How common is clinically inactive disease in a prospective cohort of patients with juvenile idiopathic arthritis? The importance of definition.
Many criteria for clinically inactive disease (CID) and minimal disease activity (MDA) have been proposed for juvenile idiopathic arthritis (JIA). It is not known to what degree each of these criteria overlap within a single patient cohort. This study aimed to compare the frequency of MDA and CID across different criteria in a cohort of children with JIA at 1 year following presentation. The Childhood Arthritis Prospective Study recruits children at initial presentation to paediatric or adolescent rheumatology in seven UK centres. Children recruited between October 2001 and December 2013 were included. The proportions of children with CID and MDA at 1 year were calculated using four investigator-defined and eight published composite criteria. Missing data were accounted for using multiple imputation under different assumptions. In a cohort of 1415 children and adolescents, 67% patients had no active joints at 1 year. Between 48% and 61% achieved MDA and between 25% and 38% achieved CID using published criteria. Overlap between criteria varied. Of 922 patients in MDA by either the original composite criteria, Juvenile Arthritis Disease Activity Score (JADAS) or clinical JADAS cut-offs, 68% were classified as in MDA by all 3 criteria. Similarly, 44% of 633 children with CID defined by either Wallace's preliminary criteria or the JADAS cut-off were in CID according to both criteria. In a large JIA prospective inception cohort, a majority of patients have evidence of persistent disease activity after 1 year. Published criteria to capture MDA and CID do not always identify the same groups of patients. This has significant implications when defining and applying treat-to-target strategies.


Care pathways of sepsis survivors: sequelae, mortality and use of healthcare services in France, 2015-2018.
Individuals who survive sepsis are at high risk of chronic sequelae, resulting in significant health-economic costs. Several studies have focused on aspects of healthcare pathways of sepsis survivors but comprehensive, longitudinal overview of their pathways of care are scarce. The aim of this retrospective, longitudinal cohort study is to identify sepsis survivor profiles based on their healthcare pathways and describe their healthcare consumption and costs over the 3 years following their index hospitalization. The data were extracted from the French National Hospital Discharge Database. The study population included all patients above 15 years old, with bacterial sepsis, who survived an incident hospitalization in an acute care facility in 2015. To identify survivor profiles, state sequence and clustering analyses were conducted over the year following the index hospitalization. For each profile, patient characteristics and their index hospital stay and sequelae were described, as well as use of care and its associated monetary costs, both pre- and post-sepsis. New medical (79.2%), psychological (26.9%) and cognitive (18.5%) impairments were identified post-sepsis, and 65.3% of survivors were rehospitalized in acute care. Cumulative mortality reached 36.6% by 3 years post-sepsis. The total medical cost increased by 856 million € in the year post-sepsis. Five patient clusters were identified: home (65.6% of patients), early death (12.9%), late death (6.8%), short-term rehabilitation (11.3%) and long-term rehabilitation (3.3%). Survivors with early and late death clusters had high rates of cancer and primary bacteremia and experienced more hospital-at-home care post-sepsis. Survivors in short- or long-term rehabilitation clusters were older, with higher percentage of septic shock than those coming back home, and had high rates of multiple site infections and higher rates of new psychological and cognitive impairment. Over three years post-sepsis, different profiles of sepsis survivors were identified with different mortality rates, sequels and healthcare services usage and cost. This study confirmed the importance of sepsis burden and suggests that strategies of post-discharge care, in accordance with patient profile, should be further tested in order to reduce sepsis burden.


Direct mapping of bending and torsional dynamics in individual nanostructures.
Investigating coherent acoustic vibrations in nanostructured materials provides fundamental insights into optomechanical responses and microscopic energy flow. Extensive measurements of vibrational dynamics have been performed for a wide variety of nanoparticles and nanoparticle assemblies. However, virtually all of them show that only the dilation modes are launched after laser excitations, and the acoustic bending and torsional motions, which are commonly observed in photoexcited chemical bonds, are absent. Unambiguous identification and refined characterization of these "missing" modes have been a long-standing issue. In this report, we investigated the acoustic vibrational dynamics of individual Au nanoprisms on free-standing graphene substrates using an ultrafast high-sensitivity dark-field imaging approach in four-dimensional transmission electron microscopy. Following optical excitations, we observed low-frequency multiple-mode oscillations and higher superposition amplitudes at nanoprism corners and edges on the subnanoparticle level. In combination with finite-element simulations, we determined that these vibrational modes correspond to out-of-plane bending and torsional motions, superimposed by an overall tilting effect of the nanoprisms. The launch and relaxation processes of these modes are highly pertinent to substrate effects and nanoparticle geometries. These findings contribute to the fundamental understanding about acoustic dynamics of individual nanostructures and their interaction with substrates.


Influence of calcium on guanosine 3',5'-cyclic monophosphate levels in frog rod outer segments.
In the presence of 10(-9) M calcium, rod outer segments freshly detached from dark-adapted frog retinas contain between 0.01 and 0.02 moles of guanosine 3',5'-cyclic monophosphate (cyclic GMP) per mole of rhodopsin. The dark level of cyclic GMP is reduced approximately 50% by illumination that bleaches 5 x 10(5) rhodopsin molecules/outer segments. The dark levels of cyclic GMP also can be suppressed to approximately 0.007 mol/mol of rhodopsin by increasing the concentration of calcium from 10(-9) M to 2 x 10(-9) M, and they remain at this level as calcium concentration is raised to 10(-3) M. The final level to which illumination reduces cyclic GMP in unaffected by the calcium concentration between 10(-9) and 10(-3) M. The maximal light-induced decrease in cyclic GMP occurs within 1 s from the onset of illumination at all calcium concentrations. The magnitude and time-course of the light-induced decrease in cyclic GMP measured in these experiments are comparable to values obtained previously (Woodruff et al. 1977. J. Gen. Physiol. 69:677-679; Woodruff and Bownds. 1979. J. Gen. Physiol. 73:629-653). The data are consistent with a role for cyclic GMP in visual transduction irrespective of the calcium concentration.


Study of resistive switching behavior in HfO2 nanocrystals synthesized via a low temperature hydrothermal method.
The resistive switching property in HfO2 have attracted increasing interest in recent years. In this work, amorphous HfO2 nanocrystals are synthesized by a facile hydrothermal method. Then, the as-synthesized nanocrystals are rapid thermal annealed in different atmospheres for improving the crystal quality, and monoclinic phase is determined as the main crystal structure of the annealed HfO2. Subsequently, metal-insulator-metal (MIM) structure devices based on HfO2 samples are fabricated. Electrical measurement indicates that 700 oC annealing processes in Air and Ar environments can slightly improve the bipolar resistive switching and retention behaviors. Higher annealed temperature (900 oC) will further improve the crystal quality of HfO2, while the resistive switching and retention behaviors of the devices continuously attenuate, which can be ascribed to the reduction of the conductive filaments induced by defects.


Isoflavones inhibit proliferation of ovarian cancer cells in vitro via an estrogen receptor-dependent pathway.
Incidence rates of ovarian cancer remain lowest in Asian nations, which consume diets rich in soy products, whereas they remain among the highest in the United States and other Western nations, which consume low amounts of soy foods. The hypothesis of this study is that soy-derived isoflavones inhibit the proliferation of ovarian cancer cells in vitro by regulating cytokine synthesis. Cell proliferation was evaluated by bromodeoxyuridine and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. DNA synthesis of Caov-3 and NIH:OVCAR-3, two ovarian cancer cell lines, was significantly inhibited by genistein or daidzein at dietarily relevant concentrations (10(-8)-10(-10) M). Also, the number of viable cells was significantly lower (45-75%) in all isoflavone-treated groups than in the control group (P < 0.01). The addition of ICI-182780, an estrogen antagonist, blocked these inhibitory effects. In addition, interleukin-6 synthesis by these two cell lines was inhibited by genistein or daidzein; production was decreased by approximately 20% compared with the control group (P < 0.05). In contrast, transforming growth factor-beta 1 production in ovarian cancer cells incubated with genistein or daidzein was significantly greater, i.e., by approximately 30%, than in the control group (P < 0.05). Addition of ICI-182780 also neutralized the effects of isoflavones on the production of these two cytokines by ovarian cancer cells. In summary, genistein and daidzein independently modify cytokine production and reduce ovarian cancer cell proliferation via, at least in part, an estrogen receptor-dependent pathway.


Association rule mining of cellular responses induced by metal and metal oxide nanoparticles.
Relationships among fourteen different biological responses (including ten signaling pathway activities and four cytotoxicity effects) of murine macrophage (RAW264.7) and bronchial epithelial (BEAS-2B) cells exposed to six metal and metal oxide nanoparticles (NPs) were analyzed using both statistical and data mining approaches. Both the pathway activities and cytotoxicity effects were assessed using high-throughput screening (HTS) over an exposure period of up to 24 h and concentration range of 0.39-200 mg L(-1). HTS data were processed by outlier removal, normalization, and hit-identification (for significantly regulated cellular responses) to arrive at reliable multiparametric bioactivity profiles for the NPs. Association rule mining was then applied to the bioactivity profiles followed by a pruning process to remove redundant rules. The non-redundant association rules indicated that "significant regulation" of one or more cellular responses implies regulation of other (associated) cellular response types. Pairwise correlation analysis (via Pearson's χ(2) test) and self-organizing map clustering of the different cellular response types indicated consistency with the identified non-redundant association rules. Furthermore, in order to explore the potential use of association rules as a tool for data-driven hypothesis generation, specific pathway activity experiments were carried out for ZnO NPs. The experimental results confirmed the association rule identified for the p53 pathway and mitochondrial superoxide levels (via MitoSox reagent) and further revealed that blocking of the transcriptional activity of p53 lowered the MitoSox signal. The present approach of using association rule mining for data-driven hypothesis generation has important implications for streamlining multi-parameter HTS assays, improving the understanding of NP toxicity mechanisms, and selection of endpoints for the development of nanomaterial structure-activity relationships.


