Tri- and tetra-substituted cyclen based lanthanide(III) ion complexes as ribonuclease mimics: a study into the effect of log Ka, hydration and hydrophobicity on phosphodiester hydrolysis of the RNA-model 2-hydroxypropyl-4-nitrophenyl phosphate (HPNP).
A series of tetra-substituted 'pseudo' dipeptide ligands of cyclen (1,4,7,10,-tetraazacyclododecane) and a tri-substituted 3'-pyridine ligand of cyclen, and the corresponding lanthanide(III) complexes were synthesised and characterised as metallo-ribonuclease mimics. All complexes were shown to promote hydrolysis of the phosphodiester bond of 2-hydroxypropyl-4-nitrophenyl phosphate (HPNP, τ1/2 = 5.87 × 10(3) h), a well known RNA mimic. The La(III) and Eu(III) tri-substituted 3'-pyridine lanthanide(III) complexes being the most efficient in promoting such hydrolysis at pH 7.4 and at 37 °C; with τ1/2 = 1.67 h for La(III) and 1.74 h for Eu(III). The series was developed to provide the opportunity to investigate the consequences of altering the lanthanide(III) ion, coordination ability and hydrophobicity of a metallo-cavity on the rate of hydrolysis using the model phosphodiester, HPNP, at 37 °C. To further provide information on the role that the log Ka of the metal bound water plays in phosphodiester hydrolysis the protonation constants and the metal ion stability constants of both a tri and tetra-substituted 3'pyridine complex were determined. Our results highlighted several key features for the design of lanthanide(III) ribonucelase mimics; the presence of two metal bound water molecules are vital for pH dependent rate constants for Eu(III) complexes, optimal pH activity approximating physiological pH (∼7.4) may be achieved if the log Ka values for both MLOH and ML(OH)2 species occur in this region, small changes to hydrophobicity within the metallo cavity influence the rate of hydrolysis greatly and an amide adjacent to the metal ion capable of forming hydrogen bonds with the substrate is required for achieving fast hydrolysis.


Mycobacterial nicotinate mononucleotide adenylyltransferase: structure, mechanism, and implications for drug discovery.
Nicotinate mononucleotide adenylyltransferase NadD is an essential enzyme in the biosynthesis of the NAD cofactor, which has been implicated as a target for developing new antimycobacterial therapies. Here we report the crystal structure of Mycobacterium tuberculosis NadD (MtNadD) at a resolution of 2.4 Å. A remarkable new feature of the MtNadD structure, compared with other members of this enzyme family, is a 310 helix that locks the active site in an over-closed conformation. As a result, MtNadD is rendered inactive as it is topologically incompatible with substrate binding and catalysis. Directed mutagenesis was also used to further dissect the structural elements that contribute to the interactions of the two MtNadD substrates, i.e. ATP and nicotinic acid mononucleotide (NaMN). For inhibitory profiling of partially active mutants and wild type MtNadD, we used a small molecule inhibitor of MtNadD with moderate affinity (Ki ∼ 25 μM) and antimycobacterial activity (MIC80) ∼ 40-80 μM). This analysis revealed interferences with some of the residues in the NaMN binding subsite consistent with the competitive inhibition observed for the NaMN substrate (but not ATP). A detailed steady-state kinetic analysis of MtNadD suggests that ATP must first bind to allow efficient NaMN binding and catalysis. This sequential mechanism is consistent with the requirement of transition to catalytically competent (open) conformation hypothesized from structural modeling. A possible physiological significance of this mechanism is to enable the down-regulation of NAD synthesis under ATP-limiting dormancy conditions. These findings point to a possible new strategy for designing inhibitors that lock the enzyme in the inactive over-closed conformation.


Lower extremity complex regional pain syndrome: long-term outcome after surgical treatment of peripheral pain generators.
We undertook a retrospective study to evaluate the hypothesis that complex regional pain syndrome (CRPS) I, known as the "new" reflex sympathetic dystrophy, persists because of undiagnosed injured joint afferents, cutaneous neuromas, or nerve compressions, and is, therefore, a misdiagnosed form of CRPS II, which is known as the "new" causalgia. We used a research protocol, with institutional review board approval, to review medical records for the purpose of identifying 30 patients with lower extremity reflex sympathetic dystrophy, based on their history, physical examination, neurosensory testing, and response to peripheral nerve blocks, who were treated surgically at the level of the peripheral nerve. In this report, we describe long-term outcomes in 13 of these patients who were followed up for a minimum of 24 months (mean, 47.8 months; range, 25-90 months). Based primarily on the results of physical examination and the response to peripheral nerve blocks, surgery included a combination of joint denervation, neuroma resection plus muscle implantation, and neurolysis. Outcomes were measured in terms of decreased pain medication usage and recovery of function, and the results were excellent in 7 (55%), good in 4 (30%), and poor (failure) in 2 (15%) of the patients. Based on these results, we concluded that most patients referred with a diagnosis of CRPS I have continuing pain input from injured joint or cutaneous afferents, and chronic nerve compression, which is indistinguishable from CRPS II, and amenable to successful treatment by means of an appropriate peripheral nerve surgical strategy.


Differential laminar distribution of corticostriatal neurons in the prefrontal and pericruciate gyri of the dog.
This investigation was undertaken to determine whether the laminar distribution of corticostriatal neurons differs between different anatomical and functional areas of the frontal lobe. Injections of lectin-conjugated horseradish peroxidase were made into the lateral, intermediate, and medial parts of the head of the caudate nucleus. After injections into the medial part of the caudate, most of the labeled neurons were found in the proreal and subproreal gyri of the prefrontal region. In both gyri, the majority of labeled cells were localized within layer V, and fewer neurons were located in the deeper part of layer III and in layers IV (proreal gyrus) and VI. No labeled neurons were noted in layer II. This distribution contrasted with that seen following injections into the intermediate and lateral parts of the caudate nucleus. In those cases, the majority of labeled cells were located in the pericruciate region (areas 4 and 6) and on the medial and lateral banks of the presylvian sulcus (paraorbital and internal composite areas, respectively). In the paraorbital and internal composite areas, the majority of labeled cells were still localized within layer V, but a larger percentage of labeled neurons were also noted in layer III. In areas 4 and 6, the laminar distribution shifted so that the majority of labeled cells were now located in layer III, with a widespread distribution of neurons throughout layers II-VI. These results indicate that corticostriatal neurons in the frontal lobe have different laminar distributions, depending on the particular anatomical or functional area in which they are located. The majority of neurons in the prefrontal region are located in infragranular layers, while those in the pericruciate region are located in supragranular layers.


Outer contour and radial changes of the cardiac left ventricle: a magnetic resonance imaging study.
Earlier studies have shown a +/-5% end-systolic decrease in the volume encompassed by the pericardial sack, manifesting as a radial diminution of the pericardial/epicardial contour of the left ventricle (LV). The aim of this study was to measure this radial displacement at different segmental levels of the LV and try to find out were it is as greatest and to calculate regional myocardial volume changes as a reference in healthy subjects. Eleven healthy subjects were examined by magnetic resonance imaging. Images were acquired using an ECG-triggered balanced fast field echo pulse sequence. The epicardial borders of the LV wall were delineated in end-diastole (ED) and end-systole (ES). Regional changes of the LV wall were analysed at three different levels (base, mid and apex) by dividing the myocardium into segments. The volumes obtained as the differences between the outer volume of the left ventricle at ED and ES at different slice levels were found to be greatest at the base of the heart and lowest at apex. The relative inward motion, that is the motion in short-axis direction of the epicardial border of the myocardium from ED to ES towards the centre of the LV, was greatest at the base and lowest at the mid level, something that has to be taken into account when measuring the LV during clinical exams. There was a significant difference in the relative inward motion between the segments at apex (p < 0.0001), mid (p = 0.036) and at base level (p < 0.0001).


The preprogrammed anti-inflammatory phenotypes of CD11chigh macrophages by Streptococcus pneumoniae aminopeptidase N safeguard from allergic asthma.
Early microbial exposure is associate with protective allergic asthma. We have previously demonstrated that Streptococcus pneumoniae aminopeptidase N (PepN), one of the pneumococcal components, inhibits ovalbumin (OVA) -induced airway inflammation in murine models of allergic asthma, but the underlying mechanism was incompletely determined. BALB/c mice were pretreated with the PepN protein and exposed intranasally to HDM allergen. The anti-inflammatory mechanisms were investigated using depletion and adoptive transfer experiments as well as transcriptome analysis and isolated lung CD11chigh macrophages. We found pretreatment of mice with PepN promoted the proliferation of lung-resident F4/80+CD11chigh macrophages in situ but also mobilized bone marrow monocytes to infiltrate lung tissue that were then transformed into CD11high macrophages. PepN pre-programmed the macrophages during maturation to an anti-inflammatory phenotype by shaping the metabolic preference for oxidative phosphorylation (OXPHOS) and also inhibited the inflammatory response of macrophages by activating AMP-activated protein kinase. Furthermore, PepN treated macrophages also exhibited high-level costimulatory signaling molecules which directed the differentiation into Treg. Our results demonstrated that the expansion of CD11chigh macrophages in lungs and the OXPHOS metabolic bias of macrophages are associated with reduced allergic airway inflammation after PepN exposure, which paves the way for its application in preventing allergic asthma.


Efficacy of tolfenamic acid and meloxicam in the control of postoperative pain following ovariohysterectomy in the cat.
The hypothesis was that Visual Analog Scale (VAS) scores would be lower, and mechanical wound thresholds (MWT) higher, in cats receiving tolfenamic acid compared to those receiving placebo in the postoperative period following elective ovariohysterectomy. Sixty-nine client-owned cats. A prospective, randomized, blinded and placebo-controlled study was performed in cats which underwent ovariohysterectomy following preoperative tolfenamic acid, meloxicam, or placebo. A second dose of the same analgesic was administered 24 hours postoperatively. Assessments were made 1-hour before induction and 1, 2, 4, 6, 22, and 25 hours postoperatively. Pain was assessed by a blinded observer using Numerical Rating (NRS) and VAS scales. The MWT were measured using a force-measuring device. Group comparison was performed by using one-way ANOVA and chi-squared test for qualitative and quantitative data, respectively, and a mixed model for repeated measurements (p < 0.05). Sixty-five cats were included in the study. There were no differences between groups at baseline. There was a treatment effect on the NRS scores at 6, 22 and 25 hours. The meloxicam group was less painful than controls at 6 and 22 hours; both treatment groups were less painful than controls at 25 hours. There were no differences between groups in VAS for pain or sedation. The number of animals receiving rescue analgesia did not differ between groups. There was a treatment effect on MWT; thresholds in both treatment groups were significantly higher than that observed in controls at all time points. Preoperative tolfenamic acid or meloxicam reduced wound sensitivity following ovariohysterectomy in the cat. Tolfenamic acid and meloxicam administered preoperatively provided a similar analgesic effect in the postoperative period lasting 24 hours. Mechanical thresholds may be a better way of evaluating postoperative analgesia provided by nonsteroidal anti-inflammatory drugs in cats.


