Circulating tumor cells, disease progression, and survival in metastatic breast cancer.
We tested the hypothesis that the level of circulating tumor cells can predict survival in metastatic breast cancer. In a prospective, multicenter study, we tested 177 patients with measurable metastatic breast cancer for levels of circulating tumor cells both before the patients were to start a new line of treatment and at the first follow-up visit. The progression of the disease or the response to treatment was determined with the use of standard imaging studies at the participating centers. Outcomes were assessed according to levels of circulating tumor cells at baseline, before the patients started a new treatment for metastatic disease. Patients in a training set with levels of circulating tumor cells equal to or higher than 5 per 7.5 ml of whole blood, as compared with the group with fewer than 5 circulating tumor cells per 7.5 ml, had a shorter median progression-free survival (2.7 months vs. 7.0 months, P<0.001) and shorter overall survival (10.1 months vs. >18 months, P<0.001). At the first follow-up visit after the initiation of therapy, this difference between the groups persisted (progression-free survival, 2.1 months vs. 7.0 months; P<0.001; overall survival, 8.2 months vs. >18 months; P<0.001), and the reduced proportion of patients (from 49 percent to 30 percent) in the group with an unfavorable prognosis suggested that there was a benefit from therapy. The multivariate Cox proportional-hazards regression showed that, of all the variables in the statistical model, the levels of circulating tumor cells at baseline and at the first follow-up visit were the most significant predictors of progression-free and overall survival. The number of circulating tumor cells before treatment is an independent predictor of progression-free survival and overall survival in patients with metastatic breast cancer.


The transcriptional activator GCN4 contains multiple activation domains that are critically dependent on hydrophobic amino acids.
GCN4 is a transcriptional activator in the bZIP family that regulates amino acid biosynthetic genes in the yeast Saccharomyces cerevisiae. Previous work suggested that the principal activation domain of GCN4 is a highly acidic segment of approximately 40 amino acids located in the center of the protein. We conducted a mutational analysis of GCN4 with a single-copy allele expressed under the control of the native promoter and translational control elements. Our results indicate that GCN4 contains two activation domains of similar potency that can function independently to promote high-level transcription of the target genes HIS3 and HIS4. One of these domains is coincident with the acidic activation domain defined previously; the other extends over the N-terminal one-third of the protein. Both domains are partially dependent on the coactivator protein ADA2. Each domain appears to be composed of two or more small subdomains that have additive effects on transcription and that can cooperate in different combinations to promote high-level expression of HIS3 and HIS4. At least three of these subdomains are critically dependent on bulky hydrophobic amino acids for their function. Five of the important hydrophobic residues, Phe-97, Phe-98, Met-107, Tyr-110, and Leu-113, fall within a region of proposed sequence homology between GCN4 and the herpesvirus acidic activator VP16. The remaining three residues, Trp-120, Leu-123, and Phe-124, are highly conserved between GCN4 and its Neurospora counterpart, cpc-1. Because of the functional redundancy in the activation domain, mutations at positions 97 and 98 must be combined with mutations at positions 120 to 124 to observe a substantial reduction in activation by full-length GCN4, and substitution of all eight hydrophobic residues was required to inactivate full-length GCN4. These hydrophobic residues may mediate important interactions between GCN4 and one or more of its target proteins in the transcription initiation complex.


Glutamate and depression: clinical and preclinical studies.
The past decade has seen a steady accumulation of evidence supporting a role for the excitatory amino acid (EAA) neurotransmitter, glutamate, and its receptors in depression and antidepressant activity. To date, evidence has emerged indicating that N-methyl-d-aspartate (NMDA) receptor antagonists, group I metabotropic glutamate receptor (mGluR1 and mGluR5) antagonists, as well as positive modulators of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors have antidepressant-like activity in a variety of preclinical models. Moreover, antidepressant-like activity can be produced not only by drugs modulating the glutamatergic synapse, but also by agents that affect subcellular signaling systems linked to EAA receptors (e.g., nitric oxide synthase). In view of the extensive colocalization of EAA and monoamine markers in nuclei such as the locus coeruleus and dorsal raphe, it is likely that an intimate relationship exists between regulation of monoaminergic and EAA neurotransmission and antidepressant effects. Further, there is also evidence implicating disturbances in glutamate metabolism, NMDA, and mGluR1,5 receptors in depression and suicidality. Finally, recent data indicate that a single intravenous dose of an NMDA receptor antagonist is sufficient to produce sustained relief from depressive symptoms. Taken together with the proposed role of neurotrophic factors in the neuroplastic responses to stressors and antidepressant treatments, these findings represent exciting and novel avenues to both understand depressive symptomatology and develop more effective antidepressants.


Effects of environmental variation during seed production on seed dormancy and germination.
The environment during seed production has major impacts on the behaviour of progeny seeds. It can be shown that for annual plants temperature perception over the whole life history of the mother can affect the germination rate of progeny, and instances have been documented where these affects cross whole generations. Here we discuss the current state of knowledge of signal transduction pathways controlling environmental responses during seed production, focusing both on events that take place in the mother plant and those that occur directly as a result of environmental responses in the developing zygote. We show that seed production environment effects are complex, involving overlapping gene networks active independently in fruit, seed coat, and zygotic tissues that can be deconstructed using careful physiology alongside molecular and genetic experiments.


Slowed atrial and atrioventricular conduction and depressed HRV in a murine model of hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is a common heritable cardiac disorder with diverse clinical outcomes including sudden death, heart failure, and stroke. Depressed heart rate variability (HRV), a measure of cardiac autonomic regulation, has been shown to predict mortality in patients with cardiovascular disease. Cardiac autonomic remodelling in animal models of HCM are not well characterised. This study analysed Gly203Ser cardiac troponin-I transgenic (TG) male mice previously demonstrated to develop hallmarks of HCM by age 21 weeks. 33 mice aged 30 and 50 weeks underwent continuous electrocardiogram (ECG) recording for 30 min under anaesthesia. TG mice demonstrated prolonged P-wave duration (P < 0.001) and PR intervals (P < 0.001) compared to controls. Additionally, TG mice demonstrated depressed standard deviation of RR intervals (SDRR; P < 0.01), coefficient of variation of RR intervals (CVRR; P < 0.001) and standard deviation of heart rate (SDHR; P < 0.001) compared to controls. Additionally, total power was significantly reduced in TG mice (P < 0.05). No significant age-related difference in either strain was observed in ECG or HRV parameters. Mice with HCM developed slowed atrial and atrioventricular conduction and depressed HRV. These changes were conserved with increasing age. This finding may be indicative of atrial and ventricular hypertrophy or dysfunction, and perhaps an indication of worse clinical outcome in heart failure progression in HCM patients.


Application of Convolutional Neural Network Algorithms for Advancing Sedentary and Activity Bout Classification.
Machine learning has been used for classification of physical behavior bouts from hip-worn accelerometers; however, this research has been limited due to the challenges of directly observing and coding human behavior "in the wild." Deep learning algorithms, such as convolutional neural networks (CNNs), may offer better representation of data than other machine learning algorithms without the need for engineered features and may be better suited to dealing with free-living data. The purpose of this study was to develop a modeling pipeline for evaluation of a CNN model on a free-living data set and compare CNN inputs and results with the commonly used machine learning random forest and logistic regression algorithms. Twenty-eight free-living women wore an ActiGraph GT3X+accelerometer on their right hip for 7 days. A concurrently worn thigh-mounted activPAL device captured ground truth activity labels. The authors evaluated logistic regression, random forest, and CNN models for classifying sitting, standing, and stepping bouts. The authors also assessed the benefit of performing feature engineering for this task. The CNN classifier performed best (average balanced accuracy for bout classification of sitting, standing, and stepping was 84%) compared with the other methods (56% for logistic regression and 76% for random forest), even without performing any feature engineering. Using the recent advancements in deep neural networks, the authors showed that a CNN model can outperform other methods even without feature engineering. This has important implications for both the model's ability to deal with the complexity of free-living data and its potential transferability to new populations.


ADAP Promotes Degranulation and Migration of NK Cells Primed During in vivo Listeria monocytogenes Infection in Mice.
The adhesion and degranulation-promoting adaptor protein (ADAP) serves as a multifunctional scaffold and is involved in the formation of immune signaling complexes. To date only limited and moreover conflicting data exist regarding the role of ADAP in NK cells. To extend existing knowledge we investigated ADAP-dependency of NK cells in the context of in vivo infection with the intracellular pathogen Listeria monocytogenes (Lm). Ex vivo analysis of infection-primed NK cells revealed impaired cytotoxic capacity in NK cells lacking ADAP as indicated by reduced CD107a surface expression and inefficient perforin production. However, ADAP-deficiency had no global effect on NK cell morphology or intracellular distribution of CD107a-containing vesicles. Proteomic definition of ADAPko and wild type NK cells did not uncover obvious differences in protein composition during the steady state and moreover, similar early response patterns were induced in NK cells upon infection independent of the genotype. In line with protein network analyses that suggested an altered migration phenotype in naïve ADAPko NK cells, in vitro migration assays uncovered significantly reduced migration of both naïve as well as infection-primed ADAPko NK cells compared to wild type NK cells. Notably, this migration defect was associated with a significantly reduced expression of the integrin CD11a on the surface of splenic ADAP-deficient NK cells 1 day post-Lm infection. We propose that ADAP-dependent alterations in integrin expression might account at least in part for the fact that during in vivo infection significantly lower numbers of ADAPko NK cells accumulate in the spleen i.e., the site of infection. In conclusion, we show here that during systemic Lm infection in mice ADAP is essential for efficient cytotoxic capacity and migration of NK cells.


Body-Packing: A Rare Diagnosis to Keep in Mind.
Body packing was first described in 1973 and refers to the intracorporeal concealment of illegal drugs, which are swallowed or placed in anatomical cavities and/or body orifices. The body packer can be asymptomatic or can have signs of systemic drug toxicity (neurological, cardiac, abdominal, renal and cutaneous) due to rupture of the packet(s) or symptoms of gastrointestinal obstruction or perforation. The diagnosis is established based on a suggestive history, findings on physical examination and laboratory findings and/or imaging. The vast majority of patients are asymptomatic and are treated conservatively. However, complex situations may require surgical intervention. We present a case of a 50-year-old man who was admitted in the emergency department with a generalized tonic-clonic seizure and vomiting with plastic film, which raised the suspicion of foreign body ingestion, confirmed by imaging and laboratory tests. He underwent exploratory laparotomy to remove the packages. Body packing is a potentially lethal activity.Body-packers can be asymptomatic, or have signs/symptoms of systemic drug toxicity or gastrointestinal obstruction or perforation.It is essential to recognize this condition so that the correct clinical approach, diagnosis and management can be established.


