Triple-site radiotracer application in breast lymphoscintigraphy and sentinel node discordance.
Sentinel lymph node (SLN) biopsy (SLNB) has demonstrated accuracy in the axillary staging of breast cancer patients. Despite variability in selection criteria and technique, an SLN is consistently identified in approximately 96% of cases and in most series predicts the status of remaining axillary LNs in >95% of cases. The false-negative rate of sentinel lymph node biopsy (SLNB) was originally reported as 5%-10% (sensitivity 90%-95%), but improved rates are attainable by experienced surgeons. Radiolocalization with lymphoscintigraphy (LSG) increases SLN identification rates. LSG is a useful tool to establish the abnormal lymphatic drainage patterns and to detect the extra-axillary nodes, particularly internal mammary nodes. Despite controversy regarding the optimal injection method, studies have generally suggested high concordance between the various radiotracer application sites and axillary SLN identification. Discordant SLN identification would have implications for nodal staging as the true SLN might not be identified with individual injection techniques. In the current study, imaging from consecutive patients presenting for breast LSG over a-19 month period was retrospectively reviewed. Radiotracer application was performed with simultaneous injection of peritumoral, subcutaneous, and subareolar regions. This application method provided a mechanism to assess the LSG drainage patterns with a view to assessing injection site concordance and SLN identification rates. Data from 123 breast LSG patients were reviewed. Using our radiotracer technique, the axillary SLN identification rate was 98%. A single axillary node was detected in 110, two axillary nodes were detected in 10, and no axillary node was detected in three patients. Among those 10 patients in whom two axillary nodes were seen, at least two cases of discordant drainage occurred from different injection sites. This study demonstrates that different LSG injection sites can result in the identification of different axillary sentinel nodes although this appears to be a rare event. This finding may be of clinical importance if the true SLN is sought. In addition, the multisite injection technique appears to be an optimal method of axillary SLN identification, with high SLN detection rates.


Safety evaluation of the food enzyme endo-1,4-β-xylanase from a genetically modified Bacillus subtilis (strain LMG S-24584).
The food enzyme endo-1,4-β-xylanase (EC 220.127.116.11) is produced with the genetically modified Bacillus subtilis strain LMG S-24584 by Puratos N. V. The genetic modifications do not give rise to safety concerns. The Panel noted that, although the production strain was not detected in the food enzyme, recombinant DNA was present in all batches of the food enzyme tested. The food enzyme is intended to be used in baking processes. Based on the maximum use levels recommended for the baking processes and individual consumption data from the EFSA Comprehensive European Food Consumption Database, dietary exposure to the food enzyme-Total Organic Solids (TOS) was estimated to be up to 0.017 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rodents. A comparison of the no observed adverse effect level of 37 mg TOS/kg bw per day from this study with the dietary exposure results in a sufficiently high margin of exposure. The amino acid sequence of the food enzyme did not match those of known allergens. The Panel considered that, under the intended condition of use, the risk of allergic sensitisation and elicitation reactions upon dietary exposure to this food enzyme cannot be excluded, but the likelihood of such reactions occurring is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.


Impact of Muscle Measures on Outcome in Patients Receiving Endocrine Therapy for Metastatic Breast Cancer: Analysis of ECOG-ACRIN E2112.
Observational data investigating the relationship between body habitus and outcomes in breast cancer have been variable and inconsistent, largely centered in the curative setting and focused on weight-based metrics. This study evaluated the impact of muscle measures on outcomes in patients with metastatic breast cancer receiving endocrine-based therapy. Baseline CT scans were collected from ECOG-ACRIN E2112, a randomized phase III placebo-controlled study of exemestane with or without entinostat. A CT cross-sectional image at the L3 level was extracted to obtain skeletal muscle mass and attenuation. Low muscle mass (LMM) was defined as skeletal muscle index <41 cm2/m2 and low muscle attenuation (LMA) as muscle density <25 HU or <33 HU if overweight/obese by body mass index (BMI). Multivariable Cox proportional hazard models determined the association between LMM or LMA and progression-free survival (PFS) and overall survival (OS). Correlations between LMM, LMA, and patient-reported outcomes were determined using 2-sample t tests. Analyzable CT scans and follow-up data were available for 540 of 608 patients. LMM was present in 39% (n=212) of patients and LMA in 56% (n=301). Those with LMA were more likely to have obesity and worse performance status. LMM was not associated with survival (PFS hazard ratio [HR]: 1.13, P=.23; OS HR: 1.05, P=.68), nor was LMA (PFS HR: 1.01, P=.93; OS HR: 1.00, P=.99). BMI was not associated with survival. LMA, but not LMM, was associated with increased frequency of patient-reported muscle aches. Both low muscle mass and density are prevalent in patients with hormone receptor-positive metastatic breast cancer. Muscle measures correlated with obesity and performance status; however, neither muscle mass nor attenuation were associated with prognosis. Further work is needed to refine body composition measurements and select optimal cutoffs with meaningful endpoints in specific breast cancer populations, particularly those living with metastatic disease.


Spinal epidural hematoma.
Spinal epidural hematoma is a rare condition that usually presents with acute, severe pain at the location of the hemorrhage, with radiation to the extremities. It can rapidly develop to include progressive and severe neurologic deficit. The pathophysiology often remains unclear. However, epidural hematoma in the lumbar spine is best described as the result of internal rupture of the Batson vertebral venous plexus. Clinical evaluation of pain control and neurologic deficit is the most important tool in early diagnosis. Currently, MRI is the diagnostic method of choice. Regardless of the setting, symptomatic spinal epidural hematoma is typically managed with urgent surgical decompression of the spinal canal.


An integrated centrifugo-opto-microfluidic platform for arraying, analysis, identification and manipulation of individual cells.
In this work we present a centrifugal microfluidic system enabling highly efficient collective trapping and alignment of particles such as microbeads and cells, their multi-colour fluorescent detection and subsequent manipulation by optical tweezers. We demonstrate array-based capture and imaging followed by "cherry-picking" of individual particles, first for fluorescently labelled polystyrene (PS) beads and then for cells. Different cell lines are discriminated based on intracellular as well as surface-based markers.


Establishment of a yeast-based VLP platform for antigen presentation.
Chimeric virus-like particles (VLP) allow the display of foreign antigens on their surface and have proved valuable in the development of safe subunit vaccines or drug delivery. However, finding an inexpensive production system and a VLP scaffold that allows stable incorporation of diverse, large foreign antigens are major challenges in this field. In this study, a versatile and cost-effective platform for chimeric VLP development was established. The membrane integral small surface protein (dS) of the duck hepatitis B virus was chosen as VLP scaffold and the industrially applied and safe yeast Hansenula polymorpha (syn. Pichia angusta, Ogataea polymorpha) as the heterologous expression host. Eight different, large molecular weight antigens of up to 412 amino acids derived from four animal-infecting viruses were genetically fused to the dS and recombinant production strains were isolated. In all cases, the fusion protein was well expressed and upon co-production with dS, chimeric VLP containing both proteins could be generated. Purification was accomplished by a downstream process adapted from the production of a recombinant hepatitis B VLP vaccine. Chimeric VLP were up to 95% pure on protein level and contained up to 33% fusion protein. Immunological data supported surface exposure of the foreign antigens on the native VLP. Approximately 40 mg of chimeric VLP per 100 g dry cell weight could be isolated. This is highly comparable to values reported for the optimized production of human hepatitis B VLP. Purified chimeric VLP were shown to be essentially stable for 6 months at 4 °C. The dS-based VLP scaffold tolerates the incorporation of a variety of large molecular weight foreign protein sequences. It is applicable for the display of highly immunogenic antigens originating from a variety of pathogens. The yeast-based production system allows cost-effective production that is not limited to small-scale fundamental research. Thus, the dS-based VLP platform is highly efficient for antigen presentation and should be considered in the development of future vaccines.


Pathophysiology of insulin secretion in non-insulin-dependent diabetes mellitus.
The pathogenesis of the abnormal metabolic state in patients with non-insulin-dependent diabetes (NIDDM) is controversial. Even the term NIDDM stirs controversy because of the easily drawn inference that individuals with this form of diabetes do not need insulin treatment. Yet many patients with NIDDM are treated with insulin; some even develop hyperosmolar coma if not given insulin. Ketoacidosis, however, is very infrequent in this syndrome, implying that these patients are not dependent on insulin treatment to prevent mass mobilization of fatty acids and ketone bodies. The phrase noninsulin-dependent is therefore appropriate when used in this restricted fashion but inappropriate when used to imply adequacy of insulin secretion. The evaluation of the adequacy of islet function in this syndrome has been complex, since there is no standard of insulin output that can be defined for normal islets without specifying the physiologic setting under which the assessment has been made. For example, individuals with normal glucose levels but variable degrees of obesity have widely varying insulin secretion rates. Thus, the choice of controls is critical when comparing islet function in NIDDM to normal. In addition, the efficiency of the B-cell response to a challenge (e.g., oral glucose tolerance test) can markedly influence the magnitude of the stimulatory glucose level during the period of testing. For example, a subject with some impairment of insulin output will tend to become more hyperglycemic during the test. The hyperglycemia may then stimulate more insulin secretion so that the overall insulin output may appear equal to or even greater than that of a normal individual. In such a closed-loop system, strict control of input variables is necessary to evaluate whether or not insulin secretion is normal. As will be discussed, control of glucose level and other variables is seldom accomplished in dynamic glucose tolerance tests. As will be presented in this review, the development of appropriately controlled studies of islet function has provided convincing evidence that islet B-cell function is abnormal in patients with NIDDM. Since these studies are based on an understanding of normal islet function, normal islet B-cell physiology is discussed before pathophysiology. Finally, the implications of this analysis for the treatment of NIDDM with diet, hypoglycemic sulfonylureas, and insulin will be discussed.