Micronutrient intakes, micronutrient status and lipid profiles among young people consuming different amounts of breakfast cereals: further analysis of data from the National Diet and Nutrition Survey of Young People aged 4 to 18 years.
To examine associations between breakfast cereal consumption and the dietary habits, nutrient intakes and nutritional status of young people, considering both nutrient adequacy and safety issues (fortification). Using archived data from 1688 children in the (British) National Diet and Nutrition Survey of Young People aged 4 to 18 years, nutrient intakes and status were compared across thirds of breakfast cereal consumption (T1 to T3), adjusted for age and energy intake. Cereals provided on average 2%, 6% and 12% of energy in T1, T2 and T3, respectively, for boys; 1%, 4% and 10%, respectively, for girls. Intakes of iron, B vitamins and vitamin D were around 20-60% higher in T3 compared with T1, with significant linear relationships observed for iron, thiamin, riboflavin and folate (T1<T2<T3). After excluding low energy reporters and the unwell, 14% of girls had iron intakes below the Lower Reference Nutrient Intake and this varied fivefold between T1 and T3 (27%, 12% and 5%; P=0.0001). High consumers of breakfast cereals (T3) had better folate, vitamin B12 and riboflavin status and lower total and low-density lipoprotein cholesterol. There was also an association with thiamin and vitamin B6 status in girls. However, iron status (haemoglobin, ferritin and transferrin saturation) was not significantly different between groups, possibly due to lower meat intakes in T3. Total iron intakes were within tolerable levels (maximum of 32 mg day(-1) in one girl taking supplements). The nutritional benefits of breakfast cereals are demonstrated in status measurements as well as in nutrient intakes in this study. Concerns about excessive iron intakes from fortification appear unjustified.


CE analysis of single wood cells performing hydrolysis and preconcentration in open microchannels.
In the present work, monosaccharides from pulp samples and single wood fibers were analyzed with CE, using indirect detection due to the lack of chromophores on the monosaccharides. The hydrolysis degradation of cellulose and hemicellulose into monosaccharides was performed using TFA, either in bulk scale or in microscale. In the microscale, one single wood fiber was hydrolyzed in an open microchannel manufactured on a silicon microchip with the dimensions 50 μm × 50 μm (length 1 or 3 cm). The low monosaccharide amounts derived from a single fiber implied that a preconcentration step was necessary to increase the detectability. Thus, an electromigration preconcentration of the hydrolyzed samples was performed within the microchannel, which resulted in a significantly enhanced signal intensity of the monosaccharides. In addition to the experimental study, computer simulations were performed regarding the preconcentration step of monosaccharides. The results from these simulations correlated well with the experimental results.


Development and characterization of functionalized glyco thiolate capped gold nanoparticles for biological applications.
Glyco-gold nanoparticles (AuNPs) in aqueous dispersions were prepared by two approaches, namely direct reduction and ligand substitution methods. In the direct method, potassium salts of glyco thiols, with the general formula (C6H11O6)NH(CH2) n CH2SK (where L1, n = 1; L2, n = 2; L3, n = 3, L4, n = 4; L5, n = 5), were used as reducing and capping agents to give the glyco thiolate capped gold nanoparticles (AuNPs G1-G5); meanwhile in the ligand exchange experiments, L1-L5 and their acetylated forms (L6-L8) replaced citrate ions in citrate-capped gold nanoparticles to give additional AuNPs G6-G11. UV-visible spectroscopy, surface charge (ζ-potential,) measurements and transmission electron microscopy (TEM) were used for physical and chemical characterization of all the resultant AuNPs. The ζ-potential studies of AuNPs prepared through the direct method revealed that the surface charge is dependent on the length of the alkyl unit of (C6H11O6)NH(CH2) n CH2S- ligands. TEM images of the acetylated and non-acetylated glyco thiolate capped gold nanoparticles (AuNPs G6-G11) prepared via the ligand exchange method indicate that the size and shape of the gold nanoparticles remained the same as those of the citrate-capped gold nanoparticles used to prepare them. Selected AuNPs were tested on peripheral blood mononuclear cells (PBMCs) and the A549 cancer cell line to investigate their respective toxicity and cytotoxicity profiles. All AuNPs showed indiscriminate activity against both PBMCs and A4549 cells, although the gold nanoparticles having an acetylated glyco moiety with an amino propyl thiol linker as the ligand (G10) prepared via the citrate exchange method had better selectivity (PBMCs >59 mg mL-1 and for A549 ∼7 μg mL-1).


Utilisation of complementary and alternative medicine (CAM) practitioners within maternity care provision: results from a nationally representative cohort study of 1,835 pregnant women.
There is little known about women's concurrent use of conventional and complementary health care during pregnancy, particularly consultation patterns with complementary and alternative medicine (CAM). This study examines health service utilisation among pregnant women including consultations with obstetricians, midwives, general practitioners (GPs) and CAM practitioners. A sub-study of pregnant women (n=2445) was undertaken from the nationally-representative Australian Longitudinal Study on Women's Health (ALSWH). Women's consultations with conventional practitioners (obstetricians, GPs and midwives) and CAM practitioners for pregnancy-related health conditions were analysed. The analysis included Pearson chi-square tests to compare categorical variables. The survey was completed by 1835 women (response rate = 79.2%). A substantial number (49.4%) of respondents consulted with a CAM practitioner for pregnancy-related health conditions. Many participants consulted only with a CAM practitioner for assistance with certain conditions such as neck pain (74.6%) and sciatica (40.4%). Meanwhile, women consulted both CAM practitioners and conventional maternity health professionals (obstetricians, midwives and GPs) for back pain (61.8%) and gestational diabetes (22.2%). Women visiting a general practitioner (GP) 3-4 times for pregnancy care were more likely to consult with acupuncturists compared with those consulting a GP less often (p=<0.001, x2=20.5). Women who had more frequent visits to a midwife were more likely to have consulted with an acupuncturist (p=<0.001, x2=18.9) or a doula (p=<0.001, x2=23.2) than those visiting midwives less frequently for their pregnancy care. The results emphasise the necessity for a considered and collaborative approach to interactions between pregnant women, conventional maternity health providers and CAM practitioners to accommodate appropriate information transferral and co-ordinated maternity care. The absence of sufficient clinical evidence regarding many commonly used CAM practices during pregnancy also requires urgent attention.


Exercise for pregnant women for preventing gestational diabetes mellitus.
Gestational diabetes mellitus (GDM) affects a significant number of women each year. GDM is associated with a wide range of adverse outcomes for women and their babies. Recent observational studies have found physical activity during normal pregnancy decreases insulin resistance and therefore might help to decrease the risk of developing GDM. To assess the effects of physical exercise for pregnant women for preventing glucose intolerance or GDM. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (2 April 2012), ClinicalTrials.gov (2 April 2012) and the WOMBAT Perinatal Trials Registry (2 April 2012). Randomised and cluster-randomised trials assessing the effects of exercise for preventing pregnancy glucose intolerance or GDM. Two review authors independently assessed study eligibility, extracted data and assessed the risk of bias of included studies. We included five trials with a total of 1115 women and their babies (922 women and their babies contributed outcome data). Four of the five included trials had small sample sizes with one large trial that recruited 855 women and babies. All five included trials had a moderate risk of bias. When comparing women receiving additional exercise interventions with those having routine antenatal care, there was no significant difference in GDM incidence (three trials, 826 women, risk ratio (RR) 1.10, 95% confidence interval (CI) 0.66 to 1.84), caesarean section (two trials, 934 women, RR 1.33, 95% CI 0.97 to 1.84) or operative vaginal birth (two trials, 934 women, RR 0.83, 95% CI 0.58 to 1.17). No trial reported the infant primary outcomes prespecified in the review.None of the five included trials found significant differences in insulin sensitivity. Evidence from one single large trial suggested no significant difference in the incidence of developing pregnancy hyperglycaemia not meeting GDM diagnostic criteria, pre-eclampsia or admission to neonatal ward between the two study groups. Babies born to women receiving exercise interventions had a non-significant trend to a lower ponderal index (mean difference (MD) -0.08 gram x 100 m(3), 95% CI -0.18 to 0.02, one trial, 84 infants). No significant differences were seen between the two study groups for the outcomes of birthweight (two trials, 167 infants, MD -102.87 grams, 95% CI -235.34 to 29.60), macrosomia (two trials, 934 infants, RR 0.91, 95% CI 0.68 to 1.22), or small-for-gestational age (one trial, 84 infants, RR 1.05, 95% CI 0.25 to 4.40) or gestational age at birth (two trials, 167 infants, MD -0.04 weeks, 95% CI -0.37 to 0.29) or Apgar score less than seven at five minutes (two trials, 919 infants, RR 1.00, 95% CI 0.27 to 3.65). None of the trials reported long-term outcomes for women and their babies. No information was available on health services costs. There is limited randomised controlled trial evidence available on the effect of exercise during pregnancy for preventing pregnancy glucose intolerance or GDM. Results from three randomised trials with moderate risk of bias suggested no significant difference in GDM incidence between women receiving an additional exercise intervention and routine care.Based on the limited data currently available, conclusive evidence is not available to guide practice. Larger, well-designed randomised trials, with standardised behavioural interventions are needed to assess the effects of exercise on preventing GDM and other adverse pregnancy outcomes including large-for-gestational age and perinatal mortality. Longer-term health outcomes for both women and their babies and health service costs should be included. Several such trials are in progress. We identified another seven trials which are ongoing and we will consider these for inclusion in the next update of this review.


Tumor necrosis factor therapy and the risk of serious infection and malignancy in patients with early rheumatoid arthritis: a meta-analysis of randomized controlled trials.
To conduct a meta-analysis of the rates of serious infection and malignancy in patients with early rheumatoid arthritis (RA) who have started anti-tumor necrosis factor (anti-TNF) therapy and had not received treatment with disease-modifying antirheumatic drugs (DMARDs) or methotrexate (MTX). A systematic literature search was conducted through the summer of 2009. All studies included were randomized, double-blind, placebo-controlled trials involving patients with early RA who were started on anti-TNF therapy without prior DMARD/MTX use. Six trials met the inclusion criteria for the meta-analysis, comprising a total of 2,183 patients receiving biologic therapy and 1,236 patients receiving MTX. The data extracted were from published trials. A pooled odds ratio (OR) (determined using Mantel-Haenszel methods, with a continuity correction designed for sparse data) was calculated for serious infections (requiring hospitalization) and malignancies, comparing anti-TNF therapy to MTX control. The pooled OR for serious infections was 1.28 (95% confidence interval [95% CI] 0.82-2.00) and that for malignancies was 1.08 (95% CI 0.50-2.32). There was no significant difference in either the rate of serious infections or the rate of malignancies between the anti-TNF therapy group and the control group. Whereas other meta-analyses have shown an increased risk of serious infection and malignancy in patients receiving anti-TNF therapy, the results of the present meta-analysis show that there is not an increased risk when the patients have early disease and have not previously been treated with DMARDs and/or MTX.