Animal source foods, rich in essential amino acids, are important for linear growth and development of young children in low- and middle-income countries.
Growth faltering under 5 years of age is unacceptably high worldwide, and even more children, while not stunted, fail to reach their growth potential. The time between conception and 2 years of age is critical for development. The period from 6 to 23 months, when complementary foods are introduced, coincides with a time when growth faltering and delayed neurocognitive developments are most common. Fortunately, this is also the period when diet exercises its greatest influence. Growing up in an adverse environment, with a deficient diet, as typically seen in low- and middle-income countries (LMICs), hampers growth and development of children and prevents them from realising their full developmental and economic future potential. Sufficient nutrient availability and utilisation are paramount to a child's growth and development trajectory, especially in the period after breastfeeding. This review highlights the importance of essential amino acids (EAAs) in early life for linear growth and, likely, neurocognitive development. The paper further discusses signalling through mammalian target of rapamycin complex 1 (mTORC1) as one of the main amino acid (AA)-sensing hubs and the master regulator of both growth and neurocognitive development. Children in LMICs, despite consuming sufficient total protein, do not meet their EAA requirements due to poor diet diversity and low-quality dietary protein. AA deficiencies in early life can cause reductions in linear growth and cognition. Ensuring AA adequacy in diets, particularly through inclusion of nutrient-dense animal source foods from 6 to 23 months, is strongly encouraged in LMICs in order to compensate for less than optimal growth during complementary feeding.


Reinforcement of immunity in Saimiri monkeys following immunization with irradiated sporozoites of Plasmodium vivax.
To determine the duration of immunity to Plasmodium vivax following immunization, six Saimiri sciureus boliviensis monkeys were vaccinated with irradiated sporozoites of P. vivax and challenged multiple times with sporozoites. Over a period of almost four years, complete protection from repeated challenge with infective sporozoites was demonstrated in one monkey; protection in two monkeys was obtained on eight of nine occasions, in one monkey on seven of nine occasions, in one monkey on six or nine occasions, and in one monkey on four of eight occasions. Five of six monkeys were protected against infection during the last six challenges. Inoculation with blood-stage parasites at the end of the trial indicated that all animals were susceptible to infection. These results suggest that protection against sporozoite challenge may be strongly reinforced by subsequent exposure to viable sporozoites.


NMR chemical shift powder pattern recoupling at high spinning speed and theoretical tensor evaluation applied to silk fibroin.
The NMR pulse sequence RAI (recoupling of anisotropy information) has been improved to obtain powder patterns at high MAS spinning speeds. The 2D iso-aniso experiment displays the static chemical shift spectra on the indirect dimension and the MAS spectra on the direct dimension; hence overlapping chemical shift tensor patterns can be well resolved. This efficient technique is applicable to compounds containing (13)C sp(3) (C(alpha), C(beta)) and sp(2) (C=O) sites with higher chemical shift (CS) anisotropy (CSA), and the reliability of the method was tested here on the (13)C chemical shift tensors of polycrystalline glycine, alanine, and serine. Subsequently, the same experiment was applied to the native silk protein fibroin from Bombyx mori, which consists mainly of these three amino acids. Molecular dynamics (MD) simulations of the silk II crystal structure of Takahashi et al. (Takahashi et al. Int. J. Biol. Macromol. 1999, 24, 127-138) were carried out to study the influence of motions on the chemical shift tensors. The (13)C chemical shift tensors were calculated using the bond polarization theory BPT on 200 structures created by an MD simulation. Very good agreement of the theoretical chemical shift anisotropy values with the experimental NMR results was obtained. The tensor orientations in the protein structure could thus be reliably derived.


Favorable effects of vildagliptin on metabolic and cognitive dysfunctions in streptozotocin-induced diabetic rats.
Progression of diabetes mellitus is accompanied by metabolic disorders together with psychological deficits including cognitive dysfunctions. Herein, we used a murine streptozotocin (STZ)-induced diabetes to investigate the beneficial effects of vildagliptin not only on metabolic abnormalities, but also on diabetes-induced cognitive decline. Sixty rats were divided randomly and equally into 2 groups; one remains normal and the other serves as STZ- induced diabetic. Both groups were further divided equally into 2 groups; one received vehicle and the other received oral vildagliptin for 8 weeks. Cognitive behavior was assessed using novel object recognition test. Blood samples were collected to measure metabolic parameters and dipeptidyl peptidase (DPP)-IV activity. Brains were removed and investigated for the levels of inflammatory and oxidative stress markers malondialdehyde (MDA), superoxide dismutase (SOD) and tumor necrosis factor-α (TNF-α), in addition to brain-derived neurotrophic factor (BDNF) and relative expression of nuclear factor kappa B (NF-κB)/p65. Treatment of STZ-induced diabetic rats with vildagliptin increased their body weight and corrected diabetes-induced memory and learning impairment. Moreover, vildagliptin significantly decreased serum levels of glucose and lipids (except high density lipoprotein) together with brain MDA, TNF-α, serum DPP-IV activities and NF-κB/p65 gene expression. On the other hand, vildagliptin significantly increased brain BDNF, SOD as well as serum insulin. Results suggested that vildagliptin has a protective role in counteracting both metabolic abnormalities and memory deficits in diabetic rats, possibly via its anti-hyperglycemic, anti-inflammatory, antioxidant effects, together with reduction of brain NF-κB/p65 over expression.


The impact of a spiritual legacy intervention in patients with brain cancers and other neurologic illnesses and their support persons.
The objectives were to assess the feasibility of using a novel, comprehensive chaplain-led spiritual life review interview to develop a personal Spiritual Legacy Document (SLD) for persons with brain tumors and other neurodegenerative diseases and to describe spiritual well-being (SWB), spiritual coping, and quality of life (QOL) of patients and their support persons (SP) before and after receipt of the SLD. Patient-SP pairs were enrolled over a 2-year period. Assessments included the Functional Assessment of Chronic Illness Therapy-Spiritual Expanded Version, Brief Religious Coping Scale, Brief COPE Inventory, and QOL Linear Analog Scale. Baseline assessments were completed prior to an audio-recorded spiritual life review interview with a chaplain. Thirty-two patient/SP pairs were enrolled; 27 completed baseline assessments and the interview. Twenty-four reviewed their SLD and were eligible for follow-up. A total of 15 patients and 12 SPs completed the 1-month follow-up; 10 patients and seven SPs completed the 3-month follow-up. Patients endorsed high levels of SWB and spiritual coping at baseline. Both patients and SPs evidenced improvement on several aspects of SWB, spiritual coping, and QOL at 1 month, but patients' decreased financial well-being was also observed. Patients and SPs demonstrated favorable changes in peacefulness and positive religious coping at both time points. A chaplain-led spiritual life review is a feasible intervention for patients with neurodegenerative disease and results in beneficial effects on patients and SPs. Copyright © 2015 John Wiley & Sons, Ltd.


Proposal of new body composition prediction equations from bioelectrical impedance for Indonesian men.
Bioelectrical impedance analysis (BIA) is one of the most affordable and feasible body composition assessment techniques for clinical and field settings. However, it is important to use an equation appropriate for the study population. This study aimed to propose and validate prediction equations to estimate body composition using BIA for Indonesian men. Total body water (TBW), fat-free mass (FFM) and fat mass (FM) were determined using the deuterium dilution technique in 292 Indonesian males. Participants were divided equally into development and validation groups to develop prediction equations and to cross-validate the proposed prediction equations, respectively. In addition, selected prediction equations using BIA were cross-validated. The proposed BIA equations were valid in our cross-validation samples. The best performance equations obtained from the absolute measure of body composition (TBW, FFM and FM) showed that r ranged between 0.89 and 0.91 and standard error of the estimate=1.8-2.6 kg. Cross-validation analysis indicated that the proposed equations had a bias of 0.1-0.3 kg, pure error of 1.3-1.8 kg and limits of agreement (mean difference±1.96 s.d.) of -0.26 to 0.13±4.09 to 5.59 kg. Among existing prediction equations examined, those by Deurenberg et al. (1989) and Lukaski et al. (1987) significantly overestimated FM by 4.0 and 3.2 kg, respectively, whereas the equation by Deurenberg et al. (1991) significantly (P<0.001) underestimated FFM by 5.0 kg compared with the reference FFM. The new BIA prediction equations may provide more precise and accurate estimation of body composition in Indonesian men than the existing equations.


TiO2 nanoparticles suppress Escherichia coli cell division in the absence of UV irradiation in acidic conditions.
TiO(2) nanoparticles (NPs) activated by UV irradiation are known to have a bactericidal effect. In this study we report the details of TiO(2) NPs influence on the colony-forming capacity of E. coli in the dark at pH 4.0-4.5. At this pH the bacterial cells are negatively charged and TiO(2) NPs present a positive charge. A 60 min contact between E. coli with TiO(2) at concentrations of 0.02-0.2 mg/mL led to a reduction of E. coli cell number from 10(8) to 10(4)CFU/mL. After the reduction the system remains unchanged during the subsequent incubation. The observed reduction was a function on the initial E. coli concentration. In the presence of 0.04 mg/mL TiO(2) the colony-forming units (CFU) reduction after 60 min was of four-five orders of magnitude when the initial concentration was 10(8) cells/mL. But when starting with an E. coli concentration of 10(7) cells/mL the cell number reduction was less than one order of magnitude. Less than one order of magnitude cell number reduction was also observed for suspensions of E. coli 10(8) cells/mL and 0.002 mg/mL of TiO(2). The bacteria number reduction was always accompanied by the formation of cell aggregates. During cell incubation with TiO(2), the pH of the suspension increased, but did not reach the TiO(2) isoelectric point (IEP). E. coli cells stained with the fluorescent dye acridine orange (AO) showed that the fluorescence of single cells remained unchanged after incubation in the presence of TiO(2). The color change of fluorescence was revealed only in aggregated cells. This suggests changes in the physiologic state of E. coli incorporated into the aggregates. Aggregates of E. coli occur due to the electrostatic interaction between TiO(2) NPs and the bacterial cell surface. A hypothesis is suggested in this study to explain the CFU reduction and the retention of a certain irreducible number of cells capable of further division in the suspension in the presence of TiO(2) in the dark.