Thermally activated TRPV3 channels.
TRPV3 is a temperature-sensitive transient receptor potential (TRP) ion channel. The TRPV3 protein functions as a Ca(2+)-permeable nonselective cation channel with six transmembrane domains forming a tetrameric complex. TRPV3 is known to be activated by warm temperatures, synthetic small-molecule chemicals, and natural compounds from plants. Its function is regulated by a variety of physiological factors including extracellular divalent cations and acidic pH, intracellular adenosine triphosphate, membrane voltage, and arachidonic acid. TRPV3 shows a broad expression pattern in both neuronal and non-neuronal tissues including epidermal keratinocytes, epithelial cells in the gut, endothelial cells in blood vessels, and neurons in dorsal root ganglia and CNS. TRPV3 null mice exhibit abnormal hair morphogenesis and compromised skin barrier function. Recent advances suggest that TRPV3 may play critical roles in inflammatory skin disorders, itch, and pain sensation. Thus, identification of selective TRPV3 activators and inhibitors could potentially lead to beneficial pharmacological interventions in several diseases. The intent of this review is to summarize our current knowledge of the tissue expression, structure, function, and mechanisms of activation of TRPV3.


Substance use comorbidity among veterans with posttraumatic stress disorder and other psychiatric illness.
There is considerable concern about the emergence of significant substance abuse among younger veterans of war in the Middle East, especially among those with Posttraumatic Stress Disorder (PTSD), but little information exists on the magnitude of this problem. Using national administrative data from the Department of Veterans Affairs (VA) (n = 1,001,996), we examined rates of diagnosed substance use disorders in Veterans who served in Iraq and Afghanistan diagnosed with PTSD compared to other psychiatric disorders; and compared rates among veterans of other service eras. Of VA patients with a selected mental disorder, 21.0% had a comorbid substance diagnosis. Veterans who served in the post-Vietnam era (VET) (1973-1991) had the highest rates of comorbidity. Logistic regression models indicated that veterans with each selected psychiatric diagnosis were significantly more likely to be dually diagnosed in comparison to veterans with PTSD; post-Vietnam veterans were significantly more likely to be dually diagnosed than veterans from other eras. Bipolar disorder and schizophrenia are most strongly associated with dual diagnosis in OEF/OIF (Operation Enduring Freedom/Operation Iraqi Freedom) veterans. There are high rates of substance use disorders among veterans with mental illness. The highest rates of comorbidity occur among those with bipolar disorder and schizophrenia; and in post-VET veterans.


Microbial ecology of deep-sea hypersaline anoxic basins.
Deep hypersaline anoxic basins (DHABs) are unique water bodies occurring within fractures at the bottom of the sea, where the dissolution of anciently buried evaporites created dense anoxic brines that are separated by a chemocline/pycnocline from the overlying oxygenated deep-seawater column. DHABs have been described in the Gulf of Mexico, the Mediterranean Sea, the Black Sea and the Red Sea. They are characterized by prolonged historical separation of the brines from the upper water column due to lack of mixing and by extreme conditions of salinity, anoxia, and relatively high hydrostatic pressure and temperatures. Due to these combined selection factors, unique microbial assemblages thrive in these polyextreme ecosystems. The topological localization of the different taxa in the brine-seawater transition zone coupled with the metabolic interactions and niche adaptations determine the metabolic functioning and biogeochemistry of DHABs. In particular, inherent metabolic strategies accompanied by genetic adaptations have provided insights on how prokaryotic communities can adapt to salt-saturated conditions. Here, we review the current knowledge of the diversity, genomics, metabolisms and ecology of prokaryotes in DHABs.


Hypoxia-induced changes in pulmonary and systemic vascular resistance: where is the O2 sensor?
Pulmonary arteries (PA) constrict in response to alveolar hypoxia, whereas systemic arteries (SA) undergo dilation. These physiological responses reflect the need to improve gas exchange in the lung, and to enhance the delivery of blood to hypoxic systemic tissues. An important unresolved question relates to the underlying mechanism by which the vascular cells detect a decrease in oxygen tension and translate that into a signal that triggers the functional response. A growing body of work implicates the mitochondria, which appear to function as O2 sensors by initiating a redox-signaling pathway that leads to the activation of downstream effectors that regulate vascular tone. However, the direction of this redox signal has been the subject of controversy. Part of the problem has been the lack of appropriate tools to assess redox signaling in live cells. Recent advancements in the development of redox sensors have led to studies that help to clarify the nature of the hypoxia-induced redox signaling by reactive oxygen species (ROS). Moreover, these studies provide valuable insight regarding the basis for discrepancies in earlier studies of the hypoxia-induced mechanism of redox signaling. Based on recent work, it appears that the O2 sensing mechanism in both the PA and SA are identical, that mitochondria function as the site of O2 sensing, and that increased ROS release from these organelles leads to the activation of cell-specific, downstream vascular responses.


Behaviour of microtubules and actin filaments in living Drosophila embryos.
We describe the preparation of novel fluorescent derivatives of rabbit muscle actin and bovine tubulin, and the use of these derivatives to study the behaviour of actin filaments and microtubules in living Drosophila embryos, in which the nuclei divide at intervals of 8 to 21 min. The fluorescently labelled proteins appear to function normally in vitro and in vivo, and they allow continuous observation of the cytoskeleton in living embryos without perturbing development. By coinjecting labelled actin and tubulin into the early syncytial embryo, the spatial relationships between the distinct filament networks that they form can be followed second by second. The dynamic rearrangements of actin filaments and microtubules observed confirms and extends results obtained from previous studies, in which fixation techniques and specific staining were used to visualize the cytoskeleton in the Drosophila embryo. However, no tested fixation method produces an exact representation of the in vivo microtubule distribution.


Inhibiting PDE7A Enhances the Protective Effects of Neural Stem Cells on Neurodegeneration and Memory Deficits in Sevoflurane-Exposed Mice.
Sevoflurane is widely used in general anesthesia, especially for children. However, prolonged exposure to sevoflurane is reported to be associated with adverse effects on the development of brain in infant monkey. Neural stem cells (NSCs), with potent proliferation, differentiation, and renewing ability, provide an encouraging tool for basic research and clinical therapies for neurodegenerative diseases. We aim to explore the functional effects of injecting NSCs with phosphodiesterase 7A (PDE7A) knock-down in infant mice exposed to sevoflurane. The effects of PDE7A in NSCs proliferation and differentiation were determined by cell counting kit-8 (CCK-8) assay and differentiation-related gene expression assay, respectively. The effects of NSCs with modified PDE7A on mice's long-term memory and learning ability were assessed by behavioral assays. Our data demonstrated that depleting PDE7A promoted, whereas forcing PDE7A suppressed the activation of cAMP/cAMP-response element binding protein (CREB) signaling as well as cell proliferation and neuronal differentiation of NSCs. Inhibition of PDE7A in NSCs exhibited profound improved effects on long-term memory and learning ability of mice exposed to sevoflurane. Our results for the first time show that knock-down of PDE7A improves the neurogenesis of NSCs in vitro and in vivo, and is beneficial for alleviating sevoflurane-induced brain damage in infant mice.


Ketoacids? Good medicine?
D-beta-hydroxybutyrate, the principal "ketone" body in starving man, displaces glucose as the predominating fuel for brain, decreasing the need for glucose synthesis in liver (and kidney) and accordingly spares its precursor, muscle-derived amino acids. Thus normal 70 kg. man survives 2-3 months of starvation instead of several weeks, and obese man many months to over a year. Without this metabolic adaptation, H. sapiens could not have evolved such a large brain. Recent studies have shown that D-beta-hydroxybutyrate, the principal "ketone", is not just a fuel, but a "superfuel" more efficiently producing ATP energy than glucose or fatty acid. In a perfused rat heart preparation, it increased contractility and decreased oxygen consumption. It has also protected neuronal cells in tissue culture against exposure to toxins associated with Alzheimer's or Parkinson's. In a rodent model it decreased the death of lung cells induced by hemorrhagic shock. Also, mice exposed to hypoxia survived longer. These and other data suggest a potential use of beta-hydroxybutyrate in a number of medical and non-medical conditions where oxygen supply or substrate utilization may be limited. Efforts are underway to prepare esters of beta-hydroxybutyrate which can be taken orally or parenterally to study its potential therapeutic applications.


ZGLP1 is a determinant for the oogenic fate in mice.
Sex determination of germ cells is vital to creating the sexual dichotomy of germ cell development, thereby ensuring sexual reproduction. However, the underlying mechanisms remain unclear. Here, we show that ZGLP1, a conserved transcriptional regulator with GATA-like zinc fingers, determines the oogenic fate in mice. ZGLP1 acts downstream of bone morphogenetic protein, but not retinoic acid (RA), and is essential for the oogenic program and meiotic entry. ZGLP1 overexpression induces differentiation of in vitro primordial germ cell-like cells (PGCLCs) into fetal oocytes by activating the oogenic programs repressed by Polycomb activities, whereas RA signaling contributes to oogenic program maturation and PGC program repression. Our findings elucidate the mechanism for mammalian oogenic fate determination, providing a foundation for promoting in vitro gametogenesis and reproductive medicine.


Improving the 210Pb-chronology of Pb deposition in peat cores from Chao de Lamoso (NW Spain).
The natural radionuclide (210)Pb is commonly used to establish accurate and precise chronologies for the recent (past 100-150 years) layers of peat deposits. The most widely used (210)Pb-dating model, Constant Rate of Supply (CRS), was applied using data from three peat cores from Chao de Lamoso, an ombrotrophic mire in Galicia (NW Spain). On the basis of the CRS-chronologies, maximum Pb concentrations and enrichment factors (EFs) occurred in the 1960s and late 1970s, consistent with the historical use of Pb. However, maximum Pb fluxes were dated in the 1940s and the late 1960s, 10 to 20 years earlier. Principal component analysis (PCA) showed that, although the (210)Pb distribution was mainly (74%) controlled by radioactive decay, about 20% of the (210)Pb flux variability was associated with atmospheric metal pollution, suggesting an extra (210)Pb supply source and thus invalidating the main assumption of the CRS model. When the CRS-ages were recalculated after correcting for the extra input from the (210)Pb inventory of the uppermost peat layers of each core, Pb flux variations were consistent with the historical atmospheric Pb deposition. Our results not only show the robustness of the CRS model to establish accurate chronologies of recent peat deposits but also provide evidence that there are confounding factors that might influence the calculation of reliable peat accumulation rates (and thus also element accumulation rates/fluxes). This study emphasizes the need to verify the hypotheses of (210)Pb-dating models and the usefulness of a full geochemical interpretation of peat bog records.


