Population genetic structure and connectivity of the seagrass Thalassia hemprichii in the Western Indian Ocean is influenced by predominant ocean currents.
This study is the first large-scale genetic population study of a widespread climax species of seagrass, Thalassia hemprichii, in the Western Indian Ocean (WIO). The aim was to understand genetic population structure and connectivity of T. hemprichii in relation to hydrodynamic features. We genotyped 205 individual seagrass shoots from 11 sites across the WIO, spanning over a distance of ~2,700 km, with twelve microsatellite markers. Seagrass shoots were sampled in Kenya, Tanzania (mainland and Zanzibar), Mozambique, and Madagascar: 4-26°S and 33-48°E. We assessed clonality and visualized genetic diversity and genetic population differentiation. We used Bayesian clustering approaches (TESS) to trace spatial ancestry of populations and used directional migration rates (DivMigrate) to identify sources of gene flow. We identified four genetically differentiated groups: (a) samples from the Zanzibar channel; (b) Mozambique; (c) Madagascar; and (d) the east coast of Zanzibar and Kenya. Significant pairwise population genetic differentiation was found among many sites. Isolation by distance was detected for the estimated magnitude of divergence (D EST), but the three predominant ocean current systems (i.e., East African Coastal Current, North East Madagascar Current, and the South Equatorial Current) also determine genetic connectivity and genetic structure. Directional migration rates indicate that Madagascar acts as an important source population. Overall, clonality was moderate to high with large differences among sampling sites, indicating relatively low, but spatially variable sexual reproduction rates. The strongest genetic break was identified for three sites in the Zanzibar channel. Although isolation by distance is present, this study suggests that the three regionally predominant ocean current systems (i.e., East African Coastal Current, North East Madagascar Current, and the South Equatorial Current) rather than distance determine genetic connectivity and structure of T. hemprichii in the WIO. If the goal is to maintain genetic connectivity of T. hemprichii within the WIO, conservation planning and implementation of marine protection should be considered at the regional scale-across national borders.


A comparison of the strength of biodiversity effects across multiple functions.
In order to predict which ecosystem functions are most at risk from biodiversity loss, meta-analyses have generalised results from biodiversity experiments over different sites and ecosystem types. In contrast, comparing the strength of biodiversity effects across a large number of ecosystem processes measured in a single experiment permits more direct comparisons. Here, we present an analysis of 418 separate measures of 38 ecosystem processes. Overall, 45 % of processes were significantly affected by plant species richness, suggesting that, while diversity affects a large number of processes not all respond to biodiversity. We therefore compared the strength of plant diversity effects between different categories of ecosystem processes, grouping processes according to the year of measurement, their biogeochemical cycle, trophic level and compartment (above- or belowground) and according to whether they were measures of biodiversity or other ecosystem processes, biotic or abiotic and static or dynamic. Overall, and for several individual processes, we found that biodiversity effects became stronger over time. Measures of the carbon cycle were also affected more strongly by plant species richness than were the measures associated with the nitrogen cycle. Further, we found greater plant species richness effects on measures of biodiversity than on other processes. The differential effects of plant diversity on the various types of ecosystem processes indicate that future research and political effort should shift from a general debate about whether biodiversity loss impairs ecosystem functions to focussing on the specific functions of interest and ways to preserve them individually or in combination.


Reactivation of Dihydroorotate Dehydrogenase-Driven Pyrimidine Biosynthesis Restores Tumor Growth of Respiration-Deficient Cancer Cells.
Cancer cells without mitochondrial DNA (mtDNA) do not form tumors unless they reconstitute oxidative phosphorylation (OXPHOS) by mitochondria acquired from host stroma. To understand why functional respiration is crucial for tumorigenesis, we used time-resolved analysis of tumor formation by mtDNA-depleted cells and genetic manipulations of OXPHOS. We show that pyrimidine biosynthesis dependent on respiration-linked dihydroorotate dehydrogenase (DHODH) is required to overcome cell-cycle arrest, while mitochondrial ATP generation is dispensable for tumorigenesis. Latent DHODH in mtDNA-deficient cells is fully activated with restoration of complex III/IV activity and coenzyme Q redox-cycling after mitochondrial transfer, or by introduction of an alternative oxidase. Further, deletion of DHODH interferes with tumor formation in cells with fully functional OXPHOS, while disruption of mitochondrial ATP synthase has little effect. Our results show that DHODH-driven pyrimidine biosynthesis is an essential pathway linking respiration to tumorigenesis, pointing to inhibitors of DHODH as potential anti-cancer agents.


Unusual binding of a potential biomarker with human serum albumin.
This study investigates the specific binding of a potential biomarker, [2,2'-bipyridyl]-3,3'-diol (BP(OH)2), with human serum albumin (HSA). The binding of BP(OH)2 at the two primary drug-binding sites on HSA (Sudlow's sites I and II) is explored by a competitive-binding study and monitored by considering the green-light emission from its diketo tautomer. Warfarin is used as a marker for site I and dansyl-L-proline (DP) as a competitor for site II. Steady-state and time-resolved fluorescence measurements affirm that neither of Sudlow's sites is the binding locus of BP(OH)2. To gain an idea regarding the probable binding site of BP(OH)2, we perform molecular-docking studies, which reveal a close proximity of the probe to Trp-214 in subdomain IIA of HSA. Confirmation of this contention is achieved by studying the quenching of the fluorescence of Trp-214 in the presence of BP(OH)2. Moreover, static quenching seems to be responsible for the depletion of the fluorescence of Trp-214, as manifested by the invariance of the intrinsic fluorescence lifetime of Trp-214, as a function of the concentration of BP(OH)2. Based on displacement and quenching studies, supported by molecular docking, we propose that BP(OH)2 binds in a cleft that separates subdomains IIIA and IIB, which is in close proximity to Trp-214.


Is FMR1 CGG repeat length a predictor of in vitro fertilization stimulation response or outcome?
To study a broad range of FMR1 CGG repeat lengths and assisted reproduction technology (ART) outcomes. Retrospective cohort study. Private ART practice. Fresh autologous ART stimulation cycles. None. Oocyte yield, live birth. We screened 14,088 fresh autologous ART cycles from 2012 to 2015, of which 4,690 cycles in 3,290 patients met the inclusion criteria. The FMR1 repeat length was statistically significantly but weakly associated with oocyte yield and other markers of ovarian response. The receiver operating characteristic curve analysis suggested extremely limited predictive ability. Moreover, the FMR1 repeat length was not statistically significantly associated with outcomes in multivariable models, including other markers of ovarian reserve. The FMR1 repeat length was not associated with embryo quality or live birth. Only patient age had a strong ability to predict live birth. The FMR1 repeat length is associated with ART response, but only weakly. It provides no incremental predictive ability beyond the conventionally used predictors, including patient age, antimüllerian hormone concentration, antral follicle count, and follicle-stimulating hormone level. These data suggest a possible role of the FMR1 repeat length within the normal range in ovarian response but demonstrate no clinically relevant indication for testing FMR1 as a predictor of ART outcomes.


Progression characteristics of ellipsoid zone loss in macular telangiectasia type 2.
To investigate the progression characteristics of ellipsoid zone (EZ) loss in eyes with macular telangiectasia type 2 (MacTel) as reflected by area and linear measurements, and their relevance for visual acuity. Participants were selected from the MacTel Study cohort. Linear and area measurements of EZ loss were performed in Spectral-Domain Optical Coherence Tomograph (SD-OCT) volume scans. Progression characteristics and correlations between linear and area measurements were analysed using linear mixed effects models. A total of 134 eyes of 70 patients were included (85 eyes with follow-up, mean 4.7 years, range: 1.4-8 years). Ellipsoid zone (EZ) loss significantly progressed at a mean annual increment of 0.057 mm2 (p = 0.005). The progression rate was non-linear and interacted significantly with initial EZ lesion size indicating an exponential growth before reaching a plateau. There was a strong heterogeneity in area sizes between fellow eyes. EZ break length had a significant linear effect on EZ break area (b = 1.06, p < 0.001) and could predict it. The location of the EZ break had a significant impact on visual acuity. Ellipsoid zone (EZ) loss in MacTel has a non-linear progression characteristic, and its rate depends on area size at baseline, which must be taken into account at sample selection in clinical trials. Our results show a good correlation of linear and area measures of EZ loss and a segregation of best-corrected visual acuity by EZ location, which may help routine clinical practice.


Metacognition: ideas and insights from neuro- and educational sciences.
Metacognition comprises both the ability to be aware of one's cognitive processes (metacognitive knowledge) and to regulate them (metacognitive control). Research in educational sciences has amassed a large body of evidence on the importance of metacognition in learning and academic achievement. More recently, metacognition has been studied from experimental and cognitive neuroscience perspectives. This research has started to identify brain regions that encode metacognitive processes. However, the educational and neuroscience disciplines have largely developed separately with little exchange and communication. In this article, we review the literature on metacognition in educational and cognitive neuroscience and identify entry points for synthesis. We argue that to improve our understanding of metacognition, future research needs to (i) investigate the degree to which different protocols relate to the similar or different metacognitive constructs and processes, (ii) implement experiments to identify neural substrates necessary for metacognition based on protocols used in educational sciences, (iii) study the effects of training metacognitive knowledge in the brain, and (iv) perform developmental research in the metacognitive brain and compare it with the existing developmental literature from educational sciences regarding the domain-generality of metacognition.