The role of religious values in coping with cancer.
The relationship between transcendent meaning attribution, religious orientation, and psychological well-being was studied in cancer and noncancer patients to test the hypotheses that intrinsic religious values and life meaning enhance coping and well-being during the course of the life-threatening illness. Subjects were 44 patients receiving medical treatment for cancer and noncancer medical conditions. In the cancer group, higher levels of attributed life meaning were positively linked with intrinsic religious orientation, and associated with lower levels of despiar, anger-hostility, and social isolation. Cancer patients scored higher than noncancer patients on depersonalization, suggesting the presence of psychic numbing in response to their illness. Noncancer group results were characterized by positive correlations between the two groups in coping styles and salience of life meaning attribution. A rationale for the observed differences in coping styles between the two groups is presented, highlighting perceived life threat as a key differentiating variable.


Colonization, biofilm formation and biodegradation of polyethylene by a strain of Rhodococcus ruber.
A two-step enrichment procedure led to the isolation of a strain of Rhodococcus ruber (C208) that utilized polyethylene films as sole carbon source. In liquid culture, C208 formed a biofilm on the polyethylene surface and degraded up to 8% (gravimetrically) of the polyolefin within 30 days of incubation. The bacterial adhesion to hydrocarbon assay and the salt aggregation test both showed that the cell-surface hydrophobicity of C208 was higher than that of three other isolates which were obtained from the same consortium but were less efficient than C208 in the degradation of polyethylene. Mineral oil, but not nonionic surfactants, enhanced the colonization of polyethylene and increased biodegradation by about 50%. Fluorescein diacetate (FDA) hydrolysis and protein content analysis were used to test the viability and biomass density of the C208 biofilm on the polyethylene, respectively. Both FDA activity and protein content of the biofilm in a medium containing mineral oil peaked 48-72 h after inoculation and then decreased sharply. This finding apparently reflected rapid utilization of the mineral oil adhering to the polyethylene. The remaining biofilm population continued to proliferate moderately and presumably played a major role in biodegradation of the polyethylene. Fourier transform infrared spectra of UV-photooxidized polyethylene incubated with C208 indicated that biodegradation was initiated by utilization of the carbonyl residues formed in the photooxidized polyethylene.


A novel protein RASON encoded by a lncRNA controls oncogenic RAS signaling in KRAS mutant cancers.
Mutations of the RAS oncogene are found in around 30% of all human cancers yet direct targeting of RAS is still considered clinically impractical except for the KRASG12C mutant. Here we report that RAS-ON (RASON), a novel protein encoded by the long intergenic non-protein coding RNA 00673 (LINC00673), is a positive regulator of oncogenic RAS signaling. RASON is aberrantly overexpressed in pancreatic ductal adenocarcinoma (PDAC) patients, and it promotes proliferation of human PDAC cell lines in vitro and tumor growth in vivo. CRISPR/Cas9-mediated knockout of Rason in mouse embryonic fibroblasts inhibits KRAS-mediated tumor transformation. Genetic deletion of Rason abolishes oncogenic KRAS-driven pancreatic and lung cancer tumorigenesis in LSL-KrasG12D; Trp53R172H/+ mice. Mechanistically, RASON directly binds to KRASG12D/V and inhibits both intrinsic and GTPase activating protein (GAP)-mediated GTP hydrolysis, thus sustaining KRASG12D/V in the GTP-bound hyperactive state. Therapeutically, deprivation of RASON sensitizes KRAS mutant pancreatic cancer cells and patient-derived organoids to EGFR inhibitors. Our findings identify RASON as a critical regulator of oncogenic KRAS signaling and a promising therapeutic target for KRAS mutant cancers.


Matrix metalloproteinase and alphavbeta3 integrin-dependent vascular smooth muscle cell invasion through a type I collagen lattice.
Smooth muscle cell (SMC) migration from the tunica media to the intima is a key event in the development of atherosclerotic lesions and in restenosis after angioplasty. SMCs require not only migratory but also degradative abilities that enable them to migrate through extracellular matrix proteins, which surround and embed these cells. We used a collagen type I lattice as a coating on top of a porous filter as a matrix barrier in a chamber to test the invasive behavior of SMCs in response to a chemoattractant (invasion assay) and compared that behavior with simple SMC migration through collagen type I-coated filters (migration assay). Inhibitors of matrix metalloproteinase, KB-R8301, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), TIMP-2, and peptide 74, attenuated platelet-derived growth factor-BB (PDGF-BB)-directed SMC invasion across the collagen lattice, whereas no effect was seen with these inhibitors on simple SMC migration through collagen-coated filters. RGD peptide inhibited SMC invasion but did not affect SMC migration. Anti-alphavbeta3 integrin antibody attenuated PDGF-BB-directed SMC invasion, whereas other antibodies against RGD-recognizing integrins, namely alphavbeta5 and alpha5, had no effect. None of these antibodies had any effect on simple SMC migration. RGD peptide and anti-alphavbeta3 antibody inhibited the attachment and spreading of SMCs on denatured collagen but not on native collagen. These findings indicate that there is a difference in the mechanisms between simple SMC migration across a collagen-coated filter and SMC invasion through a fibrillar collagen barrier. A proteolytic process is required for SMC invasion, and the degradation of matrix proteins alters the relationship between matrix protein molecules and SMC surface integrins.


Two courses of four weekly infusions of rituximab with or without interferon-α2a: final results from a randomized phase III study in symptomatic indolent B-cell lymphomas.
Patients with advanced CD20 + indolent lymphoma, requiring therapy, were randomized to rituximab (four weekly infusions of 375 mg/m(2)) or to rituximab combined with 5 weeks of interferon-α2a (IFN-α2a) (3-4.5 MIU daily) as priming. Responding patients were eligible for a second cycle with the same allocated treatment. In total, 156 patients were randomized to rituximab and 157 to rituximab + IFN-α2a. In the intention-to treat (ITT) population, 244 patients (78%) responded to cycle 1. After a second cycle the complete remission/complete remission unconfirmed (CR/CRu) rate was 41% with the combination versus 24% with monotherapy (p = 0.005). The median time to treatment failure (primary endpoint) in ITT patients was 28 vs. 21.5 months, respectively (p = 0.302). After a long median follow-up (61 months), 33% (42% of patients responding to cycle 1) were still failure-free with an overall survival rate of 88% and with no difference between the treatment groups. The trial was registered at ClinicalTrials.gov Identifier: NCT01609010.


Member and dentist narratives about a dental programme for the Medicaid expansion population: A content analysis.
In the United States, the 2010 Affordable Care Act's Medicaid expansion improved financial access to dental care for new Medicaid enrollees in states with Medicaid dental benefits for adults. Using an existing theoretical framework, we explored dentist and Medicaid member narratives concerning a state's redesigned dental programme for the Medicaid expansion population. Data sources include two surveys conducted in 2016-one to a random sample of Medicaid members and one to private practice dentists in Iowa. A qualitative content analysis approach was used to quantify and describe themes present in open-ended comment data. The most common themes among Medicaid member narratives were related to oral health needs and status, availability of dentists who accept Medicaid, and Medicaid coverage for dental care. Among dentists, the programme's administrative burden was the most common theme, with coverage and reimbursement commonly cited as well. An emergent theme among both member and dentist comments was patient confusion or lack of knowledge about programme enrolment and coverage. Member awareness about enrolment and coverage in public dental benefit plans should be further explored to determine the impact on access to care. Effectively communicating the intent, programme design and expectations of a Medicaid programme to both providers and Medicaid members is important to the success of the programme.


Fontan conversion: identifying the high-risk patient.
Patients with atriopulmonary Fontan tend to undergo conversion to total cavopulmonary connections secondary to arrhythmias or poor flow dynamics. However, the ideal candidate is unknown. Between December 1994 and May 2011, 70 patients (40 males [57%]) underwent Fontan conversion. Median age was 23 years (range, 4 to 46 years). Excluded were 1.5 ventricle conversions. The most common diagnoses included tricuspid atresia in 34 patients (49%) and double-inlet left ventricle in 16 (23%). Atrial tachyarrhythmia was present in 62 patients (89%), 41 (59%) had atrioventricular valve (AVV) regurgitation, and 32 (46%) were in New York Heart Association class III or IV. Atriopulmonary Fontan was the original connection in 58 patients (83%), whereas the Björk modification was performed in 8 (11%). Fontan was performed with an intraatrial conduit in 41 patients, an extracardiac conduit in 18, and a lateral tunnel in 11. Forty-nine patients (70%) underwent concomitant arrhythmia operations. Early death occurred in 10 patients (14%). Multivariate analysis revealed age older than 27 years (p = 0.009), AVV regurgitation (p = 0.016), lack of arrhythmia operation (p = 0.04), and male sex (p = 0.02) were predictors of perioperative death. Mean follow-up was 5 years (maximum, 17 years). Overall survival at 1, 5, and 10 years was 81%, 70%, and 67%, respectively, and 84% of patients were in New York Heart Association class I or II. Proper selection of Fontan conversion candidates is critical. Concomitant arrhythmia operations may be associated with improved survival. Older age and AVV regurgitation increase the risk of poor outcome, and cardiac transplantation may be a better option.


Urban and rural patterns of bicycle helmet use: factors predicting usage.
To document current bicycle helmet use in Winnipeg, Manitoba and nearby rural communities, and to identify target groups for a helmet promotion campaign. Cyclist helmet use was observed between 28 May and 20 August 1996 at a sample of urban and rural locations. Age, gender, helmet use, riding companion(s), location type, correct helmet use, and use of headphones were recorded. Univariate and multivariate analyses were performed. Adjusted odds ratios with 95% confidence intervals were calculated from the final models. Altogether 2629 cyclists (70% male, 30% female) were observed: 2316 at 183 urban locations and 313 at 25 rural locations, with nearly equal numbers of children and adults observed. Overall helmet use was 21.3%, with lower use in males (18.9%) than females (26.3%), despite gender only being a significant variable on multivariate analysis for children under 8 years and adults. Urban helmet use was considerably higher (22.9%) than rural use (8.9%). Helmet use increased linearly as mean neighbourhood income increased, with a nearly fourfold difference in use between the highest and lowest income neighbourhoods. Children less than 8 years old and adults had the highest, and teenagers the lowest, use. Significant predictive variables were identified separately by age category to inform targeted programming. We documented low helmet use in our region, emphasizing the need for a regional helmet promotion campaign as well as future helmet legislation. A marked urban-rural difference in helmet use that has not been previously reported was also identified. Target groups for a future campaign include adolescents, males, rural cyclists, and those in lower income neighbourhoods.