Effect of banana powder (Musa sapientum var. paradisiaca) on gastric mucosal shedding.
Banana pulp powder (Musa sapientum Linn. var. paradisiaca) was studied for its effects on gastric mucosal resistance. Banana-treated (0.5 g/kg orally, twice daily for 3 days) rats of either sex showed: (i) a significant increase in the [3H]thymidine incorporation into mucosal cell DNA; (ii) a significant increase in the total carbohydrate (sum of total hexoses, hexosamine, fucose and sialic acid) content of gastric mucosa; (iii) a significant decrease in gastric juice DNA and protein; (iv) a significant increase in the total carbohydrates and carbohydrate/protein ratio of gastric juice. Aspirin treatment to rats caused similar effects as banana on the [3H]thymidine incorporation into mucosal cell DNA but showed opposite effects on the other parameters. These results suggest that banana treatment increased and aspirin decreased the gastric mucosal resistance as evidenced by a respective decrease and increase in gastric juice DNA, the latter serving as an index of the rate of mucosal shedding. Increased cellular mucus may be the factor for increased mucosal resistance. The results of the present study tend to confirm that plantain banana powder strengthens mucosal resistance and promotes the healing of ulcers.


Non-cirrhotic portal hypertension in HIV mono-infected patients.
Unexplained liver injury including fibrosis and portal hypertension has rarely been reported among patients with HIV in the absence of co-infection with hepatitis B (HBV) or hepatitis C (HCV). We describe a series of HIV mono-infected patients with evidence of non-cirrhotic portal hypertension. HIV-infected patients with evidence of portal hypertension who were anti-HBV and anti-HCV negative and HBV and HCV RNA polymerase chain reaction (PCR) negative were identified from patients managed by the Victorian statewide HIV referral service located at The Alfred Hospital, Melbourne. Portal hypertension was defined as either radiological or endoscopic evidence of varices, portal vein flow obstruction, or elevated hepatic venous pressure gradient (HPVG). Five patients were found to have portal hypertension. These patients were male, aged 41 to 65 years, with known duration of HIV infection between 11 to 25 years. All had been treated with antiretroviral therapy, including didanosine. Tests for metabolic, autoimmune, and hereditary causes of liver disease failed to establish an etiology for the liver injury. All had radiological or endoscopic findings of varices, and four patients had radiological features of portal vein obstruction or flow reversal. Only one patient underwent HPVG measurement, which was elevated. Non-invasive fibrosis assessment revealed increased liver stiffness in three (out of four) patients, and no cirrhotic features were found on those who underwent liver biopsy. To our knowledge, this is the largest published series of non-cirrhotic portal hypertension in HIV mono-infected patients in Australia. Further research is needed to understand what relationship, if any, HIV or its treatments might have on liver injury over time.


Shell isolated nanoparticles for enhanced Raman spectroscopy studies in lithium-oxygen cells.
A critical and detailed assessment of using Shell Isolated Nanoparticles for Enhanced Raman Spectroscopy (SHINERS) on different electrode substrates was carried out, providing relative enhancement factors, as well as an evaluation of the distribution of shell-isolated nanoparticles upon the electrode surfaces. The chemical makeup of surface layers formed upon lithium metal electrodes and the mechanism of the oxygen reduction reaction on carbon substrates relevant to lithium-oxygen cells are studied with the employment of the SHINERS technique. SHINERS enhanced the Raman signal at these surfaces showing a predominant Li2O based layer on lithium metal in a variety of electrolytes. The formation of LiO2 and Li2O2, as well as degradation reactions forming Li2CO3, upon planar carbon electrode interfaces and upon composite carbon black electrodes were followed under potential control during the reduction of oxygen in a non-aqueous electrolyte based on dimethyl sulfoxide.


Influence of calcium-induced droplet heteroaggregation on the physicochemical properties of oppositely charged lactoferrin coated lutein droplets and whey protein isolate-coated DHA droplets.
The influence of calcium-induced droplet heteroaggregation on the formation and physicochemical stability of mixed lutein and DHA emulsions was studied. Heteroaggregation was induced by mixing oppositely charged lactoferrin (LF)-coated lutein and whey protein isolate (WPI)-coated DHA emulsions with different CaCl2 concentrations at pH 6.0. The droplet size, zeta-potential, transmission-physical stability and microstructure behavior (CLSM and Cryo-SEM) of single-protein emulsions and mixed emulsions were measured as a function of different CaCl2 concentrations. Lutein degradation and DHA oxidation by measurement of lipid hydroperoxides and thiobarbituric acid reactive substances were determined during storage. The physical stability of the mixed emulsions could be modulated by controlling CaCl2 concentrations. Microstructure behavior indicated that a mixed emulsion with 30 mM CaCl2 promoted more droplets to form a special three-dimensional network and microcluster structures. The chemical stability of the mixed lutein and DHA emulsions was obviously enhanced by the addition of 30 mM CaCl2. The decreased surface areas of the DHA and lutein droplets and the physical barrier of the network of heteroaggregates against transition metals and free radicals could mainly explain the improvement in chemical stability. Calcium-induced droplet aggregation may be useful for creating specific food structures that lead to desirable physicochemical properties of multiple functional components.


Automated de novo phasing and model building of coiled-coil proteins.
Models generated by de novo structure prediction can be very useful starting points for molecular replacement for systems where suitable structural homologues cannot be readily identified. Protein-protein complexes and de novo-designed proteins are examples of systems that can be challenging to phase. In this study, the potential of de novo models of protein complexes for use as starting points for molecular replacement is investigated. The approach is demonstrated using homomeric coiled-coil proteins, which are excellent model systems for oligomeric systems. Despite the stereotypical fold of coiled coils, initial phase estimation can be difficult and many structures have to be solved with experimental phasing. A method was developed for automatic structure determination of homomeric coiled coils from X-ray diffraction data. In a benchmark set of 24 coiled coils, ranging from dimers to pentamers with resolutions down to 2.5 Å, 22 systems were automatically solved, 11 of which had previously been solved by experimental phasing. The generated models contained 71-103% of the residues present in the deposited structures, had the correct sequence and had free R values that deviated on average by 0.01 from those of the respective reference structures. The electron-density maps were of sufficient quality that only minor manual editing was necessary to produce final structures. The method, named CCsolve, combines methods for de novo structure prediction, initial phase estimation and automated model building into one pipeline. CCsolve is robust against errors in the initial models and can readily be modified to make use of alternative crystallographic software. The results demonstrate the feasibility of de novo phasing of protein-protein complexes, an approach that could also be employed for other small systems beyond coiled coils.


Enhanced flavor-nutrient conditioning in obese rats on a high-fat, high-carbohydrate choice diet.
Through flavor-nutrient conditioning rats learn to prefer and increase their intake of flavors paired with rewarding, postingestive nutritional consequences. Since obesity is linked to altered experience of food reward and to perturbations of nutrient sensing, we investigated flavor-nutrient learning in rats made obese using a high fat/high carbohydrate (HFHC) choice model of diet-induced obesity (ad libitum lard and maltodextrin solution plus standard rodent chow). Forty rats were maintained on HFHC to induce substantial weight gain, and 20 were maintained on chow only (CON). Among HFHC rats, individual differences in propensity to weight gain were studied by comparing those with the highest proportional weight gain (obesity prone, OP) to those with the lowest (obesity resistant, OR). Sensitivity to postingestive food reward was tested in a flavor-nutrient conditioning protocol. To measure initial, within-meal stimulation of flavor acceptance by post-oral nutrient sensing, first, in sessions 1-3, baseline licking was measured while rats consumed grape- or cherry-flavored saccharin accompanied by intragastric (IG) water infusion. Then, in the next three test sessions they received the opposite flavor paired with 5 ml of IG 12% glucose. Finally, after additional sessions alternating between the two flavor-infusion contingencies, preference was measured in a two-bottle choice between the flavors without IG infusions. HFHC-OP rats showed stronger initial enhancement of intake in the first glucose infusion sessions than CON or HFHC-OR rats. OP rats also most strongly preferred the glucose-paired flavor in the two-bottle choice. These differences between OP versus OR and CON rats suggest that obesity is linked to responsiveness to postoral nutrient reward, consistent with the view that flavor-nutrient learning perpetuates overeating in obesity.


Huntington's disease genotype suppresses global manganese-responsive processes in pre-manifest and manifest YAC128 mice.
Manganese (Mn) is an essential micronutrient required for the proper function of several enzymes. Accumulating evidence demonstrates a selective decrease of bioavailable Mn in vulnerable cell types of Huntington's Disease (HD), an inherited progressive neurodegenerative disorder with no cure. Amelioration of underlying pathophysiology, such as alterations in Mn-dependent biology, may be therapeutic. We therefore sought to investigate global Mn-dependent and Mn-responsive biology following various Mn exposures in a mouse model of HD. YAC128 and wildtype (WT) littermate control mice received one of three different Mn exposure paradigms by subcutaneous injection of 50 mg kg-1 MnCl2·4(H2O) across two distinct HD disease stages. "Pre-manifest" (12-week old mice) mice received either a single (1 injection) or week-long (3 injections) exposure of Mn or vehicle (H2O) and were sacrificed at the pre-manifest stage. "Manifest" (32-week old) mice were sacrificed following either a week-long Mn or vehicle exposure during the manifest stage, or a 20-week-long chronic (2× weekly injections) exposure that began in the pre-manifest stage. Tissue Mn, mRNA, protein, and metabolites were measured in the striatum, the brain region most sensitive to neurodegeneration in HD. Across all Mn exposure paradigms, pre-manifest YAC128 mice exhibited a suppressed response to transcriptional and protein changes and manifest YAC128 mice showed a suppressed metabolic response, despite equivalent elevations in whole striatal Mn. We conclude that YAC128 mice respond differentially to Mn compared to WT as measured by global transcriptional, translational, and metabolomic changes, suggesting an impairment in Mn homeostasis across two different disease stages in YAC128 mice.