Neurophysiological and morphological responses to treatment with acetyl-L-carnitine in a sciatic nerve injury model: preliminary data.
We investigated the effects of acetyl-L-carnitine (ALCAR) on the recovery of sciatic nerve injuries in rats. Sprague Dawley rats were randomized to two groups: ALCAR treated (for 14 days) and control. Each group was divided into three subgroups: distal transection, proximal transection, and grafted. Distal latencies, amplitudes, and motor nerve conduction velocities were measured. In the third month, biopsies were taken and examined under light microscopy. Electrophysiological measurements demonstrated that regeneration occurred earlier and was better in the ALCAR group, particularly in the distal transection subgroup. Better results were obtained in the distal transection subgroup in terms of axonal regeneration compared with the proximal transection and grafted subgroups because the regenerating segment was shorter. ALCAR enhanced the quality of neural recovery at the different levels and in different types of repair, and led to a decline in nerve death.


Vasomotor responses in glabrous and nonglabrous skin during sinusoidal exercise.
To test whether vasomotor responses to dynamic exercise differ in glabrous and nonglabrous human skin, we determined the phase response and amplitude response of cutaneous vascular conductance in the forearm (CVCforearm), dorsal hand (CVCdorsal hand), and palm (CVCpalm) to sinusoidal exercise. Nine healthy subjects exercised on a cycle ergometer with a constant load (35% of peak O2 uptake) for 20 min at an ambient temperature of 25 degrees C and relative humidity of 60%; for the next 40 min, they exercised with a sinusoidal load. The sinusoidal load variation ranged from 10% to 60% of peak O2 uptake over a 4-min period. Skin blood flow was monitored by laser-Doppler flowmetry. CVC was calculated from the ratio of blood flow to mean arterial pressure (MAP). During sitting rest and exercise, CVCpalm showed consistently higher value than CVCforearm and CVCdorsal hand. During sinusoidal exercise, the amplitude in CVCpalm was 7.4 times and 3.2 times greater than those in CVCforearm and CVCdorsal hand, respectively (P < 0.05). The phase difference in CVCforearm and CVCdorsal hand were smaller than that in CVCpalm (P < 0.05). These findings of significant differences in phase and amplitude of responses in CVC between glabrous and nonglabrous skin during cyclic changes of dynamic exercise load suggest functionally important differences in the reflex control of these regions of skin.


Transcranial Magnetic Stimulation Promotes Gait Training in Parkinson Disease.
To determine whether priming with 1 or 25Hz repetitive transcranial magnetic stimulation (rTMS) will enhance the benefits from treadmill training up to 3 months postintervention in people with Parkinson disease (PD), and to evaluate the underlying changes in cortical excitability. This randomized double-blind, placebo-controlled trial was conducted between October 2016 and December 2018. Fifty-one participants with PD were randomized to receive 12 sessions of rTMS (25Hz, 1Hz, or sham) followed by treadmill training. All participants were assessed at baseline and 1 day, 1 month, and 3 months postintervention. Primary outcome was fastest walking speed, and secondary outcomes were timed up-and-go test (TUG), dual-task TUG (DT-TUG), motor section of the Movement Disorder Society-Unified Parkinson's Disease Rating Scale (MDS-UPDRS-III), and electrophysiological evaluation of cortical excitability by TMS. The 1 and 25Hz rTMS groups produced a greater improvement in fastest walking speed at 1 day and 3 months postintervention than the sham group. Only the 1 and 25Hz rTMS groups sustained the improvements in TUG, and had a significant improvement in DT-TUG and MDS-UPDRS-III for up to 3 months. Behavioral improvements correlated with increased cortical silent period and short-interval intracortical inhibition in both groups receiving real rTMS. Priming with 1 and 25Hz rTMS can augment the benefits of treadmill training and lead to long-term motor improvement up to 3 months postintervention. The motor improvement at follow-up was associated with a normalization of cortical excitability, which in turn suggests an alteration of the homeostatic plasticity range. Rebalancing cortical excitability by rTMS appears critical for plasticity induction. ANN NEUROL 2020;88:933-945.


Optical gating and streaking of free electrons with sub-optical cycle precision.
The temporal resolution of ultrafast electron diffraction and microscopy experiments is currently limited by the available experimental techniques for the generation and characterization of electron bunches with single femtosecond or attosecond durations. Here, we present proof of principle experiments of an optical gating concept for free electrons via direct time-domain visualization of the sub-optical cycle energy and transverse momentum structure imprinted on the electron beam. We demonstrate a temporal resolution of 1.2±0.3 fs. The scheme is based on the synchronous interaction between electrons and the near-field mode of a dielectric nano-grating excited by a femtosecond laser pulse with an optical period duration of 6.5 fs. The sub-optical cycle resolution demonstrated here is promising for use in laser-driven streak cameras for attosecond temporal characterization of bunched particle beams as well as time-resolved experiments with free-electron beams.


Motif-VI Loop Acts as a Nucleotide Valve in the West Nile Virus NS3 Helicase.
The flavivirus NS3 helicase (NS3h), a highly conserved protein, plays a pivotal role in virus replication and thus represents a potential drug target for flavivirus pathogenesis. NS3h utilizes nucleotide triphosphate, such as ATP, for hydrolysis energy (ATPase) to translocate on single-stranded nucleic acids, which is an important step in the unwinding of double-stranded nucleic acids. The intermediate states along the ATP binding and hydrolysis cycle, as well as the conformational changes between these states, represent important yet difficult-to-identify targets for potential inhibitors. We use extensive molecular dynamics simulations of apo, ATP, ADP+P i , and ADP bound to WNV NS3h+ssRNA to model the conformational ensembles along this cycle. Energetic and structural clustering analyses on these trajectories depict a clear trend of differential enthalpic affinity of NS3h with ADP, demonstrating a probable mechanism of hydrolysis turnover regulated by the motif-VI loop (MVIL). These findings were experimentally corroborated using viral replicons encoding three mutations at the D471 position. Replication assays using these mutants demonstrated a substantial reduction in viral replication compared to the wild-type. Molecular simulations of the D471 mutants in the apo state indicate a shift in MVIL populations favoring either a closed or open 'valve' conformation, affecting ATP entry or stabilization, respectively. Combining our molecular modeling with experimental evidence highlights a conformation-dependent role for MVIL as a 'valve' for the ATP-pocket, presenting a promising target for antiviral development.


[Ruptures of the bladder. Surgical or conservative treatment].
Since the bladder is well protected by the bony pelvis, it is seldom injured. However, the initial trauma may be a major one, and associated lesions are frequent, especially in case of extraperitoneal ruptures. Retrograde cystography still remains an essential examination, for which the technique must be straightforward. This work is a review of our cases of pure bladder rupture from 1981 to 1992. Investigations, treatment, and our results are discussed, and the literature is reviewed. In the last few years, we have been favoring a conservative approach of extraperitoneal lesions.


Effects of feeding alkaline hydrogen peroxide-treated wheat straw-based diets on intake, digestion, ruminal fermentation, and production responses by mid-lactation dairy cows.
The objective of this experiment was to determine the effects of feeding different levels of alkaline hydrogen peroxide-treated wheat straw (AHP-WS) in the diet on feed intake, nutrient digestion, ruminal fermentation, and production responses in mid-lactation dairy cows. Eight Holstein cows, averaging 147 d postpartum, were used in two replications of a 4 x 4 Latin square design. Complete mixed diets consisted of 70% forage and 30% concentrate (DM basis) with various levels of AHP-WS, alfalfa haylage, and corn silage as forage sources. Treatments contained 0 (control), 20.0, 40.1, or 60.0% AHP-WS in the diet. A quadratic effect (P = .08) of AHP-WS level on DMI was noted, with values of 2.16, 22.3, 20.8, and 18.9 kg/d for the control, 20.0, 40.1, and 60.0% AHP-WS treatments, respectively. Apparent digestibilities of DM, OM, CP, and ADF were not affected (P greater than .10) by replacing haylage and corn silage with increasing amounts of AHP-WS in the diet, but there was a linear increase (P = .03) in NDF digestibility (44% for control vs 59% for the 60.0% AHP-WS diet) and a parallel decrease (P less than .05) in cell content digestibility (82 vs 70% for these two diets). Yields of milk and 4% fat-corrected milk (FCM) were decreased (quadratic; P = .0001) as the level of AHP-WS increased in the diet. The addition of AHP-WS to the diet decreased the milk fat percentage from 3.72 to 3.60% (quadratic; P = .05) and decreased milk protein percentage from 3.27 to 3.13% (linear; P = .0001). Cows fed the higher levels of AHP-WS had linear increases (P = .0001) in ruminal concentrations of total VFA (128.0 mM for control vs 136.0 mM for the 60.0% AHP-WS treatment) and molar proportion of acetate, resulting in a quadratic effect (P less than .0001) on the acetate:propionate ratio. These data indicate that feeding the 40.1 and 60.0% AHP-WS diets lowered digestible DM and OM intakes, which resulted in reduced 4% FCM yield as nutrient intakes were decreased compared with cows fed the 20.0% AHP-WS diet or the control diet containing alfalfa haylage and corn silage. Although substituting AHP-WS for haylage and corn silage increased NDF digestibility and tended to increase digestible NDF intake, milk production was depressed because digestible DMI decreased.


Anti-CEA-functionalized superparamagnetic iron oxide nanoparticles for examining colorectal tumors in vivo.
Although the biomarker carcinoembryonic antigen (CEA) is expressed in colorectal tumors, the utility of an anti-CEA-functionalized image medium is powerful for in vivo positioning of colorectal tumors. With a risk of superparamagnetic iron oxide nanoparticles (SPIONPs) that is lower for animals than other material carriers, anti-CEA-functionalized SPIONPs were synthesized in this study for labeling colorectal tumors by conducting different preoperatively and intraoperatively in vivo examinations. In magnetic resonance imaging (MRI), the image variation of colorectal tumors reached the maximum at approximately 24 h. However, because MRI requires a nonmetal environment, it was limited to preoperative imaging. With the potentiality of in vivo screening and intraoperative positioning during surgery, the scanning superconducting-quantum-interference-device biosusceptometry (SSB) was adopted, showing the favorable agreement of time-varied intensity with MRI. Furthermore, biological methodologies of different tissue staining methods and inductively coupled plasma (ICP) yielded consistent results, proving that the obtained in vivo results occurred because of targeted anti-CEA SPIONPs. This indicates that developed anti-CEA SPIONPs owe the utilities as an image medium of these in vivo methodologies.


