating microcurrent densities on platinum electrodes, we could produce a self regenerative surface which actively removed the conditioning film and significantly reduced bacterial adherence, growth, and survival. The results of this study could easily be adapted to a catheter design for clinical use.


It's a matter of design-how pitfall trap design affects trap samples and possible predictions.
Pitfall traps are commonly used to assess ground dwelling arthropod communities. The effects of different pitfall trap designs on the trapping outcome are poorly investigated however they might affect conclusions drawn from pitfall trap data greatly. We tested four pitfall trap types which have been used in previous studies for their effectiveness: a simple type, a faster exchangeable type with an extended plastic rim plate and two types with guidance barriers (V- and X-shaped). About 20 traps were active for 10 weeks and emptied biweekly resulting in 100 trap samples. Pitfall traps with guidance barriers were up to five times more effective than simple pitfall traps and trap samples resulted in more similar assemblage approximations. Pitfall traps with extended plastic rim plates did not only perform poorly but also resulted in distinct carabid assemblages with less individuals of small species and a larger variation. Due to the obvious trait filtering and resulting altered assemblages, we suggest not to use pitfall traps with extended plastic rim plates. In comprehensive biodiversity inventories, a smaller number of pitfall traps with guidance barriers and a larger number of spatial replicates is of advantage, while due to comparability reasons, the use of simple pitfall traps will be recommended in most other cases.


Effects of aberrations in spatiotemporal focusing of ultrashort laser pulses.
Spatiotemporal focusing, or simultaneous spatial and temporal focusing (SSTF), has already been adopted for various applications in microscopy, photoactivation for biological studies, and laser fabrication. We investigate the effects of aberrations on focus formation in SSTF, in particular, the effects of phase aberrations related to low-order Zernike modes and a refractive index mismatch between the immersion medium and sample. By considering a line focus, we are able to draw direct comparison between the performance of SSTF and conventional spatial focusing (SF). Wide-field SSTF is also investigated and is found to be much more robust to aberrations than either line SSTF or SF. These results show the sensitivity of certain focusing methods to specific aberrations, and can inform on the necessity and benefit of aberration correction.


Riboneogenesis in yeast.
Glucose is catabolized in yeast via two fundamental routes, glycolysis and the oxidative pentose phosphate pathway, which produces NADPH and the essential nucleotide component ribose-5-phosphate. Here, we describe riboneogenesis, a thermodynamically driven pathway that converts glycolytic intermediates into ribose-5-phosphate without production of NADPH. Riboneogenesis begins with synthesis, by the combined action of transketolase and aldolase, of the seven-carbon bisphosphorylated sugar sedoheptulose-1,7-bisphosphate. In the pathway's committed step, sedoheptulose bisphosphate is hydrolyzed to sedoheptulose-7-phosphate by the enzyme sedoheptulose-1,7-bisphosphatase (SHB17), whose activity we identified based on metabolomic analysis of the corresponding knockout strain. The crystal structure of Shb17 in complex with sedoheptulose-1,7-bisphosphate reveals that the substrate binds in the closed furan form in the active site. Sedoheptulose-7-phosphate is ultimately converted by known enzymes of the nonoxidative pentose phosphate pathway to ribose-5-phosphate. Flux through SHB17 increases when ribose demand is high relative to demand for NADPH, including during ribosome biogenesis in metabolically synchronized yeast cells.


A comparative study of the prevalence of and factors associated with insecticide-treated nets usage among children under 5 years of age in households that already own nets in Malawi.
Despite malaria control programmes having successfully increased the number of households owning insecticide-treated nets (ITNs) in Malawi, the population of people with ITN access but still not using them fluctuated from 13% in 2010, 5% in 2012 and then 12% in 2015. This study aimed to compare the rate and factors associated with ITN usage among children under 5 years of age, living in household with at least one ITN, in Malawi between 2010 and 2015. The 2010 and 2015-2016 Malawi Demographic and Health Surveys (MDHSs) were utilized. Only children from households that owned at least one ITN were selected. Multivariate logistic regression analyses were performed to examine associations of child, maternal and household factors with ITN usage. In total, 12,378 and 10,196 children under 5 years of age were examined from 2010 and 2015-2016, respectively. ITN usage increased from 57.8% (95% Confidence interval (CI): 56.1%-59.4%) in 2010 to 69.0% (95% CI 67.4%-70.5%) in 2015. The multivariate analysis revealed that, among others, being aged ≥ 24 months, having mothers with no formal education or with primary education, residing in a female-headed household, and residing in households that had poor household ITN supply were significantly associated with reduced odds of ITN usage. ITNs are a key vector control intervention in malaria prevention. This study revealed increased ITN usage among children under 5 years old in the 5-year period, suggesting that considerable improvements have been made. However, continued efforts to increase awareness of the importance of using ITNs in malaria prevention in Malawi are necessary. Findings from this research provide some policy implications, especially for improving household ITN supply, to improve ITN utilization in Malawi.


Brain Magnetic Resonance Imaging Reveals Different Courses of Disease in Pediatric and Adult Cerebral Malaria.
Cerebral malaria is a common presentation of severe Plasmodium falciparum infection and remains an important cause of death in the tropics. Key aspects of its pathogenesis are still incompletely understood, but severe brain swelling identified by magnetic resonance imaging (MRI) was associated with a fatal outcome in African children. In contrast, neuroimaging investigations failed to identify cerebral features associated with fatality in Asian adults. Quantitative MRI with brain volume assessment and apparent diffusion coefficient (ADC) histogram analyses were performed for the first time in 65 patients with cerebral malaria to compare disease signatures between children and adults from the same cohort, as well as between fatal and nonfatal cases. We found an age-dependent decrease in brain swelling during acute cerebral malaria, and brain volumes did not differ between fatal and nonfatal cases across both age groups. In nonfatal disease, reversible, hypoxia-induced cytotoxic edema occurred predominantly in the white matter in children, and in the basal ganglia in adults. In fatal cases, quantitative ADC histogram analyses also demonstrated different end-stage patterns between adults and children: Severe hypoxia, evidenced by global ADC decrease and elevated plasma levels of lipocalin-2 and microRNA-150, was associated with a fatal outcome in adults. In fatal pediatric disease, our results corroborate an increase in brain volume, leading to augmented cerebral pressure, brainstem herniation, and death. Our findings suggest distinct pathogenic patterns in pediatric and adult cerebral malaria with a stronger cytotoxic component in adults, supporting the development of age-specific adjunct therapies.


Relationship of food groups and water intake to colon cancer risk.
The association between food groupings and adenocarcinoma of the colon was investigated in a population-based case-control study of men and women ages 30-62 years. Colon cancer cases (238 men and 186 women) diagnosed from 1985 to 1989 were identified from the Seattle-Puget Sound Surveillance, Epidemiology, and End Results Registry. Controls (224 men and 190 women) were selected using a random digit telephone dialing method. Dietary information was gathered using an 80-item food frequency questionnaire. Foods were grouped and analyzed by quartile of intake, with adjustment for age and total energy intake. Among women, a reduced risk of colon cancer was associated with a high intake of fruits and vegetables [adjusted odds ratio (OR) for highest versus lowest quartile, 0.48; 95% confidence interval (CI), 0.26-0.86; P for trend, P = 0.02]. Inverse associations were also observed for the consumption of total (hot and cold) cereals (OR, 0.47; 95% CI, 0.25-0.91; P = 0.05), dairy products (OR, 0.40; 95% CI, 0.21-0.79; P = 0.05), and water (OR for > 5 glasses/day versus < or = 2 glass/day, 0.55; 95% CI, 0.31-0.99; P = 0.004). Among men, colon cancer risk was inversely associated with the intake of breads and cereals (OR, 0.43; 95% CI, 0.22-0.82; P = 0.02) and hot cereal (OR for weekly versus never eating, 0.53; 95% CI, 0.32-0.87; P = 0.01). Water consumption was marginally associated with a decreased colon cancer risk among men as well (OR for > 4 glasses/day versus < or = 1 glass/day, 0.68; 95% CI, 0.38-1.22; P = 0.16). Total meat consumption was associated with an increased risk of distal colon cancer among men (OR, 2.20; 95% CI, 1.08-4.48; P = 0.01). These results were not confounded by body mass index or other measured health behaviors. Results of this research support previous findings which associate intake of fruits, vegetables, grains, and dairy products with reduced colon cancer risk, and meat intake with an increased colon cancer risk. This study also reports a new finding of a possible inverse association of water consumption (glasses of plain water per day) with colon cancer risk.


A large-scale chemical screen for regulators of the arginase 1 promoter identifies the soy isoflavone daidzeinas a clinically approved small molecule that can promote neuronal protection or regeneration via a cAMP-independent pathway.
An ideal therapeutic for stroke or spinal cord injury should promote survival and regeneration in the CNS. Arginase 1 (Arg1) has been shown to protect motor neurons from trophic factor deprivation and allow sensory neurons to overcome neurite outgrowth inhibition by myelin proteins. To identify small molecules that capture Arg1's protective and regenerative properties, we screened a hippocampal cell line stably expressing the proximal promoter region of the arginase 1 gene fused to a reporter gene against a library of compounds containing clinically approved drugs. This screen identified daidzein as a transcriptional inducer of Arg1. Both CNS and PNS neurons primed in vitro with daidzein overcame neurite outgrowth inhibition from myelin-associated glycoprotein, which was mirrored by acutely dissociated and cultured sensory neurons primed in vivo by intrathecal or subcutaneous daidzein infusion. Further, daidzein was effective in promoting axonal regeneration in vivo in an optic nerve crush model when given intraocularly without lens damage, or most importantly, when given subcutaneously after injury. Mechanistically, daidzein requires transcription and induction of Arg1 activity for its ability to overcome myelin inhibition. In contrast to canonical Arg1 activators, daidzein increases Arg1 without increasing CREB phosphorylation, suggesting its effects are cAMP-independent. Accordingly, it may circumvent known CNS side effects of some cAMP modulators. Indeed, daidzein appears to be safe as it has been widely consumed in soy products, crosses the blood-brain barrier, and is effective without pretreatment, making it an ideal candidate for development as a therapeutic for spinal cord injury or stroke.


