Combinatorial genetics in liver repopulation and carcinogenesis with a in vivo CRISPR activation platform.
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 activation (CRISPRa) systems have enabled genetic screens in cultured cell lines to discover and characterize drivers and inhibitors of cancer cell growth. We adapted this system for use in vivo to assess whether modulating endogenous gene expression levels can result in functional outcomes in the native environment of the liver. We engineered the catalytically dead CRISPR-associated 9 (dCas9)-positive mouse, cyclization recombination-inducible (Cre) CRISPRa system for cell type-specific gene activation in vivo. We tested the capacity for genetic screening in live animals by applying CRISPRa in a clinically relevant model of liver injury and repopulation. We targeted promoters of interest in regenerating hepatocytes using multiple single guide RNAs (gRNAs), and employed high-throughput sequencing to assess enrichment of gRNA sequences during liver repopulation and to link specific gRNAs to the initiation of carcinogenesis. All components of the CRISPRa system were expressed in a cell type-specific manner and activated endogenous gene expression in vivo. Multiple gRNA cassettes targeting a proto-oncogene were significantly enriched following liver repopulation, indicative of enhanced division of cells expressing the proto-oncogene. Furthermore, hepatocellular carcinomas developed containing gRNAs that activated this oncogene, indicative of cancer initiation events. Also, we employed our system for combinatorial cancer genetics in vivo as we found that while clonal hepatocellular carcinomas were dependent on the presence of the oncogene-inducing gRNAs, they were depleted for multiple gRNAs activating tumor suppressors. The in vivo CRISPRa platform developed here allows for parallel and combinatorial genetic screens in live animals; this approach enables screening for drivers and suppressors of cell replication and tumor initiation. (Hepatology 2017).


Prosopis alba exudate gum as novel excipient for fish oil encapsulation in polyelectrolyte bead system.
In this work, a bottom-up approach based on the study of polyelectrolyte interactions was performed in order to evaluate the effect of Prosopis alba exudate gum as novel excipient for fish oil encapsulation in composed calcium-alginate-chitosan beads. Emulsion and beads properties such as oil distribution, encapsulation efficiency, yield, microstructure and thermo-oxidative protection were evaluated. Alginate and gum exert a synergistic effect on emulsion stability properties, which conducted to better oil distribution in the beads and higher encapsulation efficiencies (98%) and yield (89%). The positive effect of including the gum as wall material was observed in terms of a higher oil retention capacity of the alginate beads, improved oxidative thermal stability and better microstructural features. Present results are promising and allowed considering P. alba gum as a novel non-conventional polyelectrolyte for improving Ca-alginate beads microstructure and stability with the added benefit of taking advantage of an available resource currently untapped.


[Phagodeterrent and systemic activity of a fomulation derived from an extract of common rue (Ruta chalepensis, Rutaceae) on Hypsipyla grandella (Lepidoptera: Pyralidae) larvae].
A key neotropical pest of mahoganies (Swietenia spp.) and cedars (Cedrela spp.), the larva of Hypsipyla grandella (Zeller) commonly bores into the main shoot of these valuable timber species, causing forking of the stems. Therefore, it would be desirable to count upon a phagodeterrent and systemic product which could readily protect young tissues. Since crude extracts of common rue foliage (Ruta chalepensis L.) have phagodeterrent activity on H. grandella larva, a product was formulated by combining such an extract with a number of coadjuvants, and was assessed on 75-100 cm tall Spanish cedar (Cedrela odorata L.) trees, inside a greenhouse. A completely randomized design was used, with 10 trees per each one of the following treatments: the formulated product, an absolute control (distilled water), a relative control (carbofuran, 1%m/v in water), and the relative control without the crude extract. Number of attacks to the main shoot, fallen leaves and frass piles were recorded, as well as tunnel length. Regarding these variables, the formulated product provided excellent protection to treated trees, with only 0.1 attacks to the main shoot, on the average, at the end of the evaluation period. Likewise, when the systemic activity of the crude extract was assessed, substances present in it were able to translocate and get assimilated by the trees, thereafter causing phagodeterrence to larvae. Finally, an espectrophotometry analysis (performed at a wavelength of 355 nm) allowed us to determine the concentration (403+/-1 mg/l) and percentage (0.40%) of the flavonoid rutin in the crude extract, which eventually may be used as an analytical marker compound, should an industrial formulation be developed.