Osmolyte homeostasis controls single-cell growth rate and maximum cell size of Saccharomyces cerevisiae.
Cell growth is well described at the population level, but precisely how nutrient and water uptake and cell wall expansion drive the growth of single cells is poorly understood. Supported by measurements of single-cell growth trajectories and cell wall elasticity, we present a single-cell growth model for yeast. The model links the thermodynamic quantities, such as turgor pressure, osmolarity, cell wall elasto-plasticity, and cell size, applying concepts from rheology and thin shell theory. It reproduces cell size dynamics during single-cell growth, budding, and hyper-osmotic or hypo-osmotic stress. We find that single-cell growth rate and final size are primarily governed by osmolyte uptake and consumption, while bud expansion requires additionally different cell wall extensibilities between mother and bud. Based on first principles the model provides a more accurate description of size dynamics than previous attempts and its analytical simplification allows for easy combination with models for other cell processes.


Combining quantitative and qualitative measures of uncertainty in model-based environmental assessment: the NUSAP system.
This article discusses recent experiences with the Numeral Unit Spread Assessment Pedigree (NUSAP) system for multidimensional uncertainty assessment, based on four case studies that vary in complexity. We show that the NUSAP method is applicable not only to relatively simple calculation schemes but also to complex models in a meaningful way and that NUSAP is useful to assess not only parameter uncertainty but also (model) assumptions. A diagnostic diagram can be used to synthesize results of quantitative analysis of parameter sensitivity and qualitative review (pedigree analysis) of parameter strength. It provides an analytic tool to prioritize uncertainties according to quantitative and qualitative insights in the limitations of available knowledge. We show that extension of the pedigree scheme to include societal dimensions of uncertainty, such as problem framing and value-laden assumptions, further promotes reflexivity and collective learning. When used in a deliberative setting, NUSAP pedigree assessment has the potential to foster a deeper social debate and a negotiated management of complex environmental problems.


Detection of HTLV-1 by polymerase chain reaction in situ hybridization in adult T-cell leukemia/lymphoma.
A method for nonradioactive polymerase chain reaction in situ hybridization was developed and used to determine the distribution of human T-lymphotropic virus type I (HTLV-I) proviral DNA in paraffin-embedded surgical specimens of adult T-cell leukemia/lymphoma (ATLL). As controls, we used biopsy samples of five cases of mycosis fungoides, cells of an HTLV-I-infected cell line (MT2), as well as HTLV-1-negative cells (YAS). We successfully detected the amplicon of the HTLV-1 tax sequence in the nuclei of the cutaneous infiltrating lymphoid cells in 90% (9/10) of ATLL cases. Studies also revealed the existence of HTLV-1 provirus DNA in nuclei of sweat gland epithelial cells and vascular endothelial cells as well as lymphoid cells in ATLL patients. Mycosis fungoides and YAS cells were negative for the HTLV-I tax sequence, but MT2 cells were strongly positive. The results indicated that this technique was more sensitive in detecting intracellular amplicons than was the previous in situ hybridization method. Through its use, we were able to easily determine the distribution of HTLV-I-positive cells among the various cells and tissues of paraffin-embedded archival materials.


Preparation of a novel streptokinase mutant with improved stability.
The novel mutant streptokinase, SK-K59E, can activate human plasminogen as efficiently as the purified commercially available streptokinase. Several peptide bonds including Lys59-Ser60 in native streptokinase were hydrolyzed in reaction with plasmin and peptides of small molecular masses were generated. The plasminogen activator activity of native streptokinase in reaction with human plasmin declined to 25% of the original activity in a 120-min incubation. On the other hand, the NH2-terminal peptide of SK-K59E remained intact in reaction with plasmin and the activator activity of streptokinase decreased to 75% of the original activity in 120 min. The major degraded peptide fragments of native streptokinase in reaction with plasmin had molecular masses of 36 and 30 kDa. However, two major peptide fragments of 42 and 34 kDa were observed in the reaction of SK-K59E with human plasmin. The 42 kDa peptide fragment, which contained NH2-terminal of streptokinase, could activate human plasminogen as efficiently as the native streptokinase. SK-K59E can induce greater degree of caseinolysis and fibrinolysis than the native streptokinase. In conclusion, the results demonstrate that the prevention of cleavage at Lys59 of streptokinase prolongs the half-life of streptokinase in complex with plasmin and that the NH2-terminal of streptokinase (Ile1-Lys59) plays an important role in maintaining its stability.


Pathologic continuum of acute appendicitis: sonographic findings and clinical management implications.
Appendicitis is one of the most common causes of the acute abdomen often requiring emergent surgery. Delayed diagnosis leads to the progression of uncomplicated appendicitis to complicated (gangrenous, perforated) appendicitis, often changing clinical management. Computed tomography and ultrasound are imaging modalities of choice to preoperatively diagnose appendicitis. Recent concerns of radiation exposure and cost have renewed interest in using ultrasound as an initial, diagnostic study. A sonographic pictorial and histopathologic review of the continuum of appendicitis is presented. A comprehensive sonographic examination of the appendix should investigate the size (maximal diameter), the echogenic submucosal layer integrity, the mural color Doppler signature, the presence of a fecalith, and the periappendiceal changes. Features of an uncomplicated appendicitis include size greater than 6 to 7 mm, hyperemia on color Doppler, mural thickening, and an intact echogenic submucosal layer. Gangrenous appendicitis is characterized by loss of the echogenic submucosal layer with absent color Doppler flow. Loculated pericecal fluid, prominent pericecal fat, and circumferential loss of the submucosal layer are suggestive of perforation. Sonographic staging can triage management of appendicitis by directing urgent laparoscopic appendectomy for uncomplicated appendicitis, open appendectomy for complicated appendicitis, and conservative management (antibiotics with percutaneous drainage) for perforated appendicitis with abscess formation.


Long-term subcutaneous morphine administration after surgery in newborns.
To analyze the management of newborns after major surgery receiving morphine subcutaneously and to identify possible side effects. Morphine was administered via a subcutaneous catheter (Insuflon) in 20 newborns after major surgery. Side effects like hypotension, pain during morphine administration and local infection were noted. Morphine dose was adjusted according to the hospital guidelines with the Neonatal Infant Pain Score (NIPS) and the Finnegan withdrawal score. Surgery was performed at the median age of 38 5/7 weeks (range: 32 1/7-49 5/7 weeks). Before starting subcutaneous morphine administration, patients received intravenous morphine for a median of two weeks (range six days to seven weeks). All patients showed good pain relief with no severe side effects. Three patients reacted with crying to the first dose of subcutaneous morphine. No other side effects occurred. Subcutaneous application of morphine with the Insuflon catheter is an alternative to intravenous treatment of postoperative pain in neonates. In this small group pain relief was good and side effects were harmless.


The effects of plumbous ion on protein biosynthesis in reticulocytes.
Plumbous ion a potent inhibitor of hemoglobin synthesis by reticulocytes in vivo, is not as effective in inhibiting in vitro globin synthesizing systems prepared from these cells. The reticulocyte is considerably more sensitive to Pb2+ than are leukemic leukocytes, HeLa cells or bacteria. In fact, protein synthesis in leukemic leukocytes is actually stimulated by Pb2+. The synthesis of non-heme protein as well as hemoglobin is inhibited by Pb2+ in reticulocytes and the synthesis of alpha chains is inhibited to a greater degree tcids into reticulocytes. All of these observations are consistent with the biosynthesis of heme rather than any of the steps in protein biosynthesis being the locus for inhibition of hemoglobin synthesis by low levels of plumbous ion. Also, there is marked biological variations in the susceptibilities of reticulocytes from different rabbits to Pb2+.


The impact of PNPLA3 (rs738409 C>G) polymorphisms on liver histology and long-term clinical outcome in chronic hepatitis B patients.
We aimed to assess the association between the patatin-like phospholipase domain-containing-3 (PNPLA3) I148M polymorphism, liver histology and long-term outcome in chronic hepatitis B (CHB) patients. We enrolled 531 consecutive treatment naïve CHB patients diagnosed from 1985 to 2012 with an available liver biopsy for reassessment, and sample for genetic testing. Data on all-cause mortality and hepatocellular carcinoma (HCC) at long-term follow-up were obtained from national database registries. The prevalence of steatohepatitis increased with PNPLA3 CC (14%), CG (20%) and GG (43%) (P < 0.001). The association was altered by both gender (P = 0.010) and overweight (P = 0.015): the effect of PNPLA3 on steatohepatitis was most pronounced among non-overweight females (adjusted OR 13.4, 95%CI: 3.7-51.6, P < 0.001), and non-overweight males (adjusted OR 2.4, 95%CI: 1.4-4.3, P = 0.002). Furthermore, PNPLA3 GG genotype was associated with iron depositions (OR 2.8, 95%CI: 1.2-6.4, P = 0.014) and lobular inflammation (OR 2.2, 95%CI: 1.1-4.5, P = 0.032), but not with advanced fibrosis (OR 1.1, 95%CI: 0.7-1.8, P = 0.566). The median follow-up was 10.1 years (interquartile range 5.6 - 15.8), during which 13 patients developed HCC and 28 died. Steatohepatitis was associated with all-cause mortality [Hazard ratio (HR) 3.1, 95%CI: 1.3-7.3, P = 0.006] and HCC (HR 2.8, 95%CI: 0.9-9.2, P = 0.078), but no significant association was observed for PNPLA3. In this cohort of biopsied CHB patients, PNPLA3 was independently associated with steatosis, steatohepatitis, lobular inflammation and iron depositions, but not with advanced fibrosis, HCC development or all-cause mortality. The effect of PNPLA3 on steatohepatitis was particularly pronounced among female patients without severe overweight.