COVID-19-Related Hospitalization Rates and Severe Outcomes Among Veterans From 5 Veterans Affairs Medical Centers: Hospital-Based Surveillance Study.
COVID-19 has disproportionately affected older adults and certain racial and ethnic groups in the United States. Data quantifying the disease burden, as well as describing clinical outcomes during hospitalization among these groups, are needed. We aimed to describe interim COVID-19 hospitalization rates and severe clinical outcomes by age group and race and ethnicity among US veterans by using a multisite surveillance network. We implemented a multisite COVID-19 surveillance platform in 5 Veterans Affairs Medical Centers located in Atlanta, Bronx, Houston, Palo Alto, and Los Angeles, collectively serving more than 396,000 patients annually. From February 27 to July 17, 2020, we actively identified inpatient cases with COVID-19 by screening admitted patients and reviewing their laboratory test results. We then manually abstracted the patients' medical charts for demographics, underlying medical conditions, and clinical outcomes. Furthermore, we calculated hospitalization incidence and incidence rate ratios, as well as relative risk for invasive mechanical ventilation, intensive care unit admission, and case fatality rate after adjusting for age, race and ethnicity, and underlying medical conditions. We identified 621 laboratory-confirmed, hospitalized COVID-19 cases. The median age of the patients was 70 years, with 65.7% (408/621) aged ≥65 years and 94% (584/621) male. Most COVID-19 diagnoses were among non-Hispanic Black (325/621, 52.3%) veterans, followed by non-Hispanic White (153/621, 24.6%) and Hispanic or Latino (112/621, 18%) veterans. Hospitalization rates were the highest among veterans who were ≥85 years old, Hispanic or Latino, and non-Hispanic Black (430, 317, and 298 per 100,000, respectively). Veterans aged ≥85 years had a 14-fold increased rate of hospitalization compared with those aged 18-29 years (95% CI: 5.7-34.6), whereas Hispanic or Latino and Black veterans had a 4.6- and 4.2-fold increased rate of hospitalization, respectively, compared with non-Hispanic White veterans (95% CI: 3.6-5.9). Overall, 11.6% (72/621) of the patients required invasive mechanical ventilation, 26.6% (165/621) were admitted to the intensive care unit, and 16.9% (105/621) died in the hospital. The adjusted relative risk for invasive mechanical ventilation and admission to the intensive care unit did not differ by age group or race and ethnicity, but veterans aged ≥65 years had a 4.5-fold increased risk of death while hospitalized with COVID-19 compared with those aged <65 years (95% CI: 2.4-8.6). COVID-19 surveillance at the 5 Veterans Affairs Medical Centers across the United States demonstrated higher hospitalization rates and severe outcomes among older veterans, as well as higher hospitalization rates among Hispanic or Latino and non-Hispanic Black veterans than among non-Hispanic White veterans. These findings highlight the need for targeted prevention and timely treatment for veterans, with special attention to older aged, Hispanic or Latino, and non-Hispanic Black veterans.


Complex interplay between intrinsic and extrinsic drivers of long-term survival trends in southern elephant seals.
Determining the relative contribution of intrinsic and extrinsic factors to fluctuations in population size, trends and demographic composition is analytically complex. It is often only possible to examine the combined effects of these factors through measurements made over long periods, spanning an array of population densities or levels of food availability. Using age-structured mark-recapture models and datasets spanning five decades (1950-1999), and two periods of differing relative population density, we estimated age-specific probabilities of survival and examined the combined effects of population density and environmental conditions on juvenile survival of southern elephant seals at Macquarie Island. First-year survival decreased with density during the period of highest population size, and survival increased during years when the Southern Oscillation Index (SOI) anomaly (deviation from a 50-year mean) during the mother's previous foraging trip to sea was positive (i.e., El Niño). However, when environmental stochasticity and density were considered together, the effect of density on first-year survival effectively disappeared. Ignoring density effects also leads to models placing too much emphasis on the environmental conditions prevailing during the naïve pup's first year at sea. Our analyses revealed that both the state of the environment and population density combine to modify juvenile survival, but that the degree to which these processes contributed to the variation observed was interactive and complex. This underlines the importance of evaluating the relative contribution of both the intrinsic and extrinsic factors that regulate animal populations because false conclusions regarding the importance of population regulation may be reached if they are examined in isolation.


Cellular distribution of transcripts for tissue inhibitor of metalloproteinases 1 and 2 in human hepatocellular carcinomas.
The cellular distribution of tissue inhibitor of metalloproteinases (TIMP)-1, and TIMP-2 was studied by using in situ hybridization in surgically removed human hepatocellular carcinomas (HCCs) and cholangiocellular carcinomas (CCCs). The purpose of this study was to characterize the protein involvement of TIMPs in the development of HCCs and CCCs. All HCCs and CCCs expressed TIMPs. The distribution of transcripts for TIMPs in the tumors was mostly homogeneous. Expression of TIMP in cancer cells was more intense than that in the surrounding noncancerous liver (either, cirrhosis, chronic hepatitis, or normal), and expression of TIMP-1 was stronger than that of TIMP-2. Expression of TIMPs varied among HCC nodules, but there was no obvious association between the expression level of TIMPs and differentiation stages or invasiveness of the HCCs. Transcripts for TIMPs were clearly demonstrated in the metastatic HCC nodules in the lung. Expression of TIMP-1 CCC was strong, and small nodules of CCC were recognized in the liver. Immunohistochemical study for TIMP-1 revealed a consistent staining of the TIMP protein with the transcripts. In the peritumoral histologically normal liver, which was not infected with either hepatitis B or C virus, expression of TIMP-1 was found in various cell types, but that of TIMP-2 was weak. Expression of TIMP-1 in hepatocytes revealed clear zonal distribution. These results suggest that TIMPs may act on modulating the matrix/tumor interaction and may play an important role in growth and invasion of HCCs and CCCs. Expression of TIMP-1 can be a marker of HCC metastasis to the lung, and also that of the extent of CCC invasion. Furthermore, the consistent expression of TIMPs in many cell types of the noncancerous liver suggests some unknown functional role that must be clarified.


Intratumoral diversity of telomere length in individual neuroblastoma tumors.
The purpose of the work was to investigate telomere length (TL) and mechanisms involved in TL maintenance in individual neuroblastoma (NB) tumors. Primary NB tumors from 102 patients, ninety Italian and twelve Spanish, diagnosed from 2000 to 2008 were studied. TL was investigated by quantitative fluorescence in situ hybridization (IQ-FISH) that allows to analyze individual cells in paraffin-embedded tissues. Fluorescence intensity of chromosome 2 centromere was used as internal control to normalize TL values to ploidy. Human telomerase reverse transcriptase (hTERT) expression was detected by immunofluorescence in 99/102 NB specimens.The main findings are the following: 1) two intratumoral subpopulations of cancer cells displaying telomeres of different length were identified in 32/102 tumors belonging to all stages. 2) hTERT expression was detected in 99/102 tumors, of which 31 displayed high expression and 68 low expression. Alternative lengthening of telomeres (ALT)-mechanism was present in 60/102 tumors, 20 of which showed high hTERT expression. Neither ALT-mechanism nor hTERT expression correlated with heterogeneous TL. 3) High hTERT expression and ALT positivity were associated with significantly reduced Overall Survival. 4) High hTERT expression predicted relapse irrespective of patient age. Intratumoral diversity in TL represents a novel feature in NB.In conclusion, diversity of TL in individual NB tumors was strongly associated with disease progression and death, suggesting that these findings are of translational relevance. The combination of high hTERT expression and ALT positivity may represent a novel biomarker of poor prognosis that deserves further investigation.


Prefrontal cortical activity predicts the occurrence of nonlocal hippocampal representations during spatial navigation.
The receptive field of a neuron describes the regions of a stimulus space where the neuron is consistently active. Sparse spiking outside of the receptive field is often considered to be noise, rather than a reflection of information processing. Whether this characterization is accurate remains unclear. We therefore contrasted the sparse, temporally isolated spiking of hippocampal CA1 place cells to the consistent, temporally adjacent spiking seen within their spatial receptive fields ("place fields"). We found that isolated spikes, which occur during locomotion, are strongly phase coupled to hippocampal theta oscillations and transiently express coherent nonlocal spatial representations. Further, prefrontal cortical activity is coordinated with and can predict the occurrence of future isolated spiking events. Rather than local noise within the hippocampus, sparse, isolated place cell spiking reflects a coordinated cortical-hippocampal process consistent with the generation of nonlocal scenario representations during active navigation.


Does hybridization of endophytic symbionts in a native grass increase fitness in resource-limited environments?
Hybridization is common among plants, animals and microbes. However, the ecological consequences of hybridization for microbes are far less understood than for plants and animals. For symbiotic Epichloë fungi, hybridization is widespread and may augment the well-known benefits of the endophytes to their grass hosts, especially in stressful environments. We tested the hybrid fitness hypothesis (HFH) that hybrid endophytes enhance fitness in stressful environments relative to non-hybrid endophytes. In a long-term field experiment, we monitored growth and reproduction of hybrid-infected (H+), non-hybrid infected (NH+), naturally endophyte free (E-) plants and those plants from which the endophyte had been experimentally removed (H- and NH-) in resource-rich and resource-poor environments. Infection by both endophyte species enhanced growth and reproduction. H+ plants outperformed NH+ plants in terms of growth by the end of the experiment, supporting HFH. However, H+ plants only outperformed NH+ plants in the resource-rich treatment, contrary to HFH. Plant genotypes associated with each endophyte species had strong effects on growth and reproduction. Our results provide some support the HFH hypothesis but not based upon adaptation to stressful environments. Our results reinforce the notion of a complex interplay between endophyte and plant genotype and environmental factors that determine fitness of the symbiotum.


Expressions for the nonlinear transmission performance of multi-mode optical fiber.
We develop an analytical theory which allows us to identify the information spectral density limits of multimode optical fiber transmission systems. Our approach takes into account the Kerr-effect induced interactions of the propagating spatial modes and derives closed-form expressions for the spectral density of the corresponding nonlinear distortion. Experimental characterization results have confirmed the accuracy of the proposed models. Application of our theory in different FMF transmission scenarios has predicted a ~10% variation in total system throughput due to changes associated with inter-mode nonlinear interactions, in agreement with an observed 3dB increase in nonlinear noise power spectral density for a graded index four LP mode fiber.


