[Drugs in cardiopulmonary resuscitation. (I) Administration].
After their classification and limitation based on their therapeutic measures to their most scientifically contrasted efficiency measures, this article reviews what is written for each pharmaceutical in the recommendations bearing more weight in our field, those of the American Heart Association and those of the European Resuscitation Council. A study is made of the various techniques which can be used in administering these drugs as well as their utility and suitability in the field of resuscitation.


Reversal of TGF-β1 stimulation of α-smooth muscle actin and extracellular matrix components by cyclic AMP in Dupuytren's-derived fibroblasts.
Myofibroblasts, a derived subset of fibroblasts especially important in scar formation and wound contraction, have been found at elevated levels in affected Dupuytren's tissues. Transformation of fibroblasts to myofibroblasts is characterized by expression of alpha- smooth muscle actin (α-SMA) and increased production of extracellular matrix (ECM) components, both events of relevance to connective tissue remodeling. We propose that increasing the activation of the cyclic AMP (cAMP)/protein kinase A signaling pathway will inhibit transforming growth factor-beta1 (TGF-β1)-induced ECM synthesis and myofibroblast formation and may provide a means to blunt fibrosis. Fibroblasts derived from areas of Dupuytren's contracture cord (DC), from adjacent and phenotypically normal palmar fascia (PF), and from palmar fascia from patients undergoing carpal tunnel release (CTR; CT) were treated with TGF-β1 (2 ng/ml) and/or forskolin (10 μM) (a known stimulator of cAMP). Total RNA and protein extracted was subjected to real time RT-PCR and Western blot analysis. The basal mRNA expression levels of fibronectin- extra domain A (FN1-EDA), type I (COL1A2) and type III collagen (COL3A1), and connective tissue growth factor (CTGF) were all significantly increased in DC- and in PF-derived cells compared to CT-derived fibroblasts. The TGF-β1 stimulation of α-SMA, CTGF, COL1A2 and COL3A1 was greatly inhibited by concomitant treatment with forskolin, especially in DC-derived cells. In contrast, TGF-β1 stimulation of FN1-EDA showed similar levels of reduction with the addition of forskolin in all three cell types. In sum, increasing cAMP levels show potential to inhibit the formation of myofibroblasts and accumulation of ECM components. Molecular agents that increase cAMP may therefore prove useful in mitigating DC progression or recurrence.


[Properties of rhythmically active neurons of lymphatic heart center of the frog Rana temporaria].
During extracellular recordings of the activity of spinal neuronal units in situ, it was found that rhythmically active neurons respond to antidromic stimulation of ventral roots (X, XI), which include motor heart fibers. The amplitude of the responses of these motoneurons to continuous rhythmic antidromic stimulation exhibits slow rhythmic oscillations, reaching the lowest value to the onset of the spontaneous spike potentials. In isolated spinal cord preparation (from VIII segment), rhythmic activity of the center studied was preserved, which indicates the capacity of central structures to automatic activity.


Estimating Left Ventricular Filling Pressure by Echocardiography.
The diagnosis of heart failure may be challenging because symptoms are rather nonspecific. Elevated left ventricular (LV) filling pressure may be used to confirm the diagnosis, but cardiac catheterization is often not practical. Echocardiographic indexes are therefore used as markers of filling pressure. This study investigated the feasibility and accuracy of comprehensive echocardiography in identifying patients with elevated LV filling pressure. We conducted a multicenter study of 450 patients with a wide spectrum of cardiac diseases referred for cardiac catheterization. Left atrial volume index, in combination with flow velocities and tissue Doppler velocities, was used to estimate LV filling pressure. Invasively measured pressure was used as the gold standard. Mean left ventricular ejection fraction (LVEF) was 47%, with 209 patients having an LVEF <50%. Invasive measurements showed elevated LV filling pressure in 58% of patients. Clinical assessment had an accuracy of 72% in identifying patients with elevated filling pressure, whereas echocardiography had an accuracy of 87% (p < 0.001 vs. clinical assessment). The combination of clinical and echocardiographic assessment was incremental, with a net reclassification improvement of 1.5 versus clinical assessment (p < 0.001). Echocardiographic assessment of LV filling pressure is feasible and accurate. When combined with clinical data, it leads to a more accurate diagnosis, regardless of LVEF.


Changes observed in the growth fraction, labeling index, duration of S phase, and total cell cycle times of HL-60 cells as they undergo differentiation in response to retinoic acid.
To assess the changes in the proliferation characteristics that occur during maturation, HL-60 cells were induced to differentiate along the granulocytic pathway by retinoic acid. Differentiation was documented by morphology, functional markers, and cytochemical staining. Durations of S phase, total cell cycle time, and the percentage of S-phase cells were determined simultaneously at each time point. In addition, the expression of two cell cycle related proteins with molecular weights of 110,000 (p110 measured by monoclonal antibody 5C2) and 145,000 (measured by monoclonal antibody p145) were measured to estimate the number of cycling cells or the "growth fraction." Our data demonstrate that as HL-60 cells undergo maturation in response to retinoic acid, a large proportion of cells exit from the cycle, the majority lose their proliferative potential, and the total cell cycle time becomes markedly longer. The slowing of the cell cycle seems to be the result of a prolongation in both S phase and the G1 phase of the cycle. We conclude that more mature myeloid cells cycle more slowly than immature cells. The clinical implications of these findings in myeloid leukemias are discussed.


[Effect of "Naftusia" water on some indicators of metabolism in rats].
It is shown that giving rats natural water "Naftusya" to drink during 5 days initiated a complex of variations in change of substances, conditioned by general adaptational reaction. Ultraviolet radiation of water has no effect on its balneoactivity while ultrafiltration attenuates it a little.


The role of plasma membrane aquaporins in regulating the bundle sheath-mesophyll continuum and leaf hydraulics.
Our understanding of the cellular role of aquaporins (AQPs) in the regulation of whole-plant hydraulics, in general, and extravascular, radial hydraulic conductance in leaves (K(leaf)), in particular, is still fairly limited. We hypothesized that the AQPs of the vascular bundle sheath (BS) cells regulate K(leaf). To examine this hypothesis, AQP genes were silenced using artificial microRNAs that were expressed constitutively or specifically targeted to the BS. MicroRNA sequences were designed to target all five AQP genes from the PLASMA MEMBRANE-INTRINSIC PROTEIN1 (PIP1) subfamily. Our results show that the constitutively silenced PIP1 (35S promoter) plants had decreased PIP1 transcript and protein levels and decreased mesophyll and BS osmotic water permeability (P(f)), mesophyll conductance of CO2, photosynthesis, K(leaf), transpiration, and shoot biomass. Plants in which the PIP1 subfamily was silenced only in the BS (SCARECROW:microRNA plants) exhibited decreased mesophyll and BS Pf and decreased K(leaf) but no decreases in the rest of the parameters listed above, with the net result of increased shoot biomass. We excluded the possibility of SCARECROW promoter activity in the mesophyll. Hence, the fact that SCARECROW:microRNA mesophyll exhibited reduced P(f), but not reduced mesophyll conductance of CO2, suggests that the BS-mesophyll hydraulic continuum acts as a feed-forward control signal. The role of AQPs in the hierarchy of the hydraulic signal pathway controlling leaf water status under normal and limited-water conditions is discussed.


Somatic genome variations in health and disease.
It is hard to imagine that all the cells of the human organism (about 10(14)) share identical genome. Moreover, the number of mitoses (about 10(16)) required for the organism's development and maturation during ontogeny suggests that at least a proportion of them could be abnormal leading, thereby, to large-scale genomic alterations in somatic cells. Experimental data do demonstrate such genomic variations to exist and to be involved in human development and interindividual genetic variability in health and disease. However, since current genomic technologies are mainly based on methods, which analyze genomes from a large pool of cells, intercellular or somatic genome variations are significantly less appreciated in modern bioscience. Here, a review of somatic genome variations occurring at all levels of genome organization (i.e. DNA sequence, subchromosomal and chromosomal) in health and disease is presented. Looking through the available literature, it was possible to show that the somatic cell genome is extremely variable. Additionally, being mainly associated with chromosome or genome instability (most commonly manifesting as aneuploidy), somatic genome variations are involved in pathogenesis of numerous human diseases. The latter mainly concerns diseases of the brain (i.e. autism, schizophrenia, Alzheimer's disease) and immune system (autoimmune diseases), chromosomal and some monogenic syndromes, cancers, infertility and prenatal mortality. Taking into account data on somatic genome variations and chromosome instability, it becomes possible to show that related processes can underlie non-malignant pathology such as (neuro)degeneration or other local tissue dysfunctions. Together, we suggest that detection and characterization of somatic genome behavior and variations can provide new opportunities for human genome research and genetics.


Characterization of monoclonal antibodies against the human immunodeficiency virus (HIV) gag products and their use in monitoring HIV isolate variation.
Monoclonal antibodies were raised against the gag proteins of a British isolate of the human immunodeficiency virus (HIV), CBL-1. Seven of the monoclonal antibodies recognized HIV gag-coded proteins of 55,000 mol. wt. (p55) and 24,000 mol. wt. (p24), three recognized p55/p18 and three p18 alone. Cross-competition assays suggested that at least two independent epitopes exist on the p24 cleaved from the p55 precursor, whereas a cluster of closely related epitopes was recognized by the p55/p18 and p18 antibodies. The panel of monoclonal antibodies was then used to compare by immunofluorescence the expression of gag-encoded antigenic determinants on HIV isolates from different geographical locations. Minor changes in epitope expression were observed in isolates from the U.S.A. and Haiti with the most notable changes being detected in isolates from Zaire, Tanzania and Uganda.


Forecasting emergency department overcrowding: A deep learning framework.
As the demand for medical cares has considerably expanded, the issue of managing patient flow in hospitals and especially in emergency departments (EDs) is certainly a key issue to be carefully mitigated. This can lead to overcrowding and the degradation of the quality of the provided medical services. Thus, the accurate modeling and forecasting of ED visits are critical for efficiently managing the overcrowding problems and enable appropriate optimization of the available resources. This paper proposed an effective method to forecast daily and hourly visits at an ED using Variational AutoEncoder (VAE) algorithm. Indeed, the VAE model as a deep learning-based model has gained special attention in features extraction and modeling due to its distribution-free assumptions and superior nonlinear approximation. Two types of forecasting were conducted: one- and multi-step-ahead forecasting. To the best of our knowledge, this is the first time that the VAE is investigated to improve forecasting of patient arrivals time-series data. Data sets from the pediatric emergency department at Lille regional hospital center, France, are employed to evaluate the forecasting performance of the introduced method. The VAE model was evaluated and compared with seven methods namely Recurrent Neural Network (RNN), Long short-term memory (LSTM), Bidirectional LSTM (BiLSTM), Convolutional LSTM Network (ConvLSTM), restricted Boltzmann machine (RBM), Gated recurrent units (GRUs), and convolutional neural network (CNN). The results clearly show the promising performance of these deep learning models in forecasting ED visits and emphasize the better performance of the VAE in comparison to the other models.