Chemical and ecotoxicological characterization of ashes obtained from sewage sludge combustion in a fluidised-bed reactor.
In 1999, the DEECA/INETI and the UBiA/FCT/UNL started a researching project on the partition of heavy metals during the combustion of stabilised sewage sludge (Biogran), in a fluidised-bed reactor, and on the quality of the bottom ashes and fly ashes produced. This project was entitled Bimetal and was funded by the Portuguese Foundation for Science and Technology. In this paper only the results on the combustion of Biogran are reported. The combustion process was performed in two different trials, in which different amounts of sewage sludge and time of combustion were applied. Several ash samples were collected from the bed (bottom ashes) and from two cyclones (first cyclone and second cyclone ashes). Sewage sludge, bed material (sand) and ash samples were submitted to the leaching process defined in the European leaching standard EN 12457-2. The eluates were characterized for a set of inorganic chemical species. The ecotoxicological levels of the eluates were determined for two biological indicators (Vibrio fischeri and Daphnia magna). The results were compared with the limit values of the CEMWE French Regulation. The samples were also ranked according to an index based on the chemical characterization of the eluates. It was observed an increase of the concentration of metals along the combustion system. The ashes trapped in the second cyclone, for both combustion trials, showed the highest concentration of metals in the eluates. Chemically, the ashes of the second cyclone were the most different ones. In the ecotoxicological point of view, the ecotoxicity levels of the eluates of the ashes, for both combustion cycles, did not follow the same pattern as observed for the chemical characterization. The ashes of the first cyclone showed the highest ecotoxicity levels for V. fischeri and D. magna. This difference on chemical and ecotoxicological results proves the need for performing both chemical and ecotoxicological characterizations of the sub-products of such type of thermal processes.


Foods and Drinks Available from Urban Food Pantries: Nutritional Quality by Item Type, Sourcing, and Distribution Method.
The overall nutritional quality of foods/drinks available at urban food pantries is not well established. In a study of 50 pantries listed as operating in the Bronx, NY, data on food/drink type (fresh, shelf-stable, refrigerated/frozen) came from direct observation. Data on food/drink sourcing (food bank or other) and distribution (prefilled bag vs. client choice for a given client's position in line) came from semi-structured interviews with pantry workers. Overall nutritional quality was determined using NuVal® scores (range 1-100; higher score indicates higher nutritional quality). Twenty-nine pantries offered zero nutrition at listed times (actually being closed or having no food/drinks in stock). Of the 21 pantries that were open as listed and had foods/drinks to offer, 12 distributed items in prefilled bags (traditional pantries), 9 allowed for client choice. Mean NuVal® scores were higher for foods/drinks available from client-choice pantries than traditional pantries (69.3 vs. 57.4), driven mostly by sourcing fresh items (at 28.3% of client-choice pantries vs. 4.8% of traditional pantries). For a hypothetical 'balanced basket' of one of each fruit, vegetable, grain, dairy and protein item, highest-NuVal® items had a mean score of 98.8 across client-choice pantries versus 96.6 across traditional pantries; lowest-NuVal® items had mean scores of 16.4 and 35.4 respectively. Pantry workers reported lower-scoring items (e.g., white rice) were more popular-appeared in early bags or were selected first-leaving higher-scoring items (e.g., brown rice) for clients later in line. Fewer than 50% of sampled pantries were open and had food/drink to offer at listed times. Nutritional quality varied by item type and sourcing and could also vary by distribution method and client position in line. Findings suggest opportunities for pantry operation, client and staff education, and additional research.


A synaptic nidogen: developmental regulation and role of nidogen-2 at the neuromuscular junction.
The skeletal neuromuscular junction is a useful model for elucidating mechanisms that regulate synaptogenesis. Developmentally important intercellular interactions at the neuromuscular junction are mediated by the synaptic portion of a basal lamina that completely ensheaths each muscle fiber. Basal laminas in general are composed of four main types of glycosylated proteins: laminins, collagens IV, heparan sulfate proteoglycans and nidogens (entactins). The portion of the muscle fiber basal lamina that passes between the motor nerve terminal and postsynaptic membrane has been shown to bear distinct isoforms of the first three of these. For laminins and collagens IV, the proteins are deposited by the muscle; a synaptic proteoglycan, z-agrin, is deposited by the nerve. In each case, the synaptic isoform plays key roles in organizing the neuromuscular junction. Here, we analyze the fourth family, composed of nidogen-1 and -2. In adult muscle, nidogen-1 is present throughout muscle fiber basal lamina, while nidogen-2 is concentrated at synapses. Nidogen-2 is initially present throughout muscle basal lamina, but is lost from extrasynaptic regions during the first three postnatal weeks. Neuromuscular junctions in mutant mice lacking nidogen-2 appear normal at birth, but become topologically abnormal as they mature. Synaptic laminins, collagens IV and heparan sulfate proteoglycans persist in the absence of nidogen-2, suggesting the phenotype is not secondary to a general defect in the integrity of synaptic basal lamina. Further genetic studies suggest that synaptic localization of each of the four families of synaptic basal lamina components is independent of the other three. All four core components of the basal lamina have synaptically enriched isoforms. Together, they form a highly specialized synaptic cleft material. Individually, they play distinct roles in the formation, maturation and maintenance of the neuromuscular junction.


Hybrid therapy as first-line regimen for Helicobacter pylori eradication in a high clarithromycin resistance area: a prospective open-label trial.
Hybrid therapy is a promising first-line regimen for Helicobacter pylori (H. pylori) eradication. We evaluated a hybrid therapy, assessing the impact of antibiotic resistance on eradication outcome. This was a prospective study that included 155 treatment-naïve patients diagnosed with H. pylori infection by positive CLO-test, confirmed with histology and/or culture. The hybrid therapy consisted of 40 mg esomeprazole and 1 g amoxicillin for 14 days, with the addition of 500 mg clarithromycin and 500 mg metronidazole for the final 7 days (all b.i.d.). Eradication was defined by negative 13C-urea breath test or histology. The eradication rates were 85.8% (133/155; 95% confidence interval [CI] 79.4-90.5%) by intention-to-treat and 90.2% (129/143; 95%CI 84.1-94.2%) by per-protocol analysis in a setting of high antibiotic resistance (clarithromycin 25.9%, metronidazole 31.1%, dual resistance 8.9%). Adverse events occurred in 29.7% and 1.3% discontinued treatment because of adverse events. Adherence >90% was achieved in 96.6%. The eradication rate in patients with dual clarithromycin/metronidazole resistance (50%) was markedly lower compared to those with single clarithromycin resistance (91.4%), single metronidazole resistance (90.5%) or dual susceptibility (97.8%). Dual resistance was the only factor to correlate with the failure of hybrid therapy (odds ratio 14.4, 95%CI 3.8-54.9, P=0.0003). Hybrid therapy is an effective and safe first-line regimen in populations with relatively high rates of antibiotic resistance. However, dual clarithromycin/metronidazole resistance may significantly compromise its efficacy.


Crystal growth inside an octant.
We study crystal growth inside an infinite octant on a cubic lattice. The growth proceeds through the deposition of elementary cubes into inner corners. After rescaling by the characteristic size, the interface becomes progressively more deterministic in the long-time limit. Utilizing known results for the crystal growth inside a two-dimensional corner, we propose a hyperbolic partial differential equation for the evolution of the limiting shape. This equation is interpreted as a Hamilton-Jacobi equation, which helps in finding an analytical solution. Simulations of the growth process are in excellent agreement with analytical predictions. We then study the evolution of the subleading correction to the volume of the crystal, the asymptotic growth of the variance of the volume of the crystal, and the total number of inner and outer corners. We also show how to generalize the results to arbitrary spatial dimension.


Dissecting the Molecular Structure of the Air/Water Interface from Quantum Simulations of the Sum-Frequency Generation Spectrum.
The molecular characterization of the air/water interface is a key step in understanding fundamental multiphase phenomena ranging from heterogeneous chemical processes in the atmosphere to the hydration of biomolecules. The apparent simplicity of the air/water interface, however, masks an underlying complexity associated with the dynamic nature of the water hydrogen-bond network that has so far hindered an unambiguous characterization of its microscopic properties. Here, we demonstrate that the application of quantum many-body molecular dynamics, which enables spectroscopically accurate simulations of water from the gas to the condensed phase, leads to a definitive molecular-level picture of the interface region. For the first time, excellent agreement is obtained between the simulated vibrational sum-frequency generation spectrum and the most recent state-of-the-art measurements, without requiring any empirical frequency shift or ad hoc scaling of the spectral intensity. A systematic dissection of the spectral features demonstrates that a rigorous representation of nuclear quantum effects as well as of many-body energy and electrostatic contributions is necessary for a quantitative reproduction of the experimental data. The unprecedented accuracy of the simulations presented here indicates that quantum many-body molecular dynamics can enable predictive studies of aqueous interfaces, which by complementing analogous experimental measurements will provide unique molecular insights into multiphase and heterogeneous processes of relevance in chemistry, biology, materials science, and environmental research.


The Haverstraw experience: the first tobacco product display ban in the United States.
In April 2012, the village of Haverstraw, New York, passed the first tobacco retail display ban in the United States. Community groups funded by the New York State Department of Health Tobacco Control Program mobilized community members to support an initiative to protect youths in their area from tobacco marketing via methods consistent with a community transformation framework. The law was soon rescinded after 7 tobacco companies and the New York Association of Convenience Stores filed a federal lawsuit against the village that challenged the law's constitutionality. We discuss lessons learned and next steps for adoption of local point-of-sale policies.


Atmospheric seasonal forecasts of the twentieth century: multi-decadal variability in predictive skill of the winter North Atlantic Oscillation (NAO) and their potential value for extreme event attribution.
Based on skill estimates from hindcasts made over the last couple of decades, recent studies have suggested that considerable success has been achieved in forecasting winter climate anomalies over the Euro-Atlantic area using current-generation dynamical forecast models. However, previous-generation models had shown that forecasts of winter climate anomalies in the 1960s and 1970s were less successful than forecasts of the 1980s and 1990s. Given that the more recent decades have been dominated by the North Atlantic Oscillation (NAO) in its positive phase, it is important to know whether the performance of current models would be similarly skilful when tested over periods of a predominantly negative NAO. To this end, a new ensemble of atmospheric seasonal hindcasts covering the period 1900-2009 has been created, providing a unique tool to explore many aspects of atmospheric seasonal climate prediction. In this study we focus on two of these: multi-decadal variability in predicting the winter NAO, and the potential value of the long seasonal hindcast datasets for the emerging science of probabilistic event attribution. The existence of relatively low skill levels during the period 1950s-1970s has been confirmed in the new dataset. The skill of the NAO forecasts is larger, however, in earlier and later periods. Whilst these inter-decadal differences in skill are, by themselves, only marginally statistically significant, the variations in skill strongly co-vary with statistics of the general circulation itself suggesting that such differences are indeed physically based. The mid-century period of low forecast skill coincides with a negative NAO phase but the relationship between the NAO phase/amplitude and forecast skill is more complex than linear. Finally, we show how seasonal forecast reliability can be of importance for increasing confidence in statements of causes of extreme weather and climate events, including effects of anthropogenic climate change.