The new pentaborate Na3SrB5O10.
Sodium strontium pentaborate, Na(3)SrB(5)O(10), maintains the same, previously unobserved, structure type at 200, 250 and 293 K. The fundamental building units are anionic [B(5)O(10)](5-) groups distorted from mm2 point symmetry. The Sr atoms are eightfold coordinated by O atoms, forming trigonal dodecahedra. The Na atoms appear in three crystallographically different environments. The present single-crystal results correct a previous report in which a monoclinic cell was deduced for this compound on the basis of powder diffraction data. The structure of the title compound is discussed in the crystalochemical context of other borates with the same formula type. Although the unit cell of the present compound is similar to that determined in a previous study of the analogous Ca-containing compound, this study demonstrates that the structures of the two are different. These novel alkali-alkaline earth borates are considered as potential host materials for optical applications (fluorescence materials or phosphors).


Autophagy activation by rapamycin eliminates mouse Mallory-Denk bodies and blocks their proteasome inhibitor-mediated formation.
The proteasomal and lysosomal/autophagy pathways in the liver and other tissues are involved in several biological processes including the degradation of misfolded proteins. Exposure of hepatocyte cell lines to proteasome inhibitors (PIs) results in the formation of inclusions that resemble Mallory-Denk bodies (MDBs). Keratins are essential for MDB formation and keratin 8 (K8)-overexpressing transgenic mice are predisposed to MDB formation. We tested the hypothesis that PIs induce MDBs in vivo and that autophagy participates in MDB turnover. The effect of the PI bortezomib (which is used to treat some malignancies) on MDB formation was tested in K8-overexpressing mice and in cultured cells. Inclusion formation was examined using immune and conventional electron microscopy (EM). Bortezomib induced MDB-like inclusions composed of keratins, ubiquitin, and p62 in cultured cells. Short-term exposure to bortezomib induced similar inclusions in K8-overexpressing but not in nontransgenic mice, without causing liver injury. In bortezomib-treated mice, autophagy was activated in hepatocytes as determined by EM and biochemical analysis. Further activation of autophagy by rapamycin (Rap) decreased the number of inclusions in bortezomib-treated K8 transgenic mice significantly. Rap also led to resorption of spontaneously formed MDBs in aging K8-overexpressing mice. Immune EM demonstrated K8-positive and ubiquitin-positive structures in autophagic vacuoles in the mouse liver. PIs alone are sufficient to induce MDBs in susceptible animals, while Rap-mediated activation of autophagy prevents MDB formation and causes MDB resorption. These findings suggest that some patients treated with PIs may become predisposed to MDB formation. Autophagy provides a potential cellular mechanism for the resorption of cytoplasmic inclusions.


The variable clinical manifestations of ulnar neuropathies at the elbow.
In twenty-five cases of ulnar neuropathy at the elbow, the involvement of the fibres from three sensory and to four motor branches were examined clinically and, where possible, electrophysiologically. Of the sensory fibres, those from the terminal digital nerves were most commonly involved. The fibres to the hand muscles were much more frequently involved than those to the forearm muscles. These findings suggest that in ulnar neuropathies at the elbow there is variable damage to the fascicles within the nerve.


Reconstitution of beta-adrenergic receptors into phospholipid vesicles: restoration of [125I]iodohydroxybenzylpindolol binding to digitonin-solubilized receptors.
beta-adrenergic receptors were solubilized from rat erythrocyte plasma membranes using digitonin. Solubilized receptors were then reconstituted into phospholipid vesicles by the addition of dimyristoylphosphatidylcholine and removal of detergent. Vesicles were separated from residual soluble receptors and detergent by rate-zonal ultracentrifugation. Vesicles were monolamellar, 500-900 A in diameter, and had a lipid content of 6 mumol phospholipid/mg protein. Specific binding of the beta-adrenergic ligand [3H]dihydroalprenolol ([3H]DNA) was 0.9-1.9 pmol/mg protein. Reconstitution of receptors into vesicles restored their ability to bind [125I]iodohydroxybenzylpindolol ([125I]IHYP). This ligand does not bind to detergent-solubilized receptors. [125I]IHYP binding was saturable [Kd = 84 pM] and competed appropriately with (+) and (-) isomers of beta-adrenergic agonists and antagonists. These receptor vesicles therefore appear to be an excellent model system for the study of beta-adrenergic receptor function in a defined lipid milieu.


