Maximal oxygen consumption is best predicted by measures of cardiac size rather than function in healthy adults.
Training induces changes in cardiac structure and function which improves cardiac output (CO) and oxygen delivery during exercise. It is unclear whether it is cardiac structure or function which is of greatest importance in determining maximal oxygen consumption (VO(2max)). In 55 subjects (15 non-athletes, 32 amateur and 8 elite athletes), left and right ventricular (LV and RV) volumes and mass were assessed by magnetic resonance imaging (CMR). Comprehensive traditional and novel echocardiographic measures included colour-coded Doppler echocardiography to assess myocardial velocities, strain and strain rate at rest and maximal exercise in both ventricles. Measures of cardiac size and function were assessed as univariate and multivariate predictors of VO(2max). LV and RV mass correlated strongly with VO(2max) (r = 0.79 and r = 0.65, respectively, p < 0.0001), as did LV and RV end-diastolic volumes (r = 0.68 and r = 0.75, p < 0.0001) and heart rate reserve (r = 0.60, p < 0.0001). Measures of myocardial function were not predictive of VO(2max) with the exception of RV diastolic velocities (r = 0.32 and r = 0.36 for rest and exercise, respectively, p < 0.05). On multivariate analysis, only RV end-diastolic volume, LV mass and heart rate reserve were independent predictors (beta = 0.28, 0.45 and 0.27 respectively, p < 0.0001) and together explained 73% of the variance in VO(2max). Measures of cardiac morphology are strongly associated with VO(2max) in healthy adults and well-trained athletes. A combination of ventricular volume, mass and heart rate reserve explains much of the variance in VO(2max), whilst measures of myocardial function do not further strengthen predictive models.


Differences in serum LH and FSH levels using depot or daily GnRH agonists in controlled ovarian stimulation: influence on ovarian response and outcome of ART.
To study effects of endogenous LH levels on ovarian response and outcome in ART cycles a controlled study was performed with two patient groups differing in the intensity of pituitary downregulation. Group I (n = 27) received 3.75 mg of the GnRH agonist triptorelin acetate depot, group II (n = 54) was given 0.1 mg triptorelin acetate daily, followed by ovarian stimulation with recombinant FSH. After downregulation serum LH and FSH levels were significantly lower in group I. Patients of group I needed significantly higher FSH doses to achieve comparable levels of serum estradiol and preovulatory follicles. The number of retrieved oocytes and transferable embryos was lower in group I. Patients with profound endogenous LH suppression by depot GnRH agonists show higher FSH stimulation dose requirements and lower oocyte number and fertilization rate, indicating a need for minimal LH activity in folliculogenesis and oocyte development.


Animal model for angiotensin II effects in the internal anal sphincter smooth muscle: mechanism of action.
Effect of ANG II was investigated in in vitro smooth muscle strips and in isolated smooth muscle cells (SMC). Among different species, rat internal and sphincter (IAS) smooth muscle showed significant and reproducible contraction that remained unmodified by different neurohumoral inhibitors. The AT(1) antagonist losartan but not AT(2) antagonist PD-123319 antagonized ANG II-induced contraction of the IAS smooth muscle and SMC. ANG II-induced contraction of rat IAS smooth muscle and SMC was attenuated by tyrosine kinase inhibitors genistein and tyrphostin, protein kinase C (PKC) inhibitor H-7, Ca(2+) channel blocker nicardipine, Rho kinase inhibitor Y-27632 or p(44/42) mitogen-activating protein kinase (MAPK(44/42)) inhibitor PD-98059. Combinations of nicardipine and H-7, Y-27632, and PD-98059 caused further attenuation of the ANG II effects. Western blot analyses revealed the presence of both AT(1) and AT(2) receptors. We conclude that ANG II causes contraction of rat IAS smooth muscle by the activation of AT(1) receptors at the SMC and involves multiple intracellular pathways, influx of Ca(2+), and activation of PKC, Rho kinase, and MAPK(44/42).


Nitrogen-doped carbon dots: Recent developments in its fluorescent sensor applications.
Doped carbon dots have attracted great attention from researchers across disciplines because of their unique characteristics, such as their low toxicity, physiochemical stability, photostability, and outstanding biocompatibility. Nitrogen is one of the most commonly used elements for doping because of its sizeable atomic radius, strong electronegativity, abundance, and availability of electrons. This distinguishes them from other atoms and allows them to perform distinctive roles in various applications. Here, we have reviewed the most current breakthroughs in nitrogen-doped CDs (N-CDs) for fluorescent sensor applications in the last five years. The first section of the article addresses several synthetic and sustainable ways of making N-CDs. Next, we briefly reviewed the fluorescent features of N-CDs and their sensing mechanism. Furthermore, we have thoroughly reviewed their fluorescent sensor applications as sensors for cations, anions, small molecules, enzymes, antibiotics, pathogens, explosives, and pesticides. Finally, we have discussed the N-CDs' potential future as primary research and how that may be used. We hope that this study will contribute to a better understanding of the principles of N-CDs and the sensory applications that they can serve.


Predicting three-dimensional structure of protein fragments from dihedral angle propensities and molecular dynamics.
Incorporating the existing knowledge of protein structural preferences, e.g., amino acid angle frequencies, in structure prediction have proven to be less successful with smaller peptides. In this work, we compare the effectiveness of backbone angle propensity libraries derived from two protein data sets: one consisting of proteins of unrestricted lengths; the second containing proteins ranging in size from 40 to 75 residues. Model structures for 29 target peptides are predicted using a threading algorithm and their stability evaluated using in vacuo molecular dynamics simulations. Structures derived from the data set consisting of smaller proteins outperformed those developed from that unrestricted by protein length.


Changes in the acoustic environment alter the foraging and sheltering behaviour of the cichlid Amititlania nigrofasciata.
Anthropogenic noise can affect behaviour across a wide range of species in both terrestrial and aquatic environments. However, behaviours might not be affected in isolation. Therefore, a more holistic approach investigating how environmental stressors, such as noise pollution, affect different behaviours in concert is necessary. Using tank-based noise exposure experiments, we tested how changes in the acoustic environment affect the behaviour of the cichlid Amatitlania nigrofasciata. We found that exposure to anthropogenic noise affected a couple of behaviours: an increase in sheltering was accompanied by a decrease in foraging. Our results highlight the multiple negative effects of an environmental stressor on an individual's behaviour.


Extracellular synthesis of silver nanoparticles by the Bacillus strain CS 11 isolated from industrialized area.
Biological synthesis of silver nanoparticles using microorganisms has received profound interest because of their potential to synthesize nanoparticles of various size, shape and morphology. In the current study, synthesis of silver nanoparticles by a bacterial strain (CS 11) isolated from heavy metal contaminated soil is reported. Molecular identification of the isolate showed it as a strain of Bacillus sp. On treating the bacteria with 1 mM AgNO3, it was found to have the ability to form silver nanoparticles extracellularly at room temperature within 24 h. This was confirmed by the visual observation and UV-Vis absorption at 450 nm. Further characterization of nanoparticles by transmission electron microscopy confirmed the size of silver nanoparticles in 42-92 nm range. Therefore, the current study is a demonstration of an efficient synthesis of stable silver nanoparticle by a Bacillus strain.


Accuracy and reproducibility of preoperative three-dimensional planning for total hip arthroplasty using biplanar low-dose radiographs : A pilot study.
In total hip arthroplasty (THA), the acetabular cup and femoral stem must be correctly sized and positioned to avoid intraoperative and postoperative complications, achieve good functional outcomes and ensure long-term survival. Current two-dimensional (2D) techniques do not provide sufficient accuracy, while low-dose biplanar X-rays (EOS) had not been assessed in this indication. Therefore, we performed a case-control study to : (1) evaluate the prediction of stem and cup size for a new 3D planning technique (stereoradiographic imaging plus 3D modeling) in comparison to 2D templating on film radiographs and (2) evaluate the accuracy and reproducibility of this 3D technique for preoperative THA planning. Accuracy and reproducibility are better with the 3D vs. 2D method. Stem and cup sizes were retrospectively determined by two senior surgeons, twice, for a total of 31 unilateral primary THA patients in this pilot study, using 3D preplanning software on low-dose biplanar X-rays and with 2D templating on conventional anteroposterior (AP) film radiographs. Patients with a modular neck or dual-mobility prosthesis were excluded. All patients but one had primary osteoarthritis; one following trauma did not have a cup implanted. The retrospectively planned sizes were compared to the sizes selected during surgery, and intraclass coefficients (ICC) calculated. 3D planning predicted stem size more accurately than 2D templating: stem sizes were planned within one size in 26/31 (84%) of cases in 3D versus 21/31 (68%) in 2D (P=0.04). 3D and 2D planning accuracies were not significantly different for cup size: cup sizes were planned within one size in 28/30 (92%) of cases in 3D versus 26/30 (87%) in 2D (P=0.30). ICC for stem size were 0.88 vs. 0.91 for 3D and 2D, respectively. Inter-operator ICCs for cup size were 0.84 vs. 0.71, respectively. Repetitions of the 3D planning were within one size (except one stem), with the majority predicting the same size. Increased accuracy in 3D may be due to the use of actual size (non-magnified) images, and judging fit on AP and lateral images simultaneously. Results for other implant components may differ from those presented. Size selection may improve further with planning experience, based on a feedback loop between planning and surgical execution. Level III. Retrospective case-control study.


Eicosapentaenoic acid supplementation modulates the osteoblast/osteoclast balance in inflammatory environments and protects against estrogen deficiency-induced bone loss in mice.
An imbalance of osteoblasts (OBs) and osteoclasts (OCs) in a chronic inflammatory microenvironment is an important pathological factor leading to osteoporosis. Eicosapentaenoic acid (EPA) has been shown to suppress inflammation in macrophages and adipocytes. However, the effect of EPA on OBs and OCs has yet to be fully elucidated. We explored the roles of EPA in the differentiation of OBs and OCs, as well as the coupling between OBs and OCs in an inflammatory microenvironment. The effects of EPA on estrogen deficiency-induced osteoporosis were also evaluated. Mouse bone marrow mesenchymal stem cells (mBMSCs) and mouse bone marrow-derived macrophages (mBMMs) were used for in vitro OBs and OCs differentiation. TNF-α was used to create an inflammatory microenvironment. We examined the effects of EPA on osteoblastogenesis in the absence or presence of TNF-α and collect OBs' culture medium as the conditioned medium (CM). Then we examined the effects of EPA and CM on RANKL-induced osteoclastogenesis. The in vivo effects of EPA were determined using an ovariectomized (OVX) mouse model treated with EPA or vehicle. High-dose EPA was shown to promote osteoblastogenesis in an inflammatory environment in vitro, as well as upregulate expression of OBs-specific proteins and genes. ARS and ALP staining also showed that high-dose EPA-treated groups restored mBMSCs' impaired osteogenic capacity caused by TNFa. Mechanistically, EPA suppressed the NF-κB pathway activated by TNF-α in mBMSCs and rescued TNF-α-mediated inhibition of osteoblastogenesis. EPA was also shown to inhibit expression of RANKL and decrease the RANKL/OPG ratio in OBs in an inflammatory environment. CM from TNF-α-stimulated OBs promoted osteoclastogenesis of mBMMs; EPA-treated CM prevented this. In the OVX mouse model, EPA supplementation prevented bone loss in an estrogen deficiency-induced inflammatory environment. EPA was demonstrated for the first time to restore mBMSCs' impaired osteogenic capacity caused by TNFa-induced inflammation and rescue the OBs/OCs balance via regulation of RANKL and OPG expression in OBs. EPA showed a remarkable ability to prevent bone loss in OVX mice, suggesting a potential application of EPA in postmenopausal osteoporosis.