The (unusual) heuristic value of Hox gene clusters; a matter of time?
Ever since their first report in 1984, Antennapedia-type homeobox (Hox) genes have been involved in such a series of interesting observations, in particular due to their conserved clustered organization between vertebrates and arthropods, that one may legitimately wonder about the origin of this heuristic value. In this essay, I first consider different examples where Hox gene clusters have been instrumental in providing conceptual advances, taken from various fields of research and mostly involving vertebrate embryos. These examples touch upon our understanding of genomic evolution, the revisiting of 19th century views on the relationships between development and evolution and the building of a new framework to understand long-range and pleiotropic gene regulation during development. I then discuss whether the high value of the Hox gene family, when considered as an epistemic object, is related to its clustered structure (and the absence thereof in some animal species) and, if so, what is it in such particular genetic oddities that made them so generous in providing the scientific community with interesting information.


Comparison of the timed 25-foot and the 100-meter walk as performance measures in multiple sclerosis.
Ambulation impairment is a major component of physical disability in multiple sclerosis (MS) and a major target of rehabilitation programs. Outcome measures commonly used to evaluate walking capacities suffer from several limitations. To define and validate a new test that would overcome the limitations of current gait evaluations in MS and ultimately better correlate with the maximum walking distance (MWD). The authors developed the Timed 100-Meter Walk Test (T100MW), which was compared with the Timed 25-Foot Walk Test (T25FW). For the T100MW, the subject is invited to walk 100 m as fast as he/she can. In MS patients and healthy control volunteers, the authors measured the test-retest and interrater intraclass correlation coefficient. Spearman rank correlations were obtained between the T25FW, the T100MW, the Expanded Disability Status Scale (EDSS), and the MWD. The coefficient of variation, Bland-Altman plots, the coefficient of determination, and the area under the receiver operator characteristic curve were measured. The mean walking speed (MWS) was compared between the 2 tests. A total of 141 MS patients and 104 healthy control volunteers were assessed. Minor differences favoring the T100MW over the T25FW were observed. Interestingly, the authors demonstrated a paradoxically higher MWS on a long (T100MW) rather than on a short distance walk test (T25FW). The T25FW and T100MW displayed subtle differences of reproducibility, variability, and correlation with MWD favoring the T100MW. The maximum walking speed of MS patients may be poorly estimated by the T25FW since MS patients were shown to walk faster over a longer distance.


The price of your soul: neural evidence for the non-utilitarian representation of sacred values.
Sacred values, such as those associated with religious or ethnic identity, underlie many important individual and group decisions in life, and individuals typically resist attempts to trade off their sacred values in exchange for material benefits. Deontological theory suggests that sacred values are processed based on rights and wrongs irrespective of outcomes, while utilitarian theory suggests that they are processed based on costs and benefits of potential outcomes, but which mode of processing an individual naturally uses is unknown. The study of decisions over sacred values is difficult because outcomes cannot typically be realized in a laboratory, and hence little is known about the neural representation and processing of sacred values. We used an experimental paradigm that used integrity as a proxy for sacredness and which paid real money to induce individuals to sell their personal values. Using functional magnetic resonance imaging (fMRI), we found that values that people refused to sell (sacred values) were associated with increased activity in the left temporoparietal junction and ventrolateral prefrontal cortex, regions previously associated with semantic rule retrieval. This suggests that sacred values affect behaviour through the retrieval and processing of deontic rules and not through a utilitarian evaluation of costs and benefits.


Prenatal screening for trisomy 18 with free beta human chorionic gonadotrophin as a marker.
To determine the relation between maternal serum alpha fetoprotein and free beta human chorionic gonadotrophin concentrations in pregnancies complicated by trisomy 18 and establish whether prenatal biochemical screening for this condition could be developed in a way similar to that proposed for trisomy 21. Serum alpha fetoprotein and free beta human chorionic gonadotrophin concentrations in women with singleton pregnancies affected by cytogenetically confirmed trisomy 18, uncomplicated by neural tube defect or ventral wall defect, were identified from prospective trisomy 21 screening programmes. Additionally, stored maternal serum from similar pregnancies was analysed retrospectively. Analyte concentrations from singleton unaffected pregnancies were identified from a prospective screening programme as controls. Statistical parameters of the affected and unaffected populations were compiled. Biochemical screening laboratories in Britain and the United States. 52 women with singleton pregnancies complicated by trisomy 18; control population of 6661 women with unaffected singleton pregnancies. Median values of each analyte and their distribution in the affected and unaffected populations; detection rate of trisomy 18 and the false positive rate. Maternal serum alpha fetoprotein and free beta human chorionic gonadotrophin concentrations were significantly lower in pregnancies complicated by trisomy 18 (median values 0.71 and 0.37 respectively). By using a multivariate risk algorithm incorporating maternal age risk of trisomy 18 and the concentration of the two biochemical markers it was predicted that 50% of trisomy 18 cases (unaffected by neural tube defect or ventral wall defect) could be detected with a 1% false positive rate. Second trimester biochemical screening for trisomy 18 could be a valuable addition to trisomy 21 screening programmes.


Paternal Genome Elimination in Liposcelis Booklice (Insecta: Psocodea).
How sex is determined in insects is diverse and dynamic, and includes male heterogamety, female heterogamety, and haplodiploidy. In many insect lineages, sex determination is either completely unknown or poorly studied. We studied sex determination in Psocodea-a species-rich order of insects that includes parasitic lice, barklice, and booklice. We focus on a recently discovered species of Liposcelis booklice (Psocodea: Troctomorpha), which are among the closest free-living relatives of parasitic lice. Using genetic, genomic, and immunohistochemical approaches, we show that this group exhibits paternal genome elimination (PGE), an unusual mode of sex determination that involves genomic imprinting. Controlled crosses, following a genetic marker over multiple generations, demonstrated that males only transmit to offspring genes they inherited from their mother. Immunofluorescence microscopy revealed densely packed chromocenters associated with H3K9me3-a conserved marker for heterochromatin-in males, but not in females, suggesting silencing of chromosomes in males. Genome assembly and comparison of read coverage in male and female libraries showed no evidence for differentiated sex chromosomes. We also found that females produce more sons early in life, consistent with facultative sex allocation. It is likely that PGE is widespread in Psocodea, including human lice. This order represents a promising model for studying this enigmatic mode of sex determination.


Computer-Assisted Corrective Osteotomy of Malunited Pediatric Radial Neck Fractures-Three-Dimensional Postoperative Accuracy and Clinical Outcome.
Neglected or incorrect treatment of pediatric radial neck fractures may lead to symptomatic malunions. Computer-assisted corrective osteotomies with patient-specific guides have been proposed as a promising technique for the reconstruction of malunited long bone deformities. The aim of this study was to evaluate the accuracy and clinical outcome of this technique in children with malunited fractures of the radial neck. Four children [2 boys, 2 girls; mean age 12 (10-16) years] underwent computer-assisted closing wedge osteotomy of the radial neck. The contralateral uninjured side was used as a reconstruction template. Computed tomography were performed 8 weeks postoperatively to confirm bony consolidation and to quantify residual 3D rotational and translational displacement error. Clinical outcome [pain, range of motion (ROM)] and overall satisfaction were documented. Preoperative subluxation of the radial head could be corrected in 2 of 3 patients. One patient had to be revised because of secondary traumatic loss of reduction. At the last follow-up [mean 16 (range, 12-24) months], all patients were pain free for activities of daily living (preoperative pain: visual analog scale 6). Pain during sport activities could be substantially reduced (visual analog scale 8→2). Although the procedure failed to improve ROM, none of the patients had limitations regarding work, daily, or sports activities. Yet, restricted ROM was considered as a cosmetic problem in 1 patient. Full consolidation of the osteotomy site, with no signs of avascular necrosis of the radial head, was achieved in all patients. The deformity could be substantially reduced, from a 3D angle of 13-40 degrees to 3-7 degrees (58%-89% deformity correction). Computer-assisted corrective osteotomy is a novel technique for the treatment of radial neck malunions that led to adequate pain reduction and 3D accuracy of deformity correction in our small case series. Despite the lack of improved ROM, all patients were satisfied and would undergo the same procedure again.


High-level systemic expression of conserved influenza epitope in plants on the surface of rod-shaped chimeric particles.
Recombinant viruses based on the cDNA copy of the tobacco mosaic virus (TMV) genome carrying different versions of the conserved M2e epitope from influenza virus A cloned into the coat protein (CP) gene were obtained and partially characterized by our group previously; cysteines in the human consensus M2e sequence were changed to serine residues. This work intends to show some biological properties of these viruses following plant infections. Agroinfiltration experiments on Nicotiana benthamiana confirmed the efficient systemic expression of M2e peptides, and two point amino acid substitutions in recombinant CPs significantly influenced the symptoms and development of viral infections. Joint expression of RNA interference suppressor protein p19 from tomato bushy stunt virus (TBSV) did not affect the accumulation of CP-M2e-ser recombinant protein in non-inoculated leaves. RT-PCR analysis of RNA isolated from either infected leaves or purified TMV-M2e particles proved the genetic stability of TMV‑based viral vectors. Immunoelectron microscopy of crude plant extracts demonstrated that foreign epitopes are located on the surface of chimeric virions. The rod‑shaped geometry of plant-produced M2e epitopes is different from the icosahedral or helical filamentous arrangement of M2e antigens on the carrier virus-like particles (VLP) described earlier. Thereby, we created a simple and efficient system that employs agrobacteria and plant viral vectors in order to produce a candidate broad-spectrum flu vaccine.


Tooth size variation related to age in Amboseli baboons.
We measured the molar size from a single population of wild baboons from Amboseli (Kenya), both females (n=57) and males (n=50). All the females were of known age; the males represented a mix of known-age individuals (n=31) and individuals with ages estimated to within 2 years (n=19). The results showed a significant reduction in the mesiodistal length of teeth in both sexes as a function of age. Overall patterns of age-related change in tooth size did not change whether we included or excluded the individuals of estimated age, but patterns of statistical significance changed as a result of changed sample sizes. Our results demonstrate that tooth length is directly related to age due to interproximal wearing caused by M2 and M3 compression loads. Dental studies in primates, including both fossil and extant species, are mostly based on specimens obtained from osteological collections of varying origins, for which the age at death of each individual in the sample is not known. Researchers should take into account the phenomenon of interproximal attrition leading to reduced tooth size when measuring tooth length for ondontometric purposes.