Combination Therapy of Etanercept and Fumarates versus Etanercept Monotherapy in Psoriasis: A Randomized Exploratory Study.
Biologics are a safe and efficacious therapy for psoriasis. The drug survival of biologics may be disappointing, primarily due to loss of efficacy. Therefore, safe combination treatments are sought to improve their clinical response. To assess the efficacy, safety and tolerability of the combination therapy of etanercept with fumarates versus etanercept monotherapy. Thirty-three patients with psoriasis were randomized 1:1 to receive etanercept combined with fumarates or etanercept monotherapy. The primary outcome measure was the difference in PASI-75 response after 24 weeks; additionally, a longitudinal analysis was performed. An important secondary outcome measure was the proportion of patients with a Physician Global Assessment (PGA) of clear or almost clear. Adverse events were collected throughout the study. In the combination therapy group, 78% (14 out of 18 patients) reached PASI-75 at week 24 versus 57% (8 out of 14 patients) in the monotherapy group (p = 0.27). The longitudinal analysis showed a PASI reduction of 5.97% per week for the combination therapy group and of 4.76% for the monotherapy group (p = 0.11). In the combination therapy group, 94% (17 out of 18 patients) of patients had a PGA of clear/almost clear versus 64% (9 out of 14 patients) in the monotherapy group (p = 0.064). The incidence of mild gastrointestinal complaints was higher in the combination group than in the monotherapy group. Using the PGA, combination therapy showed a trend towards faster improvement in the first 24 weeks. The difference in the PASI score between the two groups was not statistically significant. Addition of fumarates to etanercept for 48 weeks appeared safe with an acceptable tolerability.


Integrative Variation Analysis Reveals that a Complex Genotype May Specify Phenotype in Siblings with Syndromic Autism Spectrum Disorder.
It has been proposed that copy number variations (CNVs) are associated with increased risk of autism spectrum disorder (ASD) and, in conjunction with other genetic changes, contribute to the heterogeneity of ASD phenotypes. Array comparative genomic hybridization (aCGH) and exome sequencing, together with systems genetics and network analyses, are being used as tools for the study of complex disorders of unknown etiology, especially those characterized by significant genetic and phenotypic heterogeneity. Therefore, to characterize the complex genotype-phenotype relationship, we performed aCGH and sequenced the exomes of two affected siblings with ASD symptoms, dysmorphic features, and intellectual disability, searching for de novo CNVs, as well as for de novo and rare inherited point variations-single nucleotide variants (SNVs) or small insertions and deletions (indels)-with probable functional impacts. With aCGH, we identified, in both siblings, a duplication in the 4p16.3 region and a deletion at 8p23.3, inherited by a paternal balanced translocation, t(4, 8) (p16; p23). Exome variant analysis found a total of 316 variants, of which 102 were shared by both siblings, 128 were in the male sibling exome data, and 86 were in the female exome data. Our integrative network analysis showed that the siblings' shared translocation could explain their similar syndromic phenotype, including overgrowth, macrocephaly, and intellectual disability. However, exome data aggregate genes to those already connected from their translocation, which are important to the robustness of the network and contribute to the understanding of the broader spectrum of psychiatric symptoms. This study shows the importance of using an integrative approach to explore genotype-phenotype variability.


Indomethacin-induced renal insufficiency: recurrence on rechallenge.
We have reported a case of acute oliguric renal failure with hyperkalemia in a patient with cirrhosis, ascites, and cor pulmonale after indomethacin therapy. Prompt restoration of renal function followed drug withdrawal, while re-exposure to a single dose of indomethacin caused recurrence of acute reversible oliguria. Our case supports the hypothesis that endogenous renal prostaglandins play a role in the maintenance of renal blood flow when circulating plasma volume is diminished. Since nonsteroidal anti-inflammatory agents interfere with this compensatory mechanism and may cause acute renal failure, they should be used with caution in such patients.


Enhanced axonal metabolism during early natalizumab treatment in relapsing-remitting multiple sclerosis.
The considerable clinical effect of natalizumab in patients with relapsing-remitting multiple sclerosis might be explained by its possible beneficial effect on axonal functioning. In this longitudinal study, the effect of natalizumab on absolute concentrations of total N-acetylaspartate, a marker for neuronal integrity, and other brain metabolites is investigated in patients with relapsing-remitting multiple sclerosis by using MR spectroscopic imaging. In this explorative observational study, 25 patients with relapsing-remitting multiple sclerosis initiating natalizumab treatment were included and scanned every 6 months for 18 months. Additionally 18 matched patients with relapsing-remitting multiple sclerosis continuing treatment with interferon-β or glatiramer acetate were included along with 12 healthy controls. Imaging included short TE 2D-MR spectroscopic imaging with absolute metabolite quantification of total N-acetylaspartate, creatine and phosphocreatine, choline-containing compounds, myo-inositol, and glutamate. Concentrations were determined for lesional white matter, normal-appearing white matter, and gray matter. At baseline in both patient groups, lower concentrations of total N-acetylaspartate and creatine and phosphocreatine were found in lesional white matter compared with normal-appearing white matter and additionally lower glutamate in lesional white matter of patients receiving natalizumab. In those patients, a significant yearly metabolite increase was found for lesional white matter total N-acetylaspartate (7%, P < .001), creatine and phosphocreatine (6%, P = .042), and glutamate (10%, P = .028), while lesion volumes did not change. In patients receiving interferon-β/glatiramer acetate, no significant change was measured in lesional white matter for any metabolite, while whole-brain normalized lesion volumes increased. Patients treated with natalizumab showed an increase in total N-acetylaspartate, creatine and phosphocreatine, and glutamate in lesional white matter. These increasing metabolite concentrations might be a sign of enhanced axonal metabolism.


[Fulminant sepsis caused by Clostridium perfringens of urologic origin].
To present a case of fulminant sepsis caused by Clostridium perfringens of urological origin. An 81-year-old diabetic female (the only factor of immunodepression) presented complicated renal colic two days later with fulminant and fatal sepsis caused by Clostridium perfringens with signs of disseminated intravascular coagulation. The patient died one hour after the presentation of disseminated intravascular coagulation despite attempts to resuscitate the patient in the emergency department. Due to the fulminant course of the condition, it was not possible to demonstrate the presence of massive intravascular hemolysis characteristic of these conditions. Blood cultures obtained immediately after the patient died were positive for Clostridium perfringens. Sepsis is a possible complication of infection from Clostridium perfringens. It is more frequent in immune-depressed patients and carries a high mortality despite medical and surgical treatment. Although it is not the most frequent, the genitourinary tract is a known portal of entry that should not be forgotten as in the case described herein.


Laryngeal mask airway for surfactant administration in a newborn animal model.
Premature infants are subjected to adverse effects of intubation to benefit from surfactant. We hypothesized that administration of surfactant through a laryngeal mask airway (LMA) is as effective as administration through an endotracheal tube (ETT) and that time and physiologic changes during instrumentation will be less in the LMA group. This study is a randomized, controlled trial using newborn pigs. Lung injury was induced via surfactant washout. Animals were randomized into groups: 1) LMA placed, no surfactant administered (control; n = 8); 2) surfactant via an LMA (LMA group; n = 8); and 3) surfactant via an ETT (ETT group; n = 8). We demonstrated that partial pressure of oxygen in arterial blood (Pao2) levels of the LMA and ETT groups were not statistically different. Time for successful placement of LMA was 19 ± 1 s versus ETT 123 ± 35 s (mean ± SEM); number of attempts for successful LMA placement was 1.1 (1-2) versus ETT 1.9 (1-7) [mean (range)]. Administration of surfactant via an LMA compared with an ETT resulted in similar improvements in oxygenation. Placement of the device required less time and fewer attempts. These data suggest that further study in human neonates is justified. If proven effective, some infants with respiratory distress may be able to receive surfactant while avoiding intubation.


FTO Facilitates Lung Adenocarcinoma Cell Progression by Activating Cell Migration Through mRNA Demethylation.
The fat mass and obesity-associated protein (FTO) was identified as a critical demethylase involved in regulating cellular mRNA stability by removing N6-methyladenosine (m6A) residues from mRNA. Emerging evidence has revealed that FTO is deeply implicated in lung cancer. However, knowledge of the function of FTO in lung adenocarcinoma (LUAC) is limited. FTO and FTO R96Q (R96Q), an FTO missense mutant lacking demethylase activity, were ectopically overexpressed, and FTO was knocked down via siRNA in A549 and H1299 cells. The relationships between FTO with cell characteristics and mRNA m6A levels were explored. Furthermore, RNA sequencing was performed on A549 cells. FTO overexpression enhanced the proliferation, migration, and invasion ability of A549 and H1299 cells, decreased mRNA m6A levels. Interestingly, overexpression of R96Q, blunted the effects of FTO overexpression on cell proliferation and invasion. Through RNA sequencing analysis of A549 cells overexpressing FTO or R96Q and control A594 cells, 45 genes were identified as affected by m6A mRNA demethylation. Most of these genes were related to lung cancer, such as laminin γ2, thrombospondin 1, nerve growth factor inducible, integrin alpha11, and proprotein convertase subtilisin/kexin type 9. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that these genes are fundamental to cancer development processes, such as cell migration and extracellular matrix organization. Our research shows that FTO facilitates LUAC cell progression by activating cell migration through m6A demethylation; however, further research on the mechanism underlying FTO activity in LUAC is necessary.