Impact of remission induction chemotherapy on survival in older adults with acute myeloid leukemia.
Significant controversy surrounds the use of remission induction chemotherapy (IC) in older adults with acute myeloid leukemia (AML). Earlier clinical trials have yielded conflicting results and possibly a minor survival benefit, often offset by a longer hospitalization time. To evaluate the role of IC in patients with AML, a case control study of patients 60 years or older treated at the Cleveland Clinic Taussig Cancer Center between 1997 and 2005 was conducted. Forty-four patients who did not receive IC were matched by a propensity analysis to 138 patients who received an anthracycline-based regimen. The unadjusted median survival of patients who did not receive IC was 53 days, compared with 197 days (P < .001) for those who did. After further adjusting for age, gender, race, leukocyte count at presentation, AML cytogenetics, history of prior hematologic disorder, and assessing for comorbidities, not receiving IC was still associated with worse survival (hazards ratio of 1.88; 95% confidence interval, 1.15-3.05 [P = .01]). Additional predictors of poor outcomes in older adults with AML included higher leukocyte count at presentation, poor-risk cytogenetics, and African-American race (compared with Caucasians). The study suggests improved outcomes in older adults with AML who undergo remission induction therapy.


The palaeoecology of the micromammals from the late middle Pleistocene site of Hoedjiespunt 1 (Cape Province, South Africa).
The palaeontological site of Hoedjiespunt 1 (HDP1) represents a fossilized hyaena lair. A rich mammalian fauna, including four hominid teeth, has been recovered from the site. Micromammals were recovered from the same sediments as the larger fauna. Taphonomic analysis suggests that the micromammal assemblages from HDP1 were accumulated by a barn owl. The barn owl produces micromammal assemblages that provide a broad sample of micromammals, within a certain size range, living in the hunting area of the owl. There are size-related and other biases inherent in the prey selection of this predator, and owls may roost in one area and hunt in another however, the barn owl has frequently been found to provide a better indication of micromammals living within an area than trapping. The micromammals from HDP1 were used to reconstruct the microhabitats in the vicinity of the site. Two taxonomic habitat indexes were used to assess the environment and dominant habitat types at Hoedjiespunt 1. The variability and adaptability of many of the southern African micromammals complicates interpretation of the results, however, it appears that the micromammals from the HDP1 fossil assemblages utilized habitats of open, scrub vegetation, and rocky and sandy areas. It is suggested that the environment was not markedly different from today, but it may have been relatively more arid. A comparison between HDP1 and other fossil sites in the area dating from the terminal Pleistocene to the Holocene indicates that HDP1 is lacking certain species that are common to all the other west coast fossil sites. There is some discrepancy in the environment indicated by the large mammals as compared that indicated by to the micromammals at the site. It is suggested that this discrepancy may reflect the fact that an owl is likely to have hunted in the vicinity of the hyaena den, probably in the more open areas around the roost site, whereas the macrofauna, accumulated by the further-ranging brown hyaena (Hyaena brunnea), represents environments from further afield.


Are there causal relationships between attention-deficit/hyperactivity disorder and body mass index? Evidence from multiple genetically informed designs.
Attention-deficit/hyperactivity disorder (ADHD) and body mass index (BMI) are associated. However, it remains unclear whether this association reflects causal relationships in either direction or confounding. Here, we implemented genetically informed methods to examine bidirectional causality and potential confounding. Three genetically informed methods were employed: (i) cross-lagged twin-differences analyses to assess bidirectional effects of ADHD symptoms and BMI at ages 8, 12, 14 and 16 years in 2386 pairs of monozygotic twins from the Twins Early Development Study (TEDS); (ii) within- and between-family ADHD and BMI polygenic score (PS) analyses in 3320 pairs of dizygotic TEDS twins; and (iii) two-sample bidirectional Mendelian randomization (MR) using summary statistics from genome-wide association studies (GWAS) on ADHD (N = 55,374) and BMI (N = 806,834). Mixed results were obtained across the three methods. Twin-difference analyses provided little support for cross-lagged associations between ADHD symptoms and BMI over time. PS analyses were consistent with bidirectional relationships between ADHD and BMI, with plausible time-varying effects from childhood to adolescence. MR findings also suggested bidirectional causal effects between ADHD and BMI. Multivariable MR indicated the presence of substantial confounding in bidirectional relationships. The three methods converged to highlight multiple sources of confounding in the association between ADHD and BMI. PS and MR analyses suggested plausible causal relationships in both directions. Possible explanations for mixed causal findings across methods are discussed.


Plants and water in a changing world: a physiological and ecological perspective.
The reduction of greenhouse gases (GHGs) emission by replacing fossil energy stocks with carbon-neutral fuels is a major topic of the political and scientific debate on environmental sustainability. Such shift in energy sources is expected to curtail the accumulation rate of atmospheric CO2, which is a strong infrared absorber and thus contributes to the global warming effect. Although such change would produce desirable outputs, the consequences of a drastic decrease in atmospheric CO2 (the substrate of photosynthesis) should be carefully considered in the light of its potential impact on ecosystems stability and agricultural productivity. Indeed, plants regulate CO2 uptake and water loss through the same anatomical structure: the leaf stomata. A reduced CO2 availability is thus expected to enhance transpiration rate in plants decreasing their water use efficiency and imposing an increased water demand for both agricultural and wild ecosystems. We suggest that this largely underestimated issue should be duly considered when implementing policies that aim at the mitigation of global environmental changes and, at the same time, promote sustainable agricultural practices, include the preservation of biodiversity. Also, we underlie the important role(s) that modern biotechnology could play to tackle these global challenges by introducing new traits aimed at creating crop varieties with enhanced CO2 capture and water- and light-use efficiency.


Distinct detergent-resistant membrane microdomains (lipid rafts) respectively harvest K(+) and water transport systems in brain astroglia.
The detergent-resistant microdomains (DRM) of cell membranes scaffold different molecules and regulate cell functions by orchestrating various signaling pathways including G-proteins and tyrosine kinase. Here we report a novel role for DRM in astroglial cells. K(+)-buffering inwardly rectifying Kir4.1 channels and the water channel AQP4 are expressed in astrocytes and they may be functionally coupled to maintain ionic and osmotic homeostasis in the brain. Kir4.1 and AQP4 channel proteins were abundant in noncaveloar DRM in the brain and also in HEK293T cells when exogenously expressed. In HEK293T cells, depletion of membrane cholesterol by methyl-beta-cyclodextrin (MbetaCD) resulted in loss of Kir4.1 association with DRM and its channel activity but did not affect either the distribution or the function of AQP4. Immunolabeling showed that Kir4.1 and AQP4 were occasionally distributed in close proximity but in distinct compartments of the astroglial cell membrane. Astroglial noncaveolar DRM may therefore be made up of at least two distinct compartments, MbetaCD-sensitive and MbetaCD-resistant microdomains, which control localization and function of, respectively, Kir and AQP4 channels on the cell membrane.


Autogenous mitochondria transplantation for treatment of right heart failure.
Right ventricular hypertrophy and failure are major causes of cardiac morbidity and mortality. A key event in the progression to right ventricular hypertrophy and failure is cardiomyocyte apoptosis due to mitochondrial dysfunction. We sought to determine whether localized intramyocardial injection of autologous mitochondria from healthy muscle treats heart failure. Mitochondria transplanted from different sources were initially tested in cultured hypertrophic cardiomyocytes. A right ventricular hypertrophy/right ventricular failure model created through banding of the pulmonary artery in immature piglets was used for treatment with autologous mitochondria (pulmonary artery banded mitochondria injected/treated n = 6) from calf muscle, versus vehicle (pulmonary artery banded vehicle injected/treated n = 6) injected into the right ventricular free-wall, and compared with sham-operated controls (sham, n = 6). Animals were followed for 8 weeks by echocardiography (free-wall thickness, contractility), and dp/dt max was measured concomitantly with cardiomyocyte hypertrophy, fibrosis, and apoptosis at study end point. Internalization of mitochondria and adenosine triphosphate levels did not depend on the source of mitochondria. At 4 weeks, banded animals showed right ventricular hypertrophy (sham: 0.28 ± 0.01 cm vs pulmonary artery banding: 0.4 ± 0.02 cm wall thickness; P = .001), which further increased in pulmonary artery banded mitochondria injected/treated but declined in pulmonary artery banded vehicle injected/treated (0.47 ± 0.02 cm vs 0.348 ± 0.03 cm; P = .01). Baseline contractility was not different but was significantly reduced in pulmonary artery banded vehicle injected/treated compared with pulmonary artery banded mitochondria injected/treated and so was dp/dtmax. There was a significant difference in apoptotic cardiomyocyte loss and fibrosis in sham versus hypertrophied hearts with most apoptosis in pulmonary artery banded vehicle injected/treated hearts (sham: 1 ± 0.4 vs calf muscle vs vehicle: 13 ± 1.7; P = .001 and vs pulmonary artery banded mitochondria injected/treated: 8 ± 1.9, P = .01; pulmonary artery banded vehicle injected/treated vs pulmonary artery banded mitochondria injected/treated, P = .05). Mitochondrial transplantation allows for prolonged physiologic adaptation of the pressure-loaded right ventricular and preservation of contractility by reducing apoptotic cardiomyocyte loss.