Folic Acid Supplementation of Female Mice, with or without Vitamin B-12, before and during Pregnancy and Lactation Programs Adiposity and Vascular Health in Adult Male Offspring.
The developmental origins of health and disease theory suggest that disturbances in the fetal and early postnatal environment contribute to chronic adulthood diseases, such as type 2 diabetes and cardiovascular disease. Greater adiposity and insulin resistance have been reported in children of women with high erythrocyte folate but poor vitamin B-12 status during pregnancy. The mechanisms underlying this relation are not known. The objective of this study was to investigate the effects of maternal supplemental folic acid, with or without vitamin B-12, on adiposity, glucose homeostasis, and vascular health in adult male offspring mice. Female C57BL/6J mice were fed a control diet (M-CON, 2 mg folic acid/kg, 50 μg vitamin B-12/kg) or a folic acid-supplemented diet with [10 mg folic acid/kg, 50 μg vitamin B-12/kg (SFA+B12)] or without [10 mg folic acid/kg, no vitamin B-12 (SFA-B12)] vitamin B-12 for 6 wk before mating and during pregnancy and lactation. The offspring were weaned onto a control diet (16% energy from fat) or a western diet (45% energy from fat) until 23 wk of age. The effects of maternal diet on adiposity, vascular function, and glucose tolerance were assessed in 6 groups of adult male offspring: control diet-fed M-CON, SFA+B12, and SFA-B12 and western diet-fed M-CON, SFA+B12, and SFA-B12. Control and western diet-fed SFA-B12 and SFA+B12 offspring had smaller visceral and subcutaneous adipose tissue than M-CON offspring (P < 0.05). Control SFA-B12 and SFA+B12 offspring had lower serum total adiponectin and vitamin B-12 concentrations and lower NADPH oxidase 2 expression in aorta compared with M-CON offspring (P < 0.05). These effects were not observed in western diet-fed offspring. Folic acid supplementation of female mice before and during pregnancy and lactation, with or without dietary vitamin B-12, affects adult male offspring adiposity, vascular function, and one-carbon metabolism in those fed a control diet but not a western diet.


The impact of preoperative stoma site marking on the incidence of complications, quality of life, and patient's independence.
Preoperative stoma site marking and counseling aim to improve patients' rehabilitation and adaptation to a new medical condition. Objective studies are needed to provide evidence of the impact of care by stoma therapists. Key quality indicators include patients' quality of life, independence, and complication rates as affected by the variable modes of stoma site marking and planning. The aim of this study was to evaluate the impact of preoperative stoma site marking on patients' quality of life, independence, and complication rates. : A validated stoma quality-of-life questionnaire was used as the main assessment tool. Complications were noted on regular postoperative visits. This is a single-center, clinical study. The study was conducted at the Rambam Health Care Campus in Haifa, Israel. Rambam Health Care Campus is a tertiary university hospital. All patients who underwent an elective stoma creation between 2006 and 2008 were included. Evaluated parameters included demographics, stoma type, marking status, complication rates, quality of life, and independence parameters. : One hundred five patients (60 men and 45 women) were included, of whom 52 (49.5%) were preoperatively marked and 53 (50.5%) were not marked. Sixty stomas (57%) were permanent, and 45 (43%) stomas were temporary. The quality of life of patients whose stoma sites were preoperatively marked was significantly better than that of the unmarked patients (p < 0.05 in 18 of 20 items), their independence parameters were significantly better, and their complication rates significantly lower. All of these results were significant irrespective of the stoma type. Preoperative stoma site marking is crucial for improving patients' postoperative quality of life, promoting their independence, and reducing the rates of postoperative complications. The role of the enterostomal therapist is very important in the ostomates' pre- and postoperative care.


Nutrition intervention trials in Linxian, China: supplementation with specific vitamin/mineral combinations, cancer incidence, and disease-specific mortality in the general population.
Epidemiologic evidence indicates that diets high in fruits and vegetables are associated with a reduced risk of several cancers, including cancers of the esophagus and stomach. Vitamins and minerals in these foods may contribute to the reduced cancer risk. The people of Linxian County, China, have one of the world's highest rates of esophageal/gastric cardia cancer and a persistently low intake of several micronutrients. We sought to determine if dietary supplementation with specific vitamins and minerals can lower mortality from or incidence of cancer as well as mortality from other diseases in Linxian. Individuals of ages 40-69 were recruited in 1985 from four Linxian communes. Mortality and cancer incidence during March 1986-May 1991 were ascertained for 29,584 adults who received daily vitamin and mineral supplementation throughout this period. The subjects were randomly assigned to intervention groups according to a one-half replicate of a 2(4) factorial experimental design. This design enabled testing for the effects of four combinations of nutrients: (A) retinol and zinc; (B) riboflavin and niacin; (C) vitamin C and molybdenum; and (D) beta carotene, vitamin E, and selenium. Doses ranged from one to two times U.S. Recommended Daily Allowances. A total of 2127 deaths occurred among trial participants during the intervention period. Cancer was the leading cause of death, with 32% of all deaths due to esophageal or stomach cancer, followed by cerebrovascular disease (25%). Significantly (P = .03) lower total mortality (relative risk [RR] = 0.91; 95% confidence interval [CI] = 0.84-0.99) occurred among those receiving supplementation with beta carotene, vitamin E, and selenium. The reduction was mainly due to lower cancer rates (RR = 0.87; 95% CI = 0.75-1.00), especially stomach cancer (RR = 0.79; 95% CI = 0.64-0.99), with the reduced risk beginning to arise about 1-2 years after the start of supplementation with these vitamins and minerals. No significant effects on mortality rates from all causes were found for supplementation with retinol and zinc, riboflavin and niacin, or vitamin C and molybdenum. Patterns of cancer incidence, on the basis of 1298 cases, generally resembled those for cancer mortality. The findings indicate that vitamin and mineral supplementation of the diet of Linxian adults, particularly with the combination of beta carotene, vitamin E, and selenium, may effect a reduction in cancer risk in this population. The results on their own are not definitive, but the promising findings should stimulate further research to clarify the potential benefits of micronutrient supplements.


Genetic and environmental contributions to allergen sensitization in a Chinese twin study.
Allergic disease is on the rise worldwide. Effective prevention of allergic disease requires comprehensive understanding of the factors that contribute to its intermediate phenotypes, such as sensitization to common allergens. To estimate the degree of genetic and environmental contributions to sensitization to food and aeroallergens. Sensitization was defined as a positive skin prick test to an allergen. We calculated the zygosity-specific concordance rates and odds ratios (ORs) for sensitization to food and aeroallergens in 826 Chinese twin pairs [472 monozygotic (MZ) and 354 dizygotic (DZ)] aged 12-28 years. We also applied structural equation modelling procedures to estimate genetic and environmental influences on sensitization. The concordance rates and risk of sensitization in one twin given the presence vs. the absence of sensitization in the other twin were higher in MZ twins than those in DZ twins. However, a large number of MZ twins were discordant in sensitization to common allergens. These observations suggest both genetic and environmental factors influence sensitization. Consistently, the estimated heritability and individual environmental components of the liability to sensitization ranged from 0.51 to 0.68 and 0.32 to 0.49, respectively, based on the best-fitted structural equation model. We also observed high phenotypic correlations between sensitization to two aeroallergens (cockroach and dust mite: 0.83) and two food allergens (peanut and shellfish: 0.58), but only moderate correlations for the pairs between sensitization to a food and an aeroallergen (0.31-0.46). The shared genetic and environmental factors between paired sensitizations contribute to the observed correlations. We demonstrated that sensitization to common food and aeroallergens were influenced by both genetic and environmental factors. Moreover, we found that paired allergen sensitizations might share some common sets of genes and environmental factors. This study underscores the need to further delineate unique and/or pleiotropic genetic and environmental factors for allergen sensitization.


Coronary artery bypass grafting in hemodialysis-dependent patients: analysis of Japan Adult Cardiovascular Surgery Database.
Perioperative risk during coronary artery bypass grafting (CABG) is reportedly high in patients with chronic renal disease. We aimed to determine postoperative mortality and morbidity and identify the perioperative risk factors of mortality during CABG in hemodialysis (HD)-dependent patients. From the Japan Adult Cardiovascular Surgery Database, we compared 1,300 HD-dependent chronic renal failure patients with 18,387 non-HD patients who all underwent isolated CABG between January 2005 and December 2008. The operative mortality and mortality, including major morbidity, was 4.8% vs. 1.4% and 23.1% vs. 13.7% in the HD and non-HD groups, respectively. Preoperative predictors of operative mortality included age, chronic obstructive pulmonary disease, peripheral arterial disease, congestive heart failure, arrhythmia, preoperative inotropic agent requirement, New York Heart Association class IV, urgent or emergency operation, poor left ventricular function, aortic valve regurgitation (>2), and mitral valve regurgitation (>3). Postoperative predictors of operative mortality included stroke, infection, prolonged ventilation, pneumonia, heart block, and gastrointestinal complications. Compared with non-HD patients, CABG in HD patients was associated with high mortality and morbidity rates. An appropriate surgical strategy and careful perioperative assessment and management for prevention of respiratory and gastrointestinal complications might contribute to improved clinical outcomes after CABG in these patients.


