Second-trimester routine ultrasound screening: expectations and experiences in a nationwide Swedish sample.
To investigate, in a large nationwide Swedish sample, pregnant women's expectations of the routine second-trimester ultrasound examination, with participants expressing themselves in their own words, and to determine whether they had been given sufficient information about why and how the examination was performed, and about possible risks. We focused specifically on reasons for women not having a positive experience. Of 4600 eligible Swedish-speaking women, 3061 were recruited to the study in early pregnancy, during three 1-week periods spread evenly over 1 year (1999-2000), and these women completed a questionnaire at a mean of 16 weeks' gestation. A follow-up questionnaire at 2 months after delivery was completed by 2730 women. The representativeness of the sample was assessed by comparison with the total Swedish birth cohort of 1999. The most prominent expectation about the up-coming scan was confirmation that the baby was well, followed by confirmation that the pregnancy was real. Detailed information, such as date of delivery and sex of the baby, was mentioned less often, and very few wrote about the examination as an exciting and joyful experience. After the birth, a large majority was satisfied with information about why (88%) and how (87%) the examination was performed, but only 58% said they had received sufficient information about possible risks. 94% had a positive experience of the scan, and those who had not had more ambivalent feelings about their pregnancy. Women with negative feelings about the scan were more often single and of non-Swedish background, and emotional problems were more common in this group. Women's expectations of the routine second-trimester scan differ from those of caregivers, focusing on general reassurance rather than specific information. Level of satisfaction with the scan was high, but information given about risks could be improved. Women with ambivalent or negative feelings about pregnancy may have difficulties enjoying the examination.


Negative refraction in Photonic Crystals: thickness dependence and Pendellösung phenomenon.
We show that the refracted wave at the exit surface of a Photonic Crystal (PhC) slab is periodically modulated, in positive or in negative direction, changing the slab thickness. In spite of an always increasing literature, the effect of the thickness in negative refraction on PhC's does not seem to be appropriately considered. However such an effect is not surprising if interpreted with the help of Dynamical Diffraction Theory (DDT), which is generally applied in the x-ray diffraction. The thickness dependence is a direct result of the so-called Pendellösung phenomenon. That explains the periodic exchange, inside the crystal, of the energy among direct beam (or positively refracted) and diffracted beam (or negatively refracted). The Pendellösung phenomenon is an outstanding example of the application of the DDT as a powerful and simple tool for the analysis of s electromagnetic interaction in PhC's.


Structure-activity studies of tylosin-related macrolides.
The effects of tylosin-related macrolide antibiotics were examined in cell-free protein synthesis (using a coupled transcription-translation system derived from Streptomyces lividans) and against whole cells of that organism. Anti-ribosomal potency was determined primarily by the number and nature of the glycosyl substituents, and was not significantly influenced by lactone ring oxidation or sugar methylation. In contrast, uptake of the drugs into S. lividans was influenced, either positively or negatively, by each of these structural parameters. The presence of erm type I or erm type II resistance genes in S. lividans markedly affected the resistance phenotype and studies involving ribosomes from such strains revealed differences in macrolide activity that were not otherwise apparent.


High biocompatibility and improved osteogenic potential of amorphous calcium carbonate/vaterite.
In human bone, amorphous calcium carbonate (ACC) is formed as a precursor of the crystalline carbonated apatite/hydroxyapatite (HA). Here we describe that the metastable ACC phase can be stabilized by inorganic polyphosphate (polyP) that is also used as a phosphate source for the non-enzymatic carbonate/phosphate exchange during HA formation. This polymer was found to suppress the transformation of ACC into crystalline CaCO3 at a percentage of 5% [w/w] ("CCP5") with respect to CaCO3 and almost completely at 10% [w/w] ("CCP10"). Both preparations (CaCO3/polyP) are amorphous, but also contain small amounts of vaterite, as revealed by XRD, FTIR and SEM analyses. They did not affect the growth/viability of SaOS-2 cells. Cell culture and Ca2+ release experiments revealed that the CaCO3 particles formed in the presence of polyP (CaCO3/polyP) are degradable and, unlike calcite, become disintegrated with time during the cell culture incubation. Again in contrast to calcite, "CCP5" and "CCP10" were found to exhibit osteogenic activity and induce the expression of alkaline phosphatase gene in SaOS-2 cells as well as in human mesenchymal stem cells (MSC). In vivo studies in rats, using PLGA microspheres inserted in the muscles of the back of the animals, revealed that the encapsulated "CCP10" is not only biocompatible but also supports the regeneration at the implant region. We conclude that ACC containing small amounts of vaterite has osteogenic potential and offers superior properties compared to the biologically inert calcite with respect to a potential application as a scaffold material for bone implants.


Role of mechanical power estimates in the O2 cost of walking in children with cerebral palsy.
It has been established in able-bodied children that traditional biomechanical descriptors of gait such as stride length or stride frequency do not fully account for the differences seen in the energy cost of locomotion noted with age. Hence, measures of total body mechanical power output have been adopted to explain these differences. The aim of this study was to estimate the ability of this mechanical power calculation to explain the variability in the metabolic energy cost of treadmill walking in children with spastic cerebral palsy (CP). Thirteen subjects volunteered for the study. One group consisted of eight (6 male, 2 female) children with CP (age 12.2 +/- 2.7 yr). The second group consisted of five (4 male, 1 female) able-bodied controls (age 13.4 +/- 2.8 yr). The treadmill walking protocol consisted of one 4-min stage at 0% grade, 3 km x h(-1). Infrared markers were placed on 12 anatomical landmarks and data were collected using the OPTOTRAK motion analysis system over a 5-s time period during the last 30 s of the 4-min stage. On-line oxygen consumption VO2 measurements were obtained throughout using the Beckman Horizon Metabolic Cart. Relative VO2 (mL x kg(-1) x min(-1)) was significantly (P < 0.05) different between the two groups (CP: 16.6 +/- 6.5 vs control: 10.2 +/- 1.2). Simple linear regression analysis demonstrated that mechanical power measurements, incorporating transfers of energy between and within adjacent body segments, accounted for 87.2% of the total variability noted in VO2 for the children with CP, compared with only 2.4% in the able-bodied subjects. The results indicate that mechanical power differences explain the majority of the variability noted in VO2 in children with CP at a submaximal walking speed.


High-resolution structures of Lactobacillus salivarius transketolase in the presence and absence of thiamine pyrophosphate.
Probiotic bacterial strains have been shown to enhance the health of the host through a range of mechanisms including colonization, resistance against pathogens, secretion of antimicrobial compounds and modulation of the activity of the innate immune system. Lactobacillus salivarius UCC118 is a well characterized probiotic strain which survives intestinal transit and has many desirable host-interaction properties. Probiotic bacteria display a wide range of catabolic activities, which determine their competitiveness in vivo. Some lactobacilli are heterofermentative and can metabolize pentoses, using a pathway in which transketolase and transaldolase are key enzymes. L. salivarius UCC118 is capable of pentose utilization because it encodes the key enzymes on a megaplasmid. The crystal structures of the megaplasmid-encoded transketolase with and without the enzyme cofactor thiamine pyrophosphate have been determined. Comparisons with other known transketolase structures reveal a high degree of structural conservation in both the catalytic site and the overall conformation. This work extends structural knowledge of the transketolases to the industrially and commercially important Lactobacillus genus.


Interictal PET and ictal subtraction SPECT: sensitivity in the detection of seizure foci in patients with medically intractable epilepsy.
Interictal positron emission tomography (PET) and ictal subtraction single photon emission computed tomography (SPECT) of the brain have been shown to be valuable tests in the presurgical evaluation of epilepsy. To determine the relative utility of these methods in the localization of seizure foci, we compared interictal PET and ictal subtraction SPECT to subdural and depth electrode recordings in patients with medically intractable epilepsy. Between 2003 and 2009, clinical information on all patients at our institution undergoing intracranial electroencephalography (EEG) monitoring was charted in a prospectively recorded database. Patients who underwent preoperative interictal PET and ictal subtraction SPECT were selected from this database. Patient characteristics and the findings on preoperative interictal PET and ictal subtraction SPECT were analyzed. Sensitivity of detection of seizure foci for each modality, as compared to intracranial EEG monitoring, was calculated. Fifty-three patients underwent intracranial EEG monitoring with preoperative interictal PET and ictal subtraction SPECT scans. The average patient age was 32.7 years (median 32 years, range 1-60 years). Twenty-seven patients had findings of reduced metabolism on interictal PET scan, whereas all 53 patients studied demonstrated a region of relative hyperperfusion on ictal subtraction SPECT suggestive of an epileptogenic zone. Intracranial EEG monitoring identified a single seizure focus in 45 patients, with 39 eventually undergoing resective surgery. Of the 45 patients in whom a seizure focus was localized, PET scan identified the same region in 25 cases (56% sensitivity) and SPECT in 39 cases (87% sensitivity). Intracranial EEG was concordant with at least one study in 41 cases (91%) and both studies in 23 cases (51%). In 16 (80%) of 20 cases where PET did not correlate with intracranial EEG, the SPECT study was concordant. Conversely, PET and intracranial EEG were concordant in two (33%) of the six cases where the SPECT did not demonstrate the seizure focus outlined by intracranial EEG. Thirty-three patients had surgical resection and >2 years of follow-up, and 21 of these (64%) had Engel class 1 outcome. No significant effect of imaging concordance on seizure outcome was seen. Interictal PET and ictal subtraction SPECT studies can provide important information in the preoperative evaluation of medically intractable epilepsy. Of the two studies, ictal subtraction SPECT appears to be the more sensitive. When both studies are used together, however, they can provide complementary information.


Recent advances in understanding the genetics of sleep.
Sleep is a ubiquitous and complex behavior in both its manifestation and regulation. Despite its essential role in maintaining optimal performance, health, and well-being, the genetic mechanisms underlying sleep remain poorly understood. Here, we review the forward genetic approaches undertaken in the last four years to elucidate the genes and gene pathways affecting sleep and its regulation. Despite an increasing number of studies and mining large databases, a coherent picture on "sleep" genes has yet to emerge. We highlight the results achieved by using unbiased genetic screens mainly in humans, mice, and fruit flies with an emphasis on normal sleep and make reference to lessons learned from the circadian field.