Rotation mechanism of Enterococcus hirae V1-ATPase based on asymmetric crystal structures.
In various cellular membrane systems, vacuolar ATPases (V-ATPases) function as proton pumps, which are involved in many processes such as bone resorption and cancer metastasis, and these membrane proteins represent attractive drug targets for osteoporosis and cancer. The hydrophilic V(1) portion is known as a rotary motor, in which a central axis DF complex rotates inside a hexagonally arranged catalytic A(3)B(3) complex using ATP hydrolysis energy, but the molecular mechanism is not well defined owing to a lack of high-resolution structural information. We previously reported on the in vitro expression, purification and reconstitution of Enterococcus hirae V(1)-ATPase from the A(3)B(3) and DF complexes. Here we report the asymmetric structures of the nucleotide-free (2.8 Å) and nucleotide-bound (3.4 Å) A(3)B(3) complex that demonstrate conformational changes induced by nucleotide binding, suggesting a binding order in the right-handed rotational orientation in a cooperative manner. The crystal structures of the nucleotide-free (2.2 Å) and nucleotide-bound (2.7 Å) V(1)-ATPase are also reported. The more tightly packed nucleotide-binding site seems to be induced by DF binding, and ATP hydrolysis seems to be stimulated by the approach of a conserved arginine residue. To our knowledge, these asymmetric structures represent the first high-resolution view of the rotational mechanism of V(1)-ATPase.


The effect of sacrifice on image signal, T1, T2, and T*2 in liver, kidney, and brain of the Wistar rat.
Signal intensity in SE240/25 and GE240/25 images, T*2, T2, and T1 were measured prior to and after sacrifice at 1.5 T in liver, kidney, and brain of Wistar rats. Presacrifice, animals were maintained on 2-3% halothane, ensuring a high saturation of venous haemoglobin with oxygen, which emphasised blood oxygenation level dependent (BOLD) effects upon deoxygenation of haemoglobin postsacrifice. T*2 values (in ms) changed on sacrifice from 27.8 +/- 4.1 to 15.0 +/- 1.1 in liver (p << .001), from 57.4 +/- 6.5 to 52.8 +/- 5.7 in brain (p < .02), and from 52.3 +/- 10.3 to 37.6 +/- 11.9 in kidney cortex (p < .01). T*2 was also reduced in kidney medulla and papilla. T2 reduced on sacrifice (by a lesser amount than T*2), except in brain, which showed a small increase. T1 (ms) changed only in liver, showing a significant increase from 670 +/- 31 to 834 +/- 69 (p << .001). The ratio of signal postsacrifice to presacrifice was lowest in the SE240/25 and GE240/25 images of liver (0.73 +/- 0.03 and 0.52 +/- 0.05, respectively), but increased T1 may account for some of these changes. Expected signal ratios at sacrifice calculated from measured relaxation times agreed well with those observed experimentally. Relaxation changes observed on sacrifice were consistent with deoxygenation of haemoglobin. These results give an estimate of the maximum effects which changes of blood oxygen level might produce at 1.5 T and suggest that contrast due to variation of endogenous deoxyhaemoglobin may be exploitable in the liver.


Maternal cadmium exposure during late pregnancy causes fetal growth restriction via inhibiting placental progesterone synthesis.
Cadmium (Cd) is a major environmental pollutant. Maternal Cd exposure throughout pregnancy caused fetal growth restriction (FGR). However, the pivotal time window of Cd-evoked FGR and its mechanism are unknown. Here, we will establish a murine model to explore the effects of maternal Cd exposure at different stages of gestation on fetal growth and placental progesterone biosynthesis. Pregnant mice were randomly divided into four groups. For Cd groups, mice were given with CdCl2 (150 mg/L) through drinking water at early (GD0-GD6), middle (GD7-GD12) and late (GD13-GD17) gestation, respectively. The controls received reverses osmosis (RO) water. Results showed that maternal cadmium exposure only in late gestation lowered fetal weight and length. Correspondingly, placental Cd level in late gestational Cd exposure is the highest among three different gestational stages. Although gestational Cd exposure had few adverse effects in the weight and diameter of mouse placenta, placental vascular development, as determined by H&E staining and cluster of differentiation-34 (CD-34) immunostaining, was impaired in mice exposed to Cd during late pregnancy. Additionally, late gestational exposure to cadmium markedly reduced progesterone level in maternal serum and placenta. In line, the expression of key progesterone synthetases, including steroidogenic acute regulatory protein (StAR) and 3β-hydroxyl steroid dehydrogenase (3β-HSD), was obviously downregulated in placenta from mice was exposed Cd during late pregnancy. These data suggest that maternal Cd exposure during late pregnancy, but not early and middle pregnancy, induces fetal growth restriction partially via inhibiting placental progesterone synthesis.


The preparation of leukocyte-poor red cells from AS-1 red cells by the spin-cool-filter method.
Ninety-eight units of AS-1 red cells (RBC) were studied to determine the optimal unit age for the preparation of leukocyte-poor red cells by the spin-cool-filter method. After 1 week of storage, the residual leukocyte counts of AS-1 units prepared by this method averaged 0.5 x 10(9) or fewer white cells, which indicated removal of more than 79 percent of the total white cells. The spin-cool-filter method, in conjunction with the rapid degeneration of the white cells in the AS-1 RBC, yielded leukocyte-poor units comparable to those prepared from citrate-phosphate-dextrose-adenine (CPDA-1) RBC.


Self-reporting of height and weight: valid and reliable identification of malnutrition in preoperative patients.
Preoperative screening for malnutrition has become mandatory in The Netherlands. A sensitive method to diagnose malnutrition would save time and improve effectiveness. A prospective cross-sectional study of 488 adult elective preoperative outpatients was performed. The accuracy of self-reported height and weight was compared with measured data and 3 commonly used malnutrition screening tools. Interobserver agreement was calculated by the intraclass correlation coefficient, studied in Bland and Altman plots, and analyzed by using Cohen's κ statistic. Accuracy was expressed in sensitivity, specificity, and false-negative rates. Differences between self-reported and measured data were significant, but clinically irrelevant, because only 1 patient was falsely identified as well nourished. Intraclass correlation coefficient for height, weight, and body mass index was high (.97-.99). Bland-Altman plots showed that the mean ± standard deviation differences and 95% limits of agreement between both methods were as follows: height, .0096 m (±.0262, -.0417 to +.0609 m); weight, -1.28 kg (±2.29, -5.76 to +3.20 kg); body mass index, -.72 kg/m(2) (±1.11, -2.92 to +1.46 kg/m(2)). The κ coefficient was .84 (95% confidence interval, .75-.94). Sensitivity was .97 and specificity was .98. Sensitivity and false-negative rates of self-reported data were better overall compared with the screening tools. Self-reported data provide highly sensitive information to diagnose malnutrition in preoperative outpatients.


The heat labile phosphatase modulator (inhibitor-2) complex from rabbit skeletal muscle.
The heat stable phosphatase modulator protein (inhibitor-2) has been shown to play a crucial role in the reversible ATP, Mg-dependent activation of a multisubstrate protein phosphatase. The modulator activity is acid and heat stable and resides in a small asymmetrical protein which, after boiling migrates in sucrose density gradient centrifugation with a molecular weight of 17K. The present report shows that in unboiled rabbit skeletal muscle preparations all the modulator activity is found associated with a heat labile protein component, which imposes an important regulatory feature on the heat stable activity. The heat labile complex migrates in sucrose density gradient centrifugation as a Mr = 70K protein.


Comparing Canada's proposed Critical Cyber Systems Protection Act with cybersecurity legal requirements in the EU.
This article examines the Canadian federal government's proposed Critical Cyber Systems Protection Act (CCSPA), compares it with existing and proposed cybersecurity legal requirements in the European Union (EU), and sets out recommendations to address shortcomings of the proposed Canadian legislation. One of the cornerstone components of Bill C‑26, the CCSPA seeks to regulate critical cyber systems in federally regulated private sectors. It represents a significant overhaul of Canadian cybersecurity regulation. However, the current proposed legislation exhibits many flaws, including a commitment to, and entrenchment of, a patchwork approach to regulation that focuses on formal registration; a lack of oversight of its confidentiality provisions; a weak penalty scheme that focuses solely on compliance, not deterrence; and diluted conduct, reporting, and mitigation obligations. To repair these flaws, this article reviews the provisions of the proposed law and compares them with the EU's Directive Concerning Measures for a High Common Level of Security of Network and Information Systems Across the Union, the first EU-wide cybersecurity legislation, as well as its proposed successor, the NIS2 Directive. Where relevant, various other cybersecurity regulations in peer states are discussed. Specific recommendations are put forward.


Expression profile of heptahelical putative membrane progesterone receptors in epithelial ovarian tumors.
A novel class of putative progestin binding proteins has been recently identified as potential mediators of rapid nongenomic hormone actions. The proteins designated membrane progestin receptor (mPR) alpha, beta, and gamma were initially discovered in fish and shown to have a role in oocyte maturation. The predicted multiple membrane spanning domain structure of the mPRs resembles that of heptahelical G-protein-coupled receptors. Phylogenetic analysis indicated that the mPRs belong to the large progestin and adiponectin Q receptor (PAQR) gene family. Based on the reported expression of the 3 mPRs in hormone-responsive tissues of the female reproductive tract and on the role of steroid hormones in cancer, we investigated the expression of these novel progestin receptors in epithelial tumors of the ovary. The transcript levels of the 3 human mPR/PAQRs were assessed by semiquantitative reverse transcriptase polymerase chain reaction in 28 ovarian samples, including normal tissues, cystadenomas, borderline tumors, and common types of ovarian carcinomas. Two of the 3 transcripts for the mPR/PAQRs proteins appeared differentially expressed in the tumors examined. Expression of mPR alpha and beta was demonstrated in ovarian tumors at both messenger RNA and protein level, and their expression appeared to be independent of the expression of the classic nuclear progestin receptors. Expression of mPR gamma (PAQR V) was elevated in endometrioid and clear cell carcinomas, 2 related neoplastic counterparts of hormonally responsive tissues, suggesting a potential role of the mPR/PAQRs in the pathogenesis of epithelial ovarian tumors.


Copper complexation by tannic acid in aqueous solution.
The speciation of titrated copper in a dissolved tannic acid (TA) solution with an initial concentration of 4 mmol organic carbon (OC)/l was investigated in a nine-step titration experiment (Cu/OC molar ratio=0.0030-0.0567). We differentiated between soluble and insoluble Cu species by 0.45 microm filtration. Measurements with a copper ion selective electrode (ISE) and diffusive gradients in thin films (DGT) were conducted to quantify unbound Cu(II) cations ("free" Cu) and labile soluble Cu complexes. For the DGT measurements, we used an APA hydrogel and a Chelex 100 chelating resin (Na form). Insoluble organic Cu complexes (>0.45 microm) was the dominant Cu species for Cu/OC=0.0030-0.0567 with a maximum fraction of 0.96 of total Cu. At Cu/OC>0.0100, Cu-catalysed degradation of aggregate structures resulted in a strong increase of free Cu and (labile) soluble Cu complexes with a maximum fraction of 0.28 and 0.32 of total Cu, respectively. Labile (i.e. DGT-detectable) soluble Cu complexes had a relatively high averaged diffusion coefficient (D) in the APA hydrogel (3.50 x 10(-6)- 5.58 x 10(-6)cm(2)s(-1)).