Silencing of frataxin gene expression triggers p53-dependent apoptosis in human neuron-like cells.
Friedreich's ataxia (FRDA) is an autosomal recessive disease caused by mutations that produce a deficiency in frataxin. Despite the importance of neurodegeneration in FRDA, little is known about the consequences of frataxin deficiency in neuronal cells. Here we describe a neuronal cell model for FRDA based on the use of lentiviral vectors that carry minigenes encoding frataxin-specific shRNAs that silence the expression of this gene. These lentivectors can knockdown frataxin expression in human neuroblastoma SH-SY5Y cells, which results in large-scale cell death in differentiated neuron-like cells but not in undifferentiated neuroblastoma cells. Frataxin-deficient neuron-like cells appear to die through apoptosis that is accompanied by up-regulation of p53, PUMA and Bax and activation of caspase-3. No significant autophagy is observed in frataxin-deficient neuron-like cells and the pharmacological activation of autophagy does not significantly increase neuronal cell death in response to the frataxin deficiency. Cell death triggered by frataxin knockdown can be impaired by interference with p53, caspase inhibitors and gene transfer of FXN. These results suggest that frataxin gene silencing in human neuron-like cells may constitute a useful cell model to characterize the molecular changes triggered by frataxin deficiency in neurons, as well as to search for therapies that may protect against neurodegeneration.


Conjoint analysis versus rating and ranking for values elicitation and clarification in colorectal cancer screening.
To compare two techniques for eliciting and clarifying patient values for decision making about colorectal cancer (CRC) screening: choice-based conjoint analysis and a rating and ranking task. Using our decision lab registry and university e-mail lists, we recruited average risk adults ages 48-75 for a written, mailed survey. Eligible participants were given basic information about CRC screening and six attributes of CRC screening tests, then randomized to complete either a choice-based conjoint analysis with 16 discrete choice tasks or a rating and ranking task. The main outcome was the most important attribute, as determined from conjoint analysis or participant ranking. Conjoint analysis-based most important attribute was determined from individual patient-level utilities generated using multinomial logistic regression and hierarchical Bayesian modeling. Of the 114 eligible participants, 104 completed and returned questionnaires. Mean age was 57 (range 48-73), 70% were female, 88% were white, 71% were college graduates, and 62% were up to date with CRC screening. Ability to reduce CRC incidence and mortality was the most frequent most important attribute for both the conjoint analysis (56% of respondents) and rating/ranking (76% of respondents) groups, and these proportions differed significantly between groups (absolute difference 20%, 95% CI 3%, 37%, p =0.03). There were no significant differences between groups in proportion with clear values (p = 0.352), intent to be screened (p = 0.226) or unlabelled test preference (p = 0.521) Choice-based conjoint analysis produced somewhat different patterns of attribute importance than a rating and ranking task, but had little effect on other outcomes.


Development of a Me2SO-free cryopreservation medium and its long-term cryoprotection on the CAR-NK cells.
Cryopreservation is a crucial step in the supply process of off-the-shelf chimeric antigen receptor engineered natural killer (CAR-NK) cell products. Concerns have been raised over the clinical application of dimethyl sulfoxide (Me2SO) due to the potential for adverse reactions following infusion and limited cell-specific cytotoxic effects if misapplied. In this study, we developed a Me2SO-free cryopreservation medium specifically tailored for CAR-NK cells to address this limitation. The cryopreservation medium was formulated using human serum albumin (HSA) and glycerol as the base components. Following initial screening of seven clinically-compatible solutions, four with cryoprotective properties were identified. These were combined and optimized into a single formulation: IF-M. The viability, phenotype, and function of CAR-NK cells were evaluated after short-term and long-term cryopreservation to assess the effectiveness of IF-M, with Me2SO serving as the control group. The viability and recovery of CAR-NK cells in the IF-M group were significantly higher than those in the Me2SO group within 90 days of cryopreservation. Moreover, after 1 year of cryopreservation the cytotoxic capacity of CAR-NK cells cryopreserved with IF-M was comparable to that of fresh CAR-NK cells and significantly superior to that of CAR-NK cells cryopreserved in Me2SO. The CD107a expression intensity of CAR-NK cells in IF-M group was significantly higher than that of Me2SO group. No statistical differences were observed in other indicators under different cryopreservation times. These results underscore the robustness of IF-M as a suitable replacement for traditional Me2SO-based cryopreservation medium for the long-term cryopreservation and clinical application of off-the-shelf CAR-NK cells.


Selective high affinity retinoic acid receptor alpha or beta-gamma ligands.
Biological effects of retinoic acid (RA) are mediated through its binding to three closely related nuclear receptors (RAR alpha, RAR beta, and RAR gamma) belonging to the steroid-thyroid nuclear receptor family. RARs are able to modulate the transcription of specific genes by binding to responsive elements located in the promoter-enhancer region of these genes. As demonstrated by in situ hybridization, the distribution of each RAR type in the developing embryo, as well as in the adult, is not uniform. In this context, synthetic retinoids that would behave as selective ligands would be invaluable for studying the respective roles of each RAR type in cultured cells, whole animals, and embryos. Moreover, from a pharmacological point of view, such selective compounds may possess a higher therapeutic index and a lower teratogenic risk, because they might affect specific tissues and spare some others. As an approach to this problem, we have set up two complementary assays, (i) an in vitro binding assay to determine the Kd values of retinoids for RAR alpha, RAR beta, and RAR gamma and (ii) a functional assay in cultured cells to evaluate the potential of retinoids to transactivate, through their binding to one type of RAR, a reporter gene. The binding assay uses nuclear extracts of COS-7 cells transfected with vectors expressing RAR alpha, RAR beta, or RAR gamma. The functional assay is a measure of chloramphenicol acetyltransferase (CAT) activity in HeLa cells co-transfected with the expression vectors used in the binding assay and the reporter gene TRE-tk-CAT. Selective agonists for RAR alpha (Am80 and Am580) and RAR beta-RAR gamma (CD495 and CD564) were identified. However, compounds with pure RAR beta or RAR gamma selectivity have not yet been identified.


Indigenous and traditional plants: South African parents' knowledge, perceptions and uses and their children's sensory acceptance.
The dietary shift from indigenous and traditional plants (ITPs) to cash crops and exotic plant food sources increases the risk of malnutrition and other nutrition-related non-communicable diseases, especially in poor rural communities. Farm communities in South Africa have been associated with poor nutritional status and extreme poverty. ITPs have been found to be affordable sources of several micronutrients. However, knowledge of and the use of these plants are declining, and little is known about the child's acceptance of dishes prepared with ITPs. This knowledge can be used to improve the general acceptance of ITPs. This study aimed to gain insight into parents' knowledge and perceptions and their use of ITPs in a farming community in the North West Province and to assess children's acceptance of and preference for dishes made with African leafy vegetables (ALVs) and Swiss chard. Parents (n = 29) responsible for food preparation for children in grade 2 to 4 in two schools were purposively selected for four focus group discussions. A sensory evaluation assessed the children's (n = 98) acceptance of, preference for and intended consumption of dishes made with leafy vegetables. The dishes were made of Amaranthus spp., Cleome gynandra, Cucurbita maxima, Vigna unguiculata and Beta vulgaris. Parents mentioned 30 edible ITPs during the focus group discussions. Parents had knowledge of available ITPs and their use as food. Location, seasonal variation and rainfall affected the availability of and access to ITPs. Sun-dried ITPs were stored in sacks for later use. ITPs were perceived as healthy, affordable and delicious, hence acceptable to the parents. The children also evaluated the dishes made with ALVs as acceptable in terms of colour, smell and taste. Swiss chard was preferred, most likely because of the children's exposure to this vegetable. Children indicated that they would like to eat these leafy vegetables twice a week. These results look promising for the promotion of ITPs as a strategy to reduce malnutrition in rural farm communities and for potential inclusion of these micronutrient-rich ALVs in school feeding programmes to improve the nutritional status of children.


Intramedullary spinal ependymomas: analysis of a consecutive series of 82 adult cases with particular attention to patients with no preoperative neurological deficit.
Surgery should be considered for patients with intramedullary spinal ependymomas (ISE), particularly those presenting with a neurological deficit preoperatively. In contrast, it is still a debatable matter whether to recommend the same approach for patients with no neurological impairment. To investigate this matter, we analyzed the data of 82 consecutive patients with ISEs treated at our institution. We reviewed the medical charts of all ISE patients undergoing operation at our institution between 1985 and 2000. Particular attention was given to patients without neurological deficit before surgery. Eighty-two consecutive ISE patients were included in this study. Preoperatively, a neurological deficit of variable severity was present in 72 patients (Group A, 88%) and absent in 10 patients (Group B, 12%). In the latter group, seven patients had progressive and nonspecific pain as the only presenting symptom; two had arm dysesthesias; and in one patient, ISE was diagnosed incidentally. The mean duration of their symptoms was 21 months. We achieved a total tumor excision in nine patients and subtotal removal in one. At the last follow-up assessment (mean, 45 mo), all Group B patients remained at Grade I of the McCormick classification, except one, who deteriorated to Grade Ib. Furthermore, nonspecific pain diminished in three patients, stabilized in four, and worsened in one. Arm dysesthesias diminished in one patient and stabilized in the other. No surgery-related complication or recurrence was recorded in these patients. Surgery should be carefully considered for ISE patients with no objective neurological deficit preoperatively because, in our experience, it resolves their preoperative complaints in 30% of cases, stabilizes them in 60%, and worsens them in 10%.


Isolating the elusive islet.
The lack of a technique that allows mass isolation of intact, viable human islets is part of the reason that islet transplantation has not become available to the human diabetic. This report outlines the history of islet isolation and presents two new technical modifications that have been developed in the dog. Many of the current problems in islet isolation are presented, including the difficulty in obtaining enough human pancreatic tissue with minimal warm-ischemia time; inadequate distention of the pancreas to provide sufficient disruption for maximal enzymatic reaction to release intact islets; inefficient chopping methods; the use of collagenase of variable composition; different digestion methods for obtaining isolated islets; and inefficient methods for separating and purifying the islets from the ductal, acinar, and fibrous components. The first new modification involves distention of the dog pancreas through the venous system of the gland rather than the ductal system. This results in improved intralobular disruption, which improved the yield of isolated dog islets by permitting more efficient collagenase digestion. The second new modification eliminates the concept of isolating intact islets: the dog pancreas is digested by trypsin to a single-cell preparation that is partially purified by Ficoll gradients; further purification of the endocrine cells results from selective aggregation using rotational culture. This process produces pseudoislets that contain all the islet cell types and can be kept in culture for up to 4 wk, releasing their hormones in response to appropriate stimuli. These modifications may assist in the struggle to isolate the elusive human islet for safe and effective islet transplantation in the diabetic patient.