[Role of basolateral and apical respiration in the active transport of glucose and maltose in different segments of the small intestine under different functional conditions].
The active transport of free glucose and glucose released in membrane hydrolysis involves oxygenation of mucosal and seroseal surfaces as well as bilateral one. The glucose accumulation against the electrochemical gradient increases in transition from fasting to satiety. The data obtained suggest a hypothesis stating that enterocytes and, probably, other functionally polarized cells have not a single (basolateral) but double types of the cellular respiration: the apical as well as the basolateral ones. In addition to diffusion exchange of O2 and CO2 between the blood and the basolateral surface of cells, the gas exchange is supposed to occur at the apical surface as well. The O2 transport to this particular surface of cells can be based on the Na+ microcirculation. The interrelationship between the apical and basolateral types of respiration was shown to depend on the functional condition of the organism (satiety or hunger).


Salmonella enterica serovars from pigs on farms and after slaughter and validity of using bacteriologic data to define herd Salmonella status.
The primary objective of this study was to evaluate the validity of using data obtained from slaughtered pigs for farm-level epidemiologic studies of Salmonella. The study involved groups of pigs from five farms. Salmonella isolates were obtained from on-farm samples, and a total of 370 on-farm and an additional 486 isolates from samples collected after commercial slaughter were subsequently tested. Preharvest samples included feces of individual animals from defined groups of nursery and finishing pigs on commercial farms and swabs from trucks. Postslaughter samples were cecal contents and mesenteric lymph node samples. The concordance between Salmonella serovars isolated from on-farm samples and those serovars isolated after slaughter varied widely among farms. Results of paired lymph node and cecal cultures were strongly associated (odds ratio, 7.0), but the agreement between on-farm and postslaughter results at the pig level was poor (kappa = 0.34). The results support recent findings that risk of exposure to Salmonella during transport and lairage remains a concern under contemporary industry conditions. The findings further imply that slaughter plant studies based on phenotyping of Salmonella alone (such as serovars) may not reliably indicate the Salmonella status of commercial swine farms.


Clinical characteristics of HIV-infected patients with adjudicated heart failure.
Aims HIV-infected persons may have elevated risks for heart failure, but factors associated with heart failure in the modern era of HIV therapy are insufficiently understood. Despite substantial disagreement between physician-adjudicated heart failure and heart failure diagnosis codes, few studies of HIV cohorts have evaluated adjudicated heart failure. We evaluated associations of HIV viremia, immunosuppression, and cardiovascular risk factors with physician-adjudicated heart failure. Methods and results We analyzed clinical characteristics associated with heart failure in a cohort of 5041 HIV-infected patients receiving care at an urban hospital system between 2000 and 2016. We also evaluated characteristics of HIV-infected patients who screened negative for heart failure, screened positive for possible heart failure but did not have heart failure after adjudication, and had adjudicated heart failure. HIV-infected patients with heart failure ( N = 216) were older and more likely to be black, hypertensive, and have diabetes than HIV-infected patients without heart failure; heart failure with reduced ejection fraction was more common than heart failure with preserved ejection fraction. In our primary analyses restricted to HIV-infected patients whose heart failure diagnoses did not precede their HIV diagnoses ( N = 149), peak HIV viral load ≥100,000 copies/mL (odds ratio (OR) 2.12, 1.28-3.52) and nadir CD4 T-cell count <200 cells/mm3 (OR 2.35, 1.04-5.31) were associated with significantly elevated odds of heart failure. Overall, 30.6% of patients with any diagnosis code of heart failure had adjudicated heart failure. Conclusion Higher peak HIV viremia and lower CD4 cell nadir are associated with significantly elevated odds of heart failure for HIV-infected persons. Physician adjudication of heart failure may be helpful in HIV cohorts.


Applying Blockchain Technology to Enhance Clinical Trial Recruitment.
Patient recruitment for clinical trials is known to be a challenging aspect of clinical research. There are multiple competing concerns from the sponsor, patient and principal investigator's perspectives resulting in most clinical trials not meeting recruitment requirements on time. Conducting under-enrolled clinical trials affects the power of conclusive results or causes premature trial termination. The Blockchain is a distributed ledger technology originally applied in the financial sector. Its features as a peer-to-peer system with publicly audited transactions, data security, and patient privacy are a good fit for the needs of clinical trials recruitment. The "Smart Contract" is a programmable self-executing protocol that regulates the blockchain transactions. Given current recruitment challenges, we have proposed a blockchain model containing multiple trial-based contracts for trial management and patient engagement and a master smart contract for automated subject matching, patient recruitment, and trial-based contracts management.