Long noncoding intronic RNAs are differentially expressed in primary and metastatic pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) is known by its aggressiveness and lack of effective therapeutic options. Thus, improvement in current knowledge of molecular changes associated with pancreatic cancer is urgently needed to explore novel venues of diagnostics and treatment of this dismal disease. While there is mounting evidence that long noncoding RNAs (lncRNAs) transcribed from intronic and intergenic regions of the human genome may play different roles in the regulation of gene expression in normal and cancer cells, their expression pattern and biological relevance in pancreatic cancer is currently unknown. In the present work we investigated the relative abundance of a collection of lncRNAs in patients' pancreatic tissue samples aiming at identifying gene expression profiles correlated to pancreatic cancer and metastasis. Custom 3,355-element spotted cDNA microarray interrogating protein-coding genes and putative lncRNA were used to obtain expression profiles from 38 clinical samples of tumor and non-tumor pancreatic tissues. Bioinformatics analyses were performed to characterize structure and conservation of lncRNAs expressed in pancreatic tissues, as well as to identify expression signatures correlated to tissue histology. Strand-specific reverse transcription followed by PCR and qRT-PCR were employed to determine strandedness of lncRNAs and to validate microarray results, respectively. We show that subsets of intronic/intergenic lncRNAs are expressed across tumor and non-tumor pancreatic tissue samples. Enrichment of promoter-associated chromatin marks and over-representation of conserved DNA elements and stable secondary structure predictions suggest that these transcripts are generated from independent transcriptional units and that at least a fraction is under evolutionary selection, and thus potentially functional.Statistically significant expression signatures comprising protein-coding mRNAs and lncRNAs that correlate to PDAC or to pancreatic cancer metastasis were identified. Interestingly, loci harboring intronic lncRNAs differentially expressed in PDAC metastases were enriched in genes associated to the MAPK pathway. Orientation-specific RT-PCR documented that intronic transcripts are expressed in sense, antisense or both orientations relative to protein-coding mRNAs. Differential expression of a subset of intronic lncRNAs (PPP3CB, MAP3K14 and DAPK1 loci) in metastatic samples was confirmed by Real-Time PCR. Our findings reveal sets of intronic lncRNAs expressed in pancreatic tissues whose abundance is correlated to PDAC or metastasis, thus pointing to the potential relevance of this class of transcripts in biological processes related to malignant transformation and metastasis in pancreatic cancer.


Providing underserved patients with medical homes: assessing the readiness of safety-net health centers.
Enthusiasm for the patient-centered medical home model is growing, yet initial research is scant, showing that true transformation is challenging and that meaningful improvements in care delivery, efficiency, and health outcomes take time and sustained investment. This brief surveys safety-net health centers to determine their potential to become medical homes. Safety-net health centers that provide vulnerable and low-income populations with comprehensive primary care have unique opportunities for successful transformation, but also face challenges. For example, nearly half of the health centers surveyed do not have a process for scheduling patients with a personal provider or have an existing process that needs improvement; two-thirds do not have a process for same-day scheduling or have a process that needs improvement. Survey data also show that health centers that employed team-based care were more likely to have instituted patient access and communications processes, relative to those without team-based care.


Inclusion of Almonds in a Cholesterol-Lowering Diet Improves Plasma HDL Subspecies and Cholesterol Efflux to Serum in Normal-Weight Individuals with Elevated LDL Cholesterol.
Background: Almonds may increase circulating HDL cholesterol when substituted for a high-carbohydrate snack in an isocaloric diet, yet little is known about the effects on HDL biology and function.Objective: The objective was to determine whether incorporating 43 g almonds/d in a cholesterol-lowering diet would improve HDL subspecies and function, which were secondary study outcomes.Methods: In a randomized, 2-period, crossover, controlled-feeding study, a diet with 43 g almonds/d (percentage of total energy: 51% carbohydrate, 16% protein, and 32% total and 8% saturated fat) was compared with a similar diet with an isocaloric muffin substitution (58% carbohydrate, 15% protein, and 26% total and 8% saturated fat) in men and women with elevated LDL cholesterol. Plasma HDL subspecies and cholesterol efflux from J774 macrophages to human serum were measured at baseline and after each diet period. Diet effects were examined in all participants (n = 48) and in normal-weight (body mass index: <25; n = 14) and overweight or obese (≥25; n = 34) participants by using linear mixed models.Results: The almond diet, compared with the control diet, increased α-1 HDL [mean ± SEM: 26.7 ± 1.5 compared with 24.3 ± 1.3 mg apolipoprotein A-I (apoA-I)/dL; P = 0.001]. In normal-weight participants, the almond diet, relative to the control diet, increased α-1 HDL (33.7 ± 3.2 compared with 28.4 ± 2.6 mg apoA-I/dL), the α-1 to pre-β-1 ratio [geometric mean (95% CI): 4.3 (3.3, 5.7) compared with 3.1 (2.4, 4.0)], and non-ATP-binding cassette transporter A1 cholesterol efflux (8.3% ± 0.4% compared with 7.8% ± 0.3%) and decreased pre-β-2 (3.8 ± 0.4 compared with 4.6 ± 0.4 mg apoA-I/dL) and α-3 (23.5 ± 0.9 compared with 26.9 ± 1.1 mg apoA-I/dL) HDL (P < 0.05). No diet effects were observed in the overweight or obese group.Conclusions: Substituting almonds for a carbohydrate-rich snack within a lower-saturated-fat diet may be a simple strategy to maintain a favorable circulating HDL subpopulation distribution and improve cholesterol efflux in normal-weight individuals with elevated LDL cholesterol. This trial was registered at clinicaltrials.gov as NCT01101230.


Thermotoga maritima NusG: domain interaction mediates autoinhibition and thermostability.
NusG, the only universally conserved transcription factor, comprises an N- and a C-terminal domain (NTD, CTD) that are flexibly connected and move independently in Escherichia coli and other organisms. In NusG from the hyperthermophilic bacterium Thermotoga maritima (tmNusG), however, NTD and CTD interact tightly. This closed state stabilizes the CTD, but masks the binding sites for the interaction partners Rho, NusE and RNA polymerase (RNAP), suggesting that tmNusG is autoinhibited. Furthermore, tmNusG and some other bacterial NusGs have an additional domain, DII, of unknown function. Here we demonstrate that tmNusG is indeed autoinhibited and that binding to RNAP may stabilize the open conformation. We identified two interdomain salt bridges as well as Phe336 as major determinants of the domain interaction. By successive weakening of this interaction we show that after domain dissociation tmNusG-CTD can bind to Rho and NusE, similar to the Escherichia coli NusG-CTD, indicating that these interactions are conserved in bacteria. Furthermore, we show that tmNusG-DII interacts with RNAP as well as nucleic acids with a clear preference for double stranded DNA. We suggest that tmNusG-DII supports tmNusG recruitment to the transcription elongation complex and stabilizes the tmNusG:RNAP complex, a necessary adaptation to high temperatures.


Potent combinations of beta-lactam antibiotics using the beta-lactamase inhibition principle.
Several penicillins known to be stable to enterobacterial beta-lactamases were tested in combination with beta-lactamase-sensitive penicillins and cephalosporins in a turbidimetric system. Nafcillin was found to be the best beta-lactamase inhibitor amongst agents presently available commercially, but the related, new semi-synthetic penicillin BRL 1437 (2-isopropoxy-1-naphthylpenicillin) was consistently found to be superior. Using 103 ampicillin-resistant coliform bacilli and antibiotic levels achievable in urine, cephalothin or cephaloridine alone achieved long-term suppression of growth (greater than 20 h) of 16 and 13% of strains, respectively, while the additional presence of BRL 1437 suppressed growth for longer than 20 h of 81% of the remaining strains. Even where 'success' was not achieved according to these stringent criteria, regrowth was significantly delayed by the presence of BRL 1437. Suppression of growth for longer than 20 h by BRL 1437 plus cephalothin was achieved with all of the 46 Escherichia coli strains tested. Antibiotic combinations were also studied in an in vitro model which stimulates the hydrokinetic features of the urinary bladder. Suppression of the growth of two highly resistant E. coli strains was achieved in this system, for therapeutically acceptable periods of time, with combinations of cephalothin or cephaloridine with BRL 1437, but not nafcillin.


Population genetics of immune-related multilocus copy number variation in Native Americans.
While multiallelic copy number variation (mCNV) loci are a major component of genomic variation, quantifying the individual copy number of a locus and defining genotypes is challenging. Few methods exist to study how mCNV genetic diversity is apportioned within and between populations (i.e. to define the population genetic structure of mCNV). These inferences are critical in populations with a small effective size, such as Amerindians, that may not fit the Hardy-Weinberg model due to inbreeding, assortative mating, population subdivision, natural selection or a combination of these evolutionary factors. We propose a likelihood-based method that simultaneously infers mCNV allele frequencies and the population structure parameter f, which quantifies the departure of homozygosity from the Hardy-Weinberg expectation. This method is implemented in the freely available software CNVice, which also infers individual genotypes using information from both the population and from trios, if available. We studied the population genetics of five immune-related mCNV loci associated with complex diseases (beta-defensins, CCL3L1/CCL4L1, FCGR3A, FCGR3B and FCGR2C) in 12 traditional Native American populations and found that the population structure parameters inferred for these mCNVs are comparable to but lower than those for single nucleotide polymorphisms studied in the same populations.


Mechanism of ion transport in perfluoropolyether electrolytes with a lithium salt.
Perfluoropolyethers (PFPEs) are polymer electrolytes with fluorinated carbon backbones that have high flash points and have been shown to exhibit moderate conductivities and high cation transference numbers when mixed with lithium salts. Ion transport in four PFPE electrolytes with different endgroups was characterized by differential scanning calorimetry (DSC), ac impedance, and pulsed-field gradient NMR (PFG-NMR) as a function of salt concentration and temperature. In spite of the chemical similarity of the electrolytes, salt diffusion coefficients measured by PFG-NMR and the glass transition temperature measured by DSC appear to be uncorrelated to ionic conductivity measured by ac impedance. We calculate a non-dimensional parameter, β, that depends on the salt diffusion coefficients and ionic conductivity. We also use the Vogel-Tammann-Fulcher relationship to fit the temperature dependence of conductivity. We present a linear relationship between the prefactor in the VTF fit and β; both parameters vary by four orders of magnitude in our experimental window. Our analysis suggests that transport in electrolytes with low dielectric constants (low β) is dictated by ion hopping between clusters.