Interrupted opening statements in clinical encounters: A scoping review.
To conduct a scoping review of literature on the duration of patients' opening statements in clinical encounters, with or without an interruption. We conducted a scoping review to identify articles based on pre-specified inclusion and exclusion criteria. One reviewer extracted study details and outcomes related to the length of patients' opening statements. A second reviewer verified this data. We included 14 studies from 1185 citations reviewed. Encounters were in primary care clinics, specialty clinics, and hospitals. Across six studies, the mean time to interruption was 18.2 s. The mean length of uninterrupted opening statements was 45.9 s across nine studies. Clinicians interrupting their patients' opening statements, compared to not interrupting, takes away time from the patient to fully present their concerns. Research has not focused on the nature of clinician interruptions. For instance, an interruption encouraging expansion or more detail facilitates understanding. Therefore, the impact these interruptions have on the clinical encounter remains unknown. Interrupting the average patient does not save time, so clinicians may benefit from allowing their patients to complete their opening statements.


Morphological and functional findings of fixed phagocytes in psoriatics.
Twenty-one psoriatics as well as 24 normal healthy adults were studied by functional bone marrow scintigraphy using Tc-99m-labeled human serum albumin millimicrospheres (Tc-99m-HSA-MM). Functional bone marrow scintigraphy is an in vivo test system for the assessment of various functional properties of fixed phagocytes. Of psoriatics who had no systemic drug treatment, 59% demonstrated peripheral extension of the bone marrow space, indicating hyperplasia of bone marrow phagocytes. This phenomenon could be observed only in one normal subject who was a high-performance sportsman. Of psoriatics treated with aromatic retinoid, 83% (n = 6) demonstrated bone marrow extension, as did 100% (n = 3) of psoriatics with cirrhosis of liver. The "capacity' of bone marrow phagocytes to engulf Tc-99m-HSA-MM ("uptake ratio') was diminished in 34% of nontreated as well as 66% of psoriatics treated with aromatic retinoid. The phagocytic and proteolytic turnover of Tc-99m-HSA-MM in the bone marrow, spleen, and liver was found to e accelerated in 66% of nontreated psoriatics, normal (83%) or accelerated (17%) in psoriatics treated with aromatic retinoid, and considerably delayed in all of the psoriatics with cirrhosis of liver. Functional bone marrow scintigraphy proved to be an appropriate in vivo test system for revealing abnormalities of fixed phagocytes in psoriatics. Furthermore, therapeutic effects as well as the influences of preexisting disorders on different phagocyte populations can be assessed.


Proliferative response of T cells from tumor-immune chickens to carcinogen-induced fibrosarcoma cells.
Immunity to carcinogen-induced transplantable fibrosarcomas (CHCT-NYU-4, -97, -36 and -20) in SC chickens was studied by the ability of spleen cells from NYU-4 or -97 immune chickens to proliferate in response to tumor cells in vitro. Spleen, but not peripheral blood cells, from NYU-4 immune chickens proliferated significantly more vigorously to gamma-irradiated NYU-4 cells than did cells from normal chickens. The proliferative response was not much affected by addition of indomethacin. Spleen cells from NYU-4-immune agammaglobulinemic (A gamma) chickens exhibit the same ability to proliferate in presence of gamma-irradiated NYU-4 tumor cells. Analysis of the phenotype of the T-cell component involved in proliferation showed that the proliferative response was significantly decreased by removal of CT4+ cells through indirect panning. Removal of CT8+ cells enhanced background proliferation without affecting the total thymidine incorporation in the presence of tumor cells. Immune spleen cells usually gave highest responses to the immunizing tumor, but also exhibited cross-reactivity to cells from other individual tumors induced by the same carcinogen.


Stages in the delivery of home-based services to parents at risk of child abuse: a Healthy Start experience.
Hawaii's Healthy Start program of home-based services to families identified at risk for child abuse and neglect is becoming a model for the nation. The present paper provides background information about Healthy Start and describes a three-phase process of engagement and service delivery that characterizes the Healthy Start home visitation model.


Motion-compensated low-rank reconstruction for simultaneous structural and functional UTE lung MRI.
Three-dimensional UTE MRI has shown the ability to provide simultaneous structural and functional lung imaging, but it is limited by respiratory motion and relatively low lung parenchyma SNR. The purpose of this paper is to improve this imaging by using a respiratory phase-resolved reconstruction approach, named motion-compensated low-rank reconstruction (MoCoLoR), which directly incorporates motion compensation into a low-rank constrained reconstruction model for highly efficient use of the acquired data. The MoCoLoR reconstruction is formulated as an optimization problem that includes a low-rank constraint using estimated motion fields to reduce the rank, optimizing over both the motion fields and reconstructed images. The proposed reconstruction along with XD and motion state-weighted motion-compensation (MostMoCo) methods were applied to 18 lung MRI scans of pediatric and young adult patients. The data sets were acquired under free-breathing and without sedation with 3D radial UTE sequences in approximately 5 min. After reconstruction, they went through ventilation analyses. Performance across reconstruction regularization and motion-state parameters were also investigated. The in vivo experiments results showed that MoCoLoR made efficient use of the data, provided higher apparent SNR compared with state-of-the-art XD reconstruction and MostMoCo reconstructions, and yielded high-quality respiratory phase-resolved images for ventilation mapping. The method was effective across the range of patients scanned. The motion-compensated low-rank regularized reconstruction approach makes efficient use of acquired data and can improve simultaneous structural and functional lung imaging with 3D-UTE MRI. It enables the scanning of pediatric patients under free-breathing and without sedation.


Altered glycosylation of proteins produced by malignant cells, and application for the diagnosis and immunotherapy of tumours.
Most secretory and membrane-bound proteins produced by mammalian cells contain covalently linked sugar chains. Alterations of the sugar chain structures of glycoproteins have been found to occur in various tumours. Because the sugar chains of glycoproteins are essential for the maintenance of the ordered social behaviour of differentiated cells in multicellular organisms, alterations to the sugar chains are the molecular basis of abnormal social behaviours in tumour cells, such as invasion into the surrounding tissues and metastasis. In this review, the structure and enzymatic basis of typical alterations of the N-linked sugar chains, which are found in various tumours, are introduced. These data are useful for devising diagnostic methods and immunotherapies for the clinical treatment of tumours. Three beta-N-acetylglucosaminyltransferases, GnT-III, -IV and -V, play roles in the structural alteration of the complex-type sugar chains in various tumours. In addition, transcriptional changes in various glycosyltransferases, together with the transporters of sugar nucleotides and sulfate, which are responsible for the formation of the outer chain moieties of complex-type sugar chains, are the keys to inducing the alterations.


A comparative study on the formation and characterization of aerobic 4-chloroaniline-degrading granules in SBR and SABR.
The formation and characterization of the aerobic 4-chloroaniline-degrading granules in the three column-type sequencing batch reactors were investigated in this paper. The granular sludge was observed since 15 days after start-up in R2 and R3 which had the high ratio of height to diameter (H/D). Since then and within the subsequent 75 days, the granulation of aerobic sludge was apparently developed by the decreased settling time and gradually increased 4-chloroaniline (4-ClA) concentration to above 400 mg.L(-1) in R1 to R3. The aerobic granules tended to be mature in all reactors continuously operated with 4-ClA loading rates of around 800 g.m(-3).d(-1), and the removal efficiencies of chemical oxygen demand, total nitrogen, and 4-ClA were maintained above 93%, 70%, and 99.9%, respectively. Mature aerobic granules in R1 to R3 featured with the average diameter of 0.78, 1.68, and 1.25 mm, minimal settling velocity of 20.5, 70.1 and 66.6 m.h(-1), specific 4-ClA degradation rates of 0.14, 0.21, and 0.27 g.gVSS(-1).d(-1), and the ratio of proteins to polysaccharides of 8.2, 10.8, and 13.7 mg.mg(-1), respectively. This study demonstrates that the reactor with a high H/D ratio and internal circulation favors the granulation and stabilization of aerobic sludge.


Migration of a Silastic central venous catheter into the right jugular vein: report of two cases.
Silastic catheters commonly used for total parenteral nutrition have advantages including low rates of infection and thrombosis. However, because of flexibility they produce an unusual problem, catheter tip migration. We present two cases of catheter migration into the right jugular vein. In one case, infusion phlebitis developed which required repositioning over a guidewire, and in a second a spontaneous correction occurred.


Graphical representation of QT rate correction formulae: an aid facilitating the use of a given formula and providing a visual comparison of the impact of different formulae.
The QT interval on the electrocardiogram is an increasingly important measurement, especially in relation to drug action and interaction. The QT interval varies inversely as the heart rate and numerous rate correction formulae have been proposed. It is difficult to compare the effect of applying different formulae at different heart rates and for different measured QT intervals. A simple graphical display of the results from different formulae is proposed. This display is dependent on the concept of the absolute correction factor. This graphical presentation is useful (a) in comparing the effect of the application of different formulae and (b) in directly reading the correction produced by any individual formula.


Dysphagia in stroke and neurologic disease.
Dysphagia is a common problem in neurologic disease. The authors describe rates of dysphagia in selected neurologic diseases, and the evaluation and treatment of dysphagia in this population. Applicable physiology and aspects of neural control are reviewed. The decision-making process to determine oral feeding versus alternative means of alimentation is examined.


Development of odour awareness in pre-schoolers: A longitudinal study.
Conducting interviews about children's olfactory behaviours offers a feasible way of learning about the earliest perceptions and knowledge of one's odour world. However, little is known about the stability and development of such self-reports. Here we present the first longitudinal study to repeatedly test children's odour awareness using the Children's Olfactory Behavior in Everyday Life (COBEL) questionnaire in five waves over a two-year period. We expected that higher scores would be attained by girls relative to boys and by older children compared to younger ones and that the scores would increase further into the study. We found a linear time-related increase in the total COBEL scores and in the food and environmental components, whereas awareness of social odours decreased over time. Girls were more olfaction-oriented in the context of social and environmental, but not food, odours. All the reported effects were small. The age at which the children entered the study did not affect their scores. We suggest that the unexpected findings regarding social odours warrant replication in particular and extension in longitudinal studies carried out over a broader time span.