Guatemalan school food environment: impact on schoolchildren's risk of both undernutrition and overweight/obesity.
Guatemala suffers the double burden of malnutrition with high rates of stunting alongside increasing childhood overweight/obesity. This study examines the school food environment (SFE) at low-income Guatemalan elementary schools and discusses its potential impact on undernutrition and overweight/obesity. From July through October 2013, direct observations, in-depth interviews with school principals (n = 4) and food kiosk vendors (n = 4, 2 interviews each) and also focus groups (FGs) with children (n = 48, 8 FGs) were conducted. The SFE comprises food from school food kiosks (casetas); food from home or purchased in the street; and food provided by the school (refacción). School casetas, street vendors and children's parents largely provide sandwiches, calorie-rich snacks and sugar-sweetened beverages. Refacción typically serves energy dense atol, a traditional beverage. The current school food program (refacción), the overall SFE and the roles/opinions of vendors and principals reveal persistent anxiety concerning undernutrition and insufficient concern for overweight/obesity. Predominant concern for elementary schoolchildren remains focused on undernutrition. However, by the time children reach elementary school (ages 6-12+), food environments should encourage dietary behaviors to prevent childhood overweight/obesity.


Calcium imaging of odor-evoked responses in the Drosophila antennal lobe.
The antennal lobe is the primary olfactory center in the insect brain and represents the anatomical and functional equivalent of the vertebrate olfactory bulb. Olfactory information in the external world is transmitted to the antennal lobe by olfactory sensory neurons (OSNs), which segregate to distinct regions of neuropil called glomeruli according to the specific olfactory receptor they express. Here, OSN axons synapse with both local interneurons (LNs), whose processes can innervate many different glomeruli, and projection neurons (PNs), which convey olfactory information to higher olfactory brain regions. Optical imaging of the activity of OSNs, LNs and PNs in the antennal lobe - traditionally using synthetic calcium indicators (e.g. calcium green, FURA-2) or voltage-sensitive dyes (e.g. RH414) - has long been an important technique to understand how olfactory stimuli are represented as spatial and temporal patterns of glomerular activity in many species of insects. Development of genetically-encoded neural activity reporters, such as the fluorescent calcium indicators G-CaMP and Cameleon, the bioluminescent calcium indicator GFP-aequorin, or a reporter of synaptic transmission, synapto-pHluorin has made the olfactory system of the fruitfly, Drosophila melanogaster, particularly accessible to neurophysiological imaging, complementing its comprehensively-described molecular, electrophysiological and neuroanatomical properties. These reporters can be selectively expressed via binary transcriptional control systems (e.g. GAL4/UAS, LexA/LexAop, Q system) in defined populations of neurons within the olfactory circuitry to dissect with high spatial and temporal resolution how odor-evoked neural activity is represented, modulated and transformed. Here we describe the preparation and analysis methods to measure odor-evoked responses in the Drosophila antennal lobe using G-CaMP. The animal preparation is minimally invasive and can be adapted to imaging using wide-field fluorescence, confocal and two-photon microscopes.


Proteomics application exercise of the Swiss Proteomics Society: report of the SPS'02 session.
After the success of the mass spectrometry (MS) round table that was held at the first Swiss Proteomics Society congress (SPS'01) in Geneva, the SPS has organized a proteomics application exercise and allocated a full session at the SPS'02 congress. The main objective was to encourage the exchange of expertise in protein identification, with a focus on the use of mass spectrometry, and to create a bridge between the users' questions and the instrument providers' solutions. Two samples were sent to fifteen interested labs, including academic groups and MS hardware providers. Participants were asked to identify and partially characterize the samples. They consisted of a complex mixture of peptide/proteins (sample A) and an almost pure recombinant peptide carrying post-translational modifications (sample B). Sample A was an extract of snake venom from the species Bothrops jararaca. Sample B was a recombinant and modified peptide derived from the shrimp Penaeus vannamei penaeidin 3a. The eight labs that returned results reported the use of a wide range of MS instrumentation and techniques. They mentioned a variety of time and manpower allocations. The origin of sample A was generally identified together with a number of database protein entries. The difficulty of the sample identification lay in the incomplete knowledge of the Bothrops species genome sequence and is discussed. Sample B was generally and correctly identified as penaeidin. However, only one group reported the full primary structure. Interestingly, the approaches were again varied and are discussed in the text.


Engineering the rRNA decoding site of eukaryotic cytosolic ribosomes in bacteria.
Structural and genetic studies on prokaryotic ribosomes have provided important insights into fundamental aspects of protein synthesis and translational control and its interaction with ribosomal drugs. Comparable mechanistic studies in eukaryotes are mainly hampered by the absence of both high-resolution crystal structures and efficient genetic models. To study the interaction of aminoglycoside antibiotics with selected eukaryotic ribosomes, we replaced the bacterial drug binding site in 16S rRNA with its eukaryotic counterpart, resulting in bacterial hybrid ribosomes with a fully functional eukaryotic rRNA decoding site. Cell-free translation assays demonstrated that hybrid ribosomes carrying the rRNA decoding site of higher eukaryotes show pronounced resistance to aminoglycoside antibiotics, equivalent to that of rabbit reticulocyte ribosomes, while the decoding sites of parasitic protozoa show distinctive drug susceptibility. Our findings suggest that phylogenetically variable components of the ribosome, other than the rRNA-binding site, do not affect aminoglycoside susceptibility of the protein-synthesis machinery. The activities of the hybrid ribosomes indicate that helix 44 of the rRNA decoding site behaves as an autonomous domain, which can be exchanged between ribosomes of different phylogenetic domains for study of function.


Performance evaluation of the microPET P4: a PET system dedicated to animal imaging.
The microPET Primate 4-ring system (P4) is an animal PET tomograph with a 7.8 cm axial extent, a 19 cm diameter transaxial field of view (FOV) and a 22 cm animal port. The system is composed of 168 detector modules, each with an 8 x 8 array of 2.2 x 2.2 x 10 mm3 lutetium oxyorthosilicate crystals, arranged as 32 crystal rings 26 cm in diameter. The detector crystals are coupled to a Hamamatsu R5900-C8 PS-PMT via a 10 cm long optical fibre bundle. The detectors have a timing resolution of 3.2 ns, an average energy resolution of 26%, and an average intrinsic spatial resolution of 1.75 mm. The system operates in 3D mode without inter-plane septa, acquiring data in list mode. The reconstructed image spatial resolution ranges from 1.8 mm at the centre to 3 mm at 4 cm radial offset. The tomograph has a peak system sensitivity of 2.25% at the centre of the FOV with a 250-750 keV energy window. The noise equivalent count rate peaks at 100-290 kcps for representative object sizes. Images from two phantoms and three different types of laboratory animal demonstrate the advantage of the P4 system over the original prototype microPET. including its threefold improvement in sensitivity and a large axial FOV sufficient to image an entire mouse in a single bed position.


The use of fresh-frozen plasma in England: high levels of inappropriate use in adults and children.
Fresh-frozen plasma (FFP) is given to patients across a range of clinical settings, frequently in association with abnormalities of standard coagulation tests. A UK-wide study of FFP transfusion practice was undertaken to characterize the current patterns of administration and to evaluate the contribution of pretransfusion coagulation tests. A total of 4969 FFP transfusions given to patients in 190 hospitals were analyzed, of which 93.3% were in adults and 6.7% in children or infants. FFP transfusions to adults were given most frequently in intensive-treatment or high-dependency units (32%), in operating rooms or recovery (23%), or on medical wards (22%). In adult patients 43% of all FFP transfusions were given in the absence of documented bleeding, as prophylaxis for abnormal coagulation tests or before procedures or surgery. There was wide variation in international normalized ratio (INR) or prothrombin times before FFP administration; in 30.9% of patients where the main reason for transfusion was prophylactic in the absence of bleeding the INR was 1.5 or less. Changes in standard coagulation results after FFP administration were generally very small for adults and children. This study raises important questions about the clinical benefit of much of current FFP usage. It highlights the pressing need for better studies to inform and evaluate quantitative data for the effect of plasma on standard coagulation tests.


Correction for Eddy Current-Induced Echo-Shifting Effect in Partial-Fourier Diffusion Tensor Imaging.
In most diffusion tensor imaging (DTI) studies, images are acquired with either a partial-Fourier or a parallel partial-Fourier echo-planar imaging (EPI) sequence, in order to shorten the echo time and increase the signal-to-noise ratio (SNR). However, eddy currents induced by the diffusion-sensitizing gradients can often lead to a shift of the echo in k-space, resulting in three distinct types of artifacts in partial-Fourier DTI. Here, we present an improved DTI acquisition and reconstruction scheme, capable of generating high-quality and high-SNR DTI data without eddy current-induced artifacts. This new scheme consists of three components, respectively, addressing the three distinct types of artifacts. First, a k-space energy-anchored DTI sequence is designed to recover eddy current-induced signal loss (i.e., Type 1 artifact). Second, a multischeme partial-Fourier reconstruction is used to eliminate artificial signal elevation (i.e., Type 2 artifact) associated with the conventional partial-Fourier reconstruction. Third, a signal intensity correction is applied to remove artificial signal modulations due to eddy current-induced erroneous T2(∗) -weighting (i.e., Type 3 artifact). These systematic improvements will greatly increase the consistency and accuracy of DTI measurements, expanding the utility of DTI in translational applications where quantitative robustness is much needed.


The plover neurotranscriptome assembly: transcriptomic analysis in an ecological model species without a reference genome.
We assembled a de novo transcriptome of short-read Illumina RNA-Seq data generated from telencephalon and diencephalon tissue samples from the Kentish plover, Charadrius alexandrinus. This is a species of considerable interest in behavioural ecology for its highly variable mating system and parental behaviour, but it lacks genomic resources and is evolutionarily distant from the few available avian draft genome sequences. We assembled and identified over 21,000 transcript contigs with significant expression in our samples, showing high homology to exonic sequences in avian draft genomes. From these, we identified >31,000 high-quality SNPs and > 2500 simple sequence repeats (SSRs). We also analysed expression patterns in our data to identify potential candidate genes related to differences in male and female behaviour, identifying over 200 nonoverlapping putative autosomal transcripts that show significant expression differences between males and females. Gene ontology analysis revealed that female-biased transcripts were significantly enriched for cerebral functions related to learning, cognition and memory, and male-biased transcripts were mostly enriched for terms related to neural function such as neuron projection and synapses. This data set provides one of the first de novo transcriptome assemblies from non-normalized short-read next-generation data and outlines an effective strategy for measuring sequence and expression variability simultaneously without the aid of a reference genome.