Potential synergy of phytochemicals in cancer prevention: mechanism of action.
Epidemiological studies have consistently shown that regular consumption of fruits and vegetables is strongly associated with reduced risk of developing chronic diseases, such as cancer and cardiovascular disease. It is now widely believed that the actions of the antioxidant nutrients alone do not explain the observed health benefits of diets rich in fruits and vegetables, because taken alone, the individual antioxidants studied in clinical trials do not appear to have consistent preventive effects. Work performed by our group and others has shown that fruits and vegetable phytochemical extracts exhibit strong antioxidant and antiproliferative activities and that the major part of total antioxidant activity is from the combination of phytochemicals. We proposed that the additive and synergistic effects of phytochemicals in fruits and vegetables are responsible for these potent antioxidant and anticancer activities and that the benefit of a diet rich in fruits and vegetables is attributed to the complex mixture of phytochemicals present in whole foods. This explains why no single antioxidant can replace the combination of natural phytochemicals in fruits and vegetables to achieve the health benefits. The evidence suggests that antioxidants or bioactive compounds are best acquired through whole-food consumption, not from expensive dietary supplements. We believe that a recommendation that consumers eat 5 to 10 servings of a wide variety of fruits and vegetables daily is an appropriate strategy for significantly reducing the risk of chronic diseases and to meet their nutrient requirements for optimum health.


The seasonality of phosphorus transfers from land to water: implications for trophic impacts and policy evaluation.
The Nitrates Directive regulations are a Programme of Measures under the EU Water Framework Directive to protect waters from agricultural transfers of nitrogen and phosphorus. Soil phosphorus management to an agronomic optimum and closed winter periods for organic and inorganic fertiliser amendments are among a suite of policy measures to curtail diffuse pollution at catchment scale. In this investigation, two intensive grassland and two arable catchments (7-12 km(2)) in the Republic of Ireland were studied to link a high resolution spatial survey (≤2 ha) of soil P availability with P delivery in receiving rivers; monitored on a sub-hourly basis over one year. Data indicated that source risk, as defined by soil P availability and organic P loading, was less important than mobilisation and hydrological transfer potential which increased delivery due to runoff flashiness as described by a hydrological metric during the winter. Overall, however, annual TP loads were low to moderate (0.175 to 0.785 kg ha(-1) yr(-1)). The data also highlighted, without exception, the influences of summer background P loading and subsequent ecologically significant P concentrations from persistent point sources. This may have implications for expected ecological status and recovery in these catchments, which appeared more at risk in catchments with little buffering in terms of summer base flow dilution. Wetter winters and drier summers under climate change scenarios would likely increase stream P concentrations both during storms and during baseflows and would be particularly magnified in those catchments with flashy runoff and suppressed baseflow. These seasonal insights into source-to-delivery functions and risk (re)assessment were only possible with high resolution (spatial and temporal) data collection and will be important in influencing expectations of policies that are evaluated at larger scales but with coarser resolution sampling.


[The risk of the military profession in the city of Rio de Janeiro in 'peace time': the perception of the troop].
This study aims at determining the risk perception of Army Military concerning common activities at Military Units of troop in Rio de Janeiro. This mapping might allow taking preventive action when preparing these professionals, reducing their vulnerability. The present study is descriptive and used a qualitative methodological approach. The informers were selected into four military units and divided in three groups: eight officers, eight sergeants and eight corporal/soldiers, summing up 24 informers. It was possible to identify three risk categories noticed by the military in their common Military Units activities: (a) The risk of being a military in a city where urban violence reached the level there is in Rio de Janeiro city; (b) the risk of accidents and; (c) the risk of injuries or chronic pathology related to the exposure to activities that need proper protection, noticed by a number of informants. It can be concluded that the local violence has a great influence on the health of the military, due to the fact that modifies substantially the daily routines and their emotions during the work and that the perception the military have of the risks of their activities is focused on the present moment.


Reduction in saturated fat intake for cardiovascular disease.
Reducing saturated fat reduces serum cholesterol, but effects on other intermediate outcomes may be less clear. Additionally, it is unclear whether the energy from saturated fats eliminated from the diet are more helpfully replaced by polyunsaturated fats, monounsaturated fats, carbohydrate or protein. To assess the effect of reducing saturated fat intake and replacing it with carbohydrate (CHO), polyunsaturated (PUFA), monounsaturated fat (MUFA) and/or protein on mortality and cardiovascular morbidity, using all available randomised clinical trials. We updated our searches of the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE (Ovid) and Embase (Ovid) on 15 October 2019, and searched Clinicaltrials.gov and WHO International Clinical Trials Registry Platform (ICTRP) on 17 October 2019. Included trials fulfilled the following criteria: 1) randomised; 2) intention to reduce saturated fat intake OR intention to alter dietary fats and achieving a reduction in saturated fat; 3) compared with higher saturated fat intake or usual diet; 4) not multifactorial; 5) in adult humans with or without cardiovascular disease (but not acutely ill, pregnant or breastfeeding); 6) intervention duration at least 24 months; 7) mortality or cardiovascular morbidity data available. Two review authors independently assessed inclusion, extracted study data and assessed risk of bias. We performed random-effects meta-analyses, meta-regression, subgrouping, sensitivity analyses, funnel plots and GRADE assessment. We included 15 randomised controlled trials (RCTs) (16 comparisons, ~59,000 participants), that used a variety of interventions from providing all food to advice on reducing saturated fat. The included long-term trials suggested that reducing dietary saturated fat reduced the risk of combined cardiovascular events by 21% (risk ratio (RR) 0.79; 95% confidence interval (CI) 0.66 to 0.93, 11 trials, 53,300 participants of whom 8% had a cardiovascular event, I² = 65%, GRADE moderate-quality evidence). Meta-regression suggested that greater reductions in saturated fat (reflected in greater reductions in serum cholesterol) resulted in greater reductions in risk of CVD events, explaining most heterogeneity between trials. The number needed to treat for an additional beneficial outcome (NNTB) was 56 in primary prevention trials, so 56 people need to reduce their saturated fat intake for ~four years for one person to avoid experiencing a CVD event. In secondary prevention trials, the NNTB was 32. Subgrouping did not suggest significant differences between replacement of saturated fat calories with polyunsaturated fat or carbohydrate, and data on replacement with monounsaturated fat and protein was very limited. We found little or no effect of reducing saturated fat on all-cause mortality (RR 0.96; 95% CI 0.90 to 1.03; 11 trials, 55,858 participants) or cardiovascular mortality (RR 0.95; 95% CI 0.80 to 1.12, 10 trials, 53,421 participants), both with GRADE moderate-quality evidence. There was little or no effect of reducing saturated fats on non-fatal myocardial infarction (RR 0.97, 95% CI 0.87 to 1.07) or CHD mortality (RR 0.97, 95% CI 0.82 to 1.16, both low-quality evidence), but effects on total (fatal or non-fatal) myocardial infarction, stroke and CHD events (fatal or non-fatal) were all unclear as the evidence was of very low quality. There was little or no effect on cancer mortality, cancer diagnoses, diabetes diagnosis, HDL cholesterol, serum triglycerides or blood pressure, and small reductions in weight, serum total cholesterol, LDL cholesterol and BMI. There was no evidence of harmful effects of reducing saturated fat intakes. The findings of this updated review suggest that reducing saturated fat intake for at least two years causes a potentially important reduction in combined cardiovascular events. Replacing the energy from saturated fat with polyunsaturated fat or carbohydrate appear to be useful strategies, while effects of replacement with monounsaturated fat are unclear. The reduction in combined cardiovascular events resulting from reducing saturated fat did not alter by study duration, sex or baseline level of cardiovascular risk, but greater reduction in saturated fat caused greater reductions in cardiovascular events.


Metabolic activation of 2,6-dichlorobenzonitrile, an olfactory-specific toxicant, by rat, rabbit, and human cytochromes P450.
The herbicide 2,6-dichlorobenzonitrile (DCBN) is known to cause tissue-specific toxicity at very low doses in the olfactory mucosa of rodents. The toxicity of DCBN is reportedly cytochrome P450 (P450) dependent, but the isoforms involved have not been identified, and the effects of this agent on humans are not known. In the present study, DCBN metabolism was examined with microsomes and with purified P450s in a reconstituted system. Rat and rabbit olfactory microsomes act on DCBN to form DCBN-protein adducts as well as two metabolite peaks, designated M1 and M2, identified through high performance liquid chromatography with radiometric detection. The activity of rat olfactory microsomes in DCBN metabolism is much higher than that of liver or lung microsomes. Of seven purified rabbit P450s known to be expressed in the olfactory mucosa, including 1A2, 2A10/11, 2B4, 2E1, 2G1, and 3A6, the 2A10/11 preparation is the most active, producing M2 as well as DCBN-protein adducts; P450 2E1 is the only other active isoform. The addition of purified epoxide hydrolase (EC 220.127.116.11) to the reconstituted enzyme system leads to the formation of M1 and decreased formation of M2. It seems that M1 and M2 are derived from an epoxide intermediate that also forms covalent protein adducts. Gas chromatography- and liquid chromatography-mass spectrometry analyses of nasal microsomal DCBN metabolites and DCBN-glutathione conjugates indicated that the major reactive intermediate may be 2,3-oxo-DCBN and that M1 may be 2,3-dihydroxy-6-chlorobenzonitrile, whereas M2 may correspond to a monohydroxy-DCBN. Interestingly, heterologously expressed human P450s 2A6 and 2E1, but not 1A2, are active in the metabolism of DCBN, forming protein adducts as well as M2. Thus, the preferential expression of P450s of the 2A subfamily in olfactory tissue suggests a molecular basis for the tissue-specific toxicity of the herbicide and may have important implications for risk assessment in humans.


Loss of cancer drug activity in colon cancer HCT-116 cells during spheroid formation in a new 3-D spheroid cell culture system.
Clinically relevant in vitro methods are needed to identify new cancer drugs for solid tumors. We report on a new 3-D spheroid cell culture system aimed to mimic the properties of solid tumors in vivo. The colon cancer cell lines HCT-116 wt and HCT-116 wt/GFP were grown as monolayers and for 3 or 6 days on 96-well NanoCulture® plates to form spheroids. Expression of surface markers, genes and hypoxia were assessed to characterize the spheroids and drug induced cytotoxicity was evaluated based on fluorescein diacetate (FDA) conversion by viable cells to fluorescent fluorescein or by direct measurement of fluorescence of GFP marked cells after a 72 h drug incubation. The cells reproducibly formed spheroids in the NanoCulture® plates with tight cell-attachment after 6 days. Cells in spheroids showed geno- and phenotypical properties reminiscent of hypoxic stem cells. Monolayer cultured cells were sensitive to standard and investigational drugs, whereas the spheroids gradually turned resistant. Similar results for cytotoxicity were observed using simplified direct measurement of fluorescence of GFP marked cells compared with FDA incubation. In conclusion, this new 3-D spheroid cell culture system provides a convenient and clinically relevant model for the identification and characterization of cancer drugs for solid tumors.


Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly.
Nitric oxide contrasts with most intercellular messengers because it diffuses rapidly and isotropically through most tissues with little reaction but cannot be transported through the vasculature due to rapid destruction by oxyhemoglobin. The rapid diffusion of nitric oxide between cells allows it to locally integrate the responses of blood vessels to turbulence, modulate synaptic plasticity in neurons, and control the oscillatory behavior of neuronal networks. Nitric oxide is not necessarily short lived and is intrinsically no more reactive than oxygen. The reactivity of nitric oxide per se has been greatly overestimated in vitro because no drain is provided to remove nitric oxide. Nitric oxide persists in solution for several minutes in micromolar concentrations before it reacts with oxygen to form much stronger oxidants like nitrogen dioxide. Nitric oxide is removed within seconds in vivo by diffusion over 100 microns through tissues to enter red blood cells and react with oxyhemoglobin. The direct toxicity of nitric oxide is modest but is greatly enhanced by reacting with superoxide to form peroxynitrite (ONOO-). Nitric oxide is the only biological molecule produced in high enough concentrations to out-compete superoxide dismutase for superoxide. Peroxynitrite reacts relatively slowly with most biological molecules, making peroxynitrite a selective oxidant. Peroxynitrite modifies tyrosine in proteins to create nitrotyrosines, leaving a footprint detectable in vivo. Nitration of structural proteins, including neurofilaments and actin, can disrupt filament assembly with major pathological consequences. Antibodies to nitrotyrosine have revealed nitration in human atherosclerosis, myocardial ischemia, septic and distressed lung, inflammatory bowel disease, and amyotrophic lateral sclerosis.


Aeromicrobium halocynthiae sp. nov., a taurocholic acid-producing bacterium isolated from the marine ascidian Halocynthia roretzi.
A marine bacterium, strain KME 001(T), was isolated from the siphon tissue of a marine ascidian, Halocynthia roretzi, collected off the coast of Gangneung, Korea. Strain KME 001(T) was a Gram-positive, aerobic, non-spore-forming, rod-shaped and non-motile bacterium. Phylogenetic analysis using 16S rRNA gene sequences revealed that strain KME 001(T) clustered with the genus Aeromicrobium and was closely related to Aeromicrobium ginsengisoli, Aeromicrobium erythreum and Aeromicrobium ponti with 97.7, 97.6 and 97.5 % sequence similarities, respectively. The strain was capable of growth at a variety of temperatures (10-42°C) and over a broad pH range (5.0-10.0). NaCl was required for robust growth of the strain. The diagnostic diamino acid of the cell-wall peptidoglycan was ll-diaminopimelic acid. The predominant menaquinone was MK-9(H(4)). The predominant fatty acids were C(18 : 1)ω9c, C(16 : 0) and 10-methyl C(18 : 0). The DNA-DNA hybridization analyses showed that DNA-DNA relatedness values between strain KME 001(T) and its nearest neighbours, A. ginsengisoli KCTC 19207(T), A. erythreum KCCM 41104(T) and A. ponti KACC 20565(T), were 49.6, 57.1 and 63.5 %, respectively. The DNA G+C content of strain KME 001(T) was 75.9mol%. Chemical investigation of the liquid culture medium of strain KME 001(T) led to the isolation of taurocholic acid as a major secondary metabolite. On the basis of phylogenetic and phenotypic data, strain KME 001(T) is classified as representing a novel species of the genus Aeromicrobium, for which the name Aeromicrobium halocynthiae sp. nov. is proposed. The type strain is KME 001(T) (=JCM 15749(T)=KCCM 90079(T)).


Design of hemi-urchin shaped ZnO nanostructures for broadband and wide-angle antireflection coatings.
We study theoretically and experimentally the hemi-urchin shaped zinc oxide (ZnO) nanostructures for broadband and wide-angle antireflection coatings. The antireflective characteristics of hemi-urchin shaped ZnO nanostructures, which can be formed by integrating one-dimensional (1D) nanostructures (i.e., nanorods) on the periodic 2D structural architecture, are investigated. The optimization process is performed using a rigorous coupled-wave analysis method in terms of the order of taper of Si subwavelength gratings (SWGs) as a 2D structural architecture, the geometry of Si SWGs, and the height/size of ZnO nanorods. To simply test an experimental feasibility, a hemi-urchin shaped ZnO nanostructure is fabricated by hydrothermally growing ZnO nanorods on the periodic Si SWG structure. The angle-dependent reflectance of the hemi-urchin shaped ZnO nanostructures on the Si SWG is compared with that of the vertically aligned ZnO nanorod arrays on the Si substrate. The optimized hemi-urchin shaped ZnO nanostructure can significantly improve the antireflective property by suppressing the surface reflection over a broad spectrum and a wide range of angles of light incidence, indicating a reasonable agreement with the experimental results.


Transcranial Magnetic Stimulation Over the Human Medial Posterior Parietal Cortex Disrupts Depth Encoding During Reach Planning.
Accumulating evidence supports the view that the medial part of the posterior parietal cortex (mPPC) is involved in the planning of reaching, but while plenty of studies investigated reaching performed toward different directions, only a few studied different depths. Here, we investigated the causal role of mPPC (putatively, human area V6A-hV6A) in encoding depth and direction of reaching. Specifically, we applied single-pulse transcranial magnetic stimulation (TMS) over the left hV6A at different time points while 15 participants were planning immediate, visually guided reaching by using different eye-hand configurations. We found that TMS delivered over hV6A 200 ms after the Go signal affected the encoding of the depth of reaching by decreasing the accuracy of movements toward targets located farther with respect to the gazed position, but only when they were also far from the body. The effectiveness of both retinotopic (farther with respect to the gaze) and spatial position (far from the body) is in agreement with the presence in the monkey V6A of neurons employing either retinotopic, spatial, or mixed reference frames during reach plan. This work provides the first causal evidence of the critical role of hV6A in the planning of visually guided reaching movements in depth.


Distribution of eschars on the body of scrub typhus patients: a prospective study.
Eschar is an important finding for the diagnosis of scrub typhus. The IFA test for possible scrub typhus was performed. The presence or absence of eschar was thoroughly examined. Among the 176 scrub typhus cases confirmed by IFA, 162 (92.0%) cases had eschar; 128 patients (79.5%) had eschars on the front of the body. Eschars were primarily detected in males within 30 cm below the umbilicus (19 patients, 35.8%). Distributions on the lower extremities and the front chest above the umbilicus were 22.6% (12 patients) and 20.8% (11 patients), respectively. A different pattern was seen in females. The most prevalent area was the front chest above the umbilicus, which accounted for 40.7% (44 patients) of all the detected eschars. Our study is the first report of a schematic diagram that shows the differences between the males and females with respect to eschar location in scrub typhus patients.


Lung function, respiratory symptoms, and the menopausal transition.
There is limited information on potential changes in respiratory health when women enter the menopausal transition. We sought to investigate whether the menopausal transition is related to lung function and asthma and whether body mass index (BMI) modifies associations. Four thousand two hundred fifty-nine women from 21 centers (ECRHS II, 2002) responded to a questionnaire concerning women's health. Women aged 45 to 56 years not using exogenous sex hormones (n = 1274) were included in the present analysis. Lung function measurements (n = 1120) and serum markers of hormonal status (follicle-stimulating hormone, luteinizing hormone, and estradiol; n = 710) were available. Logistic and linear regression analyses were adjusted for BMI, age, years of education, smoking status, center, and height. Women not menstruating for the last 6 months (n = 432, 34%) had significantly lower FEV(1) values (-120 mL [95% CI, -177 to -63]), lower forced vital capacity values (-115 mL [95% CI, -181 to -50]), and more respiratory symptoms (odds ratio [OR], 1.82 [95% CI, 1.27-2.61]) than those menstruating regularly. Results were similar when restricting analyses to those who never smoked. Associations were significantly stronger in women with BMIs of less than 23 kg/m(2) (respiratory symptoms: OR, 4.07 [95% CI, 1.88-8.80]; FEV(1) adjusted difference: -166 [95% CI, -263 to -70]) than in women with BMIs of 23 to 28 kg/m(2) (respiratory symptoms: OR, 1.10 [95% CI, 0.61-1.97], P(interaction): .04; FEV(1) adjusted difference, -54 [95% CI, -151 to 43], P(interaction) = .06). Menopause is associated with lower lung function and more respiratory symptoms, especially among lean women.


Visual and semiquantitative analyses for F-18 fluorodeoxyglucose PET scanning in pulmonary nodules 1 cm to 3 cm in size.
While visual assessment is the simplest way to evaluate positron emission tomography (PET) with fluorodeoxyglucose (FDG), faintly positive nodules are often difficult to evaluate. We performed visual and semiquantitative analyses of pulmonary nodules from 1 to 3 cm in size to determine the optimal method of analysis for PET data, especially for faintly positive nodules on visual assessment. Positron emission tomography data were analyzed for 161 pulmonary nodules from 1 to 3 cm in size (108 malignant and 53 benign nodules). On visual assessment, FDG uptake by the nodules was classified into three grades in comparison with mediastinal blood pool, (ie, definitely positive, faintly positive, and negative). In addition, FDG uptake was measured by the standard uptake value (SUV), the contrast ratio to the contralateral lung (CR-lung), and the contrast ratio to the cerebellum (CR-brain). Cut-off values for each variable were determined from receiver operating characteristics (ROC) curves, and the values were 2.5 for the SUV, 0.4 for the CR-lung, and 0.25 for the CR-brain. Nodules with FDG uptake above these cut-off values were defined as positive in each method. Visual assessment showed definitely positive for 80 nodules (64 malignant and 16 benign), faintly positive for 22 (17 malignant and 5 benign), and negative for 59 nodules (27 malignant and 32 benign). In the 139 nodules that were definitely positive or negative by visual assessment, there were no significant differences of sensitivity and specificity among the four methods (visual assessment, SUV, CR-lung, and CR-brain). In the 17 malignant nodules that were faintly positive, the SUV did not detect any true-positive nodules, but the CR-lung and CR-brain showed 9 and 5 true-positive nodules, respectively, resulting in a significantly higher sensitivity than the SUV (p < 0.001 and p = 0.02, respectively). Fifteen of 17 faintly positive malignant nodules (88%) were histologically well- or moderately-differentiated adenocarcinoma. The semiquantitative methods (SUV, CR-lung, and CR-brain) do not improve the efficacy of visual assessment alone for nodules graded as definitely positive or negative. However, faintly positive nodules on visual assessment should be evaluated by the CR-lung or CR-brain rather than the SUV, and are often differentiated adenocarcinomas.