Psychometric properties of the SF-36 in the early post-stroke phase.
Patients who have had a stroke are a large group in whom long-term disability is common and therefore impaired general health is likely. The Short Form 36 (SF-36) is a popular measure of general health that has been used with this patient group, but not all aspects of its psychometric properties have been established for use in this context, and its use in the early post-stroke phase has been neglected. To examine the reliability, validity and sensitivity to change of the SF-36 (UK version I) in patients in the early post-stroke period. A prospective, observational study of stroke outcomes was carried out. From May 1996 to April 1997, patients who had had a stroke were identified by 24 general practices in Scotland and were recruited within 1 month of their stroke, whether in hospital or at home. Outcome measures including the SF-36 were administered at one, 3 and 6 months after onset. The internal consistency of the eight subscales at all three time-points was good except for 1 month Vitality (alpha = 0.6824) and 3 month General Health (alpha = 0.6650), which were borderline in comparison with the criterion value of 0.7. Construct validity was adequate overall, although correlations between Role Physical and General Health and the Barthel Index and Canadian Neurological Scale were lower than hypothesized. Most SF-36 subscales were sensitive to change between 1 and 3 months post-stroke, but none detected change between 3 and 6 months. There were some practical problems in using the SF-36 in an acutely unwell stroke population. However, analysis of psychometric properties suggested that most of the subscales were adequately reliable and valid. Sensitivity to change was poorer in the later stages of the study.


Comparative analyses of downstream signal transduction targets modulated after activation of the AT1 receptor by two β-arrestin-biased agonists.
G protein-coupled receptors (GPCRs) are involved in essentially all physiological processes in mammals. The classical GPCR signal transduction mechanism occurs by coupling to G protein, but it has recently been demonstrated that interaction with β-arrestins leads to activation of pathways that are independent of the G protein pathway. Also, it has been reported that some ligands can preferentially activate one of these signaling pathways; being therefore called biased agonists for G protein or β-arrestin pathways. The angiotensin II (AngII) AT1 receptor is a prototype GPCR in the study of biased agonism due to the existence of well-known β-arrestin-biased agonists, such as [Sar(1), Ile(4), Ile(8)]-AngII (SII), and [Sar(1), D-Ala(8)]-AngII (TRV027). The aim of this study was to comparatively analyze the two above mentioned β-arrestin-biased agonists on downstream phosphorylation events and gene expression profiles. Our data reveal that activation of AT1 receptor by each ligand led to a diversity of activation profiles that is far broader than that expected from a simple dichotomy between "G protein-dependent" and "β-arrestin-dependent" signaling. We observed clusters of activation profiles common to AngII, SII, and TRV027, as well as downstream effector activation that are unique to AngII, SII, or TRV027. Analyses of β-arrestin conformational changes after AT1 receptor stimulation with SII or TRV027 suggests that the observed differences could account, at least partially, for the diversity of modulated targets observed. Our data reveal that, although the categorization "G protein-dependent" vs. "β-arrestin-dependent" signaling can be of pharmacological relevance, broader analyses of signaling pathways and downstream targets are necessary to generate an accurate activation profile for a given ligand. This may bring relevant information for drug development, as it may allow more refined comparison of drugs with similar mechanism of action and effects, but with distinct side effects.


CpG-depleted adeno-associated virus vectors evade immune detection.
Due to their efficient transduction potential, adeno-associated virus (AAV) vectors are leading candidates for gene therapy in skeletal muscle diseases. However, immune responses toward the vector or transgene product have been observed in preclinical and clinical studies. TLR9 has been implicated in promoting AAV-directed immune responses, but vectors have not been developed to circumvent this barrier. To assess the requirement of TLR9 in promoting immunity toward AAV-associated antigens following skeletal muscle gene transfer in mice, we compared immunological responses in WT and Tlr9-deficient mice that received an AAV vector with an immunogenic capsid, AAVrh32.33. In Tlr9-deficient mice, IFN-γ T cell responses toward capsid and transgene antigen were suppressed, resulting in minimal cellular infiltrate and stable transgene expression in target muscles. These findings suggest that AAV-directed immune responses may be circumvented by depleting the ligand for TLR9 (CpG sequences) from the vector genome. Indeed, we found that CpG-depleted AAVrh32.33 vectors could establish persistent transgene expression, evade immunity, and minimize infiltration of effector cells. Thus, CpG-depleted AAV vectors could improve outcome of clinical trials of gene therapy for skeletal muscle disease.


Therapy of endocrine disease. Effects of chronic use of phosphodiesterase inhibitors on endothelial markers in type 2 diabetes mellitus: a meta-analysis.
Diabetes mellitus (DM) is associated with endothelial dysfunction, reducing nitric oxide-dependent vasodilation, and increasing production of pro-inflammatory factors, leading to an increased risk of long-term cardiovascular disease. As the effects of phosphodiesterase 5 inhibitors (PDE5i) on endothelial function have not been systematically investigated, we conducted a meta-analysis of available randomized clinical trials (RCTs). A thorough search of the literature was carried out. Relevant studies were considered according to RCT study design, enrollment of men with type 2 DM, chronic administration of PDE5i, and evaluation of endothelial function through both hemodynamic and endothelial inflammation-related parameters. Fifteen studies fulfilled the eligibility criteria but only six RCTs met the inclusion criteria and were analyzed for 476 diabetic men, 239 randomized to Sildenafil, and 237 to placebo respectively. Four RCTs evaluated flow-mediated dilation (FMD), demonstrating a weighted mean increase of 2.19% (95% CI 0.48 to 3.90). This result showed a high heterogeneity (I(2): 98%). Thus, a further sub-group meta-analysis was performed and this analysis confirmed a significant, Sildenafil-related FMD improvement. Sildenafil improved endothelin 1 and high sensitivity C-reactive protein by ∼-0.94 pg/ml and -0.36 mg/l, respectively, not reaching statistical significance (P=0.69 and P=0.22 respectively). Finally, Sildenafil administration significantly reduced serum levels of interleukin 6 (IL6, -0.82 pg/ml; 95% CI -1.58 to -0.07). This meta-analysis suggests a beneficial effect of chronic PDE5i administration on endothelial function. Chronic Sildenafil administration seems to improve hemodynamic (FMD) and serum pro-inflammatory makers (IL6) in diabetic men. Larger studies are needed to confirm the effects of chronic PDE5i on endothelial function.


Evaluation of the efficacy of bentonite from the south of Argentina to ameliorate the toxic effects of aflatoxin in broilers.
In vitro studies indicated that a sodium bentonite (SB) from southern Argentina had a high ability to sorb aflatoxin B1 (AFB1) from aqueous solution. We evaluated this compound for its ability to reduce the effects of total aflatoxins (AF; 5 mg AFB1/kg) in the diet of growing broiler chickens from 30 to 52 d of age. The diets were amended with 0.3% Argentinean SB to determine the effect of this compound during aflatoxicosis. When compared with the controls, BW gains were significantly (P < 0.05) lower for broilers fed diets containing AF alone (1,865 vs. 1,552 g). No differences were found between the BW gains of broiler chickens fed diets without AF (1,785 g) and those of chickens fed AF + SB (1,809 g). These results suggest that effects of AF treatment were ameliorated when SB was used in the broiler chick diets. The AF significantly (P < 0.05) decreased feed efficiency. Liver, kidney, and pancreas relative weights increased in chickens fed the diet containing AF alone. Alterations in the levels of serum total protein, albumin (ALB), and globulins (GLOB) were observed for AF diets, and moderate protection was provided by the sorbent. The ALB:GLOB ratio decreased in both groups of birds fed with the AF-contaminated diet, and we observed a moderate increase in this ratio by 0.3% addition of SB. The histopathological findings in liver sections of broiler fed diets with AF + SB indicated a nonprotective effect of this adsorbent, because a moderate hepatic steatosis was observed.


An intelligent videomonitoring system for fall detection at home: perceptions of elderly people.
We explored the perception and receptivity of elderly people regarding the introduction of an intelligent videomonitoring system (IVS) at home. Using a mixed methods design, 25 elderly people with a history of falls completed a structured interview (two questionnaires). In the year preceding the interview, 72% of the participants fell as many as seven times. Open-ended questions (qualitative data) were used to supplement the quantitative data. A content analysis (qualitative data) and a descriptive analysis (quantitative data) were carried out. The participants were 84% favourable or partially favourable to technologies which ensured home security and 96% favourable or partially favourable to the IVS. About half (48%) said that they would use it. The other participants did not wish to use it unless they had been left to live alone or if their health condition worsened. Thus the living conditions of the elderly appear to influence their perception and receptivity regarding the acceptance and use of an IVS.


Sexual reproduction and gene flow in the pine pathogen Dothistroma septosporum in British Columbia.
Dothistroma septosporum has caused a serious needle blight epidemic in the lodgepole pine forests in northwest British Columbia over the past several years. Although ascocarps had been observed in British Columbia, nothing was known about the contribution of sexual reproduction, gene flow and long-distance dispersal to the epidemic. Amplified fragment length polymorphism and mating-type markers in 19 sites were used to generate population and reproductive data. Overall, evidence suggests a mixed mode of reproduction. Haplotypic diversity was high, with 79 unique and 56 shared haplotypes (possible clones) identified from 192 fungal isolates. Overall, mating-type segregation did not differ significantly from 1:1; however, random mating was rejected in most populations in the index of association and parsimony tree-length permutation analyses using the full data set and, when using clone-corrected data sets, more of the smaller populations showed random mating. Two of the smaller populations consistently showed random mating for both tests using both clone-corrected and noncorrected data. High gene flow is suggested by no differentiation between 14 of the 19 sites, several of which came from young plantations where the pathogen was not likely present prior to the current outbreak. The remaining five sites showed some level of divergence, possibly due to historic separation and endemic pathogen populations. Results indicate a high evolutionary potential and long-distance dispersal in this pathogen, important to consider in future forest management.


Evolution of microscopic heterogeneity and dynamics in choline chloride-based deep eutectic solvents.
Deep eutectic solvents (DESs) are an emerging class of non-aqueous solvents that are potentially scalable, easy to prepare and functionalize for many applications ranging from biomass processing to energy storage technologies. Predictive understanding of the fundamental correlations between local structure and macroscopic properties is needed to exploit the large design space and tunability of DESs for specific applications. Here, we employ a range of computational and experimental techniques that span length-scales from molecular to macroscopic and timescales from picoseconds to seconds to study the evolution of structure and dynamics in model DESs, namely Glyceline and Ethaline, starting from the parent compounds. We show that systematic addition of choline chloride leads to microscopic heterogeneities that alter the primary structural relaxation in glycerol and ethylene glycol and result in new dynamic modes that are strongly correlated to the macroscopic properties of the DES formed.