Early Improvement with Vortioxetine Predicts Response and Remission: A Post Hoc Analysis of Data from a Clinical Trial Conducted in Japan.
Several weeks of treatment with an antidepressive agent may be required before efficacy is demonstrated in patients with major depressive disorder. This study investigated the predictive value of early partial improvement with vortioxetine for treatment response and remission. This was a post hoc analysis of an 8-week, randomized, double-blind, placebo-controlled, Phase 3 study of vortioxetine (10 mg or 20 mg) in Japanese patients aged 20-75 years with recurrent major depressive disorder and a Montgomery-Åsberg Depression Rating Scale (MADRS) score of at least 26. The key outcomes were the predictive value of early partial improvement (reduction in MADRS total score of ≥20% from baseline to week 2) with vortioxetine for MADRS response (≥50% decrease in score from baseline) and remission (decrease in score to ≤10) at week 8. Relevant data were available for 478 patients; 62/158 patients receiving placebo, 71/162 receiving vortioxetine 10 mg, and 66/158 receiving vortioxetine 20 mg were early improvers. Early improvers receiving vortioxetine (10 mg or 20 mg) were more likely than non-early improvers to achieve a week 8 response (71.2-73.2% vs 29.7-38.0%) or remission (50.7-51.5% vs 17.4-18.7%). Positive predictive values for response and remission with vortioxetine were ~70% and ~50%, respectively; negative predictive values were ~70% and ~80%, respectively. Improvement with vortioxetine may be predicted by early partial improvement in MADRS score. Some patients may benefit from longer-term treatment even without early improvement, another finding that may aid clinical decision-making. ClinicalTrials.gov registration for primary study: NCT02389816.


The arc of Buhler: special considerations when performing pancreaticoduodenectomy.
A 74-year-old female was diagnosed as having a carcinoma of the papilla of Vater. Preoperative computed tomography showed stenosis of the celiac trunk and an enlarged artery arising from the superior mesenteric artery (SMA) joining the root of the splenic artery. Since this artery communicated with the SMA and the celiac trunk, independently of the gastroduodenal and dorsal pancreatic arteries, it was considered to be the arc of Buhler (AOB). The arterial blood flow to the liver, spleen, and stomach appeared to depend on the AOB, such that AOB preservation was considered to be essential. A subtotal stomach-preserving pancreaticoduodenectomy with preservation of the AOB was thus performed. Although AOB is a relatively infrequent type of arterial communication between the SMA and the celiac trunk, it needs to be preserved during pancreaticoduodenectomy when celiac trunk stenosis is present.


Epithelial-specific ETS-1 (ESE1/ELF3) regulates apoptosis of intestinal epithelial cells in ulcerative colitis via accelerating NF-κB activation.
Epithelial-specific ETS-1 (ESE1), also named as ELF3, ERT and ESX, belonging to the ETS family of transcription factors, exerts multiple activities in inflammation, epithelial differentiation and cancer development. Previous data demonstrated that ESE1 synergizes with NF-κB to induce inflammation and drive tumor progress, and the nuclear translocation of ESE1 promotes colon cells apoptosis. However, the expression and biological functions of ESE1 in ulcerative colitis (UC) remain unclear. In this study, we reported for the first time that ESE1/ELF3 was over-expressed in intestinal epithelial cells (IECs) of patients with UC. In DSS-induced colitis mouse models, we observed the up-regulation of ESE1/ELF3 accompanied with the elevated levels of IEC apoptotic markers (active caspase-3 and cleaved PARP) and NF-κB activation indicators [phosphorylated NF-κB p65 subunit (p-p65) and p-IκB] in colitis IECs. Increased co-localization of ESE1/ELF3 with active caspase-3 (and p-p65) in IECs of the DSS-induced colitis group further indicated the possible involvement of ESE1/ELF3 in NF-κB-mediated IEC apoptosis in UC. Employing the TNF-α-treated HT-29 cells as an IEC apoptosis model, we confirmed the positive correlation of ESE1/ELF3 with NF-κB activation and caspase-dependent IEC apoptosis in vitro. Immunoprecipitation and immunofluorescence assay revealed the physical interaction and increased nuclear translocation of ESE1/ELF3 and the NF-κB p65 subunit in TNF-α-treated HT-29 cells. Knocking ESE1/ELF3 down by siRNA significantly alleviated TNF-α-induced NF-κB activation and cellular apoptosis in HT-29 cells. Taken together, our data suggested that ESE1/ELF3 may promote the UC progression via accelerating NF-κB activation and thus facilitating IEC apoptosis.


Biological reference values for chemical compounds in the work area (BARs): an approach for evaluating biomonitoring data.
Biological monitoring is a routine method that has been applied in occupational medical practice for many years. A requirement for its application is the availability of criteria suitable for evaluating biomonitoring data. Health-based threshold values are particularly useful as a criterion, yet only for substances for which effect thresholds can reliably be determined. For substances for which the concept of health-based threshold values is not applicable, the Working Group Setting of Threshold Limit Values in Biological Materials of the DFG Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area has recently established "Biologische Arbeitsstoff-Referenzwerte" (BARs, Biological Reference Values for Chemical Compounds in the Work Area) as an approach for evaluating biomonitoring data. The BAR represents the upper reference concentration of a biomarker in the general adult population without occupational exposure to the agent. It is derived from biomonitoring data of a sample of a defined population group. In general, a BAR corresponds to the 95th percentile of the sample distribution. Ideally, national environmental surveys including human biomonitoring results are used as basis for deriving BARs. The influence of age, sex, social status, residential area and life style factors on background exposure is considered in the evaluation of these values. Because tobacco smoking is the most frequent influencing factor, several BARs have been determined for non-smokers only. To date, BARs for 17 substances or substance groups are listed in the List of MAK and BAT Values 2011. BARs for another five substances have been discussed, but have not been established because of the insufficient scientific database. Establishing the BARs aims to facilitate the evaluation of human exposure to chemical compounds for which no health-based threshold values can be derived but an adequate assessment of exposure is required due to their toxicity. The application of BARs does not permit a toxicological evaluation, but does allow the occurrence and the extent of occupational exposure to hazardous substances to be proved.


Kaiser Permanente's performance improvement system, Part 4: Creating a learning organization.
In 2006, recognizing variations in performance in quality, safety, service, and efficiency, Kaiser Permanente leaders initiated the development of a performance improvement (PI) system. Kaiser Permanente has implemented a strategy for creating the systemic capacity for continuous improvement that characterizes a learning organization. Six "building blocks" were identified to enable Kaiser Permanente to make the transition to becoming a learning organization: real-time sharing of meaningful performance data; formal training in problem-solving methodology; workforce engagement and informal knowledge sharing; leadership structures, beliefs, and behaviors; internal and external benchmarking; and technical knowledge sharing. Putting each building block into place required multiple complex strategies combining top-down and bottom-up approaches. Although the strategies have largely been successful, challenges remain. The demand for real-time meaningful performance data can conflict with prioritized changes to health information systems. It is an ongoing challenge to teach PI, change management, innovation, and project management to all managers and staff without consuming too much training time. Challenges with workforce engagement include low initial use of tools intended to disseminate information through virtual social networking. Uptake of knowledge-sharing technologies is still primarily by innovators and early adopters. Leaders adopt new behaviors at varying speeds and have a range of abilities to foster an environment that is psychologically safe and stimulates inquiry. A learning organization has the capability to improve, and it develops structures and processes that facilitate the acquisition and sharing of knowledge.


Differential diagnosis of small cell glioblastoma and anaplastic oligodendroglioma: a case report of an elderly man.
Small cell glioblastoma is a histological subtype of glioblastoma with characteristic features of highly proliferative, monotonous small glial cells with high nuclear cytoplasm ratio. Morphologically, malignant lymphoma or small cell metastatic carcinoma should be carefully discriminated. Some cases are difficult to differentiate from anaplastic oligodendroglioma. In this report, we present a case of small cell glioblastoma of an elderly man. The lack of IDH1/2 mutation was confirmed by immunohistochemistry and direct sequencing. Fluorescence in situ hybridization revealed the lower rates of chromosome 1p and 19q deletion. Microsatellite analysis disclosed partial 10q alteration near the PTEN locus. Not only morphological and immunohistochemical examinations, but also cytogenetical investigations for IDH1/2 mutation, 1p/19q loss, and PTEN alteration, are strongly supportive methods for the differential diagnosis of small cell glioblastoma and anaplastic oligodendroglioma.


The t(3;21) fusion product, AML1/Evi-1 blocks AML1-induced transactivation by recruiting CtBP.
AML1/Evi-1 is a chimeric protein that is derived from t(3;21), found in blastic transformation of chronic myelogenous leukemia. It is composed of the N-terminal AML1 portion with the DNA-binding Runt domain and the C-terminal Evi-1 portion. It has been shown to dominantly repress AML1-induced transactivation. The mechanism for it has been mainly attributed to competition with AML1 for the DNA-binding and for the interaction with PEBP2beta (CBFbeta), a partner protein which heterodimerizes with AML1. It was recently found that Evi-1 interacts with C-terminal binding protein (CtBP) to repress TGFbeta-induced transactivation. Here, we demonstrate that AML1/Evi-1 interacts with CtBP in SKH1 cells, a leukemic cell line which endogenously overexpresses AML1/Evi-1 and that AML1/Evi-1 requires the interaction with CtBP to repress AML1-induced transactivation. The association with CtBP is also required when AML1/Evi-1 blocks myeloid differentiation of 32Dcl3 cells induced by granulocyte colony-stimulating factor. Taken together, it is suggested that one of the mechanisms for AML1/Evi-1-associated leukemogenesis should be an aberrant recruitment of a corepressor complex by the chimeric protein.