MO-D-BRB-03: Comparison of Proton Therapy Institutional Data Collected by the RPC.
To detail and compare data collected during RPC onsite dosimetry review visits at proton therapy centers. The RPC has established a complete review process for proton therapy institutions wishing to participate in NCI-funded clinical trials that includes an on-site dosimetry review visit performed by the RPC. During the visit, the RPC takes measurements that include CT# vs. relative stopping power (RSP) conversion, beam output, depth dose, lateral profiles, QA procedure reviews and anthropomorphic phantom irradiations. The RPC reviewed beam output, depth dose and lateral profiles for 5 specific anatomic treatment sites, reference, prostate, lung, brain, and spine as compared to the institution's measured or treatment planning-derived values. In addition, the RPC has compared results from each institution's proton prostate phantom irradiation. All of the institutions visited had RPC/Institution output ratios that ranged between 0.95 - 1.02 where the acceptance criterion was ±5%. For the CT# to RSP comparison, there was a larger variability. Only two institutions agreed within five percent of the recommended values, while the other five institutions had disagreements of up to 20 percent in the high density (high CT number) region of the conversion curve that may have a clinical impact on dose delivery. For the prostate phantom irradiation, 3 institutions failed to meet the RPC's ±7%/4mm acceptance criteria on the initial attempt, but in the end all 7 sites met the criteria. The proton beam output for 7 proton centers, as measured by the RPC, is comparable (±5%), however, there are large discrepancies in the CT# vs RSP conversion curves used from institution to institution. As a result of the RPC onsite dosimetry review visits, several institutions have modified their procedures and dosimetry parameters to improve proton therapy delivery for NCI funded clinical trials. Work supported by grants CA10953, CA059267, and CA81647 (NCI, DHHS).


Individual kidney blood flow measured with contrast-enhanced first-pass perfusion MR imaging.
The study design was HIPAA-compliant and approved by the Institutional Review Board, with all participants providing signed informed consent prior to the study. The purpose of this study was to prospectively evaluate the feasibility of determining renal blood flow (RBF) by using a technique based on intravenous administration of gadolinium chelate and evaluation of first-pass gadolinium chelate perfusion by using highly accelerated three-dimensional (3D) gradient-echo magnetic resonance (MR) imaging of the kidney in freely breathing subjects. Flow is determined with Kety-Schmidt formalism by modeling the uptake of gadolinium chelate in the kidney prior to its leaving through the venous system. Validation of the gadolinium chelate perfusion technique is based on comparison of values determined for participants with phase-contrast gradient-echo imaging. The model fit to the measured data is excellent over the first 7-8 seconds of gadolinium chelate uptake and diverges after its appearance in the renal vein. The perfusion data analysis technique showed less than 10% interobserver variation. The average difference between phase-contrast and gadolinium chelate perfusion measurements was 0.08 mL/sec (95% confidence interval: -3.73, 3.58) for left and right kidneys. This study demonstrates feasibility of the gadolinium chelate perfusion method for RBF measurement and discusses potential applications. http://radiology.rsnajnls.org/cgi/content/full/246/1/241/DC1.


Primary orbital chronic granulomatous reaction to deep staphylococcal infection due to trauma in immunocompetent.
A 53-year-old man presented with a slow-growing mass in the right orbit and temporal area of 1 year duration following a road traffic accident. Examination revealed a nodular fungating mass involving the orbit, right temporal area with multiple discharging sinuses causing proptosis and restricted ocular motility. The patient denied light perception in the right eye. There was exposure keratopathy and optic atrophy. Imaging showed mass lesion of the orbit extending intracranially. Tissue biopsy and culture revealed a chronic granulomatous reaction due to deep infection with coagulase-positive staphylococci. The patient showed remarkable improvement to systemic antibiotics and anticonvulsants. Mortality is high in such cases due to intracranial extension if there is a delay in treatment. This case report summarises a rare aetiology and successful treatment of a case of chronic granulomatous reaction due to deep bacterial infection following trauma and is deemed rare as cases of primary orbital infection in immunocompetent have not been reported so far.


Changing Ecological Opportunities Facilitated the Explosive Diversification of New Caledonian Oxera (Lamiaceae).
Phylogenies recurrently demonstrate that oceanic island systems have been home to rapid clade diversification and adaptive radiations. The existence of adaptive radiations posits a central role of natural selection causing ecological divergence and speciation, and some plant radiations have been highlighted as paradigmatic examples of such radiations. However, neutral processes may also drive speciation during clade radiations, with ecological divergence occurring following speciation. Here, we document an exceptionally rapid and unique radiation of Lamiaceae within the New Caledonian biodiversity hotspot. Specifically, we investigated various biological, ecological, and geographical drivers of species diversification within the genus Oxera. We found that Oxera underwent an initial process of rapid cladogenesis likely triggered by a dramatic period of aridity during the early Pliocene. This early diversification of Oxera was associated with an important phase of ecological diversification triggered by significant shifts of pollination syndromes, dispersal modes, and life forms. Finally, recent diversification of Oxera appears to have been further driven by the interplay of allopatry and habitat shifts likely related to climatic oscillations. This suggests that Oxera could be regarded as an adaptive radiation at an early evolutionary stage that has been obscured by more recent joint habitat diversification and neutral geographical processes. Diversification within Oxera has perhaps been triggered by varied ecological and biological drivers acting in a leapfrog pattern, but geographic processes may have been an equally important driver. We suspect that strictly adaptive radiations may be rare in plants and that most events of rapid clade diversification may have involved a mixture of geographical and ecological divergence.


Identification of PTPN23 as a novel regulator of cell invasion in mammary epithelial cells from a loss-of-function screen of the 'PTP-ome'.
We used an RNAi-mediated loss-of-function screen to study systematically the role of the protein tyrosine phosphatase (PTP) superfamily of enzymes in mammary epithelial cell motility in the absence or presence of the oncoprotein tyrosine kinase ERBB2. We report that although shRNAs directed against most of the PTP family were without effect, suppression of three PTPs-PRPN23, PTPRG, and PTPRR-enhanced cell motility. Furthermore, we found that suppression of PTPN23, but not PTPRG or PTPRR, induced cell invasion. Suppression of PTPN23 increased E-cadherin internalization, impaired early endosome trafficking of E-cadherin, induced the expression of mesenchymal proteins, and caused cell scattering. The activity of SRC and β-catenin was elevated when PTPN23 was suppressed. Moreover, we identified SRC, E-cadherin, and β-catenin as direct substrates of PTPN23. Inhibition of SRC with the small molecular inhibitor SU6656 blocked the effects of PTPN23 depletion. These findings suggest that loss of PTPN23 may increase the activity of SRC and the phosphorylation status of the E-cadherin/β-catenin signaling complex to promote tumor growth and invasive behavior in breast cancer. In addition, our studies highlight functional specificity among PTPs and reveal new roles for PTPs in mammary epithelial cell biology.


A role for Alström syndrome protein, alms1, in kidney ciliogenesis and cellular quiescence.
Premature truncation alleles in the ALMS1 gene are a frequent cause of human Alström syndrome. Alström syndrome is a rare disorder characterized by early obesity and sensory impairment, symptoms shared with other genetic diseases affecting proteins of the primary cilium. ALMS1 localizes to centrosomes and ciliary basal bodies, but truncation mutations in Alms1/ALMS1 do not preclude formation of cilia. Here, we show that in vitro knockdown of Alms1 in mice causes stunted cilia on kidney epithelial cells and prevents these cells from increasing calcium influx in response to mechanical stimuli. The stunted-cilium phenotype can be rescued with a 5' fragment of the Alms1 cDNA, which resembles disease-associated alleles. In a mouse model of Alström syndrome, Alms1 protein can be stably expressed from the mutant allele and is required for cilia formation in primary cells. Aged mice developed specific loss of cilia from the kidney proximal tubules, which is associated with foci of apoptosis or proliferation. As renal failure is a common cause of mortality in Alström syndrome patients, we conclude that this disease should be considered as a further example of the class of renal ciliopathies: wild-type or mutant alleles of the Alström syndrome gene can support normal kidney ciliogenesis in vitro and in vivo, but mutant alleles are associated with age-dependent loss of kidney primary cilia.


Effect of an electronic nicotine delivery device (e cigarette) on desire to smoke and withdrawal, user preferences and nicotine delivery: randomised cross-over trial.
To measure the short-term effects of an electronic nicotine delivery device ("e cigarette", ENDD) on desire to smoke, withdrawal symptoms, acceptability, pharmacokinetic properties and adverse effects. Single blind randomised repeated measures cross-over trial of the Ruyan V8 ENDD. University research centre in Auckland, New Zealand. 40 adult dependent smokers of 10 or more cigarettes per day. Participants were randomised to use ENDDs containing 16 mg nicotine or 0 mg capsules, Nicorette nicotine inhalator or their usual cigarette on each of four study days 3 days apart, with overnight smoking abstinence before use of each product. The primary outcome was change in desire to smoke, measured as "area under the curve" on an 11-point visual analogue scale before and at intervals over 1 h of use. Secondary outcomes included withdrawal symptoms, acceptability and adverse events. In nine participants, serum nicotine levels were also measured. Over 60 min, participants using 16 mg ENDD recorded 0.82 units less desire to smoke than the placebo ENDD (p=0.006). No difference in desire to smoke was found between 16 mg ENDD and inhalator. ENDDs were more pleasant to use than inhalator (p=0.016) and produced less irritation of mouth and throat (p<0.001). On average, the ENDD increased serum nicotine to a peak of 1.3 mg/ml in 19.6 min, the inhalator to 2.1 ng/ml in 32 min and cigarettes to 13.4 ng/ml in 14.3 min. The 16 mg Ruyan V8 ENDD alleviated desire to smoke after overnight abstinence, was well tolerated and had a pharmacokinetic profile more like the Nicorette inhalator than a tobacco cigarette. Evaluation of the ENDD for longer-term safety, potential for long-term use and efficacy as a cessation aid is needed. Trial registration No.12607000587404, Australia and New Zealand Clinical Trials Register.