A Randomized Controlled Trial on the Effect of Tapentadol and Morphine on Conditioned Pain Modulation in Healthy Volunteers.
Modulatory descending pathways, originating at supraspinal sites that converge at dorsal horn neurons, influence pain perception in humans. Defects in descending pain control are linked to chronic pain states and its restoration may be a valuable analgesic tool. Conditioned pain modulation (CPM) is a surrogate marker of descending inhibition that reduces the perception of pain from a primary test stimulus during application of a conditioning stimulus. Here the effects of the analgesics tapentadol, a combined mu-opioid receptor agonist and noradrenaline reuptake inhibitor, and morphine, a strong mu-opioid receptor agonist, were tested on CPM in a randomized, double-blind, placebo-controlled crossover trial in 12 healthy pain-free volunteers, to understand possible differences in mechanism of action between these opioids. On three occasions CPM responses were obtained 60-90 and 120-150 min following intake of tapentadol (100 mg immediate release tablet), morphine (40 mg immediate release tablet) or placebo. At both time points, CPM was detectable after treatment with placebo and tapentadol (peak pain ratings reduced by 20-30% after application of the conditioning stimulus) but not after morphine. Compared to placebo morphine displayed significantly less CPM: mean treatment difference 18.2% (95% CI 3.4 to 32.9%) at 60-90 min after drug intake and 19.5% (95% CI 5.7 to 33.2%) at 120-150 min after drug intake (p = 0.001). No difference in CPM between placebo and tapentadol was detected: mean treatment difference 1.5% (95% CI -11.6 to 14.6%) at 60-90 min after drug intake and 1.5% (95% CI -16.0 to 18.9%) at 120-150 min after drug intake (p = 0.60). Our data show that in volunteers morphine affects CPM, while tapentadol was without effect despite identical experimental conditions. These data confirm that tapentadol's main mechanism of action is distinct from that of morphine and likely related to the effect of adrenergic stimulation on descending controls. Netherlands Trial Register NTR2716.


Masked inverse folding with sequence transfer for protein representation learning.
Self-supervised pretraining on protein sequences has led to state-of-the art performance on protein function and fitness prediction. However, sequence-only methods ignore the rich information contained in experimental and predicted protein structures. Meanwhile, inverse folding methods reconstruct a protein's amino-acid sequence given its structure, but do not take advantage of sequences that do not have known structures. In this study, we train a masked inverse folding protein masked language model parameterized as a structured graph neural network. During pretraining, this model learns to reconstruct corrupted sequences conditioned on the backbone structure. We then show that using the outputs from a pretrained sequence-only protein masked language model as input to the inverse folding model further improves pretraining perplexity. We evaluate both of these models on downstream protein engineering tasks and analyze the effect of using information from experimental or predicted structures on performance.


Rate of tritium labeling of specific purines in relation to nucleic acid and particularly transfer RNA conformation.
The kinetics of the incorporation of tritium into the C-8 positions of purine units in nucleic acids has been studied. The polymers investigated include poly(A), poly(A): poly (U) duplex, a double-stranded viral RNA, tRNA, and DNA. In the random coil state, the kinetics of incorporation of tritium into the purine sites of the polymers are identical with those for the corresponding purine mononucleotides. When the nucleic acids are in their native conformations, however, the purine labeling rates are reduced below that expected for the free mononucleotides. The magnitude of the effect is remarkably dependent upon the particular nucleic acid. For example, at 37 degrees C the purines in double-stranded DNA label at a rate two- to threefold slower than the corresponding mononucleotides, but in a double-stranded viral RNA, a 30- to 40-fold effect is found. The data suggest a strong influence of microscopic helix structure on the rate of tritium incorporation. First-order rate constants for the exchange of tritium into specific purine sites in yeast tRNAPhe were also determined. This was done by partially labeling the nucleic acid in tritiated water, and subsequently removing free and loosely bound tritium. Under conditions where exchange-out does not occur, the nucleic acid was digested with specific nucleases; chromatographic separation then enabled specific activities of purines from specific sites to be obtained. The rate constants for these sites show a large variation. They are markedly reduced for those residues occurring in cloverleaf helical sections and, in certain cases, for those known from crystallographic data to be involved in tertiary interactions. As examples of bases that can participate in tertiary interactions, the crystal structures show A14 and G15 in special base-pairing arrangements. Both purines (A14 and G15) occur in single-stranded sections of the cloverleaf; both show markedly reduced C-8 hydrogen-exchange rates. On the other hand, rate constants for bases and regions known to be on the outside of the moleculesuch as the anticodon loop and the 3' terminusāre perturbed the least. In one instance, a base in the dihydrouridine loop believed to be involved in tertiary interactions, according to crystallographic studies, incorporates tritium as if it were relatively unperburbed by the tRNA structure. The structural interactions of this base may be partially or completely broken at 37 degrees C in solution.


Effects of thyroxine-driven precocious metamorphosis on maturation of adult-type allograft rejection responses in early thyroidectomized frogs.
In contrast to the adult pattern of allograft reactivity, larval South African clawed frogs (Xenopus) either become tolerant of adult major histocompatibility complex (MHC) disparate skin grafts or reject them slowly. Larvae fail to reject grafts that are incompatible at minor histocompatibility (H) loci (MHC-identical); rather, they become immunologically tolerant. We report here that early thyroidectomized (thyroidx) larvae, forced to metamorphose precociously on a regimen of thyroxine (T4) treatment, showed larva-like responses in that their ability to reject skin allografts that differed by minor H loci or one MHC haplotype was significantly impaired relative to age-matched intact postmetamorphic controls. During the normal ontogeny of Xenopus, lymphocyte numbers increase in larval life, decrease during metamorphosis, and increase to adult levels at 6-12 months of age. Thyroidectomy and a low-dose regimen of thyroxine treatment limited the number of lymphocytes that developed in larval organs and very dramatically reduced those populations below control levels during metamorphosis. Because these changes were stage-dependent rather than age-dependent, the second wave of lymphopoiesis in thyroidx frogs treated with low T4 may occur out of synchrony with the expression of Class I MHC antigens. Impaired allograft rejection, therefore, may reflect the absence of a population of cells that can effectively recognize minor H alloantigens and/or self and allo Class I antigens.


Structural determinants in streptococcal unsaturated glucuronyl hydrolase for recognition of glycosaminoglycan sulfate groups.
Pathogenic Streptococcus agalactiae produces polysaccharide lyases and unsaturated glucuronyl hydrolase (UGL), which are prerequisite for complete degradation of mammalian extracellular matrices, including glycosaminoglycans such as chondroitin and hyaluronan. Unlike the Bacillus enzyme, streptococcal UGLs prefer sulfated glycosaminoglycans. Here, we show the loop flexibility for substrate binding and structural determinants for recognition of glycosaminoglycan sulfate groups in S. agalactiae UGL (SagUGL). UGL also degraded unsaturated heparin disaccharides; this indicates that the enzyme released unsaturated iduronic and glucuronic acids from substrates. We determined the crystal structures of SagUGL wild-type enzyme and both substrate-free and substrate-bound D175N mutants by x-ray crystallography and noted that the loop over the active cleft exhibits flexible motion for substrate binding. Several residues in the active cleft bind to the substrate, unsaturated chondroitin disaccharide with a sulfate group at the C-6 position of GalNAc residue. The sulfate group is hydrogen-bonded to Ser-365 and Ser-368 and close to Lys-370. As compared with wild-type enzyme, S365H, S368G, and K370I mutants exhibited higher Michaelis constants toward the substrate. The conversion of SagUGL to Bacillus sp. GL1 UGL-like enzyme via site-directed mutagenesis demonstrated that Ser-365 and Lys-370 are essential for direct binding and for electrostatic interaction, respectively, for recognition of the sulfate group by SagUGL. Molecular conversion was also achieved in SagUGL Arg-236 with an affinity for the sulfate group at the C-4 position of the GalNAc residue. These residues binding to sulfate groups are frequently conserved in pathogenic bacterial UGLs, suggesting that the motif "R-//-SXX(S)XK" (where the hyphen and slash marks in the motif indicate the presence of over 100 residues in the enzyme and parentheses indicate that Ser-368 makes little contribution to enzyme activity) is crucial for degradation of sulfated glycosaminoglycans.


Psoriatic arthritis and synovitis, acne, pustulosis, hyperostosis, and osteitis syndrome.
Psoriatic arthritis is an inflammatory arthropathy associated with psoriasis, and its clinical presentation varies from case to case. Distal interphalangeal involvement is characteristic but not seen in all patients. Enthesopathy, including that of the spine, is common and contributes to the classification of psoriatic arthritis as a seronegative spondyloarthropathy. The etiopathogenesis of psoriatic arthritis is not well understood, and evolution as measured by follow-up is variable. Treatment includes nonsteroidal anti-inflammatory drugs and some of the drugs used in the treatment of rheumatoid arthritis. Sulfasalazine and, in the more severe cases, cyclosporine, are being studied for efficacy and tolerance. Some cases of psoriatic arthritis are associated with an inflammatory bone disease, frequently seen on the anterior chest wall, which is part of the newly described SAPHO (synovitis, acne, pustulosis, hyperostosis, and osteitis) syndrome. SAPHO syndrome is characterized by this bone involvement, which can affect the spine and peripheral bones. Various skin conditions are associated with this syndrome, but they do not necessarily occur in all cases. Chronic recurrent multifocal osteomyelitis, which is seen mostly in children, may be a presentation of SAPHO syndrome. Associations with sacroiliitis, bowel disease, and psoriasis link SAPHO syndrome with the spondyloarthropathies.


Researchers' views on return of incidental genomic research results: qualitative and quantitative findings.
Comprehensive genomic analysis including exome and genome sequencing is increasingly being utilized in research studies, leading to the generation of incidental genetic findings. It is unclear how researchers plan to deal with incidental genetic findings. We conducted a survey of the practices and attitudes of 234 members of the US genetic research community and performed qualitative semistructured interviews with 28 genomic researchers to understand their views and experiences with incidental genetic research findings. We found that 12% of the researchers had returned incidental genetic findings, and an additional 28% planned to do so. A large majority of researchers (95%) believe that incidental findings for highly penetrant disorders with immediate medical implications should be offered to research participants. However, there was no consensus on returning incidental results for other conditions varying in penetrance and medical actionability. Researchers raised concerns that the return of incidental findings would impose significant burdens on research and could potentially have deleterious effects on research participants if not performed well. Researchers identified assistance needed to enable effective, accurate return of incidental findings. The majority of the researchers believe that research participants should have the option to receive at least some incidental genetic research results.