Exploring communication apprehension in nursing and healthcare education: A scoping review.
Communication among healthcare professionals is critical to optimizing patient outcomes, and communication education is incorporated into the educational programs of healthcare professions students. Communication can be a source of stress for new healthcare professionals, however, there is a lack of research on communication apprehension (CA) in healthcare students, especially in nursing education. This scoping review sought to explore CA and factors associated with CA among healthcare professions students. This scoping review report was guided by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (Tricco et al., Annals of Internal Medicine, 2018, 169, 467-473). Twenty studies of healthcare professions students were included for analysis. The percentage of health professions students with high CA ranged from 0.7% to 27%. Five out of seven intervention studies demonstrated a significant decrease in CA after the intervention. This review identified inconsistent associations between CA and various individual and educational factors. More research is needed utilizing rigorous research designs, designs employing qualitative methodology, and studies to investigate the impact of language on CA in health professions students who speak English as a second language.


Polymer Stabilized Cholesteric Liquid Crystal Siloxane for Temperature-Responsive Photonic Coatings.
Temperature-responsive photonic coatings are appealing for a variety of applications, including smart windows. However, the fabrication of such reflective polymer coatings remains a challenge. In this work, we report the development of a temperature-responsive, infrared-reflective coating consisting of a polymer-stabilized cholesteric liquid crystal siloxane, applied by a simple bar coating method. First, a side-chain liquid crystal oligosiloxane containing acrylate, chiral and mesogenic moieties was successfully synthesized via multiple steps, including preparing precursors, hydrosilylation, deprotection, and esterification reactions. Products of all the steps were fully characterized revealing a chain extension during the deprotection step. Subsequently, the photonic coating was fabricated by bar-coating the cholesteric liquid crystal oligomer on glass, using a mediator liquid crystalline molecule. After the UV-curing and removal of the mediator, a transparent IR reflective polymer-stabilized cholesteric liquid crystal coating was obtained. Notably, this fully cured, partially crosslinked transparent polymer coating retained temperature responsiveness due to the presence of non-reactive liquid-crystal oligosiloxanes. Upon increasing the temperature from room temperature, the polymer-stabilized cholesteric liquid crystal coating showed a continuous blue-shift of the reflection band from 1400 nm to 800 nm, and the shift was fully reversible.


Five Strategies for Optimizing Instructional Materials: Instructor- and Learner-Managed Cognitive Load.
Researchers of cognitive load theory and the cognitive theory of multimedia learning have identified several strategies to optimize instructional materials. In this review article we focus on five of these strategies or solutions to problematic instructional designs in multimedia learning: (a) the multimedia principle (use visualizations and drawings to complement texts); (b) the split-attention effect or spatial contiguity principle (show texts contiguously or integrated with visualizations); (c) the redundancy effect, alike the coherence principle (remove nonessential learning information); (d) the signaling principle (cue or signal essential learning information); and (e) the transient information effect or segmenting principle (segment or control the pace of animations and videos). Usually, both cognitive theories have investigated solutions that instructors, teachers, and designers should pursue to optimize students' learning. Here, in a novel approach, we show that these strategies can also be used by learners who want to self-manage their cognitive load and learning process. We provide several examples of both instructor- and learner-managed solutions aligned with these strategies. When assessing which agent, either the instructor or the learner, was most effective, we observed mixed results in the literature. However, the expertise reversal effect may help predict the direction of these effects: novice students may learn better under instructor-managed conditions, whereas more expert students may learn more under learner-managed conditions.