Boesenbergia rotunda extract accelerates human keratinocyte proliferation through activating ERK1/2 and PI3K/Akt kinases.
Boesenbergia rotunda (BR) has long been used as tradition medicine. For its pharmacological effects on wound healing, previous studies in an animal model provided convincing results that the ethanolic extract from the rhizome of this plant can stimulate wound healing. However, the mechanism about how this plant promotes wound healing at the molecular level has not been elucidated. As a step towards the development of wound healing agents, our current study utilized a human keratinocyte cell line (HaCaT) as an in vitro model to define the potential molecular mechanisms of BR extract in enhancing wound-healing. Our HPLC results showed that BR extract contained kaempferol as one of its potential compounds. The extract strongly promoted wound healing of HaCaT cell monolayer. This effect was eventually defined to be regulated through the ability of BR extract to induce cell proliferation. At the signaling level, we discovered that BR extract rapidly activated ERK1/2 and Akt phosphorylation upon the addition of the extract. Additionally, our experiments where specific inhibitors of MEK (U0126) and PI3K (LY294002) were utilized verified that BR enhanced cell proliferation and wound healing through stimulating the MAPK and PI3K/Akt signal transduction pathways. Moreover, direct inhibition of keratinocyte DNA synthesis by mitomycin C (MMC) could completely block the proliferative effects of BR extract. Nevertheless, data from Transwell migration assay revealed that BR extract did not promote keratinocyte migration. Altogether, we provided more evidence that BR possesses its wound healing-promoting action through the activation of proliferation and survival pathways, and our study suggests that BR is an interesting candidate to be developed as a wound healing-promoting agent.


Secreted protein eco-corona mediates uptake and impacts of polystyrene nanoparticles on Daphnia magna.
Nanoparticles (NPs) are defined as having at least one external dimension between 1 and 100 nm. Due to their small size, NPs have a large surface area to volume ratio giving them unique characteristics that differ from bulk material of the same chemical composition. As a result these novel materials have found numerous applications in medical and industrial fields with the result that environmental exposure to NPs is increasingly likely. Similarly, increased reliance on plastic, which degrades extremely slowly in the environment, is resulting in increased accumulation of micro-/nano-plastics in fresh and marine waters, whose ecotoxicological impacts are as yet poorly understood. Although NPs are well known to adsorb macromolecules from their environment, forming a biomolecule corona which changes the NP identity and how it interacts with organisms, significantly less research has been performed on the ecological corona (eco-corona). Secretion of biomolecules is a well established predator-prey response in aquatic food chains, raising the question of whether NPs interact with secreted proteins, and the impact of such interaction on NP uptake and ecotoxicity. We report here initial studies, including optimisation of protocols using carboxylic-acid and amino modified spherical polystyrene NPs, to assess interaction of NPs with biomolecules secreted by Daphnia magna and the impact of these interactions on NP uptake, retention and toxicity towards Daphnia magna. Daphnia magna are an important environmental indicator species who may be especially sensitive to nanoparticles (NPs) as a result of being filter-feeders. This paper demonstrates for the first time that proteins released by Daphnia magna create an eco-corona around polystyrene NPs which causes heightened uptake of the NPs and consequently increases toxicity. The secreted protein eco-corona also causes the NPs to be less efficiently removed from the gut of D. magna and NPs remaining in the gut of D. magna affected the rate of subsequent feeding. Thus, fate of NPs in the environment should be evaluated and monitored under more realistic exposure scenarios.


Midwifery workplace culture in Australia: A national survey of midwives.
The midwifery workforce in Australia is impacted by shortages and attrition. Workplace culture affects midwives' intentions to stay in the profession and their capacity to provide woman-centred care for mothers and infants. Staff attrition in maternity services often relates to midwives' workplace experiences and negative perceptions of organisational culture. Broad-based data are essential to fully understand midwifery workplace culture. This study aimed to examine Australian midwives' perceptions of workplace culture, using a specifically developed instrument. A national online survey of Australian midwives, within a wider project on maternity workplace culture. Quantitative data were analysed descriptively. Qualitative data were analysed using content analysis. Overall, 322 eligible midwives rated workplace culture and 150 provided further qualitative responses. Themes included 'the ability to be a midwife', 'support at work' and 'bullying'. Less than a third of midwives thought their workplace had a positive culture. Many respondents felt disengaged and unsupported by managers and described an inability to use all their midwifery knowledge in medically-dominated environments. Many attributed poor workplace culture to limited resources, poor communication, time pressure and a lack of leadership in their workplaces. Inadequate staffing levels and poor management left many midwives feeling disempowered and despondent about their workplace. Others, however, described highly positive workplace cultures and inspiring role models. The survey captured a snapshot of Australian midwifery workplace culture. Findings on leadership, workloads, management support and other aspects of workplace culture can inform future workforce planning and policies. A larger study of the midwifery workplace culture is needed.


Norepinephrine induced epithelial-mesenchymal transition in HT-29 and A549 cells in vitro.
Norepinephrine (NE) has been implicated in epithelial-mesenchymal transition (EMT) of cancer cells. However, the underlying mechanism is poorly understood. The goal of this study was to explore the effect of NE on cancer cell EMT and to investigate the potential mechanism. HT-29 and A549 cells were treated with NE, β-adrenergic receptor (β-AR) antagonist (propranolol) or inhibitor of transforming growth factor-β (TGF-β) receptor type I kinase (Ly2157299). Morphology of cells was observed with optical and electron microscope and immunofluorescence staining. Cellular migration and invasion were tested with transwell migration assay and Matrigel invasion assay, respectively. TGF-β1 and cyclic adenosine monophosphate (cAMP) were quantified. EMT markers and signaling pathway were measured by RT-PCR and western blot. NE stimulated TGF-β1 secretion and intracellular cAMP synthesis, induced morphological alterations in HT-29 and A549 cells, and enhanced their ability of migration and invasion. EMT markers induction was observed in NE-treated cancer cells. The effect of NE could be inhibited by propranolol or Ly2157299. β-AR/TGF-β1 signaling/p-Smad3/Snail and β-AR/TGF-β1 signaling/HIF-1α/Snail were two signaling pathways. These findings demonstrated that TGF-β1 signaling pathway was a significant factor of NE-induced cancer cells EMT. The data also suggested that psychological stress might be a risk factor which enhances the ability of migration or invasion of cancer cells.


Ambulatory and in-hospital continuous recording of sleep state and cardiorespiratory parameters in 'near miss' for the sudden infant death syndrome and control infants.
Electrophysiologic and cardiorespiratory events were studied polygraphically and by ambulatory monitoring in 'near miss' for sudden infant death syndrome (SIDS) and normal control infants 1-4 months old. The data show that 'near miss' babies typically have many apneic periods in their sleep, particularly apneas lasting 10 sec or longer. These tend to decrease with age. They have longer lasting apneas than controls, although the latter have respiratory pauses lasting less than 10 sec, in particular during rapid eye movement (REM) sleep. An upper respiratory infection (URI) in 'near miss' infants clearly appears to be a risk factor which will be studied further both in-hospital and at home using a monitoring technique with our Medilog ambulatory system.


Experimental designs and brain mapping approaches for studying the placebo analgesic effect.
The placebo effect has intrigued scientists since it was proposed. The debate has now centered on how it works. Significant progress has been made and most of our knowledge about the neurobiological mechanisms comes from the field of pain and analgesia. The appropriateness of the experimental/clinical paradigms is crucial when we want to investigate the mechanisms of the placebo phenomenon. Recently, functional imaging techniques, such as positron emission tomography, magnetic resonance imaging, and electro/magnetoencephalography have also given the opportunity to define the neuroanatomical bases of placebo analgesia. This work systematically reviews the literature that deals with placebo analgesia, emphasizing both the methodological aspects and the neurobiological advances. The understanding of placebo mechanisms is fundamental and necessary to identify ways of accessing and harnessing these mechanisms in clinical practice to the patient's benefit.


A Surgeon's Guide to Treating Older Patients with Colorectal Cancer.
Review strategies to improve outcomes of colorectal cancer treatment in older patients. Older colorectal patients face many barriers to recovery during their initial inpatient stay following surgery and after leaving the hospital. In addition to the risk of inpatient morbidity and mortality, older patients are more likely to require post-acute care services, to face nutritional deficits, and to experience complications of chemoradiation. In order to improve outcomes for older patients with colorectal cancer, it is important for surgeons to recognize their unique needs and to develop plans to address them. The involvement of a multidisciplinary team with geriatric experience can guide planning for surgery, the immediate postoperative recovery, and long-term survivorship.


First Molecular Identification of Ancylostoma Species in Dogs in a Rural Tea Estate Community in Sri Lanka and the Detection of Other Zoonotic Gastro-intestinal Parasites.
Ancylostoma spp. causing canine hookworm infections are distributed worldwide with a high prevalence in the South Asian region. Here, we report the first molecular identification of the canine hookworm infecting the dogs in Sri Lanka. Fresh faecal samples were collected from free-roaming dogs in tea estate communities in Talawakelle, processed and microscopically analysed for gastro-intestinal parasites. The common dog hookworm Ancylostoma sp. was further investigated using molecular diagnostic tools including PCR, sequencing, and phylogenetic analyses. Of the individual dogs sampled, 97.0% (97/100) were positive for GI parasites, identifying 14 species, all of which were zoonotic. Entamoeba sp. (58.0% (58/100)) was the most prevalent infection and Ancylostoma sp. (45.0% (45/100)) was the most prevalent helminth infection. Overall, helminth prevalence was higher in female dogs (92.5% (37/40)) than male dogs (73.3% (44/60)), potentially due to immune suppression during pregnancy and lactation. The prevalence and burden of Toxocara canis was higher in puppies (≤ 1 year) (68.8%(11/16); Chi-square test; χ2 = 20.912, p < 0.001 and FEC = 5.91; Mann-Whitney U test; p < 0.001), possibly due to vertical trans-mammary or trans-placental transmission of infective larvae. This study also presents the first record of Hymenolepis sp. from dogs in Sri Lanka. Ancylostoma sp. was molecularly confirmed as A. caninum. The local variant of A. caninum was phylogenetically unique and showed 99.2% similarity to that from India. The high prevalence of zoonotic GI parasites with public health significance highlights the importance of improving community well-being, veterinary care for dogs, and public awareness on zoonoses.


Extraction and quantitation of total cholesterol, dolichol and dolichyl phosphate from mammalian liver.
A procedure is described for the determination of total cholesterol, dolichol and dolichyl phosphate (Dol-P) in mammalian liver. It is based on extraction of these compounds into diethyl ether after alkaline saponification of the tissue. Extractability is affected by the length of saponification and concentration of potassium hydroxide (KOH) in the saponification mixture. After extraction, total cholesterol and dolichol are quantitated directly by reverse-phase high pressure liquid chromatography (HPLC) on C18. Dol-P requires further purification before quantitation by HPLC, this is accomplished by chromatography on silicic acid. These methods gave recoveries of over 90% for cholesterol and dolichol and about 60% for Dol-P, using [4-14C]cholesterol, a polyprenol containing 15 isoprene units, and [1-14C]Dol-P as recovery standards. Concentrations of total cholesterol, dolichol and Dol-P in livers from one month-old-CBA mice were found to be 5.7 +/- 0.7 mg/g, 66.3 +/- 1.2 micrograms/g and 3.7 +/- 0.3 micrograms/g, respectively.