Seasonal copepod lipid pump promotes carbon sequestration in the deep North Atlantic.
Estimates of carbon flux to the deep oceans are essential for our understanding of global carbon budgets. Sinking of detrital material ("biological pump") is usually thought to be the main biological component of this flux. Here, we identify an additional biological mechanism, the seasonal "lipid pump," which is highly efficient at sequestering carbon into the deep ocean. It involves the vertical transport and metabolism of carbon rich lipids by overwintering zooplankton. We show that one species, the copepod Calanus finmarchicus overwintering in the North Atlantic, sequesters an amount of carbon equivalent to the sinking flux of detrital material. The efficiency of the lipid pump derives from a near-complete decoupling between nutrient and carbon cycling—a "lipid shunt," and its direct transport of carbon through the mesopelagic zone to below the permanent thermocline with very little attenuation. Inclusion of the lipid pump almost doubles the previous estimates of deep-ocean carbon sequestration by biological processes in the North Atlantic.


Material characterization of the brainstem from oscillatory shear tests.
Traumatic damage to the brainstem occurs frequently when the brain skull complex experiences injurious loading especially during those traumatic situations that produce diffuse axonal injury (DAI). DAI has been shown to be dependent on load direction and correlated with regional tissue deformation in response to rotational inertial loads. Possible mechanisms for the selective vulnerability of the brainstem are (1) the geometry of the central nervous system is responsible for producing high tissue strains in these regions, (2) regional differences in overall material stiffness result in larger deformations at these sites, and (3) the anisotropic mechanical properties of these regions lead to a sensitivity to the rotational load direction and magnitude. This paper investigates the latter two hypotheses by performing oscillatory shear tests on adult porcine brainstem in three mutually perpendicular directions. The complex shear moduli were calculated over a range of frequencies (20-200 Hz), for three levels of peak engineering strain (2.5%, 5.0%, and 7.5%). The directional data demonstrated that the brainstem exhibits significant transversely isotropic behavior. Both components of the complex modulus in which the axonal fibers are oriented parallel to the plane of shear but transverse to the shear direction were significantly higher than those of the other two, mutually indistinguishable test cases across the range of strains tested. By comparison with similar tests on cerebral tissue, these data demonstrated that the brainstem displays a stiffer biomechanical response. These differences were present for both components of the complex shear modulus and were greater as the magnitude of the applied strain increased. The regional stiffness and anisotropic response of the brainstem coupled with its location as a narrow bridge between CNS regions interact to result in the selective vulnerability of this region in rotational loading.


Conformation of receptor-bound visual arrestin.
Arrestin-1 (visual arrestin) binds to light-activated phosphorylated rhodopsin (P-Rh*) to terminate G-protein signaling. To map conformational changes upon binding to the receptor, pairs of spin labels were introduced in arrestin-1 and double electron-electron resonance was used to monitor interspin distance changes upon P-Rh* binding. The results indicate that the relative position of the N and C domains remains largely unchanged, contrary to expectations of a "clam-shell" model. A loop implicated in P-Rh* binding that connects β-strands V and VI (the "finger loop," residues 67-79) moves toward the expected location of P-Rh* in the complex, but does not assume a fully extended conformation. A striking and unexpected movement of a loop containing residue 139 away from the adjacent finger loop is observed, which appears to facilitate P-Rh* binding. This change is accompanied by smaller movements of distal loops containing residues 157 and 344 at the tips of the N and C domains, which correspond to "plastic" regions of arrestin-1 that have distinct conformations in monomers of the crystal tetramer. Remarkably, the loops containing residues 139, 157, and 344 appear to have high flexibility in both free arrestin-1 and the P-Rh*complex.


Functional analysis of plastid DNA replication origins in tobacco by targeted inactivation.
Sequences described as chloroplast DNA replication origins were analysed in vivo by creating deletion and insertion mutants via plastid transformation in tobacco. Deletion of the described oriA sequence, which is located within the intron of the trnI gene, resulted in heteroplastomic transformants, when the selection marker was inserted within the intron. Removal of the complete intron sequence together with the oriA sequence, however, yielded homoplastomic transformants of normal phenotype, in which wild-type signals were no longer detectable through Southern analysis, thus bringing the role of the described oriA sequence for plastome replication into question. Similarly, deletion of sequence elements upstream of trnI, which have a possible ori function in Oenothera, did not show any effect in tobacco. The two copies of oriB, which are located at the very end of the plastome Inverted Repeats, were targeted with two different transformation vectors in a cotransformation approach. While in initial transformants integration of the selection marker could be detected at both sites, the transgene was found exclusively at one site or the other after additional rounds of regeneration. Whereas the copy of oriB in Inverted Repeat B could be completely deleted, targeting of the copy in Inverted Repeat A resulted in heteroplastomic lines, as the essential ycf1 gene was also affected. Due to the strong selection against cotransformants we conclude that at least one copy of the oriB sequence is essential for plastome replication, whereas replication appears possible without oriA elements.


The possible "proton sponge " effect of polyethylenimine (PEI) does not include change in lysosomal pH.
Polycations such as polyethylenimine (PEI) are used in many novel nonviral vector designs and there are continuous efforts to increase our mechanistic understanding of their interactions with cells. Even so, the mechanism of polyplex escape from the endosomal/lysosomal pathway after internalization is still elusive. The "proton sponge " hypothesis remains the most generally accepted mechanism, although it is heavily debated. This hypothesis is associated with the large buffering capacity of PEI and other polycations, which has been interpreted to cause an increase in lysosomal pH even though no conclusive proof has been provided. In the present study, we have used a nanoparticle pH sensor that was developed for pH measurements in the endosomal/lysosomal pathway. We have carried out quantitative measurements of lysosomal pH as a function of PEI content and correlate the results to the "proton sponge " hypothesis. Our measurements show that PEI does not induce change in lysosomal pH as previously suggested and quantification of PEI concentrations in lysosomes makes it uncertain that the "proton sponge " effect is the dominant mechanism of polyplex escape.


Anomalous sulphur isotopes in plume lavas reveal deep mantle storage of Archaean crust.
Basaltic lavas erupted at some oceanic intraplate hotspot volcanoes are thought to sample ancient subducted crustal materials. However, the residence time of these subducted materials in the mantle is uncertain and model-dependent, and compelling evidence for their return to the surface in regions of mantle upwelling beneath hotspots is lacking. Here we report anomalous sulphur isotope signatures indicating mass-independent fractionation (MIF) in olivine-hosted sulphides from 20-million-year-old ocean island basalts from Mangaia, Cook Islands (Polynesia), which have been suggested to sample recycled oceanic crust. Terrestrial MIF sulphur isotope signatures (in which the amount of fractionation does not scale in proportion with the difference in the masses of the isotopes) were generated exclusively through atmospheric photochemical reactions until about 2.45 billion years ago. Therefore, the discovery of MIF sulphur in these young plume lavas suggests that sulphur--probably derived from hydrothermally altered oceanic crust--was subducted into the mantle before 2.45 billion years ago and recycled into the mantle source of Mangaia lavas. These new data provide evidence for ancient materials, with negative Δ(33)S values, in the mantle source for Mangaia lavas. Our data also complement evidence for recycling of the sulphur content of ancient sedimentary materials to the subcontinental lithospheric mantle that has been identified in diamond-hosted sulphide inclusions. This Archaean age for recycled oceanic crust also provides key constraints on the length of time that subducted crustal material can survive in the mantle, and on the timescales of mantle convection from subduction to upwelling beneath hotspots.


Fluorine impurities at CeO2(111): Effects on oxygen vacancy formation, molecular adsorption, and surface re-oxidation.
We investigate the effects of anion doping with fluorine impurities on the chemistry of the CeO2 (111) facet, using the results of DFT + U calculations. We consider three prototypical processes: the formation of oxygen vacancies, the adsorption of O2 and H2O molecules, and the re-oxidation of the surface with fragments of the two molecules. We find that the first two of these processes are not strongly affected, but that the presence of F lowers the energy gained in the re-oxidation of the surface in comparison to the healing of an oxygen vacancy, by 1.47 eV in the case of O2 (provided that the F is part of a cluster) and by 0.92 eV in the case of H2O. Based on these results, we suggest that F could enhance the redox chemistry of ceria by toggling between being in the surface and on the surface, effectively facilitating the release of lattice O by acting as a "place holder" for it. Finally, we find that the desorption of F as either 12F2 or HF is energetically unfavourable, suggesting that F doped ceria should be stable in the presence of O2 and H2O.


Noninvasive characterization of continuous-profile blazed diffraction gratings.
Smooth, continuous-profile diffractive optical elements and other regular microstructures demand advanced characterization methods. Here the problem of reconstructing the profiles with optical scatterometry is addressed for the first time to our knowledge. We represent the profile of a blazed grating with an analytic Chebyshev series. The coefficients of the series are inferred from the far-field diffraction pattern, when the grating profile is illuminated with coherent light. An advanced neural-network model is used as a nonlinear statistical estimator. The reconstruction error is found to be less than 4% as normalized to the depth of the profile, even with realistic measurement errors included in the simulations.


Strategically addressing the nurse shortage: a closer look at the nurse funders collaborative.
Despite consistent public and private investments in nursing over several decades, nurse shortages persist, appearing more acute today than ever before. The Nurse Funders Collaborative, a group of foundations, government agencies, and corporations convened by the Robert Wood Johnson Foundation, has been meeting since 2003, seeking opportunities to address issues facing nursing and health care more strategically. This paper reports on a study conducted under the collaborative's auspices, which highlights the categorical and regional funding patterns of funders of nursing over five years. This information provides nursing funders with ways to craft new solutions to the nurse shortage.


Who teaches science to nurses?
To explore the demographics of individuals teaching basic science courses in private nursing schools. The study was conducted in Karachi, and comprised teachers teaching basic science courses in 16 registered private nursing schools. A demographic questionnaire was used to collect data. The study was conducted in the year 2013. Teachers holding academic/professional degrees in different science disciplines were involved in teaching science to nurses. In most of the schools, all the basic science courses were taught by one subject specialist science teacher. These subject specialist science teachers held degrees in different disciplines, which included Doctor of Philosophy, Master of Philosophy, Masters in Science, Bachelor of Medicine, Bachelor of Surgery, Bachelors in Engineering, Doctor of Pharmacy and registered nurse with a master's degree. Except the physicist(2), eleven (11) subject specialists taught two or more than two science courses. Microbiologist (4) and physiologist (5) were generally engaged in teaching all science subjects. Science courses in nursing can be taught by anyone holding a bachelor or a master's degree in science or a professional degree.