[Analysis on the quality control of suspected occupational disease from the characteristics of applicants diagnosed with noise deafness].
Objective: To analyze the audiology and occupational health data of applicants diagnosed of occupational noise deafness, and to explore the influencing factors in the diagnosis of suspected occupational noise deafness. Methods: In May 2022, the information of patients diagnosed with occupational noise deafness in Peking University Third Hospital from January 2018 to December 2021 was collected, and the occupational health data of their working environment, clinical audiological examination results and diagnosis basis of occupational noise deafness were collected and analyzed. Multi-factor unconditional logistic regression analysis was used to analyze independent risk factors for the diagnosis of occupational noise deafness. Results: A total of 129 subjects were included, all of which were suspected cases of occupational noise deafness found in various occupational health examination institutions. Eight cases (6.20%) were diagnosed as occupational noise deafness, and 121 cases (93.80%) were non-occupational noise deafness. After hearing examination, only 27.27% (24/88) of the patients' audiological changes were consistent with the starting point of occupational noise deafness diagnosis. Further analysis of the noise intensity in the workplace showed that 16 patients were identified as non-occupational noise deafness because the noise intensity of the working environment was less than 85 dB. Logistic regression analysis showed that the working hours were more than 8 hours (OR=9.274, 95%CI: 1.388-61.950, P=0.022) and the noise intensity of the working environment (OR=1.189, 95%CI: 1.059-1.334, P=0.003) were independent risk factors for the diagnosis of occupational noise deafness. Conclusion: The exclusion rate of suspected occupational noise deafness found in occupational health examination is higher after adequate rest. The test results of working environment noise intensity provided by the employer can help to determine occupational noise deafness.


Combined resection of the liver and inferior vena cava for hepatic malignancy.
The objective of this paper is to review the results of combined resection of the liver and inferior vena cava for hepatic malignancy. The morbidity and mortality along with preliminary survival data are assessed in order to determine the utility of this aggressive approach to otherwise unresectable tumors. Involvement of the inferior vena cava has traditionally been considered a contraindication to resection for advanced tumors of the liver because the surgical risks are high and the long-term prognosis is poor. Progress in liver surgery allows resection in some cases. Twenty-two patients undergoing hepatic resection from 1997 to 2003, that also required resection and reconstruction of the inferior vena cava (IVC), were reviewed. The median age was 49 years (range 2 to 68 years). Resections were carried out for: hepatocellular carcinoma (n = 6), colorectal metastases (n = 6), cholangiocarcinoma (n = 5), gastrointestinal stromal tumor (n = 2), hepatoblastoma (n = 2), and squamous cell carcinoma in 1 patient. Liver resections performed included 13 right trisegmentectomies, 6 right lobectomies extended to include the caudate lobe, and 3 left trisegmentectomies. Complex ex vivo procedures were performed in 2 cases using venovenous bypass while the other 20 cases were performed using varying degrees of vascular isolation. In situ cold perfusion of the liver was used in 1 case. The IVC was reconstructed with ringed Gore-Tex tube graft (n = 14), primarily (n = 6), or with Gore-Tex patches (n = 2). There were 2 perioperative deaths (9%). One cirrhotic patient died of liver failure 3 weeks post operatively and 1 patient with cholangiocarcinoma died of pulmonary hemorrhage secondary to a cavitating pulmonary infection after aspiration pneumonia 6 weeks after resection. Six patients had evidence of postoperative liver failure that resolved with supportive management and 2 patients required temporary dialysis. All vascular reconstructions were patent at last follow-up. With median follow-up of 26 months, 5 patients have died of recurrent malignancy at 44, 40, 32, 26, and 24 months, while an additional patient is alive with disease at 31 months. Actuarial 1-, 3-, and 5-year survivals were 85%, 60%, and 33%, respectively. IVC involvement by hepatic malignancy does not necessarily preclude resection. Liver resection with reconstruction of the inferior vena cava can be performed in selected cases. The increased risk associated with the procedure appears to be balanced by the possible benefits, particularly when the lack of alternative curative approaches is considered.


The added value of orthotic management in the context of multi-level surgery in children with cerebral palsy.
Treatment of cerebral palsy includes an interdisciplinary concept and in more severe cases the well-established multi-level surgery (MLS). Different kinds of orthoses are typically part of postoperative treatment but there is a lack of knowledge about their additional benefit. Do ankle foot orthoses lead to an additional, measurable improvement of gait after MLS? 20 children with bilateral spastic cerebral palsy (9 retrospective, 11 in a postoperative clinical routine) were included. All had a preoperative gait analysis before MLS. Postoperatively, they were fitted with different ankle foot orthoses (AFO), depending on their individual needs. Dynamic ankle foot orthoses (DAFO), combined DAFO with additional dynamic, elastic shank adaptation (DESA) and ground reaction force AFOs (GRAFO) were used. Patients underwent a second gait analysis 1.5 (± 0.6) years postoperatively barefoot and with orthoses. Data analysis included testing for normal distribution (Shapiro-Wilk-Test) and further nonparametric statistical testing on basis of a Wilcoxon Single-Rank Test. The operation produced changes in the hip, knee and ankle joint, and the pelvis. Spatiotemporal parameters showed significant changes due to additional use of the orthoses. Further, additional kinematic changes occurred at the hip, knee and ankle joint as well as the foot. The Gillette Gait Index (GGI) improved significantly by supplementary orthoses, but not by surgery alone. The Gait Profile Score (GPS) and Gait Deviation Index (GDI) rather showed changes due to the surgery. MLS significantly improves GPS and GDI more than a year after surgery, which can be interpreted as an improvement in gait pattern. In contrast, the GGI is improved by additional postoperative orthotic treatment, which implies that walking ability itself has improved, rather than the gait pattern. Orthoses show a positive additional effect on surgical results at different anatomical levels. Spatiotemporal parameters are positively influenced solely by additional orthotic support.