Functional mapping of androgen receptor enhancer activity.
Androgen receptor (AR) is critical to the initiation, growth, and progression of prostate cancer. Once activated, the AR binds to cis-regulatory enhancer elements on DNA that drive gene expression. Yet, there are 10-100× more binding sites than differentially expressed genes. It is unclear how or if these excess binding sites impact gene transcription. To characterize the regulatory logic of AR-mediated transcription, we generated a locus-specific map of enhancer activity by functionally testing all common clinical AR binding sites with Self-Transcribing Active Regulatory Regions sequencing (STARRseq). Only 7% of AR binding sites displayed androgen-dependent enhancer activity. Instead, the vast majority of AR binding sites were either inactive or constitutively active enhancers. These annotations strongly correlated with enhancer-associated features of both in vitro cell lines and clinical prostate cancer samples. Evaluating the effect of each enhancer class on transcription, we found that AR-regulated enhancers frequently interact with promoters and form central chromosomal loops that are required for transcription. Somatic mutations of these critical AR-regulated enhancers often impact enhancer activity. Using a functional map of AR enhancer activity, we demonstrated that AR-regulated enhancers act as a regulatory hub that increases interactions with other AR binding sites and gene promoters.


The measurement of state engagement in actions beyond basic professional duties.
The goal of this paper is to present the phenomenon of engagement in actions that go beyond basic professional duties, and the factor structure and other psychometric properties of the daily measurements of this construct, which corresponds with the growing interest of researchers in the phenomenon of employee engagement, extra-role behaviors and the factors that affect them on a daily basis. The research was carried out among 62 full-time or part-time working students of the humanities and social sciences (79% of whom were female), using a 5-day on-line daily survey. The main measure was a modified Polish version of the 9-item questionnaire consistent with the Utrecht Work Engagement Scale (UWES), which was prepared on the basis of the adaptation of the State Work Engagement (SWE) questionnaire proposed by Breevaart et al. (2012) to measure daily work engagement. The obtained results have shown that the adapted version of the SWE questionnaire is a reliable and valid measure which can be used to measure daily fluctuations in beyond-duties engagement. The 3-factor multilevel model has a satisfactory fit. The study contributes to the current body of literature by examining state engagement in actions that go beyond basic professional duties and the validity of its measurement. It also shows that giving employees more autonomy and feedback may positively influence their beyond-duties engagement. Med Pr. 2020;71(2):127-36.


The Alternatives to Traditional Materials for Subsoil Stabilization and Embankments.
Major infrastructure projects require significant amount of natural materials, often followed by the soft soil stabilization using hydraulic binders. This paper presents the results of a laboratory study of alternative waste materials (fly ash and slag) that can be used for earthworks. Results of high plasticity clay stabilization using fly ash from Serbian power plants are presented in the first part. In the second part of the paper, engineering properties of ash and ash-slag mixtures are discussed with the emphasis on the application in road subgrade and embankment construction. Physical and mechanical properties were determined via following laboratory tests: Specific gravity, grain size distribution, the moisture-density relationship (Proctor compaction test), unconfined compressive strength (UCS), oedometer and swell tests, direct shear and the California bearing ratio (CBR). The results indicate the positive effects of the clay stabilization using fly ash, in terms of increasing strength and stiffness and reducing expansivity. Fly ashes and ash-slag mixtures have also comparable mechanical properties with sands, which in combination with multiple other benefits (lower energy consumption and CO2 emission, saving of natural materials and smaller waste landfill areas), make them suitable fill materials for embankments, especially considering the necessity for sustainable development.


[Overexpression of connexin 40 (Cx40) inhibits the proliferation of H9c2 cardiomyocytes in rats by cell cycle arrest].
Objective To establish a stable strain of H9c2 cardiomyocytes overexpressing Cx40 and preliminarily investigate the effect of lentiviral vector-mediated Cx40 protein overexpression on the proliferation of H9c2 cells and its related mechanisms. Methods The Cx40 gene fragment was cloned from H9c2 cells by PCR and linked with lentivirus vector pLVX-IRES-Puro to obtain the recombinant plasmid pLVX-Flag-Cx40. Recombinant lentiviral particles carrying Flag-Cx40 were obtained by cotransfection with packaging plasmids into HEK293T cells. A stable expression strain (H9c2-Flag-Cx40 cell) was screened from infected H9c2 cells by purinomycin. The expression of Cx40 protein was detected by Western blot analysis, and the effect of Cx40 on H9c2 cells proliferation was determined by CCK-8 assay; cell cycle changes were measured by flow cytometry; the expression of the cell cycle protein cyclin D1 was detected by qRT-PCR and Western blot analysis. Co-immunoprecipitation (Co-IP) immunoprecipitation and Western blot analysis were used to identify the binding of Cx40 and Yes associated protein (YAP) in H9c2 cells; cytoplasmic and cytosolic proteins were isolated to detect the effect of Cx40 on the localization of YAP using Western blot analysis. Results Sequencing results showed that the recombinant pLVX-Flag-Cx40 expression vector was successfully established. A stable transfected cell line containing recombinant Flag-Cx40 lentivirus (H9c2-Flag-Cx40 cell) was successfully constructed from H9c2 cells. Compared with the control group, overexpression of Cx40 significantly reduced the proliferation of H9c2 cells, arrested the cell cycle at G0/G1 and reduced cyclin D1 expression. A significant increase in YAP expression was observed in the cytoplasm of the H9c2-Flag-Cx40 stable cell line, while the expression in the nucleus was significantly reduced. Cx40 bound to YAP in the cytoplasm and prevented it from entering the nucleus to play the role of transcriptional coactivation. Conclusion Overexpression of Cx40 induces cell-cycle arrest at G0/G1 phase and inhibits the proliferation in H9c2 cells.


Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic studies along the northern Colorado front range.
Epidemiologic evaluations were made of farm personnel on vesicular stomatitis-affected premises along the front range of the Rocky Mountains in Colorado during the 1982 epizootic. A similar antibody prevalence was noted to that of veterinarians and research and regulatory personnel who were involved with the same epizootic. Risk of infection resulted from intimate physical contact with infected horses or cows. Incidence and infection rates in horses were 45%; rates in cows were much lower, only 5%. Some epidemiologic clues were gained by a detailed study of an equine ranch. The pasture was incriminated as the area of highest risk, where 100% infection rates were noted. Horses in open pens and barns were at lower risk. Severe clinical disease in horses resulted in higher neutralizing antibody titers than inapparent or mild infection. Maternal antibody was detected in foals up to 4 months of age, and the level of antibody in the foal was a reflection of the dam's antibody level.


PLGA microspheres for the delivery of a novel subunit TB vaccine.
Biodegradable poly(dl-lactide-co-glycolide) microspheres were prepared using a modified double emulsion solvent evaporation method for the delivery of the subunit tuberculosis vaccine (Ag85B-ESAT-6), a fusion protein of the immunodominant antigens 6-kDa early secretory antigenic target (ESAT-6) and antigen 85B (Ag85B). Addition of the cationic lipid dimethyl dioctadecylammonium bromide (DDA) and the immunostimulatory trehalose 6,6'-dibehenate (TDB), either separately or in combination, was investigated for the effect on particle size and distribution, antigen entrapment efficiency, in vitro release profiles and in vivo performance. Optimised formulation parameters yielded microspheres within the desired sub-10 microm range (1.50 +/- 0.13 microm), whilst exhibiting a high antigen entrapment efficiency (95 +/- 1.2%) and prolonged release profiles. Although the microsphere formulations induced a cell-mediated immune response and raised specific antibodies after immunisation, this was inferior to the levels achieved with liposomes composed of the same adjuvants (DDA-TDB), demonstrating that liposomes are more effective vaccine delivery systems compared with microspheres.


[Surgical treatment for traumatic injury of the ascending aorta and aortic arch; report of a case].
We herein report a rare case of surgical treatment for blunt traumatic injury of the ascending aorta and aortic arch. A 60-year-old male was crushed by an arm of a forklift while working. He suffered from multiple thoracic traumas, including injury of the ascending aorta and aortic arch, multiple fractures of the ribs and bilateral hemopneumothorax. An emergency surgery for aortic repair was performed because there were no other severe hemorrhagic compilations due to organ injures. There was a massive hematoma around the aortic arch, and the intimae of several parts of the distal ascending aorta and aortic arch were disrupted circumferentially. The aorta was replaced with a prosthetic graft from sinotubular junction to the aortic arch, between the left common carotid artery and the left subclavian artery. The patient's postoperative course was uneventful, and he was transferred to a rehabilitation facility on the 13th day after surgery.


Short- and long-term effects of season-long infestation of Lycorma delicatula (Hemiptera: Fulgoridae) on young apple (Malus domestica) and peach (Prunus persica) trees.
Lycorma delicatula White, commonly called spotted lanternfly, is an invasive fulgorid that was first detected in the United States in Pennsylvania in 2014 and has spread across Eastern states. Lycorma delicatula is a phloem feeder, and feeding of persistent populations can cause wilt and dieback in host plants. Vineyards in infested regions have experienced vine loss, where damage has been attributed to L. delicatula. Nymphal and adult lifestages have been observed on cultivated apple and peach trees, but the feeding effects of this pest on these crops have not been quantified. Here, we caged young potted apple and peach trees and infested them with 50 nymphal or 25 adult L. delicatula per plant for 3 months and tracked plant health. Horticultural measurements including trunk diameter, height, leaf drop, and photosynthesis rates were recorded before, during, and after the infestation. To evaluate the long-term effects of infestation, all apple and peach trees were removed from pots and planted in an orchard block where their phenology, growth, and physiology could be evaluated the following growing season. Short- and long-term measurements showed no significant differences in apple tree health between infested and noninfested trees. There was a significant reduction in the growth of trunk diameter in peach trees during exposure to early-instar nymphs; infested peach trees also had significantly increased frost damage to buds the following spring. However, there were no long-term impacts on peach growth and physiology. These results indicate that L. delicatula likely will not be a major threat to these deciduous fruit crops.