Cyst formation in kidney via B-Raf signaling in the PKD2 transgenic mice.
The pathogenic mechanisms of human autosomal dominant polycystic kidney disease (ADPKD) have been well known to include the mutational inactivation of PKD2. Although haploinsufficiency and loss of heterozygosity at the Pkd2 locus can cause cyst formation in mice, polycystin-2 is frequently expressed in the renal cyst of human ADPKD, raising the possibility that deregulated activation of PKD2 may be associated with the cystogenesis of human ADPKD. To determine whether increased PKD2 expression is physiologically pathogenic, we generated PKD2-overexpressing transgenic mice. These mice developed typical renal cysts and an increase of proliferation and apoptosis, which are reflective of the human ADPKD phenotype. These manifestations were first observed at six months, and progressed with age. In addition, we found that ERK activation was induced by PKD2 overexpression via B-Raf signaling, providing a possible molecular mechanism of cystogenesis. In PKD2 transgenic mice, B-Raf/MEK/ERK sequential signaling was up-regulated. Additionally, the transgenic human polycystin-2 partially rescues the lethality of Pkd2 knock-out mice and therefore demonstrates that the transgene generated a functional product. Functional strengthening or deregulated activation of PKD2 may be a direct cause of ADPKD. The present study provides evidence for an in vivo role of overexpressed PKD2 in cyst formation. This transgenic mouse model should provide new insights into the pathogenic mechanism of human ADPKD.


A cell-penetrating peptide induces the self-reproduction of phospholipid vesicles: understanding the role of the bilayer rigidity.
Model lipid vesicles self-reproduce to generate unilamellar daughter vesicles in the presence of a cell-penetrating peptide. Fluorescence lifetime and anisotropy image analysis exquisitely capture the change in the lipid bilayer rigidity throughout the process, whereas a solvation dynamic study delineates the change in the dynamics of the encapsulated water inside the vesicles. Overall, our results provide an experimental underpinning of the role of the lipid bilayer structural changes in self-reproduction, which can exhibit great potential for interpreting the protein-membrane interaction in the emergence of life and for developing new therapeutic strategies.


Effect of human colostrum on interleukin-2 production and natural killer cell activity.
The effect of human colostrum on the production of interleukin-2 (IL-2) and on natural killer (NK) cell activity by peripheral blood mononuclear cells (PBMC) was investigated in 50 healthy women. At concentrations as low as 0.5%, human colostrum stimulated IL-2 production; at a higher concentration (10%), IL-2 secretion was inhibited. A time and dose dependent inhibitory effect of colostrum on NK cytotoxicity was also observed. This inhibition could be reversed by the addition of human recombinant IL-2 (hrIL-2). The stimulation of IL-2 production induced by human colostrum might compensate for its inhibitory effect on NK cell activity. These findings suggest an additional mechanism by which breast feeding may affect the neonatal immune system.


Alleviation of the pain of heel prick in preterm infants.
The hypothesis that the variability of physiological parameters may indicate pain or stress in the neonate was examined. Four parameters (heart rate, respiratory rate, transcutaneous oxygen tension, and carbon dioxide tension) were examined over a 2 minute epoch in response to a heel prick in an attempt to measure stress/pain in 35 preterm newborn infants (26-34 weeks' gestation) half of whom were receiving intensive care. The change in absolute values of these parameters did not discriminate a dummy procedure without prick from the actual procedure containing the prick (paired t test), but the variability of the parameters during an epoch showed significant discrimination. Three procedures were evaluated to reduce this distress using unpaired t test. The use of local anaesthetic cream was not successful. The components of the mixture cause vasoconstriction that would reduce blood flow to the heel and lead to more squeezing which is likely to be painful in the presence of tissue damage. A nurse comforting the infant with tactile and vocal stimulation was slightly helpful but the use of a spring loaded lance was most successful in reducing the distress. The use of spring loaded lances may be more humane for heel pricks.


Tyrosine phosphorylation of CRKL in Philadelphia+ leukemia.
The chimeric BCR/ABL protein is characteristic of Philadelphia (Ph)+ leukemia because it is the direct product of the Ph translocation and it has been shown to play a causal role in the genesis of leukemia. The BCR/ABL protein exhibits a deregulated tyrosine-kinase activity capable of phosphorylating different cellular substrates in vivo and in vitro. CRKL, an adaptor protein consisting of SH2 and SH3 domains in the absence of a catalytic domain, is one potential in vivo substrate of BCR/ABL. Previous experiments have shown that CRKL is phosphorylated on tyrosine in the chronic myelogenous leukemia (CML) cell line K562 and that CRKL is a substrate for ABL and for BCR/ABL in COS-1 cells. In the current study, we show that in peripheral blood cells a direct correlation exists between the presence of BCR/ABL and the phosphorylation status of CRKL. In Ph- peripheral blood cells, CRKL is present only in the nonphosphorylated form. In contrast, all BCR/ABL+ CML and acute lymphoblastic leukemia patient samples examined showed clear tyrosine-phosphorylation of CRKL. This result strongly suggests that CRKL is a biologically significant substrate for BCR/ABL and is likely to play a major role in the development of Ph+ leukemia.


Impact of intrauterine growth restriction on long-term health.
Intrauterine growth restriction (IUGR) is responsible for the higher rates of fetal, perinatal, and neonatal morbidity and mortality. This review details the IUGR risk factors, its short and long-term sequel, the mechanism underlying the long-term consequences, and the strategies to tackle IUGR burden. Short-term consequences of IUGR involve metabolic, thermal, and hematological disturbances leading to morbidities. Long term consequences due to changes in the fetal nutritional environment is associated with increased risk of developing metabolic syndrome and cardiovascular disease, systolic hypertension, obesity, insulin resistance, and diabetes type II in adulthood. There are no effective therapies to reverse IUGR, and antenatal management is aimed at determining the ideal time and mode of delivery. In order to prevent complications associated with IUGR, it is important to first detect the condition and institute appropriate surveillance to assess fetal well-being coupled with suitable intervention in case of fetal distress. Reliable prediction of IUGR may be achieved by combining clinical risk factors with Doppler abnormalities, fetal growth, and biomarkers. If this can be achieved, there is potential to reduce future perinatal morbidity, mortality and long-term consequences, but steps geared toward the prevention of IUGR are of unparalleled importance.


Binding and nuclear relocalization of protein kinase R by human cytomegalovirus TRS1.
The human cytomegalovirus (HCMV) TRS1 and IRS1 genes block the phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha) and the consequent shutoff of cellular protein synthesis that occur during infection with vaccinia virus (VV) deleted of the double-stranded RNA binding protein gene E3L (VVDeltaE3L). To further define the underlying mechanism, we first evaluated the effect of pTRS1 on protein kinase R (PKR), the double-stranded RNA (dsRNA)-dependent eIF2alpha kinase. Immunoblot analyses revealed that pTRS1 expression in the context of a VVDeltaE3L recombinant decreased levels of PKR in the cytoplasm and increased its levels in the nucleus of infected cells, an effect not seen with wild-type VV or a VVDeltaE3L recombinant virus expressing E3L. This effect of pTRS1 was confirmed by visualizing the nuclear relocalization of PKR-EGFP expressed by transient transfection. PKR present in both the nuclear and cytoplasmic fractions was nonphosphorylated, indicating that it was unactivated when TRS1 was present. PKR also accumulated in the nucleus during HCMV infection as determined by indirect immunofluorescence and immunoblot analysis. Binding assays revealed that pTRS1 interacted with PKR in mammalian cells and in vitro. This interaction required the same carboxy-terminal region of pTRS1 that is necessary to rescue VVDeltaE3L replication in HeLa cells. The carboxy terminus of pIRS1 was also required for rescue of VVDeltaE3L and for mediating an interaction of pIRS1 with PKR. These results suggest that these HCMV genes directly interact with PKR and inhibit its activation by sequestering it in the nucleus, away from both its activator, cytoplasmic dsRNA, and its substrate, eIF2alpha.


Alternative mRNA Splicing in Three Venom Families Underlying a Possible Production of Divergent Venom Proteins of the Habu Snake, Protobothrops flavoviridis.
Snake venoms are complex mixtures of toxic proteins encoded by various gene families that function synergistically to incapacitate prey. A huge repertoire of snake venom genes and proteins have been reported, and alternative splicing is suggested to be involved in the production of divergent gene transcripts. However, a genome-wide survey of the transcript repertoire and the extent of alternative splicing still remains to be determined. In this study, the comprehensive analysis of transcriptomes in the venom gland was achieved by using PacBio sequencing. Extensive alternative splicing was observed in three venom protein gene families, metalloproteinase (MP), serine protease (SP), and vascular endothelial growth factors (VEGF). Eleven MP and SP genes and a VEGF gene are expressed as a total of 81, 61, and 8 transcript variants, respectively. In the MP gene family, individual genes are transcribed into different classes of MPs by alternative splicing. We also observed trans-splicing among the clustered SP genes. No other venom genes as well as non-venom counterpart genes exhibited alternative splicing. Our results thus indicate a potential contribution of mRNA alternative and trans-splicing in the production of highly variable transcripts of venom genes in the habu snake.


Evidence for novel epigenetic marks within plants.
Variation in patterns of gene expression can result from modifications in the genome that occur without a change in the sequence of the DNA; such modifications include methylation of cytosine to generate 5-methylcytosine (5mC) resulting in the generation of heritable epimutation and novel epialleles. This type of non-sequence variation is called epigenetics. The enzymes responsible for generation of such DNA modifications in mammals are named DNA methyltransferases (DNMT) including DNMT1, DNMT2 and DNMT3. The later stages of oxidations to these modifications are catalyzed by Ten Eleven Translocation (TET) proteins, which contain catalytic domains belonging to the 2-oxoglutarate dependent dioxygenase family. In various mammalian cells/tissues including embryonic stem cells, cancer cells and brain tissues, it has been confirmed that these proteins are able to induce the stepwise oxidization of 5-methyl cytosine to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and finally 5-carboxylcytosine (5caC). Each stage from initial methylation until the end of the DNA demethylation process is considered as a specific epigenetic mark that may regulate gene expression. This review discusses controversial evidence for the presence of such oxidative products, particularly 5hmC, in various plant species. Whereas some reports suggest no evidence for enzymatic DNA demethylation, other reports suggest that the presence of oxidative products is followed by the active demethylation and indicate the contribution of possible TET-like proteins in the regulation of gene expression in plants. The review also summarizes the results obtained by expressing the human TET conserved catalytic domain in transgenic plants.