E2F1 proteolysis via SCF-cyclin F underlies synthetic lethality between cyclin F loss and Chk1 inhibition.
Cyclins are central engines of cell cycle progression in conjunction with cyclin-dependent kinases (CDKs). Among the different cyclins controlling cell cycle progression, cyclin F does not partner with a CDK, but instead forms via its F-box domain an SCF (Skp1-Cul1-F-box)-type E3 ubiquitin ligase module. Although various substrates of cyclin F have been identified, the vulnerabilities of cells lacking cyclin F are not known. Thus, we assessed viability of cells lacking cyclin F upon challenging them with more than 180 different kinase inhibitors. The screen revealed a striking synthetic lethality between Chk1 inhibition and cyclin F loss. Chk1 inhibition in cells lacking cyclin F leads to DNA replication catastrophe. Replication catastrophe depends on accumulation of the transcription factor E2F1 in cyclin F-depleted cells. We find that SCF-cyclin F controls E2F1 ubiquitylation and degradation during the G2/M phase of the cell cycle and upon challenging cells with Chk1 inhibitors. Thus, Cyclin F restricts E2F1 activity during the cell cycle and upon checkpoint inhibition to prevent DNA replication stress. Our findings pave the way for patient selection in the clinical use of checkpoint inhibitors.


Prevention of influenza: recommendations for influenza immunization of children, 2007-2008.
The American Academy of Pediatrics recommends annual influenza immunization for all children with high-risk conditions who are 6 months of age and older, for all healthy children ages 6 through 59 months, for all household contacts and out-of-home caregivers of children with high-risk conditions and of healthy children younger than 5 years, and for all health care professionals. To more fully protect against the morbidity and mortality of influenza, increased efforts are needed to identify and immunize all children at high risk and all healthy children ages 6 through 59 months and to inform their parents when annual immunizations are due. Previously unimmunized children who are at least 6 months of age but younger than 9 years should receive 2 doses of influenza vaccine, given 1 month apart, beginning as soon as possible on the basis of local availability during the influenza season. If children in this cohort received only 1 dose for the first time in the previous season, it is recommended that 2 doses be administered in the current season. This recommendation applies only to the influenza season that follows the first year that a child younger than 9 years receives influenza vaccine. A child who then also fails to receive 2 doses the next year should be given only 1 dose per year from that point on. Influenza vaccine should also continue to be offered throughout the influenza season, even after influenza activity has been documented in a community. On the basis of global surveillance of circulating virus strains, the influenza vaccine may change from year to year; indeed, 1 of the 3 strains in the 2007-2008 vaccine is different from the previous year's vaccine. All health care professionals, influenza campaign organizers, and public health agencies should develop plans for expanding outreach and infrastructure to immunize all children for whom influenza vaccine is recommended. Appropriate prioritization of administering influenza vaccine will also be necessary when vaccine supplies are delayed or limited. Because the influenza season often extends into March, immunization against influenza is recommended to continue through late winter and early spring. Lastly, it is recommended that for the 2007-2008 season, and likely beyond, health care professionals do not prescribe amantadine or rimantadine for influenza treatment or chemoprophylaxis, because widespread resistance to these antiviral medications now exists among influenza A viral strains. However, oseltamivir and zanamivir can be prescribed for treatment or chemoprophylaxis, because influenza A and B strains remain susceptible.


Cadherin-11 expressed in association with mesenchymal morphogenesis in the head, somite, and limb bud of early mouse embryos.
Cadherin-11 (cad-11) is a novel member of the cadherin family of cell adhesion molecules, having recently been identified by means of the polymerase chain reaction. To study the function and expression of this molecule, we cloned mouse, cad-11 cDNA. Transfection of L cells with cDNA led them to acquire a typical cadherin-dependent cell-cell adhesiveness, and the L cells expressing cad-11 did not coaggregate with L cells expressing E-, P-, N-, or R-cadherin when they were mixed, indicating that this novel cadherin has a homophilic binding specificity, as found for other cadherins. To determine the developmental expression pattern of this molecule, we performed in situ hybridization analysis on early mouse embryos. Cad-11 first appeared in mesodermal layers only in the head and tail regions at the mid-to-late primitive streak stages. In the head, this appearance was followed by strong expression in mesenchymal tissues including branchial arches. In the trunk, the paraxial mesoderm initially did not express cad-11. However, as the somites formed, they expressed cad-11, and this expression was strictly correlated with their initial condensation and segregation from the presomitic mesoderm. The cad-11 expression in the somites was eventually restricted to sclerotome cells. As the limb buds developed, cad-11 appeared in the distal portion of the limb mesenchyme, and, at later stages, its expression was most evident at the peripheral mesenchyme. Cad-11 was thus expressed by restricted populations of mesenchymal cells in early embryos, although it was also expressed in parts of the neural tube, such as the optic vesicle and dorsal midline, and in part of the otic vesicle. As a step to investigate the role of cad-11 in mesenchymal cell adhesion, we dissociated the limb bud mesenchyme into single cells, pelleted them, and cultured them as aggregates. In these cultures, cad-11-positive cells clearly sorted out of the negative cell population, suggesting that cad-11 might be involved in selective association of mesenchymal cells. For comparison, we studied the expression of N-cadherin and found that the expressions of these two cadherins were differential, and complementary in some tissues. These results suggest that cad-11 is involved in specific associations of subsets of mesenchymal cells and also of some neural cells during early embryogenesis.


Stable and tunable phosphorescent neutral cyclometalated Au(III) diaryl complexes.
A series of novel luminescent cyclometalated Au(III) neutral complexes of the type cis-[(N(∧)C)AuL] [N(∧)C = 2-phenylpyridine (ppy), L = 1,1'-biphenyl (1)] and cis-[(N(∧)C)AuL(2)] [N(∧)C = 2-phenylpyridine (ppy), L = C(6)H(5) (2), C(6)F(5) (3), C(6)H(4)-CF(3)-p (4), 2-C(4)H(3)S (5)]; [N(∧)C = 2-(2-thienyl)pyridine (thpy), L = C(6)H(5) (6), C(6)F(5) (7)]; [N(∧)C = 2-(5-methyl-2-thienyl)pyridine (5 m-thpy), L = C(6)F(5) (8)] were successfully synthesized. The X-ray crystal structures of all compounds except 3 have been determined. These complexes were found to show long-lived emission in solution at room temperature. The emission origins of the complexes have been tentatively assigned to be derived from triplet states predominantly bearing intraligand (IL) character with some perturbation from the metal center. Density functional theory (DFT) calculations were performed to evaluate the stability associated with the complexes and TD-DFT calculations to ascertain the nature of the excited state. Variation of the cyclometalated ligands in the complexes readily leads to the tuning of the nature of the lower energy emissive states.


Discovery of dachshund 2 protein as a novel biomarker of poor prognosis in epithelial ovarian cancer.
The Dachshund homolog 2 (DACH2) gene has been implicated in development of the female genital tract in mouse models and premature ovarian failure syndrome, but to date, its expression in human normal and cancerous tissue remains unexplored. Using the Human Protein Atlas as a tool for cancer biomarker discovery, DACH2 protein was found to be differentially expressed in epithelial ovarian cancer (EOC). Here, the expression and prognostic significance of DACH2 was further evaluated in ovarian cancer cell lines and human EOC samples. Immunohistochemical expression of DACH2 was examined in tissue microarrays with 143 incident EOC cases from two prospective, population-based cohorts, including a subset of benign-appearing fallopian tubes (n = 32). A nuclear score (NS), i.e. multiplier of staining fraction and intensity, was calculated. For survival analyses, cases were dichotomized into low (NS < = 3) and high (NS > 3) using classification and regression tree analysis. Kaplan Meier analysis and Cox proportional hazards modelling were used to assess the impact of DACH2 expression on survival. DACH2 expression was analysed in the cisplatin sensitive ovarian cancer cell line A2780 and its cisplatin resistant derivative A2780-Cp70. The specificity of the DACH2 antibody was tested using siRNA-mediated silencing of DACH2 in A2780-Cp70 cells. DACH2 expression was considerably higher in the cisplatin resistant A2780-Cp70 cells compared to the cisplatin-sensitive A2780 cells. While present in all sampled fallopian tubes, DACH2 expression ranged from negative to strong in EOC. In EOC, DACH2 expression correlated with several proteins involved in DNA integrity and repair, and proliferation. DACH2 expression was significantly higher in carcinoma of the serous subtype compared to non-serous carcinoma. In the full cohort, high DACH2 expression was significantly associated with poor prognosis in univariable analysis, and in carcinoma of the serous subtype, DACH2 remained an independent factor of poor prognosis. This study provides a first demonstration of DACH2 protein being expressed in human fallopian tubes and EOC, with the highest expression in serous carcinoma where DACH2 was found to be an independent biomarker of poor prognosis. Future research should expand on the role of DACH2 in ovarian carcinogenesis and chemotherapy resistance.


