Hotspots of root-exuded amino acids are created within a rhizosphere-on-a-chip.
The rhizosphere is a challenging ecosystem to study from a systems biology perspective due to its diverse chemical, physical, and biological characteristics. In the past decade, microfluidic platforms (e.g. plant-on-a-chip) have created an alternative way to study whole rhizosphere organisms, like plants and microorganisms, under reduced-complexity conditions. However, in reducing the complexity of the environment, it is possible to inadvertently alter organism phenotype, which biases laboratory data compared to in situ experiments. To build back some of the complexity of the rhizosphere in a fully-defined, parameterized approach we have developed a rhizosphere-on-a-chip platform that mimics the physical structure of soil. We demonstrate, through computational simulation, how this synthetic soil structure can influence the emergence of molecular "hotspots" and "hotmoments" that arise naturally from the plant's exudation of labile carbon compounds. We establish the amenability of the rhizosphere-on-a-chip for long-term culture of Brachypodium distachyon, and experimentally validate the presence of exudate hotspots within the rhizosphere-on-a-chip pore spaces using liquid microjunction surface sampling probe mass spectrometry.


Commensal Neisseria cinerea outer membrane vesicles as a platform for the delivery of meningococcal and gonococcal antigens to the immune system.
An affordable, accessible, and broadly protective vaccine is required to tackle the re-occurring bacterial meningococcal epidemics in Sub-Saharan Africa as well as an effective control of multi-drug resistant strains of gonococcus. Outer membrane vesicles (OMVs) secreted from Gram-negative bacteria represent an attractive platform for antigen delivery to the immune system and therefore for development of multi-component vaccines. In this study, we describe the generation of modified OMVs (mOMVs) from commensal biosafety-level 1 (BSL-1) Neisseria cinerea ATCC® 14685TM, which is phylogenetically close to the pathogenic bacteria Neisseria meningitidis and Neisseria gonorrhoeae. mOMVs were prepared from N. cinerea engineered to express heterologous antigens from N. meningitidis (factor H binding protein (fHbp) and Neisseria Heparin Binding Antigen (NHBA-2)) and from N. gonorrhoeae (NHBA-542). Mice immunised with the mOMVs produced antibodies against fHbp and NHBA. The work indicates that mOMV from N. cinerea can be used as a platform to induce immune responses against antigens involved in the protective immune response against meningococcal and gonococcal diseases.


Application of the red fluorescent protein mCherry in mycelial labeling and organelle tracing in the dermatophyte Trichophyton mentagrophytes.
Trichophyton mentagrophytes is a fungus that causes skin disease in humans and other animals worldwide. Studies on molecular biology and fluorescent labeling of the fungus are limited. Here, we applied mCherry for the first time in T. mentagrophytes to label the fungus and its organelles. We constructed four expression vectors of mCherry or mCherry fusions containing a variety of resistance markers and promoters, which were then integrated, together with two previous mCherry expression vectors, in T. mentagrophytes via Agrobacterium tumefaciens-mediated transformation (AtMT). The resulting transformants emitted bright red fluorescence. We used the histone protein H2B and the peroxisome targeting signal 1 (PTS1) peptide to target the nucleus and peroxisomes, respectively, in T. mentagrophytes. In the transformants expressing mCherry-fused H2B, the fluorescence was distinctly localized to the nuclei in hyphae, spores and the fungal cells in infected animal tissue. In the T. mentagrophytes transformants where the peroxisome was targeted, the mCherry was present as small dots (0.2-1 μm diameter) throughout the spores and the hyphae. We also constructed a T. mentagrophytes AtMT library containing more than 1000 hygromycin-resistant transformants that were genetically stable. Our results provide useful tools for further investigations on molecular pathogenesis of T. mentagrophytes.


Effects of two bisdioxopiperazines on mouse B-and T-cell function.
The immunosuppresive effects of the bisdioxopiperazines ICRF 154 and ICRF 159 on the function of bone marrow-derived lymphocytes (B cells) and thymusderived lymphocytes (T cells) were assessed and compared with those of cyclophosphamide (CPA). Mice given foreign erythrocytes with any of these drugs for 3 or 5 days showed suppressed antibody responses due to the inhibition of thymus-derived cooperating lymphocyte (T-helper cell) priming and inactivation of B cells in the spleen. In contrast, 3 days of drug treatment after the injection of allogeneic tumor cells only partially inhibited T cell-dependent cytotoxicity. Moreover, when irradiated tumor cells were given for 3 days with CPA or ICRF 154, enhancement rather than inhibition of cytotoxicity was the usual response. However, with both types of immunization, no thymus-derived cytotoxic lymphocytes (T killer cells) could be generated after prolonged treatment (6 daily injections) with any of the 3 drugs. Administration of drugs before antigen also resulted in selective drug action, i.e., a relative increase in the proportion of thy-1-positive cells in the spleen. After challenge in vivo or in vitro with erythrocyte antigens, the plaque-forming cell responses of these spleens were raised up to twofold, probably because they had higher T-helper cell activity than untreated controls. Similar pretreatment before immunization with allogeneic tumor cells also led to enhanced T-killer cell activity, but only in ICRF 154- and CPA-treated mice. These observations suggest that under certain conditions, the bisdioxopiperazines and CPA have selective effects on B-cell rather than T-cell function.


An efficient and anonymous buyer-seller watermarking protocol.
For the purpose of deterring unauthorized duplication and distribution of multimedia contents, a seller may insert a unique digital watermark into each copy of the multimedia contents to be sold. When an illegal replica is found in the market sometime later, the seller can determine the responsible distributor by examining the watermark embedded. However, the accusation against the charged distributor, who was the buyer in some earlier transaction, is objectionable because the seller also has access to the watermarked copies and, hence, is able to release such a replica on her own. In this paper, a watermarking protocol is proposed to avoid such difficulties, known as the customer's right problem, in the phase of arbitration. The proposed watermarking protocol also provides a fix to Memon and Wong's scheme by solving the unbinding problem. In addition, the buyer is no longer required to contact the watermark certification authority during transactions, and the anonymity of the buyer can be retained through a trusted third party. The result is an efficient and anonymous buyer-seller watermarking protocol.


Mapping the polarization pattern of plasmon modes reveals nanoparticle symmetry.
We study the wavelength and polarization dependent plasmon resonances of single silver and gold nanorods, triangles, cubes, and dimers with a novel single particle spectroscopy method (RotPOL). In RotPOL, a rotating wedge-shaped polarizer encodes the full polarization information of each particle within one image. This reveals the symmetry of the particles and their plasmon modes, allows analyzing inhomogeneous samples and the monitoring of particle shape changes during growth in situ.


Responses to soil pH gradients of inorganic phosphate solubilizing bacteria community.
Soil pH is commonly considered a dominant factor affecting the function of microbiota. Few studies, however, have focused on communities of bacteria able to solubilize inorganic phosphate (iPSB), which are important for the mobilization of soil phosphorus (P), because finding an effective method to assess the abundance and diversity of iPSB communities is difficult. We used a newly reported method of database alignment and quantified the gene pqqC to analyze the compositions of iPSB communities from five soils with pH gradients ranging from 4 to 8. The iPSB community structure differed significantly between these soil types. Among iPSB community, Bacillus was the dominant genus, followed by Arthrobacter and Streptomyces. A redundancy analysis indicated that soil pH was the most important of 15 soil factors and their pairwise interactions, accounting for 5.12% of the variance. The abundance of the iPSB communities increased with pH within the gradients which was confirmed by experimental adjustment of pH, suggesting that the defect P status in high pH soil was speculated as the driving force of iPSB community population. Our study demonstrated the dominant role of soil pH on the iPSB community, which may contribute to the understanding the possible mechanism of microbial P mobilization for better improvement of P use-efficiency.


Chronic D-amphetamine induces sexually dimorphic effects on locomotion, recognition memory, and brain monoamines.
While acute and chronic D-amphetamine (AMPH) treatments produce greater scores for locomotor activity in female rats in comparison with male rats, little is known about AMPH-induced gender differences on cognition. The objectives of the present study were to (1) investigate during a withdrawal period following chronic AMPH treatment whether performance of two memory tasks, object recognition (OR) and object placement (OP) are altered, and (2) determine if an AMPH challenge dose after a withdrawal period amplifies previously reported gender differences in locomotor activity and neurochemistry. Sprague-Dawley male and female adult rats were included in a chronic AMPH treatment (10 injections, 1 every other day; males: 3 mg/kg, females 2.6 mg/kg). Locomotor activity was quantified (acute, chronic, and after a 16-day withdrawal period). Neurotransmitter levels in brain areas were evaluated after an AMPH challenge dose on the 16th withdrawal day. During the withdrawal period, OR (2- and 4-h delays) was impaired in AMPH-treated males but they did not show any impairment in OP; AMPH females also showed impairments in OR (only 4-h delay). AMPH females showed more locomotion after acute and chronic treatment but AMPH-induced hyperactivity was comparable for females and males after a challenge dose. Following a challenge dose of AMPH after a withdrawal period, gender differences in dopaminergic and serotonergic neurotransmission in the striatum were found. These gender differences elicited by AMPH in monoaminergic pathways may be related to sex differences on behavioral components involved in locomotion and OR memory.


Strain and stress variations in the human amniotic membrane and fresh corpse autologous sciatic nerve anastomosis in a model of sciatic nerve injury.
A 10-mm long sciatic nerve injury model was established in fresh normal Chinese patient cadavers. Amniotic membrane was harvested from healthy maternal placentas and was prepared into multilayered, coiled, tubular specimens. Sciatic nerve injury models were respectively anastomosed using the autologous cadaveric sciatic nerve and human amniotic membrane. Tensile test results showed that maximal loading, maximal displacement, maximal stress, and maximal strain of sciatic nerve injury models anastomosed with human amniotic membrane were greater than those in the autologous nerve anastomosis group. The strain-stress curves of the human amniotic membrane and sciatic nerves indicated exponential change at the first phase, which became elastic deformation curves at the second and third phases, and displayed plastic deformation curves at the fourth phase, at which point the specimens lost their bearing capacity. Experimental findings suggested that human amniotic membranes and autologous sciatic nerves exhibit similar stress-strain curves, good elastic properties, and certain strain and stress capabilities in anastomosis of the injured sciatic nerve.