Integrin specificity and enhanced cellular activities associated with surfaces presenting a recombinant fibronectin fragment compared to RGD supports.
Biomimetic strategies focusing on presenting short bioadhesive oligopeptides, including the arginine-glycine-aspartic acid (RGD) motif present in numerous adhesive proteins, on a non-fouling support have emerged as promising approaches to improve cellular activities and healing responses. Nevertheless, these bio-inspired strategies are limited by low activity of the oligopeptides compared to the native ligand due to the absence of complementary or modulatory domains. In the present analysis, we generated well-defined biointerfaces presenting RGD-based ligands of increasing complexity to directly compare their biological activities in terms of cell adhesion strength, integrin binding and signaling. Mixed self-assembled monolayers of alkanethiols on gold were optimized to engineer robust supports that present anchoring groups for ligand tethering within a non-fouling, protein adsorption-resistant background. Controlled bioadhesive interfaces were generated by tethering adhesive ligands via standard peptide chemistry. On a molar basis, biointerfaces functionalized with the FNIII7-10 recombinant fragment presenting the RGD and PHSRN adhesive motifs in the correct structural context exhibited significantly higher adhesion strength, FAK activation, and cell proliferation rate than supports presenting RGD ligand or RGD-PHSRN, an oligopeptide presenting these two sites separated by a polyglycine linker. Moreover, FNIII7-10-functionalized surfaces displayed specificity for alpha5beta1 integrin, while cell adhesion to supports presenting RGD or RGD-PHSRN was primarily mediated by alphavbeta3 integrin. These results are significant to the rational engineering of bioactive materials that convey integrin binding specificity for directed cellular and tissue responses in biomedical and biotechnological applications.


Tissue-water relations of two co-occurring evergreen Mediterranean species in response to seasonal and experimental drought conditions.
Tissue-water relations were used to characterize the responses of two Mediterranean co-occurring woody species (Quercus ilex L. and Phillyrea latifolia L.) to seasonal and experimental drought conditions. Soil water availability was reduced approximately 15% by partially excluding rain throughfall and lateral flow (water runoff). Seasonal and experimental drought elicited physiological and morphological adaptations other than osmotic adjustment: both species showed large increases in cell-wall elasticity and decreased saturated-to-dry-mass ratio. Increased elasticity (lower elastic modulus) resulted in concurrent decreases in relative water content at turgor loss. In addition, P. latifolia showed significant increases in apoplastic water fraction. Decreased saturated-to-dry-mass ratio and increased apoplastic water fraction were accompanied by an increased range of turgor maintenance, which indicates that leaf sclerophyllous traits might be advantageous in drier scenarios. In contrast, the degree of sclerophylly (as assessed by the leaf mass-to-area ratio) was not related to tissue elasticity. An approximately 15% reduction in soil water availability resulted in significant reductions in diameter growth when compared to control plants in both species. Moreover, although P. latifolia underwent larger changes in tissue water-related traits than Q. ilex in response to decreasing water availability, growth was more sensitive to water stress in P. latifolia than in Q. ilex. Differences in diameter growth between species might be partially linked to the effects of cell-wall elasticity and turgor pressure on growth, since Q. ilex showed higher tissue elasticity and higher intrinsic tolerance to water deficit (as indicated by lower relative water content at turgor loss) than P. latifolia.


Newborn Hearing Screening with Two-Step Protocol and Risk Factor Identification: Our Experience at a Tertiary Care Centre in Eastern India.
To estimate the prevalence of hearing loss and identify the high-risk factors among neonates with hearing loss. Retrospective study done on 1054 infants in a tertiary care centre in Eastern India from 2020 to 2021 and approved by the Institutional Ethics Committee. A two-step protocol is used for screening. In the well-nursed group, OAE and BOA were performed. In the case of REFER results for automated ABR following OAE evaluation in well-nursed babies, a detailed audiological evaluation was scheduled to be carried out using diagnostic ABR within one month of age. In the high-risk group, hearing screening includes OAE, BOA, and AABR evaluations. AABR evaluation was performed as a part of the screening protocol irrespective of the results of OAE screening as PASS or REFER. In our study among 1053 neonates screened, 375 were in the risk category, and 679 were without risk factors. The overall prevalence of hearing loss in neonates was 22.78 per 1000 screened neonates and 56 per 1000 among high-risk neonates. In the high-risk group, we were able to identify 4 cases of Auditory spectrum neuropathy disorder with the use of AABR during 1st step of screening. In multivariate regression analysis, the risk factors for hearing loss identified were NICU stay (OR = 3.6, 95% CI = 1.1-12.03) and Craniofacial anomalies (OR = 55.37, 95% CI = 16.48- 186.01). Early neonatal screening helps in the detection, intervention, and rehabilitation of hearing loss. The use of AABR in risk infant screening enhanced the chance of detection of auditory spectrum neuropathy disorder (ASND) cases. The online version contains supplementary material available at 10.1007/s12070-023-03723-3.


Enzymatic diagnosis of medium-chain acyl-CoA dehydrogenase deficiency by detecting 2-octenoyl-CoA production using high-performance liquid chromatography: a practical confirmatory test for tandem mass spectrometry newborn screening in Japan.
Many of the previously described enzymatic assay methods for the diagnosis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency have been dependent upon the measurement of radioisotope-labeled co-products or reduction of electron acceptors. We have developed a direct assay method to detect 2-enoyl-CoA production using high-performance liquid chromatography (HPLC). Crude cell lysate prepared from lymphocytes were incubated with n-octanoyl-CoA and ferrocenium hexafluorophosphate. The detection of 2-octenoyl-CoA was significantly reproducible. We applied the assay to samples from four infants suspected to have MCAD deficiency by tandem mass spectrometry (MS/MS) newborn screening conducted in the Hiroshima area of Japan. Three of them were proved to have pathologically reduced residual enzyme activities, although they were associated with various clinical and biochemical phenotypes. In addition, another symptomatic Japanese patient and her presymptomatic sibling who were detected by MS/MS selective screening were successfully diagnosed by our enzymatic assay. These results indicate that the method can be a useful confirmatory test for MS/MS screening of MCAD deficiency.


Synthesis and biological activity of novel acridinylidene and benzylidene thiazolidinediones.
A novel set of acridinylidene thiazolidinediones and benzylidene thiazolidinediones was synthesized by nucleophilic addition of cyanoacrylates. Some of these compounds were evaluated for their glucose lowering capability and their effects on the triglyceride level in alloxan diabetic mice.


Cost-effectiveness of modern mTOR inhibitor based immunosuppression compared to the standard of care after renal transplantation in Germany.
Standards of immunosuppression in renal transplantation have changed dynamically in recent years. We here provide a refined advanced pharmacoeconomic model which uses state-of-the-art methods including a mixed treatment comparison (MTC) analysis. The aim was to assess the cost-effectiveness of current immunosuppressive therapy regimens (TR): "sirolimus + early withdrawal of cyclosporine + steroids" (TR1), "sirolimus-early transition" (TR2), "everolimus-early transition" (TR3) and "tacrolimus low dose + mycophenolate mofetil (MMF) + steroids" (TR4). An up-to-date Markov model with current source data was employed to assess the cost-effectiveness of modern immunosuppressive regimens over 12-month and 10-year time periods. Transition probabilities for the occurrence of events for the first year were based on an MTC analysis. The robustness of the model was tested in extensive sensitivity analyses. Within the 12-month time period TR2 yields the highest life years (0.987 LY), generating costs of 17,500 <euro>. In terms of years with functioning graft (FG), TR4 yields the best efficacy over the 12-month model duration (0.970 years with FG). For the 10-year time period, TR2 yields the lowest costs (107,246 <euro>) and dominates both TR3 and TR1, as it is simultaneously more effective. Within the 10-year model duration, TR4 reaches slightly higher effects compared with TR2 (6.493 vs. 6.474 LY) resulting in an incremental cost-effectiveness ratio of 387,684 <euro> per LY gained. The early transition to sirolimus provides long-term efficiency results comparable with a tacrolimus-based regimen, which represents a common treatment standard after kidney transplantation. Both are superior to other investigated immunosuppressive regimens.


Ankyloglossia superior syndrome: Case report and updated literature review.
Ankyloglossia superior (palatoglossal adhesion) is an extremely rare congenital condition with only 14 previously reported cases. When found in conjunction with other congenital abnormalities, such as cleft palate, gastrointestinal malformations, and limb malformations, this anomaly is considered part of ankyloglossia superior syndrome. We present a case of a newborn female found to have ankyloglossia superior syndrome. Surgical repair is also described. We review the available literature and discuss theories regarding the etiology of ankyloglossia superior syndrome. Clinicians should have a high degree of suspicion for other congenital anomalies when a neonate is found to have ankyloglossia superior.


LSD and tryptamine effects on sleep/wakefulness and electrocorticogram patterns in intact cats.
Effects of intravenous infusions of LSD (3.75, 7.5, 15 microgram/kg over 5 min; crossover N = 4) and tryptamine (0.04, 0.08, 0.12 mg/kg/min for 150 min; crossover N = 6) were compared to saline in intact cats through observation of five sleep/waking patterns. Electrocorticogram (ECoG) was analyzed for frequency band indices and mean amplitude and frequency. LSD increases wakefulness and drowsiness and decreases spindle sleep and rapid eye movement (REM) sleep during the first 75 min (period 1). The increase in active wakefulness and decrease in REM sleep persist during period 2, with an increase in spindle sleep thereafter. LSD increases delta index and ECoG amplitude, with a decrease in ECoG frequency; these effects peaked in period 2. Tryptamine increases wakefulness and drowsiness during period 1, with decreases in spindle sleep and REM sleep. The increase in quiet wakefulness and decrease in REM sleep persist during period 2, but no significant tryptamine effect is seen in sleep/waking patterns after infusion ceases. ECoG frequency increases during tryptamine infusion (periods 1 and 2), while ECoG amplitude increases during periods 2 and 3. Thus LSD and tryptamine both increase wakefulness, decrease spindle sleep, and decrease REM sleep.


The endothelial cell protein C receptor: its role in thrombosis.
The protein C anticoagulant pathway plays a crucial role as a regulator of the blood clotting cascade. Protein C is activated on the vascular endothelial cell membrane by the thrombin-thrombomodulin complex. The endothelial protein C receptor binds protein C and further enhances protein C activation. Once formed, activated protein C down-regulates thrombin formation by inactivating factors Va and VIIIa and exerts cytoprotective effects through endothelial protein C receptor binding. An adequate generation of activated protein C depends on the precise assembly, on the surface of the endothelial cells, of thrombin, thrombomodulin, protein C, and endothelial protein C receptor. Therefore, any change in the efficiency of this assembly may cause a reduction or increase in activated protein C generation and modulate the risk of thrombosis. This review highlights the role of the endothelial protein C receptor in disease and discusses the association of its mutations with the risk of thrombosis.