[Duovac 300--udder health and milking routines. A field experiment (author's transl)].
The effect on udder health and milking routine of two Alfa Laval milking units, the Duovac 300 with a normal (380 mm Hg) and a low (250 mm Hg) vacuum phase and the standard HP 100 unit with only the normal (380 mm Hg) vacuum phase, was investigated on six dairy farms. A total of 197 cows were involved in the study. Each herd was divided into two groups which were as similar as possible in terms of age, stage of parturition and milk yield. One group was milked with the Duovac unit, while the other was milked with the HP 100 units for a test period of 100 days. No differences in cell counts from the cow samples, the presence of subclinical mastitis or clinical condition of the teats were observed between the two test groups. The total milking machine time for the Duovac units was longer than for the HP 100 units. However, the time spent in the normal vacuum phase (380 mm Hg) for the Duovac units was shorter than the total milking machine time for the HP 100 units (380 mm Hg). The Duovac units had the shortest machine stripping time, but the "waiting time" for the milkers (the time between the various operations) was longer. These time studies showed that the number of cows milked per hour by one man using four or five Duovac units was equal to or greater than the number of cows milked per hour by one man using three HP 100 units. It was recommended that no more than three HP 100 units be managed by one man.


Photoinduced interfacial charging and "explosion" of monolayer pentacene islands.
Electrostatic force microscopy shows that the electric field gradients above pentacene monolayer islands on 2-nm SiO2/Si substrates, in a dark, dry nitrogen environment, display a wide distribution of signs and magnitude that is dependent on sample history. Under 12 mW/cm2 green (532 nm) illumination, pentacene islands accumulate positive charge because of photoexcited electron transfer across the oxide to the Si substrate. At a strong illumination of 60 mW/cm2, pentacene islands reform into small spherical particles, apparently because the positive charge Coulomb repulsion energy becomes comparable to the cohesive energy of the pentacene monolayer.


Nanowire Aptasensors for Electrochemical Detection of Cell-Secreted Cytokines.
Cytokines are small proteins secreted by immune cells in response to pathogens/infections; therefore, these proteins can be used in diagnosing infectious diseases. For example, release of a cytokine interferon (IFN)-γ from T-cells is used for blood-based diagnosis of tuberculosis (TB). Our lab has previously developed an atpamer-based electrochemical biosensor for rapid and sensitive detection of IFN-γ. In this study, we explored the use of silicon nanowires (NWs) as a way to create nanostructured electrodes with enhanced sensitivity for IFN-γ. Si NWs were covered with gold and were further functionalized with thiolated aptamers specific for IFN-γ. Aptamer molecules were designed to form a hairpin and in addition to terminal thiol groups contained redox reporter molecules methylene blue. Binding of analyte to aptamer-modified NWs (termed here nanowire aptasensors) inhibited electron transfer from redox reporters to the electrode and caused electrochemical redox signal to decrease. In a series of experiments we demonstrate that NW aptasensors responded 3× faster and were 2× more sensitive to IFN-γ compared to standard flat electrodes. Most significantly, NW aptasensors allowed detection of IFN-γ from as few as 150 T-cells/mL while ELISA did not pick up signal from the same number of cells. One of the challenges faced by ELISA-based TB diagnostics is poor performance in patients whose T-cell numbers are low, typically HIV patients. Therefore, NW aptasensors developed here may be used in the future for more sensitive monitoring of IFN-γ responses in patients coinfected with HIV/TB.


Communication, cooperation, and social interactions: a report from the third Young Microbiologists Symposium on microbe signalling, organisation, and pathogenesis.
The third Young Microbiologists Symposium took place on the vibrant campus of the University of Dundee, Scotland, from the 2nd to 3rd of June 2014. The symposium attracted over 150 microbiologists from 17 different countries. The significant characteristic of this meeting was that it was specifically aimed at providing a forum for junior scientists to present their work. The meeting was supported by the Society for General Microbiology and the American Society for Microbiology, with further sponsorship from the European Molecular Biology Organization, the Federation of European Microbiological Societies, and The Royal Society of Edinburgh. In this report, we highlight some themes that emerged from the many exciting talks and poster presentations given by the young and talented microbiologists in the area of microbial gene expression, regulation, biogenesis, pathogenicity, and host interaction.


Benchmarking consensus model quality assessment for protein fold recognition.
Selecting the highest quality 3D model of a protein structure from a number of alternatives remains an important challenge in the field of structural bioinformatics. Many Model Quality Assessment Programs (MQAPs) have been developed which adopt various strategies in order to tackle this problem, ranging from the so called "true" MQAPs capable of producing a single energy score based on a single model, to methods which rely on structural comparisons of multiple models or additional information from meta-servers. However, it is clear that no current method can separate the highest accuracy models from the lowest consistently. In this paper, a number of the top performing MQAP methods are benchmarked in the context of the potential value that they add to protein fold recognition. Two novel methods are also described: ModSSEA, which based on the alignment of predicted secondary structure elements and ModFOLD which combines several true MQAP methods using an artificial neural network. The ModSSEA method is found to be an effective model quality assessment program for ranking multiple models from many servers, however further accuracy can be gained by using the consensus approach of ModFOLD. The ModFOLD method is shown to significantly outperform the true MQAPs tested and is competitive with methods which make use of clustering or additional information from multiple servers. Several of the true MQAPs are also shown to add value to most individual fold recognition servers by improving model selection, when applied as a post filter in order to re-rank models. MQAPs should be benchmarked appropriately for the practical context in which they are intended to be used. Clustering based methods are the top performing MQAPs where many models are available from many servers; however, they often do not add value to individual fold recognition servers when limited models are available. Conversely, the true MQAP methods tested can often be used as effective post filters for re-ranking few models from individual fold recognition servers and further improvements can be achieved using a consensus of these methods.


One-pot hydrothermal synthesis of mesoporous Zn(x)Cd(1-x)S/reduced graphene oxide hybrid material and its enhanced photocatalytic activity.
We successfully synthesized mesoporous Zn(x)Cd(1-x)S/reduced graphene oxide (Z(x)CSG) hybrid materials as photocatalysts using a facile one-pot hydrothermal reaction, in which graphene oxide (GO) was easily reduced (RGO), and simultaneously Zn(x)Cd(1-x)S (Z(x)CS) nanoparticles (NPs) with a mesoporous structure were uniformly dispersed on the RGO sheets. By well tuning the band gap from 3.42 to 2.21 eV by changing the molar ratio of Zn/Cd (or Zn content), Z(x)CSG with an optimal zinc content has been found to have a significant absorption in the visible light (VL) region. In addition, under VL irradiation (λ > 420 nm), Z(x)CSG also showed zinc content-dependent photocatalytic efficiencies for the degradation of methylene blue (MB). Our findings are that, among Z(x)CSG, Z(0.4)CSG displayed not only a superior photodegradation efficiency of MB (98%), but also good removal efficiency of total organic carbon (TOC) (67%). Furthermore, Z(0.4)CSG had a high photocatalytic stability, and could be used repeatedly. The enhanced photocatalytic activity for Z(0.4)CSG could be attributed to a synergistic effect between mesoporous Z(x)CS NPs and RGO, including the optimal band gap and the moderate conduction band position for ZxCS (compared to CdS), efficient separation and transfer ability of photogenerated electron/hole pairs in the presence of RGO sheets, and relatively high surface area for both mesoporous Z(x)CS NPs and RGO.


Living will completion in older adults.
In December 1991, the Patient Self-Determination Act required health care institutions to provide incoming patients with information about living wills (LWs). Although LWs have been legalized in the majority of states, the number of people with a completed LW remains low. This study was designed to learn some of the reasons that so few people have an LW. With the use of the health belief model, a questionnaire was designed to characterize the group that already had a completed LW and to identify the barriers older adults perceive in their completion of an LW. Older adults (aged 65 to 90 years) who dined at 10 local nutrition sites were asked to participate. Of the 214 subjects with usable responses, 15% already had executed an LW, 66% planned to complete an LW, and 86% wanted to have an LW. By means of multiple regression and discriminant analysis, the characteristics that defined each group were statistically determined. Two characteristics described those who already had an LW: they were highly educated and did not consider the LW form too long for its purpose. The older adults who planned to complete an LW identified two barriers impeding them: family issues and a need for assistance in completing the form. The majority of the older adults (61%) desired that their physicians initiate discussions with them about an LW. Conclusions from these data yield recommendations for health care providers toward implementing the act; physician-initiated discussions, community programs, and available information and assistance are needed.


ATAT1-enriched vesicles promote microtubule acetylation via axonal transport.
Microtubules are polymerized dimers of α- and β-tubulin that underlie a broad range of cellular activities. Acetylation of α-tubulin by the acetyltransferase ATAT1 modulates microtubule dynamics and functions in neurons. However, it remains unclear how this enzyme acetylates microtubules over long distances in axons. Here, we show that loss of ATAT1 impairs axonal transport in neurons in vivo, and cell-free motility assays confirm a requirement of α-tubulin acetylation for proper bidirectional vesicular transport. Moreover, we demonstrate that the main cellular pool of ATAT1 is transported at the cytosolic side of neuronal vesicles that are moving along axons. Together, our data suggest that axonal transport of ATAT1-enriched vesicles is the predominant driver of α-tubulin acetylation in axons.


Cardiac fibroblasts : Active players in (atrial) electrophysiology?
Fibrotic areas in cardiac muscle-be it in ventricular or atrial tissue-are considered as obstacles for conduction of the excitatory wave and can therefore facilitate re-entry, which may contribute to the sustenance of cardiac arrhythmias. Persistence of one of the most frequent arrhythmias, atrial fibrillation (AF), is accompanied by enhanced atrial fibrosis. Any kind of myocardial perturbation, whether via mechanical stress or ischemic damage, inflammation, or irregular and high-frequency electrical activity, activates fibroblasts. This leads to the secretion of paracrine factors and extracellular matrix proteins, especially collagen, and to the differentiation of fibroblasts into myofibroblasts. Excessive collagen production is the hallmark of fibrosis and impairs regular impulse propagation. In addition, direct electrical coupling between cardiomyocytes and nonmyocytes, such as fibroblasts and macrophages, via gap junctions affects conduction. Although fibroblasts are not electrically excitable, they express functional ion channels, in particular K+ channels and mechanosensitive channels, some of which could be involved in tissue remodeling. Here, we briefly review these aspects with special reference to AF.