Structural insight into negative DNA supercoiling by DNA gyrase, a bacterial type 2A DNA topoisomerase.
Type 2A DNA topoisomerases (Topo2A) remodel DNA topology during replication, transcription and chromosome segregation. These multisubunit enzymes catalyze the transport of a double-stranded DNA through a transient break formed in another duplex. The bacterial DNA gyrase, a target for broad-spectrum antibiotics, is the sole Topo2A enzyme able to introduce negative supercoils. We reveal here for the first time the architecture of the full-length Thermus thermophilus DNA gyrase alone and in a cleavage complex with a 155 bp DNA duplex in the presence of the antibiotic ciprofloxacin, using cryo-electron microscopy. The structural organization of the subunits of the full-length DNA gyrase points to a central role of the ATPase domain acting like a 'crossover trap' that may help to sequester the DNA positive crossover before strand passage. Our structural data unveil how DNA is asymmetrically wrapped around the gyrase-specific C-terminal β-pinwheel domains and guided to introduce negative supercoils through cooperativity between the ATPase and β-pinwheel domains. The overall conformation of the drug-induced DNA binding-cleavage complex also suggests that ciprofloxacin traps a DNA pre-transport conformation.


A CRISPR-Cas9-based reporter system for single-cell detection of extracellular vesicle-mediated functional transfer of RNA.
Extracellular vesicles (EVs) form an endogenous transport system for intercellular transfer of biological cargo, including RNA, that plays a pivotal role in physiological and pathological processes. Unfortunately, whereas biological effects of EV-mediated RNA transfer are abundantly studied, regulatory pathways and mechanisms remain poorly defined due to a lack of suitable readout systems. Here, we describe a highly-sensitive CRISPR-Cas9-based reporter system that allows direct functional study of EV-mediated transfer of small non-coding RNA molecules at single-cell resolution. Using this CRISPR operated stoplight system for functional intercellular RNA exchange (CROSS-FIRE) we uncover various genes involved in EV subtype biogenesis that play a regulatory role in RNA transfer. Moreover we identify multiple genes involved in endocytosis and intracellular membrane trafficking that strongly regulate EV-mediated functional RNA delivery. Altogether, this approach allows the elucidation of regulatory mechanisms in EV-mediated RNA transfer at the level of EV biogenesis, endocytosis, intracellular trafficking, and RNA delivery.


Acute Metabolic Influences on the Natriuretic Peptide System in Humans.
The cardiac natriuretic peptides (NPs), atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP), have central roles in sodium and blood pressure regulation. Extracardiac factors (e.g., obesity and diabetes) influence NP production, potentially altering cardiovascular responses to volume and pressure stress. This study examined the effects of acute carbohydrate intake on the NP system in humans, and investigated underlying mechanisms. Normotensive subjects (N = 33) were given a high-carbohydrate shake. Venous blood was sampled to measure N-terminal (NT)-proANP and NT-proBNP levels. Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) and HepG2 cells were treated with glucose, and expression levels of NPs and micro ribonucleic acid 425 (miR-425), a negative regulator of ANP, were examined. The role of nuclear factor kappa B (NF-κB) in the glucose-mediated effects was investigated using a NF-κB inhibitor and expression plasmids encoding NF-κB subunits. We observed a 27% reduction in the levels of circulating NT-proANP (p < 0.001, maximal at 6 h) after carbohydrate challenge, with no effect on NT-proBNP levels in our human subjects. Glucose treatment of hESC-CMs for 6 h and 24 h increased levels of the primary transcript of miR-425 (pri-miR-425) and mature miR-425. A corresponding decrease in NPPA messenger RNA levels was also observed at both time points. Overexpression of NF-κB subunits in H9c2 cardiomyocytes increased miR-425 levels, whereas inhibition of NF-κB abrogated the glucose-mediated increase in pri-miR-425 levels in HepG2 cells. Acute carbohydrate challenge is associated with a reduction in ANP production. The mechanism appears to involve a glucose-induced increase in the expression of miR-425, mediated by NF-κB signaling.


Advancing the application of systems thinking in health: South African examples of a leadership of sensemaking for primary health care.
New forms of leadership are required to bring about the fundamental health system changes demanded by primary health care (PHC). Using theory about complex adaptive systems and policy implementation, this paper considers how actors' sensemaking and the exercise of discretionary power currently combine to challenge PHC re-orientation in the South African health system; and provides examples of leadership practices that promote sensemaking and power use in support of PHC. The paper draws on observational, interview, and reflective data collected as part of the District Innovation and Action Learning for Health Systems Development (DIALHS) project being implemented in Cape Town, South Africa. Undertaken collaboratively between health managers and researchers, the project is implemented through cycles of action-learning, including systematic reflection and synthesis. It includes a particular focus on how local health managers can better support front line facility managers in strengthening PHC. The results illuminate how the collective understandings of staff working at the primary level - of their working environment and changes within it - act as a barrier to centrally-led initiatives to strengthen PHC. Staff often fail to take ownership of such initiatives and experience them as disempowering. Local area managers, located between the centre and the service frontline, have a vital role to play in providing a leadership of sensemaking to mediate these challenges. Founded on personal values, such leadership entails, for example, efforts to nurture PHC-aligned values and mind-sets among staff; build relationships and support the development of shared meanings about change; instil a culture of collective inquiry and mutual accountability; and role-model management practices, including using language to signal meaning. PHC will only become a lived reality within the South African health system when frontline staff are able to make sense of policy intentions and incorporate them into their everyday routines and practices. This requires a leadership of sensemaking that enables front line staff to exercise their collective discretionary power in strengthening PHC. We hope this theoretically-framed analysis of one set of experiences stimulates wider thinking about the leadership needed to sustain primary health care in other settings.


Sex differences in mental rotation strategy.
When humans decide whether two visual stimuli are identical or mirror images of each other and one of the stimuli is rotated with respect to the other, the time discrimination takes usually increases as a rectilinear function of the orientation disparity. On the average, males perform this mental rotation at a faster angular speed than females. This experiment required the rotation of both mirror-image-different and non-mirror-different stimuli. The polygonal stimuli were presented in either spatially unfiltered, high-pass or low-pass filtered versions. All stimulus conditions produced mental rotation-type effects but with graded curvilinear trends. Women rotated faster than men under all conditions, an infrequent outcome in mental rotation studies. Overall, women yielded more convexly curvilinear response functions than men. For both sexes the curvilinearity was more pronounced under the non-mirror-different, low-pass stimulus condition than under the mirror different, high-pass stimulus condition. The results are considered as supporting the occurrence of two different mental rotation strategies and as suggesting that the women were predisposed to use efficiently an analytic feature rotation strategy, while the men were predisposed to employ efficiently a holistic pattern rotation strategy. It is argued that the overall design of this experiment promoted the application of an analytic strategy and thus conferred an advantage to the female participants.


Necroptosis in Acute Kidney Injury.
Regulated necrosis is an expanding research field with important implications for acute kidney injury (AKI). A focused review of the evolving evidence for necroptosis in AKI, one of several forms of regulated necrosis defines the known and unknown. A literature search was performed in PUBMED and ScienceDirect between January 1957 and April 2018 using the following keywords: "acute kidney injury," "necrosis," "necroptosis," "necroinflammation." The necroptosis signaling cascade involves a number of proteins including receptor-interacting protein-1 (RIPK1), RIPK3, and mixed lineage kinase domain-like pseudokinase (MLKL) as well as the MLKL regulator RGMb. The existing experimental evidence in AKI based on mice with genetic deletions of these proteins, more or less specific inhibitory compounds, and diverse experimental AKI models is reviewed. There is broad consistency suggesting a role for necroptosis in AKI, but some studies report divergent evidence potentially relating to the specific model used and the time point of analysis. Mlkl-deficient mice are currently the most specific and reliable experimental tool to study necroptosis in vivo (in kidney disease). The clinical potential of necroptosis inhibition in AKI is to be evaluated, but conceptual problems in AKI definitions and in complex clinical scenarios remain a concern.


Chitosan-based systems for the delivery of vaccine antigens.
This review discusses the current status of chitosan and its derivatives as adjuvants and delivery systems in vaccine delivery, and their future possibilities and challenges. After a brief introduction to adjuvants and delivery systems, chitosan will be described in detail in regard to vaccine formulation. Applications of chitosan and its derivatives will be reviewed and their proposed mechanisms in the enhancement of immune responses will be discussed.


Ototoxicity of different concentrations povidone-iodine solution applied to the middle ear cavity of rats.
To investigate the ototoxic effects of different concentrations of povidone-iodine solutions applied to the middle ear cavity of rats using distortion product otoacoustic emissions. 24 healthy 3-3.5-month-old adult female Sprague-Dawley rats were randomly divided into three groups. The group A (n = 8 ears) received 5 % povidone-iodine solution to the right ear, the group B (n = 8 ears) received 7.5 % povidone-iodine solution to the right ear and the group C (n = 8 ears) received 10 % povidone-iodine solution to the right ear. All animals received saline solution to the left ear as a control (n = 24 ears). The animals were tested before, 1 and 10 days after solutions administration to the middle ear. The resulting distortion product otoacoustic emissions were evaluated at 1.5, 2, 3, 4, 5, 6, 7, 8, 10 and 12 kHz. Statistically significant reductions in DP-gram amplitudes were noted at high frequencies (7, 8, 10, 12 kHz) in the group A at day 1 but this effect return at day 10. In group B and group C statistically significant differences were recorded for low and high frequencies (1.5, 2, 7, 8, 10, 12 kHz) according to the control group at day 1 and 10. 7.5 and 10 % povidone-iodine showed a significant ototoxic effect on day 1 and 10. But this toxic effect could not be elicited in 5 % povidone-iodine group on day 10. The present study revealed that commercially available high concentration povidone-iodine solution may cause significant ototoxic effects when applied topically through a perforated ear drum in rats. Based on results of this experiment, high concentration povidone-iodine solutions should not be used for preoperative surgical site cleansing for otologic surgery.