Neonatal outcomes for women at risk of preterm delivery given half dose versus full dose of antenatal betamethasone: a randomised, multicentre, double-blind, placebo-controlled, non-inferiority trial.
Antenatal betamethasone is recommended before preterm delivery to accelerate fetal lung maturation. However, reports of growth and neurodevelopmental dose-related side-effects suggest that the current dose (12 mg plus 12 mg, 24 h apart) might be too high. We therefore investigated whether a half dose would be non-inferior to the current full dose for preventing respiratory distress syndrome. We designed a randomised, multicentre, double-blind, placebo-controlled, non-inferiority trial in 37 level 3 referral perinatal centres in France. Eligible participants were pregnant women aged 18 years or older with a singleton fetus at risk of preterm delivery and already treated with the first injection of antenatal betamethasone (11·4 mg) before 32 weeks' gestation. We used a computer-generated code producing permuted blocks of varying sizes to randomly assign (1:1) women to receive either a placebo (half-dose group) or a second 11·4 mg betamethasone injection (full-dose group) 24 h later. Randomisation was stratified by gestational age (before or after 28 weeks). Participants, clinicians, and study staff were masked to the treatment allocation. The primary outcome was the need for exogenous intratracheal surfactant within 48 h after birth. Non-inferiority would be shown if the higher limit of the 95% CI for the between-group difference between the half-dose and full-dose groups in the primary endpoint was less than 4 percentage points (corresponding to a maximum relative risk of 1·20). Four interim analyses monitoring the primary and the secondary safety outcomes were done during the study period, using a sequential data analysis method that provided futility and non-inferiority stopping rules and checked for type I and II errors. Interim analyses were done in the intention-to-treat population. This trial was registered with ClinicalTrials.gov, NCT02897076. Between Jan 2, 2017, and Oct 9, 2019, 3244 women were randomly assigned to the half-dose (n=1620 [49·9%]) or the full-dose group (n=1624 [50·1%]); 48 women withdrew consent, 30 fetuses were stillborn, 16 neonates were lost to follow-up, and 9 neonates died before evaluation, so that 3141 neonates remained for analysis. In the intention-to-treat analysis, the primary outcome occurred in 313 (20·0%) of 1567 neonates in the half-dose group and 276 (17·5%) of 1574 neonates in the full-dose group (risk difference 2·4%, 95% CI -0·3 to 5·2); thus non-inferiority was not shown. The per-protocol analysis also did not show non-inferiority (risk difference 2·2%, 95% CI -0·6 to 5·1). No between-group differences appeared in the rates of neonatal death, grade 3-4 intraventricular haemorrhage, stage ≥2 necrotising enterocolitis, severe retinopathy of prematurity, or bronchopulmonary dysplasia. Because non-inferiority of the half-dose compared with the full-dose regimen was not shown, our results do not support practice changes towards antenatal betamethasone dose reduction. French Ministry of Health.