Increasing BCI communication rates with dynamic stopping towards more practical use: an ALS study.
The P300 speller is a brain-computer interface (BCI) that can possibly restore communication abilities to individuals with severe neuromuscular disabilities, such as amyotrophic lateral sclerosis (ALS), by exploiting elicited brain signals in electroencephalography (EEG) data. However, accurate spelling with BCIs is slow due to the need to average data over multiple trials to increase the signal-to-noise ratio (SNR) of the elicited brain signals. Probabilistic approaches to dynamically control data collection have shown improved performance in non-disabled populations; however, validation of these approaches in a target BCI user population has not occurred. We have developed a data-driven algorithm for the P300 speller based on Bayesian inference that improves spelling time by adaptively selecting the number of trials based on the acute SNR of a user's EEG data. We further enhanced the algorithm by incorporating information about the user's language. In this current study, we test and validate the algorithms online in a target BCI user population, by comparing the performance of the dynamic stopping (DS) (or early stopping) algorithms against the current state-of-the-art method, static data collection, where the amount of data collected is fixed prior to online operation. Results from online testing of the DS algorithms in participants with ALS demonstrate a significant increase in communication rate as measured in bits/min (100-300%), and theoretical bit rate (100-550%), while maintaining selection accuracy. Participants also overwhelmingly preferred the DS algorithms. We have developed a viable BCI algorithm that has been tested in a target BCI population which has the potential for translation to improve BCI speller performance towards more practical use for communication.


Interactive effects of pressurized ventilation, water depth and substrate conditions on Phragmites australis.
Pressurized ventilation acts to increase the oxygen supply to roots and rhizomes in some species of emergent plants. In a greenhouse experiment, we investigated how pressurized ventilation affected growth, biomass allocation and mineral content of Phragmites australis in two water depths (15 cm or 75 cm) and two substrates (organic sediment or sand). Through perforating each stem above the water surface, pressurized ventilation was inhibited without affecting oxygen diffusion. In controls, 10-20% of the stems were perforated to make certain that lack of efflux sites would not limit pressurized ventilation. Plants with inhibited pressurized ventilation had lower oxygen concentrations in their stem bases than control plants. Growth was lower in plants with inhibited pressurized ventilation compared to controls except when plants grew in a combination of sand and shallow water. In plants grown in an organic sediment, but not in those grown in sand, inhibition of pressurized ventilation resulted in decreased biomass allocation to soil roots but increased allocation to aquatic roots. Stem perforation affected the tissue concentrations of nitrogen, phosphorus, magnesium, manganese and aluminium but not of calcium or iron. We suggest that the lower growth in plants with inhibited pressurized ventilation was caused by decreased mineral uptake, which may have resulted from the decreased proportional allocation to soil root weight, from decreased mineral availability or from impaired root function. In plants grown in sand in shallow water, diffusion seemed to cover the oxygen demand, as pressurized ventilation did not affect growth.


A survey of the prevalence of diabetes type 2 amongst schizophrenics in a chronic care treatment facility.
The determination of the prevalence of type 2 diabetes in a group of patients suffering from schizophrenia who have been hospitalized in a chronic care facility and to consider the role of factors which may contribute to the increased risk of developing diabetes. The hospital files of patients suffering from diabetes type 2 were reviewed and assessments were made in terms of age, gender and body mass index (BMI) duration of stay and co-existing medical conditions. Fasting blood glucose estimations were done on all schizophrenic patients not receiving treatment for diabetes. Medications of patients were recorded. A protocol was prepared and approval to conduct the study was obtained from the hospital authorities. Of 494 schizophrenic patients 19 (3.85%) were found to suffer from diabetes. 68% of patients in the diabetic group were found to be overweight in terms of BMI rating. Only one patient had a medical condition. Proportionately the number of women was relatively high. The prevalence if diabetes type 2 in a group of chronically ill hospitalised patients with schizophrenia is significantly lower than in outpatient populations. Residing in a facility long term may serve as a protective factor. The exclusion of psychological stressors such as isolation, poverty and abandonment may account for the finding.


An interactive computer program can effectively educate patients about genetic testing for breast cancer susceptibility.
As genetic testing for susceptibility to breast cancer becomes more widespread, alternative methods for educating individuals prior to testing will be needed. Our objective was to compare face-to-face education and counseling by a genetic counselor with education by an interactive computer program, assessing the effects of each on knowledge of breast cancer genetics and intent to undergo genetic testing. We used a randomized, controlled trial. Seventy-two self-referred women with a first-degree relative with breast cancer received outpatient education and counseling at the Clinical Center of the National Institutes of Health (NIH). Twenty-nine received individualized counseling from a genetic counselor (counseling group), 29 received education from an interactive computer program followed by individualized counseling (computer group), and 14 were controls. Both pre- and postintervention assessment of knowledge about breast cancer genetics and intent to undergo genetic testing were measured. The control group participants correctly answered 74% of the knowledge questions; the counselor group, 92%; and the computer group, 96% (P <.0001). Unadjusted mean knowledge scores were significantly higher in the computer group than the counselor group (P =.048), but they were equivalent when adjusted for demographic differences (P = 0.34). Intent to undergo genetic testing was influenced by the interventions: preintervention, a majority in all groups (69%) indicated that they were likely (definitely and most likely) to undergo testing; after either intervention coupled with counseling, only 44% indicated that they were likely to do so (P =.0002; odds ratio = 2.8, 95% CI = 1.7-4.9). We concluded that a computer program can successfully educate patients about breast cancer susceptibility, and, along with genetic counseling, can influence patients' intentions to undergo genetic testing.


Soft corals exert no direct effects on coral reef fish assemblages.
Correlations between abundance of organisms and their habitat have often been used as a measure of the importance of particular habitat features. However, experimental manipulation of the habitat provides a more unequivocal estimate of its importance. In this study we quantified how fish communities on small patch reefs covaried with changes in benthic cover habitat features. A random sample of small patch reefs was selected and both fish abundance and habitat measures recorded. Naturally occurring patch reefs could be classed into three habitat types based on their benthic cover. Reefs dominated by massive soft corals were the most abundant (50%), followed by those dominated by rock and soft corals in equal proportions (36%), then reefs dominated by branching corals (14%). Fish assemblages differed between the reef types. Communities on soft-coral-dominated and rock/soft-coral-dominated patch reefs formed a continuum of species responses correlated with degree of soft coral cover. In contrast, branching-coral-dominated reefs were occupied by a more discrete set of species. We tested the role of soft corals in contributing to this pattern by experimentally reducing soft coral cover on patch reefs from a baseline level of ~67% to ~33% and ~6%, and monitoring the experiment over 2 years. Contrary to expectations derived from the correlative data, and in contrast with previous manipulations of hard corals, soft-coral disturbance did not generate any corresponding changes in the fish assemblage. This "negative" result indicated that the quality and heterogeneity of habitat generated by soft corals on patch reefs was indistinguishable from equivalent-sized habitat patches formed by bare rock alone. Nevertheless, because soft corals are living organisms they have the potential to generate indirect effects by interacting with other organisms such as hard corals. In the long-term, we hypothesize that biotic interactions between habitat forming organisms might affect composition of fish assemblages on patch reefs.


Emergence of dengue virus serotype 2 in Mauritania and molecular characterization of its circulation in West Africa.
The number of sporadic and epidemic dengue fever cases have reportedly been increasing in recent years in some West African countries, such as Senegal and Mali. The first epidemic of laboratory-confirmed dengue occurred in Nouakchott, the capital city of Mauritania situated in the Saharan desert, in 2014. On-site diagnosis of dengue fever was established using a rapid diagnostic test for dengue. In parallel, the presence of Aedes aegypti mosquitoes in the city was confirmed. The initial diagnosis was confirmed by RT-PCR, which showed that all samples from the 2014 dengue epidemic in Nouakchott were dengue virus serotype 2 (DENV-2). The whole genome or envelope protein gene of these strains, together with other DENV-2 strains obtained from travelers returning from West African countries to France between 2016 and 2019 (including two Mauritanian strains in 2017 and 2018), were sequenced. Phylogenetic analysis suggested a recent emergence of an epidemic strain from the cosmopolitan genotype belonging to West African cosmopolitan lineage II, which is genetically distinct from African sylvatic genotype. The origin of this DENV-2 lineage is still unknown, but our data seem to suggest a recent and rapid dispersion of the epidemic strain throughout the region. More complete genome sequences of West African DENV-2 are required for a better understanding of the dynamics of its circulation. Arboviral surveillance and outbreak forecasting are urgently needed in West Africa.


The Big Data Revolution: Opportunities for Chief Nurse Executives.
Informatics competency adoption is a recognized issue across nursing roles in digital health practice settings. Further, it has been suggested that the health system's inability to reap the promised benefits of electronic health/patient records is, in part, a manifestation of inadequate development of informatics competency by chief nurse executives (CNEs) and other clinicians (Amendola 2008; Simpson 2013). This paper will focus on CNE informatics competency and nursing knowledge development as it pertains to the Big Data revolution. With the paper's aim of showing how CNEs armed with informatics competency can harness the full potential of Big Data offering new opportunities for nursing knowledge development in their clinical transformation roles as eHealth project sponsors. It is proposed that informatics-savvy CNEs are the new transformational leaders of the digital age who will have the advantage to successfully advocate for nurses in leading 21st-century health systems. Also, transformational CNEs armed with informatics competency will position nurses and the nursing profession to achieve its future vision, where nurses are perceived by patients and professionals alike as knowledge workers, providing the leadership essential for safe, quality care and demonstrating nursing's unique contributions to fiscal health through clinically relevant, evidence-based practices (McBride 2005b).


Management of headache in emergency room.
Headache is one of the most common symptoms that leads patients to the emergency room (ER) and is often related to diseases requiring prompt diagnosis and immediate treatment. This consideration brought us to consider the importance of the neurologist in improving the management of patients arriving in the ER with headache. We carried out a study for testing the degree of agreement between ER physician and neurologist using patient evaluation at headache centre (HC) as the gold standard. One hundred and seventeen patients with idiopathic (78) or symptomatic (39) headache were evaluated by the ER physician, the ER neurologist and the HC expert. The ER physician and the HC expert reached a fair agreement (Kappa=0.40); the other two pairs reached a moderate agreement (Kappa=0.57-0.60). There was no significant difference in the agreement of the three evaluators in patients with impairment of daily living activities or aged over 40. The agreement between the ER physician and the neurologist was lower (Kappa=0.58), especially in patients with their first headache episode. Based on our results, patients seen at the ER for a headache episode can be fairly successfully managed by the ER physician, except those who present a first attack, for whom neurological consultation is needed.