Time Trends in Concussion Symptom Presentation and Assessment Methods in High School Athletes.
Concussion rates have increased significantly over the past decade. This may reflect an increase in the knowledge and diagnosis of the symptoms of a concussion rather than a true increase in the incidence. Assessing trends in the way that concussions are presenting to and being identified by clinicians over the same period may provide additional insight into the apparent rise in concussions. To evaluate patterns of change in concussion symptom presentation, diagnostic/evaluation methods, and symptom resolution time reported for United States high school athletes from the 2007-2008 through 2014-2015 academic years. Descriptive epidemiology study. This study is a retrospective analysis of a web-based longitudinal high school sports injury surveillance database (High School RIO [Reporting Information Online]) collected from 2007-2008 through 2014-2015. For each concussion, athletic trainers entered data regarding symptom presentation, resolution time, and diagnostic/evaluation tools utilized. Academic year was the primary exposure in assessing each aim. Time trends were then assessed using linear regression or the Cochran-Armitage test for trends, depending on the outcome distribution. The proportion of concussed athletes presenting with amnesia, loss of consciousness (LOC), and tinnitus significantly decreased from 2007-2008 through 2014-2015, while the proportion presenting with drowsiness, irritability, light sensitivity, and noise sensitivity increased significantly. The use of diagnostic radiography, magnetic resonance imaging, and computed tomography all significantly decreased during the study period, while the use of computerized neurocognitive tests increased. Concussion symptoms took significantly longer to resolve in more recent years. The decrease in what have traditionally been considered severe symptoms (LOC, amnesia) and the increase in what were traditionally considered minor symptoms (drowsiness, irritability, light sensitivity) suggest that clinicians may have a lower threshold in diagnosing sports-related concussions in more recent years. The significant reduction in the use of all forms of diagnostic head imaging demonstrates an increased recognition of concussions as functional disturbances rather than structural abnormalities. Improved concussion education and the nationwide passage of state-level concussion legislation have likely led to the increased recognition of lingering symptoms in athletes with a diagnosed concussion, thereby leading to a longer symptom resolution time.


Exploration of CP12 conformational changes and of quaternary structural properties using electrospray ionization traveling wave ion mobility mass spectrometry.
CP12 is a small chloroplast protein involved in the Benson-Calvin cycle. Since it was demonstrated that the CP12 protein shared different conformational properties between reduced and oxidized states we took advantage of the segregational properties of the Traveling Wave Ion Mobility (TWIM) guide to study subtle conformational changes related to redox changes. Electrospray ionization mass (ESI-MS) spectra of the CP12 protein were recorded in the positive ion mode using an ESI source fitted on a quadrupole time-of-flight (QToF) hybrid mass spectrometer equipped with a TWIM cell (Synapt HDMS G1, Waters Corp., Manchester) under non-denaturing conditions. Non-covalent experiments were performed using the same instrument without the use of the TWIM device. Whatever the CP12 form studied, our results showed that CP12 protein was represented by two conformers in equilibrium that displayed very slight differences. These observations led us to propose that CP12 protein structure is rather undergoing transient subtle structural changes than having two different conformational populations in solution. In addition, using non-denaturing experiments, NAD and CP12 stoichiometry were determined with respect to the GAPDH tetramer and the redox state of CP12. In this study we showed that the use of the segregational property of the ion mobility (TWIM, Synapt G1 HDMS, Waters, Manchester, UK) allowed differentiation of subtle conformational changes between redox states of the CP12 protein. Standard non-denaturing experiments revealed different binding stoichiometry according to the redox state of the CP12 protein.


A neural-network packet switch controller: scalability, performance, and network optimization.
We examine a novel combination of architecture and algorithm for a packet switch controller that incorporates an experimentally implemented optically interconnected neural network. The network performs scheduling decisions based on incoming packet requests and priorities. We show how and why, by means of simulation, the move from a continuous to a discrete algorithm has improved both network performance and scalability. The system's limitations are examined and conclusions drawn as to its maximum scalability and throughput based on today's technologies.


Comparison of (68)Ga-DOTA-Tyr(3)-octreotide and (18)F-fluoro-L-dihydroxyphenylalanine positron emission tomography in neuroendocrine tumor patients.
(68)Ga-DOTA-Tyr3-octreotide positron emission tomography ((68)Ga-DOTA-TOC PET) and (18)F-fluoro-L-dihydroxyphenylalanine PET ((18)F-DOPA PET) are emerging modalities for imaging of neuroendocrine tumors. This study reports our initial experiences with these two PET modalities on initial diagnosis, staging and restaging in NET patients. Fifteen patients with NET underwent both (68)Ga-DOTA-TOC and (18)F-DOPA PET as well as computed tomography (CT). Image findings were compared on a patient-basis (pathological uptake: yes/no) as well as on a lesion-basis. Contrast-enhanced CT and histological follow-up served as gold standard. Furthermore, imaging results were matched with tumor marker levels and quantitative tracer uptake by the tumor lesions. When comparing (68)Ga-DOTA-TOC and (18)F-DOPA PET, each modality showed a sensitivity of 64% and a specificity of 100% on a patient-based analysis. (68)Ga-DOTA-TOC PET and (18)F-DOPA PET showed equal findings in 7 out of 15 patients and disagreement in 8 patients. (68)Ga-DOTA-TOC revealed more metastases than (18)F-DOPA PET in 6 patients, while (18)F-DOPA PET detected more metastases than (68)Ga-DOTA-TOC in 4 patients. By (68)Ga-DOTA-TOC PET, 208 malignant lesions were detected, while by (18)F-DOPA only 86 lesions were found, and in CT 124, respectively. (68)Ga-DOTA-TOC and (18)F-DOPA PET are useful tools in the detection and staging of NET lesions. Our initial results allow the conclusion that (68)Ga-DOTA-TOC PET may have a stronger clinical impact in NET patients, as it does not only offer diagnostic information, but is decisive for the further treatment management, i. e. PRRT, as well.


Ordering of binary colloidal crystals by random potentials.
Structural defects are ubiquitous in condensed matter, and not always a nuisance. For example, they underlie phenomena such as Anderson localization and hyperuniformity, and they are now being exploited to engineer novel materials. Here, we show experimentally that the density of structural defects in a 2D binary colloidal crystal can be engineered with a random potential. We generate the random potential using an optical speckle pattern, whose induced forces act strongly on one species of particles (strong particles) and weakly on the other (weak particles). Thus, the strong particles are more attracted to the randomly distributed local minima of the optical potential, leaving a trail of defects in the crystalline structure of the colloidal crystal. While, as expected, the crystalline ordering initially decreases with an increasing fraction of strong particles, the crystalline order is surprisingly recovered for sufficiently large fractions. We confirm our experimental results with particle-based simulations, which permit us to elucidate how this non-monotonic behavior results from the competition between the particle-potential and particle-particle interactions.


Clinical characteristics and patterns of health deficits of centenarians receiving home care and long-term care services.
Centenarians (persons aged 100years and older) are one of the fastest growing cohorts in countries across the world. With the increasing prevalence of centenarians and growing amount of clinical information in large administrative health databases, it is now possible to more fully characterize the health of this unique and heterogeneous population. This study described patterns of health deficits in the centenarian population receiving care from community-based home care services and long-term care facilities (LTCFs) in Ontario, Canada. All centenarians who received home care and were assessed using the interRAI-Home Care Assessment instrument between 2007 and 2011 (n=1163) and all centenarians who resided in LTCFs between 2005 and 2011 who were assessed using the interRAI Minimum Data Set (MDS 2.0) (n=2228) were included in this study. Bivariate analyses described the centenarian population while K-means clustering analyses were utilized to identify relatively homogeneous subgroups within this heterogeneous population. The 3391 centenarians were aged 100 to 114 (mean age 101.5years ±1.9 SD) and the majority were women (84.7%). Commonly reported deficits included cognitive impairment, physical impairment, and bladder problems. Centenarians residing in LTCFs were significantly more likely than centenarians receiving home care services to report cognitive or functional impairment, or to exhibit symptoms of depression. The commonalities and uniqueness of four clusters of centenarians are described. Although there is great variability, there is also commonality within the centenarian population. Recognizing patterns within the heterogeneity of centenarians is key to providing high-quality person-centered care and to targeting health promotion and intervention strategies.


Health Social Networks of Black Women With Hypertension.
The prevalence of hypertension is 55% among African American/Black women, who have a higher risk for poor health outcomes compared to women from other racial and ethnic groups, in part because of uncontrolled blood pressure. Previous research results suggest that peers may positively influence self-management of chronic conditions like hypertension. However, few studies have described the personal characteristics of peers in the health social networks of Black women. This substudy aimed to examine health social networks and describe the peers' characteristics, as reported by a convenience sample of Black women with hypertension. In this analysis of data from a larger study, 94 Black women with hypertension attending a church conference participated in a cross-sectional, descriptive study. Their mean age was 59 years, and their mean systolic blood pressure was 143 mm Hg. All participants completed a survey to gather data about (a) the characteristics of individuals they discussed health matters with (their peers or health social network) and (b) their perceptions about hypertension status and knowledge of hypertension among the peers in their health social network. Collectively, participants from the larger study named a total of 658 peers who were part of their health social networks; the mean health social network size was six peers. The peers were mostly women, Black, family members, and, on average, 54 years old. The participants discussed hypertension with 71% of the peers, reported that 36% had hypertension, and felt that 67% were somewhat or very knowledgeable about the condition. A small, positive correlation existed between the participants' health social network size (number of peers named) and their systolic blood pressure levels. The health social network peers were similar to those in the larger study, with most of the same gender, race, and age. The findings of this analysis may be used to help practitioners and scientists guide patients in building health social networks for support in self-managing hypertension and conducting future studies to examine the best strategies for developing and using health social networks to improve health outcomes and reduce health disparities.