Angiogenesis: an adaptive dynamic biological patterning problem.
Formation of functionally adequate vascular networks by angiogenesis presents a problem in biological patterning. Generated without predetermined spatial patterns, networks must develop hierarchical tree-like structures for efficient convective transport over large distances, combined with dense space-filling meshes for short diffusion distances to every point in the tissue. Moreover, networks must be capable of restructuring in response to changing functional demands without interruption of blood flow. Here, theoretical simulations based on experimental data are used to demonstrate that this patterning problem can be solved through over-abundant stochastic generation of vessels in response to a growth factor generated in hypoxic tissue regions, in parallel with refinement by structural adaptation and pruning. Essential biological mechanisms for generation of adequate and efficient vascular patterns are identified and impairments in vascular properties resulting from defects in these mechanisms are predicted. The results provide a framework for understanding vascular network formation in normal or pathological conditions and for predicting effects of therapies targeting angiogenesis.


Childhood tolerance of severe influenza: a mortality analysis in mice.
During the 1918 influenza pandemic, children experienced substantially lower mortality than adults, a striking but unexplained finding. Whether this was due to enhanced resistance (reduced virus load) or better tolerance (reduced impact of infection) has not been defined. We found that prepubertal mice infected with H1N1 influenza virus also showed greater survival than infected pubertal mice, despite similar virus loads. Transcriptome profiling of infected lungs identified estrogen as a regulator of susceptibility in both sexes and also linked better survival to late expression of IL-1β. Blocking puberty with gonadectomy or a gonadotropin-releasing hormone antagonist improved survival. Estrogen or testosterone (which can be converted to estrogen) restored susceptibility of gonadectomized pubertal mice to influenza mortality, but dihydrotestosterone (which cannot be converted to estrogen) did not. Estrogen receptor blockade with fulvestrant in both male and female pubertal mice resulted in improved survival, even when given 3 days after infection. Moreover, late, but not early, IL-1β neutralization after infection was also protective. These findings indicate that pubertal increases in estrogen in both sexes are associated with increased mortality during influenza. This helps explain the reduced mortality of children seen with influenza in 1918 and might also be relevant to childhood tolerance to many other infectious diseases.


Structure of Escherichia coli K-12 miaA and characterization of the mutator phenotype caused by miaA insertion mutations.
Previously, we reported several unusual relationships between the 2-methylthio-N6-(delta 2-isopentenyl)adenosine-37 (ms2i6A-37) tRNA modification and spontaneous mutagenesis in Escherichia coli K-12 (D. M. Connolly and M. E. Winkler, J. Bacteriol. 171:3233-3246, 1989). To confirm and extend these observations, we determined the structure of miaA, which mediates the first step of ms2i6A-37 synthesis, and characterized the miaA mutator phenotype. The most likely translation start of miaA overlaps the last two codons of mutL, which encodes a protein required for methyl-directed mismatch repair. This structural arrangement confirms that miaA and mutL are in the same complex operon. The miaA gene product, delta 2-isopentenylpyrophosphate transferase, shows extensive homology with the yeast MOD5 gene product, and both enzymes contain a substrate binding site found in farnysyl pyrophosphate synthetase and a conserved putative ATP/GTP binding site. Insertions in miaA cause exclusively GC----TA transversions, which contrasts with the GC----AT and AT----GC transitions observed in mutL mutants. To correlate the absence of the ms2i6A-37 tRNA modification directly with the mutator phenotype, we isolated a unique suppressor of a leaky miaA(ochre) mutation. The miaD suppressor mapped to 99.75 min, restored the ms2i6A-37 tRNA modification to miaA(ochre) mutants, and abolished the miaA mutator phenotype. We speculate that miaD causes a decrease in ms2i6A-37 tRNA demodification or an increase in miaA gene expression but not at the level of operon transcription. Together, these observations support the idea that the ms2i6A-37 tRNA modification acts as a physiological switch that modulates spontaneous mutation frequency and other metabolic functions.


The German version of the Assessment of Chronic Illness Care: instrument translation and cultural adaptation.
In Switzerland, there is a dearth of information on the extent to which patients with chronic illnesses receive care congruent with the Chronic Care Model (CCM). To drive quality improvement programmes, it is necessary to have practical assessment tools in the country's own language to evaluate the delivery of CCM activities. German translation and adaptation of the original Assessment of Chronic Illness Care (ACIC). We followed a sequential forward and backward translation approach. In a multidisciplinary committee review the original English version and the translations were compared, instructions and formats modified and cross-cultural equivalences verified. The second version was pre-tested and multidisciplinary group discussion led to the final version which aimed to create a comprehensive culturally adapted translation capturing the original idea of the items rather than a direct one to one translation. Difficulties encountered during the translation process consisted in the difference of health care settings and health care organization in Switzerland and USA.The adapted German version was delivered to a managed care organization in the city of Zurich to test the initial use for diabetes care. The average ACIC subscale scores were: organization of the health care delivery system: mean (m) = 7.31 (SD = 0.79), community linkages: m = 3.78 (SD = 1.09), self-management support: m = 4.88 (SD = 1.21), decision support: m = 4.79 (SD = 1.16), delivery system design: m = 5.56 (SD = 1.28) and clinical information systems: m = 4.50 (SD = 2.69). Overall, the ACIC subscale scores were comparable with the scores of the original testing. After cultural adaptations the German version of the ACIC is applicable as a tool to guide quality improvement in chronic illness care in German speaking countries in Europe.


Triple therapy of hepatocellular carcinoma with microRNA-122 and doxorubicin co-loaded functionalized gold nanocages.
A combination of different therapy strategies has great potential to efficaciously treat malignant tumors, by virtue of their synergetic effects. Herein, a co-delivery system based on gold nanocages (AuNCs) was designed to deliver both doxorubicin (DOX) and microRNA-122 mimic (miR-122) for an enhanced cancer therapy. DOX was loaded into the AuNCs and miR-122 was condensed onto the surface of the functionalized AuNCs by an electrostatic interaction. Polyethyleneglycol (PEG) and hyaluronic acid (HA) were also introduced to the co-delivery system for targeted drug delivery. We evaluated the cellular uptake, biodistribution and anti-tumor effect in vitro and in vivo. Our results demonstrated an effective delivery of DOX and miR-122 into tumor cells and the tumor tissue. Importantly, the triple therapy, namely the combination of chemotherapy, gene therapy and photothermal therapy, mediated by this multifunctional drug delivery system, exhibited better anti-tumor effect than any single therapy, both in vitro and in vivo. Additionally, this drug delivery system caused insignificant toxicity to the major organs and had no obvious effect on the body weight of the mice. It could be concluded that multifunctional AuNCs are promising as a co-delivery vector for an enhanced anti-tumor effect.


Assessment of environmental and surgical mask contamination at a student health center - 2012-2013 influenza season.
Increased understanding of influenza transmission is critical for pandemic planning and selecting appropriate controls for healthcare personnel safety and health. The goals of this pilot study were to assess environmental contamination in different areas and at two time periods in the influenza season and to determine the feasibility of using surgical mask contamination to evaluate potential exposure to influenza virus. Bioaerosol samples were collected over 12 days (two 6-day sessions) at 12 locations within a student health center using portable two-stage bioaerosol samplers operating 8 hr each day. Surface samples were collected each morning and afternoon from common high-contact non-porous hard surfaces from rooms and locations where bioaerosol samplers were located. Surgical masks worn by participants while in contact with patients with influenza-like illness were collected. A questionnaire administered to each of the 12 participants at the end of each workday and another at the end of each workweek assessed influenza-like illness symptoms, estimated the number of influenza-like illness patient contacts, hand hygiene, and surgical mask usage. All samples were analyzed using qPCR. Over the 12 days of the study, three of the 127 (2.4%) bioaerosol samples, 2 of 483 (0.41%) surface samples, and 0 of 54 surgical masks were positive for influenza virus. For the duration of contact that occurred with an influenza patient on any of the 12 days, nurse practitioners and physicians reported contacts with influenza-like illness patients >60 min, medical assistants reported 15-44 min, and administrative staff reported <30 min. Given the limited number of bioaerosol and surface samples positive for influenza virus in the bioaerosol and surface samples, the absence of influenza virus on the surgical masks provides inconclusive evidence for the potential to use surgical masks to assess exposure to influenza viruses. Further studies are needed to determine feasibility of this approach in assessing healthcare personnel exposures. Information learned in this study can inform future field studies on influenza transmission.


Proteomic responses to gold(iii)-toxicity in the bacterium Cupriavidus metallidurans CH34.
The metal-resistant β-proteobacterium Cupriavidus metallidurans drives gold (Au) biomineralisation and the (trans)formation of Au nuggets largely via unknown biochemical processes, ultimately leading to the reductive precipitation of mobile, toxic Au(i/iii)-complexes. In this study proteomic responses of C. metallidurans CH34 to mobile, toxic Au(iii)-chloride are investigated. Cells were grown in the presence of 10 and 50 μM Au(iii)-chloride, 50 μM Cu(ii)-chloride and without additional metals. Differentially expressed proteins were detected by difference gel electrophoresis and identified by liquid chromatography coupled mass spectrometry. Proteins that were more abundant in the presence of Au(iii)-chloride are involved in a range of important cellular functions, e.g., metabolic activities, transcriptional regulation, efflux and metal transport. To identify Au-binding proteins, protein extracts were separated by native 2D gel electrophoresis and Au in protein spots was detected by laser absorption inductively coupled plasma mass spectrometry. A chaperon protein commonly understood to bind copper (Cu), CupC, was identified and shown to bind Au. This indicates that it forms part of a multi-metal detoxification system and suggests that similar/shared detoxification pathways for Au and Cu exist. Overall, this means that C. metallidurans CH34 is able to mollify the toxic effects of cytoplasmic Au(iii) by sequestering this Au-species. This effect may in the future be used to develop CupC-based biosensing capabilities for the in-field detection of Au in exploration samples.