Determinants of mood in general practitioners.
A pilot study was conducted in which 44 general practitioners completed cognitive behavioural self monitoring diaries. Hourly changes in emotional state were recorded together with associated circumstances. Lowering of mood was associated mainly with "hassle" at work, pressure of time, and domestic dissatisfaction. Improvement in mood was associated with domestic happiness and satisfaction at working efficiently and to time. Mood was significantly lower when the doctor was on call. Women doctors were more prone to mood changes associated with domestic matters. Responses to a questionnaire suggested that the doctors preferred traditional clinical medicine to problems of a social or psychological origin. Managerial skills would help alleviate several of the problems identified in this study and should be more prominent in the training that all doctors receive.


Increasing available NADH supply during succinic acid production by Corynebacterium glutamicum.
A critical factor in the biotechnological production of succinic acid with Corynebacterium glutamicum is the sufficient supply of NADH. It is conceivable that cofactor availability and the proportion of cofactor in the active form may play an important role in dictating the succinic acid yield. PntAB genes from Escherichia coli can directly catalyze the reversible hydride transfer and adjust the dynamic balance between NADP(H) and NAD(H). Hence, we studied the physiological effect of coenzyme systems by expressing the membrane-bound transhydrogenase pntAB genes. We have shown experimentally that the pntAB genes could function as an alternative source of NADH. In an anaerobic fermentation with C. glutamicum NC-3-pntAB, a 16% higher succinic acid yield and a 57% higher production from glucose were obtained by pntAB expression. Moreover, the formation of by-products was significantly decreased. The concomitant increase in the consumption of intracellular NADPH from 0.6 to 1.2 mmol/g CDW and the increased NADH/NAD(+) ratio resulted from introduction of pntAB, suggesting that the membrane-bound transhydrogenase converted excess NADPH to NADH for succinic acid production. Finally, we explored whether the transhydrogenase had different effects on the succinic acid formation on different carbon sources. The succinic acid yield was increased in the presence of pntAB by 16% on glucose, 7% on sucrose, and without large influence on fructose and xylose. The results of this study demonstrated that the effectiveness of cofactor manipulation could be a promising strategy applied in metabolic engineering.


Empathy: gender effects in brain and behavior.
Evidence suggests that there are differences in the capacity for empathy between males and females. However, how deep do these differences go? Stereotypically, females are portrayed as more nurturing and empathetic, while males are portrayed as less emotional and more cognitive. Some authors suggest that observed gender differences might be largely due to cultural expectations about gender roles. However, empathy has both evolutionary and developmental precursors, and can be studied using implicit measures, aspects that can help elucidate the respective roles of culture and biology. This article reviews evidence from ethology, social psychology, economics, and neuroscience to show that there are fundamental differences in implicit measures of empathy, with parallels in development and evolution. Studies in nonhuman animals and younger human populations (infants/children) offer converging evidence that sex differences in empathy have phylogenetic and ontogenetic roots in biology and are not merely cultural byproducts driven by socialization. We review how these differences may have arisen in response to males' and females' different roles throughout evolution. Examinations of the neurobiological underpinnings of empathy reveal important quantitative gender differences in the basic networks involved in affective and cognitive forms of empathy, as well as a qualitative divergence between the sexes in how emotional information is integrated to support decision making processes. Finally, the study of gender differences in empathy can be improved by designing studies with greater statistical power and considering variables implicit in gender (e.g., sexual preference, prenatal hormone exposure). These improvements may also help uncover the nature of neurodevelopmental and psychiatric disorders in which one sex is more vulnerable to compromised social competence associated with impaired empathy.


A systematic review of interventions to improve outcomes for elders discharged from the emergency department.
To evaluate the evidence for interventions designed to improve outcomes for elders discharged from the emergency department (ED). The study was a systematic review of English-language articles indexed in MEDLINE and CINAHL (1966-2005) with 1) key words "geriatric," "older adults," or "seniors," or 2) Medical Subject Heading (MeSH) terms "Geriatrics" or "Health Services for the Aged" AND key word "emergency," or 3) MeSH terms "Emergencies," "Emergency Service, Hospital," or "Emergency Treatment." Bibliographies of the retrieved articles were reviewed for additional references, and the authors consulted with content experts to identify relevant unpublished work. Patients of interest were community-dwelling elder patients discharged home from the ED. Data were abstracted from selected articles by the authors. Studies with interventions limited to patients with a single presentation or diagnosis (falls, delirium, etc.) or delivered only to patients who would have otherwise been hospitalized were not included. Of 669 citations, 27 studies (reported in 33 articles) met study criteria and were reviewed; six randomized controlled trials (RCTs), two nonrandomized clinical trials, and 19 observational studies or program descriptions. Three of four RCTs designed to measure functional outcomes showed a reduction in functional decline in the intervention group. The trials that resulted in functional benefits enrolled high-risk patients and included geriatric nursing assessment and home-based services as part of the intervention. Results of trials to decrease health service utilization rates following an ED visit were mixed. A significant number of programs to improve outcomes for elders discharged from the ED exist, but few have been systematically examined. Development of interventions to improve the care of elder patients following ED visits requires further research into system and patient-centered factors that impact health care delivery in this situation.


Brain derived neurotrophic factor gene Val66Met and -270C/T polymorphisms and personality traits predisposing to anorexia nervosa.
Polymorphisms in BDNF gene has been proposed, as susceptibility loci for stress-related psychiatric disorders. Several lines of molecular and biochemical evidence point to the role of BDNF in anorexia nervosa (AN). Personality traits may constitute the intermediate phenotypes between genes and vulnerability to AN. BDNF Val66Met and -270C/T polymorphisms were genotyped in 149 patients with AN and 100 healthy control females. Temperamental traits in all subjects were measured with Temperament and Character Inventory. First in case-control analysis, we assessed, if analyzed genotypes confer risk for AN. Next, the association of BDNF gene variants with personality dimensions in patients and control subjects was analyzed. No significant differences between patients with anorexia nervosa and controls in frequency of genotypes and alleles were observed. AN patients with Met allele showed higher Harm avoidance (Anova F=4.70; p=0.03) than Val/Val homozygotes. AN patients, who carried the T allele of BDNF -270C/T polymorphism showed higher Persistence (Anova F=4.04; p<0.05) and Harm avoidance (Anova F=7.93; p=0.006) than C/C homozygotes, however after correction for multiple testing only the latter association remained statistically significant. No significant relationship between Val/Met 66 genotype and -270C/T genotype with personality was observed in healthy females. These results may suggest, that BDNF -270 C/T polymorphism may influence the personality trait associated with higher risk of AN.


HAART is neuroprophylactic in HIV-1 infection.
To find out about the prophylactic value of antiretroviral therapy on HIV-1-associated subclinical and clinical psychomotor slowing as one marker of HIV-1-associated CNS disease. Prospective study with regular clinical and neurophysiologic examination every three months of 1482 consecutive HIV-1-seropositive and AIDS patients seen at our department till June 30, 1999. Antiretroviral therapy has a significant prophylactic value over an individual observation period of ten years with regard to the first, potentially transient manifestation of HIV-1-associated subclinical psychomotor slowing and with regard to the clinical manifestation of motor signs. However, a subgroup of patients is characterized through a second, more sustained manifestation of subclinical psychomotor slowing which cannot be prevented by any type of currently available antiretroviral therapy. These findings suggest the existence of different pathomechanisms underlying HIV-1-associated brain disease which may in part be effectively prevented, but which in part also escape all antiretroviral treatment strategies in use today.


Asymmetric global motion integration in drifting Gabor arrays.
We examined how ambiguous motion signals are integrated over space to support the unambiguous perception of global motion. The motion of a Gaussian windowed drifting sine grating (Gabor) is consistent with an infinite number of grating velocities. To extract the consistent global motion of multi-Gabor arrays, the visual system must integrate ambiguous motion signals from disparate regions of visual space. We found an interaction between spatial arrangement and global motion integration in this process. Linear arrays of variably oriented Gabor elements appeared to move more slowly, reflecting suboptimal integration, when the direction of global translation was orthogonal to the line as opposed to along it. Circular arrays of Gabor elements appeared to move more slowly when the global motion was an expansion or contraction rather than a rotation. However, there was no difference in perceived speed for densely packed annular arrays for these global motion pattern directions. We conclude that the region over which ambiguous motion is integrated is biased in the direction of global motion, and the concept of the association field, held to link like elements along a contour, needs to be extended to include global motion computation over disparate elements referencing the same global motion.


Form and function of the teleost lateral line revealed using three-dimensional imaging and computational fluid dynamics.
Fishes sense weak water motion using the lateral line. Among the thousands of described fish species, this organ may differ in size, shape and distribution of individual mechanoreceptors or lateral line canals. The reasons for this diversity remain unclear, but are very likely related to habitat preferences. To better understand the performance of the organ in natural hydrodynamic surroundings, various three-dimensional imaging datasets of the cephalic lateral line were gathered using Leuciscus idus as representative freshwater teleost. These data are employed to simulate hydrodynamic phenomena around the head and within lateral line canals. The results show that changes in canal dimensions alter the absolute stimulation amplitudes, but have little effect on the relation between bulk water flow and higher frequency signals. By contrast, depressions in the skin known as epidermal pits reduce bulk flow stimulation and increase the ratio between higher-frequency signals and the background flow stimulus.