Self-reported Conflicts of Interest and Trial Sponsorship of Clinical Trials in Prostate Cancer Involving Radiotherapy.
To examine the association between trial sponsorship and conflicts of interest (COI) with clinical trial conclusions for prostate cancer trials related to radiotherapy. The MEDLINE database was searched for all prostate cancer clinical trials published between 2004 and 2013 and identified 1396 studies. Two investigators independently identified trials published in the English language of ≥30 patients, and extracted relevant data. Clinical trials were classified according to trial characteristics, sponsorship source and type, COI, and study conclusion, and analyzed by univariable and multivariable logistic regression. Of 240 eligible trials, 160 (67.5%) evaluated drugs without radiotherapy, 60 (25%) involved radiotherapy, and 18 (7.5%) involved procedures without radiotherapy. Of the 60 radiotherapy trials eligible for analysis, positive sponsorship and potential COI were present in 58.3% and 20% of trials, respectively. Study conclusions were positive, negative, or neutral in 78.3%, 5%, and 16.7% of trials, respectively. No association was found between positive conclusions and either industry support of potential COI. Positive conclusions were reported in 86.7% and 83.3% of trials with sponsorship and COI, respectively, as compared with 75.6% and 77.1% of those without sponsorship (P=0.37) and COI (P=0.64). Sponsorship was significantly associated with radiotherapy trials combined with drugs (odds ratio 5.5, P=0.01) and higher-risk disease (odds ratio 4.71, P=0.01). The presence of sponsorship was associated with radiotherapy trials involving drugs or studying higher-risk prostate cancer. However, there were no identified associations between study conclusion and sponsorship type or COI.


Effect of citric acid supplements to a maize-soya diet on the in vitro availability of minerals, trace elements, and heavy metals.
The aim of the present study was to evaluate the effect of increasing supplements of citric acid (CTA) on the in vitro availability of mineral (Ca, Mg), trace elements (Mn, Zn) and heavy metals (Pb, Cd). Phytaterich diets (0.8%) based on maize, soybean meal, corn starch and soybean oil (containing 5.0 g Ca, 1.2 g Mg, 50 mg Mn, 22 mg Zn, 10 mg Pb and 5 mg Cd per kg diet) were supplemented with 0, 1, 2, 3 and 4% of CTA. All diets were treated in a multi-enzyme in vitro system followed by equilibrium dialysis after peptic and pancreatic digestion. Under the conditions investigated, dietary addition of CTA enhanced dialysability of calcium, magnesium, manganese and zinc. Furthermore, a dose-dependent increase of lead and cadmium dialysability due to CTA supplementations was evident.


Primary Metabolites, Anthocyanins, and Hydrolyzable Tannins in the Pomegranate Fruit.
Pomegranate (Punica granatum L.) is an important and interesting fruit tree that is cultivated in many parts of the world. In recent years, along with the increase in its cultivation and consumption there has been a dramatic increase in the scientific interest in its biology, methods of cultivation, adaptation to environmental cues and its health-promoting properties. Quite a large proportion of the various metabolites produced in the pomegranate were determined and their content in the bark, roots, leaves, and fruit was reported. Many reviews on polyphenolic compound content, antioxidant activity and health-promoting compounds were published recently. However, only very few recent reports were dedicated to primary metabolites, despite the fact that much work was done on organic acids, sugars, proteins, lipids, and amino acids of the pomegranate fruit. In this review, a special effort was made to present these recent studies and the review is devoted to primary metabolites. The reported data show high variation in the content of primary metabolites within the pomegranate fruit; therefore the data is presented (whenever possible) according to fruit tissues (peel, arils, and seeds), developmental stages of the fruit, environmental and climatic conditions, and genetic background. Most of the data on pomegranate is based on metabolic content and contains no genetic or molecular analysis except for work done on anthocyanins and hydrolyzable tannins. In those cases, gene assignment and genetic control studies were pointed out in the review. The recent publication of the genome sequences from several pomegranate varieties and transcriptomic data from fruits, flowers, and leaves is expected to facilitate the understanding of genetic control of metabolites in pomegranate.


Prognostic value of neutrophil count to albumin ratio in patients with decompensated cirrhosis.
Our study aimed to investigate the prognostic value of neutrophil count to albumin ratio (NAR) in predicting short-term mortality of patients with decompensated cirrhosis (DC). A total of 623 DC patients were recruited from a retrospective observational cohort study. They were admitted to our hospital from January 2014 to December 2015. NAR of each patient was calculated and analyzed for the association with 90-day liver transplantation-free (LT-free) outcome. The performance of NAR and the integrated model were tested by a receiver-operator curve (ROC) and C-index. The 90-day LT-free mortality of patients with DC was 10.6%. NAR was significantly higher in 90-day non-survivors than in survivors (The median: 1.73 vs 0.76, P < 0.001). A threshold of 1.40 of NAR differentiated patients with a high risk of death (27.45%) from those with a low risk (5.11%). By multivariate analysis, high NAR was independently associated with poor short-term prognosis (high group: 5.07 (2.78, 9.22)). NAR alone had an area under the ROC curve of 0.794 and C-index of 0.7789 (0.7287, 0.8291) in predicting 90-day mortality. The integrated MELD-NAR (iMELD) model had a higher area under the ROC (0.872) and C-index (0.8558 (0.8122, 0.8994)) than the original MELD in predicting 90-day mortality. NAR can be used as an independent predictor of poor outcomes for patients with DC during short-term follow-up.


Needle-free jet injection for administration of influenza vaccine: a randomised non-inferiority trial.
Administration of vaccines by needle-free technology such as jet injection might offer an alternative to needles and syringes that avoids the issue of needle phobia and the risk of needle-stick injury. We aimed to assess the immunogenicity and safety of trivalent influenza vaccine given by needle-free jet injector compared with needle and syringe. For this randomised, comparator-controlled trial, we randomly assigned (1:1) healthy adults (aged 18-64 years) who attended one of four employee health clinics in the University of Colorado health system, with stratification by site, to receive one dose of the trivalent inactivated influenza vaccine Afluria given either intramuscularly with a needle-free jet injector (Stratis; PharmaJet, Golden, CO, USA) or with needle and syringe. Randomisation was done with a computer-generated randomisation schedule with a block size of 100. Because of the nature of the study, masking of participants was not possible. Immunogenicity was assessed by measurement of the hemagglutination inhibition antibody titres in serum for the three viral strains included in the vaccine. We included six coprimary endpoints: three strain-specific geometric mean titre ratios and the absolute differences in three strain-specific seroconversion rates. The immune response of the jet injector group was regarded as non-inferior to that of the needle and syringe group if both the upper bound of each of the three 95% CIs for the strain-specific geometric mean titre ratios was 1.5 or less, and the upper bound of the three 95% CIs for the strain-specific seroconversion rate differences was less than 10 percentage points. We used t test for group comparison. This study is registered with ClinicalTrials.gov, number NCT01688921. During the 2012-13 influenza season of the northern hemisphere, we allocated 1250 participants to receive vaccination by needle-free jet injector (n=627) or needle and syringe (n=623). In the intention-to-treat immunogenicity population, all participants with two serum samples were included (575 in the jet injector group and 574 in the needle and syringe group). The immune response to Afluria when given by needle-free jet injector met the criteria for non-inferiority for all six coprimary endpoints. The jet injector group met the geometric mean titre criterion for non-inferiority for the A/H1N1, A/H3N2, and B strains (upper bound of the 95% CI for the geometric mean titre ratios were 1·10 for A/H1N1, 1·17 for A/H3N2, and 1·04 for B strains). The jet injector group met the seroconversion rate criterion for non-inferiority for the A/H1N1, A/H3N2, and B strains (upper bound of the 95% CI of the seroconversion rate differences were 6·0% for A/H1N1, 7·0% for A/H3N2, and 5·7% for B strains). We recorded serious adverse events in three participants, none of which were study related. The immune response to influenza vaccine given with the jet injector device was non-inferior to the immune response to influenza vaccine given with needle and syringe. The device had a clinically acceptable safety profile, but was associated with a higher frequency of local injection site reactions than was the use of needle and syringe. The Stratis needle-free jet injector device could be used as an alternative method of administration of Afluria trivalent influenza vaccine. Biomedical Advanced Research and Development Authority (BARDA), PATH, bioCSL, and PharmaJet.


Safety and reliability of femtosecond laser-assisted cataract surgery for Japanese eyes.
To assess the safety and efficacy of femtosecond laser-assisted cataract surgery (FLACS) in Japanese eyes. Retrospective case series. The clinical records of 529 consecutive eyes of 312 Japanese patients who underwent FLACS from 2013 to 2016 were reviewed. The completion rates of anterior capsulotomy and corneal incisions using the femtosecond laser (LenSx, Alcon) and the number of attempts to achieve secure docking between the cornea and laser system were recorded. The uncorrected and corrected distance visual acuities (VAs) and uncorrected and distance-corrected near VAs 1 week postoperatively were evaluated. Anterior capsulotomies were completed in 98.7% of eyes. The corneal incisions were incomplete in 8.3%, which included eyes with arcus senilis and neovascularization. The mean corneal endothelial cell reduction rate was 5.5%. Multiple docking attempts were required in 22.9% of eyes, while canthotomy was performed in two eyes. The postoperative VAs were favorable. FLACS was safe and reliable in Japanese eyes. Further improvements in docking are required to accommodate small corneas and narrow palpebral spaces.


Pravastatin limits endothelial activation after irradiation and decreases the resulting inflammatory and thrombotic responses.
Endothelial dysfunction has been implicated in the pathogenesis of atherosclerosis, fibrosis and vascular occlusion after radiation therapy. Statins have been reported to improve endothelial function; however, this beneficial effect on endothelial cells has never been investigated after irradiation. Therefore, using human microvascular endothelial cells from lung that had been irradiated with 5 or 10 Gy, we assessed the effect of pravastatin on endothelial activation by ELISA, cell-ELISA and electrophoretic mobility shift assay and increased blood-endothelial cell interactions by a flow adhesion assay. Pravastatin inhibited the overproduction of monocyte chemoattractant protein 1, IL6 and IL8 and the enhanced expression of intercellular adhesion molecule 1 but had no effect on platelet-endothelial cell adhesion molecule 1 expression. Moreover, pravastatin down-regulated the radiation-induced activation of the transcription factor activator protein 1 but not of nuclear factor-kappaB. Finally, an inhibition by pravastatin of increased adhesion of leukocytes and platelets to irradiated endothelial cells was observed. The effect of pravastatin was maintained up to 14 days after irradiation and was reversed by mevalonate. Pravastatin exerts persistent anti-inflammatory and anti-thrombotic effects on irradiated endothelial cells. Statins may be considered in therapeutic strategies for the management of patients treated with radiation therapy.