Sentinel lymph node biopsy in cutaneous melanoma: the WHO Melanoma Program experience.
We report the experience of the World Health Organization (WHO) Melanoma Program concerning sentinel lymph node (SLN) biopsy for detecting patients with occult regional nodal metastases to submit to selective regional node dissection. From February 1994 to August 1998, in 12 centers of the WHO Melanoma Program, 892 SLN biopsies were performed in 829 patients with clinical stage I melanoma (male: 370; female: 459; median age: 50 years old). The location of the primary melanoma was as follows: trunk 35%; lower limbs, 45%; upper limbs, 18%; and head and neck, 2%. Blue dye injection for SLN identification was performed in all cases; preoperative lymphoscintigraphy was done in 440 patients, and an intra-operative probe for a radio-guided biopsy was used in 141 cases. Overall, the SLN identification rate was 88%. In 68% of the patients, only one SLN was identified, whereas two and three or more SLN were detected in 24% and 8% of the remaining cases, respectively. Overall SLN positivity rate was 18%. Intra-operative frozen section examination was performed in 39% of the cases and was helpful in detecting occult localizations only in 47% of the positive SLNs. Distribution of positive cases by primary thickness was as follows: < 1mm: 2%; 1-1.99 mm: 7%; 2-2.99 mm: 13%; and > or = 3 mm: 31%. Positive nonsentinel lymph nodes were found in 22% of cases with positive SLN submitted for selective dissection. No complications due to the procedure were registered. Of 710 patients who were evaluated, 40 (6%) presented a regional nodal relapse after a negative SLN biopsy and underwent a delayed therapeutic dissection. From the 710 enrolled cases, 638 (88.5%) were alive without evidence of disease at the time of this writing. A multivariate analysis showed SLN status as one of the most significant prognostic factors (P = .000) along with thickness (P = .001) and ulceration (P = .015) of primary tumor. These data confirm the feasibility and safety of the SLN technique for selecting patients to submit to a radical node dissection. The data represent the basis for a future trial by the WHO Melanoma Program in this field to evaluate the most appropriate surgical approach for treating patients with occult regional nodal metastases.


Spaceflight impairs antigen-specific tolerance induction in vivo and increases inflammatory cytokines.
The health risks of a dysregulated immune response during spaceflight are important to understand as plans emerge for humans to embark on long-term space travel to Mars. In this first-of-its-kind study, we used adoptive transfer of T-cell receptor transgenic OT-II CD4 T cells to track an in vivo antigen-specific immune response that was induced during the course of spaceflight. Experimental mice destined for spaceflight and mice that remained on the ground received transferred OT-II cells and cognate peptide stimulation with ovalbumin (OVA) 323-339 plus the inflammatory adjuvant, monophosphoryl lipid A. Control mice in both flight and ground cohorts received monophosphoryl lipid A alone without additional OVA stimulation. Numbers of OT-II cells in flight mice treated with OVA were significantly increased by 2-fold compared with ground mice treated with OVA, suggesting that tolerance induction was impaired by spaceflight. Production of proinflammatory cytokines were significantly increased in flight compared with ground mice, including a 5-fold increase in IFN-γ and a 10-fold increase in IL-17. This study is the first to show that immune tolerance may be impaired in spaceflight, leading to excessive inflammatory responses.


The evaluation of a standardized call/recall system for childhood immunizations in Wandsworth, England.
To improve uptake of childhood immunizations in Wandsworth we developed a standardized call/recall system based on parents being sent three reminders and defaulters being referred to a Health Visitor. Thirty-two out of 44 primary care practices in the area implemented the intervention in September 2011. The aim of this study was to evaluate the implementation, delivery and impact on immunization uptake of the new call/recall system. To assess implementation and delivery, a mixed method approach was used including qualitative (structured interviews) and quantitative (data collected at three months post-implementation) assessment. To assess the impact, we used Student's t test to compare the difference in immunization uptake rates between intervention and non-intervention practices before and after implementation. The call/recall system was viewed positively by both parents and staff. Most children due or overdue immunizations were successfully captured by the 1st invitation reminder. After three invitations, between 87.3 % (MMR1) and 92.2 % (pre-school booster) of children identified as due or overdue immunizations successfully responded. Prior to implementation there was no difference in uptake rates between intervention and non-intervention practices. Post-implementation uptake rates for DTaP/IPV/Hib, MMR1, MMR2 and the pre-school booster were significantly greater in the intervention practices. Similar findings were seen for PCV and Hib/MenC boosters, although the differences were not statistically significant at the 5 % level. Following the successful implementation of a standardized call/recall system in Wandsworth, other regions or primary care practices may wish to consider introducing a similar system to help improve their immunization coverage levels.


Dact2 is expressed in the developing ureteric bud/collecting duct system of the kidney and controls morphogenetic behavior of collecting duct cells.
The overall pattern of the developing kidney is set in large part by the developing ureteric bud/collecting duct system, and dysgenesis of this system accounts for a variety of clinically significant renal diseases. Understanding how the behavior of cells in the developing ureteric bud/collecting duct is controlled is therefore important to understanding the normal and abnormal kidney. Dact proteins have recently been identified as cytoplasmic regulators of intracellular signaling. Dact1 inhibits Wnt signaling, and Dact2 inhibits transforming growth factor (TGF)-β signaling. Here, we report that Dact2 is expressed in developing and adult mouse kidneys, specifically in the ureteric bud/collecting duct epithelium, a structure whose morphogenesis is controlled partially by TGF-β. When small interfering RNA is used to knock down Dact2 expression in collecting duct cells, they show some constitutive phospho-Smad2, undetectable in controls, and elevated phospho-Smad2 in response to TGF-β. They also show defective migration and, in a monolayer wound-healing assay, they fail to assemble a leading edge "cable" of actomyosin and advance instead as a disorganized mass of lamellipodium-bearing cells. This effect is seriously exacerbated by exogenous TGF-β, although control cells tolerate it well. In three-dimensional culture, Dact2 knockdown cells form cysts and branching tubules, but the outlines of the cysts made by knockdown cells are ragged rather than smooth and the branching tubules are decorated with many fine spikes not seen in controls. These data suggest Dact2 plays a role in regulating morphogenesis by renal collecting duct cells, probably by protecting cells from overly strong TGF-β pathway activation.


[Focal therapy in urology: kidney cancer].
Focal therapy has gained attention in the treatment of small renal masses (SRM). However, its use is limited due to scarce data on long-term outcomes. The availability of such data is significantly lower as compared to the relevant data on surgery outcomes. At the same time, minimally invasive surgery has seen the development of laparoscopic nephron-sparing surgery and, recently, robot-assisted surgery. Our purpose is to review the possibilities of treatment for SMR with particular attention on focal therapy. Clinical series and comprehensive reviews support safety and mid/long-term efficacy of renal cryoablation or radiofrequency ablation. Comparative studies and meta-analysis outlined oncological inferiority against partial nephrectomy in local tumor control. For smaller and more peripheral lesions, radiofrequency ablation showed best indications than cryoablation. There are significant demographic and tumor differences between patients treated by one or another approach. The correct indication for each treatment seems to be of key importance to achieve the best oncological and functional outcome. Open partial nephrectomy remains the gold standard treatment for PMR, but laparoscopic approaches have been showing similar results.


Transforming growth factor beta 1 prevents cytokine-mediated inhibitory effects and induction of nitric oxide synthase in the RINm5F insulin-containing beta-cell line.
The aim of this study was to examine if the growth factor, transforming growth factor beta 1 (TGF beta 1), could prevent induction of nitric oxide synthase and cytokine-mediated inhibitory effects in the insulin-containing, clonal beta cell line RINm5F. Treatment of RINm5F cells for 24 h with interleukin-1 beta (IL-1 beta) (100 pM) induced expression of nitric oxide synthase and inhibited glyceraldehyde-stimulated insulin secretion. Combinations of IL-1 beta (100 pM), tumour necrosis factor-alpha (100 pM) and interferon-gamma (100 pM) reduced RINm5F cell viability (determined by the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium (MTT) reduction assay) and de novo protein synthesis, as measured by incorporation of radiolabelled amino acids into perchloric acid-precipitable protein. Pretreatment of RINm5F cells with TGF beta 1 (10 pM) for 18 or 24 h, prior to the addition of either IL-1 beta or combined cytokines, prevented cytokine-induced inhibition of insulin secretion, protein synthesis and the loss of cell viability. TGF beta 1 pretreatment inhibited cytokine-induced expression and activity of nitric oxide synthase in RINm5F cells as determined by Western blotting and by cytosolic conversion of radiolabelled arginine into labelled citrulline and nitric oxide. Chemically generated superoxide also induced expression of nitric oxide synthase possibly due to direct activation of the nuclear transcription factor NF kappa B, an effect prevented by both an antioxidant and TGF beta 1 pretreatment. In conclusion, the mechanism of action of TGF beta 1 in blocking cytokine inhibitory effects was by preventing induction of nitric oxide synthase.


The relations of ego-resiliency and emotion socialization to the development of empathy and prosocial behavior across early childhood.
The present study explored early personality and environmental predictors of the development of young children's empathy, as well as relations of empathy to prosocial behavior with peers at a later age. How children manage their own emotions and behaviors when under stress--their ego-resiliency--would be expected to affect their responses to others' emotions. Also, socialization experiences, such as the quality of parenting behaviors, have been associated with individual differences in empathy-related responding. We examined whether mothers' emotion socialization practices and children's ego-resiliency at 18 months predicted initial levels and change in empathy across five time points (24, 30, 42, 48, and 54 months; N = 242), and whether empathy in turn predicted prosocial behavior with peers at 72/84 months of age. Ego-resiliency and mothers' expressive encouragement both uniquely predicted the intercept of empathy. Boys' empathy was lower than girls' but improved more with age. Initial levels and growth of empathy positively predicted later prosocial behavior. Children's ego-resiliency predicted the slope of empathy at near significance (p = .054). We also found that the intercept of empathy mediated the relation between ego-resiliency and prosocial behavior as well as the relation between mothers' expressive encouragement and prosocial behavior. These findings suggest that both parenting and personality characteristics are relevant to the development of empathy during early childhood and might contribute to children's later prosocial behavior with peers.


Systemic alterations in plasma proteins from women with chronic widespread pain compared to healthy controls: a proteomic study.
Chronic widespread pain (CWP) is a complex pain condition that is difficult to treat. The prevalence of CWP approximates ~10% of the general population, with higher prevalence in women. Lack of understanding of molecular mechanisms has been a challenge for diagnosis and treatment of chronic pain. The aim of this study was to explore the systemic protein changes in CWP compared to those in healthy controls (CON). By applying 2-dimensional gel electrophoresis, we analyzed the protein pattern of plasma samples from women with CWP (n=16) and healthy women (n=23). The proteomic data were analyzed using multivariate statistical models, and altered proteins were identified using mass spectrometry. The proteome analysis was further validated by gel-free Western blot. Multivariate statistical data analysis of quantified proteins revealed 22 altered proteins in women with CWP, compared to CON group. Many of the identified proteins are previously known to be involved in different parts of the complement system and metabolic and inflammatory processes, e.g., complement factor B, vitamin D-binding protein, ceruloplasmin, transthyretin and alpha-2-HS-glycoprotein. These results indicate that important systemic protein differences exist between women with CWP and healthy women. Further, this study illustrates the potential use of proteomics to detect biomarkers that may provide new insights into the molecular mechanism(s) of chronic pain. However, further larger investigations are required in order to confirm these findings before it will be possible to identify proteins as potential pain biomarkers for clinical use.