Identification, characterization, and developmental regulation of a receptor guanylyl cyclase expressed during early stages of Drosophila development.
Membrane forms of guanylyl cyclase are single-transmembrane proteins that are activated by the binding of specific peptide ligands to their extracellular domains. In this report, we describe the identification and characterization of a Drosophila cDNA clone encoding a protein, DrGC-1, with high sequence identity to members of this family of receptor proteins. The protein contains a single, hydrophobic domain predicted to represent a transmembrane segment separating an extracellular domain with significant sequence identity (30%) to sea urchin egg peptide receptors from intracellular domains containing a protein kinase-like domain followed by a region with high sequence identity (65%) to cyclase catalytic domains found in receptor guanylyl cyclases from both vertebrates and invertebrates. In contrast to other members of this family, DrGC-1 is predicted to contain a carboxyl-terminal extension of 430 residues that has no homology to any described protein. Northern analysis indicates that DrGC-1 transcripts are present at variable levels in all stages of development. In situ hybridization demonstrates that high levels of uniformly distributed transcript are present in 0-2-h embryos. Later in embryogenesis (14-18 h), elevated levels of hybridization appear to be preferentially associated with muscle fibers.


RNAexinv: An extended inverse RNA folding from shape and physical attributes to sequences.
RNAexinv is an interactive java application that performs RNA sequence design, constrained to yield a specific RNA shape and physical attributes. It is an extended inverse RNA folding program with the rationale behind that the generated sequences should not only fold into a desired structure, but they should also exhibit favorable attributes such as thermodynamic stability and mutational robustness. RNAexinv considers not only the secondary structure in order to design sequences, but also the mutational robustness and the minimum free energy. The sequences that are generated may not fully conform with the given RNA secondary structure, but they will strictly conform with the RNA shape of the given secondary structure and thereby take into consideration the recommended values of thermodynamic stability and mutational robustness that are provided. The output consists of designed sequences that are generated by the proposed method. Selecting a sequence displays the secondary structure drawings of the target and the predicted fold of the sequence, including some basic information about the desired and achieved thermodynamic stability and mutational robustness. RNAexinv can be used successfully without prior experience, simply specifying an initial RNA secondary structure in dot-bracket notation and numerical values for the desired neutrality and minimum free energy. The package runs under LINUX operating system. Secondary structure predictions are performed using the Vienna RNA package. RNAexinv is a user friendly tool that can be used for RNA sequence design. It is especially useful in cases where a functional stem-loop structure of a natural sequence should be strictly kept in the designed sequences but a distant motif in the rest of the structure may contain one more or less nucleotide at the expense of another, as long as the global shape is preserved. This allows the insertion of physical observables as constraints. RNAexinv is available at http://www.cs.bgu.ac.il/~RNAexinv.


Practice visits as a tool in quality improvement: acceptance and feasibility.
To evaluate the feasibility and acceptance of (a) two programmes of assessment of practice management in a practice visit: mutual practice visits and feedback by peers versus visits and feedback by non-physician observers and (2) the practice visit method used in these programmes (the visit instrument to assess practice management and organisation (VIP)--a validated Dutch tool). Prospective, randomised intervention study with the two programmes, follow up after one year. General practitioners (GPs) were visited after each programme and after the revisits by non-physician observers a year later. General practices in the Netherlands in 1993 and 1994. A total of 90 GPs in 68 practices. At follow up after 1 year there were 81 GPs in 62 practices. Scores (mainly five point scales) for questions on appreciation and acceptance; after the follow up visit a year later, scores for questions on feasibility and practicality of the improved procedure and feedback report. Data of 44 mutual visits by peers were compared with data of 46 visits by non-physician observers. A visit by a non-physician observer was appreciated significantly more. After the practice visit at one year follow up, the participants reported to have appreciated the visit and the feedback report and to prefer feedback of a non-physician observer to that of a peer. Participants' reports on the procedure and the presentation of the feedback provided clues for the improvement of visit procedures. A practice visit and feedback by a non-physician observer is more appreciated than a visit and feedback by a colleague. A practice visit with the VIP by a non-physician observer is a simple, easy, and well accepted method for assessing practice management.


The MAPP research network: a novel study of urologic chronic pelvic pain syndromes.
Urologic chronic pelvic pain syndrome (UCPPS) may be defined to include interstitial cystitis/bladder pain syndrome (IC/BPS) and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). The hallmark symptom of UCPPS is chronic pain in the pelvis, urogenital floor, or external genitalia often accompanied by lower urinary tract symptoms. Despite numerous past basic and clinical research studies there is no broadly identifiable organ-specific pathology or understanding of etiology or risk factors for UCPPS, and diagnosis relies primarily on patient reported symptoms. In addition, there are no generally effective therapies. Recent findings have, however, revealed associations between UCPPS and "centralized" chronic pain disorders, suggesting UCPPS may represent a local manifestation of more widespread pathology in some patients. Here, we describe a new and novel effort initiated by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) of the U.S. National Institutes of Health (NIH) to address the many long standing questions regarding UCPPS, the Multidisciplinary Approach to the Study of Chronic Pelvic Pain (MAPP) Research Network. The MAPP Network approaches UCPPS in a systemic manner, in which the interplay between the genitourinary system and other physiological systems is emphasized. The network's study design expands beyond previous research, which has primarily focused on urologic organs and tissues, to utilize integrated approaches to define patient phenotypes, identify clinically-relevant subgroups, and better understand treated natural history and pathophysiology. Thus, the MAPP Network provides an unprecedented, multi-layered characterization of UCPPS. Knowledge gained is expected to provide important insights into underlying pathophysiology, a foundation for better segmenting patients for future clinical trials, and ultimately translation into improved clinical management. In addition, the MAPP Network's integrated multi-disciplinary research approach may serve as a model for studies of urologic and non-urologic disorders that have proven refractory to past basic and clinical study. ClinicalTrials.gov identifier: NCT01098279 "Chronic Pelvic Pain Study of Individuals with Diagnoses or Symptoms of Interstitial Cystitis and/or Chronic Prostatitis (MAPP-EP)".


Horseshoe lung associated with scimitar syndrome and pleural lipoma.
Horseshoe lung is a rare congenital anomaly characterized by the presence of unilateral pulmonary hypoplasia and a midline isthmus bridging the right and the left lung. Almost all cases of horseshoe lung are associated with the scimitar syndrome. The scimitar syndrome, sometimes called the congenital pulmonary venolobar syndrome, is a rare but well-described constellation of cardio-pulmonary anomalies. The characteristic abnormality is anomalous pulmonary venous return from a part of or the entire right lung to the inferior vena cava. We present a case of horseshoe lung associated with scimitar syndrome and pleural lipoma in an adult admitted for acute chest pain and chronic dyspnea.


Anhydrous Ammonia Chemical Release - Lake County, Illinois, April 2019.
On April 25, 2019, a farm tractor towing two 2-ton ammonia tanks on a county road in Lake County, Illinois, experienced a mechanical failure that resulted in the release of anhydrous ammonia, a colorless, pungent, irritating gas that can cause severe respiratory and ocular damage (1). Approximately 80% of anhydrous ammonia produced in the United States is used as a fertilizer in agriculture (1). Eighty-three persons, including first responders, motorists, and neighborhood residents, were evaluated at area hospitals because of exposure to the gas. Two weeks after the release, the Agency for Toxic Substances and Disease Registry (ATSDR) and CDC's National Center for Environmental Health (NCEH) collaborated with the Lake County Health Department and the Illinois Department of Public Health on an investigation using ATSDR's Assessment of Chemical Exposures program to describe the release, review the emergency response, and determine health effects associated with the exposure. First responders, community residents, and hospital personnel reported communication challenges related to the nature of the gas release and effective protective measures. Among the 83 persons evaluated at six area hospitals for effects of the chemical release, 14 (17%) were hospitalized, including eight (10%) who were admitted to the intensive care unit (ICU), seven (8%) of whom required endotracheal intubation and mechanical ventilation; no deaths occurred. In addition, ICU health care providers experienced symptoms of secondary exposure. The National Institute for Occupational Safety and Health's Emergency Responder Health Monitoring and Surveillance Program has specific recommendations and tools to protect responders during all phases of a response (2). Hospitals also need to review institutional policies and procedures for chemical mass casualty events, including decontamination (3). Prompt and correct identification of hazardous material (hazmat) events, and clear communication among responding entities, including on-scene and hospital responders, is important to ensure effective response after a chemical release.


An effortless procedure to align the local frame of an inertial measurement unit to the local frame of another motion capture system.
Inertial measurement units (IMUs) offer great opportunities to analyze segmental and joints kinematics. When combined with another motion capture system (MCS), for example, to validate new IMU-based applications or to develop mixed systems, it is necessary to align the local frame of the IMU sensors to the local frame of the MCS. Currently, all alignment methods use landmarks on the IMU's casing. Therefore, they can only be used with well-documented IMUs and they are prone to error when the IMU's casing is small. This study proposes an effortless procedure to align the local frame of any IMU to the local frame of any other MCS able to measure the orientation of its local frame. The general concept of this method is to derive the gyroscopic angles for both devices during an alignment movement, and then to use an optimization algorithm to calculate the alignment matrix between both local frames. The alignment movement consists of rotations around three more or less orthogonal axes and it can easily be performed by hands. To test the alignment procedure, an IMU and a magnetic marker were attached to a plate, and 20 alignment movements were recorded. The maximum errors of alignment (accuracy±precision) were 1.02°±0.32° and simulations showed that the method was robust against noise that typically affect IMUs. In conclusion, this study describes an efficient alignment procedure that is quick and easy to perform, and that does not require any alignment device or any knowledge about the IMU casing.


Neuroinflammation in glaucoma: A new opportunity.
Mounting evidence suggests neuroinflammation is a key process in glaucoma, yet the precise roles are not known. Understanding these complex processes, which may also be a key in other common neurodegenerations such as Alzheimer's disease, will lead to targeted therapeutics for a disease that affects as many as 80 million people worldwide. Here, we define neuroinflammation as any immune-relevant response by a variety of cell types including astrocytes, microglia, and peripherally derived cells occurring in the optic nerve head and/or retina. In this review article, we first discuss clinical evidence for neuroinflammation in glaucoma and define neuroinflammation in glaucoma. We then review the inflammatory pathways that have been associated with glaucoma. Finally, we set out key research directions that we believe will greatly advance our understanding of the role of neuroinflammation in glaucoma. This review arose from a discussion of neuroinflammation in glaucoma at the 2015 meeting of The Lasker/IRRF Initiative for Innovation in Vision Science. This manuscript sets out to summarize one of these sessions; "Inflammation and Glaucomatous Neurodegeneration", as well as to review the current state of the literature surrounding neuroinflammation in glaucoma.