Effect of dietary antioxidants, training, and performance correlates on antioxidant status in competitive rowers.
The beneficial effects of exercise and a healthy diet are well documented in the general population but poorly understood in elite athletes. Previous research in subelite athletes suggests that regular training and an antioxidant-rich diet enhance antioxidant defenses but not performance. To investigate whether habitual diet and/or exercise (training status or performance) affect antioxidant status in elite athletes. Antioxidant blood biomarkers were assessed before and after a 30-min ergometer time trial in 28 male and 34 female rowers. The antioxidant blood biomarkers included ascorbic acid, uric acid, total antioxidant capacity (TAC), erythrocyte- superoxide dismutase, glutathione peroxidase (GPx), and catalase. Rowers completed a 7-d food diary and an antioxidant-intake questionnaire. Effects of diet, training, and performance on resting biomarkers were assessed with Pearson correlations, and their effect on exercise-induced changes in blood biomarkers was assessed by a method of standardization. With the exception of GPx, there were small to moderate increases with exercise for all markers. Blood resting TAC had a small correlation with total antioxidant intake (correlation .29; 90% confidence limits, ±.27), and the exercise-induced change in TAC had a trivial to small association with dietary antioxidant intake from vitamin C (standardized effect .19; ±.22), vegetables (.20; ±.23), and vitamin A (.25; ±.27). Most other dietary intakes had trivial associations with antioxidant biomarkers. Years of training had a small inverse correlation with TAC (-.32; ±.19) and a small association with the exercise-induced change in TAC (.27; ±.24). Training status correlates more strongly with antioxidant status than diet does.


[Let them speak for themselves: reports from professionals about their experiences in therapeutic residency training].
This paper deals with qualitative research conducted in the Therapeutic Residency Service in the municipality of Campina Grande, State of Paraíba, Brazil, the objective being to ascertain the difficulties encountered by the professionals in the healthcare practices defined by the aforementioned service and identify the prospects identified by them for the residents. A semi-structured interview routine was conducted with ten professionals for the data collection, along with analysis of field records and systematic observation. The main difficulties faced are of a structural nature, namely lack of human resources and/or training and social support. Social reintegration, insertion in the workplace and autonomy are the expectations singled out by the professionals. In light of the results, it can be affirmed that, in spite of the difficulties experienced, it is possible to see progress, such as the autonomy of some residents. Nevertheless, as it is a recent service, it needs to be constantly evaluated.


Identification of Differentially Expressed Genes in Different Glioblastoma Regions and Their Association with Cancer Stem Cell Development and Temozolomide Response.
The molecular heterogeneity of gene expression profiles of glioblastoma multiforme (GBM) are the most important prognostic factors for tumor recurrence and drug resistance. Thus, the aim of this study was to identify potential target genes related to temozolomide (TMZ) resistance and GBM recurrence. The genomic data of patients with GBM from The Cancer Genome Atlas (TCGA; 154 primary and 13 recurrent tumors) and a local cohort (29 primary and 4 recurrent tumors), samples from different tumor regions from a local cohort (29 tumor and 25 peritumoral regions), and Gene Expression Omnibus data (GSE84465, single-cell RNA sequencing; 3589 cells) were included in this study. Critical gene signatures were identified based an analysis of differentially expressed genes (DEGs). DEGs were further used to evaluate gene enrichment levels among primary and recurrent GBMs and different tumor regions through gene set enrichment analysis. Protein-protein interactions (PPIs) were incorporated into gene regulatory networks to identify the affected metabolic pathways. The enrichment levels of 135 genes were identified in the peritumoral regions as being risk signatures for tumor recurrence. Fourteen genes (DVL1, PRKACB, ARRB1, APC, MAPK9, CAMK2A, PRKCB, CACNA1A, ERBB4, RASGRF1, NF1, RPS6KA2, MAPK8IP2, and PPM1A) derived from the PPI network of 135 genes were upregulated and involved in the regulation of cancer stem cell (CSC) development and relevant signaling pathways (Notch, Hedgehog, Wnt, and MAPK). The single-cell data analysis results indicated that 14 key genes were mainly expressed in oligodendrocyte progenitor cells, which could produce a CSC niche in the peritumoral region. The enrichment levels of 336 genes were identified as biomarkers for evaluating TMZ resistance in the solid tumor region. Eleven genes (ARID5A, CDC42EP3, CDKN1A, FLT3, JUNB, MAP2K3, MYBPC2, RGS14, RNASEK, TBC1D30, and TXNDC11) derived from the PPI network of 336 genes were upregulated and may be associated with a high risk of TMZ resistance; these genes were identified in both the TCGA and local cohorts. Furthermore, the expression patterns of ARID5A, CDKN1A, and MAP2K3 were identical to the gene signatures of TMZ-resistant cell lines. The identified enrichment levels of the two gene sets expressed in tumor and peritumoral regions are potentially helpful for evaluating TMZ resistance in GBM. Moreover, these key genes could be used as biomarkers, potentially providing new molecular strategies for GBM treatment.


Association of HbA1c levels with vascular complications and death in patients with type 2 diabetes: evidence of glycaemic thresholds.
There is conflicting evidence regarding appropriate glycaemic targets for patients with type 2 diabetes. Here, we investigate the relationship between HbA(1c) and the risks of vascular complications and death in such patients. Eleven thousand one hundred and forty patients were randomised to intensive or standard glucose control in the Action in Diabetes and Vascular disease: Preterax and Diamicron Modified Release Controlled Evaluation (ADVANCE) trial. Glycaemic exposure was assessed as the mean of HbA(1c) measurements during follow-up and prior to the first event. Adjusted risks for each HbA(1c) decile were estimated using Cox models. Possible differences in the association between HbA(1c) and risks at different levels of HbA(1c) were explored using linear spline models. There was a non-linear relationship between mean HbA(1c) during follow-up and the risks of macrovascular events, microvascular events and death. Within the range of HbA(1c) studied (5.5-10.5%), there was evidence of 'thresholds', such that below HbA(1c) levels of 7.0% for macrovascular events and death, and 6.5% for microvascular events, there was no significant change in risks (all p > 0.8). Above these thresholds, the risks increased significantly: every 1% higher HbA(1c) level was associated with a 38% higher risk of a macrovascular event, a 40% higher risk of a microvascular event and a 38% higher risk of death (all p < 0.0001). In patients with type 2 diabetes, HbA(1c) levels were associated with lower risks of macrovascular events and death down to a threshold of 7.0% and microvascular events down to a threshold of 6.5%. There was no evidence of lower risks below these levels but neither was there clear evidence of harm.


Functional coherence of the human Y chromosome.
A systematic search of the nonrecombining region of the human Y chromosome (NRY) identified 12 novel genes or families, 10 with full-length complementary DNA sequences. All 12 genes, and six of eight NRY genes or families previously isolated by less systematic means, fell into two classes. Genes in the first group were expressed in many organs; these housekeeping genes have X homologs that escape X inactivation. The second group, consisting of Y-chromosomal gene families expressed specifically in testes, may account for infertility among men with Y deletions. The coherence of the NRY's gene content contrasts with the apparently haphazard content of most eukaryotic chromosomes.


Regulation of the glyoxylate bypass operon: cloning and characterization of iclR.
In Escherichia coli, expression of the glyoxylate bypass operon appears to be controlled, in part, by the product of iclR+. Mutations in iclR have been found to yield constitutive expression of this operon, suggesting that iclR+ encodes a repressor protein. We have cloned iclR+ by taking advantage of its tight genetic linkage with the glyoxylate bypass operon. The clone complemented a mutant allele of iclR in trans, restoring an inducible phenotype for this operon. Deletion analysis identified a region of ca. 900 base pairs that was necessary and sufficient for complementation. The nucleotide sequence of the insert was then determined. Translation of this sequence revealed an open reading frame capable of encoding a protein with Mr 29,741 preceded by a potential Shine-Dalgarno ribosome-binding site. The deduced amino acid sequence includes a region at the amino terminus that may form a helix-turn-helix motif, a structure found in many DNA-binding domains.


Enteric and nasal shedding of bovine torovirus (Breda virus) in feedlot cattle.
To assess fecal and nasal shedding patterns of bovine torovirus (BoTV) in cattle at time of arrival and periodically throughout the first 21 days after arrival at a feedlot. 57 steers. Fecal and nasal-swab samples collected on days 0, 4, 14, and 21 after arrival were tested for BoTV, using ELISA. A subset of samples from calves testing positive and negative for BoTV was analyzed, using reverse transcriptase-polymerase chain reaction (RT-PCR). Paired serum samples were collected on days 0 and 21 and tested for BoTV antibodies, using a hemagglutination inhibition assay. Overall rate of fecal shedding of BoTV was 21 of 57 (37%) by ELISA and 40 of 42 (95%) by RT-PCR with peak shedding on day 4. Diarrhea was more common in calves shedding BoTV than those not shedding the virus (odds ratio, 1.72). Overall rate of nasal shedding of BoTV was 15 of 57 (26%) by ELISA and 42 of 42 (100%) by RT-PCR, with peak shedding on day 0. Specificity of the RT-PCR product was confirmed by sequence analysis. Approximately 93% of the calves seroconverted to BoTV (> 4-fold increase in titer). Differences were not detected between calves shedding BoTV and nonshedders in relation to disease and treatments, perhaps because of the low number of cattle in the study. This study confirmed BoTV infections in feedlot cattle, including BoTV antigen and viral RNA in nasal secretions, and the shedding pattern during the first 21 days after arrival in a feedlot.