Protein Kinase C-θ (PKC-θ) in Natural Killer Cell Function and Anti-Tumor Immunity.
The protein kinase C-θ (PKCθ), which is essential for T cell function and survival, is also required for efficient anti-tumor immune surveillance. Natural killer (NK) cells, which express PKCθ, play a prominent role in this process, mainly by elimination of tumor cells with reduced or absent major histocompatibility complex class-I (MHC-I) expression. This justifies the increased interest of the use of activated NK cells in anti-tumor immunotherapy in the clinic. The in vivo development of MHC-I-deficient tumors is much favored in PKCθ(-/-) mice compared with wild-type mice. Recent data offer some clues on the mechanism that could explain the important role of PKCθ in NK cell-mediated anti-tumor immune surveillance: some studies show that PKCθ is implicated in signal transduction and anti-tumoral activity of NK cells elicited by interleukin (IL)-12 or IL-15, while others show that it is implicated in NK cell functional activation mediated by certain killer-activating receptors. Alternatively, the possibility that PKCθ is involved in NK cell degranulation is discussed, since recent data indicate that it is implicated in microtubule-organizing center polarization to the immune synapse in CD4(+) T cells. The implication of PKC isoforms in degranulation has been more extensively studied in cytotoxic T lymphocyte, and these studies will be also summarized.


In search of lost time: Reconstructing the unfolding of events from memory.
When remembering an event, not only do we recollect what happened, when and where it happened, but also how it unfolded over time. What aspects of events are encoded in memory to support this recollection? This question is central for understanding the nature of event memories and our reconstruction of the time passed. In this article, we investigate how the spontaneous encoding of unfamiliar animations during learning influences the recollection of how these animations unfold. Specifically, we examine two structural properties of dynamic event sequences known to modulate the amount of information encoded in memory: the perceived number of sub-events and their perceived similarity. We found that despite clock duration remaining constant, more sub-events and less similar ones led to longer recognition memory latencies, duration judgments and mental event replaying. In particular, across stimulus animations, both the perceived number of sub-events and their degree of similarity contributed to the prediction of duration judgments and the length of mental event reproductions. Results indicate that the number and nature of sub-events in a sequence modulate how we reconstruct its duration and temporal unfolding, thus suggesting that these event properties, which mediate the amount of information encoded for an event, modulate the subsequent recollection of its temporal unfolding.


The Impact of Job-Demand-Control-Support on Leptin and Ghrelin as Biomarkers of Stress in Emergency Healthcare Workers.
Despite the available literature on the consequences of night shiftwork on stress and food intake, its impact on leptin and ghrelin has never been studied. We previously demonstrated that leptin and ghrelin were biomarkers related to stress, and acute stress-induced a decrease in leptin levels and an increase in ghrelin levels. We performed a prospective observational study to assess the influence of night work, nutrition, and stress on the levels of ghrelin and leptin among emergency healthcare workers (HCWs). We took salivary samples at the beginning of a day shift and/or at the end of a night shift. We also monitored stress using the job demand-control-support model of Karasek. We recorded 24-h food intake during the day shift and the consecutive night shift and during night work and the day before. We included 161 emergency HCWs. Emergency HCWs had a tendency for decreased levels of leptin following the night shift compared to before the dayshift (p = 0.067). Furthermore, the main factors explaining the decrease in leptin levels were an increase in job-demand (coefficient -54.1, 95 CI -99.0 to -0.92) and a decrease in job control (-24.9, -49.5 to -0.29). Despite no significant changes in ghrelin levels between shifts, social support was the main factor explaining the increase in ghrelin (6.12, 0.74 to 11.5). Food intake (kcal) also had a negative impact on leptin levels, in addition to age. Ghrelin levels also decreased with body mass index, while age had the opposite effect. In conclusion, we confirmed that ghrelin and leptin as biomarkers of stress were directly linked to the job demand-control-support model of Karasek, when the main cofounders were considered.


Isolation of PCV3 from Perinatal and Reproductive Cases of PCV3-Associated Disease and In Vivo Characterization of PCV3 Replication in CD/CD Growing Pigs.
Porcine circovirus 3 (PCV3) has been identified as a putative swine pathogen with a subset of infections resulting in stillborn and mummified fetuses, encephalitis and myocarditis in perinatal, and periarteritis in growing pigs. Three PCV3 isolates were isolated from weak-born piglets or elevated stillborn and mummified fetuses. Full-length genome sequences from different passages and isolates (PCV3a1 ISU27734, PCV3a2 ISU58312, PCV3c ISU44806) were determined using metagenomics sequencing. Virus production in cell culture was confirmed by qPCR, IFA, and in situ hybridization. In vivo replication of PCV3 was also demonstrated in CD/CD pigs (n = 8) under experimental conditions. Viremia, first detected at 7 dpi, was detected in all pigs by 28 dpi. IgM antibody response was detected between 7-14 dpi in 5/8 PCV3-inoculated pigs but no IgG seroconversion was detected throughout the study. Pigs presented histological lesion consistent with multi systemic inflammation characterized by myocarditis and systemic perivasculitis. Viral replication was confirmed in all tissues by in situ hybridization. Clinically, all animals were unremarkable throughout the study. Although the clinical relevance of PCV3 remains under debate, this is the first isolation of PCV3 from perinatal and reproductive cases of PCV3-associated disease and in vivo characterization of PCV3 infection in a CD/CD pig model.


[A new method of sterilization of donor eyes].
Microbiologic examinations of 200 pairs of donor eyes have revealed Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, and Proteus infection of 124 pairs of eyes (62 percent). Sensitivities of the isolated microorganisms to various antibacterial drugs was examined. Gentamicin and chlorhexidine proved to be the most effective in respect of the microorganisms isolated from cadaveric eyes. These results enabled the authors suggest a new method for donor eyes sterilization, consisting in placing the donor eyes in aqueous solution of chlorhexidine (0.05 percent) and gentamicin (0.0125 percent) for 10 min immediately before surgery. This method was tried with 300 pairs of cadaveric eyes. Its high efficacy was confirmed by the absence of postoperative complications induced by infection transmission via the transplant. Early infectious complications were recorded in 8 patients of the reference group, where the transplants were sterilized routinely; in 4 cases it was a P. aeruginosa infection.


Small bioactive molecules as dual functional co-dopants for conducting polymers.
Biological responses to neural interfacing electrodes can be modulated via biofunctionalisation of conducting polymer (CP) coatings. This study investigated the use of small bioactive molecules with anti-inflammatory properties. Specifically, anionic dexamethasone phosphate (DP) and valproic acid (VA) were used to dope the CP poly(ethylenedioxythiophene) (PEDOT). The impact of DP and VA on material properties was explored both individually and together as a codoped system, compared to the conventional dopant p-toluenesulfonate (pTS). Electrical properties of DP and VA doped PEDOT were reduced in comparison to PEDOT/pTS, however co-doping with both DP and VA was shown to significantly improve the electroactivity of PEDOT in comparison the individually doped coatings. Similarly, while the individually doped PEDOT coatings were mechanically friable, the inclusion of both dopants during electropolymerisation was shown to attenuate this response. In a whole-blood model of inflammation all DP and VA doped CPs retained their bioactivity, causing a significant reduction in levels of the pro-inflammatory cytokine TNF-α. These studies demonstrated that small charged bioactive molecules are able act as dopants for CPs and that co-doping with ions of varied size and doping affinity may provide a means of addressing the limitations of large bulky bimolecular dopants.


Confocal flowmeter based on self-mixing interferometry for real-time velocity profiling of turbid liquids flowing in microcapillaries.
We propose a new confocal device for flow profiling in microcapillaries. A viewfinder system is developed using a visible light microscope, allowing focusing with high precision an 830 nm Fabry-Perot laser diode on a microchannel. By means of a novel confocal approach, the Doppler shift produced by the particles of a turbid liquid moving in the focal plane can be measured in real time using the well-known self-mixing effect. The resolution of this device is characterized in function of the full width at half maximum of the Gaussian frequency peak related to the self-mixing signal in the frequency domain. Velocity measurements for flow rates from 0.2 to 1.6 mL/min are presented, and the results demonstrate that the method reduces the phase noise and the effects of the out-of-focus particles, allowing straightforward flow profiling in microchannel structures.


The Arabidopsis ATR1 Myb transcription factor controls indolic glucosinolate homeostasis.
Plants derive a number of important secondary metabolites from the amino acid tryptophan (Trp), including the growth regulator indole-3-acetic acid (IAA) and defense compounds against pathogens and herbivores. In previous work, we found that a dominant overexpression allele of the Arabidopsis (Arabidopsis thaliana) Myb transcription factor ATR1, atr1D, activates expression of a Trp synthesis gene as well as the Trp-metabolizing genes CYP79B2, CYP79B3, and CYP83B1, which encode enzymes implicated in production of IAA and indolic glucosinolate (IG) antiherbivore compounds. Here, we show that ATR1 overexpression confers elevated levels of IAA and IGs. In addition, we show that an atr1 loss-of-function mutation impairs expression of IG synthesis genes and confers reduced IG levels. Furthermore, the atr1-defective mutation suppresses Trp gene dysregulation in a cyp83B1 mutant background. Together, this work implicates ATR1 as a key homeostatic regulator of Trp metabolism and suggests that ATR1 can be manipulated to coordinately control the suite of enzymes that synthesize IGs.


Moringa oleifera Lam and its Therapeutic Effects in Immune Disorders.
Moringa oleifera Lam., a plant native to tropical forests of India, is characterized by its versatile application as a food additive and supplement therapy. Accumulating evidence shows that Moringa plays a critical role in immune-related diseases. In this review, we cover the history, constituents, edibility, and general medicinal value of Moringa. The effects of Moringa in treating immune disorders are discussed in detail. Moringa can not only eliminate pathogens, including bacteria, fungi, viruses, and parasites, but also inhibit chronic inflammation, such as asthma, ulcerative colitis, and metabolic diseases. Additionally, Moringa can attenuate physical and chemical irritation-induced immune disorders, such as metal intoxication, drug side effects, or even the adverse effect of food additives. Autoimmune diseases, like rheumatoid arthritis, atopic dermatitis, and multiple sclerosis, can also be inhibited by Moringa. Collectively, Moringa, with its multiple immune regulatory bioactivities and few side effects, has a marked potential to treat immune disorders.