The ossification centers in the lower end of tibiotarsus in domestic fowl.
The morphogenesis of lower end of tibia in chick is studied commonly but the process of ossification of the same has received very little attention so far. The present study is directed to throw some light on the appearance of ossification centers in the lower end of tibiotarsus of chick. The histology of lower end of tibiotarsus was studied by procuring developing tibiotarsi from chick embryos (20) of 6th day incubation till hatching and 3 post hatched chicks. The transparancies of chick embryos at different incubation periods and post hatched chicks were prepared by Dawson's Alizarin staining method. Three cartilage center (tibial, fibulare and intermedium) appeared in 6...9 days of incubation period in the tarsal region. These gradually fused with the lower end of tibia. Three ossification centres developed in the lower end of tibiotarsus. One for intermedium appeared on 16th day and two fotibial and fibulare on 20th day. All these three centres could be located in the transparancies of the chick embryos in tarsal region. The present study proves that the three cartilages centres maintain their individuality during the ossification process even though those fuse completely with the lower end of tibia in chick. The centers for tibial and fibulare are similar to epiphyseal centres of mammals in histological details.


Patterns of morphological changes and hybridization between sympatric whitefish morphs (Coregonus spp.) in a Swiss lake: a role for eutrophication?
Whitefish, genus Coregonus, show exceptional levels of phenotypic diversity with sympatric morphs occurring in numerous postglacial lakes in the northern hemisphere. Here, we studied the effects of human-induced eutrophication on sympatric whitefish morphs in the Swiss lake, Lake Thun. In particular, we addressed the questions whether eutrophication (i) induced hybridization between two ecologically divergent summer-spawning morphs through a loss of environmental heterogeneity, and (ii) induced rapid adaptive morphological changes through changes in the food web structure. Genetic analysis based on 11 microsatellite loci of 282 spawners revealed that the pelagic and the benthic morph represent highly distinct gene pools occurring at different relative proportions on all seven known spawning sites. Gill raker counts, a highly heritable trait, showed nearly discrete distributions for the two morphs. Multilocus genotypes characteristic of the pelagic morph had more gill rakers than genotypes characteristic of benthic morph. Using Bayesian methods, we found indications of recent but limited introgressive hybridization. Comparisons with historical gill raker data yielded median evolutionary rates of 0.24 haldanes and median selection intensities of 0.27 for this trait in both morphs for 1948-2004 suggesting rapid evolution through directional selection at this trait. However, phenotypic plasticity as an alternative explanation for this phenotypic change cannot be discarded. We hypothesize that both the temporal shifts in mean gill raker counts and the recent hybridization reflect responses to changes in the trophic state of the lake induced by pollution in the 1960s, which created novel selection pressures with respect to feeding niches and spawning site preferences.


No prosthetic management of massive and irreparable rotator cuff tears.
A massive rotator cuff tear is not necessarily irreparable. Number of tendons involved, muscle-tendon unit quality, and decreased acromionhumeral distance (AHD) are as important as tear size in determining reparability of lesion. Massive and irreparable rotator cuff tears cannot be anatomically repaired to the bone and are a common source of pain and disability even in middle-aged patients. In these patients when conservative management has failed, it is possible to perform different surgical techniques. A functional repair can help to restore the horizontal force couple of the cuff on the humeral head and to increase the AHD. Debridement of irreparable tears and biceps tenotomy or tenodesis can have a role in low functional demand patients but results deteriorate over time. Recently, several commercially available tissue-engineered biological and synthetic scaffolds have been developed to augment rotator cuff repairs. The aim is to provide a mechanical improvement in case of poor quality tissue at time zero and give a support to have a better cuff healing. In selected cases, the scaffold can be used also to bridge tendon defect. Patients who not have pseudoparalysis, cuff tear arthropathy and with intact deltoid function can benefit from tendon transfers with satisfactory outcomes. These different procedures should be chosen for each patient with selected criteria and after a satisfactory explanation about the really possible expectation after surgery.


van der Waals interactions between nanotubes and nanoparticles for controlled assembly of composite nanostructures.
We have demonstrated that ubiquitous van der Waals forces are significant in controlling the interactions between nanoparticles and nanotubes. The adsorption of gold nanoparticles (AuNPs) on nanotubes (MWNTs) obeys a simple quadratic dependence on the nanotube surface area, regardless of the source of AuNPs and MWNTs. Changes in the geometric parameters of the components have pronounced effects on the affinity of nanoparticles for nanotubes, with larger, more polarizable nanostructures exhibiting stronger attractive interactions, the impact of which changes in the following order MWNT diameter > AuNP diameter > MWNT length.


Starting antidepressant use: a qualitative synthesis of UK and Australian data.
To explore people's experiences of starting antidepressant treatment. Qualitative interpretive approach combining thematic analysis with constant comparison. Relevant coding reports from the original studies (generated using NVivo) relating to initial experiences of antidepressants were explored in further detail, focusing on the ways in which participants discussed their experiences of taking or being prescribed an antidepressant for the first time. 108 men and women aged 22-84 who had taken antidepressants for depression. Respondents recruited throughout the UK during 2003-2004 and 2008 and 2012-2013 and in Australia during 2010-2011. People expressed a wide range of feelings about initiating antidepressant use. People's attitudes towards starting antidepressant use were shaped by stereotypes and stigmas related to perceived drug dependency and potentially extreme side effects. Anxieties were expressed about starting use, and about how long the antidepressant might begin to take effect, how much it might help or hinder them, and about what to expect in the initial weeks. People worried about the possibility of experiencing adverse effects and implications for their senses of self. Where people felt they had not been given sufficient time during their consultation information or support to take the medicines, the uncertainty could be particularly unsettling and impact on their ongoing views on and use of antidepressants as a viable treatment option. Our paper is the first to explore in-depth patient existential concerns about start of antidepressant use using multicountry data. People need additional support when they make decisions about starting antidepressants. Health professionals can use our findings to better understand and explore with patients' their concerns before their patients start antidepressants. These insights are key to supporting patients, many of whom feel intimidated by the prospect of taking antidepressants, especially during the uncertain first few weeks of treatment.


[Modern approaches to diagnostics and treatment of chronic pancreatitis].
This review expounds modern views of diagnostic and treatment of chronic hepatitis (CH). Diagnosis of CH is possible only by precise methods of non-invasive morphological study, such as multispiral CT and/or endoscopic ultrasonography. Severe pain syndrome in CH requires the use of analgesics, pancreatin, tricyclic antidepressants, and anti-oxidant vitamin-mineral complexes. Low effectiveness of such therapy is an indication for surgical and endoscopic treatment.


Alcohol and the liver.
To apprise the reader of advances in 2005 in the epidemiology, pathogenesis, prognosis and treatment of alcoholic liver disease. Alcohol use has declined in developed countries, but the opposite is true elsewhere; alcoholic liver disease is a considerable burden worldwide. Genetic mechanisms for alcoholic liver disease are being discovered in addition to aggravating cofactors, such as hepatitis C, obesity and iron overload, and ameliorating ones, like coffee and tea drinking. The involvement of the innate immune system and the mechanisms of apoptosis in alcoholic liver disease are better appreciated, especially the emerging role of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Steroid use and nutrition for alcoholic hepatitis are being refined, and the validity of the model for end-stage liver disease (MELD) score in predicting the outcome of alcoholic liver disease is upheld. Recidivism after liver transplantation for alcoholic liver disease adversely impacts long-term survival. Inroads are being made into the genetics of alcoholic liver disease and new phenomena are being uncovered in its pathogenesis, but safe and effective therapies for both alcoholic hepatitis and alcoholic cirrhosis are still wanting.


Influence of rearing temperature and feed format in the development of the pendulous crop in broilers.
The pendulous crop is characterized by excessive distension of the crop musculature, compromising the bird's productivity and welfare. The etiology is still unknown, but it is believed that factors related to the birds' handling might be related to its incidence. The study was conducted in 2 environmental chambers. One was maintained at a comfortable temperature, while the other was set at a much lower temperature. In each chamber, animals were divided into 16 experimental pens (8 received mash feed and the others received pelletized feed) with a density of 12 birds/m2 (an expected stocking density of 32-36 kg/m2 after 42 d). The effects of rearing temperatures were evaluated in terms of broiler performance, specifically weight gain (kg), feed intake (kg), weekly feed intake (kg/wk), and feed conversion (kgfeed/kggrowth). The occurrences of pendulous crop were quantified every 2 d after the 14th day of rearing. Birds grown in thermal comfort and fed a pelletized ration were most susceptible (12%) to pendulous crop, followed by birds fed pelletized feed and reared in cold conditions (6.8%), and birds given mashed feed and reared at either temperature (about 3%). We concluded that feeding pelleted feed combined with warmer rearing temperatures may have caused some alteration of the gastrointestinal system of birds, which caused pendulous crop to be more prevalent.


Inhibition failures and late errors in the antisaccade task: influence of cue delay.
In the antisaccade task participants are required to saccade in the opposite direction of a peripheral visual cue (PVC). This paradigm is often used to investigate inhibition of reflexive responses as well as voluntary response generation. However, it is not clear to what extent different versions of this task probe the same underlying processes. Here, we explored with the Stochastic Early Reaction, Inhibition, and late Action (SERIA) model how the delay between task cue and PVC affects reaction time (RT) and error rate (ER) when pro- and antisaccade trials are randomly interleaved. Specifically, we contrasted a condition in which the task cue was presented before the PVC with a condition in which the PVC served also as task cue. Summary statistics indicate that ERs and RTs are reduced and contextual effects largely removed when the task is signaled before the PVC appears. The SERIA model accounts for RT and ER in both conditions and better so than other candidate models. Modeling demonstrates that voluntary pro- and antisaccades are frequent in both conditions. Moreover, early task cue presentation results in better control of reflexive saccades, leading to fewer fast antisaccade errors and more rapid correct prosaccades. Finally, high-latency errors are shown to be prevalent in both conditions. In summary, SERIA provides an explanation for the differences in the delayed and nondelayed antisaccade task. NEW & NOTEWORTHY In this article, we use a computational model to study the mixed antisaccade task. We contrast two conditions in which the task cue is presented either before or concurrently with the saccadic target. Modeling provides a highly accurate account of participants' behavior and demonstrates that a significant number of prosaccades are voluntary actions. Moreover, we provide a detailed quantitative analysis of the types of error that occur in pro- and antisaccade trials.