Condensing results of wet sieving analyses into a single data: a comparison of methods for particle size description.
Sieve analysis is used in feed analysis, and studies of digestive physiology with various approaches to describe an average value of particle size which can serve to compare different samples. To demonstrate the effects of such different approaches, we compared five particle size indicators to demonstrate advantages and disadvantages of each method, the modulus of fineness (MOF), the discrete mean (dMEAN) and median (dMED), and the continuous mean (cMEAN) and median (cMED), well aware of the fact that a gold standard for this procedure is lacking. Data were obtained from 580 individual faecal samples of different herbivore species by wet sieving over a cascade of nine sieves with mesh sizes ranging from 0.063 to 16 mm. MOF, dMEAN and dMED can be calculated directly from the results of sieve analysis, but cMEAN and cMED require a curve-fitting procedure. Across the whole sample size, dMEAN and cMEAN showed the highest correlation. The correlation between the respective MEAN and MED was higher for d than for c. As expected, MOF deviated most from the other measurements. Simulating different sieve sets resulted in a poor correlation between the results from the different sets in MOF and cMED, but a good correlation in dMEAN and cMEAN, suggesting that these latter measures can also be compared between studies that do not use identical sieve sets. As the calculation of dMEAN is comparatively simpler and less time-consuming than that of cMEAN, we propose the dMEAN as a standard for the description of a mean particle size value obtained from sieve analysis. For practical application, the good correlation of different simulated sieve sets indicates that sets with fewer sieves could be used in large-scale studies to reduce analytical workload.


Stabilization of atherosclerotic plaques by blockade of macrophage migration inhibitory factor after vascular injury in apolipoprotein E-deficient mice.
Macrophage migration inhibitory factor (MIF), a cytokine that controls cell-mediated inflammatory responses, is upregulated in atherogenesis; however, its functional contribution to lesion development has not been evaluated. We studied the role of MIF on neointima lesion formation after wire-induced injury of carotid arteries in apolipoprotein E-deficient (apoE(-/-)) mice. Immunohistochemistry revealed that MIF expression was detectable in endothelial cells before injury and upregulated in smooth muscle cells (SMCs) 24 hours after endothelial denudation. Three weeks after injury, MIF was predominantly found in endothelial cells and macrophage-derived foam cells. Neutralizing MIF with a monoclonal antibody resulted in a marked reduction of neointimal macrophages and inhibited transformation of macrophages into foam cells. Conversely, the content of SMCs and of collagen in the neointima were increased, amounting to a slight but not significant reduction in neointima and media size after 3 weeks of MIF monoclonal antibody treatment. Notably, serum levels of the cytokines IL-2, IL-4, IL-6, IL-10, and tumor necrosis factor were increased in MIF monoclonal antibody-treated mice. In vitro flow assays revealed that MIF pretreatment of aortic endothelium enhanced monocyte recruitment and that the monocyte arrest induced by oxidized LDL is mediated by endothelial MIF, as shown by monoclonal antibody inhibition. Inhibition of MIF resulted in a shift in the cellular composition of neointimal plaques toward a stabilized phenotype with reduced macrophage/foam cell content and increased SMC content. This might be attributable to a reduction of monocyte recruitment mediated by endothelial MIF.


Pain Duration and Resolution following Surgery: An Inception Cohort Study.
Preoperative determinants of pain duration following surgery are poorly understood. We identified preoperative predictors of prolonged pain after surgery in a mixed surgical cohort. We conducted a prospective longitudinal study of patients undergoing mastectomy, lumpectomy, thoracotomy, total knee replacement, or total hip replacement. We measured preoperative psychological distress and substance use, and then measured pain and opioid use after surgery until patients reported the cessation of both opioid consumption and pain. The primary endpoint was time to opioid cessation, and those results have been previously reported. Here, we report preoperative determinants of time to pain resolution following surgery in Cox proportional hazards regression. Between January 2007 and April 2009, we enrolled 107 of 134 consecutively approached patients undergoing the aforementioned surgical procedures. In the final multivariate model, preoperative self-perceived risk of addiction predicted more prolonged pain. Unexpectedly, anxiety sensitivity predicted more rapid pain resolution after surgery. Each one-point increase (on a four point scale) of self-perceived risk of addiction was associated with a 38% (95% CI 3-61) reduction in the rate of pain resolution (P = 0.04). Furthermore, higher anxiety sensitivity was associated with an 89% (95% CI 23-190) increased rate of pain resolution (P = 0.004). Greater preoperative self-perceived risk of addiction, and lower anxiety sensitivity predicted a slower rate of pain resolution following surgery. Each of these factors was a better predictor of pain duration than preoperative depressive symptoms, post-traumatic stress disorder symptoms, past substance use, fear of pain, gender, age, preoperative pain, or preoperative opioid use.


Population genomics of invasive rodents on islands: Genetic consequences of colonization and prospects for localized synthetic gene drive.
Introduced rodent populations pose significant threats worldwide, with particularly severe impacts on islands. Advancements in genome editing have motivated interest in synthetic gene drives that could potentially provide efficient and localized suppression of invasive rodent populations. Application of such technologies will require rigorous population genomic surveys to evaluate population connectivity, taxonomic identification, and to inform design of gene drive localization mechanisms. One proposed approach leverages the predicted shifts in genetic variation that accompany island colonization, wherein founder effects, genetic drift, and island-specific selection are expected to result in locally fixed alleles (LFA) that are variable in neighboring nontarget populations. Engineering of guide RNAs that target LFA may thus yield gene drives that spread within invasive island populations, but would have limited impacts on nontarget populations in the event of an escape. Here we used pooled whole-genome sequencing of invasive mouse (Mus musculus) populations on four islands along with paired putative source populations to test genetic predictions of island colonization and characterize locally fixed Cas9 genomic targets. Patterns of variation across the genome reflected marked reductions in allelic diversity in island populations and moderate to high degrees of differentiation from nearby source populations despite relatively recent colonization. Locally fixed Cas9 sites in female fertility genes were observed in all island populations, including a small number with multiplexing potential. In practice, rigorous sampling of presumptive LFA will be essential to fully assess risk of resistance alleles. These results should serve to guide development of improved, spatially limited gene drive design in future applications.


Role of fluoride varnish in preventing early childhood caries: A systematic review.
Early childhood caries is a public health problem that continues to affect babies and preschool children worldwide. This untreated caries process results in progressive destruction of the crowns of the teeth, often accompanied by severe pain and suffering, affecting the quality of life. Fluoride varnish which is one of the most important materials to prevent ECC is easy to apply and well tolerated by children. This study aimed to evaluate the scientific evidence regardingthe role of fluoride varnish in preventing early childhood caries. Records were searched from various databases such as PubMed/Medline, Cochrane, and EMBASE. Articles published over the past 36 years (1979-2015) were identified using the key search terms. A total of 190 records were identified by title/abstracts/full text articles and were retrieved. Potentially relevant reports identified from the reference lists of relevant studies, review articles and chapters were hand-searched, which yielded an additional 10 articles. The main outcome of our investigation was prevention of early childhood caries following application of fluoride varnish and unavoidable fluoride exposure. Out of 190 articles originally identified, 30 records were considered potentially eligible and sought for further assessment. 17 articles met the inclusion criteria and these studies were assessed independently for methodology and performance. Analysis of literature revealed that basically two concentrations of fluoride varnishes have been used: 1% and 5%, with a caries preventive fraction ranges of 6.4-30% and 5-63%, respectively. The results showed that fluoride varnishes have been used at concentrations of 1% and 5% in the prevention of ECC. The preventive fraction was influenced by the frequency of application, the duration of study and sample size. The evidence level of the studies was of moderate to limited value.


Can Temporal Fine Structure and Temporal Envelope be Considered Independently for Pitch Perception?
In psychoacoustics, works on pitch perception attempt to distinguish between envelope and fine structure cues that are generally viewed as independent and separated using a Hilbert transform. To empirically distinguish between envelope and fine structure cues in pitch perception experiments, a dedicated signal has been proposed. This signal is an unresolved harmonic complex tones with all harmonics shifted by the same amount of Hz. As the frequency distance between adjacent components is regular and identical than in the original harmonic complex tone, such a signal has the same envelope but a different fine structure. So, any perceptual difference between these signals is interpreted as a fine structure based percept. Here, as illustrated by very basic simulations, I suggest that this orthogonal point of view that is generally admitted could be a conceptual error. In fact, neither the fine structure nor the envelope is required to be fully encoded to explain pitch perception. Sufficient information is conveyed by the peaks in the fine structure that are located nearby a maximum of the envelope. Envelope and fine structure could then be in perpetual interaction and the pitch would be conveyed by "the fine structure under envelope". Moreover, as the temporal delay between peaks of interest is rather longer than the delay between two adjacent peaks of the fine structure, such a mechanism would be much less constrained by the phase locking limitation of the auditory system. Several data from the literature are discussed from this new conceptual point of view.