Integrating competence and psychopathology: pathways toward a comprehensive science of adaptation in development.
This paper examines the conceptual and empirical connections between competence and psychopathology, two historically rich traditions for the study of adaptation in development, and what might be gained from their integration. Historical roots of these two traditions are reviewed, then overlaps in their definition are considered, with a focus on the ways in which judgments about competence enter into the nosology of mental disorders. DSM-IV is analyzed from the perspective of competence, and the debate about "harmful dysfunction" in defining mental disorder is discussed in relation to competence. Different models explaining the empirical associations of competence and psychopathology are delineated, and illustrative empirical evidence is provided. Potential explanations include confounded concepts and methods, symptoms undermining the effectiveness of adaptation in the environment, failures in age-salient developmental tasks leading to emotional and behavioral problems, transactional influences, shared vulnerability or risk factors producing both kinds of difficulties, and more complex models. The potential benefits of integrating competence and psychopathology as two major approaches to adaptation are discussed in regard to theory, classification of mental disorder, research, and intervention.


Antitumoral activity of allosteric inhibitors of protein kinase CK2.
Due to its physiological role into promoting cell survival and its dysregulation in most cancer cells, protein kinase CK2 is a relevant physiopathological target for development of chemical inhibitors. We report the discovery of azonaphthalene derivatives, as a new family of highly specific CK2 inhibitors. First, we demonstrated that CK2 inhibition (IC50= 0.4 µM) was highly specific, reversible and non ATP-competitive. Small Angle X-ray Scattering experiments showed that this inhibition was due to large conformational change of CK2α upon binding of these inhibitors. We showed that several compounds of the family were cell-potent CK2 inhibitors promoting cell cycle arrest of human glioblastoma U373 cells. Finally, in vitro and in vivo assays showed that these compounds could decrease U373 cell tumor mass by 83 % emphasizing their efficacy against these apoptosis-resistant tumors. In contrast, Azonaphthalene derivatives inactive on CK2 activity showed no effect in colony formation and tumor regression assays. These findings illustrate the emergence of nonclassical CK2 inhibitors and provide exciting opportunities for the development of novel allosteric CK2 inhibitors. CK2 is an emerging therapeutic target and ATP-competitive inhibitors have been identified. CK2 is endowed with specific structural features providing alternative strategies for inhibition. Azonaphthalene compounds are allosteric CK2 inhibitors showing antitumor activity. CK2 may be targeted allosterically. These inhibitors provide a foundation for a new paradigm for specific CK2 inhibition.


A strategy of designing high-entropy alloys with high-temperature shape memory effect.
Shape memory effect, the ability to recover a pre-deformed shape on heating, results from a reversible martensitic transformation between austenite and martensite phases. Here, we demonstrate a strategy of designing high-entropy alloys (HEAs) with high-temperature shape memory effect in the CrMnFeCoNi alloy system. First, we calculate the difference in Gibbs free energy between face-centered-cubic (FCC) and hexagonal-close-packed (HCP) phases, and find a substantial increase in thermodynamic equilibrium temperature between the FCC and HCP phases through composition tuning, leading to thermally- and stress-induced martensitic transformations. As a consequence, the shape recovery temperature in non-equiatomic CrMnFeCoNi alloys can be increased to 698 K, which is much higher than that of conventional shape memory alloys (SMAs) and comparable to that of B2-based multi-component SMAs containing noble metals (Pd, Pt, etc.) or refractory metals (Zr, Hf, etc.). This result opens a vast field of applications of HEAs as a novel class of cost-effective high-temperature SMAs.


Corticotropin-releasing hormone receptors mediate apoptosis via cytosolic calcium-dependent phospholipase A₂ and migration in prostate cancer cell RM-1.
Peripheral corticotropin-releasing hormone receptors (CRHRs) are G protein-coupled receptors that play different roles depending on tissue types. Previously, we discovered the mechanism of CRHR-mediated apoptosis of mouse prostate cancer cell line (RM-1) to be a change of Bcl-2:Bax ratio, and CRH was found to inhibit transforming growth factor β migration of breast cancer cells via CRHRs. In the present study, we investigated cytosolic calcium-dependent phospholipase A2 (cPLA2) bridging CRHR activations and Bcl-2:Bax ratio and the effect of CRHR activation on cell migration. Silencing of cPLA2 attenuated a CRHR1 agonist, CRH-induced apoptosis, and the decrease of the Bcl-2:Bax ratio, whereas silencing of cPLA2 aggravated CRHR2 agonist, Urocortin 2 (Ucn2)-inhibited apoptosis, and the increase of the Bcl-2:Bax ratio. CRH in a time- and concentration-dependent manner increased cPLA2 expression mainly through interleukin 1β (IL1β) upregulation. Ucn2 decreased cPLA2 expression through neither tumor necrosis factor α nor IL1β. CRH-suppressed decay of cPLA2 mRNA and Ucn2 merely suppressed its production. Overexpression of CRHR1 or CRHR2 in HEK293 cells correspondingly upregulated or downregulated cPLA2 expression after CRH or Ucn2 stimulation respectively. In addition, both CRH and Ucn2 induced migration of RM-1 cells. Our observation not only established a relationship between CRHRs and cell migration but also for the first time, to our knowledge, demonstrated that cPLA2 participates in CRHR1-induced apoptosis and CRHR2-inhibited apoptosis.


Chemical Synthesis Demonstrates That Dynamic O-Glycosylation Regulates the Folding and Functional Conformation of a Pivotal EGF12 Domain of the Human NOTCH1 Receptor.
The interaction of the human NOTCH1 receptor and its ligands is a crucial step in initiating the intracellular signal transductions, in which O-glycosylation of the extracellular EGF-like domain strongly affects multiple aspects of cell differentiation, development, and cancer biology. However, consequences of biosynthetic O-glycosylation processes in the endoplasmic reticulum (ER) and Golgi on the folding of EGF domains remain unclear. Synthetic human NOTCH1 EGF12 modules allow for new insight into the crucial roles of O-glycosylation in the folding and conformation of this pivotal domain. Here, we show for the first time that predominant O-glucosylation at Ser458 facilitates proper folding of the EGF12 domain in the presence of calcium ion, while the nonglycosylated linear EGF12 peptide affords large amounts of misfolded products (>50%) during in vitro oxidative folding. Strikingly, O-fucosylation at Thr466 prior to O-glucosylation at Ser458 totally impedes folding of EGF12 independent of calcium ion, whereas modification of the Fucα1→ moiety with β-linked GlcNAc dramatically enhances folding efficiency. In addition, we elicit that extension of the Glcβ1→ moiety with xyloses is a negative-regulation mechanism in the folding of EGF12 when synthesis of a trisaccharide (Xylα1→3Xylα1→3Glcβ1→) dominates over the posttranslational modification at Thr466. Comprehensive nuclear magnetic resonance studies of correctly folded EGF12 modules demonstrate that noncovalently bonded bridges between sugars and peptide moieties, namely sugar bridges, contribute independently to the stabilization of the antiparallel β-sheet in the ligand-binding region. Our results provide evidence that the dynamic O-glycosylation status of the EGF12 domain elaborated in the ER and Golgi strongly affects folding and trafficking of the human NOTCH1 receptor.


Addressing Veteran Homelessness to Prevent Veteran Suicides.
The U.S. Department of Veterans Affairs (VA) is shifting its focus from ending veteran homelessness to preventing veteran suicides. With supporting data, this Open Forum argues that VA homelessness services also help address veteran suicides. Analysis of a nationally representative survey of U.S. veterans in 2015 shows that veterans with a history of homelessness attempted suicide in the previous two years at a rate >5.0 times higher compared with veterans without a history of homelessness (6.9% versus 1.2%), and their rates of two-week suicidal ideation were 2.5 times higher (19.8% versus 7.4%). Because the majority of veterans who die by suicide are not engaged in VA care, VA services for the homeless that include outreach efforts to engage new veterans may be reaching some of these veterans. Thus continued federal support for VA homelessness services not only may help address homelessness but also may help prevent suicide of veterans.


Methodological issues in conducting and analyzing longitudinal observational studies in rheumatoid arthritis.
This article discusses 5 methodological issues that arise in the course of conducting longitudinal observational studies: generalizability, missing data, repeated measures on the same individual, measures taken at varying time points from symptom onset, and assessing the effect of treatment. Methods discussed include general estimating equations and propensity scores. The points are illustrated by examples from the Norfolk Arthritis Register dataset.


Intestinal transport of zinc and folic acid: a mutual inhibitory effect.
Recent observations suggest an inverse relationship between folic acid intake and zinc nutriture and indicate an interaction between folic acid and zinc at the intestinal level. To define that interaction, we designed in vivo and in vitro transport studies in which folic acid transport in the presence of zinc, as well as zinc transport in the presence of folic acid was examined. These studies show that zinc transport is significantly decreased when folate is present in the intestinal lumen. Similarly folic acid transport is significantly decreased with the presence of zinc. To determine whether this intestinal inhibition is secondary to zinc and folate-forming complexes, charcoal-binding studies were performed. These studies indicate that zinc and folate from complexes at pH 2.0, but that at pH 6.0, these complexes dissolve. Therefore, our studies suggest that under normal physiological conditions a mutual inhibition between folate and zinc exists at the site of intestinal transport.


A non-randomised experimental feasibility study into the immediate effect of three different spinal manipulative protocols on kicking speed performance in soccer players.
The most utilized soccer kicking method is the instep kicking technique. Decreased motion in spinal joint segments results in adverse biomechanical changes within in the kinematic chain. These changes may be linked to a negative impact on soccer performance. This study tested the immediate effect of lumbar spine and sacroiliac manipulation alone and in combination on the kicking speed of uninjured soccer players. This 2010 prospective, pre-post experimental, single-blinded (subject) required forty asymptomatic soccer players, from regional premier league teams, who were purposively allocated to one of four groups (based on the evaluation of the players by two blinded motion palpators). Segment dysfunction was either localized to the lumbar spine (Group 1), sacroiliac joint (Group 2), the lumbar spine and sacroiliac joint (Group 3) or not present in the sham laser group (Group 4). All players underwent a standardized warm-up before the pre-measurements. Manipulative intervention followed after which post-measurements were completed. Measurement outcomes included range of motion changes (digital inclinometer); kicking speed (Speed Trac™ Speed Sport Radar) and the subjects' perception of a change in kicking speed. SPSS version 15.0 was used to analyse the data, with repeated measures ANOVA and a p-value <0.05 (CI 95%). Lumbar spine manipulation resulted in significant range of motion increases in left and right rotation. Sacroiliac manipulation resulted in no significant changes in the lumbar range of motion. Combination manipulative interventions resulted in significant range of motion increases in lumbar extension, right rotation and right SI joint flexion. There was a significant increase in kicking speed post intervention for all three manipulative intervention groups (when compared to sham). A significant correlation was seen between Likert based-scale subjects' perception of change in kicking speed post intervention and the objective results obtained. This pilot study showed that lumbar spine manipulation combined with SI joint manipulation, resulted in an effective intervention for short-term increases in kicking speed/performance. However, the lack of an a priori analysis, a larger sample size and an unblinded outcome measures assessor requires that this study be repeated, addressing these concerns and for these outcomes to be validated.