The use of fluorescence correlation spectroscopy to characterize the molecular mobility of fluorescently labelled G protein-coupled receptors.
The membranes of living cells have been shown to be highly organized into distinct microdomains, which has spatial and temporal consequences for the interaction of membrane bound receptors and their signalling partners as complexes. Fluorescence correlation spectroscopy (FCS) is a technique with single cell sensitivity that sheds light on the molecular dynamics of fluorescently labelled receptors, ligands or signalling complexes within small plasma membrane regions of living cells. This review provides an overview of the use of FCS to probe the real time quantification of the diffusion and concentration of G protein-coupled receptors (GPCRs), primarily to gain insights into ligand-receptor interactions and the molecular composition of signalling complexes. In addition we document the use of photon counting histogram (PCH) analysis to investigate how changes in molecular brightness (ε) can be a sensitive indicator of changes in molecular mass of fluorescently labelled moieties.


Rapid response to selection, competitive release and increased transmission potential of artesunate-selected Plasmodium chabaudi malaria parasites.
The evolution of drug resistance, a key challenge for our ability to treat and control infections, depends on two processes: de-novo resistance mutations, and the selection for and spread of resistant mutants within a population. Understanding the factors influencing the rates of these two processes is essential for maximizing the useful lifespan of drugs and, therefore, effective disease control. For malaria parasites, artemisinin-based drugs are the frontline weapons in the fight against disease, but reports from the field of slower parasite clearance rates during drug treatment are generating concern that the useful lifespan of these drugs may be limited. Whether slower clearance rates represent true resistance, and how this provides a selective advantage for parasites is uncertain. Here, we show that Plasmodium chabaudi malaria parasites selected for resistance to artesunate (an artemisinin derivative) through a step-wise increase in drug dose evolved slower clearance rates extremely rapidly. In single infections, these slower clearance rates, similar to those seen in the field, provided fitness advantages to the parasite through increased overall density, recrudescence after treatment and increased transmission potential. In mixed infections, removal of susceptible parasites by drug treatment led to substantial increases in the densities and transmission potential of resistant parasites (competitive release). Our results demonstrate the double-edged sword for resistance management: in our initial selection experiments, no parasites survived aggressive chemotherapy, but after selection, the fitness advantage for resistant parasites was greatest at high drug doses. Aggressive treatment of mixed infections resulted in resistant parasites dominating the pool of gametocytes, without providing additional health benefits to hosts. Slower clearance rates can evolve rapidly and can provide a strong fitness advantage during drug treatment in both single and mixed strain infections.


Survival at extreme altitude: protective effect of increased hemoglobin-oxygen affinity.
Decreased hemoglobin-oxygen affinity is thought to be of adaptive value to humans and nonindigenous animals at high altitude. To test this, hemoglobin-oxygen affinity was modified by carbamoylation of hemoglobin in rats. Exposure of control (low oxygen affinity) and experimental (high oxygen affinity) animals to a pressure equivalent to high altitude revealed that increased, rather than decreased, hemoglobin-oxygen affinity will permit survival at greatly reduced environmental oxygen pressures.


Syk-dependent actin dynamics regulate endocytic trafficking and processing of antigens internalized through the B-cell receptor.
Antigen binding to the B-cell receptor (BCR) induces multiple signaling cascades that ultimately lead to B lymphocyte activation. In addition, the BCR regulates the key trafficking events that allow the antigen to reach endocytic compartments devoted to antigen processing, i.e., that are enriched for major histocompatibility factor class II (MHC II) and accessory molecules such as H2-DM. Here, we analyze the role in antigen processing and presentation of the tyrosine kinase Syk, which is activated upon BCR engagement. We show that convergence of MHC II- and H2-DM-containing compartments with the vesicles that transport BCR-uptaken antigens is impaired in cells lacking Syk activity. This defect in endocytic trafficking compromises the ability of Syk-deficient cells to form MHC II-peptide complexes from BCR-internalized antigens. Altered endocytic trafficking is associated to a failure of Syk-deficient cells to properly reorganize their actin cytoskeleton in response to BCR engagement. We propose that, by modulating the actin dynamics induced upon BCR stimulation, Syk regulates the positioning and transport of the vesicles that carry the molecules required for antigen processing and presentation.


Reproductive characteristics modify the association between global DNA methylation and breast cancer risk in a population-based sample of women.
DNA methylation has been implicated in breast cancer aetiology, but little is known about whether reproductive history and DNA methylation interact to influence carcinogenesis. This study examined modification of the association between global DNA methylation and breast cancer risk by reproductive characteristics. A population-based case-control study assessed reproductive history in an interviewer-administered questionnaire. Global DNA methylation was measured from white blood cell DNA using luminometric methylation assay (LUMA) and pyrosequencing assay (long interspersed elements-1 (LINE-1). We estimated adjusted odds ratios (ORs) and 95% confidence intervals (CIs) among 1 070 breast cancer cases and 1 110 population-based controls. Effect modification was assessed on additive and multiplicative scales. LUMA methylation was associated with elevated breast cancer risk across all strata (comparing the highest to the lowest quartile), but estimates were higher among women with age at menarche ≤12 years (OR = 2.87, 95%CI = 1.96-4.21) compared to >12 years (OR = 1.66, 95%CI = 1.20-2.29). We observed a 2-fold increase in the LUMA methylation-breast cancer association among women with age at first birth >23 years (OR = 2.62, 95%CI = 1.90-3.62) versus ≤23 years (OR = 1.32, 95% CI = 0.84-2.05). No modification was evident for parity or lactation. Age at menarche and age at first birth may be modifiers of the association between global DNA methylation and breast cancer risk.


Prevalence and Risk Factors of Violence by Psychiatric Acute Inpatients: A Systematic Review and Meta-Analysis.
Violence in acute psychiatric wards affects the safety of other patients and the effectiveness of treatment. However, there is a wide variation in reported rates of violence in acute psychiatric wards. To use meta-analysis to estimate the pooled rate of violence in published studies, and examine the characteristics of the participants, and aspects of the studies themselves that might explain the variation in the reported rates of violence (moderators). Systematic meta-analysis of studies published between January 1995 and December 2014, which reported rates of violence in acute psychiatric wards of general or psychiatric hospitals in high-income countries. Of the 23,972 inpatients described in 35 studies, the pooled proportion of patients who committed at least one act of violence was 17% (95% confidence interval (CI) 14-20%). Studies with higher proportions of male patients, involuntary patients, patients with schizophrenia and patients with alcohol use disorder reported higher rates of inpatient violence. The findings of this study suggest that almost 1 in 5 patients admitted to acute psychiatric units may commit an act of violence. Factors associated with levels of violence in psychiatric units are similar to factors that are associated with violence among individual patients (male gender, diagnosis of schizophrenia, substance use and lifetime history of violence).


The effect of iron fortification on the fatty acid composition of plasma and erythrocyte membranes in primary school children with and without iron-deficiency.
An intervention study was designed to evaluate the fatty acid (FA) status of children aged 6-11 years before and after iron fortification. Iron deficient (ID) and matched controls without ID (n = 30) were selected. All children received soup (160 mL) fortified with 20 mg iron and 100 mg vitamin C for 15 weeks on school days. Measurements before and after intervention included dietary intake, haematological and iron status and FA composition of plasma and erythrocyte membranes (EMBs). The prevalence of low plasma ferritin concentration and transferrin saturation decreased in the ID children by 40% and 56%, respectively, with intervention. Plasma FAs reflected dietary FA intake. In comparison with controls, the ID group presented with increased percentage total saturated FAs (SFAs; p = 0.0002) in their EMB phosphatidylcholine (PC) and reduced percentage total polyunsaturated FAs (PUFAs; p = 0.0037) before intervention. Lower total n-3 FAs (p = 0.0070) including eicosapentaenoic acid (EPA; p = 0.0034), docosapentaenoic acid (DPA; p = 0.0048) and docosahexaenoic acid (DHA; p = 0.0058) were observed in the ID group. The EMB phosphatidylethanolamine (PEA) of the ID children presented with lower percentages of alpha-linolenic acid (ALA; p = 0.0001), EPA (p = 0.0051) and DHA (p = 0.0084) compared to controls before intervention. Iron intervention was associated with an increase (p < 0,05) in the percentage of n-3 FAs in the EMB-PC and -PEA of the ID group to percentages comparable to that in the control group. It appears that iron status can influence FA metabolism of specific n-3 FAs in the EMBs of young children.


HOY1, a homeo gene required for hyphal formation in Yarrowia lipolytica.
The dimorphic fungus Yarrowia lipolytica grows to form hyphae either in rich media or in media with GlcNAc as a carbon source. A visual screening, called FIL (filamentation minus), for Y. lipolytica yeast growth mutants has been developed. The FIL screen was used to identify three Y. lipolytica genes that abolish hypha formation in all media assayed. Y. lipolytica HOY1, a gene whose deletion prevents the yeast-hypha transition both in liquid and solid media, was characterized. HOY1 is predicted to encode a 509-amino-acid protein with a homeodomain homologous to that found in the chicken Hox4.8 gene. Analysis of the protein predicts a nuclear location. These observations suggest that Hoy1p may function as a transcriptional regulatory protein. In disrupted strains, reintroduction of HOY1 restored the capacity for hypha formation. Northern blot hybridization revealed the HOY1 transcript to be approximately 1.6 kb. Expression of this gene was detected when Y. lipolytica grew as a budding yeast, but an increase in its expression was observed by 1 h after cells had been induced to form hyphae. The possible functions of HOY1 in hyphal growth and the uses of the FIL screen to identify morphogenetic regulatory genes from heterologous organisms are discussed.


Osteoblasts mediate the adverse effects of glucocorticoids on fuel metabolism.
Long-term glucocorticoid treatment is associated with numerous adverse outcomes, including weight gain, insulin resistance, and diabetes; however, the pathogenesis of these side effects remains obscure. Glucocorticoids also suppress osteoblast function, including osteocalcin synthesis. Osteocalcin is an osteoblast-specific peptide that is reported to be involved in normal murine fuel metabolism. We now demonstrate that osteoblasts play a pivotal role in the pathogenesis of glucocorticoid-induced dysmetabolism. Osteoblast-targeted disruption of glucocorticoid signaling significantly attenuated the suppression of osteocalcin synthesis and prevented the development of insulin resistance, glucose intolerance, and abnormal weight gain in corticosterone-treated mice. Nearly identical effects were observed in glucocorticoid-treated animals following heterotopic (hepatic) expression of both carboxylated and uncarboxylated osteocalcin through gene therapy, which additionally led to a reduction in hepatic lipid deposition and improved phosphorylation of the insulin receptor. These data suggest that the effects of exogenous high-dose glucocorticoids on insulin target tissues and systemic energy metabolism are mediated, at least in part, through the skeleton.