Visuomotor coordination in reaching and locomotion.
Locomotion and reaching have traditionally been regarded as separate motor activities. In fact, they may be closely connected both from an evolutionary and a neurophysiological viewpoint. Reaching seems to have evolved from the neural systems responsible for the active and precise positioning of the limb during locomotion; moreover, it seems to be organized in the spinal cord. The motor cortex and its corticospinal outflow are preferentially engaged when precise positioning of the limb is needed during locomotion and are also involved during reaching and active positioning of the hand near objects of interest. All of these motor activities require visuomotor coordination, and it is this coordination that could be achieved by the motor cortex and interconnected parietal and cerebellar areas.


Study Protocol for a Randomized, Double-Blind, Community-Based Efficacy Trial of Various Doses of Zinc in Micronutrient Powders or Tablets in Young Bangladeshi Children.
Zinc is essential to supporting growth in young children especially for tissues undergoing rapid cellular differentiation and turnover, such as those in the immune system and gastrointestinal tract. Therapeutic zinc supplementation has been initiated in low-income countries as part of diarrhea treatment programs to support these needs for young children, but the effects of preventive supplemental zinc as a tablet or as a multiple micronutrient powder (MNP) on child growth and diarrheal disease are mixed and pose programmatic uncertainties. Thus, a randomized, double-blind community-based efficacy trial of five different doses, forms, and frequencies of preventive zinc supplementation vs. a placebo was designed for a study in children aged 9⁻11 months in an urban community in Dhaka, Bangladesh. The primary outcomes of this 24-week study are incidence of diarrheal disease and linear growth. Study workers will conduct in-home morbidity checks twice weekly; anthropometry will be measured at baseline, 12 weeks and 24 weeks. Serum zinc and other related biomarkers will be measured in a subsample along with an estimate of the exchangeable zinc pool size using stable isotope techniques in a subgroup. Therapeutic zinc will be provided as part of diarrhea treatment, in accordance with Bangladesh's national policy. Therefore, the proposed study will determine the additional benefit of a preventive zinc supplementation intervention. The protocol has been approved by the Institutional Review Boards (IRBs) of icddr,b and Children's Hospital Oakland Research Institute (CHORI). The IRB review process is underway at the University of Colorado Denver as well.


Anionic cyclometallated Pt(ii) square-planar complexes: new sets of highly luminescent compounds.
Two series of novel NBu4+ salts of anionic cyclometallated Pt(ii) complexes were synthesized and fully characterized. These highly luminescent compounds (NBu4[(C^N)Pt(O^N)] and NBu4[(C^N)Pt(O^O)]) are incorporated as testing examples of cyclometallating ligands 2-phenylpyridine (PhPy), 2-(2,4-difluorophenyl)-pyridine (F2PhPy) and 2-thienylpyridine (ThPy), and a benzo[h]quinoline (Bzq) fragment. All complexes display a square-planar coordination sphere, wherein the "(C^N)Pt" fragment is completed either by an O^N orotate (Ort) or an O^O tetrabromocatecholate (Cat) ligand. The HOMO and LUMO levels of all complexes were estimated by cyclic voltammetry and a comprehensive electrochemical and photophysical study was performed. The new complexes are emissive in solution at 298 K and the NBu4[(ThPy)Pt(Ort)] complex displays good photosensitizing properties (Φ = 28% in deaerated solution vs. Φ = 1.4% in the presence of O2). Both series of NBu4[(C^N)Pt(Ort)] and NBu4[(C^N)Pt(Cat)] complexes are highly luminescent in the solid state (emission quantum yields from 10 to 85%). Remarkably, the square-planar Pt(ii) anionic complexes showed an important increase in luminescence quantum yields on changing from the dilute solution to the solid state (the most significant from 0.13% to 85% for the NBu4[(PhPy)Pt(Ort)] complex, an ideal candidate as an active species for LEECs).


Detection of gastric carcinoma-associated antigen MG7-Ag in human sera using surface plasmon resonance sensor.
MG7-Ag is a kind of gastric cancer-specific tumor-associated antigen and has been investigated to serve as a marker of gastric cancer for early diagnosis. Surface plasmon resonance (SPR) sensor was used for the detection of MG7-Ag in the sera of gastric cancer patients to develop an innovative, simple and rapid assay method for early diagnosis. The specific monoclonal MG7 antibodies were used as capture and detection receptors which were immobilized on the surface of SPR sensor chips for MG7-Ag identification in the human sera. The measurements include 9 cases of gastric cancer patients and 2 cases of healthy blood donors and a MKN45 cancer cell lysate solution sample for positive control. The binding of MG7-Ag onto the sensor surface was observed from SPR spectra. The sera of most gastric cancer patients revealed much higher expression level of MG7-Ag than healthy human sera did in SPR measurement. The initial results demonstrate that the SPR biosensor has the potential for its application in the early diagnosis of gastric cancer. However, more tests need to be done to confirm the detection limitation and the criterion for cancer risk evaluation in early diagnosis.


Transcriptional repressor germ cell-less (GCL) and barrier to autointegration factor (BAF) compete for binding to emerin in vitro.
Emerin belongs to the "LEM domain" family of nuclear proteins, which contain a characteristic approximately 40-residue LEM motif. The LEM domain mediates direct binding to barrier to autointegration factor (BAF), a conserved 10-kDa chromatin protein essential for embryogenesis in Caenorhabditis elegans. In mammalian cells, BAF recruits emerin to chromatin during nuclear assembly. BAF also mediates chromatin decondensation during nuclear assembly. The LEM domain and central region of emerin are essential for binding to BAF and lamin A, respectively. However, two other conserved regions of emerin lacked ascribed functions, suggesting that emerin could have additional partners. We discovered that these "unascribed" domains of emerin mediate direct binding to a transcriptional repressor, germ cell-less (GCL). GCL co-immunoprecipitates with emerin from HeLa cells. We determined the binding affinities of emerin for GCL, BAF, and lamin A and analyzed their oligomeric interactions. We showed that emerin forms stable complexes with either lamin A plus GCL or lamin A plus BAF. Importantly, BAF competed with GCL for binding to emerin in vitro, predicting that emerin can form at least two distinct types of complexes in vivo. Loss of emerin causes Emery-Dreifuss muscular dystrophy, a tissue-specific inherited disease that affects skeletal muscles, major tendons, and the cardiac conduction system. Although GCL alone cannot explain the disease mechanism, our results strongly support gene expression models for Emery-Dreifuss muscular dystrophy by showing that emerin binds directly to a transcriptional repressor, GCL, and by suggesting that emerin-repressor complexes might be regulated by BAF. Biochemical roles for emerin in gene expression are discussed.


Effects of Pre-Processing Hot-Water Treatment on Aroma Relevant VOCs of Fresh-Cut Apple Slices Stored in Sugar Syrup.
In practice, fresh-cut fruit and fruit salads are currently stored submerged in sugar syrup (approx. 20%) to prevent browning, to slow down physiological processes and to extend shelf life. To minimize browning and microbial spoilage, slices may also be dipped in a citric acid/ascorbic acid solution for 5 min before storage in sugar syrup. To prevent the use of chemicals in organic production, short-term (30 s) hot-water treatment (sHWT) may be an alternative for gentle sanitation. Currently, profound knowledge on the impact of both sugar solution and sHWT on aroma and physiological properties of immersed fresh-cuts is lacking. Aroma is a very important aspect of fruit quality and generated by a great variety of volatile organic compounds (VOCs). Thus, potential interactive effects of sHWT and sugar syrup storage on quality of fresh-cut apple slices were evaluated, focusing on processing-induced changes in VOCs profiles. Intact 'Braeburn' apples were sHW-treated at 55 °C and 65 °C for 30 s, sliced, partially treated with a commercial ascorbic/citric acid solution and slices stored in sugar syrup at 4 °C up to 13 d. Volatile emission, respiration and ethylene release were measured on storage days 5, 10 and 13. The impact of sHWT on VOCs was low while immersion and storage in sugar syrup had a much higher influence on aroma. sHWT did not negatively affect aroma quality of products and may replace acid dipping.


A study of proline metabolism in canola (Brassica napus L.) seedlings under salt stress.
Expression analysis of crop plants has improved our knowledge about the veiled underlying mechanisms for salt tolerance. In order to observe the time course effects of salinity stress on gene expression for enzymes regulating proline metabolism, we comparatively analyzed the expression of specific genes for proline metabolism in root and shoot tissues of salt-tolerant (cv. Dunkled) and salt-sensitive (cv. Cyclone) canola (Brassica napus L.) cultivars through reverse-transcriptase polymerase chain reaction (RT-PCR); following the NaCl treatment for various durations. Both lines showed an increase in ∆¹-pyrroline-5-carboxylate synthase1 (P5CS1) gene expression after induction of salt stress with enhanced expression in the root tissue of the tolerant line, while maximum expression was noted in the shoot tissues of the sensitive line. We observed a much reduced proline dehydrogenase (PDH) expression in both the root and shoot tissues of both canola lines, with more marked reduction of PDH expression in the shoot tissues than that in the root ones. To confirm the increase in P5CS1 gene expression, total proline content was also measured in the root and shoot tissues of both the canola lines. The root tissues of canola sensitive line showed a gradually increasing proline concentration pattern with regular increase in salinity treatment, while an increase in proline concentration in the tolerant line was noted at 24 h post salinity treatment after a sudden decrease at 6 h and 12 h of salt treatment. A gradually increasing concentration of free proline content was found in shoot tissues of the tolerant canola line though a remarkable increase in proline concentration was noted in the sensitive canola line at 24 h post salinity treatment, indicating the initiation of proline biosynthesis process in that tissue of sensitive canola.