High hydroxyurea usage in sickle cell anemia regardless of patient demographics.
Hydroxyurea is highly effective in sickle cell disease, but it is still underutilized. Reports of hydroxyurea utilization largely use Medicaid data, and socioeconomics is often cited as a barrier. To address whether patient demographics influenced the high hydroxyurea usage rate recently reported for the pediatric sickle cell program of Northern Virginia, analysis of data from 2011 to 2021 revealed no statistical difference in hydroxyurea usage rate between Medicaid and non-Medicaid, African American and African, or age less than 13 and age greater than or equal to 13 years cohorts, demonstrating that hydroxyurea can be successfully implemented across demographic groups.


Ochratoxin A in Moroccan foods: occurrence and legislation.
Ochratoxin A (OTA) is secondary metabolite naturally produced in food and feed by toxigenic fungi, especially some Aspergillus species and Penicillium verucosum. OTA is one of the most studied mycotoxins and is of great interest due to its toxic effects on human and animals. OTA is produced in different food and feed matrices and contaminates a large range of base foods including cereals and derivatives, spices, dried fruits, wine and coffee, etc. Morocco, a North African country, has a climate characterized by high humidity and temperature, which probably favors the growth of molds. This contribution gives an overview of principal investigations about the presence of OTA in foods available in Morocco. Due to its toxicity, OTA presence is increasingly regulated worldwide, especially in countries of the European Union. However, up until now, no regulation limits were in force in Morocco, probably due to the ignorance of the health and economic problems resulting from OTA contamination. Finally, recommendations and future research directions are given required to assess the situation completely.


Does Extrathoracic Jugulo-Femoral Venous Shunt Avoid Cardiopulmonary Bypass in Glenn Bi-Directional Shunt Procedure?
The aim of the present study was to evaluate the efficacy of extrathoracic jugulo-femoral venous shunt (JFVS) in avoiding the need for cardiopulmonary bypass (CPB) during the bi-directional Glenn shunt (BDGS) procedure. A total of 40 patients aged between 9 and 36 months were enrolled in the present study. The patients were classified into two groups, group A (n=20) in which the patients had no veno-venous shunt and group B (n=20) in which the patients had extrathoracic JFVS. Patients requiring CPB, arterial oxygen saturation (SaO2), heart rate, mean arterial pressure (MAP) and central venous pressure (CVP) were recorded during surgery. Postoperative time of intubation, intensive care unit (ICU) and hospital length of stays and neurological complications were also recorded. The number of patients who needed urgent CPB was greater in group A than in group B. Intraoperative MAP was significantly lower in group A than in group B 10 min after clamping of the superior vena cava (SVC) and 30 min after declamping of the SVC. The CVP was significantly lower, and arterial SaO2 was significantly higher in group B than in group A 10 min after clamping of the SVC. The duration of postoperative intubation was significantly shorter in group B than in group A, and the ICU length of stay was shorter in group B than in group A. The hospital length of stay was similar in both groups. Postoperative neurological deficits were comparable in both groups. The use of extrathoracic JFVS during the BDGS procedure avoided the use of CPB, maintained MAP and prevented any significant increase in SVC pressure.


Phonology, reading acquisition, and dyslexia: insights from connectionist models.
The development of reading skill and bases of developmental dyslexia were explored using connectionist models. Four issues were examined: the acquisition of phonological knowledge prior to reading, how this knowledge facilitates learning to read, phonological and nonphonological bases of dyslexia, and effects of literacy on phonological representation. Compared with simple feedforward networks, representing phonological knowledge in an attractor network yielded improved learning and generalization. Phonological and surface forms of developmental dyslexia, which are usually attributed to impairments in distinct lexical and nonlexical processing "routes," were derived from different types of damage to the network. The results provide a computationally explicit account of many aspects of reading acquisition using connectionist principles.


The application of topical anesthesia combined with subconjunctival anesthesia for glaucoma surgery.
To evaluate the feasibility, reliability and analgesia effect of topical anesthesia combined with subconjunctival anesthesia in anti-glaucomatous surgery. Two hundred and four cases (357 eyes) underwent anti-glaucomatous surgeries under topical anesthesia with 0.5% Alcaine eye drops combined with subconjunctival anesthesia with 2% Lidocaine. The analgesic effect was analysed with visual analogue pain scale. Among all of 357 eyes, 62 eyes underwent peripheral iridectomy, 67 eyes underwent simple trabeculectomy, 167 eyes underwent compound brabeculectomy and 12 eyes nonpenetrating trabecular surgery. The effects of anesthesia were as follows: 304 eyes (85.2%) were painless (Grade I), 50 eyes (14.0%) were slight painful (Grade II), and 3 eyes (0.8%) were more painful (Grade III) during surgery. And no severe complications were observed in all the cases during surgery and postoperatively. Amaurosis fugax was not observed in the glaucoma patients at the late stage with narrow visual fields and poor visual ability. Topical anesthesia combined with subconjunctival anesthesia is effective, safe and simple anesthesia alternative in routine anti-glaucomatous surgery, especially for the glaucoma patients at the late stage with narrow visual fields and poor visual ability. It is worthy of being applied widely.


A protein disulfide isomerase/thioredoxin-1 complex is physically attached to exofacial membrane tumor necrosis factor receptors: overexpression in chronic lymphocytic leukemia cells.
The 3D structures and functions of cysteine-rich receptors such as tumor necrosis factor receptors (TNFRs) are redox-modulated by dithiol-disulfide exchange. TNFR superfamily members participate in growth regulation in B-cell chronic lymphocytic leukemia (CLL), and tissue stromal cells interact with leukemia cells, profoundly affecting their viability via release of redox-active components, including cysteine, thioredoxin-1 (Trx1), and Trx reductase. Trx1 was previously shown to enhance release of TNF, which acts as an autocrine/paracrine growth factor in CLL. The nature of the mechanism is not known, however. Here, we investigated whether Trx1 and protein disulfide isomerase (PDI), a chaperone and Trx-family member, may interact with TNFRs. We found direct physical association between PDI and TNFR1 or TNFR2 by coclustering and affinity isolation. PDI (57 kDa) formed covalent/reduction-sensitive 69-kDa complexes with Trx1 (12 kDa) in a majority of CLL cell samples, detected at low levels only in control B-cells. Functionally, the TNF/TNFR signaling via the nuclear factor kappa B-driven autocrine loop was disrupted in a dose-dependent fashion by PDI-inhibitors bacitracin, anti-PDI, or anti-Trx1 antibodies, resulting in reduced viability. PDI was significantly overexpressed in immunoglobulin heavy-chain variable (IGHV) unmutated versus mutated CLL (p=0.0102), and amplified TNF release was observed in the former group. This study points out a previously unrecognized physical and functional association of TNFRs with the redox-active proteins PDI and Trx1. We describe here a new level of TNF regulation, in which membrane TNFRs are redox controlled at the exofacial surface by PDI/Trx1. These findings shed new light on the observed survival benefit in CLL B-cells exerted by TNFR-superfamily ligands and point at potential therapeutic strategies.


Production of monoclonal antibodies to the high fibrin-affinity, tissue-type plasminogen activator of human plasma. Demonstration of its endothelial origin by immunolocalization.
Monoclonal antibodies (MAbs) to vascular plasminogen activator (vPA), the tissue-type plasminogen activator (tPA) in human plasma, were produced to be used as probes for immunochemical analysis. Human tissue sections and one of these MAbs were used to demonstrate the endothelial origin of plasma-tPA by immunohistochemistry. To produce MAbs, mice were immunized with semipurified vPA isolated from postocclusion human venous blood. Primed spleen cells were fused with the mouse myeloma cell line NS-1. Screening for MAb-producing hybridomas was performed with postocclusion euglobulins as a source of antigen by means of a solid-phase fibrin-vPA immunoassay. The selective and high-affinity binding of vPA for fibrin ensures the specificity and sensitivity of this test. Thus, eight hybridomas secreting MAbs to vPA were selected, cloned, and established as permanent hybridoma cell lines. Immunohistochemical analysis of cryostat sections of human tissues was performed with EA-delta 12D, a MAb having no inhibitory effect against vPA activity but binding to vPA with a high affinity. Thus, the only structures immunostained were endothelial cells of venules, capillaries, and arterioles. The EA-delta 12D monoclonal localization of plasma vPA in the endothelial lining of blood vessels provides evidence that tPA in plasma originates from the vascular wall and validates its designation as vascular plasminogen activator, ie, vPA. Also, our results are consistent with the fact that vPA in blood and tPA in tissues are immunologically identical and have a common endothelial origin.


Fast chemical force microscopy demonstrates that glycopeptidolipids define nanodomains of varying hydrophobicity on mycobacteria.
Mycobacterium abscessus is an emerging multidrug-resistant bacterial pathogen causing severe lung infections in cystic fibrosis patients. A remarkable trait of this mycobacterial species is its ability to form morphologically smooth (S) and rough (R) colonies. The S-to-R transition is caused by the loss of glycopeptidolipids (GPLs) in the outer layer of the cell envelope and correlates with an increase in cording and virulence. Despite the physiological and medical importance of this morphological transition, whether it involves changes in cell surface properties remains unknown. Herein, we combine recently developed quantitative imaging (QI) atomic force microscopy (AFM) with hydrophobic tips to quantitatively map the surface structure and hydrophobicity of M. abscessus at high spatiotemporal resolution, and to assess how these properties are modulated by the S-to-R transition and by treatment with an inhibitor of the mycolic acid transporter MmpL3. We discover that loss of GPLs leads to major modifications in surface hydrophobicity, without any apparent change in cell surface ultrastructure. While R bacilli are homogeneously hydrophobic, S bacilli feature unusual variations of nanoscale hydrophobic properties. These previously undescribed cell surface nanodomains are likely to play critical roles in bacterial adhesion, aggregation, phenotypic heterogeneity and transmission, and in turn in virulence and pathogenicity. Our study also suggests that MmpL3 inhibitors show promise in nanomedicine as chemotherapeutic agents to interfere with the highly hydrophobic nature of the mycobacterial cell wall. The advantages of QI-AFM with hydrophobic tips are the ability to map chemical and structural properties simultaneously and at high resolution, applicable to a wide range of biosystems.