Short-term hematologic and hemodynamic effects of osteopathic lymphatic techniques: a pilot crossover trial.
Research into the physiologic effects of osteopathic lymphatic techniques has been somewhat limited. To assess the short-term hematologic and hemodynamic effects of a comprehensive lymphatic treatment protocol. Randomized crossover design that included 10-minute lymphatic treatment and rest (control) protocols delivered 1 week apart for a small pilot group of healthy men (N=15). At baseline, albumin, hematocrit, hemoglobin and platelet count , total protein, and white blood cell count were measured, as were systolic and diastolic blood pressure. All measures were repeated 20, 50, and 80 minutes after baseline data were gathered. Significant condition x time interaction effects were observed, indicating a decrease in platelet counts and an increase in diastolic blood pressure after the lymphatic treatment protocol [corrected]. Statistically significant differences by time were observed in all hemotologic measures and in systolic blood pressure. No adverse events or complications from the treatment protocol were observed in this population. Lymphatic techniques may decrease platelet counts and increase diastolic blood pressure during the first hour after treatment.


Calcium and orthophosphate deposits in vitro do not imply osteoblast-mediated mineralization: mineralization by betaglycerophosphate in the absence of osteoblasts.
It has been shown in several laboratories that addition of beta-glycerophosphate (beta GP), a substrate for alkaline phosphatase (AP), to cultured osteoblast-like cells induces deposition of orthophosphate (Pi) and Ca within seven days. Even though this effect is regarded as an in vitro model of bone mineralization, it is not known whether it is specific for osteoblasts. We have, therefore, studied the amounts of Pi and Ca deposited after seven days with 10 mM beta GP in culture wells containing confluent cultures of osteoblast-like cells (OB) derived from human trabecular bone explants, human skin fibroblasts (SF), or culture medium alone (MED). Ox liver AP at an activity considerably greater than the endogenous AP activity of the cells, but comparable with that of other osteoblast models, was added to ensure a similar rate of Pi generation from beta GP in all wells. beta GP was converted quantitatively to Pi within seven days, leading to a nonphysiological 10-fold increase in the Pi concentration in the culture medium. After thorough rinsing on day seven, the OB and SF wells contained deposits of Pi and Ca, but the amounts were comparable for the two cell types. Smaller, but significant, amounts of Pi and Ca were also detectable even in rinsed MED wells. This suggests that the detection of such deposits in beta GP experiments cannot necessarily be interpreted as a specific property of osteoblast cultures in vitro, and may simply reflect the presence of AP.(ABSTRACT TRUNCATED AT 250 WORDS)


Lifetime alcohol consumption and severity in alcohol dependence syndrome.
Alcohol dependence syndrome is a major public health issue globally and is responsible for significant morbidity and mortality. The total dose of alcohol consumed has been linked to liver diseases, pancreatitis, and other alcohol-related medical consequences. However, this has not been studied in relation to severity of dependence; although it is well known that alcohol causes neuronal damage, which in turn potentiates dependence. Thus, there is a need to study the relationship between the amount of alcohol consumed and severity of dependence. A total of 165 consecutive cases of alcohol dependence syndrome were studied in a General Hospital Psychiatry Unit at a tertiary care hospital. Addiction Severity Index (ASI) was used to evaluate the severity of alcohol dependence, and Life Time Alcohol Consumption (LTAC) was evaluated by taking careful history. Correlation coefficients were calculated between ASI and LTAC. Group differences were analyzed using t-test. There was a significant correlation between ASI and LTAC (r = 0.162, P = 0.032), which was highly significant in the subgroup without medical complications (r = 0.250, P = 0.003). A similar correlation in the medical complications subgroup was not significant. Lifetime alcohol consumption co-related with the severity of alcohol dependence, particularly in those presenting without medical complications (i.e., those with behavioral and social consequences, and injuries).


Impact of pile-driving on Hector's dolphin in Lyttelton Harbour, New Zealand.
Several dolphin species occur close inshore and in harbours, where underwater noise generated by pile-driving used in wharf construction may constitute an important impact. Such impacts are likely to be greatest on species such as the endangered Hector's dolphin (Cephalorhynchus hectori), which has small home ranges and uses this habitat type routinely. Using automated echolocation detectors in Lyttelton Harbour (New Zealand), we studied the distribution of Hector's dolphins using a gradient sampling design over 92 days within which pile-driving occurred on 46 days. During piling operations, dolphin positive minutes per day decreased at the detector closest to the piling but increased at the mid-harbour detector. Finer-grained analyses showed that close to the piling operation, detections decreased with increasing sound exposure level, that longer piling events were associated with longer reductions in detections, and that effects were long-lasting - detection rates took up to 83 h to return to pre-piling levels.


Can the Dying Phase Be Masked by the Use of Dexamethasone? A Case Report.
Recognition of the dying phase, i.e., the period during which death is expected to occur within hours or days, is important because it enables marking the imminence of death, informing the patient and his relatives, and adjusting care where needed. Careful communication about a patient's limited prognosis prepares patients and their family for impending death and saying goodbye. The authors describe two cases of patients dying a relatively unexpected death in a hospice, which is uncommon in this setting. These unexpected deaths had a severe impact on the relatives and on the professional care team. Both patients used dexamethasone. The authors postulate that there is a relationship between the use of dexamethasone and difficulty in recognizing the dying phase. Dexamethasone can make patients feel better, increase their appetite, and influence the stress response. These effects could mask the signs of impending death, such as "being bedbound," "only drinking sips," and "being subcomatose." Review of the literature yielded no articles that support or reject this hypothesis. Because dexamethasone is used regularly in the palliative phase of a chronic disease, there is a need for further exploration of its effects in the dying phase.


Anomalous Nonlinear Dynamics Behavior of Fractional Viscoelastic Beams.
Fractional models and their parameters are sensitive to intrinsic microstructural changes in anomalous materials. We investigate how such physics-informed models propagate the evolving anomalous rheology to the nonlinear dynamics of mechanical systems. In particular, we study the vibration of a fractional, geometrically nonlinear viscoelastic cantilever beam, under base excitation and free vibration, where the viscoelasticity is described by a distributed-order fractional model. We employ Hamilton's principle to obtain the equation of motion with the choice of specific material distribution functions that recover a fractional Kelvin-Voigt viscoelastic model of order α. Through spectral decomposition in space, the resulting time-fractional partial differential equation reduces to a nonlinear time-fractional ordinary differential equation, where the linear counterpart is numerically integrated through a direct L1-difference scheme. We further develop a semi-analytical scheme to solve the nonlinear system through a method of multiple scales, yielding a cubic algebraic equation in terms of the frequency. Our numerical results suggest a set of α-dependent anomalous dynamic qualities, such as far-from-equilibrium power-law decay rates, amplitude super-sensitivity at free vibration, and bifurcation in steady-state amplitude at primary resonance.


Investigating Sex Bias in the AQ-10: A Replication Study.
There are concerns that females with autism spectrum disorders (ASD) may be underidentified because of factors such as better camouflaging and poorer recognition of the signs of ASD in females. One stage at which females may be underidentified is during screening. In this study, we, therefore, evaluated whether the autism spectrum quotient (AQ-10), a brief recommended screening instrument for ASD in adults suspected of having ASD, showed any evidence of underestimating symptoms in females. Our results broadly replicate those of an earlier study in finding no strong evidence that the AQ-10 is biased against females. However, to achieve better performance in females, we suggest that one item be replaced with an item measuring more "female" manifestations of ASD.


Symbiotic and antibiotic interactions between gut commensal microbiota and host immune system.
The human gut commensal microbiota forms a complex population of microorganisms that survive by maintaining a symbiotic relationship with the host. Amongst the metabolic benefits it brings, formation of adaptive immune system and maintenance of its homeostasis are functions that play an important role. This review discusses the integral elements of commensal microbiota that stimulate responses of different parts of the immune system and lead to health or disease. It aims to establish conditions and factors that contribute to gut commensal microbiota's transformation from symbiotic to antibiotic relationship with human. We suggest that the host-microbiota relationship has been evolved to benefit both parties and any changes that may lead to disease, are not due to unfriendly properties of the gut microbiota but due to host genetics or environmental changes such as diet or infection.


Structure of the ent-Copalyl Diphosphate Synthase PtmT2 from Streptomyces platensis CB00739, a Bacterial Type II Diterpene Synthase.
Terpenoids are the largest and most structurally diverse family of natural products found in nature, yet their presence in bacteria is underappreciated. The carbon skeletons of terpenoids are generated through carbocation-dependent cyclization cascades catalyzed by terpene synthases (TSs). Type I and type II TSs initiate cyclization via diphosphate ionization and protonation, respectively, and protein structures of both types are known. Most plant diterpene synthases (DTSs) possess three α-helical domains (αβγ), which are thought to have arisen from the fusion of discrete, ancestral bacterial type I TSs (α) and type II TSs (βγ). Type II DTSs of bacterial origin, of which there are no structurally characterized members, are a missing piece in the structural evolution of TSs. Here, we report the first crystal structure of a type II DTS from bacteria. PtmT2 from Streptomyces platensis CB00739 was verified as an ent-copalyl diphosphate synthase involved in the biosynthesis of platensimycin and platencin. The crystal structure of PtmT2 was solved at a resolution of 1.80 Å, and docking studies suggest the catalytically active conformation of geranylgeranyl diphosphate (GGPP). Site-directed mutagenesis confirmed residues involved in binding the diphosphate moiety of GGPP and identified DxxxxE as a potential Mg(2+)-binding motif for type II DTSs of bacterial origin. Finally, both the shape and physicochemical properties of the active sites are responsible for determining specific catalytic outcomes of TSs. The structure of PtmT2 fundamentally advances the knowledge of bacterial TSs, their mechanisms, and their role in the evolution of TSs.