Outcome analysis for women undergoing annual versus biennial screening mammography: a review of 24,211 examinations.
Our goal was to determine differences in outcome measures between women undergoing annual versus biennial screening mammography. A retrospective review of prospectively collected data on 24,211 consecutive screening mammography examinations was performed in women aged 40-79 years, all of whom had undergone previous normal screening mammography. Annual screening and biennial screening were defined as examinations performed 10-14 months and 22-26 months, respectively, after previous normal screening mammography. The rates of recall, biopsy, cancer detection, and interval cancer for annual and biennial screening cohorts were calculated, as were tumor size, lymph node status, and stage of invasive cancer. Interval cancer cases were identified by linkage with a regional tumor registry. Of the 4306 biennially screened women, 160 were recalled (3.7%), 45 were biopsied (1.0%), and cancer was detected in 19 (0.44%). Of the 19,905 annually screened women, 518 were recalled (2.6%), 150 were biopsied (0.75%), and cancer was detected in 71 (0.36%). Of the 3278 registry-linked biennially screened women, five had interval cancer (0.15%); of the 15,031 registry-linked annually screened women, 10 had interval cancer (0.07%). For biennial screening-detected cancer and interval invasive cancer combined, the median tumor size was 15 mm, 24% had lymph node metastasis, and 29% were stage 2 or higher. For annual screening-detected cancer, these measures were 11 mm, 14% positive nodes, and 17% stage 2+ cancer, respectively. Annual screening mammography results in lower recall rates than does biennial screening (p < .0001). Moreover, annual screening results in the detection of smaller tumors that have a more favorable prognosis (p = .04).


Molecular pathology shows p16 methylation in nonadenomatous pituitaries from patients with Cushing's disease.
The majority of cases of Cushing's disease are due to the presence of a corticotroph microadenoma. Less frequently no adenoma is found and histology shows either corticotroph hyperplasia, or apparently normal pituitary. In this study we have used molecular pathology to determine whether the tissue labeled histologically as "normal" is indeed abnormal. Tissue from 31 corticotroph adenomas and 16 nonadenomatous pituitaries were subject to methylation-sensitive PCR to determine the methylation status of the p16 gene CpG island. The proportion of methylated versus unmethylated CpG island was determined using combined bisulphite restriction analysis. Methylation status was correlated with immunohistochemical detection of p16. Seventeen of 31 adenomas (54.8%), 4 of 6 cases of corticotroph hyperplasia, and 7 of 10 apparently normal pituitaries showed p16 methylation. Ten of 14 (71%; P = 0.01) adenomas and 2 of 3 cases of corticotroph hyperplasia, which were methylated, failed to express p16 protein. However, only 2 of 7 apparently normal pituitaries that were methylated failed to express p16 protein. Quantitative analysis of methylation using combined bisulphite restriction analysis showed only unmethylated CpG islands in postmortem normal pituitaries; however, in adenomas 80-90% of the cells within a specimen were methylated. The reverse was true for corticotroph hyperplasia and apparently normal pituitaries where only 10-20% of the cells were methylated. Thus, the decreased proportion of cells that were methylated, particularly in those cases of apparently normal pituitary, is the most likely explanation for the lack of association between this change and loss of cognate protein in these cases. To our knowledge this is the first report that describes an intrinsic molecular change, namely methylation of the p16 gene CpG island, common to all three histological patterns associated with Cushing's disease. Thus, the use of molecular pathology reveals abnormalities undetected by routine pathological investigation. In cases of "apparently" normal pituitaries it is not possible to determine whether the change is associated with adenoma cells "scattered" throughout the gland, albeit few in number, or with the ancestor-clonal origin of these tumor cells.


Characterization of the megakaryocyte demarcation membrane system and its role in thrombopoiesis.
To produce blood platelets, megakaryocytes elaborate proplatelets, accompanied by expansion of membrane surface area and dramatic cytoskeletal rearrangements. The invaginated demarcation membrane system (DMS), a hallmark of mature cells, has been proposed as the source of proplatelet membranes. By direct visualization of labeled DMS, we demonstrate that this is indeed the case. Late in megakaryocyte ontogeny, the DMS gets loaded with PI-4,5-P(2), a phospholipid that is confined to plasma membranes in other cells. Appearance of PI-4,5-P(2) in the DMS occurs in proximity to PI-5-P-4-kinase alpha (PIP4Kalpha), and short hairpin (sh) RNA-mediated loss of PIP4Kalpha impairs both DMS development and expansion of megakaryocyte size. Thus, PI-4,5-P(2) is a marker and possibly essential component of internal membranes. PI-4,5-P(2) is known to promote actin polymerization by activating Rho-like GTPases and Wiskott-Aldrich syndrome (WASp) family proteins. Indeed, PI-4,5-P(2) in the megakaryocyte DMS associates with filamentous actin. Expression of a dominant-negative N-WASp fragment or pharmacologic inhibition of actin polymerization causes similar arrests in proplatelet formation, acting at a step beyond expansion of the DMS and cell mass. These observations collectively suggest a signaling pathway wherein PI-4,5-P(2) might facilitate DMS development and local assembly of actin fibers in preparation for platelet biogenesis.


Boosting facility deliveries with results-based financing: a mixed-methods evaluation of the government midwifery incentive scheme in Cambodia.
Increasing the coverage of skilled attendance at births in a health facility (facility delivery) is crucial for saving the lives of mothers and achieving Millennium Development Goal five. Cambodia has significantly increased the coverage of facility deliveries and reduced the maternal mortality ratio in the last decade. The introduction of a nationwide government implemented and funded results-based financing initiative, known as the Government Midwifery Incentive Scheme (GMIS), is considered one of the most important contributors to this. We evaluated GMIS to explore its effects on facility deliveries and the health system. We used a mixed-methods design. An interrupted time series model was applied, using routine longitudinal data on reported deliveries between 2006 and 2011 that were extracted from the health information system. In addition, we interviewed 56 key informants and performed 12 focus group discussions with 124 women who had given birth (once or more) since 2006. Findings from the quantitative data were carefully interpreted and triangulated with those from qualitative data. We found that facility deliveries have tripled from 19% of the estimated number of births in 2006 to 57% in 2011 and this increase was more substantial at health centres as compared to hospitals. Segmented linear regressions showed that the introduction of GMIS in October 2007 made the increase in facility deliveries and deliveries with skilled attendants significantly jump by 18 and 10% respectively. Results from qualitative data also suggest that the introduction of GMIS together with other interventions that aimed to improve access to essential maternal health services led to considerable improvements in public health facilities and a steep increase in facility deliveries. Home deliveries attended by traditional birth attendants decreased concomitantly. We also outline several operational issues and limitations of GMIS. The available evidence strongly suggests that GMIS is an effective mechanism to complement other interventions to improve health system performance and boost facility deliveries as well as skilled birth attendance; thereby contributing to the reduction of maternal mortality. Our findings provide useful lessons for Cambodia to further improve GMIS and for other low-income countries to implement similar results-based financing mechanisms.


Satellite-based decadal change assessments of pan-Arctic environments.
Remote sensing can advance the work of the Circumpolar Biodiversity Monitoring Program through monitoring of satellite-derived terrestrial and marine physical and ecological variables. Standardized data facilitate an unbiased comparison across variables and environments. Using MODIS standard products of land surface temperature, percent snow covered area, NDVI, EVI, phenology, burned area, marine chlorophyll, CDOM, sea surface temperature, and marine primary productivity, significant trends were observed in almost all variables between 2000 and 2017. Analysis of seasonal data revealed significant breakpoints in temporal trends. Within the terrestrial environment, data showed significant increasing trends in land surface temperature and NDVI. In the marine environment, significant increasing trends were detected in primary productivity. Significantly earlier onset of green up date was observed in bioclimate subzones C&E and longer end of growing season in B&E. Terrestrial and marine parameters showed similar rates of change with unidirectional change in terrestrial and significant directional and magnitude shifts in marine.


High sodium diet converts renal proteoglycans into pro-inflammatory mediators in rats.
High dietary sodium aggravates renal disease by affecting blood pressure and by its recently shown pro-inflammatory and pro-fibrotic effects. Moreover, pro-inflammatory modification of renal heparan sulfate (HS) can induce tissue remodeling. We aim to investigate if high sodium intake in normotensive rats converts renal HS into a pro-inflammatory phenotype, able to bind more sodium and orchestrate inflammation, fibrosis and lymphangiogenesis. Wistar rats received a normal diet for 4 weeks, or 8% NaCl diet for 2 or 4 weeks. Blood pressure was monitored, and plasma, urine and tissue collected. Tissue sodium was measured by flame spectroscopy. Renal HS and tubulo-interstitial remodeling were studied by biochemical, immunohistochemical and qRT-PCR approaches. High sodium rats showed a transient increase in blood pressure (week 1; p<0.01) and increased sodium excretion (p<0.05) at 2 and 4 weeks compared to controls. Tubulo-interstitial T-cells, myofibroblasts and mRNA levels of VCAM1, TGF-β1 and collagen type III significantly increased after 4 weeks (all p<0.05). There was a trend for increased macrophage infiltration and lymphangiogenesis (both p = 0.07). Despite increased dermal sodium over time (p<0.05), renal concentrations remained stable. Renal HS of high sodium rats showed increased sulfation (p = 0.05), increased L-selectin binding to HS (p<0,05), and a reduction of sulfation-sensitive anti-HS mAbs JM403 (p<0.001) and 10E4 (p<0.01). Hyaluronan expression increased under high salt conditions (p<0.01) without significant changes in the chondroitin sulfate proteoglycan versican. Statistical analyses showed that sodium-induced tissue remodeling responses partly correlated with observed HS changes. We show that high salt intake by healthy normotensive rats convert renal HS into high sulfated pro-inflammatory glycans involved in tissue remodeling events, but not in increased sodium storage.