A prospective assessment of the natural course of the exocrine pancreatic function in patients with a pancreatic head tumor.
In cancer of the pancreatic head region, exocrine insufficiency is a well-known complication, leading to steatorrhea, weight loss, and malnutrition. Its presence is frequently overlooked, however, because the primary attention is focused on cancer treatment. To date, the risk of developing exocrine insufficiency is unspecified. Therefore, we assessed this function in patients with tumors of the pancreatic head, distal common bile duct, or ampulla of Vater. Between March 2010 and August 2012, we prospectively included patients diagnosed with cancer of the pancreatic head region at our tertiary center. To preclude the effect of a resection, we excluded operated patients. Each month, the exocrine function was determined with a fecal elastase test. Furthermore, endocrine function, steatorrhea-related symptoms, and body weight were evaluated. Patients were followed for 6 months, or until death. Thirty-two patients were included. The tumor was located in the pancreas in 75%, in the bile duct in 16%, and in the ampullary region in 9%, with a median size of 2.5 cm. At diagnosis, the prevalence of exocrine insufficiency was 66%, which increased to 92% after a median follow-up of 2 months (interquartile range, 1 to 4 mo). Most patients with cancer of the pancreatic head region were already exocrine insufficient at diagnosis, and within several months, this function was impaired in almost all cases. Given this high prevalence, physicians should be focused on diagnosing and treating exocrine insufficiency, to optimize the nutritional status and physical condition, especially for those patients undergoing palliative chemotherapy and/or radiotherapy.


Regulation of Nematostella neural progenitors by SoxB, Notch and bHLH genes.
Notch signalling, SoxB and Group A bHLH 'proneural' genes are conserved regulators of the neurogenic program in many bilaterians. However, the ancestry of their functions and interactions is not well understood. We address this question in the sea anemone Nematostella vectensis, a representative of the Cnidaria, the sister clade to the Bilateria. It has previously been found that the SoxB orthologue NvSoxB(2) is expressed in neural progenitor cells (NPCs) in Nematostella and promotes the development of both neurons and nematocytes, whereas Notch signalling has been implicated in the negative regulation of neurons and the positive regulation of nematocytes. Here, we clarify the role of Notch by reporting that inhibition of Notch signalling increases the numbers of both neurons and nematocytes, as well as increasing the number of NvSoxB(2)-expressing cells. This suggests that Notch restricts neurogenesis by limiting the generation of NPCs. We then characterise NvAth-like (Atonal/Neurogenin family) as a positive regulator of neurogenesis that is co-expressed with NvSoxB(2) in a subset of dividing NPCs, while we find that NvAshA (Achaete-scute family) and NvSoxB(2) are co-expressed in non-dividing cells only. Reciprocal knockdown experiments reveal a mutual requirement for NvSoxB(2) and NvAth-like in neural differentiation; however, the primary expression of each gene is independent of the other. Together, these data demonstrate that Notch signalling and NvSoxB(2) regulate Nematostella neural progenitors via parallel yet interacting mechanisms; with different aspects of these interactions being shared with Drosophila and/or vertebrate neurogenesis.


Is your enema really necessary?
Two hundred and seventy-four women admitted for delivery of singleton infants were studied for the effects of a preparatory enema on faecal contamination, duration of labour, and the incidence of infection in the newborn. Altogether 149 of the women were given an enema (controls) and 125 were not. The two groups showed no significant difference in the degree of faecal contamination during the first and second stages of labour, and the incidences of gross contamination were similar. Contamination after an enema was especially difficult to control, since it was more likely to be fluid. Seven neonates in each group showed evidence of infection, bowel organisms being isolated from four in the no-enema group and two in the control group. Durations of labour, though not strictly comparable, were similar in the two groups. The findings suggest that when preparing for normal labour the enema should be reserved for women who have not had their bowels open in the past 24 hours and have an obviously loaded rectum on initial pelvic examination.


Feeding lactating primiparous sows to establish three divergent metabolic states: I. Associated endocrine changes and postweaning reproductive performance.
We investigated effects of different metabolic states on reproductive performance in lactating, primiparous sows. Sows were fed ad libitum (AL; n = 12), alimentated via a gastric cannula to 125% of AL feed intake (SA; n = 8), or restricted (R; n = 9) to 50% of AL from d 1 to 28 of lactation. At weaning, all sows were fed 2.5x maintenance energy requirements until standing heat and then fed twice maintenance energy requirement until slaughter. Sow weight, backfat, and litter weights were recorded weekly. After weaning, sows were tested twice daily for the onset of estrus and inseminated twice using pooled semen. At d 28 of gestation, sows were slaughtered, and the reproductive tracts were recovered to determine ovulation rate and embryo survival. Intensive blood sampling was performed before and after weaning for 12-h periods to characterize changes in plasma LH, insulin, and IGF-I. After weaning, additional samples were taken to monitor changes in LH and progesterone. Insulin and IGF-I were determined at standing heat. During lactation, AL and R sows lost, whereas SA sows gained, body weight and backfat (P < .001). Litter growth rates did not differ among treatments. Although plasma insulin was not different among treatments, plasma IGF-I concentration was lower (P < .001) in R sows. Mean LH and pulse frequency before (P < .03 and P < .06, respectively) and after (P < .001; for both) weaning were lower in R than in AL or SA sows. After weaning, SA sows lost more weight (P < .01) and backfat (P < .01) and ate less feed (P < .001) than AL or R sows. At standing heat, no differences in plasma IGF-I or insulin were observed, although energy balance for SA sows was lower (P < .01) than for AL or R sows. Weaning-to-estrus interval was extended (P < .02) in R sows. We observed no treatment difference in ovulation rate or embryo survival. Our results demonstrate that making sows anabolic during lactation did not ameliorate the negative impact of the suckling stimulus or improve fertility after weaning.


Hardware-Intrinsic Multi-Layer Security: A New Frontier for 5G Enabled IIoT.
The introduction of 5G communication capabilities presents additional challenges for the development of products and services that can fully exploit the opportunities offered by high bandwidth, low latency networking. This is particularly relevant to an emerging interest in the Industrial Internet of Things (IIoT), which is a foundation stone of recent technological revolutions such as Digital Manufacturing. A crucial aspect of this is to securely authenticate complex transactions between IIoT devices, whilst marshalling adversarial requests for system authorisation, without the need for a centralised authentication mechanism which cannot scale to the size needed. In this article we combine Physically Unclonable Function (PUF) hardware (using Field Programmable Gate Arrays-FPGAs), together with a multi-layer approach to cloud computing from the National Institute of Standards and Technology (NIST). Through this, we demonstrate an approach to facilitate the development of improved multi-layer authentication mechanisms. We extend prior work to utilise hardware security primitives for adversarial trojan detection, which is inspired by a biological approach to parameter analysis. This approach is an effective demonstration of attack prevention, both from internal and external adversaries. The security is further hardened through observation of the device parameters of connected IIoT equipment. We demonstrate that the proposed architecture can service a significantly high load of device authentication requests using a multi-layer architecture in an arbitrarily acceptable time of less than 1 second.


Model of a tunable hybrid Tamm mode-liquid crystal device.
A concept of an easily tunable device based on hybrid Tamm modes is proposed. The device can be controlled using a high-sensitivity chiral liquid crystal serving as a mirror. The coupling of the chiral optical Tamm state with the Tamm plasmons is predicted. The Tamm plasmons are excited at different frequencies for the orthogonal linear polarizations, while the chiral Tamm state is excited at only one frequency. The properties of the proposed model are analytically and numerically calculated. The possibility of creating a two- and three-mode laser with tunable characteristics on the basis of the proposed model is discussed.


Informed consent for genetic research.
Genetic research often utilizes or generates information that is potentially sensitive to individuals, families, or communities. For these reasons, genetic research may warrant additional scrutiny from investigators and governmental regulators, compared to other types of biomedical research. The informed consent process should address the range of social and psychological issues that may arise in genetic research. This paper addresses a number of these issues, including recruitment of participants, disclosure of results, psychological impact of results, insurance and employment discrimination, community engagement, consent for tissue banking, and intellectual property issues. Points of consideration are offered to assist in the development of protocols and consent processes in light of contemporary debates on a number of these issues.


Risk factors for lymphoproliferative disorders after allogeneic hematopoietic cell transplantation.
We evaluated 26 901 patients who underwent allogeneic hematopoietic cell transplantation (HCT) at 271 centers worldwide to define patterns of posttransplantation lymphoproliferative disorders (PTLDs). PTLDs developed in 127 recipients, with 105 (83%) cases occurring within 1 year after transplantation. In multivariate analyses, we confirmed that PTLD risks were strongly associated (P < .001) with T-cell depletion of the donor marrow, antithymocyte globulin (ATG) use, and unrelated or HLA-mismatched grafts (URD/HLA mismatch). Significant associations were also confirmed for acute and chronic graft-versus-host disease. The increased risk associated with URD/HLA-mismatched donors (RR = 3.8) was limited to patients with T-cell depletion or ATG use (P = .004). New findings were elevated risks for age 50 years or older at transplantation (RR = 5.1; P < .001) and second transplantation (RR = 3.5; P < .001). Lower risks were found for T-cell depletion methods that remove both T and B cells (alemtuzumab and elutriation, RR = 3.1; P = .025) compared with other methods (RR = 9.4; P = .005 for difference). The cumulative incidence of PTLDs was low (0.2%) among 21 686 patients with no major risk factors, but increased to 1.1%, 3.6%, and 8.1% with 1, 2, and more than 3 major risk factors, respectively. Our findings identify subgroups of patients who underwent allogeneic HCT at elevated risk of PTLDs for whom prospective monitoring of Epstein-Barr virus activation and early treatment intervention may be particularly beneficial.


The Italian Consensus Conference on Diagnostic Criteria for Myelofibrosis with Myeloid Metaplasia.
The purpose of this work was to develop a definition of myelofibrosis with myeloid metaplasia (MMM) using diagnostic criteria that would remain valid within the set of patients with chronic myeloproliferative disorders or myelodysplastic syndromes. A list of 12 names for the disease and 37 diagnostic criteria were proposed to a Consensus Panel of 12 Italian experts who ranked them in order so as to identify a core set of criteria. The Panel was then asked to score the diagnosis of 46 patient profiles as appropriate or not appropriate for MMM. Using the experts' consensus as the gold standard, the performance of 90 possible definitions of the disease obtained through the core set was evaluated. 'Myelofibrosis with myeloid metaplasia' ranked as the preferred name of the disease. Necessary criteria consisted of 'diffuse bone marrow fibrosis' and 'absence of Philadelphia chromosome or BCR-ABL rearrangement in peripheral blood cells'. The six optional criteria in the core set consisted of: splenomegaly of any grade; anisopoikilocytosis with tear-drop erythrocytes; the presence of circulating immature myeloid cells; the presence of circulating erythroblasts: the presence of clusters of megakaryoblasts and anomalous megakaryocytes in bone marrow sections; myeloid metaplasia. The definition of the disease with the highest final score was as follows: necessary criteria plus any other two criteria when splenomegaly is present or any four when splenomegaly is absent. The use of this definition will help to standardize the conduct and reporting of clinical studies and should help practitioners in clinical practice.