Aquaporin-4 autoantibodies in a paraneoplastic context.
The neuromyelitis optica IgG autoantibody (NMO-IgG) is a validated biomarker for NMO and an emerging spectrum of inflammatory central nervous system-demyelinating disorders. Its antigen is the astrocytic water channel aquaporin-4; NMO-IgG has not been described in a cancer context. To report (1) neurologic and oncologic correlates for patients incidentally identified as NMO-IgG seropositive in a blinded evaluation for paraneoplastic autoantibodies and (2) the frequency of cancer in NMO-IgG-seropositive patients. Observational, retrospective case series. Neuroimmunology Laboratory and Neurology Clinical Practice, Mayo Clinic College of Medicine. From 1998 to 2007, we detected NMO-IgG in 2 patient groups: (1) 31 patients (88% female) identified incidentally among 180 000 patients evaluated for paraneoplastic autoantibodies and (2) 141 patients identified through physician-requested serological evaluation for a suspected NMO-spectrum disorder. In the first group, clinical information was available for 28 patients (90%). An NMO-spectrum disorder was diagnosed in 26 patients (93%), of whom 6 had a neoplasm (5 carcinomas [2 breast, 1 lung, 1 thymic, and 1 uterine cervical] and 1 B-cell lymphoma) and 1 had monoclonal gammopathy. In 4 patients, NMO-related symptoms followed neoplasia detection (median, 14 [range 3-18] months), and in 2 patients, symptoms preceded neoplasia detection (by 5 and 3 months). Two patients had carcinoma (1 breast and 1 lung) without neurological evidence of an NMO-spectrum disorder. In the second group, neoplasms were recorded in 7 seropositive patients (5.0%) with a clinically diagnosed NMO-spectrum disorder: 3 carcinomas (all breast), 1 thyroid Hürthle cell, 1 carcinoid, 1 pituitary somatotropinoma, and 1 B-cell lymphoma. An eighth patient had monoclonal gammopathy. Aquaporin-4-specific IgG in some cases of NMO may reflect a paraneoplastic immune response. The clinical utility of this autoantibody as a cancer marker warrants prospective investigation.


Neglected tropical diseases: infection, modeling, and control.
We survey the current state of a group of parasitic and microbial diseases called the Neglected Tropical Diseases (NTDs). These diseases currently infect a billion people, primarily in socioeconomically depressed areas of the world, are a leading cause of worldwide disability, and are responsible for approximately 534,000 deaths per year. We focus on several subcategories: protozoans, helminthes and bacterial diseases. We identify the populations most at risk from these diseases, and outline symptoms and other disease burdens. We examine the progress being made in controlling NTDs, including the current state of drug development. We also examine mathematical modeling of NTDs. While mathematical modeling is not bound by many of the strictures of access, data collection and infrastructure funding, we nevertheless demonstrate that few NTDs have received much attention from mathematical models, and that some have received no attention at all. Simple mathematical models could contribute significantly to our understanding of these diseases and the efforts required to control them, at very little cost. Further investment in prevention, treatment and awareness of NTDs is urgently warranted.


Crystal structure of a complex between the Actinomadura R39 DD-peptidase and a peptidoglycan-mimetic boronate inhibitor: interpretation of a transition state analogue in terms of catalytic mechanism.
The Actinomadura R39 DD-peptidase is a bacterial low molecular weight class C penicillin-binding protein. It has previously been shown to catalyze hydrolysis and aminolysis of small D-alanyl-D-alanine terminating peptides, especially those with a side chain that mimics the amino terminus of the stem peptide precursor to the bacterial cell wall. This paper describes the synthesis of (D-alpha-aminopimelylamino)-D-1-ethylboronic acid, designed to be a peptidoglycan-mimetic transition state analogue inhibitor of the R39 DD-peptidase. The boronate was found to be a potent inhibitor of the peptidase with a K(i) value of 32 +/- 6 nM. Since it binds some 30 times more strongly than the analogous peptide substrate, the boronate may well be a transition state analogue. A crystal structure of the inhibitory complex shows the boronate covalently bound to the nucleophilic active site Ser 49. The aminopimelyl side chain is bound into the site previously identified as specific for this moiety. One boronate oxygen is held in the oxyanion hole; the other, occupying the leaving group site of acylation or the nucleophile site of deacylation, appears to be hydrogen-bonded to the hydroxyl group of Ser 298. The Ser 49 oxygen appears to be hydrogen bonded to Lys 52. If it is assumed that this structure does resemble a high-energy tetrahedral intermediate in catalysis, it seems likely that Ser 298 participates as part of a proton transfer chain initiated by Lys 52 or Lys 410 as the primary proton donor/acceptor. The structure, therefore, supports a particular class of mechanism that employs this proton transfer device.


SnakeWRAP: a Snakemake workflow to facilitate automated processing of metagenomic data through the metaWRAP pipeline.
Generating high-quality genome assemblies of complex microbial populations from shotgun metagenomics data is often a manually intensive task involving many computational steps. SnakeWRAP is a novel tool, implemented in the Snakemake workflow language, to automate multiple metaWRAP modules. Specifically, it wraps the shell scripts provided within the original metaWRAP software, within Snakemake. This approach enables high-throughput simultaneous assembly and analysis of multiple shotgun metagenomic datasets using the robust modular metaWRAP software. We expect this advancement to be of import in institutions where high-performance computing infrastructure is available, especially in the context of big data. This software tool is publicly available at https://github.com/jkrapohl/SnakeWRAP.


Development of a novel in vitro cell model for evaluation of nanofiber mats immunogenicity.
Immunological safety of nanofibers remains poorly reported within the scientific literature and lacks specific in vitro testing models distinct from those used to test nanoparticles. To address the challenges of currently used conventional setups being described in the literature, we developed a novel in vitro model for nanofiber mats immunogenicity testing, which enables standardization of tested surface area, excludes nanofiber mat edges, and ensures stable contacts of cells with nanofibers during the experiment. The effect of nanofibers was assessed on peripheral blood mononuclear cells (PBMCs) by measuring their metabolic activity using MTS cell proliferation assay, where key performance parameters, i.e. cell number, phytohemagglutinin-L (PHA-L) concentration, incubation time and cell lysis were optimized. Repeatability of results obtained with non-activated and PHA-L-activated PBMCs in contact with differently thick polycaprolactone nanofiber mats was compared using both models. Our model provided more reproducible results with lower variability, exhibiting its higher reliability and accuracy than the conventional one. Furthermore, results showed the presence of thicker mats resulted in reduced metabolic activity and PBMC proliferation without any observed cytotoxicity, providing additional insights into their non-immunogenic characteristics. The developed model enables more accurate biological assessment that can support new guidelines for in vitro nanofiber testing and formulation.


Corticosteroids in organ preservation.
1. Glucocorticoids have been empirically employed during perfusion preservation of transplantable organs. 2. Use of corticosteroids in organ preservation is based on their pharmocologic binding of released lysosomal enzymes, as membrane stabilizers, peripheral vasodilators, and inhibitors of the inflammatory response. 3. The incidence post-transplant of ATN in cadaveric kidney preservation by pulsa-tile perfusion has been reduced from 57% to 18% when the donor was protected with large doses of M-P. 4. There is sufficient clinical and laboratory evidence to substantiate the use of large doses of M-P given to the prospective donor as a protective agent against ischemic injury in organ recovery.


Requirement of Tyr-992 and Tyr-1173 in phosphorylation of the epidermal growth factor receptor by ionizing radiation and modulation by SHP2.
The epidermal growth factor receptor (EGFR) is activated by ionizing radiation (IR) in many human carcinomas, mediating a cytoprotective response and subsequent radioresistance. The underlying molecular mechanisms remain to be understood, and we propose here a specific role for the Tyr-992 residue of EGFR and examine its regulation by the phosphatase, SHP2. The -fold increase in phosphorylation of Tyr-992 in response to IR is twice that seen with ligand (EGF) binding. Mutation of Tyr-992 blocked completely IR-induced EGFR phosphorylation and reduced activation of the downstream signaling molecule, phospholipase Cgamma. IR has previously been demonstrated to inhibit activity of protein-tyrosine phosphatases. Following protein-tyrosine phosphatase inhibition by sodium vanadate both EGFR expressing Chinese hamster ovary (CHO) and A431 exhibited up to an 8-fold increase in the basal level of Tyr-992 phosphorylation, significantly higher than that seen with Tyr-1173, Tyr-1068, and total EGFR Tyr. CHO cells expressing a SHP2 mutant also demonstrated up to an 8-fold increase in the basal level of Tyr-992 phosphorylation. In this study we show the unique association of SHP2 with EGFR in response to IR, with up to a 2.5-fold increase in the direct association of endogenous SHP2 with EGFR-wt in response to 2 gray of IR in both CHO and A431 cells. Mutation of Tyr-992 abolished this response. In conclusion we have identified several differentially activated Tyr residues, one of which is not only more sensitive to activation by IR, translating into differential activation of downstream signaling, but uniquely modulated by the phosphatase SHP2.


The characterization of macrophages and osteoclasts in tissues harvested from revised total hip prostheses.
The differentiation and maturation of macrophages and osteoclasts at the prosthetic interface in cases of implant loosening are poorly understood. Using histochemical and immunohistochemical staining methods, we compare macrophage differentiation in tissues from revised hip replacements in patients with specific clinical-radiological appearances. Periprosthetic tissues were harvested from 12 cemented acetabular and 12 cemented femoral components in 24 patients undergoing revision hip replacement. The prostheses were all radiographically and clinically loose. Six acetabular and six femoral components demonstrated radiographic ballooning osteolysis. Serial 6 microm frozen sections of the periprosthetic tissues were processed with hematoxylin and eosin for general tissue morphology, and analyzed for the presence of tartrate resistant acid phosphatase (TRAP, an osteoclast marker). Immunoperoxidase staining using monoclonal antibodies to CD68 (macrophages and osteoclasts) and CD51 (the alpha chain of the vitronectin receptor, an osteoclast marker) was also performed. Approximately 8-30% of the total cells in the tissues were positive for TRAP and the vitronectin receptor, and comprised a subset of the CD68 positive macrophages and macrophage polykaryons. However, there were no statistically significant differences between specific groups (femoral vs. acetabular, osteolysis vs. no osteolysis) for the numbers or percentages of macrophages or osteoclast-like cells. Once prosthetic loosening has occurred, few differences in the macrophage-osteoclast profile of tissues from different periprosthetic locations, with and without osteolysis, are noted. This suggests a final common biologic pathway for periprosthetic bone resorption, once implant loosening has transpired.