Endoplasmic reticulum stress regulates the innate immunity critical transcription factor IRF3.
IFN regulatory factor 3 (IRF3) regulates early type I IFNs and other genes involved in innate immunity. We have previously shown that cells undergoing an endoplasmic reticulum (ER) stress response called the unfolded protein response produce synergistically augmented IFN-β when stimulated with pattern recognition receptor agonists such as LPS. Concomitant ER stress and LPS stimulation resulted in greater recruitment of the IRF3 transcription factor to ifnb1 gene regulatory elements. In this study, we used murine cells to demonstrate that both oxygen-glucose deprivation and pharmacologic unfolded protein response inducers trigger phosphorylation and nuclear translocation of IRF3, even in the absence of exogenous LPS. Different ER stressors used distinct mechanisms to activate IRF3: IRF3 phosphorylation due to calcium-mobilizing ER stress (thapsigargin treatment, oxygen-glucose deprivation) critically depended upon stimulator of IFN gene, an ER-resident nucleic acid-responsive molecule. However, calcium mobilization alone by ionomycin was insufficient for IRF3 phosphorylation. In contrast, other forms of ER stress (e.g., tunicamycin treatment) promote IRF3 phosphorylation independently of stimulator of IFN gene and TANK-binding kinase 1. Rather, IRF3 activation by tunicamycin and 2-deoxyglucose was inhibited by 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride, a serine protease inhibitor that blocks activating transcription factor 6 processing. Interfering with ER stress-induced IRF3 activation abrogated IFN-β synergy. Together, these data suggest ER stress primes cells to respond to innate immune stimuli by activating the IRF3 transcription factor. Our results also suggest certain types of ER stress accomplish IRF3 phosphorylation by co-opting existing innate immune pathogen response pathways. These data have implications for diseases involving ER stress and type I IFN.


Effect of using proper light-curing techniques on energy delivered to a Class 1 restoration.
To determine the effect of proper light-curing instruction on the radiant exposure (energy density) delivered by dentists using six dental curing lights to a posterior Class 1 restoration. Twenty-five dentists attending a professional meeting were instructed to position a patient simulator (MARC-PS, BlueLight), as they would for a patient, and then to expose the simulated Class 1 maxillary second molar preparation for a specified amount of time. At this point, the dentists were unaware of the purpose of the experiment. Each participant used three different curing lights, and the irradiance and radiant exposure (J/cm2) delivered to the preparation was recorded. Participants were then informed of the purpose of the exercise, and given specific light-curing instructions and training using the patient simulator, after which they re-exposed the same preparation using the same curing lights. Pre- and post-instruction radiant exposure values were compared using one-way ANOVA (α = .05), and for each light among all operators using a two-tailed, paired Student's t test. There was a wide variation in the radiant exposure delivered by the dentists and by the six curing lights, from 2.9 to 15.4 J/cm2. Before receiving additional light-curing instruction, 68% of dentists delivered less than 10 J/cm2. The radiant exposure delivered increased significantly (P < .001) by up to 30%, as a result of training using MARC-PS. The results indicate that some of the dentists participating in the present study delivered an inadequate amount of radiant exposure before instruction. More energy was delivered after a short training session using the MARC-PS. Reinforcing the proper photo-curing techniques may improve the outcome when placing resin-based restorations.


Layer-specific expression of extracellular matrix molecules in the mouse somatosensory and piriform cortices.
In the developing central nervous system (CNS), extracellular matrix (ECM) molecules have regulating roles such as in brain development, neural-circuit maturation, and synaptic-function control. However, excluding the perineuronal net (PNN) area, the distribution, constituent elements, and expression level of granular ECM molecules (diffuse ECM) present in the mature CNS remain unclear. Diffuse ECM molecules in the CNS share the components of PNNs and are likely functional. As cortical functions are greatly region-dependent, we hypothesized that ECM molecules would differ in distribution, expression level, and components in a region- and layer-dependent manner. We examined the layer-specific expression of several chondroitin sulfate proteoglycans (aggrecan, neurocan, and brevican), tenascin-R, Wisteria floribunda agglutinin (WFA)-positive molecules, hyaluronic acid, and link protein in the somatosensory and piriform cortices of mature mice. Furthermore, we investigated expression changes in WFA-positive molecules due to aging. In the somatosensory cortex, PNN density was particularly high at layer 4 (L4), but not all diffuse ECM molecules were highly expressed at L4 compared to the other layers. There was almost no change in tenascin-R and hyaluronic acid in any somatosensory-cortex layer. Neurocan showed high expression in L1 of the somatosensory cortex. In the piriform cortex, many ECM molecules showed higher expression in L1 than in the other layers. However, hyaluronic acid showed high expression in deep layers. Here, we clarified that ECM molecules differ in constituent elements and expression in a region- and layer-dependent manner. Region-specific expression of ECM molecules is possibly related to functions such as region-specific plasticity and vulnerability.


Development and validation of a method for quantification of two tobacco-specific nitrosamines in indoor air.
A sensitive and accurate method for the quantification of 1'-Demethyl-1'-nitrosonicotine (NNN) and 4-(methylnitrosamino)-1-(3-Pyridyl)-1-butanone (NNK) in indoor air was developed and validated. To this aim, a novel approach for the collection of two tobacco-specific nitrosamines, using silica sorbent cartridges followed by simplified sample preparation and isotope dilution liquid chromatography/electrospray ionization tandem mass spectrometry, was applied. This procedure led to a substantial improvement in terms of sensitivity and sample throughput as compared with methods using conventional trapping. For the validation, a matrix-based approach using an accuracy profile procedure was selected. The evaluated matrices were background air samples, environmental aerosols of a heat-not-burn tobacco product (Tobacco Heating System [THS] 2.2, commercialized under the brand IQOS®), a rechargeable electronic cigarette (Solaris®), and the environmental tobacco smoke (ETS) of a conventional cigarette (Marlboro Gold®). The method showed excellent recoveries, sensitivity, and precision. The limits of detection of the method for NNN and NNK were 0.0108 ng/m3 and 0.0136 ng/m3, respectively. The calibration range of the instrument spanned 0.2-60 ng/mL. The calculated lower working range limit (LWRL) of the method for NNN was 0.126 ng/m3, and the LWRL for NNK was 0.195 ng/m3. The method was applied to evaluate surrogate environmental aerosols generated using smoking machines. This model is reliable but gives a large overestimation of the possible impact of THS 2.2 and e-cigarettes on indoor air, because the retention of NNN and NNK in the body of the consumers is not taken into account. As a consequence, the values reported do not reflect a real-life setting. The contents of the two target compounds in the surrogate environmental aerosols were 0.0830 ± 0.0153 ng/m3 of NNN and 0.0653 ± 0.0138 ng/m3 of NNK for THS 2.2, 0.0561 ± 0.0296 ng/m3 of NNN for e-cigarettes, and 0.816 ± 0.109 ng/m3 of NNN and 4.13 ± 1.04 ng/m3 NNK for cigarettes. These values correspond to 10% of the measured ETS concentration for NNN in environmental aerosols of THS 2.2 and 7% for those of e-cigarettes. For NNK, the value for the environmental aerosol of THS 2.2 was 2% of the ETS value.


Involvement of the retinoblastoma protein in monocytic and neutrophilic lineage commitment of human bone marrow progenitor cells.
The retinoblastoma gene product (pRb) is involved in both cell cycle regulation and cell differentiation. pRb may have dual functions during cell differentiation: partly by promoting a cell cycle brake at G(1) and also by interacting with tissue-specific transcription factors. We recently showed that pRb mediates differentiation of leukemic cell lines involving mechanisms other than the induction of G(1) arrest. In the present study, we investigated the role of pRb in differentiation of human bone marrow progenitor cells. Human bone marrow cells were cultured in a colony-forming unit-granulocyte-macrophage (CFU-GM) assay. The addition of antisense RB oligonucleotides (alpha-RB), but not the addition of sense orientated oligonucleotides (SO) or scrambled oligonucleotides (SCR), reduced the number of colonies staining for nonspecific esterase without affecting the clonogenic growth. Monocytic differentiation of CD34(+) cells supported by FLT3-ligand and interleukin-3 (IL-3) was correlated to high levels of hypophosphorylated pRb, whereas neutrophilic differentiation, supported by granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF), was correlated to low levels. The addition of alpha-RB to liquid cultures of CD34(+) cells, supported with FLT3-ligand and IL-3, inhibited monocytic differentiation. This was judged by morphology, the expression of CD14, and staining for esterase. Moreover, the inhibition of monocytic differentiation of CD34(+) cells mediated by alpha-RB, which is capable of reducing pRb expression, was counterbalanced by an enhanced neutrophilic differentiation response, as judged by morphology and the expression of lactoferrin. CD34(+) cells incubated with oligo buffer, alpha-RB, SO, or SCR showed similar growth rates. Taken together, these data suggest that pRb plays a critical role in the monocytic and neutrophilic lineage commitment of human bone marrow progenitors, probably by mechanisms that are not strictly related to control of cell cycle progression.


True single-molecule DNA sequencing of a pleistocene horse bone.
Second-generation sequencing platforms have revolutionized the field of ancient DNA, opening access to complete genomes of past individuals and extinct species. However, these platforms are dependent on library construction and amplification steps that may result in sequences that do not reflect the original DNA template composition. This is particularly true for ancient DNA, where templates have undergone extensive damage post-mortem. Here, we report the results of the first "true single molecule sequencing" of ancient DNA. We generated 115.9 Mb and 76.9 Mb of DNA sequences from a permafrost-preserved Pleistocene horse bone using the Helicos HeliScope and Illumina GAIIx platforms, respectively. We find that the percentage of endogenous DNA sequences derived from the horse is higher among the Helicos data than Illumina data. This result indicates that the molecular biology tools used to generate sequencing libraries of ancient DNA molecules, as required for second-generation sequencing, introduce biases into the data that reduce the efficiency of the sequencing process and limit our ability to fully explore the molecular complexity of ancient DNA extracts. We demonstrate that simple modifications to the standard Helicos DNA template preparation protocol further increase the proportion of horse DNA for this sample by threefold. Comparison of Helicos-specific biases and sequence errors in modern DNA with those in ancient DNA also reveals extensive cytosine deamination damage at the 3' ends of ancient templates, indicating the presence of 3'-sequence overhangs. Our results suggest that paleogenomes could be sequenced in an unprecedented manner by combining current second- and third-generation sequencing approaches.