Preventing Unequal Health Outcomes in COVID-19: A Systematic Review of Past Interventions.
Background: We sought to identify interventions that reduced disparities in health outcomes in infectious disease outbreaks or natural disasters in the United States to understand whether these interventions could reduce health disparities in the current COVID-19 pandemic. Methods: We searched MEDLINE and other databases to May 2020 to find studies that examined interventions to mitigate health inequalities in previous infectious disease pandemics or disasters. We assessed study quality using the Newcastle-Ottawa Scale and the Critical Appraisal Skills Program (CASP) Checklist for Qualitative Studies. Results: We included 14 articles (12 studies) and 5 Centers for Disease Control (CDC) stakeholder meeting articles on pandemic influenza preparedness in marginalized populations. Studies called for intervention and engagement before pandemic or disaster onset. Several studies included interventions that could be adapted to COVID-19, including harnessing technology to reach disadvantaged populations, partnering with trusted community liaisons to deliver important messaging around disease mitigation, and using culturally specific communication methods and messages to best reach marginalized groups. Discussion: To our knowledge this is the first systematic review to examine interventions to mitigate health inequities during an infectious disease pandemic. However, given that we identified very few disparities-focused infectious disease intervention studies, we also included studies from the disaster response literature, which may not be as generalizable to the current context of COVID-19. Overall, community outreach and tailored communication are essential in disease mitigation. More research is needed to evaluate systemic interventions that target the distal determinants of poor health outcomes among marginalized populations during pandemics and natural disasters.


Next generation sequencing-based mutation screening of 86 patients with idiopathic short stature.
Although mutations in ACAN, FGFR3, NPR2, and SHOX typically lead to skeletal dysplasia, and mutations in GHRHR, GH1, GHR, STAT5B, IGF1, IGFALS, and IGF1R usually underlie hormonal defects of the growth hormone (GH)-insulin-like growth factor 1 (IGF1) axis, such mutations have also been identified in patients with idiopathic short stature (ISS). Of these, SHOX abnormalities are known to account for a certain percentage of ISS cases, whereas the frequency of mutations in the other 10 genes in ISS cohorts remains unknown. Here, we performed next-generation sequencing-based mutation screening of the 10 genes in 86 unrelated Japanese ISS patients without SHOX abnormalities. We searched for rare protein-altering variants. The functional significance of the identified variants was assessed by in silico analyses. Consequently, we identified 18 heterozygous rare variants in 19 patients, including four probable damaging variants in ACAN, six pathogenicity-unknown variants in FGFR3, GHRHR, GHR, and IGFALS, and eight possible benign variants. Pathogenic variants in NPR2, GH1, and IGF1 were absent from our cohort. Unlike previously reported patients with ACAN mutations, our four patients with ACAN variants manifested non-specific short stature with age-appropriate or mildly delayed bone ages, and had parents of normal stature. These results indicate that ACAN mutations can underlie ISS without characteristic skeletal features, and that such mutations are possibly associated with de novo occurrence or low penetrance. In addition, our data imply that mutations in FGFR3, NPR2, and GH-IGF1 axis genes play only limited roles in the etiology of ISS.


Mid-term implant survival, functional and radiological results and mechanical complications of mega-prosthetic reconstruction around the knee with the PENTA® system.
Mega-prosthetic reconstruction is the most common treatment method for massive osteoarticular defects caused by tumor resection around the knee. The new implant is a highly modular rotational-hinged megaprosthesis system with a distinct pentagonal stem geometry and variable implantation options. The aim of this study is to present the mid-term implant survival characteristics, functional and radiological results and mechanical complication profile of the new megaprosthesis. One hundred and one mega-prosthetic knee reconstruction procedures in 90 patients (M/F: 51/39) utilizing the new implant system were retrospectively analyzed. In 68 patients, the megaprosthesis was used for primary reconstruction following tumor resection while it was used for revision of other implants in 22. The mean age was 28.5 (7-66) years and the mean follow-up was 59.2 (24-124) months. The most common primary pathology was osteosarcoma with 63-70% patients, the most common anatomical site of involvement was the distal femur with 56-62% patients. Henderson Type 2 failure (aseptic loosening) was seen in only 2-2.2% patients while Type 3 (structural failure) was seen in 29-32.2% Although the 5-year anchorage survival rate was 94.3%, overall mechanical implant survival was 76.1% at 5 years due to a relatively high failure rate in the first-generation hinge mechanism of the implant. The 5-year hinge survival rate demonstrated a significant improvement rate from 61.7% to 87.2% between the first and second generations of the implant (p = 0.027). The mean MSTS score was 24 out of 30 (14-29). The mean cumulative ISOLS radiographic score for index megaprosthesis operations was 19.7 (12-24), which corresponded to excellent outcome. The new megaprosthesis system is a reliable choice for the reconstruction of tumor-related massive osteoarticular defects around the knee. Although long-term follow-up is necessary for a definitive evaluation of the implant's survival characteristics, midterm follow-up yields exceptional anchorage properties related to pentagonal stem geometry with very good functional outcomes.


Error-Correcting Factorization.
Error Correcting Output Codes (ECOC) is a successful technique in multi-class classification, which is a core problem in Pattern Recognition and Machine Learning. A major advantage of ECOC over other methods is that the multi-class problem is decoupled into a set of binary problems that are solved independently. However, literature defines a general error-correcting capability for ECOCs without analyzing how it distributes among classes, hindering a deeper analysis of pair-wise error-correction. To address these limitations this paper proposes an Error-Correcting Factorization (ECF) method. Our contribution is three fold: (I) We propose a novel representation of the error-correction capability, called the design matrix, that enables us to build an ECOC on the basis of allocating correction to pairs of classes. (II) We derive the optimal code length of an ECOC using rank properties of the design matrix. (III) ECF is formulated as a discrete optimization problem, and a relaxed solution is found using an efficient constrained block coordinate descent approach. (IV) Enabled by the flexibility introduced with the design matrix we propose to allocate the error-correction on classes that are prone to confusion. Experimental results in several databases show that when allocating the error-correction to confusable classes ECF outperforms state-of-the-art approaches.


Factor VIII as a marker of endothelial cells in follicular carcinoma of the thyroid.
Factor VIII-related antigen is a recognised marker of endothelial cells. A brief immunocytochemical study of its distribution in follicular carcinoma of the thyroid and its value in the recognition of vascular invasion by this tumour has been carried out. Ten cases of follicular carcinoma of the thyroid were studied. In each strong endothelial staining was found in the majority of vessels in the adjacent normal thyroid. Lymphatic endothelium was negative. In eight of the 10 cases the staining of vessels within the tumour was absent or very weak. Staining was also absent in the majority of vessels completely occluded by tumour, but was present in the endothelium of vessels only partly occluded by tumour. It is concluded that factor VIII-related antigen staining has only limited value in the recognition of vascular invasion in follicular carcinoma. The absence of vascular staining in the tumour leads us to suggest that inhibition of factor VIII production by the tumour could be a possible mechanism which facilitates vascular invasion and metastasis.


Observational study on safety and tolerability of duloxetine in the treatment of female stress urinary incontinence in German routine practice.
To evaluate the safety and tolerability of duloxetine during routine clinical care in women with stress urinary incontinence (SUI) in Germany, and in particular, to identify previously unrecognized safety issues as uncommon adverse reactions, and the influence of confounding factors present in clinical practice on the safety profile of duloxetine. Office-based urologists, gynaecologists and primary care physicians were asked to document women newly started on treatment for moderate to severe symptoms of SUI. Six thousand eight hundred and fifty-four patients from urologist/gynaecologist practices and 5879 primary care patients were assessed. In a two-armed, observational study with parallel 12 week (urologists and gynaecologists) or 24 week (primary care physicians) design, patients were treated with duloxetine or other conservative treatment. The main outcome measure was the occurrence of adverse events (AEs). Baseline characteristics differed slightly between patient groups and studies. Duloxetine doses in most patients were lower than recommended. Overall, AE frequency with duloxetine was lower than in controlled studies (15.9% (95% CI 14.9, 16.9) and 9.1% (95% CI 8.2, 10.0) in the 12 and 24 week treatment groups, respectively), but exhibited a similar qualitative spectrum. In the logistic regression models, the following factors were associated with greater AE risk: investigator specialization (gynaecologist vs. urologist and primary care physician), initial duloxetine dose (80 vs. 20 mg day(-1) ) and use of any concomitant medication. Within the 24 week study, a positive screen for depressive disorder was surprisingly common, but no case of attempted suicide was reported in either study. Our results from German clinical practice show that women with SUI were often treated with duloxetine doses lower than recommended. This was associated with a low incidence of AEs. Suicide attempts were not reported.


Pre-pregnancy endothelial dysfunction and birth outcomes: The Coronary Artery Risk Development in Young Adults (CARDIA) Study.
Endothelial dysfunction is a form of subclinical cardiovascular disease that may be involved in preterm birth and small-for-gestational-age deliveries. However, concentrations of biomarkers of endothelial dysfunction before pregnancy have rarely been measured. We hypothesized that higher levels of biomarkers of endothelial dysfunction (cellular adhesion molecules and selectins) would be associated with odds of preterm birth and/or small-for-gestational-age deliveries. We included 235 women from the Coronary Artery Risk Development in Young Adults (CARDIA) study who were nulliparous at Y7, reported ≥1 live birth through Y25, and had ≥1 biomarker measured at Y7. We tested for associations between individual biomarkers and an averaged z-score representing total endothelial dysfunction with preterm birth and/or small-for-gestational-age deliveries using Poisson regression, adjusted for demographic and clinical characteristics at the exam immediately preceding index birth. At Y7, total evidence of endothelial dysfunction was similar in women who did (n = 59) and did not have (n = 176) preterm birth and/or small-for-gestational-age deliveries. There was no association between biomarkers of endothelial dysfunction (either individual biomarker or total score) with odds of preterm birth and/or small-for-gestational-age deliveries after adjustment: IRR = 1.01, 95% CI: 0.74, 1.39, p = 0.93 for total endothelial biomarker score. Associations were not modified by race. We conclude that biomarkers of endothelial dysfunction in nulliparous women, measured ~3 years before pregnancy, did not identify women at risk for preterm birth and/or small-for-gestational-age deliveries. This suggests that the maternal endothelial dysfunction that is believed to contribute to these birth outcomes may not be detectable before pregnancy.