Tetramethylenedisulfotetramine neurotoxicity: What have we learned in the past 70 years?
Tetramethylenedisulfotetramine (tetramine, TETS, TMDT) is a seizure-producing neurotoxic chemical formed by the condensation of sulfamide and formaldehyde. Serendipitously discovered through an occupational exposure in 1949, it was promoted as a rodenticide but later banned worldwide due to its danger to human health. However, exceptional activity of the agent against rodent pests resulted in its clandestine manufacture with large numbers of inadvertent, intentional, and mass poisonings, which continue to this day. Facile synthesis, extreme potency, persistence, lack of odor, color, and taste identify it as an effective food adulterant and potential chemical agent of terror. No known antidote or targeted treatment is currently available. In this review we examine the origins of tetramethylenedisulfotetramine, from its identification as a neurotoxicant 70 years ago, through early research, to the most recent findings including the risk it poses in the post-911 world. Included is the information known regarding its in vitro pharmacology as a GABAA receptor channel antagonist, the toxic syndrome it produces in vivo, and its effect upon vulnerable populations. We also summarize the available information about potential therapeutic countermeasures and treatment strategies as well as the contribution of clinical development of TMDT poisoning to our understanding of epileptogenesis. Finally we identify gaps in our knowledge and suggest potentially fruitful directions for continued research on this dangerous, yet intriguing compound.


Cloning, expression, and cell surface localization of Paenibacillus sp. strain W-61 xylanase 5, a multidomain xylanase.
We have shown that a xylan-degrading bacterium, W-61, excretes multiple xylanases, including xylanase 5 with a molecular mass of 140 kDa. Here, we emend the previously used classification of the bacterium (i.e., Aeromonas caviae W-61) to Paenibacillus sp. strain W-61 on the basis of the nucleotide sequence of the 16S rRNA gene, and we clone and express the xyn5 gene encoding xylanase 5 (Xyn5) in Escherichia coli and study the subcellular localization of Xyn5. xyn5 encodes 1,326 amino acid residues, including a 27-amino-acid signal sequence. Sequence analysis indicated that Xyn5 comprises two family 22 carbohydrate-binding modules (CBM), a family 10 catalytic domain of glycosyl hydrolases, a family 9 CBM, a domain similar to the lysine-rich region of Clostridium thermocellum SdbA, and three S-layer-homologous (SLH) domains. Recombinant Xyn5 bound to a crystalline cellulose, Avicel PH-101, while an N-terminal 90-kDa fragment of Xyn5, which lacks the C-terminal half of the family 9 CBM, did not bind to Avicel PH-101. Xyn5 was cell bound, and the cell-bound protein was digested by exogenous trypsin to produce immunoreactive and xylanolytic fragments with molecular masses of 80 and 60 kDa. Xyn5 was exclusively distributed in the cell envelope fraction consisting of a peptidoglycan-containing layer and an associated S layer. Thus, Paenibacillus sp. strain W-61 Xyn5 is a cell surface-anchored modular xylanase possessing a functional cellulose-binding module and SLH domains. Possible cooperative action of multiple xylanases produced by strain W-61 is discussed on the basis of the modular structure of Xyn5.


Regulation of brain anandamide by acute administration of ethanol.
The endogenous cannabinoid acylethanolamide AEA (arachidonoylethanolamide; also known as anandamide) participates in the neuroadaptations associated with chronic ethanol exposure. However, no studies have described the acute actions of ethanol on AEA production and degradation. In the present study, we investigated the time course of the effects of the intraperitoneal administration of ethanol (4 g/kg of body mass) on the endogenous levels of AEA in central and peripheral tissues. Acute ethanol administration decreased AEA in the cerebellum, the hippocampus and the nucleus accumbens of the ventral striatum, as well as in plasma and adipose tissue. Parallel decreases of a second acylethanolamide, PEA (palmitoylethanolamide), were observed in the brain. Effects were observed 45-90 min after ethanol administration. In vivo studies revealed that AEA decreases were associated with a remarkable inhibition of the release of both anandamide and glutamate in the nucleus accumbens. There were no changes in the expression and enzymatic activity of the main enzyme that degrades AEA, the fatty acid amidohydrolase. Acute ethanol administration did not change either the activity of N-acyltransferase, the enzyme that catalyses the synthesis of the AEA precursor, or the expression of NAPE-PLD (N-acylphosphatidylethanolamine-hydrolysing phospholipase D), the enzyme that releases AEA from membrane phospholipid precursors. These results suggest that receptor-mediated release of acylethanolamide is inhibited by the acute administration of ethanol, and that this effect is not derived from increased fatty acid ethanolamide degradation.


Comparison of Postoperative Hemorrhage Risk After Partial Liver Transplantation Versus Whole Liver Transplantation: A Single-Center Experience.
We aimed to identify risk factors associated with reoperation for postoperative intraperitoneal hemorrhage (PIH) after orthotopic liver transplantation and investigate if partial liver transplantation (PLT) increases the risk of PIH. We retrospectively analyzed the medical records of 304 consecutive recipients who underwent orthotopic liver transplantation at the Affiliated Lihuili Hospital, Ningbo University, from January 2016 to July 2022. Data were compared between recipients who experienced PIH requiring reoperation and those who did not. Subgroup propensity score matching analysis was performed to assess the impact of PLT on PIH risk. Neither prisoners nor participants who were coerced or paid were used in the study. Among the 304 recipients, 22 (7.2%) underwent reoperation for PIH. Multivariate analysis revealed that the recipient Model for End-Stage Liver Disease (MELD) score (odds ratio = 1.066, 95% CI [1.025-1.109], P = .001) and volume of intraoperative packed red blood cell transfusion (odds ratio = 1.089, 95% CI [1.032-1.481], P = .002) were independent risk factors for PIH. No significant differences were observed in the risk of PIH between PLT and whole liver transplantation. Preoperative MELD score and intraoperative packed red blood cell transfusion should be carefully considered to manage the risk of PIH in liver transplantation recipients. Partial liver transplantation, a crucial approach for addressing donor shortages, does not increase the risk of reoperation for PIH in recipients.


Mitophagy in Astrocytes Is Required for the Health of Optic Nerve.
Mitochondrial dysfunction in astrocytes has been implicated in the development of various neurological disorders. Mitophagy, mitochondrial autophagy, is required for proper mitochondrial function by preventing the accumulation of damaged mitochondria. The importance of mitophagy, specifically in the astrocytes of the optic nerve (ON), has been little studied. We introduce an animal model in which two separate mutations act synergistically to produce severe ON degeneration. The first mutation is in Cryba1, which encodes βA3/A1-crystallin, a lens protein also expressed in astrocytes, where it regulates lysosomal pH. The second mutation is in Bckdk, which encodes branched-chain ketoacid dehydrogenase kinase, which is ubiquitously expressed in the mitochondrial matrix and involved in the catabolism of the branched-chain amino acids. BCKDK is essential for mitochondrial function and the amelioration of oxidative stress. Neither of the mutations in isolation has a significant effect on the ON, but animals homozygous for both mutations (DM) exhibit very serious ON degeneration. ON astrocytes from these double-mutant (DM) animals have lysosomal defects, including impaired mitophagy, and dysfunctional mitochondria. Urolithin A can rescue the mitophagy impairment in DM astrocytes and reduce ON degeneration. These data demonstrate that efficient mitophagy in astrocytes is required for ON health and functional integrity.


The governance behaviours: a proposed approach for the alignment of the public and private sectors for better health outcomes.
Health systems are 'the ensemble of all public and private organisations, institutions and resources mandated to improve, maintain or restore health.' The private sector forms a major part of healthcare practice in many health systems providing a wide range of health goods and services, with significant growth across low-income and middle-income countries. WHO sees building stronger and more effective health systems through the participation and engagement of all health stakeholders as the pathway to further reducing the burden of disease and meeting health targets and the Sustainable Development Goals. However, there are governance and public policy gaps when it comes to interaction or engagement with the private sector, and therefore, some governments have lost contact with a major area of healthcare practice. As a result, market forces rather than public policy shape private sector activities with follow-on effects for system performance. While the problem is well described, proposed normative solutions are difficult to apply at country level to translate policy intentions into action. In 2020, WHO adopted a strategy report which argued for a major shift in approach to engage the private sector based on the performance of six governance behaviours. These are a practice-based approach to governance and draw on earlier work from Travis et al on health system stewardship subfunctions. This paper elaborates on the governance behaviours and explains their application as a practice approach for strengthening the capacity of governments to work with the private sector to achieve public policy goals.


Prospects of manipulating diatom silica nanostructure.
A key to the development of nanotechnology will be the ability to make complex nanoscaled three-dimensional structures at low cost and in large numbers. The wide variety of structures in the silicified cell walls of diatoms offers a promising natural source of such materials. Diatom silica can be converted into other materials, with maintenance of detailed morphology. To facilitate the use of diatoms in nanotechnology, specific manipulation of the structure in vivo will be desirable. This article explores the possibilities of manipulating diatom silica structure, by nongenetic and genetic means. Nongenetic influences that affect silica structure include changes in environmental conditions and life cycle stages and the presence or absence of particular compounds. The genetically based natural variation in structure in different diatom species indicates that genetic manipulation is possible. To achieve this, however, several goals must be met. The first is to identify cell wall synthesis (CWS) genes involved in structure formation. The recently completed genome sequence of Thalassiosira pseudonana opens the door for genomic and proteomic approaches to accomplish this. An important method to determine the function of CWS genes will be to modify gene sequences or expression and monitor the effect on structure. Performing gene modifications is straightforward, and modified genes can be introduced into diatoms, but the current inability to replace native diatom genes with modified copies could be a problem. However, there are feasible approaches yet to be applied to achieve this goal. It is very likely that continued development and application of molecular genetic techniques will enable us to specifically modify diatom silicified structures and provide a detailed understanding of the underlying mechanism of their formation.