Acquisition of membrane lipids by differentiating glyoxysomes: role of lipid bodies.
Glyoxysomes in cotyledons of cotton (Gossypium hirsutum, L.) seedlings enlarge dramatically within 48 h after seed imbibition (Kunce, C.M., R.N. Trelease, and D.C. Doman. 1984. Planta (Berl.). 161:156-164) to effect mobilization of stored cotton-seed oil. We discovered that the membranes of enlarging glyoxysomes at all stages examined contained a large percentage (36-62% by weight) of nonpolar lipid, nearly all of which were triacylglycerols (TAGs) and TAG metabolites. Free fatty acids comprised the largest percentage of these nonpolar lipids. Six uncommon (and as yet unidentified) fatty acids constituted the majority (51%) of both the free fatty acids and the fatty acids in TAGs of glyoxysome membranes; the same six uncommon fatty acids were less than 7% of the acyl constituents in TAGs extracted from cotton-seed storage lipid bodies. TAGs of lipid bodies primarily were composed of palmitic, oleic, and linoleic acids (together 70%). Together, these three major storage fatty acids were less than 10% of both the free fatty acids and fatty acids in TAGs of glyoxysome membranes. Phosphatidylcholine (PC) and phosphatidylethanolamine (PE) constituted a major portion of glyoxysome membrane phospholipids (together 61% by weight). Pulse-chase radiolabeling experiments in vivo clearly demonstrated that 14C-PC and 14C-PE were synthesized from 14C-choline and 14C-ethanolamine, respectively, in ER of cotyledons, and then transported to mitochondria; however, these lipids were not transported to enlarging glyoxysomes. The lack of ER involvement in glyoxysome membrane phospholipid synthesis, and the similarities in lipid compositions between lipid bodies and membranes of glyoxysomes, led us to formulate and test a new hypothesis whereby lipid bodies serve as the dynamic source of nonpolar lipids and phospholipids for membrane expansion of enlarging glyoxysomes. In a cell-free system, 3H-triolein (TO) and 3H-PC were indeed transferred from lipid bodies to glyoxysomes. 3H-PC, but not 3H-TO, also was transferred to mitochondria in vitro. The amount of lipid transferred increased linearly with respect to time and amount of acceptor organelle protein, and transfer occurred only when lipid body membrane proteins were associated with the donor lipid bodies. 3H-TO was transferred to and incorporated into glyoxysome membranes, and then hydrolyzed to free fatty acids. 3H-PC was transferred to and incorporated into glyoxysome and mitochondria membranes without subsequent hydrolysis. Our data are inconsistent with the hypothesis that ER contributes membrane lipids to glyoxysomes during postgerminative seedling growth.(ABSTRACT TRUNCATED AT 400 WORDS)


Journey of mesenchymal stem cells for homing: strategies to enhance efficacy and safety of stem cell therapy.
Human mesenchymal stem cells (MSCs) communicate with other cells in the human body and appear to "home" to areas of injury in response to signals of cellular damage, known as homing signals. This review of the state of current research on homing of MSCs suggests that favorable cellular conditions and the in vivo environment facilitate and are required for the migration of MSCs to the site of insult or injury in vivo. We review the current understanding of MSC migration and discuss strategies for enhancing both the environmental and cellular conditions that give rise to effective homing of MSCs. This may allow MSCs to quickly find and migrate to injured tissues, where they may best exert clinical benefits resulting from improved homing and the presence of increased numbers of MSCs.


Age-related changes in object processing and contextual binding revealed using fMR adaptation.
Using fMR adaptation, we studied the effects of aging on the neural processing of passively viewed naturalistic pictures composed of a prominent object against a background scene. Spatially distinct neural regions showing specific patterns of adaptation to objects, background scenes, and contextual integration (binding) were identified in young adults. Older adults did not show adaptation responses corresponding to binding in the medial-temporal areas. They also showed an adaptation deficit for objects whereby their lateral occipital complex (LOC) did not adapt to repeated objects in the context of a changing background. The LOC could be activated, however, when objects were presented without a background. Moreover, the adaptation deficit for objects viewed against backgrounds was reversed when elderly subjects were asked to attend to objects while viewing these complex pictures. These findings suggest that the elderly have difficulty with simultaneous processing of objects and backgrounds that, in turn, could contribute to deficient contextual binding.


Microbiome sharing between children, livestock and household surfaces in western Kenya.
The gut microbiome community structure and development are associated with several health outcomes in young children. To determine the household influences of gut microbiome structure, we assessed microbial sharing within households in western Kenya by sequencing 16S rRNA libraries of fecal samples from children and cattle, cloacal swabs from chickens, and swabs of household surfaces. Among the 156 households studied, children within the same household significantly shared their gut microbiome with each other, although we did not find significant sharing of gut microbiome across host species or household surfaces. Higher gut microbiome diversity among children was associated with lower wealth status and involvement in livestock feeding chores. Although more research is necessary to identify further drivers of microbiota development, these results suggest that the household should be considered as a unit. Livestock activities, health and microbiome perturbations among an individual child may have implications for other children in the household.


Microscopic origin of the chiroptical response of optical media.
The potential for enhancing the optical activity of natural chiral media using engineered nanophotonic components has been central in the quest toward developing next-generation circular-dichroism spectroscopic techniques. Through confinement and manipulation of optical fields at the nanoscale, ultrathin optical elements have enabled a path toward achieving order-of-magnitude enhancements in the chiroptical response. Here, we develop a model framework to describe the underlying physics governing the origin of the chiroptical response in optical media. The model identifies optical activity to originate from electromagnetic coupling to the hybridized eigenstates of a coupled electron-oscillator system, whereas differential absorption of opposite handedness light, though resulting in a far-field chiroptical response, is shown to have incorrectly been identified as optical activity. We validate the model predictions using experimental measurements and show them to also be consistent with observations in the literature. The work provides a generalized framework for the design and study of chiroptical systems.


Optimized labeling of membrane proteins for applications to super-resolution imaging in confined cellular environments using monomeric streptavidin.
Recent progress in super-resolution imaging (SRI) has created a strong need to improve protein labeling with probes of small size that minimize the target-to-label distance, increase labeling density, and efficiently penetrate thick biological tissues. This protocol describes a method for labeling genetically modified proteins incorporating a small biotin acceptor peptide with a 3-nm fluorescent probe, monomeric streptavidin. We show how to express, purify, and conjugate the probe to organic dyes with different fluorescent properties, and how to label selectively biotinylated membrane proteins for SRI techniques (point accumulation in nanoscale topography (PAINT), stimulated emission depletion (STED), stochastic optical reconstruction microscopy (STORM)). This method is complementary to the previously described anti-GFP-nanobody/SNAP-tag strategies, with the main advantage being that it requires only a short 15-amino-acid tag, and can thus be used with proteins resistant to fusion with large tags and for multicolor imaging. The protocol requires standard molecular biology/biochemistry equipment, making it easily accessible for laboratories with only basic skills in cell biology and biochemistry. The production/purification/conjugation steps take ∼5 d, and labeling takes a few minutes to an hour.


[Influence of hyponutrition on occurrence of complications and mortality among cirrhosis patients].
Nutrient Energy Malnutrition (NEM) is a common feature among liver cirrhosis (LC) patients, and might constitute a negative predictor of patient's survival. A prospective descriptive study was carried out in order to assess the influence of LC patient's nutritional status on the occurrence of complications and mortality in a tertiary care health institution in Havana City (Cuba). One-hundred twenty-one patients were recruited for the study (Women: 52.9%; Younger than 60 years: 59.0%; Whites: 90.0%). C virus infection, alcoholism, and B virus infection accounted for 71.0% of LC cases. Ninety-two percents of the patients had less than 5 yearse of diagnosis. Subjective Global Assessment (SGA) of nutritional status returned a 45.0% of malnutrition. Malnutrition was associated with disease progression: twenty-one percent of A Child-Pugh Stage patients was malnourished, but this rate increased to become 90.0% among those in C stage. Fat and skeletal muscle compartments were reduced in 36.0 and 37.0% of the patients, respectively. Complications affected 44.6% of the patients. Sixty percent of them were malnourished (p <0.01). One-year mortality rate was 11.0%. Ninety-two percent of deceased patients were malnourished (p < 0.01). LC malnourished patients have greater risk of complications and death as compared to well-nourished ones.


The Copenhagen cohort study on infant nutrition and growth: duration of breast feeding and influencing factors.
Duration of breast feeding was studied in 249 randomly chosen, healthy, term infants of Danish origin of which 80.7% participated. Breast feeding was initiated by 99.5% of the mothers. At 3, 6 and 9 months, 71%, 52% and 33%, respectively, were still breast feeding. Only 1 infant (0.5%) was exclusively breast fed beyond 7 months of age. In a Cox multiple regression analysis of factors influencing duration of breast feeding, we found a positive association with maternal education (p < 0.001) and age (p = 0.02) and a negative association with the amount of formula given at the maternity ward (p < 0.001). Six months after delivery, 79% of the mothers with higher school education (> or = 12 years) were still breast feeding, compared to 29% with a low school education (< or = 9 years). There is still a need for an increased effort to support mothers in breast feeding, focusing particularly on younger mothers with short school education. Formula supplements during the first days of life, given to 73% of the infants, were associated with a shorter duration of breast feeding and should be discouraged.


The use of coccygeal discs to study intervertebral disc metabolism.
There have been very few studies on the metabolism of the intervertebral disc. One possible reason is that lumbar discs suitable for in vitro synthesis studies are difficult to obtain. We have examined some properties of coccygeal discs to see if those discs are a suitable alternative. The properties measured appear similar to those of lumbar discs, with collagen content lowest in the nucleus and highest in the outer annulus, while hydration and glycosaminoglycan content were highest in the nucleus and of a similar level to those of canine lumbar discs (approximately 83% water). The discs were under mechanical load in vivo, resulting in an equilibrium swelling pressure of 0.25-0.3 MPa. The 35S-sulphate incorporation rate was 2-5 x 10(-5) mmol/g dry weight per hour in the inner annulus and nucleus, in comparison with 2-3 x 10(-5) mmol/g dry weight per hour in rabbit and canine lumbar discs. Coccygeal discs are a readily obtainable and cheap source of disc material, and they are easy to dissect out. Since the general properties of these discs are similar to those of lumbar discs, we suggest that they are a suitable model for the study of the metabolic and other properties of lumbar discs.