Characterization of microRNA expression in primary human colon adenocarcinoma cells (SW480) and their lymph node metastatic derivatives (SW620).
Metastasis is the major cause of cancer-related deaths in patients with colon cancer, however, the exact molecular mechanism is unclear. MicroRNAs (miRNAs) play an important role in the pathogenesis and progression of cancer. Therefore, in this study, we aimed to identify differentially expressed miRNAs in two colon carcinoma cell lines: SW480, derived from primary colon carcinoma and SW620, derived from lymph node metastasis, which were obtained from the same patient. Three independent samples of cancer cells were collected from SW480 and SW620 cells, respectively. An miRNA microarray platform, miRCURY LNA™ microRNA array with 1,223 probes containing 3,000 capture probes, was used to determine the miRNA expression profiles of these two cell lines. Differentially expressed miRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The raw data were submitted to the Gene Expression Omnibus database (GSE72412). Thirteen miRNAs were differentially expressed between SW480 and SW620 cells, of which, seven miRNAs (hsa-miR-920, hsa-miR-636, hsa-miR-766-3p, hsa-miR-545-5p, hsa-miR-195-3p, hsa-miR-125a-3p, and hsa-miR-196b-3p) were found to be upregulated and six miRNAs (hsa-miR-3613-3p, hsa-miR-29b-3p, hsa-miR-1297, hsa-miR-141-5p, hsa-miR-200c-3p, and hsa-miR-141-3p) were found to be downregulated. Target analysis of the predicted miRNAs showed that these genes were primarily involved in protein binding, cell adhesion, and cancer metastasis. Furthermore, qRT-PCR validated the results of miRNA microarray. This is the first systematic analysis of the differences of miRNAs between SW480 and SW620 cells. The results provide useful information to explore potential biomarkers of miRNAs for predicting colon cancer metastasis.


Efficacy and safety of pimecrolimus cream in the long-term management of atopic dermatitis in children.
Pimecrolimus cream (SDZ ASM 981), a nonsteroid inhibitor of inflammatory cytokines, is effective in atopic dermatitis (AD). We assessed whether early treatment of AD signs/symptoms with pimecrolimus could influence long-term outcome by preventing disease flares. Early intervention with pimecrolimus was compared with a conventional AD treatment strategy (ie, emollients and topical corticosteroids). In this 1-year, controlled, double-blind study, 713 AD patients (2-17 years) were randomized 2:1 to a pimecrolimus-based or conventional regimen. Both groups used emollients for dry skin. Early AD signs/symptoms were treated with pimecrolimus cream or, in the conventional treatment group, vehicle to prevent progression to flares. If flares occurred, moderately potent topical corticosteroids were mandated. The primary efficacy endpoint was ranked flares at 6 months. Safety was monitored clinically, and a skin recall-antigen test was performed at study completion. BASELINE CHARACTERISTICS OF THE PATIENTS: The mean age for both groups was approximately 8 years, and the majority of patients had moderate disease at baseline. PATIENT FOLLOW-UP AND EXPOSURE TO STUDY MEDICATION: The mean duration of follow-up (+/-standard error) was 303.7 (+/-5.30) days in the pimecrolimus group and 235.2 (+/-9.40) days in the control group. The discontinuation rate was significantly higher in the control group than in the pimecrolimus group (51.5% vs 31.6% at 12 months), and proportionately more patients with severe or very severe disease discontinued in the control group. The main reason for the higher discontinuation rate in the control group was unsatisfactory therapeutic effect (30.4% vs 12.4%). This resulted in a substantially higher mean number of study medication treatment days in the pimecrolimus group compared with the control group: 211.9 (69.8% of study days) versus 156.0 (66.3% of study days). Of those patients who completed 12 months on study, 14.2% and 7.0% of patients in the pimecrolimus and vehicle groups, respectively, used study medication continuously. Patients in the pimecrolimus group experienced significantly fewer AD flares than those in the control group, according to the primary efficacy analysis on ranked flares of AD (Van Elteren test). The proportion of patients who completed 6 or 12 months with no flares was approximately twice as high in the pimecrolimus group compared with control (61.0% vs 34.2% at 6 months; 50.8% vs 28.3% at 12 months). Fewer flares were observed in the pimecrolimus group regardless of baseline disease severity, so even severe patients derived benefit from the treatment. The analysis of time to first flare showed that treatment with pimecrolimus was associated with a significantly longer flare-free period (log- rank test). Covariate analysis indicated a statistically significant effect on time to first flare of baseline Eczema Area and Severity Index score, and whether patients had "severe" or "very severe" disease at baseline according to the Investigators' Global Assessment, although patients in all baseline disease severity subgroups benefited from treatment. Age had no significant effect. Fewer patients in the pimecrolimus group required topical corticosteroid therapy compared with control (35.0% vs 62.9% at 6 months; 42.6% vs 68.4% at 12 months), and patients in the pimecrolimus group spent fewer days on topical corticosteroid therapy (57.4% vs 31.6% [pimecrolimus vs control, respectively] spent 0 days on topical corticosteroid therapy, 17.1% vs 27.5% 1-14 days, and 25.5% vs 41.0% >14 days over the 12 months of the study). This steroid-sparing effect of pimecrolimus was evident despite pimecrolimus-treated patients being on study longer than patients in the control group. The average proportion of study days spent on second-line corticosteroids was 4.08% in the pimecrolimus group and 9.10% in the control group. Analysis of Eczema Area and Severity Index over time showed significantly lower median scores, thus indicating better disease control in the pimecrolimus group compared with the control group. Similar results were obtained from analysis of the Investigators' Global Assessment (not shown). The treatment groups were well balanced with respect to the number of patients using antihistamines during the study (57.2% vs 62.9%, pimecrolimus vs control, respectively). There were no appreciable differences between treatment groups in the overall incidence of adverse events. The most frequent adverse events were common childhood infections and ailments, including nasopharyngitis, headache, and cough. The incidence of suspected drug-related adverse events was not significantly different in the pimecrolimus group (24.7% vs 18.7%--pimecrolimus vs control), and the incidence of serious adverse events was low (8.3% vs 5.2%--pimecrolimus vs control). Life-table analysis of incidence of adverse events revealed no significant differences between the treatment groups, except for cough. Local tolerability was good in both treatment groups. The most common application site reaction reported was sensation of burning (10.5% vs 9.3%--pimecrolimus vs control). There were no major differences between treatment groups in the duration or severity of application site reactions, most of which were mild-to-moderate and transient, occurring within the first week of treatment. Skin infections were reported in both groups. There were no between-group differences in the life-table analysis of time to first occurrence of bacterial skin infections nor in the adjusted incidence of bacterial skin infections. Although there were no significant differences between treatment groups in the incidence of individual viral skin infections, the incidence of grouped viral skin infections (12.4% vs 6.3%--pimecrolimus vs control) showed a slightly higher incidence in the pimecrolimus group. Laboratory values and vital signs showed no significant between-group differences. There were no significant differences between treatment groups in response to recall antigens in those patients who remained on study for 12 months. Treatment of early AD signs/symptoms with pimecrolimus was effective in preventing progression to flares in more than half the patients, reducing or eliminating the need for topical corticosteroids. The benefits were consistently seen at 6 months across important disease severity subgroups and with respect to the various predefined efficacy endpoints. Furthermore, these benefits were sustained for 12 months, providing evidence that long-term treatment with pimecrolimus leads to better control of AD. Treatment with pimecrolimus was well tolerated and was not associated with clinically relevant adverse events compared with the conventional treatment group. The results reported here offer the prospect of effective long-term management of AD with reduced need for topical corticosteroids.


Evidence for corticotropin-releasing factor, neurotensin, and somatostatin in the neural pathway from the central nucleus of the amygdala to the parabrachial nucleus.
The central nucleus of the amygdala (CNA) and the parabrachial nucleus of the pons (PBN) are included within a group of brain nuclei involved in autonomic responses. Previous studies have shown that the CNA sends a considerable projection to the PBN and that both nuclei contain neurons immunoreactive to many different peptides. In the present study, we used the combined retrograde fluorescence-immunofluorescence method to determine whether the CNA projection to the PBN contains any of the following neuropeptides: corticotropin-releasing factor (CRF), neurotensin (NT), somatostatin (SS), and enkephalin (ENK). Following injections of fluorescent dye into the PBN, neurons within both lateral and medial subdivisions of the CNA were retrogradely labeled. A significant percentage of CRF (54-66%)-, NT (40-53%)-, and SS (31-50%)-immunoreactive neurons were retrogradely labeled, predominantly within the lateral CNA. Enkephalin-immunoreactive neurons were never retrogradely labeled, although they were often found adjacent to retrogradely labeled neurons. Our results show that the lateral CNA is a major source of CRF, NT, and SS terminals within the PBN. Neurons in the medial CNA also provide a significant contribution to the CNA-PBN pathway, but their chemical nature remains to be determined. We conclude that CRF, NT, and SS are important putative neurotransmitters in the CNA's regulation of PBN function. This CNA-PBN peptidergic pathway may participate in stress-related cardiovascular and respiratory responses.


Physical activity in adults with cystic fibrosis receiving intravenous antibiotics in hospital and in the community.
Intravenous antibiotic therapy (IVAT) for CF acute pulmonary exacerbations (APE) can be delivered in hospital or in the community. This study aimed to compare physical activity in CF patients receiving hospital and community-delivered IVAT, as well as other health outcomes. This was a non-randomised parallel group prospective observational study. Hospitalised and community-treated CF adults receiving IVAT for APE were asked to wear ActiGraph® activity monitors, complete the habitual activity estimation scale (HAES), food diary, modified shuttle test (MST) and CFQ-R at the start and end of therapy. Nutritional and clinical outcomes were also compared between the cohorts. The primary outcomes was physical activity measured by the ActiGraph® activity monitors at the beginning and end of treatment in both cohorts. Physical activity (measured and self-reported) was no different between the cohorts, with both hospitalised and community-treated subjects being generally sedentary. Body weight increased significantly in the hospitalised cohort, whereas no difference was seen in the community-treated cohort. FEV1 % predicted and FVC % predicted increased in community-treated subjects, whereas only FVC % predicted increased in hospitalised subjects. CFQ-R respiratory domain increased to a greater extent in community-treated subjects. CF adults receiving IVAT for APE, both in hospital and in the community, are generally sedentary and we found no difference in physical activity between the two groups. These findings suggests the need to further promote physical activity in suitable patients during APE where considered appropriate.


Plasmonic Bubble Nucleation and Growth in Water: Effect of Dissolved Air.
Under continuous laser irradiation, noble metal nanoparticles immersed in water can quickly heat up, leading to the nucleation of so-called plasmonic bubbles. In this work, we want to further understand the bubble nucleation and growth mechanism. In particular, we quantitatively study the effect of the amount of dissolved air on the bubble nucleation and growth dynamics, both for the initial giant bubble, which forms shortly after switching on the laser and is mainly composed of vapor, and for the final life phase of the bubble, during which it mainly contains air expelled from water. We found that the bubble nucleation temperature depends on the gas concentration: the higher the gas concentration, the lower the bubble nucleation temperature. Also, the long-term diffusion-dominated bubble growth is governed by the gas concentration. The radius of the bubbles grows as R(t) ∝ t 1/3 for air-equilibrated and air-oversaturated water. In contrast, in partially degassed water, the growth is much slower since, even for the highest temperature we achieve, the water remains undersaturated.