Loss of Carnitine Palmitoyltransferase 1a Reduces Docosahexaenoic Acid-Containing Phospholipids and Drives Sexually Dimorphic Liver Disease in Mice.
Genome and epigenome wide association studies identified variants in carnitine palmitoyltransferase 1a (CPT1a) that associate with lipid traits. The goal of this study was to determine the impact by which liver-specific CPT1a deletion impacts hepatic lipid metabolism. Six-to-eight-week old male and female liver-specific knockout (LKO) and littermate controls were placed on a low-fat or high-fat diet (HFD; 60% kcal fat) for 15 weeks. Mice were necropsied after a 16 hour fast, and tissues were collected for lipidomics, matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI), kinome analysis, RNA-sequencing, and protein expression by immunoblotting. Female LKO mice had increased serum alanine aminotransferase (ALT) levels which were associated with greater deposition of hepatic lipids, while male mice were not affected by CPT1a deletion relative to male control mice. Mice with CPT1a deletion had reductions in DHA-containing phospholipids at the expense of monounsaturated fatty acids (MUFA)-containing phospholipids in both whole liver and at the level of the lipid droplet (LD). Male and female LKO mice increased RNA levels of genes involved in LD lipolysis ( Plin2 , Cidec , G0S2 ) and in polyunsaturated fatty acid (PUFA) metabolism ( Elovl5, Fads1, Elovl2 ), while only female LKO mice increased genes involved in inflammation ( Ly6d, Mmp12, Cxcl2 ). Kinase profiling showed decreased protein kinase A (PKA) activity, which coincided with increased PLIN2, PLIN5, and G0S2 protein levels and decreased triglyceride hydrolysis in LKO mice. Liver-specific deletion of CPT1a promotes sexually dimorphic steatotic liver disease (SLD) in mice, and here we have identified new mechanisms by which females are protected from HFD-induced liver injury.


Homing chemokines in rheumatoid arthritis.
In about 20% of patients with rheumatoid arthritis, B and T lymphocytes recruited into the inflamed synovium are organized into complex microstructures, which resemble secondary lymphoid organs. The development of such lymphoid aggregates with germinal centers appears to contribute to the pathogenesis of the disease. Growing evidence indicates that chemokines and their receptors control the recruitment and positioning of leukocytes as well as their organization into node-like lymphoid structures. Here, we comment on recent studies highlighting the importance of chemokines in rheumatoid arthritis, in particular of B-cell-activating chemokine-1 in lymphoid neogenesis in the inflamed synovium.


Increased expression of TRPC4 channels associated with erectile dysfunction in diabetes.
In recent reports, an association between altered TRPC channel function and the development of various diabetic complications has drawn the attention of many investigators. The aim of this study was to investigate the expression of TRPC4 channels of corpus smooth muscle (CSM) cells in diabetes, and to evaluate the association between erectile dysfunction (ED) and altered TRPC4 channel function. The expression of TRPC4 in the penile tissue of human, normal and diabetic rat was investigated using RT-PCR, western blotting and immunohistochemistry (IHC). In vivo gene transfer of dominant negative (DN) TRPC4 into the CSM of rat was conducted. In vivo pelvic nerve stimulation was performed to measure erectile function. Expression of TRPC1, TRPC3, TRPC4 and TRPC6 in human and rat CSM tissues was confirmed by RT-PCR, western blot and IHC. In the diabetic rat, the expression levels of mRNA and protein of the TRPC4, and TRPC6 were significantly increased compared to control rats (p < 0.05). The change in TRPC4 expression in the diabetic rats was higher than those of the other TRPC subunits (p < 0.05). The IHC showed that only TRPC4 expression had a higher intensity in the diabetes compared to normal rats (p < 0.05). Gene transfection with TRPC4(DN) into the diabetic rats restored erectile function to levels similar to that of normal controls. Gene expression of TRPC4(DN) in CSM tissue was confirmed by RT-PCR 2 weeks after transfection. This study demonstrated that TRPC4 channel expression increased in the penile CSM cells of diabetic rats. The down-regulation of TRPC4 with DN form restored erectile function in the diabetic rats. The alteration of TRPC4 channel is one of pathophysiology of ED and could be a target for drug development for ED.


Should we use a case-crossover design?
The first decade of experience with case-crossover studies has shown that the design applies best if the exposure is intermittent, the effect on risk is immediate and transient, and the outcome is abrupt. However, this design has been used to study single changes in exposure level, gradual effects on risk, and outcomes with insidious onsets. To estimate relative risk, the exposure frequency during a window just before outcome onset is compared with exposure frequencies during control times rather than in control persons. One or more control times are supplied by each of the cases themselves, to control for confounding by constant characteristics and self-confounding between the trigger's acute and chronic effects. This review of published case-crossover studies is designed to help the reader prepare a better research proposal by understanding triggers and deterrents, target person times, alternative study bases, crossover cohorts, induction times, effect and hazard periods, exposure windows, the exposure opportunity fallacy, a general likelihood formula, and control crossover analysis.


Non-treatment-related chronic myeloid leukemia as a second malignancy.
The characteristics of the very rare non-treatment-related chronic myeloid leukemia (nTr-CML) cases have never before been analyzed. The literature up to December 2002 was screened using the Medline database to identify cases of Tr-CML and nTr-CML. We considered five cases with nTr-CML identified among 270 newly diagnosed CML at our Department. Our report thus considers nine cases with nTr-CML compared to 77 affected by Tr-CML as a secondary neoplasm. The median age at the appearance of the first tumor was higher in nTr-CML patients compared to that of the Tr-CML group (P<0.0001). The median age at CML diagnosis was significantly higher in the nTr-CML than in the Tr-CML group (P<0.0001). The proportion of hematological malignancies as first tumor type was not different in the two groups (44% in nTr-CML versus 56% in Tr-CML). Our study underlines that nTr-CML as a second malignancy is a rare entity associated with elderly age.


Effect of signal-temporal uncertainty in children and adults: tone detection in noise or a random-frequency masker.
A cue indicating when in time to listen can improve adults' tone detection thresholds, particularly for conditions that produce substantial informational masking. The purpose of this study was to determine if 5- to 13-yr-old children likewise benefit from a light cue indicating when in time to listen for a masked pure-tone signal. Each listener was tested in one of two continuous maskers: Broadband noise (low informational masking) or a random-frequency, two-tone masker (high informational masking). Using a single-interval method of constant stimuli, detection thresholds were measured for two temporal conditions: (1) Temporally-defined, with the listening interval defined by a light cue, and (2) temporally-uncertain, with no light cue. Thresholds estimated from psychometric functions fitted to the data indicated that children and adults benefited to the same degree from the visual cue. Across listeners, the average benefit of a defined listening interval was 1.8 dB in the broadband noise and 8.6 dB in the random-frequency, two-tone masker. Thus, the benefit of knowing when in time to listen was more robust for conditions believed to be dominated by informational masking. An unexpected finding of this study was that children's thresholds were comparable to adults' in the random-frequency, two-tone masker.


Exploring the strategies for successfully building e-portfolios in medical schools.
The purpose of this study is to examine the characteristics of resistance among medical students toward e-portfolios and find the strategies for them to successfully prepare e-portfolios. Participants were a group of 258 medical students. The questionnaire comprised 13 items developed based on the innovation resistance theory. The data were analyzed using descriptive analysis and Spearman's correlation analysis using PASW SPSS version 18.0 (SPSS Inc., Chicago, USA). Students perceived that e-portfolios have a high degree of relative advantage, trialability, and complexity as their innovation resistance characteristics. Regarding perceived risk, they did not want others to see their information, but they had a high degree of demand for communication with their professors. The successful use of portfolios can serve as a tool for student management and assessment that can reflect their introspection, personal development, and academic performance. This study proposed the strategies to promote the use of e-portfolios by strengthening education on the need for them, specific feedback of instructors, and students' autonomy.


The Association of Laboratory Test Abnormalities With Mortality Risk in Pediatric Intensive Care.
To determine the bivariable associations between abnormalities of 28 common laboratory tests and hospital mortality and determine how mortality risks changes when the ranges are evaluated in the context of commonly used laboratory test panels. A 2009-2016 cohort from the Health Facts (Cerner Corporation, Kansas City, MO) database. Hospitals caring for children in ICUs. Children cared for in ICUs with laboratory data. None. There were 2,987,515 laboratory measurements in 71,563 children. The distribution of laboratory test values in 10 groups defined by population percentiles demonstrated the midrange of tests was within the normal range except for those measured predominantly when significant abnormalities are suspected. Logistic regression analysis at the patient level combined the population-based groups into ranges with nonoverlapping mortality odds ratios. The most deviant test ranges associated with increased mortality risk (mortality odds ratios > 5.0) included variables associated with acidosis, coagulation abnormalities and blood loss, immune function, liver function, nutritional status, and the basic metabolic profile. The test ranges most associated with survival included normal values for chloride, pH, and bicarbonate/total Co2. When the significant test ranges from bivariable analyses were combined in commonly used test panels, they generally remained significant but were reduced as risk was distributed among the tests. The relative importance of laboratory test ranges vary widely, with some ranges strongly associated with mortality and others strongly associated with survival. When evaluated in the context of test panels rather than isolated tests, the mortality odds ratios for the test ranges decreased but generally remained significant as risk was distributed among the components of the test panels. These data are useful to develop critical values for children in ICUs, to identify risk factors previously underappreciated, for education and training, and for future risk score development.


Glyphosate-Resistant and Conventional Canola (Brassica napus L.) Responses to Glyphosate and Aminomethylphosphonic Acid (AMPA) Treatment.
Glyphosate-resistant (GR) canola contains two transgenes that impart resistance to the herbicide glyphosate: (1) the microbial glyphosate oxidase gene (gox) encoding the glyphosate oxidase enzyme (GOX) that metabolizes glyphosate to aminomethylphosphonic acid (AMPA) and (2) cp4 that encodes a GR form of the glyphosate target enzyme 5-enolpyruvylshikimic acid-3-phosphate synthase. The objectives of this research were to determine the phytotoxicity of AMPA to canola, the relative metabolism of glyphosate to AMPA in GR and conventional non-GR (NGR) canola, and AMPA pool sizes in glyphosate-treated GR canola. AMPA applied at 1.0 kg ha(-1) was not phytotoxic to GR or NGR. At this AMPA application rate, NGR canola accumulated a higher concentration of AMPA in its tissues than GR canola. At rates of 1 and 3.33 kg ae ha(-1) of glyphosate, GR canola growth was stimulated. This stimulatory effect is similar to that of much lower doses of glyphosate on NGR canola. Both shikimate and AMPA accumulated in tissues of these glyphosate-treated plants. In a separate experiment in which young GR and NGR canola plants were treated with non-phytotoxic levels of [(14)C]-glyphosate, very little glyphosate was metabolized in NGR plants, whereas most of the glyphosate was metabolized to AMPA in GR plants at 7 days after application. Untreated leaves of GR plants accumulated only metabolites (mostly AMPA) of glyphosate, indicating that GOX activity is very high in the youngest leaves. These data indicate that more glyphosate is transformed to AMPA rapidly in GR canola and that the accumulated AMPA is not toxic to the canola plant.