Results of Salmonella enterica subsp. enterica serotype identification by Salmonella Check&Trace microarray in international External Quality Assurance Systems.
Traditionally Salmonella enterica subsp. enterica serotypes are identified by slide agglutination with specific antisera for somatic, flagellar and sometimes capsular antigens. An alternative way is genoserotyping using for example a microarray, eg. commercially available test Check&Trace Salmonella. The goal of this study was to evaluate the Check&Trace Salmonella microarray for Salmonella enterica subsp. enterica serotype identification, using Salmonella strains provided by reference laboratories during External Quality Assurance Systems organized for national reference laboratories by ECDC and WHO GFN. 80 Salmonella enterica subsp. enterica have been tested using Check & Trace Salmonella (Check-Points BC, Netherlands). Also classical slide agglutination was performed according to EN ISO 6579:2003/Al:2007 norm, used as reference method. In the group of 80 tested strains, 66% were identified correctly, 4% gave uncertain results and 29% showed "Salmonella, genovar" without a serotype, of which 69% were not included in the CTS list of serotypes. Finally one strain has been recognized incorrectly. Because of IVD certification lack, the CTS test could not be recommended to clinical laboratories. AOAC-RI and OIE certification for test cause, that CTS could be used in most food, environmental and veterinary laboratories with the condition, that all unrecognized strains should be sent to a reference laboratory, to type according to EN ISO 6579:2003/Al:2007 norm, by KWM serotyping or other equal alternative methods.


Semisynthesis of Functional Glycosylphosphatidylinositol-Anchored Proteins.
Glypiation is a common posttranslational modification of eukaryotic proteins involving the attachment of a glycosylphosphatidylinositol (GPI) glycolipid. GPIs contain a conserved phosphoglycan that is modified in a cell- and tissue-specific manner. GPI complexity suggests roles in biological processes and effects on the attached protein, but the difficulties to get homogeneous material have hindered studies. We disclose a one-pot intein-mediated ligation (OPL) to obtain GPI-anchored proteins. The strategy enables the glypiation of folded and denatured proteins with a natural linkage to the glycolipid. Using the strategy, glypiated eGFP, Thy1, and the Plasmodium berghei protein MSP119 were prepared. Glypiation did not alter the structure of eGFP and MSP119 proteins in solution, but it induced a strong pro-inflammatory response in vitro. The strategy provides access to glypiated proteins to elucidate the activity of this modification and for use as vaccine candidates against parasitic infections.


Nutrition, Immune Function, and Infectious Disease in Military Personnel: A Narrative Review.
Consuming a diet that meets energy demands and provides essential nutrients promotes a healthy immune system, while both under- and over-nutrition have been associated with immune dysfunction. Military personnel comprise a unique population who frequently endure multi-stressor environments, predisposing them to immune decrements. Additionally, 49% and 22% of active duty U.S. military personnel are classified as overweight and obese, respectively. A literature search on PubMed was conducted to identify studies, reports, review papers, and references within those sources relevant to the topic area. Military personnel experiencing either under- or over-nutrition can suffer from degraded health, readiness, and performance. Insufficient intake of nutrients during military operations increases infection risk and negatively impacts infection recovery. Energy, protein, iron, zinc, and vitamins C and D are nutritional areas of concern that may impact immune competence in a multi-stressor environment. Over-nutrition can promote accretion of excess body fat and obesity, which contributes to a chronic inflammatory state that coincides with immune impairments. Prioritizing efforts to optimize nutrient intake is one approach for reducing disease burden and improving readiness. This review discusses nutritional concerns concomitant to multi-stressor environments that impact immune function, and the relevance of obesity to infectious disease risk in the military population.


Genomic Profiles and Current Therapeutic Agents in Neuroendocrine Neoplasms.
Neuroendocrine neoplasms (NENs) are rare tumors that mainly occur in the gastroenteropancreatic (GEP) tract and lungs. According to the current World Health Organization classification for GEP-NENs and lung NENs, treatment strategies differ for well-differentiated and poorly differentiated subtypes. For well-differentiated GEP-NENs, somatostatin analogues (SSA), peptide receptor radionuclide therapy, and molecular-targeted agents are approved as the standards of care based on phase III clinical trial data. Promising data regarding the use of everolimus and the novel SSA pasireotide for lung NENs are emerging, though additional studies are required to confirm these effects. For poorly differentiated tumors from the GEP tract and lung, a platinum-based cytotoxic regimen is widely used. Genomic analysis has recently revealed a diverse pattern of primary organ-dependent mutations, and the use of traditional treatment strategies versus organ-specific strategies is currently under discussion. In addition, clinical trials for several molecular-targeted agents and immune checkpoint inhibitors for the treatment of NENs are currently underway. Accumulating genomic information is expected to contribute to the development of novel therapies for other organ-derived NENs or poorly differentiated neuroendocrine carcinomas (NECs). Here, we provide an updated overview of the current knowledge regarding genomic profiles and representative agents for NENs and highlight the prospects for future investigations.


Behavioral Tests for Associative Learning in Caenorhabditis elegans.
Learning is critical for survival as it provides the capacity to adapt to a changing environment. At the molecular and cellular level, learning leads to alterations within neural circuits that include synaptic rewiring, synaptic plasticity, and protein level/gene expression changes. There has been substantial progress in recent years on dissecting how learning and memory is regulated at the molecular and cellular level, including the use of compact invertebrate nervous systems as experimental models. This progress has been facilitated by the establishment of robust behavioral assays that generate a quantifiable readout of the extent to which animals learn and remember. This chapter will focus on protocols of behavioral tests for associative learning using the nematode Caenorhabditis elegans, with its unparalleled genetic tractability, compact nervous system of ~300 neurons, high level of conservation with mammalian systems, and amenability to a suite of behavioral tools and analyses. Specifically, we will provide a detailed description of the methods for two behavioral assays that model associative learning, one measuring appetitive olfactory learning and the other assaying aversive gustatory learning.


The role of sodium pump activity in the hyperpolarization and in subsequent depolarization of smooth muscle in response to stimulation of post-synaptic alpha 1-adrenoceptors.
The electrical and mechanical activities of guinea pig taenia coli smooth muscle were measured by a "sucrose gap" technique. Under the same experimental conditions the ionic content of smooth muscle was also measured. The mean value of the resting potential was 56.9 +/- 1.1 mV (S.E.M.; n = 46). In normal Krebs solution immediately after dissection intracellular sodium amounted to 30.1, and intracellular calcium to 1.5 mmole x kg-1 wet weight. In response to adrenaline administration there was a Ca-dependent hyperpolarization (peak, 6.8 +/- 0.3 mV S.E.M.; n = 5) and an increased Na efflux with a rate constant (k) of 0.16 min-1 (60'). Removal of adrenaline was followed by so-called "postadrenaline depolarization" i.e. the decrease of the membrane potential was greater than the initial rise, an effect enhanced by ouabain (2 X 10(-5) M). Clonidine (5.3 X 10(-6) M), a selective presynaptic-adrenoceptor (alpha 2-receptor) stimulant failed to produce hyperpolarization, however, phenylephrine (5 X 10(-5) M) a pure postsynaptic alpha-adrenoceptor (alpha 1-receptor) stimulant produced a similar effect as adrenaline. In addition, yohimbine (1.4 X 10(-6) M), a typical presynaptic alpha-adrenoceptor inhibitor failed to affect the action of adrenaline or phenylephrine. These facts indicate that the alpha 1-adrenoceptors present on the smooth muscle are different from those situated presynaptically on the cholinergic nerve terminals modulating the release of acetylcholine. The effect of ouabain to lower membrane potential proved to be Ca2+-dependent. The intracellular sodium content was enhanced by ouabain from 30.1 to 90.9 +/- 4.7 mmole x kg-1 wet weight (60'). On washing out ouabain, hyperpolarization "post-ouabain hyperpolarization" was detected, i.e. the rise of membrane potential was greater than the initial fall. It is suggested that the sodium pump plays a significant role in the post-ouabain hyperpolarization. Direct calculation of sodium movements suggests that the sodium permeability is about 3.7 p-mole x cm-2 x sec-1 and the pump generates a current of 1.06 X 10(-7) A x cm-2 which, in fact, while crossing the smooth muscle membrane with a resistance of 30-60 k omega cm2 would produce a potential difference of 3.2-6.4 mV. It is suggested that the electrogenic sodium pump may contribute to the resting membrane potential about 15 mV, a value obtained from the summation of depolarization induced by ouabain and hyperpolarization produced by adrenaline.


Meta-analysis of the effects of dietary vitamin E supplementation on α-tocopherol concentration and lipid oxidation in pork.
Meta-analyses have been carried out to quantify the effect of dietary vitamin E on α-tocopherol accumulation and on lipid oxidation in porcine M. longissimus. Published results of 13 (vitamin E accumulation) and 10 (lipid oxidation) experiments respectively were used for the analyses. After a number of standardization procedures, a nonlinear relationship was found between the supplementary vitamin E and the accumulation of α-tocopherol in pork which approached a maximum value of 6.4 μg/g tissue. Pork lipid oxidation levels were described in terms of Thiobarbituric Acid Reacting Substances (TBARS) values. The statistical analysis revealed significant effect of vitamin E dose, muscle α-tocopherol concentration and supplementation time on TBARS, resulting in two prediction models for lipid oxidation. Meta-analysis has proven to be a valuable tool for combining results from previous studies to quantify the effects of dietary vitamin E. Further studies, carried out with standardized experimental protocols would be beneficial for model validation and to increase the predictive power of the derived models.