Licochalcone-E induces caspase-dependent death of human pharyngeal squamous carcinoma cells through the extrinsic and intrinsic apoptotic signaling pathways.
The aim of the present study was to investigate licochalcone-E (Lico-E)-induced apoptosis and the associated apoptotic signaling pathway in FaDu cells, a human pharyngeal squamous carcinoma cell line. Treatment with Lico-E exhibited significant cytotoxicity on FaDu cells in a concentration-dependent manner. The IC50 value of Lico-E in FaDu cells was ~50 µM. Treatment with Lico-E increased the number of dead FaDu cells. Furthermore, chromatin condensation, which is associated with apoptotic cell death, was observed in FaDu cells treated with Lico-E for 24 h. By contrast, Lico-E did not produce cytotoxicity or increase the number of dead cells when applied to human normal oral keratinocytes (hNOKs). Furthermore, chromatin condensation was not observed in hNOKs treated with Lico-E. Treatment with Lico-E increased the expression of Fas ligand and the cleaved form of caspase-8 in FaDu cells. Furthermore, treatment with Lico-E increased the expression of pro-apoptotic factors, including apoptosis regulator BAX, Bcl-2-associated agonist of cell death, apoptotic protease-activating factor 1, caspase-9 and tumor suppressor p53, while decreasing the expression of anti-apoptotic factors, including apoptosis regulator Bcl-2 and Bcl-2-like protein 1 in FaDu cells. The expression of cleaved caspases-3 and poly (ADP-ribose) polymerase was significantly upregulated following treatment with Lico-E in FaDu cells, while Lico-E-induced apoptotic FaDu cell death was partially suppressed by treatment with Z-VAD-FMK, a pan caspase inhibitor. Therefore, Lico-E-induced oral cancer (OC) cell-specific apoptosis is mediated by the death receptor-dependent extrinsic and mitochondrial-dependent intrinsic apoptotic signaling pathways. In conclusion, these data suggested that Lico-E exhibits potential chemopreventive effects and warrants further developed as a chemotherapeutic agent against OC.


Interventions to improve continuity of care in the follow-up of patients with cancer.
Care from the family physician is generally interrupted when patients with cancer come under the care of second-line and third-line healthcare professionals who may also manage the patient's comorbid conditions. This situation may lead to fragmented and uncoordinated care, and results in an increased likelihood of not receiving recommended preventive services or recommended care. To classify, describe and evaluate the effectiveness of interventions aiming to improve continuity of cancer care on patient, healthcare provider and process outcomes. We searched the Cochrane Effective Practice and Organization of Care Group (EPOC) Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, EMBASE, CINAHL, and PsycINFO, using a strategy incorporating an EPOC Methodological filter. Reference lists of the included study reports and relevant reviews were also scanned, and ISI Web of Science and Google Scholar were used to identify relevant reports having cited the studies included in this review. Randomised controlled trials (including cluster trials), controlled clinical trials, controlled before and after studies and interrupted time series evaluating interventions to improve continuity of cancer care were considered for inclusion. We included studies that involved a majority (> 50%) of adults with cancer or healthcare providers of adults with cancer. Primary outcomes considered for inclusion were the processes of healthcare services, objectively measured healthcare professional, informal carer and patient outcomes, and self-reported measures performed with scales deemed valid and reliable. Healthcare professional satisfaction was included as a secondary outcome. Two reviewers described the interventions, extracted data and assessed risk of bias. The authors contacted several investigators to obtain missing information. Interventions were regrouped by type of continuity targeted, model of care or interventional strategy and were compared to usual care. Given the expected clinical and methodological diversity, median changes in outcomes (and bootstrap confidence intervals) among groups of studies that shared specific features of interest were chosen to analyse the effectiveness of included interventions. Fifty-one studies were included. They used three different models, namely case management, shared care, and interdisciplinary teams. Six additional interventional strategies were used besides these models: (1) patient-held record, (2) telephone follow-up, (3) communication and case discussion between distant healthcare professionals, (4) change in medical record system, (5) care protocols, directives and guidelines, and (6) coordination of assessments and treatment.Based on the median effect size estimates, no significant difference in patient health-related outcomes was found between patients assigned to interventions and those assigned to usual care. A limited number of studies reported psychological health, satisfaction of providers, or process of care measures. However, they could not be regrouped to calculate median effect size estimates because of a high heterogeneity among studies. Results from this Cochrane review do not allow us to conclude on the effectiveness of included interventions to improve continuity of care on patient, healthcare provider or process of care outcomes. Future research should evaluate interventions that target an improvement in continuity as their primary objective and describe these interventions with the categories proposed in this review. Also of importance, continuity measures should be validated with persons with cancer who have been followed in various settings.


Root vacuolar Na+ sequestration but not exclusion from uptake correlates with barley salt tolerance.
Soil salinity is a major constraint for the global agricultural production. For many decades, Na+ exclusion from uptake has been the key trait targeted in breeding programs; yet, no major breakthrough in creating salt-tolerant germplasm was achieved. In this work, we have combined the microelectrode ion flux estimation (MIFE) technique for non-invasive ion flux measurements with confocal fluorescence dye imaging technique to screen 45 accessions of barley to reveal the relative contribution of Na+ exclusion from the cytosol to the apoplast and its vacuolar sequestration in the root apex, for the overall salinity stress tolerance. We show that Na+ /H+ antiporter-mediated Na+ extrusion from the root plays a minor role in the overall salt tolerance in barley. At the same time, a strong and positive correlation was found between root vacuolar Na+ sequestration ability and the overall salt tolerance. The inability of salt-sensitive genotypes to sequester Na+ in root vacuoles was in contrast to significantly higher expression levels of both HvNHX1 tonoplast Na+ /H+ antiporters and HvVP1 H+ -pumps compared with tolerant genotypes. These data are interpreted as a failure of sensitive varieties to prevent Na+ back-leak into the cytosol and existence of a futile Na+ cycle at the tonoplast. Taken together, our results demonstrated that root vacuolar Na+ sequestration but not exclusion from uptake played the main role in barley salinity tolerance, and suggested that the focus of the breeding programs should be shifted from targeting genes mediating Na+ exclusion from uptake by roots to more efficient root vacuolar Na+ sequestration.


The Opitz syndrome gene product, MID1, associates with microtubules.
Opitz syndrome (OS) is a genetically heterogeneous disorder characterized by defects of the ventral midline, including hypertelorism, cleft lip and palate, heart defects, and mental retardation. We recently identified the gene responsible for X-linked OS. The ubiquitously expressed gene product, MID1, is a member of the RING finger family. These proteins are characterized by an N-terminal tripartite protein-protein interaction domain and a conserved C terminus of unknown function. Unlike other RING finger proteins for which diverse cellular functions have been proposed, the function of MID1 is as yet undefined. By using the green fluorescent protein as a tag, we show here that MID1 is a microtubule-associated protein that influences microtubule dynamics in MID1-overexpressing cells. We confirm this observation by demonstrating a colocalization of MID1 and tubulin in subcellular fractions and the association of endogenous MID1 with microtubules after in vitro assembly. Furthermore, overexpressed MID1 proteins harboring mutations described in OS patients lack the capability to associate with microtubules, forming cytoplasmic clumps instead. These data give an idea of the possible molecular pathomechanism underlying the OS phenotype.


[Genistein: a soy isoflavone revealing a pleiotropic mechanism of action - clinical implications in the treatment and prevention of cancer].
Genistein, a naturally occurring isoflavonoid, displays antitumor, antimetastatic and antiangiogenic properties, described in various experimental in vitro and in vivo models. The results of several epidemiological studies suggest that soybean consumption may contribute to lower incidence of breast, colon, prostate, thyroid, and head and neck cancers. This protective effect of soy consumption is attributed, among others, to genistein. On the other hand, genistein may enhance the proliferation of some estrogen-positive human breast cancer cells in vivo and the growth of mammary gland and mammary cancer cells in athymic mice. In this paper, various aspects of the diverse biological activities of genistein and their potential clinical implications, especially in the treatment and prevention of cancer, are reviewed and discussed.


Association of Changes in Cardiovascular Health Metrics and Risk of Subsequent Cardiovascular Disease and Mortality.
Background The extent to which change in cardiovascular health (CVH) in midlife reduces risk of subsequent cardiovascular disease and mortality is unclear. Methods and Results CVH was computed at 2 ARIC (Atherosclerosis Risk in Communities) study visits in 1987 to 1989 and 1993 to 1995, using 7 metrics (smoking, body mass index, total cholesterol, blood glucose, blood pressure, physical activity, and diet), each classified as poor, intermediate, and ideal. Overall CVH was classified as poor, intermediate, and ideal to correspond to 0 to 2, 3 to 4, and 5 to 7 metrics at ideal levels. There 10 038 participants, aged 44 to 66 years that were eligible. From the first to the second study visit, there was an improvement in overall CVH for 17% of participants and a decrease in CVH for 21% of participants. At both study visits, 28%, 27%, and 6% had poor, intermediate, and ideal overall CVH, respectively. Compared with those with poor CVH at both visits, the risk of cardiovascular disease (hazard ratio [HR], 0.26; 95% CI, 0.20-0.34) and mortality (HR, 0.35; 95% CI, 0.29-0.44) was lowest in those with ideal CVH at both measures. Improvement from poor to intermediate/ideal CVH was also associated with a lower risk of cardiovascular disease (HR, 0.67; 95% CI, 0.59-0.75) and mortality (HR, 0.80; 95% CI, 0.72-0.89). Conclusions Improvement in CVH or stable ideal CVH, compared with those with poor CVH over time, is associated with a lower risk of incident cardiovascular disease and all-cause mortality. The change in smoking status and cholesterol may have accounted for a large part of the observed association.