An efficient method for removing bilirubin.
Bilirubin is tightly bound to albumin, making hemoperfusion an ineffective treatment for hyperbilirubinemia. By adding a safe unbinding agent to the blood (solutizer), which itself is adsorbed, hemoperfusion can become efficient and practical. Canines were made hyperbilirubinemic with an intravenous infusion of a 5 mg/ml solution (with Na2CO3) for 1 hour. Peak concentrations of 14-22 mg/dl were reached in adult dogs (25-35 kg). Hemoperfusion was then initiated with or without (control) the solutizer (sodium benzoate). The bilirubin unbinding effect of sodium benzoate was rapid and effective. Because of the simultaneous adsorption of sodium benzoate, a small activated carbon section that was presaturated with the solutizer was located proximal to the main hemoperfusion column, in addition to continuous infusion to reach 20 mM in the blood. Comparison of the normalized bilirubin concentration for benzoate augmented hemoperfusion with the average for control dogs, shows that benzoate results in a threefold decrease in the normalized bilirubin concentration after 1.5 hr of hemoperfusion. Sodium benzoate may also have the advantage of protecting platelets during hemoperfusion.


Physical activity and exercise-induced bronchoconstriction in Greek schoolchildren.
To investigate the association between physical activity and exercise-induced bronchoconstriction (EIB) in an urban population sample of schoolchildren, taking into account potential confounders such as asthma symptoms and overweight. Children aged 10-12 years answered validated questionnaires on physical activity (Physical Activity and Lifestyle Questionnaire) and asthma symptoms (ISAAC questionnaire), and were categorized according to their body mass index (BMI). EIB (FEV(1) decrease from baseline ≥13%) was assessed by a standardized free running Exercise Challenge Test (ECT). Six hundred seven children completed the ECT. There were no differences among asthma groups (diagnosed asthma, asthma-related symptoms not diagnosed as asthma, no asthma-related symptoms) regarding total daily energy expenditure and time spent in mild (1.1-2.9 metabolic equivalents-METs), moderate (3-6 METs), and vigorous (>6 METs) activities. Only overweight/obese EIB-positive children had shorter duration of vigorous activity as compared to their EIB-negative or non-overweight/obese EIB-positive peers. Total daily energy expenditure and duration of mild- and moderate-intensity activity were negatively associated with EIB independently of BMI status or asthma-related symptoms. Decreased levels of physical activity are associated with EIB irrespectively of BMI status and asthma-related symptoms. Longitudinal studies are needed to confirm the negative impact of sedentary lifestyle on the development of EIB suggested by these findings.


Ice slurry ingestion increases core temperature capacity and running time in the heat.
To investigate the effect of ice slurry ingestion on thermoregulatory responses and submaximal running time in the heat. On two separate occasions, in a counterbalanced order, 10 males ingested 7.5 g·kg(-1) of either ice slurry (-1°C) or cold water (4°C) before running to exhaustion at their first ventilatory threshold in a hot environment (34.0°C ± 0.2°C, 54.9% ± 5.9% relative humidity). Rectal and skin temperatures, HR, sweating rate, and ratings of thermal sensation and perceived exertion were measured. Running time was longer (P = 0.001) after ice slurry (50.2 ± 8.5 min) versus cold water (40.7 ± 7.2 min) ingestion. Before running, rectal temperature dropped 0.66°C ± 0.14°C after ice slurry ingestion compared with 0.25°C ± 0.09°C (P = 0.001) with cold water and remained lower for the first 30 min of exercise. At exhaustion, however, rectal temperature was higher (P = 0.001) with ice slurry (39.36°C ± 0.41°C) versus cold water ingestion (39.05°C ± 0.37°C). During exercise, mean skin temperature was similar between conditions (P = 0.992), as was HR (P = 0.122) and sweat rate (P = 0.242). After ice slurry ingestion, subjects stored more heat during exercise (100.10 ± 25.00 vs 78.93 ± 20.52 W·m(-2), P = 0.005), and mean ratings of thermal sensation (P = 0.001) and perceived exertion (P = 0.022) were lower. Compared with cold water, ice slurry ingestion lowered preexercise rectal temperature, increased submaximal endurance running time in the heat (+19% ± 6%), and allowed rectal temperature to become higher at exhaustion. As such, ice slurry ingestion may be an effective and practical precooling maneuver for athletes competing in hot environments.


Extra centrosomes and/or chromosomes prolong mitosis in human cells.
Using laser microsurgery and cell fusion we have explored how additional centrosomes and/or chromosomes influence the duration of mitosis in human cells. We found that doubling the chromosome number added approximately 10 min to a 20 min division, whereas doubling the number of centrosomes added approximately 30 min more. Extra centrosomes and/or chromosomes prolong mitosis by delaying satisfaction of the spindle assembly checkpoint. Thus mitosis can be prolonged by non-genetic means and extra chromosomes and centrosomes probably contribute to the elevated mitotic index seen in many tumours.


Sequence and expression of GLN3, a positive nitrogen regulatory gene of Saccharomyces cerevisiae encoding a protein with a putative zinc finger DNA-binding domain.
The GLN3 gene of Saccharomyces cerevisiae is required for the activation of transcription of a number of genes in response to the replacement of glutamine by glutamate as source of nitrogen. We cloned the GLN3 gene and constructed null alleles by gene disruption. GLN3 is not essential for growth, but increased copies of GLN3 lead to a drastic decrease in growth rate. The complete nucleotide sequence of the GLN3 gene was determined, revealing one open reading frame encoding a polypeptide of 730 amino acids, with a molecular weight of approximately 80,000. The GLN3 protein contains a single putative Cys2/Cys2 zinc finger which has homology to the Neurospora crassa NIT2 protein, the Aspergillus nidulans AREA protein, and the erythroid-specific transcription factor GATA-1. Immunoprecipitation experiments indicated that the GLN3 protein binds the nitrogen upstream activation sequence of GLN1, the gene encoding glutamine synthetase. Neither control of transcription nor control of initiation of translation of GLN3 is important for regulation in response to glutamine availability.


Expression of calcium/calmodulin-dependent protein kinase in relation to dopamine islands and synaptic maturation in the cat striatum.
During pre- and postnatal development the dopamine-containing nigrostriatal afferents of the striatum are arranged as a conspicuous series of patches (the "dopamine islands"). The development of this dopamine island system, which metamorphoses in early postnatal life to the striosomal architecture of the adult, has recently received considerable attention, but the factors initiating and influencing maturation of this architecture are largely unknown. In an attempt to clarify the relationships between the onset of clustering of dopamine-containing afferents, the grouping of neurons within future striosomes and the maturation of synapses in the striatum, we compared the initial prenatal appearance and subsequent development of immunohistochemical markers for the dopamine-containing innervation [tyrosine hydroxylase (TH)-like immunoreactivity], for synaptic vesicles (SV48-like immunoreactivity), and for a phosphorylation-related enzyme Ca2+/calmodulin-dependent protein kinase type II (CaM kinase II-like immunoreactivity) that is expressed in virtually all striatal neurons by adulthood. Here we present evidence that during striatal ontogeny, both neurons and neuropil expressing CaM kinase II-like immunoreactivity and SV48-positive terminals form discrete patches that are in register with dopamine islands. It is CaM kinase II-positive elements, however, rather than the TH-positive island fibers (or SV48-positive synapses), that initially form overt clusters. Dopamine-containing fibers begin to innervate the striatal anlage just prior to embryonic day (E) 32. Their distribution follows the general lateral to medial developmental gradient characteristic of the striatum but is not yet distinctly islandic. At this time, CaM kinase II-like immunoreactivity was very weak or not present at all and SV48-like immunoreactivity was undetectable. By E36, CaM kinase II-positive neurons are visible in discrete patches of immunopositive neuropil, but only faint inhomogeneities are detectable in the distribution of TH-positive fibers and scarcely any SV48-like immunoreactivity can be seen. By E45, all 3 markers are focused in typical islandic patterns, and they remain so into the early postnatal period. These observations suggest a developmental sequence in which dopamine-containing fibers invade the striatal anlage prior to forming distinct islandic foci and prior to the maturational events signaled by the production of CaM kinase II within the neurons and neuropil of future striosomes.(ABSTRACT TRUNCATED AT 400 WORDS)


Structure, function, and tethering of DNA-binding domains in σ⁵⁴ transcriptional activators.
We compare the structure, activity, and linkage of DNA-binding domains (DBDs) from σ(54) transcriptional activators and discuss how the properties of the DBDs and the linker to the neighboring domain are affected by the overall properties and requirements of the full proteins. These transcriptional activators bind upstream of specific promoters that utilize σ(54)-polymerase. Upon receiving a signal the activators assemble into hexamers, which then, through adenosine triphosphate (ATP) hydrolysis, drive a conformational change in polymerase that enables transcription initiation. We present structures of the DBDs of activators nitrogen regulatory protein C 1 (NtrC1) and Nif-like homolog 2 (Nlh2) from the thermophile Aquifex aeolicus. The structures of these domains and their relationship to other parts of the activators are discussed. These structures are compared with previously determined structures of the DBDs of NtrC4, NtrC, ZraR, and factor for inversion stimulation. The N-terminal linkers that connect the DBDs to the central domains in NtrC1 and Nlh2 were studied and found to be unstructured. Additionally, a crystal structure of full-length NtrC1 was solved, but density of the DBDs was extremely weak, further indicating that the linker between ATPase and DBDs functions as a flexible tether. Flexible linking of ATPase and DBDs is likely necessary to allow assembly of the active hexameric ATPase ring. The comparison of this set of activators also shows clearly that strong dimerization of the DBD only occurs when other domains do not dimerize strongly.


Association between host species choice and morphological characters of main sensory structures of Culicoides in the Palaeartic region.
Culicoides (Diptera: Ceratopogonidae) serve as vectors of several mammalian and avian diseases, including bluetongue, Schmallenberg, African horse sickness, avian malaria and Oropouche. Host preference investigations are necessary to assess the transmission routes of vector-borne diseases and to inform mitigation strategies. A recent study examining the main sensory structures (palps and antennae) of Culicoides species suggests that they be classified as ornithophilic or mammalophilic according to their feeding habits. We analyzed Culicoides host preferences according to the literature and carried out a multiple correspondence analysis linking these preferences with morphological data. Seven out of 12 variables were found to be reliable predictors of host preference in Culicoides species: Antenna Flagellomer-Sensilla Coeloconica-Number: (7-10) and (11-13); Antenna Flagellomer-Sensilla Coeloconica IV-X: presence; Palpus-size: wide and/or narrow opening and shallow pit; Palpus-Shape: strongly swollen; Antenna-Short sensilla trichodea-distal part segment IV to X-Number: 2 seta each. Our results demonstrate that the presence of sensilla coeloconica and the maxillary palpus can be used to separate ornithophilic and mammalophilic or ornithophilic/mammalophilic species.