Effects of modification of the transcription initiation site context on citrus tristeza virus subgenomic RNA synthesis.
Citrus tristeza virus (CTV), a member of the Closteroviridae, has a positive-sense RNA genome of about 20 kb organized into 12 open reading frames (ORFs). The last 10 ORFs are expressed through 3'-coterminal subgenomic RNAs (sgRNAs) regulated in both amounts and timing. Additionally, relatively large amounts of complementary sgRNAs are produced. We have been unable to determine whether these sgRNAs are produced by internal promotion from the full-length template minus strand or by transcription from the minus-stranded sgRNAs. Understanding the regulation of 10 sgRNAs is a conceptual challenge. In analyzing commonalities of a replicase complex in producing so many sgRNAs, we examined initiating nucleotides of the sgRNAs. We mapped the 5' termini of intermediate- (CP and p13) and low- (p18) produced sgRNAs that, like the two highly abundant sgRNAs (p20 and p23) previously mapped, all initiate with an adenylate. We then examined modifications of the initiation site, which has been shown to be useful in defining mechanisms of sgRNA synthesis. Surprisingly, mutation of the initiating nucleotide of the CTV sgRNAs did not prevent sgRNA accumulation. Based on our results, the CTV replication complex appears to initiate sgRNA synthesis with purines, preferably with adenylates, and is able to initiate synthesis using a nucleotide a few positions 5' or 3' of the native initiation nucleotide. Furthermore, the context of the initiation site appears to be a regulatory mechanism for levels of sgRNA production. These data do not support either of the established mechanisms for synthesis of sgRNAs, suggesting that CTV sgRNA production utilizes a different mechanism.


Performance objectives for the self-management of cystic fibrosis.
Performance objectives for the self-management of cystic fibrosis (CF) were developed and subjected to a two-stage content validation. A multidisciplinary team of health care professionals generated a list of 149 medical and adjustment performance objectives. Behaviors included monitoring symptoms and judging their significance, treating symptoms and communicating with health care providers about symptoms and treatment plans. In the first stage a panel of experts in the medical and behavioral aspects of CF rated each behavior. In general, the eleven panelists rated the 149 behaviors as somewhat important or important (mean 2.6, S.D. 0.17, on a 3-point scale). In the second stage, 84 of 155 CF center directors rated all behaviors as somewhat important or important (mean 2.9, S.D. 0.23). Specific behaviors related to medical regimens were more consistently rated as important than were those related to psychosocial adjustment. The performance objectives provide a framework for developing and evaluating health education programs for the self-management of CF in order to promote optimum health and adjustment.


Formulae describing frequency selectivity as a function of frequency and level, and their use in calculating excitation patterns.
The auditory filter may be considered as a weighting function representing frequency selectivity at a particular centre frequency. Its shape can be derived using the power-spectrum model of masking which assumes: (1) in detecting a signal in a masker the observer uses the single auditory filter giving the highest signal-to-masker ratio; (2) threshold corresponds to a fixed signal-to-masker ratio at the output of that filter. Factors influencing the choice of a masker to measure the auditory filter shape are discussed. Narrow-band maskers are unsuitable for this purpose, since they violate the assumptions of the power-spectrum model. A method using a notched-noise masker is recommended, and typical results using that method are presented. The variation of the auditory filter shape with centre frequency and with level, and the relationship of the auditory filter shape and the excitation pattern are described. A method of calculating the excitation pattern of any sound as a function of level is presented, and examples and applications are given. The appendix gives a Fortran program for calculating excitation patterns.


Peptidyl-puromycin synthesis by free and membrane-bound ribosomes.
The peptidyl transferase reaction, as measured by the formation of peptidyl-puromycin, was compared for free ribosomes and ribosomes bound to two types of membrane, the endoplasmic reticulum and the outer nuclear membrane. In most respects the reaction catalyzed by the three types of ribosome was similar, demonstrating that interaction of the 60 S ribosomal subunit with the membrane has little effect on the functioning of peptidyl transferase, a 60 S protein. However, both the rate and extent of synthesis of peptidyl puromycin were lower for ribosomes bound to the nuclear membrane than for free or microsome-bound ribosomes. This difference appears to be a direct consequence of the ribosome-membrane interaction, since ribosomes stripped from the nuclear membrane could not be distinguished from the other classes of ribosome.


Xylanase production by Aspergillus awamori in solid-state fermentation and influence of different nitrogen sources.
The use of purified xylan as a substrate for bioconversion into xylanases increases the cost of enzyme production. Consequently, there have been attempts to develop a bioprocess to produce such enzymes using different lignocellulosic residues. Filamentous fungi have been widely used to produce hydrolytic enzymes for industrial applications, including xylanases, whose levels in fungi are generally much higher than those in yeast and bacteria. Considering the industrial importance of xylanases, the present study evaluated the use of milled sugarcane bagasse, without any pretreatment, as a carbon source. Also, the effect of different nitrogen sources and the C:N ratio on xylanase production by Aspergillus awamori were investigated, in experiments carried out in solid-state fermentation. High extracellular xylanolytic activity was observed on cultivation of A. awamori on milled sugarcane bagasse and organic nitrogen sources (45 IU/mL for endoxylanase and 3.5 IU/mL for beta-xylosidase). Endoxylanase and beta-xylosidase activities were higher when sodium nitrate was used as the nitrogen source, when compared with peptone, urea, and ammonium sulfate at the optimized C:N ratio of 10:1. The use of yeast extract as a supplement to the these nitrogen sources resulted in considerable improvement in the production of xylanases, showing the importance of this organic nitrogen source on A. awamori metabolism.


Enhancement of the in vivo antibody response by an 8-derivatized guanine nucleoside.
The capacity of the C8-substituted guanine ribonucleosides to enhance the in vivo humoral immune response to the protein antigen, human gamma globulin (HGG), in A/J mice was evaluated. It has been shown recently that the C8-substituted guanine ribonucleosides are a new class of potent adjuvant for humoral immune responses to the sheep erythrocyte antigen. The current studies extend these findings to HGG with 8-bromoguanosine (8BrGuo), a representative of this group of nucleosides. The adjuvant activity of 8BrGuo in this system is highly dose and time dependent. Although 8BrGuo enhanced responses when injected either early (Day 0) or late (Day 4 or 5) after immunization, its administration on Day 1 or 2 most often led to no enhancement, suggesting that 8BrGuo may act on two events separated by a resistant stage in an ongoing immune response. The plaque-forming cell (PFC) response to HGG was enhanced optimally at doses as low as 1 mg 8BrGuo/mouse administered either on the day of immunization or 4 days thereafter. In contrast, however, serum anti-HGG antibody concentration assayed by enzyme-linked immunosorbent assay (ELISA) was enhanced only at doses of 10 mg or more, injected on the day of immunization, but doses as low as 1 mg were effective on Day 4. 8BrGuo was also an effective adjuvant when injected after antigen administration in incomplete Freund's adjuvant or when administered by several different routes (intraperitoneal, subcutaneous, oral).


Metal silicide/poly-Si Schottky diodes for uncooled microbolometers.
: Nickel silicide Schottky diodes formed on polycrystalline Si 〈P〉 films are proposed as temperature sensors of monolithic uncooled microbolometer infrared focal plane arrays. The structure and composition of nickel silicide/polycrystalline silicon films synthesized in a low-temperature process are examined by means of transmission electron microscopy. The Ni silicide is identified as a multi-phase compound composed of 20% to 40% of Ni3Si, 30% to 60% of Ni2Si, and 10% to 30% of NiSi with probable minor content of NiSi2 at the silicide/poly-Si interface. Rectification ratios of the Schottky diodes vary from about 100 to about 20 for the temperature increasing from 22â"ƒ to 70â"ƒ; they exceed 1,000 at 80 K. A barrier of around 0.95 eV is found to control the photovoltage spectra at room temperature. A set of barriers is observed in photo-electromotive force spectra at 80 K and attributed to the Ni silicide/poly-Si interface. Absolute values of temperature coefficients of voltage and current are found to vary from 0.3%â"ƒ to 0.6%/â"ƒ for forward bias and around 2.5%/â"ƒ for reverse bias of the diodes.


Characterization of a pathway-specific activator of edeine biosynthesis and improved edeine production by its overexpression in Brevibacillus brevis.
Edeines are a group of non-ribosomal antibacterial peptides produced by Brevibacillus brevis. Due to the significant antibacterial properties of edeines, increasing edeine yield is of great interest in biomedical research. Herein, we identified that EdeB, a member of the ParB protein family, significantly improved edeine production in B. brevis. First, overexpression of edeB in B. brevis X23 increased edeine production by 92.27%. Second, in vitro bacteriostasis experiment showed that edeB-deletion mutant exhibited less antibacterial activity. Third, RT-qPCR assay demonstrated that the expression of edeA, edeQ, and edeK, which are key components of the edeine biosynthesis pathway, in edeB-deletion mutant X23(ΔedeB) was significantly lower than that in wild-type B. brevis strain X23. Finally, electrophoretic mobility shift assay (EMSA) showed that EdeB directly bound to the promoter region of the edeine biosynthetic gene cluster (ede BGC), suggesting that EdeB improves edeine production through interaction with ede BGC in B. brevis.


Graphene-spindle shaped TiO₂ mesocrystal composites: facile synthesis and enhanced visible light photocatalytic performance.
Graphene (GR)-TiO2 mesocrystal composites were prepared by a facile template-free process based on the combination of sol-gel and solvothermal methods, and were characterized using field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), Raman spectroscopy, UV-vis diffuse reflectance spectroscopy (UV-vis DRS), nitrogen absorption and electron spin resonance (ESR). Visible light photocatalytic performance of GR-TiO2 composites was evaluated for photocatalytic degradation of organic dye Rhodamine B. It was found that the amount of graphene oxide (GO) added obviously affects morphologies of TiO2 mesocrystals and photocatalytic activities of as-prepared nanocomposites. Composites prepared in the presence of different amounts of GO all exhibit higher photocatalytic activity than pure TiO2 mesocrystals and P25, the composite obtained by using 20mg GO presents the most uniform TiO2 mesocrystals in the composite and shows the highest photocatalytic efficiency. The mechanism for the generation of TiO2 mesocrystals in the GR-TiO2 composite is proposed and possible reasons for the enhancement in visible light photocatalytic efficiency are also discussed.