U6 RNA biogenesis and disease association.
U6 snRNA is one of five uridine-rich noncoding RNAs that form the major spliceosome complex. Unlike other U-snRNAs, it reveals many distinctive aspects of biogenesis such as transcription by RNA polymerase III, transcript nuclear retention and particular features of transcript ends: monomethylated 5'-guanosine triphosphate as cap structure and a 2',3'-cyclic phosphate moiety (>P) at the 3' termini. U6-snRNA plays a central role in splicing and thus its transcription, maturation, snRNP formation, and recycling are essential for cellular homeostasis. U6 snRNA enters the splicing cycle as part of the tri-U4/U6.U5snRNP complex, and after significant structural arrangements forms the catalytic site of the spliceosome together with U2 snRNA and Prp8. U6 snRNA also contributes to the splicing reaction by coordinating metal cations required for catalysis. Many human diseases are associated with altered splicing processes. Disruptions of the basal splicing machinery can be lethal or lead to severe diseases such as spinal muscular atrophy, amyotrophic lateral sclerosis, or retinitis pigmentosa. Recent studies have identified a new U6 snRNA biogenesis factor Usb1, the absence of which leads to poikiloderma with neutropenia (PN) (OMIM 604173), an autosomal recessive skin disease. Usb1 is an evolutionarily conserved 3'→5' exoribonuclease that is responsible for removing 3'-terminal uridines from U6 snRNA transcripts, which leads to the formation of a 2',3' cyclic phosphate moiety (>P). This maturation step is fundamental for U6 snRNP assembly and recycling. Usb1 represents the first example of a direct association between a spliceosomal U6 snRNA biogenesis factor and human genetic disease.


[Malignant fibrous histiocytoma of the right ventricle and pulmonary artery. Apropos of a case. Review of the literature].
The authors report a case of malignant cardiac fibrohistiocytoma involving the right ventricular infundibulum and the pulmonary artery and responsible for a pseudo-thromboembolic right heart failure syndrome. Following discovery of the tumour mass on ultrasonography and catheterisation, wide excision of the pulmonary artery, pulmonary valve and the roof of the infundibulum as far as the middle part of the ventricle was performed under cardio-pulmonary by-pass. Reconstruction was performed with a valveless woven Dacron tube. The postoperative course was uncomplicated and the patient was still alive after 18 months. Histology revealed malignant fibrohistiocytoma. The authors also present a review of the literature of these rare tumours of the right-sided cavities (5 cases reported).


Influence of auto-antibody specificities on the clinical course in patients with chronic and acute ITP.
It is now well established that the target antigens of platelet auto-antibodies are the GP IIb/IIIa and GP Ib/IX receptors. In our study, it was observed that the majority of patients had antibodies directed towards the GP IIb/IIIa receptor, whereas the number of patients having antibodies directed towards the GP Ib/IX receptor was much less. Also, we found that the patients in whom the auto-antibodies are directed towards the GP IIb/IIIa receptors usually have mild bleeding. On the other hand, the patients having auto-antibodies directed towards both GP IIb/IIIa and GP Ib/IX receptors have a severe bleeding diathesis, and usually show poor prognosis to treatment with corticosteroids.


Guanidine transport across the apical and basolateral membranes of human intestinal Caco-2 cells is mediated by two different mechanisms.
The functional characteristics of the intestinal absorption and secretion of guanidine as a model of a nutritionally and metabolically essential organic cation were examined in the Caco-2 human intestinal cell line. Both apical and basolateral transport of [14C]-guanidine were studied using Caco-2 cells grown on polycarbonate permeable membranes. The basolateral-to-apical flux of [14C]-guanidine (i.e., its secretion) was quantitatively higher than the apical-to-basolateral transport (i.e., its absorption). When Na+ was replaced by K+ or Li+, both apical and basolateral accumulation were significantly inhibited. Studies using the cell monolayers and apical membrane vesicles obtained from Caco-2 cells showed a potential-independent mechanism of guanidine apical uptake and efflux. Conversely, basolateral uptake and efflux were membrane potential dependent. Kinetic analysis revealed that both saturable and nonsaturable mechanisms accounted for the apical and basolateral accumulations. The [14C]-guanidine efflux from cells through the apical and basolateral membranes was significantly reduced at 4 degrees C, suggesting carrier-mediated mechanisms. Moreover, the apical efflux was stimulated by an inwardly directed H+ gradient. Influx and efflux of [14C]-guanidine were unaffected by the presence of tetraethylammonium, cimetidine or decynium-22 in the donor compartment. Only quinine significantly reduced [14C]-guanidine entrance through apical and basolateral membranes and its exit through the basolateral membrane. In conclusion, our results suggest that the influx and the efflux through the apical membrane is mediated by different transporters, whereas transport across the basolateral membrane is mediated by a member of the organic cation transporter family with high affinity for guanidine.


Estimation of illumination characteristics.
The description of the relation between the one-parameter subgroups of a group and the differential operators in the Lie-algebra of the group is one of the major topics in Lie-theory. We use this framework to derive a partial differential equation which describes the relation between the time-change of the spectral characteristics of the illumination source and the change of the color pixels in an image. We introduce and justify the usage of conical coordinate systems in color space. We also derive the differential equation describing the illumination change and illustrate the algorithm with some simulation examples.


Generation of optical vortices with an adaptive helical mirror.
Generation of optical vortices using a new design of adaptive helical mirror (AHM) is reported. The new AHM is a reflective device that can generate an optical vortex of any desired topological charge, both positive and negative, within its breakdown limits. The most fascinating feature of the AHM is that the topological charge of the optical vortex generated with it can be changed in real time by varying the excitation voltage. Generation of optical vortices up to topological charge 4 has been demonstrated. The presence of a vortex in the optical field generated with the AHM is confirmed by producing both fork and spiral fringes in an interferometric setup. Various design improvements to further enhance the performance of the reported AHM are discussed. Some of the important applications of AHM are also listed.


Effects of nitric oxide on human spermatozoa activity, fertilization and mouse embryonic development.
This study was conducted to investigate the effects of nitric oxide (NO) on human sperm activity, human sperm-oocyte fusion and mouse embryonic development. Results showed that various concentrations of NO synthase blocker, N(omega)-nitro-L-arginine methyl ester, did not affect sperm cell motility at 0, 1, 2 or 4 hr, respectively. In contrast, sodium nitroprusside (SNP) significantly inhibited sperm cell motility and caused apoptosis. The adversely dose-dependent effect was only observed if SNP was freshly prepared. Adenosine triphosphate reversed the hazardous effect of SNP on sperm activity/viability. Hemoglobin neutralized the adverse effect of SNP. In hemi-zona sperm fusion test, the number of sperm bound to the zona in the presence of 10(-4) M SNP was significantly less than the control group. SNP at 10(-4) M caused all mouse embryonic development arrest. 46% and 56% of zygote reached the blastocyst stage with the treatment of 10(-6) M and 10(-8) M SNP, respectively, while the control reached 70%. NO adversely affected human sperm activity, human sperm-zona binding and embryonic development. It would appear that high concentration of NO may potentially decrease fertility.


Water recovery from sewage using forward osmosis.
This research is part of the Sewer Mining project aimed at developing a new technological concept by extracting water from sewage by means of forward osmosis (FO). FO, in combination with a reconcentration system, e.g. reverse osmosis (RO) is used to recover high-quality water. Furthermore, the subsequent concentrated sewage (containing an inherent energy content) can be converted into a renewable energy (RE) source (i.e. biogas). The effectiveness of FO membranes in the recovery of water from sewage has been evaluated. Stable FO water flux values (>4.3 LMH) were obtained with primary effluent (screened, not treated) used as the feed solution. Fouling of the membrane was also induced and further investigated. Accumulated fouling was found to be apparent, but not irreversible. Sewer Mining could lead to a more economical and sustainable treatment of wastewater, facilitating the extraction of water and energy from sewage and changing the way it is perceived: not as waste, but as a resource.


Stressful politics: voters' cortisol responses to the outcome of the 2008 United States Presidential election.
Social subordination can be biologically stressful; when mammals lose dominance contests they have acute increases in the stress hormone cortisol. However, human studies of the effect of dominance contest outcomes on cortisol changes have had inconsistent results. Moreover, human studies have been limited to face-to-face competitions and have heretofore never examined cortisol responses to shifts in political dominance hierarchies. The present study investigated voters' cortisol responses to the outcome of the 2008 United States Presidential election. 183 participants at two research sites (Michigan and North Carolina) provided saliva samples at several time points before and after the announcement of the winner on Election Night. Radioimmunoassay was used to measure levels of cortisol in the saliva samples. In North Carolina, John McCain voters (losers) had increases in post-outcome cortisol levels, whereas Barack Obama voters (winners) had stable post-outcome cortisol levels. The present research provides novel evidence that societal shifts in political dominance can impact biological stress responses in voters whose political party becomes socio-politically subordinate.


Evaluation of Fever in a child aged three months to 24 months.
Fever in the young infant may be especially challenging to the attending physician, particularly when no focus for the infection can be found on physical examination. Can the physician's clinical judgement be trusted? What constitutes a fever? How was the temperature measured? Can the fever harm the child? Does the child have a benign viremia or does the fever indicate a more serious underlying bacterial illness? Does a blood culture, a spinal tap, or a complete blood count need to be done? Should antibiotics or just antipyretics be prescribed? Should antipyretics be used in all febrile children; if so, in what dose? These questions have been considered at one time or another by all physicians taking care of children. Only with a comprehensive, scientific, and practical approach can the febrile infant be optimally managed.


Gender perspective in the analysis of the relationship between long workhours, health and health-related behavior.
The objective of this study was to analyze gender differences in the impact of long workhours (>40 hours per week) on a variety of health outcomes and health-related behavior. The sample included all salaried contract workers aged 16-64 years (1658 men and 1134 women) and interviewed in the 2002 Catalonian Health Survey. Whereas the men with a high job status were more likely to work >40 hours a week, long workhours were associated with situations of vulnerability (low job status and being separated or divorced) among the women. For both genders, working >40 hours was related to a shortage of sleep [adjusted odds ratio (aOR) 1.54, 95% confidence interval (95% CI) 1.21-1.98, for the men and aOR 1.63, 95% CI 1.11-2.38, for the women]. Among the women, long workhours were also associated with poor mental health status (aOR 1.58, 95% CI 1.04-2.40), hypertension (aOR 2.25, 95% CI 1.17-4.32), job dissatisfaction (aOR 1.77, 95% CI 1.08-2.90), and smoking (aOR 1.71, 95% CI 1.22-2.39). In addition, among the women working more hours at home, long workhours were related to sedentary leisure time activity (aOR 1.98, 95% CI 1.06-3.71). The relationship between long workhours and health and health-related behavior was found to be directly related to long worktime and indirectly related to long exposure to poor work conditions among the women and, to a less extent, to domestic work. The pathways that explain the relationship between long workhours and health and health-related behavior seems to depend on the outcome being analyzed.


FAS/FAE: impact on children.
Alcohol is a teratogen and it can cause lasting birth defects called Fetal Alcohol Syndrome (FAS)/Fetal Alcohol Effects (FAE). FAS is one of the leading causes of mental retardation. FAS/FAE can result in a myriad of behavioral, learning and developmental problems. There are estimates of 12,000 new cases of FAS yearly. The school nurse plays an important advocacy and educational role in helping the child with FAS. Family involvement is important for the child with FAS.