Should asymptomatic men be included in chlamydia screening programs? Cost-effectiveness of chlamydia screening among male and female entrants to a national job training program.
To compare the cost-effectiveness of various chlamydia screening strategies within a population of male and female youth entering a national job training program. Cost-effectiveness analysis of various chlamydia screening strategies among a cohort of 4000 female and male New England job training students. Strategies for women include (a) no screening, (b) universal endocervical DNA probe screening, (c) universal urine based NAAT screening, and (d) universal endocervical NAAT screening. Strategies for men include (a) no screening, (b) selective urine NAAT screening of leukocyte esterase (LE)-positive urines, and (c) universal urine-based NAAT screening. Universal endocervical NAAT screening of women and universal urine NAAT screening of men were the most effective and cost-effective strategies individually and in combination. Endocervical NAAT screening of women prevented 23 more cases of PID and saved $27,000 more than endocervical DNA probe screening. Likewise, universal urine NAAT screening of men prevented 21 more cases of PID in their female partners and saved $16,000 more than selective urine NAAT screening of LE positive men. Use of a sensitive NAAT to screen both men and women for chlamydia upon entry to a National Job Training Program is cost-effective, cost-saving, and provides a public health opportunity to substantially reduce chlamydia infections among youth at risk for sexually transmitted diseases.


Immune cells as targets for cardioprotection: new players and novel therapeutic opportunities.
New therapies are required to reduce myocardial infarct (MI) size and prevent the onset of heart failure in patients presenting with acute myocardial infarction (AMI), one of the leading causes of death and disability globally. In this regard, the immune cell response to AMI, which comprises an initial pro-inflammatory reaction followed by an anti-inflammatory phase, contributes to final MI size and post-AMI remodelling [changes in left ventricular (LV) size and function]. The transition between these two phases is critical in this regard, with a persistent and severe pro-inflammatory reaction leading to adverse LV remodelling and increased propensity for developing heart failure. In this review article, we provide an overview of the immune cells involved in orchestrating the complex and dynamic inflammatory response to AMI-these include neutrophils, monocytes/macrophages, and emerging players such as dendritic cells, lymphocytes, pericardial lymphoid cells, endothelial cells, and cardiac fibroblasts. We discuss potential reasons for past failures of anti-inflammatory cardioprotective therapies, and highlight new treatment targets for modulating the immune cell response to AMI, as a potential therapeutic strategy to improve clinical outcomes in AMI patients. This article is part of a Cardiovascular Research Spotlight Issue entitled 'Cardioprotection Beyond the Cardiomyocyte', and emerged as part of the discussions of the European Union (EU)-CARDIOPROTECTION Cooperation in Science and Technology (COST) Action, CA16225.


Risk for perinatal HIV-1 transmission according to maternal immunologic, virologic, and placental factors.
To evaluate how maternal and obstetric factors interact to influence mother-to-child human immunodeficiency virus type 1 (HIV-1) transmission. Prospective, observational cohort study of children born to HIV-infected women to determine child's HIV infection status. The analysis then compared peripartum maternal, placental, and obstetric variables between HIV-1 transmitter and nontransmitter women. Two large maternity wards in Kinshasa, Zaire. Consecutive sample of 324 HIV-1-infected women at delivery, with 254 HIV-seronegative women followed up as control subjects. PRINCIPAL OUTCOME MEASURES: HIV infection status of children, to classify each woman as an HIV-1 transmitter or nontransmitter. The highest transmission risk (TR) was associated with maternal p24 antigenemia (TR, 71%; relative risk [RR], 3.0; 95% confidence interval [CI], 1.7 to 5.2) and maternal CD8+ lymphocyte counts of at least 1.80 x 10(9)/L (1800/microL) (TR, 50%; RR, 2.2; 95% CI, 1.2 to 4.2). Among women with CD8+ lymphocyte counts of less than 1.80 x 10(9)/L, CD4+ lymphocyte counts of less than 0.60 x 10(9)/L were a risk factor (TR, 29%; RR, 2.2; 95% CI, 1.2 to 4.2). In women with neither high CD8+ nor low CD4+ lymphocyte counts, placental membrane inflammation was associated with perinatal transmission (TR, 40%; RR, 4.2; 95% CI, 1.3 to 13.7). In women with neither p24 antigenemia, high CD8+ or low CD4+ lymphocyte counts, nor placental membrane inflammation, the transmission risk was only 7%. Additional correlates of transmission included maternal anemia and fever, but not maternal sexually transmitted diseases. Identifiable subgroups of HIV-1-infected women based on maternal and placental characteristics had between a 7% and 71% risk of perinatal HIV-1 transmission. Not only the overall rate of transmission but the impact of different risk factors for transmission appear to vary over the course of HIV infection.


Total arch replacement under flow monitoring during selective cerebral perfusion using a single pump.
Flow in individual vessels is passively determined when a single pump is used for selective cerebral perfusion during aortic arch surgery. We installed a Doppler flowmeter in the circuit and measured flow in the supraaortic vessels to determine flow distribution during selective cerebral perfusion. We cannulated and perfused three supraaortic vessels using a single pump in 203 patients who underwent elective (n = 158) or emergency or urgent (n = 45) total arch replacement using a four-branched prosthetic graft. Flow rates in each branch were continuously monitored during selective cerebral perfusion. The respective mean flow rates in the brachiocephalic, left common carotid, and left subclavian arteries and total flow rates were 5.8, 3.3, 3.4, and 12.5 mL·kg(-1)·min(-1). The ratios of flow in these vessels to total flow were 46.5%, 26.5%, and 27.0%, respectively, and they were not affected by the total flow rate. In-hospital mortality rates among the patients who underwent elective and emergency or urgent surgery were 1.9% (n = 3) and 11.1% (n = 5), respectively, and the rates of postoperative stroke were 2.5% (n = 4) and 8.9% (n = 4), respectively. Total flow in the supraaortic vessels during selective cerebral perfusion was significantly lower in patients with neurologic complications than in those without (732 versus 806 mL/min; p = 0.034). Flow monitoring showed that selective perfusion using a single pump adequately distributed flow among all supraaortic vessels. This monitoring system might help to improve brain protection and outcomes during total aortic arch replacement.


Exocytosis of vacuolar apical compartment (VAC): a cell-cell contact controlled mechanism for the establishment of the apical plasma membrane domain in epithelial cells.
The vacuolar apical compartment (VAC) is an organelle found in Madin-Darby canine kidney (MDCK) cells with incomplete intercellular contacts by incubation in 5 microM Ca++ and in cells without contacts (single cells in subconfluent culture); characteristically, it displays apical biochemical markers and microvilli and excludes basolateral markers (Vega-Salas, D. E., P. J. I. Salas, and E. Rodriguez-Boulan. 1987. J. Cell Biol. 104:1249-1259). The apical surface of cells kept under these culture conditions is immature, with reduced numbers of microvilli and decreased levels of an apical biochemical marker (184 kD), which is, however, still highly polarized (Vega-Salas, D. E., P. J. I. Salas, D. Gundersen, and E. Rodriguez-Boulan. 1987. J. Cell Biol. 104:905-916). We describe here the morphological stages of VAC exocytosis which ultimately lead to the establishment of a differentiated apical domain. Addition of 1.8 mM Ca++ to monolayers developed in 5 microM Ca++ causes the rapid (20-40 min) fusion of VACs with the plasma membrane and their accessibility to external antibodies, as demonstrated by immunofluorescence, immunoperoxidase EM, and RIA with antibodies against the 184-kD apical plasma membrane marker. Exocytosis occurs towards areas of cell-cell contact in the developing lateral surface where they form intercellular pockets; fusion images are always observed immediately adjacent to the incomplete junctional bands detected by the ZO-1 antibody (Stevenson, B. R., J. D. Siliciano, M. S. Mooseker, and D. A. Goodenough. 1986. J. Cell Biol. 103:755-766). Blocks of newly incorporated VAC microvilli and 184-kD protein progressively move from intercellular ("primitive" lateral) spaces towards the microvilli-poor free cell surface. The definitive lateral domain is sealed behind these blocks by the growing tight junctional fence. These results demonstrate a fundamental role of cell-cell contact-mediated VAC exocytosis in the establishment of epithelial surface polarity. Because isolated stages (intercellular pockets) of the stereotyped sequence of events triggered by the establishment of intercellular contacts in MDCK cells have been reported during normal differentiation of intestine epithelium (Colony, P. C., and M. R. Neutra. 1983. Dev. Biol. 97:349-363), we speculate that the mechanism we describe here plays an important role in the establishment of epithelial cell polarity in vivo.


Effects of chronic systemic administration of basic fibroblast growth factor on collateral development in the canine heart.
Recently we reported that intracoronary administration of basic fibroblast growth factor (bFGF), a potent angiogenic peptide, increases collateral blood flow in dogs subjected to progressive left circumflex coronary artery (LCx) occlusion. The aim of the present study was to examine the effect of systemically administered bFGF on collateral blood flow and to assess its pharmacokinetics and potential side effects. Forty-seven dogs were subjected to progressive ameroid-induced occlusion of the LCx, an intervention known to induce the development of collateral vessels. In phase I of the investigation, dogs were randomized to receive bFGF 1.74 mg/d (n = 10) or saline (n = 9) as a left atrial injection for 4 weeks. Relative collateral blood flow was assessed serially with radiolabeled microspheres in the conscious state during maximal coronary vasodilatation. Initiation of bFGF treatment was temporally associated with a marked acceleration of collateral development; however, collateral flow in control dogs improved toward the end of the study, approaching that of bFGF-treated dogs at the 38-day end point. Phase II of the investigation was a three-armed study of extended duration to determine whether bFGF caused a sustained increase in collateral function. Dogs were randomized to receive bFGF 1.74 mg/d for 9 weeks (n = 7), bFGF 1.74 mg/d for 5 weeks followed by placebo for 4 weeks (n = 11), or placebo for 9 weeks (n = 10). Relative and absolute collateral blood flow were assessed serially with microspheres during maximal coronary vasodilatation. Between the 10th and 17th days after ameroid placement, bFGF-treated dogs exhibited marked improvement in collateral flow such that maximal collateral conductance exceeded that of controls by 24% at the 5-week crossover point. Final collateral conductance was similar in dogs receiving bFGF for 5 and 9 weeks despite withdrawal of treatment in the former group. bFGF administration was associated with a 21% increase in final collateral conductance as well as a 49% increase in collateral zone vascular density. Prolonged bFGF administration was also associated with a decrease in arterial pressure, moderate thrombocytopenia, and moderate, reversible anemia. Systemic administration of bFGF enhanced collateral conductance in dogs with progressive single-vessel coronary occlusion. The beneficial effect of bFGF occurred primarily between the 7th and 14th days of therapy, and regression of collateral development was not noted after withdrawal of treatment. The present investigation provides impetus to the concept that collateral development can be enhanced pharmacologically-specifically by bFGF-raising the possibility that such an intervention might eventually be applied clinically.