Binding activity of 5alpha-reduced gestagens to the progestin receptor from African elephant (Loxodonta africana).
Recent findings in the African elephant (Loxodonta africana) indicate that the major progestins contained within and biosynthesized by corpora lutea are 5alpha-reduced metabolites and that progesterone is quantitatively of minor importance. The specific gestagenic action within the reproductive tract of elephants was determined by measurement of relative binding affinity of the respective progestins to the gestagen receptor extracted from elephant endometrium. The cytosol was incubated with 40 nmol/liter [3H]ORG-2058 and increasing concentrations of the tested progestin. Progesterone (P4), 5alpha-pregnane-3,20-dione (DHP), and 5alpha-pregnane-3alpha-ol-20-one (5alpha-P-3OH) were used. The competition for binding sites on the progestin receptor was shown by decreasing counts measured after extraction with scintillation fluid. The progestin concentration which induced a 50% reduction of measured counts was estimated (C50) and relative binding affinity of progesterone to other progestins was calculated (RBA = C50progestin/C50p4). The relative binding affinity of DHP to P4 at the gestagen receptor of elephant endometrium was equivalent. The other 5alpha-reduced progestin (5alpha-P-3OH) showed no competition to the [3H]ORG-2058 receptor binding. We conclude that the biological significance of P4 and DHP at the receptor level is very similar. The higher quantitative levels of DHP in corpus luteum and serum support the hypothesis that this progestin is the major gestagen in the elephants, whereas 5alpha-P-3OH is an inactive metabolite.


Activation of JNK1, RSK2, and MSK1 is involved in serine 112 phosphorylation of Bad by ultraviolet B radiation.
The Bcl-2 family member Bad is a pro-apoptotic protein, and phosphorylation of Bad by cytokines and growth factors promotes cell survival in many cell types. Induction of apoptosis by UV radiation is well documented. However, little is known about UV activation of cell survival pathways. Here, we demonstrate that UVB induces Bad phosphorylation at serine 112 in JNK1, RSK2, and MSK1-dependent pathways. Inhibition of mitogen-activated protein (MAP) kinases including ERKs, JNKs, and p38 kinase by the use of their respective dominant negative mutant or a specific inhibitor for MEK1 or p38 kinase, PD98059 or SB202190, resulted in abrogation of UVB-induced phosphorylation of Bad at serine 112. Incubation of active MAP kinase members with Bad protein showed serine 112 phosphorylation of Bad by JNK1 only. However, activated RSK2 and MSK1, downstream kinases of ERKs and p38 kinase, respectively, also phosphorylated Bad at serine 112 in vitro. Cells from a Coffin-Lowry syndrome patient (deficient in RSK2) or expressing an N-terminal or C-terminal kinase-dead mutant of MSK1 were defective for UVB-induced serine 112 phosphorylation of Bad. Furthermore, MAP kinase pathway-dependent serine 112 phosphorylation was shown to be required for dissociation of Bad from Bcl-X(L). These data illustrated that UVB-induced phosphorylation of Bad at serine 112 was mediated through MAP kinase signaling pathways in which JNK1, RSK2, and MSK1 served as direct mediators.


Analysis of CaM-kinase signaling in cells.
A change in intracellular free calcium is a common signaling mechanism that modulates a wide array of physiological processes in most cells. Responses to increased intracellular Ca(2+) are often mediated by the ubiquitous protein calmodulin (CaM) that upon binding Ca(2+) can interact with and alter the functionality of numerous proteins including a family of protein kinases referred to as CaM-kinases (CaMKs). Of particular interest are multifunctional CaMKs, such as CaMKI, CaMKII, CaMKIV and CaMKK, that can phosphorylate multiple downstream targets. This review will outline several protocols we have used to identify which members and/or isoforms of this CaMK family mediate specific cellular responses with a focus on studies in neurons. Many previous studies have relied on a single approach such as pharmacological inhibitors or transfected dominant-negative kinase constructs. Since each of these protocols has its limitations, that will be discussed, we emphasize the necessity to use multiple, independent approaches in mapping out cellular signaling pathways.


Ionized or Total Magnesium levels, what should we measure in critical ill patients?
Monitoring and measuring magnesium (Mg) values are essential to prevent the development of numerous complications in perioperative medicine and critically ill patients. Although previous studies suggest that measuring free ionized magnesium (iMg) is more useful for estimating Mg status, clinicians currently rely on measurement of total serum magnesium to determine if supplemental magnesium is needed. In this review, we analyzed the recent literature to decide whether it is better to measure ionized serum Mg or total serum Mg when assessing magnesium status, whether iMg predicts clinical outcome, and what are the difficulties in measuring serum iMg levels in intensive care patients and perioperative medicine.


Epidermal growth factor 61 A/G polymorphism and uterine cervical cancer.
Cervical cancer, the second most common malignancy in women worldwide, is almost invariably associated with infection by human papillomavirus (HPV). However, although many women are infected with high-risk types of HPV, only a subset of infected women will ever develop cervical cancer. Therefore, host genetic factor may play a role in cervical carcinogenesis. Alterations in epidermal growth factor receptor (EGFR) are common events in cervical cancer. Therefore, we hypothesized that a functional polymorphism in the 5' untranslated region of the epidermal growth factor (EGF) gene, a natural ligand of the EGFR, may play a role in the cervical carcinogenesis and tumor invasiveness. We assessed the possible association between EGF +61 A/G polymorphism and cervical cancer risk in a hospital-based case-control study among 337 Korean women (168 cases, 169 age-matched controls). The frequencies of EGF +61 allele and genotype were not different between cases and controls. We observed increasing trend of lymph node metastasis from A/A homozygous genotype toward G/G homozygous genotype. We did not find any evidence that EGF +61 A/G polymorphism was associated with individual susceptibility of cervical cancer. However, although it was not statistically significant, the increasing trend of lymph node metastasis according to EGF genotype suggests the possibility that individual variance of EGF expression may be associated with cervical cancer invasiveness. We also confirmed that there exists striking ethnic heterogeneity of EGF genotype between Caucasian and East Asian population.


Validating the genomic signature of pediatric septic shock.
We previously generated genome-wide expression data (microarray) from children with septic shock having the potential to lead the field into novel areas of investigation. Herein we seek to validate our data through a bioinformatic approach centered on a validation patient cohort. Forty-two children with a clinical diagnosis of septic shock and 15 normal controls served as the training data set, while 30 separate children with septic shock and 14 separate normal controls served as the test data set. Class prediction modeling using the training data set and the previously reported genome-wide expression signature of pediatric septic shock correctly identified 95-100% of controls and septic shock patients in the test data set, depending on the class prediction algorithm and the gene selection method. Subjecting the test data set to an identical filtering strategy as that used for the training data set, demonstrated 75% concordance between the two gene lists. Subjecting the test data set to a purely statistical filtering strategy, with highly stringent correction for multiple comparisons, demonstrated <50% concordance with the previous gene filtering strategy. However, functional analysis of this statistics-based gene list demonstrated similar functional annotations and signaling pathways as that seen in the training data set. In particular, we validated that pediatric septic shock is characterized by large-scale repression of genes related to zinc homeostasis and lymphocyte function. These data demonstrate that the previously reported genome-wide expression signature of pediatric septic shock is applicable to a validation cohort of patients.


Porphyromonas gingivalis virulence in mice: induction of immunity to bacterial components.
Selected cell envelope components of Porphyromonas gingivalis were tested in a BALB/c mouse model in an attempt to elucidate further the outer membrane components of this putative oral pathogen that might be considered as virulence factors in host tissue destruction. Lipopolysaccharide (LPS), outer membrane, and outer membrane vesicles of P. gingivalis W50, ATCC 53977, and ATCC 33277 were selected to examine an immunological approach for interference with progressing tissue destruction. Mice were actively immunized with heat-killed (H-K) or Formalin-killed (F-K) whole cells or with the outer membrane fraction, LPS, or outer membrane vesicles of the invasive strain P. gingivalis W50. The induction of invasive spreading lesions with tissue destruction and lethality were compared among different immunization groups in normal, dexamethasone-treated (dexamethasone alters neutrophil function at the inflammatory site), and galactosamine-sensitized (galactosamine sensitization increases endotoxin sensitivity) mice after challenge infection with the homologous strain (W50) and heterologous strains (ATCC 53977 and ATCC 33277). Enzyme-linked immunosorbent assay analyses revealed significantly elevated immunoglobulin G and M antibody responses after immunization with H-K or F-K cells or the outer membrane fraction compared with those of nonimmunized mice. The killed whole-cell vaccines provided significantly greater protection against challenge infection in normal mice (decreased lesion size and death) than did either the outer membrane fraction or LPS immunization. The lesion development observed in dexamethasone-pretreated mice was significantly enhanced compared with that of normal mice after challenge with P. gingivalis. Immunization with P. gingivalis W50 provided less protection against heterologous challenge infection with P. gingivalis ATCC 53977; however, some species-specific antigens were recognized and induced protective immunity. Only viable P. gingivalis induced a spreading lesion in normal, dexamethasone-treated, or galactosamine-sensitized mice; F-K or H-K bacteria did not induce lesions. The F-K and outer membrane vesicle immunization offered greater protection from lesion induction than did the H-K immunogen after challenge infection simultaneous with galactosamine sensitization. The H-K cell challenge with galactosamine sensitization produced 100% mortality without lesion induction, suggesting that LPS or LPS-associated outer membrane molecules were functioning like endotoxin. Likewise, P. gingivalis W50 LPS (1 micrograms per animal) administered intravenously produced 80% mortality in galactosamine-sensitized mice. In contrast to the effects of immunization on lesion development, immunization with H-K or F-K cells or LPS provided no protection against intravenous challenge with LPS; 100% of the mice died from acute endotoxin toxicity.(ABSTRACT TRUNCATED AT 400 WORDS)


Contrasting effects of steroids and mizoribine on macrophage activation and glomerular lesions in rat thy-1 mesangial proliferative glomerulonephritis.
Macrophages with a pro-inflammatory (M1) phenotype mediate renal injury in proliferative forms of glomerulonephritis, while alternatively activated (M2) macrophages are thought to be anti-inflammatory and promote repair. Glucocorticoids, the mainstay therapy for proliferative glomerulonephritis, can induce alternative macrophage activation in vitro, but it is unknown whether this occurs in vivo and if this is required for glucocorticoid responsiveness. In addition, clinical studies have suggested that the ability of mizoribine (MZR) to suppress steroid-resistant proliferative glomerulonephritis may operate via inhibiting pro-inflammatory macrophage activation. This study examined prednisolone (PSL) and/or MZR treatment of rat Thy-1 disease - a model in which macrophages promote mesangial proliferative glomerulonephritis. PSL treatment of Thy-1 nephritis induced an M2-like macrophage phenotype, but failed to modify mesangial hypercellularity and actually exacerbated global glomerulosclerosis. In contrast, MZR treatment reduced hypercellularity and glomerulosclerosis and suppressing both M1 and M2 markers of macrophage activation, with a selective reduction in CD169+ macrophages. Combined PSL/MZR treatment suppressed glomerular lesions and prevented steroid induction of an M2-like macrophage phenotype. In vitro, MZR prevented steroid induction of an M2 macrophage phenotype. Glucocorticoid induced alternative macrophage activation failed to ameliorate rat mesangial proliferative glomerulonephritis, whereas MZR suppression of this disease model was attributed, in part, to inhibition of M1-like pro-inflammatory macrophage activation.