The evolutionary convergence of hearing in a parasitoid fly and its cricket host.
Parasitism is a widespread and diverse life strategy that connects species throughout the animal kingdom. Female parasitoid flies of the genus Ormia must find a specific cricket host on which to deposit their parasitic maggots. To reproduce, female flies must perform the same task as female crickets: find a singing male cricket. These flies have evolved a unique hearing organ that allows them to detect and locate singing male crickets. Through evolutionary convergence, these flies possess a hearing organ that much more resembles a cricket's ear than a typical fly's ear, allowing these parasitoids to take advantage of the sensory ecological niche of their host.


Antioxidants for male subfertility.
Between 30% to 80% of male subfertility cases are considered to be due to the damaging effects of oxidative stress on sperm and 1 man in 20 will be affected by subfertility. Antioxidants are widely available and inexpensive when compared to other fertility treatments and many men are already using these to improve their fertility. It is thought that oral supplementation with antioxidants may improve sperm quality by reducing oxidative stress. Pentoxifylline, a drug that acts like an antioxidant, was also included in this review. This Cochrane review aimed to evaluate the effectiveness and safety of oral supplementation with antioxidants for subfertile male partners in couples seeking fertility assistance. We searched the Cochrane Menstrual Disorders and Subfertility Group Specialised Register, CENTRAL, MEDLINE, EMBASE, CINAHL, PsycINFO and AMED databases (from inception until January 2014); trial registers; sources of unpublished literature and reference lists. An updated search was run in August 2014 when potentially eligible studies were placed in 'Studies awaiting assessment'. We included randomised controlled trials (RCTs) comparing any type or dose of antioxidant supplement (single or combined) taken by the subfertile male partner of a couple seeking fertility assistance with a placebo, no treatment or another antioxidant. Two review authors independently selected eligible studies, extracted the data and assessed the risk of bias of the included studies. The primary review outcome was live birth; secondary outcomes included clinical pregnancy rates, adverse events, sperm DNA fragmentation, sperm motility and concentration. Data were combined, where appropriate, to calculate pooled odds ratios (ORs) or mean differences (MD) and 95% confidence intervals (CIs). Statistical heterogeneity was assessed using the I(2) statistic. We assessed the overall quality of the evidence for the main outcomes using GRADE methods. This updated review included 48 RCTs that compared single and combined antioxidants with placebo, no treatment or another antioxidant in a population of 4179 subfertile men. The duration of the trials ranged from 3 to 26 weeks with follow up ranging from 3 weeks to 2 years. The men were aged from 20 to 52 years. Most of the men enrolled in these trials had low total sperm motility and sperm concentration. One study enrolled men after varicocelectomy, one enrolled men with a varicocoele, and one recruited men with chronic prostatitis. Three trials enrolled men who, as a couple, were undergoing in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) and one trial enrolled men who were part of a couple undergoing intrauterine insemination (IUI). Funding sources were stated by 15 trials. Four of these trials stated that funding was from a commercial source and the remaining 11 obtained funding through non-commercial avenues or university grants. Thirty-three trials did not report any funding sources.A limitation of this review was that in a sense we had included two different groups of trials, those that reported on the use of antioxidants and the effect on live birth and clinical pregnancy, and a second group that reported on sperm parameters as their primary outcome and had no intention of reporting the primary outcomes of this review. We included 25 trials reporting on sperm parameters and only three of these reported on live birth or clinical pregnancy. Other limitations included poor reporting of study methods, imprecision, the small number of trials providing usable data, the small sample size of many of the included studies and the lack of adverse events reporting. The evidence was graded as 'very low' to 'low'. The data were current to 31 January 2014.Live birth: antioxidants may have increased live birth rates (OR 4.21, 95% CI 2.08 to 8.51, P< 0.0001, 4 RCTs, 277 men, I(2) = 0%, low quality evidence). This suggests that if the chance of a live birth following placebo or no treatment is assumed to be 5%, the chance following the use of antioxidants is estimated to be between 10% and 31%. However, this result was based on only 44 live births from a total of 277 couples in four small studies.Clinical pregnancy rate: antioxidants may have increased clinical pregnancy rates (OR 3.43, 95% CI 1.92 to 6.11, P < 0.0001, 7 RCTs, 522 men, I(2) = 0%, low quality evidence). This suggests that if the chance of clinical pregnancy following placebo or no treatment is assumed to be 6%, the chance following the use of antioxidants is estimated at between 11% and 28%. However, there were only seven small studies in this analysis and the quality of the evidence was rated as low.Miscarriage: only three trials reported on this outcome and the event rate was very low. There was insufficient evidence to show whether there was a difference in miscarriage rates between the antioxidant and placebo or no treatment groups (OR 1.74, 95% CI 0.40 to 7.60, P = 0.46, 3 RCTs, 247 men, I(2) = 0%, very low quality evidence). The findings suggest that in a population of subfertile men with an expected miscarriage rate of 2%, use of an antioxidant would result in the risk of a miscarriage lying between 1% and 13%.Gastrointestinal upsets: there was insufficient evidence to show whether there was a difference in gastrointestinal upsets when antioxidants were compared to placebo or no treatment as the event rate was very low (OR 1.60, 95% CI 0.47 to 5.50, P = 0.46, 6 RCTs, 429 men, I(2) = 0%).We were unable to draw any conclusions from the antioxidant versus antioxidant comparison as not enough trials compared the same interventions. There is low quality evidence from only four small randomised controlled trials suggesting that antioxidant supplementation in subfertile males may improve live birth rates for couples attending fertility clinics. Low quality evidence suggests that clinical pregnancy rates may increase. There is no evidence of increased risk of miscarriage but this is uncertain as the evidence is of very low quality. Data were lacking on other adverse effects. Further large well-designed randomised placebo-controlled trials are needed to clarify these results.


Group II intron protein localization and insertion sites are affected by polyphosphate.
Mobile group II introns consist of a catalytic intron RNA and an intron-encoded protein with reverse transcriptase activity, which act together in a ribonucleoprotein particle to promote DNA integration during intron mobility. Previously, we found that the Lactococcus lactis Ll.LtrB intron-encoded protein (LtrA) expressed alone or with the intron RNA to form ribonucleoprotein particles localizes to bacterial cellular poles, potentially accounting for the intron's preferential insertion in the oriC and ter regions of the Escherichia coli chromosome. Here, by using cell microarrays and automated fluorescence microscopy to screen a transposon-insertion library, we identified five E. coli genes (gppA, uhpT, wcaK, ynbC, and zntR) whose disruption results in both an increased proportion of cells with more diffuse LtrA localization and a more uniform genomic distribution of Ll.LtrB-insertion sites. Surprisingly, we find that a common factor affecting LtrA localization in these and other disruptants is the accumulation of intracellular polyphosphate, which appears to bind LtrA and other basic proteins and delocalize them away from the poles. Our findings show that the intracellular localization of a group II intron-encoded protein is a major determinant of insertion-site preference. More generally, our results suggest that polyphosphate accumulation may provide a means of localizing proteins to different sites of action during cellular stress or entry into stationary phase, with potentially wide physiological consequences.


The neural basis of sex differences in sexual behavior: A quantitative meta-analysis.
Sexuality as to its etymology presupposes the duality of sexes. Using quantitative neuroimaging meta-analyses, we demonstrate robust sex differences in the neural processing of sexual stimuli in thalamus, hypothalamus, and basal ganglia. In a narrative review, we show how these relate to the well-established sex differences on the behavioral level. More specifically, we describe the neural bases of known poor agreement between self-reported and genital measures of female sexual arousal, of previously proposed male proneness to affective sexual conditioning, as well as hints of unconscious activation of bonding mechanisms during sexual stimulation in women. In summary, our meta-analytic review demonstrates that neurofunctional sex differences during sexual stimulation can account for well-established sex differences in sexual behavior.


A SSR-based composite genetic linkage map for the cultivated peanut (Arachis hypogaea L.) genome.
The construction of genetic linkage maps for cultivated peanut (Arachis hypogaea L.) has and continues to be an important research goal to facilitate quantitative trait locus (QTL) analysis and gene tagging for use in a marker-assisted selection in breeding. Even though a few maps have been developed, they were constructed using diploid or interspecific tetraploid populations. The most recently published intra-specific map was constructed from the cross of cultivated peanuts, in which only 135 simple sequence repeat (SSR) markers were sparsely populated in 22 linkage groups. The more detailed linkage map with sufficient markers is necessary to be feasible for QTL identification and marker-assisted selection. The objective of this study was to construct a genetic linkage map of cultivated peanut using simple sequence repeat (SSR) markers derived primarily from peanut genomic sequences, expressed sequence tags (ESTs), and by "data mining" sequences released in GenBank. Three recombinant inbred lines (RILs) populations were constructed from three crosses with one common female parental line Yueyou 13, a high yielding Spanish market type. The four parents were screened with 1044 primer pairs designed to amplify SSRs and 901 primer pairs produced clear PCR products. Of the 901 primer pairs, 146, 124 and 64 primer pairs (markers) were polymorphic in these populations, respectively, and used in genotyping these RIL populations. Individual linkage maps were constructed from each of the three populations and a composite map based on 93 common loci were created using JoinMap. The composite linkage maps consist of 22 composite linkage groups (LG) with 175 SSR markers (including 47 SSRs on the published AA genome maps), representing the 20 chromosomes of A. hypogaea. The total composite map length is 885.4 cM, with an average marker density of 5.8 cM. Segregation distortion in the 3 populations was 23.0%, 13.5% and 7.8% of the markers, respectively. These distorted loci tended to cluster on LG1, LG3, LG4 and LG5. There were only 15 EST-SSR markers mapped due to low polymorphism. By comparison, there were potential synteny, collinear order of some markers and conservation of collinear linkage groups among the maps and with the AA genome but not fully conservative. A composite linkage map was constructed from three individual mapping populations with 175 SSR markers in 22 composite linkage groups. This composite genetic linkage map is among the first "true" tetraploid peanut maps produced. This map also consists of 47 SSRs that have been used in the published AA genome maps, and could be used in comparative mapping studies. The primers described in this study are PCR-based markers, which are easy to share for genetic mapping in peanuts. All 1044 primer pairs are provided as additional files and the three RIL populations will be made available to public upon request for quantitative trait loci (QTL) analysis and linkage map improvement.