Thyroid nodules with nondiagnostic results on repeat fine-needle aspiration biopsy: which nodules should be considered for repeat biopsy or surgery rather than follow-up?
The goal of this study was to assess the clinicopathologic and ultrasonographic features of thyroid nodules with nondiagnostic results on repeat ultrasonography (US)-guided fineneedle aspiration biopsy (FNAB) according to size and the number of suspicious findings and to determine the proper management of nodules with consecutive nondiagnostic results. This retrospective study included 297 nodules with nondiagnostic results on repeat FNAB that were evaluated by US over the course of at least 12 months of follow-up, a follow-up biopsy, or an operation. We compared clinical and US variables between benign and malignant nodules in thyroid nodules with repeat nondiagnostic results. The comparison of benign and malignant nodules with repeat nondiagnostic results revealed that age, marked hypoechogenicity, irregular or microlobulated margins, microcalcifications, and nonparallel shape were significantly associated with malignancy. Multivariate logistic regression analysis in malignant nodules revealed that microcalcifications and irregular or microlobulated margins were independently associated with malignancy. Among them, only irregular or microlobulated margins were independently significant as a predictor of malignancy in repeatedly nondiagnostic nodules measuring >10 mm. Using receiver operating characteristic analysis, the best cutoff value for the "number of suspicious findings" between benign and malignant nodules was three in nodules of all sizes, three in nodules measuring ≤10 mm, and two in nodules measuring >10 mm. Irregular or microlobulated margins may be the most frequent US features in repeatedly nondiagnostic nodules >10 mm. The presence of "two or more suspicious findings" can be used as the cutoff for distinguishing benign and malignant nodules.


Effects of kelp supplementation on thyroid function in euthyroid subjects.
To study the effects of ingestion of two different doses of supplemental kelp on the thyroid function of healthy euthyroid subjects. We conducted a double-blind prospective clinical trial involving 36 healthy euthyroid subjects, who were randomly assigned to receive placebo (4 alfalfa capsules per day), low-dose kelp (2 kelp capsules and 2 alfalfa capsules per day), or high-dose kelp (4 kelp capsules per day) for 4 weeks. Thyrotropin (thyroid-stimulating hormone or TSH), free thyroxine, and total triiodothyronine were assessed at weeks 0, 4, and 6. Response to thyrotropin-releasing hormone stimulation, urinary iodine excretion, and basal metabolic rate were determined at weeks 0 and 4. TSH concentrations did not differ significantly between week 0 and week 4 in the placebo group (P = 0.16) but increased significantly in both the low-dose kelp (P = 0.04) and high-dose kelp (P = 0.002) groups. Free thyroxine concentrations decreased slightly but significantly after 4 weeks of placebo but were unchanged in the low-dose and the high-dose kelp groups. In contrast, total triiodothyronine levels did not differ significantly after 4 weeks of placebo or low-dose kelp therapy but were significantly decreased after high-dose kelp therapy (P = 0.04). Similarly, the thyrotropin-releasing hormone stimulation test showed no significant change in poststimulation TSH after 4 weeks in the placebo or low-dose kelp groups but revealed a significantly increased response after high-dose kelp therapy (P = 0.0002). The 24-hour urinary iodine excretion showed dose-dependent increases in the two kelp study groups. Basal metabolic rate did not change significantly in any study group during the 4-week study period. All thyroid laboratory values returned to baseline 2 weeks after cessation of kelp supplementation, except for TSH in the high-dose kelp group, which was significantly decreased. Short-term dietary supplementation with kelp significantly increases both basal and poststimulation TSH. These findings corroborate previous studies on the effects of supplemental iodide given to euthyroid subjects for a similar period. Further studies are needed to determine whether long-term kelp supplementation would cause clinically significant thyroid disease.


Selective chemoprecipitation and subsequent release of tagged species for the analysis of nitropeptides by liquid chromatography-tandem mass spectrometry.
Tyrosine nitration is a low-abundance post-translational protein modification that requires appropriate enrichment techniques to enable proteomic analyses. We report a simple yet highly specific method to enrich nitropeptides by chemoprecipitation involving only two straightforward chemical modifications of the nitropeptides before capturing the obtained derivatives with a strategically designed solid-phase active ester reagent. Specifically, capping of the aliphatic amines in the peptides is done first by reductive methylation to preserve the charge state of peptides for electrospray ionization mass spectrometric analysis, followed by reduction of nitrotyrosines to the corresponding aminotyrosines. These peptides are then immobilized on the solid-phase active ester reagent, whereas other peptides carrying no free amino groups are separated from the immobilized species by thoroughly washing the beads from which the tagged peptide derivatives can easily be released by acid-catalyzed hydrolysis at room temperature. The benefits of selective enrichment from a matrix of unmodified peptides for liquid chromatography-tandem mass spectrometry are demonstrated on three synthetic nitropeptides that are nitrated fragments of biologically relevant proteins. Identification of several in vitro nitrated human plasma proteins, also implicated under various pathological processes, by database searches from the enriched and tagged tryptic nitropeptides is presented as a practical application. We also show that converting the nitro-group to the small 4-formylbenzoylamido tag does not significantly alter fragmentation properties upon collision-induced dissociation compared with those of the native nitropeptides, and at the same time this derivatization actually improves electron capture dissociation due to conversion of the electron-predator nitro-group to this novel tag.


Electrocatalytic synthesis of ammonia by surface proton hopping.
Highly efficient ammonia synthesis at a low temperature is desirable for future energy and material sources. We accomplished efficient electrocatalytic low-temperature ammonia synthesis with the highest yield ever reported. The maximum ammonia synthesis rate was 30 099 μmol gcat-1 h-1 over a 9.9 wt% Cs/5.0 wt% Ru/SrZrO3 catalyst, which is a very high rate. Proton hopping on the surface of the heterogeneous catalyst played an important role in the reaction, revealed by in situ IR measurements. Hopping protons activate N2 even at low temperatures, and they moderate the harsh reaction condition requirements. Application of an electric field to the catalyst resulted in a drastic decrease in the apparent activation energy from 121 kJ mol-1 to 37 kJ mol-1. N2 dissociative adsorption is markedly promoted by the application of the electric field, as evidenced by DFT calculations. The process described herein opens the door for small-scale, on-demand ammonia synthesis.


Long-range ordering of highly charged self-assembled nanofilaments.
Charged nanoscale filaments are well-known in natural systems such as filamentous viruses and the cellular cytoskeleton. The unique properties of these structures have inspired the design of self-assembled nanofibers for applications in regenerative medicine, drug delivery, and catalysis, among others. We report here on an amphiphile of completely different chemistry based on azobenzene and a quaternary ammonium bromide headgroup that self-assembles into highly charged nanofibers in water and orders into two-dimensional crystals. Interestingly small-angle X-ray scattering (SAXS) shows that these fibers of 5.6 nm cross-sectional diameter order into crystalline arrays with remarkably large interfiber spacings of up to 130 nm. Solution concentration and temperature can be adjusted to control the interfiber spacings, and addition of salt destroyed the crystal packing indicating the electrostatic repulsions are necessary for the observed ordering. Our findings here demonstrate the universal nature of this phenomenon in systems of highly charged nanoscale filaments.


Prdm6 drives ductus arteriosus closure by promoting ductus arteriosus smooth muscle cell identity and contractility.
Based upon our demonstration that the smooth muscle cell-selective (SMC-selective) putative methyltransferase, Prdm6, interacts with myocardin-related transcription factor-A, we examined Prdm6's role in SMCs in vivo using cell type-specific knockout mouse models. Although SMC-specific depletion of Prdm6 in adult mice was well tolerated, Prdm6 depletion in Wnt1-expressing cells during development resulted in perinatal lethality and a completely penetrant patent ductus arteriosus (DA) phenotype. Lineage tracing experiments in Wnt1Cre2 Prdm6fl/fl ROSA26LacZ mice revealed normal neural crest-derived SMC investment of the outflow tract. In contrast, myography measurements on DA segments isolated from E18.5 embryos indicated that Prdm6 depletion significantly reduced DA tone and contractility. RNA-Seq analyses on DA and ascending aorta samples at E18.5 identified a DA-enriched gene program that included many SMC-selective contractile associated proteins that was downregulated by Prdm6 depletion. Chromatin immunoprecipitation-sequencing experiments in outflow tract SMCs demonstrated that 50% of the genes Prdm6 depletion altered contained Prdm6 binding sites. Finally, using several genome-wide data sets, we identified an SMC-selective enhancer within the Prdm6 third intron that exhibited allele-specific activity, providing evidence that rs17149944 may be the causal SNP for a cardiovascular disease GWAS locus identified within the human PRDM6 gene.


Protozoan parasites of the intestinal tract: a review of Coccidia and Microsporida.
Ubiquitous in nature, members of the Coccidia and Microsporida are being reported with increasing frequency in the immunocompromised as well as the immunocompetent population. These protozoans are primarily waterborne, but foodborne disease has also been reported. These organisms are responsible for acute, as well as protracted, cases of watery diarrhea with various other related sequelae. The Coccidia includes three genera--Cryptospridium, Isospora, and Cyclospora. The latter two are of lesser importance in terms of morbidity and mortality. The Microsporida includes genera (Enterocytozoon, Encephalitozoon) only recently recognized as important agents of disease. Unlike the Coccidia, these organisms are more restricted to the immunocompromised population. Increased incidence and numbers of patients with prolonged diarrhea due to these forms indicate the need for increased clinical vigilance with regard to prevention, diagnosis, and treatment.