[Evaluation of the medical orientation for the benzodiazepine side effects].
Benzodiazepines are among the most prescribed drugs indicated as anxiolytics, hypnotics, myorrelaxants or antiepileptics. Their main side effects are: reduced psychomotor activities, interaction to other drugs, like alcohol, and the development of dependence. In the present study it was evaluated the quality of the medical directions given to patients about those side effects. One hundred and twenty patients (39 men and 81 women), mean age of 48 years old, who visited pharmacies in Curitiba to buy prescribed benzodiazepines, were interviewed by an open or directive questionnaire designed to obtain the medical directions about benzodiazepines side effects. Directions about the three main benzodiazepines side effects were observed only in 13% of the patients, 27% had received at least two and 40% only one, while 19% reported no directions. It suggested that the medical orientation was precarious and its quality was not influenced by the patients' educational level, the kind of medical attendance or the specialty of the physician. The main orientation cited was "don't drink" (85%), followed by "don't drive or operate machines" (46%), while few orientations about the risk to develop dependence on benzodiazepines was observed (31%). This suggests that physicians were worried about the alcohol interaction risk, which can be dangerous for the patients. The high number of patients using benzodiazepines continuously for more than one year (61%), the unsuccessfully attempt to stop using BZD (94%) and the poor information about the duration of the treatment (22%) were all related to the low medical worry about dependence on benzodiazepines.


A Novel Spectrophotometric Method for Determination of Percarbonate by Using N, N-Diethyl-P-Phenylenediamine as an Indicator and Its Application in Activated Percarbonate Degradation of Ibuprofen.
Sodium percarbonate (SPC) concentration can be determined spectrophotometrically by using N, N-diethyl-p-phenylenediamine (DPD) as an indicator for the first time. The ultraviolet-visible spectrophotometry absorbance of DPD•+ measured at 551 nm was used to indicate SPC concentration. The method had good linearity (R2 = 0.9995) under the optimized experimental conditions (pH value = 3.50, DPD = 4 mM, Fe2+ = 0.5 mM, and t = 4 min) when the concentration of SPC was in the range of 0-50 μM. The blank spiked recovery of SPC was 95-105%. The detection limit and quantitative limit were 0.7-1.0 μM and 2.5-3.3 μM, respectively. The absorbance values of DPD•+ remained stable within 4-20 min. The method was tolerant to natural water matrix and low concentration of hydroxylamine (<0.8 mM). The reaction stoichiometric efficiency of SPC-based advanced oxidation processes in the degradation of ibuprofen was assessed by the utilization rate of SPC. The DPD and the wastewater from the reaction were non-toxic to Escherichia coli. Therefore, the novel Fe2+/SPC-DPD spectrophotometry proposed in this work can be used for accurate and safe measurement of SPC in water.


Intrauterine growth restriction does not alter response of protein synthesis to feeding in newborn pigs.
This study aimed to determine the effect of intrauterine growth restriction (IUGR) on the acute response of tissue protein synthesis to feeding in newborn pigs. Newborn pigs of sows fed either control or protein-restricted diets throughout gestation were designated C or IUGR, respectively. Both groups were either fasted for 9 h after birth or fed hourly 30 ml colostrum/kg body wt for 2.75 h after a 6-h fast. Fractional rates of tissue protein synthesis (Ks) were measured in vivo with a flooding dose of L-[4-3H]phenylalanine. Birth weight was reduced by 33% in IUGR pigs. IUGR had no effect on Ks in skeletal muscles, heart, liver, jejunum, or pancreas. Feeding stimulated tissue Ks similarly in C and IUGR pigs. Fasting plasma insulin concentrations and their rise with feeding were unaffected by IUGR. Plasma insulin-like growth factor I (IGF-I) concentrations were reduced by 42% in IUGR pigs and were not altered by feeding in either IUGR or C pigs. There were positive nonlinear relationships between tissue Ks and circulating concentrations of insulin. The results indicate that, in newborn pigs, tissue Ks are unaffected by IUGR, despite reduced plasma IGF-I concentrations. The efficiency with which nutrients stimulate tissue Ks is also not altered by IUGR, perhaps because the rise in plasma insulin concentrations with feeding is unaffected by IUGR.


Pipeline to Identify Hydroxyproline-Rich Glycoproteins.
Intrinsically disordered proteins (IDPs) are functional proteins that lack a well-defined three-dimensional structure. The study of IDPs is a rapidly growing area as the crucial biological functions of more of these proteins are uncovered. In plants, IDPs are implicated in plant stress responses, signaling, and regulatory processes. A superfamily of cell wall proteins, the hydroxyproline-rich glycoproteins (HRGPs), have characteristic features of IDPs. Their protein backbones are rich in the disordering amino acid proline, they contain repeated sequence motifs and extensive posttranslational modifications (glycosylation), and they have been implicated in many biological functions. HRGPs are evolutionarily ancient, having been isolated from the protein-rich walls of chlorophyte algae to the cellulose-rich walls of embryophytes. Examination of HRGPs in a range of plant species should provide valuable insights into how they have evolved. Commonly divided into the arabinogalactan proteins, extensins, and proline-rich proteins, in reality, a continuum of structures exists within this diverse and heterogenous superfamily. An inability to accurately classify HRGPs leads to inconsistent gene ontologies limiting the identification of HRGP classes in existing and emerging omics data sets. We present a novel and robust motif and amino acid bias (MAAB) bioinformatics pipeline to classify HRGPs into 23 descriptive subclasses. Validation of MAAB was achieved using available genomic resources and then applied to the 1000 Plants transcriptome project (www.onekp.com) data set. Significant improvement in the detection of HRGPs using multiple-k-mer transcriptome assembly methodology was observed. The MAAB pipeline is readily adaptable and can be modified to optimize the recovery of IDPs from other organisms.


The prevalence of suicidal ideation and suicidal attempts among Norwegian physicians. Results from a cross-sectional survey of a nationwide sample.
Physicians have a higher suicide rate than the general population or other academics. Little is known about the reasons for this. Analysing risk factors may be a valuable way of identifying reasons for the high suicide rate among physicians, thereby leading to preventive efforts. The present study is one of the first papers on suicidal thoughts and attempts among physicians. A questionnaire about suicidal thoughts (developed by E.S. Paykel) was completed by 1,063 of 1,476 active Norwegian physicians (72%). Lifetime prevalence ranged from 51.1% for feelings that life was not worth living to 1.6% for a suicide attempt. Risk factors were being female, living alone, and depression. Suicidal thoughts, however, were hardly attributed to working conditions. A high rate of suicide and a low rate of suicidal attempts support the hypothesis that physicians do not 'cry for help,' but are inclined to act out their suicidal impulses.


[The characteristics of the epithelium and endometrial extracellular matrix in miscarriage during early pregnancy associated with chronic endometritis].
In miscarriage associated with chronic endometritis, the uterine mucosa was found to have a lower expression of TGFR-beta, gelatinases and a higher exposure of TIMP-1 and EGFR, which confirms a pathogenetic association between the imbalance of growth factors, endoproteases and their inhibitors and points to the active involvement of these cytokines and endopeptidases in the inflammatory, reparative processes and remodeling of tissues. The imbalance of growth factors, proteases, and antiproteases is likely to be a cause of uterine mucosal remodeling with progressive collagenogenesis and atrophy of the glandular apparatus along with pathogen persistence, which may be one of the factors predisposing to miscarriage. This makes it necessary to eliminate the causative agent of the endometrial infectious-inflammatory process in the earliest periods of the onset of disease and permits the drawing up of approaches to targeted drug therapy for some pathogenetic links of disease.


Human exposure to allergenic pollens: a comparison between urban and rural areas.
Pollinosis is found more frequently in urban areas than in rural environments. This could be partly related to the different types of pollen exposure in these dissimilar areas. The objective of this study was to compare the distribution of pollen in these environments across an urbanization gradient. Daily pollen abundances were obtained in France using Hirst-type sensors. Sampling was conducted from January to June in 2003 and 2006 in a rural area, a semi-rural area and in two urban areas, which were characterized by several urbanization criteria. Total allergenic pollen abundance was higher in rural and semi-rural areas than in urban areas irrespective of the sampling year. Multivariate analyses showed that pollen exposures differed according to the type of area and were strongly explained by the urbanization gradient. Grass, ash, birch, alder, hornbeam, hazel and plantain pollen quantities exceeded the allergy threshold more often in rural settings than in urban areas. In urban areas, only plane pollen quantities exceeded the allergy threshold more often than in rural areas. Allergenic pollen exposure is higher in rural areas than in urban areas, and the most abundant pollen in each area did not originated from the same taxa. This result should be taken into account in epidemiological studies comparing allergies in rural and urban areas to adapt the panel of pollen extracts for human environmental exposure. In addition, this study highlights that some ornamental trees produce a large number of allergenic pollens and provide new sources of aeroallergens.


Numerical analysis of polarization splitter based on vertically coupled microring resonator.
The performance of a polarization splitter based on vertically coupled microring resonator is rigorously investigated by a combination of a 3D full vectorial film mode matching method with a 3D full vectorial coupled mode theory. The spectral responses of the structure for TE and TM mode are calculated, together with eigenmodes of uncoupled waveguides and scattering matrix of coupling region. The result shows that the response of microring resonator is indeed strongly polarization dependent and the resonance wavelengths are different for TE and TM mode. Such property allows for the design of wavelength-sensitive integrated polarization splitter. The influence of geometrical parameters on splitting ratio is investigated and the results indicate that the structure can have a splitting ratio greater than 20dB at 1.55 microm.