Pyoderma gangrenosum in an ulcerative colitis patient during treatment with vedolizumab responded favorably to adsorptive granulocyte and monocyte apheresis.
Pyoderma gangrenosum (PG) is an extra-intestinal skin lesion in inflammatory bowel disease (IBD) as is erythema nodosum. Vedolizumab (VED) is a monoclonal antibody that targets α4β7 integrin and has an intestinal selective mechanism. Despite good therapeutic effects on colitis, the effect on extra-intestinal manifestations (EIMs) remains unclear. Here we report a case of ulcerative colitis complicated by PG during treatment with VED, which was successfully treated with prednisolone in combination with adsorptive granulocyte and monocyte apheresis (GMA). The patient was a 50-year-old woman with a past medical history of extensive ulcerative colitis managed by golimumab (GLM). She developed flare symptoms due to loss of response to GLM, and treatment was switched to VED. Her gastrointestinal symptoms were improved with VED treatment with less frequent bowel movements. However, infiltrative erythema with pain appeared on the right lower leg and right knee, and expanded and gradually ulcerated. Her skin lesions were treated with corticosteroid, but showed poor improvement. Therefore, granulocyte and monocyte apheresis (GMA) treatment was administered in combination with prednisolone. After 3 months, the ulcer gradually improved, and at the time of this writing, the eruptions were nearly replaced by epithelial tissue. This case study showed that patients with UC and EIMS may respond well to combination therapy of VED and GMA. GMA has a very favorable safety profile. On the other hand, the causal connection between VED and PG is still unclear. We believe that a combination therapy involving VED and GMA in IBD patients with EIMs warrants consideration.


Comparing muscle temperature during static and dynamic squatting with and without whole-body vibration.
The aim of this study was to investigate the influence of shallow dynamic squatting (DS) versus static squatting (SS) with or without concurrent side-to-side alternating whole-body vibration (WBV) on vastus lateralis temperature and cardiovascular stress as indicated by heart rate (HR). Ten participants (five men, five women) participated in four interventions [DS with WBV (DS+), DS without WBV (DS-), SS with WBV (SS+), SS without WBV (SS-)] 48 h apart, in a randomized order. The interventions were preceded by a approximately 20-min rest period, consisted of 10 mins with or without WBV (26 or 0 Hz) with SS (40 degrees of knee flexion) or DS (55 degrees of knee flexion, at a cadence of 50 bpm) where SS+ and DS- were metabolically matched. Muscle (T(m)), core (T(c)), skin temperature (T(sk)), HR and VO(2) were recorded during each intervention. For T(m), there was a time (P<0.01) and WBV (P<0.01) effect but no squat effect was evident, and there was time xWBV interaction effect (P<0.01). In all four interventions, the work load was too low to cause cardiovascular stress. Instead normal, moderate physiological effects of exercise on autonomic control were observed as indicated by HR; there were no significant increases in T(sk) or T(c). There appears to be no benefit in performing an unloaded, shallow DS+ at a tempo of 50 bpm as T(m,) HR, VO(2) are likely to be increased by the same amount and rate without WBV. However, combining SS with WBV could be advantageous to rapidly increasing soft tissue temperature prior to performing rehabilitation exercises when dynamic exercise cannot be performed.


Protective effects of transforming growth factor β2 in intestinal epithelial cells by regulation of proteins associated with stress and endotoxin responses.
Transforming growth factor (TGF)-β2 is an important anti-inflammatory protein in milk and colostrum. TGF-β2 supplementation appears to reduce gut inflammatory diseases in early life, such as necrotizing enterocolitis (NEC) in young mice. However, the molecular mechanisms by which TGF-β2 protects immature intestinal epithelial cells (IECs) remain to be more clearly elucidated before interventions in infants can be considered. Porcine IECs PsIc1 were treated with TGF-β2 and/or lipopolysaccharide (LPS), and changes in the cellular proteome were subsequently analyzed using two-dimensional gel electrophoresis-MS and LC-MS-based proteomics. TGF-β2 alone induced the differential expression of 13 proteins and the majority of the identified proteins were associated with stress responses, TGF-β and Toll-like receptor 4 signaling cascades. In particular, a series of heat shock proteins had similar differential trends as previously shown in the intestine of NEC-resistant preterm pigs and young mice. Furthermore, LC-MS-based proteomics and Western blot analyses revealed 20 differentially expressed proteins following treatment with TGF-β2 in LPS-challenged IECs. Thirteen of these proteins were associated with stress response pathways, among which five proteins were altered by LPS and restored by TGF-β2, whereas six were differentially expressed only by TGF-β2 in LPS-challenged IECs. Based on previously reported biological functions, these patterns indicate the anti-stress and anti-inflammatory effects of TGF-β2 in IECs. We conclude that TGF-β2 of dietary or endogenous origin may regulate the IEC responses against LPS stimuli, thereby supporting cellular homeostasis and innate immunity in response to bacterial colonization, and the first enteral feeding in early life.


Voluntary eye movements direct attention on the mental number space.
Growing evidence suggests that orienting visual attention in space can influence the processing of numerical magnitude, with leftward orienting speeding up the processing of small numbers relative to larger ones and the converse for rightward orienting. The manipulation of eye movements is a convenient way to direct visuospatial attention, but several aspects of the complex relationship between eye movements, attention orienting and number processing remain unexplored. In a previous study, we observed that inducing involuntary, reflexive eye movements by means of optokinetic stimulation affected number processing only when numerical magnitude was task relevant (i.e., during magnitude comparison, but not during parity judgment; Ranzini et al., in J Cogn Psychol 27, 459-470, (2015). Here, we investigated whether processing of task-irrelevant numerical magnitude can be modulated by voluntary eye movements, and whether the type of eye movements (smooth pursuit vs. saccades) would influence this interaction. Participants tracked with their gaze a dot while listening to a digit. The numerical task was to indicate whether the digit was odd or even through non-spatial, verbal responses. The dot could move leftward or rightward either continuously, allowing tracking by smooth pursuit eye movements, or in discrete steps across a series of adjacent locations, triggering a sequence of saccades. Both smooth pursuit and saccadic eye movements similarly affected number processing and modulated response times for large numbers as a function of direction of motion. These findings suggest that voluntary eye movements redirect attention in mental number space and highlight that eye movements should play a key factor in the investigation of number-space interactions.


Microalgae population dynamics growth with AnMBR effluent: effect of light and phosphorus concentration.
The aim of this study was to evaluate the effect of light intensity and phosphorus concentration on biomass growth and nutrient removal in a microalgae culture and their effect on their competition. The photobioreactor was continuously fed with the effluent from an anaerobic membrane bioreactor pilot plant treating real wastewater. Four experimental periods were carried out at different light intensities (36 and 52 μmol s-1 m-2) and phosphorus concentrations (around 6 and 15 mgP L-1). Four green algae - Scenedesmus, Chlorella, Monoraphidium and Chlamydomonas- and cyanobacterium were detected and quantified along whole experimental period. Chlorella was the dominant species when light intensity was at the lower level tested, and was competitively displaced by a mixed culture of Scenedesmus and Monoraphidium when light was increased. When phosphorus concentration in the photobioreactor was raised up to 15 mgP L-1, a growth of cyanobacterium became the dominant species in the culture. The highest nutrient removal efficiency (around 58.4 ± 15.8% and 96.1 ± 16.5% of nitrogen and phosphorus, respectively) was achieved at 52 μmol s-1 m-2 of light intensity and 6.02 mgP L-1 of phosphorus concentration, reaching about 674 ± 86 mg L-1 of volatile suspended solids. The results obtained reveal how the light intensity supplied and the phosphorus concentration available are relevant operational factors that determine the microalgae species that is able to predominate in a culture. Moreover, changes in microalgae predominance can be induced by changes in the growth medium produced by the own predominant species.


Aboveground overyielding in a mixed temperate forest is not explained by belowground processes.
The relationship between forest productivity and tree species diversity has been described in detail, but the underlying processes have yet to be identified. One important issue is to understand which processes are at the origin of observed aboveground overyielding in some mixed forests. We used a beech-maple plantation exhibiting aboveground overyielding to test whether belowground processes could explain this pattern. Soil cores were collected to determine fine root (FR) biomass and vertical distribution. Correlograms were used to detect spatial arrangement. Near-infrared reflectance spectroscopy was used to identify the tree species proportion in the FR samples and spatial root segregation. An isotopic approach was used to identify water acquisition patterns. The structure and the composition of the ectomycorrhizal fungal community were determined by high-throughput sequencing of DNA in the soil samples. We found no spatial pattern for FR biomass or for its vertical distribution along the gradients. No vertical root segregation was found, as FR density for both species decreased with depth in a similar way. The two species displayed similar vertical water acquisition profiles as well, mainly absorbing water from shallow soil layers; hence, niche differentiation for water acquisition was not highlighted here. Significant alterations in the fungal community compositions were detected in function of the percentage of maple in the vicinity of beech. Our findings do not support the commonly suggested drivers of aboveground overyielding in species-diverse forests and suggest that competition reduction or between-species facilitation of belowground resource acquisition may not explain the observed aboveground overyielding.


Whole-body vibration training increases vertical jump height in a dance population.
Whole-body vibration (WBV) training has gained popularity with the strength and conditioning environment. Previous research reported improvements in strength and power after WBV interventions in untrained individuals with little hypertrophical development of the muscles, suggesting that the improvements are because of neuromuscular adaptations. The present intervention study recruited moderately trained individuals that have jumping as integral to their training. Participants were randomly allocated to an intervention or control group. The intervention group was exposed to WBV at 35 Hz for 5 minutes twice a week, whereas the control group was exposed to a similar isometric contraction stress. Results indicated that after a 6-week intervention, vertical jump height increased significantly (p < 0.05) in the intervention group than in the control group. The study concluded that WBV was able to maintain vertical jump height with very little intervention time (10 minutes a week) and therefore should be considered as an intervention form for maintaining jump performance.