Visual discrimination learning in the water maze: a novel test for visual acuity.
Learning about space, the environment and specific objects comprising three-dimensional arrangements requires processing of visual information. As learning and memory experiments in mammals rely heavily on normal processing of visual cues, drug-induced disruption of acquisition learning or memory formation necessitates the important control for visual acuity. A popular task used frequently for rats is the Morris water maze. However, previously used visual tasks in the water maze only control for gross visual disturbances. Here we describe a new training procedure enabling visual acuity to be tested in the water maze. Animals were trained to discriminate between two cue cards containing a pattern of vertical black and white stripes. Cards were presented in two adjacent quadrants separated by a barrier with the escape platform located in front of the smaller stripes (1 cm wide). Once 80% correct responses were attained, the wider cue card (normally 5 cm wide stripes) was randomly changed to gratings of 1,2,3,4,5, and 10 cm width. Animals learned the discrimination with acuity of 1.5 c/deg. A detailed analysis of the swim patterns further suggests that, independent of the grating used, animals make a choice immediately after release and swim along the walls towards the cue. In a further acuity test taken a few weeks later when animals were given saline infusions, performance was better than in the first test suggesting an effect of learning. This novel test may prove useful in determining subtle drug-induced deficits in visual acuity that may contribute to disruption of spatial performance in the water maze.


Differences in the fungal communities nursed by two genetic groups of the alpine cushion plant, Silene acaulis.
Foundation plants shape the composition of local biotic communities and abiotic environments, but the impact of a plant's intraspecific variations on these processes is poorly understood. We examined these links in the alpine cushion moss campion (Silene acaulis) on two neighboring mountain ranges in the French Alps. Genotyping of cushion plants revealed two genetic clusters matching known subspecies. The exscapa subspecies was found on both limestone and granite, while the longiscapa one was only found on limestone. Even on similar limestone bedrock, cushion soils from the two S. acaulis subspecies deeply differed in their impact on soil abiotic conditions. They further strikingly differed from each other and from the surrounding bare soils in fungal community composition. Plant genotype variations accounted for a large part of the fungal composition variability in cushion soils, even when considering geography or soil chemistry, and particularly for the dominant molecular operational taxonomic units (MOTUs). Both saprophytic and biotrophic fungal taxa were related to the MOTUs recurrently associated with a single plant genetic cluster. Moreover, the putative phytopathogens were abundant, and within the same genus (Cladosporium) or species (Pyrenopeziza brassicae), MOTUs showing specificity for each plant subspecies were found. Our study highlights the combined influences of bedrock and plant genotype on fungal recruitment into cushion soils and suggests the coexistence of two mechanisms, an indirect selection resulting from the colonization of an engineered soil by free-living saprobes and a direct selection resulting from direct plant-fungi interactions.


Upregulation of innate antiviral restricting factor expression in the cord blood and decidual tissue of HIV-infected mothers.
Programs for the prevention of mother-to-child transmission of HIV have reduced the transmission rate of perinatal HIV infection and have thereby increased the number of HIV-exposed uninfected (HEU) infants. Natural immunity to HIV-1 infection in both mothers and newborns needs to be further explored. In this study, we compared the expression of antiviral restricting factors in HIV-infected pregnant mothers treated with antiretroviral therapy (ART) in pregnancy (n=23) and in cord blood (CB) (n=16), placental tissues (n=10-13) and colostrum (n=5-6) samples and compared them to expression in samples from uninfected (UN) pregnant mothers (n=21). Mononuclear cells (MNCs) were prepared from maternal and CB samples following deliveries by cesarean section. Maternal (decidua) and fetal (chorionic villus) placental tissues were obtained, and colostrum was collected 24 h after delivery. The mRNA and protein expression levels of antiviral factors were then evaluated. We observed a significant increase in the mRNA expression levels of antiviral factors in MNCs from HIV-infected mothers and CB, including the apolipoprotein B mRNA-editing enzyme 3G (A3G), A3F, tripartite motif family-5α (TRIM-5α), TRIM-22, myxovirus resistance protein A (MxA), stimulator of interferon (IFN) genes (STING) and IFN-β, compared with the levels detected in uninfected (UN) mother-CB pairs. Moreover, A3G transcript and protein levels and α-defensin transcript levels were decreased in the decidua of HIV-infected mothers. Decreased TRIM-5α protein levels in the villi and increased STING mRNA expression in both placental tissues were also observed in HIV-infected mothers compared with uninfected (UN) mothers. Additionally, colostrum cells from infected mothers showed increased tetherin and IFN-β mRNA levels and CXCL9 protein levels. The data presented here indicate that antiviral restricting factor expression can be induced in utero in HIV-infected mothers. Future studies are warranted to determine whether this upregulation of antiviral factors during the perinatal period has a protective effect against HIV-1 infection.


The Influence of Supervisory Neglect on Subtypes of Emerging Adult Substance Use After Controlling for Familial Factors, Relationship Status, and Individual Traits.
This study is the first to explore how child supervisory neglect influences patterns of substance use among young adults. This study investigated patterns of substance use among males and females, 18 to 24 years old, after controlling for adolescent parental drinking, living with parents, relationship status, delinquency, and depression. The study sample (N=10,618) included individuals who participated in Waves I (1994-1995) and III (2001-2002) of the National Longitudinal Study of Adolescent Health (Add Health). The study used latent class analysis to ascertain how patterns of substance use emerged as distinct classes. For both males and females, we identified the following 4 classes of substance use: (1) heavy polysubstance use, (2) moderate polysubstance use, (3) alcohol and marijuana, and (4) low-use substance use patterns. Multinomial logistic regression indicated that, for both males and females 18 to 24 years old, experiencing supervisory neglect, being depressed, being single, and engaging in adolescent delinquency serve as risk factors for heavy polysubstance use class membership. Conversely, being black or Hispanic lowered the likelihood of polysubstance use for males and females. For females only, living with parents served as a protective factor that reduced the risk of membership in heavy polysubstance use, moderate polysubstance use, and alcohol and marijuana classes. For males only, being less educated increased the risk of heavy polysubstance use class membership. Results from this exploratory study underscore the enduring effect of supervisory neglect on substance use among male and female young adults. Future studies should explore whether these relationships hold over time.


The unfinished saga of invasive procedures for secondary mitral regurgitation.
Secondary mitral regurgitation (MR) is a common valvular heart disease. Its prognostic burden in patients suffering from idiopathic or ischemic cardiomyopathy (ICM) with left ventricular (LV) dysfunction/dilation has been clearly demonstrated. Severe secondary MR is associated with an increased mortality and frequent heart failure hospitalizations. Although guideline-directed medical therapy (GDMT) is the cornerstone of the management of secondary MR, a certain proportion of patients remain symptomatic. For these patients, several surgical techniques have been progressively developed during the last few decades (replacement, repair, sub-valvular apparatus interventions and other ventricular approaches). In the absence of evidence-based medicine, the benefits of these surgical procedures remains controversial, leading to a low level of recommendation in the guidelines. One way to anticipate the future is to look to the past. Recent prospective randomized trials evaluated surgical and percutaneous techniques and led to a better understanding of how best to treat this disease. In this article, we aim to describe the saga of the surgical and percutaneous treatments for secondary MR throughout the previous decades.


Cleavage C-terminal to Asp leads to covalent crosslinking of long-lived human proteins.
With age, long-lived proteins in the human body deteriorate, which can have consequences both for aging and disease. The aging process is often associated with the formation of covalently crosslinked proteins. Currently our knowledge of the mechanism of formation of these crosslinks is limited. In this study, proteomics was used to characterize sites of covalent protein-protein crosslinking and identify a novel mechanism of protein-protein crosslinking in the adult human lens. In this mechanism, Lys residues are crosslinked to C-terminal Asp residues that are formed by non-enzymatic protein truncation. Ten different crosslinks were identified in major lens proteins such as αA-crystallin, αB-crystallin and AQP0. Crosslinking in AQP0 increased significantly with age and also increased significantly in cataract lenses compared with normal lenses. Using model peptides, a mechanism of formation of the Lys-Asp crosslink was elucidated. The mechanism involves spontaneous peptide cleavage on the C-terminal side of Asp residues which can take place in the pH range 5-7.4. Cleavage appears to involve attack by the side chain carboxyl group on the adjacent peptide bond, resulting in the formation of a C-terminal Asp anhydride. This anhydride intermediate can then either react with water to form Asp, or with a nucleophile, such as a free amine group to form a crosslink. If an ε-amino group of Lys or an N-terminal amine group attacks the anhydride, a covalent protein-protein crosslink will be formed. This bi-phasic mechanism represents the first report to link two spontaneous events: protein cleavage and crosslinking that are characteristic of long-lived proteins.


Evaluation of yield of currently available diagnostics by sample type to optimize detection of respiratory pathogens in patients with a community-acquired pneumonia.
For the detection of respiratory pathogens, the sampling strategy may influence the diagnostic yield. Ideally, samples from the lower respiratory tract are collected, but they are difficult to obtain. In this study, we compared the diagnostic yield in sputum and oropharyngeal samples (OPS) for the detection of respiratory pathogens in patients with community-acquired pneumonia (CAP), with the objective to optimize our diagnostic testing algorithm. Matched sputum samples, OPS, blood cultures, serum, and urine samples were taken from patients (>18 years) with CAP and tested for the presence of possible respiratory pathogens using bacterial cultures, PCR for 17 viruses and five bacteria and urinary antigen testing. When using only conventional methods, that is, blood cultures, sputum culture, urinary antigen tests, a pathogen was detected in 49·6% of patients (n = 57). Adding molecular detection assays increased the yield to 80%. A pathogen was detected in 77 of the 115 patients in OPS or sputum samples by PCR. The sensitivity of the OPS was lower than that of the sputum samples (57% versus 74%). In particular, bacterial pathogens were more often detected in sputum samples. The sensitivity of OPS for the detection of most viruses was higher than in sputum samples (72% versus 66%), except for human rhinovirus and respiratory syncytial virus. Addition of PCR on both OPS and sputum samples significantly increased the diagnostic yield. For molecular detection of bacterial pathogens, a sputum sample is imperative, but for detection of most viral pathogens, an OPS is sufficient.


Migraine, stroke and patent foramen ovale: a dangerous trio?
The relationship between migraine, stroke and patent foramen ovale (PFO) has been the subject of considerable research efforts. Indeed, a lot of interest has focused on the potential benefits of percutaneous PFO closure. The aim of this article is to review data from the currently available literature. A total of 10 relevant studies were found in the literature, for a total of 1038 patients undergoing percutaneous PFO closure after events of cryptogenic stroke. Thirty-eight percent of these patients suffered from migraine. Combining the results of the available studies, 72% of patients were cured or improved significantly. Analysing the results according to migraine type, 81% of patients with migraine with aura had complete resolution or significantly improved as compared to 72% of patients with migraine without aura. Several limitations and drawbacks exist, however, and they are extensively discussed in this paper.