A profile of anti-vaccination lobbying on the South African internet, 2011-2013.
The South African Vaccination and Immunisation Centre receives many requests to explain the validity of internet-based anti-vaccination claims. Previous global studies on internet-based anti-vaccination lobbying had not identified anti-vaccination web pages originating in South Africa (SA). To characterise SA internet-based anti-vaccination lobbying. In 2011, searches for anti-vaccination content were performed using Google, Yahoo and MSN-Bing, limited to English-language SA web pages. Content analysis was performed on web pages expressing anti-vaccination sentiment about infant vaccination. This was repeated in 2012 and 2013 using Google, with the first 700 web pages per search being analysed. Blogs/forums, articles and e-shops constituted 40.3%, 55.2% and 4.5% of web pages, respectively. Authors were lay people (63.5%), complementary/alternative medicine (CAM) practitioners (23.1%), medical professionals practising CAM (7.7%) and medical professionals practising only allopathic medicine (5.8%). Advertisements appeared on 55.2% of web pages. Of these, 67.6% were sponsored by or linked to organisations with financial interests in discrediting vaccines, with 80.0% and 24.0% of web pages sponsored by these organisations claiming respectively that vaccines are ineffective and that vaccination is profit driven. The vast majority of web pages (92.5%) claimed that vaccines are not safe, and 77.6% of anti-vaccination claims originated from the USA. South Africans are creating web pages or blogs for local anti-vaccination lobbying. Research is needed to understand what influence internet-based anti-vaccination lobbying has on the uptake of infant vaccination in SA.


Ischemia-induced stimulation of Na-K-Cl cotransport in cerebral microvascular endothelial cells involves AMP kinase.
Increased blood-brain barrier (BBB) Na-K-Cl cotransporter activity appears to contribute to cerebral edema formation during ischemic stroke. We have shown previously that inhibition of BBB Na-K-Cl cotransporter activity reduces edema and infarct in the rat middle cerebral artery occlusion (MCAO) model of ischemic stroke. We have also shown that the BBB cotransporter is stimulated by the ischemic factors hypoxia, aglycemia, and arginine vasopressin (AVP), although the mechanisms responsible are not well understood. AMP-activated protein kinase (AMPK), a key mediator of cell responses to stress, can be activated by a variety of stresses, including ischemia, hypoxia, and aglycemia. Previous studies have shown that the AMPK inhibitor Compound C significantly reduces infarct in mouse MCAO. The present study was conducted to evaluate the possibility that AMPK participates in ischemic factor-induced stimulation of the BBB Na-K-Cl cotransporter. Cerebral microvascular endothelial cells (CMEC) were assessed for Na-K-Cl cotransporter activity as bumetanide-sensitive (86)Rb influx. AMPK activity was assessed by Western blot analysis and immunofluorescence methods using antibodies that detect total versus phosphorylated (activated) AMPK. We found that hypoxia (7% and 2% O(2)), aglycemia, AVP, and oxygen-glucose deprivation (5- to 120-min exposures) increase activation of AMPK. We also found that Compound C inhibition of AMPK reduces hypoxia-, aglycemia-, and AVP-induced stimulation of CMEC Na-K-Cl cotransporter activity. Confocal immunofluorescence of perfusion-fixed rat brain slices revealed the presence of AMPK, both total and phosphorylated kinase, in BBB in situ of both control and ischemic brain. These findings suggest that ischemic factor stimulation of the BBB Na-K-Cl cotransporter involves activation of AMPK.


Comparative epigenomic analysis of murine and human adipogenesis.
We report the generation and comparative analysis of genome-wide chromatin state maps, PPARγ and CTCF localization maps, and gene expression profiles from murine and human models of adipogenesis. The data provide high-resolution views of chromatin remodeling during cellular differentiation and allow identification of thousands of putative preadipocyte- and adipocyte-specific cis-regulatory elements based on dynamic chromatin signatures. We find that the specific locations of most such elements differ between the two models, including at orthologous loci with similar expression patterns. Based on sequence analysis and reporter assays, we show that these differences are determined, in part, by evolutionary turnover of transcription factor motifs in the genome sequences and that this turnover may be facilitated by the presence of multiple distal regulatory elements at adipogenesis-dependent loci. We also utilize the close relationship between open chromatin marks and transcription factor motifs to identify and validate PLZF and SRF as regulators of adipogenesis.


Dispersive liquid-liquid microextraction: trends in the analysis of biological samples.
Dispersive liquid-liquid microextraction (DLLME) is a recent microextraction technique that was first developed by Rezaee and co-workers in 2006. It allows the simultaneous extraction and preconcentration of analytes into a micro-volume of extracting solvent based on a ternary solvent system involving an aqueous phase, a nonpolar water immiscible high-density solvent that acts as extraction phase, and a disperser solvent, which is often polar and water miscible. This article presents an overview of DLLME applications in the analysis of biological samples (e.g., plasma and urine). Aside from the classical DLLME applications using high density extraction solvents, recent advances in the use of low density solvents and ionic liquids are also discussed. Although most of the applications deal with the analysis of organic target compounds, a few applications on the bioanalysis of inorganic substances are also included.


Rectal adenocarcinoma with choriocarcinomatous differentiation: clinical and genetic aspects.
Nongestational choriocarcinomas are rare tumors. In the gastrointestinal tract, they are characterized by a biphasic tumor growth with separated areas of adenocarcinomatous and choriocarcinomatous differentiation. We here report a case of a combined adenocarcinoma-choriocarcinoma of the rectum. The tumor showed an aggressive clinical behavior with metastasis to the liver and lungs. A transient partial remission was achieved after 4 cycles of cisplatinum, etoposide, and ifosfamide chemotherapy, with normalization of serum beta-human chorionic gonadotropin levels. At this time, viable residual choriocarcinoma cells were found in surgically resected lung metastasis. The patient succumbed 8 months after initial diagnosis to a rapid abdominal relapse. We used comparative genomic hybridization (CGH) and fluorescence in situ hybridization to elucidate the genetic relationship of adenocarcinoma and choriocarcinoma in this neoplasm. We found genetic changes characteristic for colorectal adenocarcinomas, a loss of chromosomal regions 8p21-pter as well as 18q21-pter, and a gain of 5p and 20q, in both tumor parts. This provides evidence for the common origin of both components. A differential pattern of additional genetic changes suggests a clonal evolution from a common ancestor cell. In contrast to findings from a comparative study on a choriocarcinoma of the renal pelvis, we did not find an amplification of the germ cell cancer-associated chromosomal region 12p11.2-p12.1 in the areas of choriocarcinoma but found instead a loss of Xp11.3-pter. To our knowledge, this is the first report of a CGH comparison of the adenocarcinomatous and choriocarcinomatous tumor parts in a nongestational choriocarcinoma of the gastrointestinal tract.


Oral health inequalities among indigenous and nonindigenous children in the Northern Territory of Australia.
To describe oral health inequalities among indigenous and nonindigenous children in the Northern Territory of Australia using an area-based measure of socioeconomic status (SES). Data were obtained from indigenous and nonindigenous 4-13-year-old children enrolled in the Northern Territory School Dental Service in 2002-2003. The Socio-Economic Indices For Areas (SEIFA) were used to determine socioeconomic relationships with dental disease experience. Some 12,584 children were examined, 35.1% of whom were indigenous. Across all age-groups, socially disadvantaged indigenous children experienced higher mean dmft and DMFT levels than their similarly aged, similarly disadvantaged nonindigenous counterparts. Indigenous children aged 5 years had almost four times the dmft of their nonindigenous counterparts in the same disadvantage category (P < 0.05), while indigenous children aged 10 years had almost five times the DMFT of similarly disadvantaged nonindigenous children (P < 0.05). A distinct social gradient was apparent among indigenous and nonindigenous children, respectively, whereby those with the highest dmft/DMFT levels were in the most disadvantaged SES category and those least disadvantaged had the lowest dmft/DMFT levels. In most age-groups, indigenous children who were least disadvantaged had worse oral health than the most disadvantaged nonindigenous children. The findings suggest that indigenous status and SES have strong oral health outcome correlations but are not mutually dependent, that is, indigenous status influences oral health outcomes irrespective of social disadvantage. From a health policy perspective, greater oral health gains may be possible by concentrating public health and clinical effort among all indigenous children irrespective of SES status.


Are you Scared Yet?: Evaluating Fear Appeal Messages in Tweets about the Tips Campaign.
In March 2012, the CDC launched "Tips from Former Smokers," a $54 million national campaign featuring individuals experiencing long-term health consequences of smoking. The campaign approach was based on strong evidence that anti-tobacco ads portraying fear, graphic images, and personal testimonials are associated with attitudinal and behavior change. Yet it was also controversial; critics cited the danger that viewers might reject such intensely graphic messages. Tasked with informing this debate, our study analyzes the corpus of Tips campaign-related tweets obtained via the Twitter Firehose. We provide a novel and rigorous method for media campaign evaluation within the framework of the Extended Parallel Process Model. Among the relevant Tweets, 87% showed evidence of message acceptance, while 7% exhibited message rejection.