1,25-dihydroxyvitamin D3 induction of nerve growth factor in L929 mouse fibroblasts: effect of vitamin D receptor regulation and potency of vitamin D3 analogs.
Decreased nerve growth factor (NGF) synthesis in the hippocampus and reduced nerve growth factor receptor immunoreactivity in CH1-4 basal forebrain areas have been implicated in neurodegeneration. Vitamin D receptors (VDR) have been located in brain areas affected by neurodegenerative diseases. 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3], the active form of vitamin D, has been shown to induce NGF in L929 mouse fibroblasts and rat hippocampus. In the present study we analyzed the VDR in L929 cells, which we used as a model system. We studied the regulation of VDR abundance and the ability of 1,25-(OH)2D3 to induce NGF synthesis. Scatchard analysis of [3H]1,25-(OH)2D3 binding showed the VDR concentration to be 173 fmol/mg protein and the affinity to be 0.12 nM. VDR was localized to nuclei of L929 cells by immunocytochemistry. Treatment of cells with forskolin (FSK; 50 microM), which activates the cAMP-protein kinase A pathway, resulted in an 8- to 10-fold up-regulation of VDR by 6 h, and VDR remained elevated at 24 h, as we have reported for other cells. NGF secretion was measured in serum-free conditioned medium using a double sided enzyme-linked immunosorbent assay. 1,25-(OH)2D3 treatment (0.1 pM to 10 nM) for 24 h increased the NGF concentration 2- to 3-fold, an effect that plateaued at 1 nM 1,25-(OH)2D3. VDR up-regulation by FSK pretreatment augmented the NGF response to 1,25-(OH)2D3 2-fold compared to that in vehicle-pretreated cells for a total 6-fold increase compared to basal NGF levels. The vitamin D analogs EB-1089 and 22-oxacalcitriol, which have been found to be less calcemic than 1,25-(OH)2D3, also induced NGF synthesis. The effects of these analogs were further enhanced by prior up-regulation of VDR with FSK. In conclusion, we have characterized the VDR in L929 cells and shown that 1,25-(OH)2D3 and its less calcemic analogs induce NGF. Furthermore, up-regulation of VDR abundance enhanced NGF induction. These effects of 1,25-(OH)2D3 and its analogs via VDR to regulate NGF synthesis may have significance for the eventual treatment of neurodegenerative diseases that are caused by decreased NGF production.


Simultaneous methanogenesis and denitrification of pretreated effluents from a fish canning industry.
A lab-scale hybrid upflow sludge bed-filter (USBF) reactor was employed to carry out methanogenesis and denitrification of the effluent from an anaerobic industrial reactor (EAIR) in a fish canning industry. The reactor was initially inoculated with methanogenic sludge and there were two different operational steps. During the first step (Step I: days 1-61), the methanogenic process was carried out at organic loading rates (OLR) of 1.0-1.25 g COD l-1 d-1 reaching COD removal percentages of 80%. During the second step (Step II: days 62-109) nitrate was added as KNO3 to the industrial effluent and the OLR was varied between 1.0 and 1.25 g COD l-1 d-1. Two different nitrogen loads of 0.10 and 0.22 g NO3(-)-N l-1 d-1 were applied and these led to nitrogen removal percentages of around 100% in both cases and COD removal percentages of around 80%. Carbon to nitrogen ratio (C:N) in the influent was maintained at 2.0 and eventually it was increased to 3.0, by means of glucose addition, to control the denitrification process. From these results it is possible to establish that wastewater produced in a fish canning industry can be used as a carbon source for denitrification and that denitrifying microorganisms were present in the initially methanogenic sludge. Biomass productions of 0.23 and 0.61 g VSS:g TOC fed for Steps I and II, respectively, were calculated from carbon global balances, showing an increase in biomass growth due to denitrification.


Monolayer contact doping of silicon surfaces and nanowires using organophosphorus compounds.
Monolayer Contact Doping (MLCD) is a simple method for doping of surfaces and nanostructures(1). MLCD results in the formation of highly controlled, ultra shallow and sharp doping profiles at the nanometer scale. In MLCD process the dopant source is a monolayer containing dopant atoms. In this article a detailed procedure for surface doping of silicon substrate as well as silicon nanowires is demonstrated. Phosphorus dopant source was formed using tetraethyl methylenediphosphonate monolayer on a silicon substrate. This monolayer containing substrate was brought to contact with a pristine intrinsic silicon target substrate and annealed while in contact. Sheet resistance of the target substrate was measured using 4 point probe. Intrinsic silicon nanowires were synthesized by chemical vapor deposition (CVD) process using a vapor-liquid-solid (VLS) mechanism; gold nanoparticles were used as catalyst for nanowire growth. The nanowires were suspended in ethanol by mild sonication. This suspension was used to dropcast the nanowires on silicon substrate with a silicon nitride dielectric top layer. These nanowires were doped with phosphorus in similar manner as used for the intrinsic silicon wafer. Standard photolithography process was used to fabricate metal electrodes for the formation of nanowire based field effect transistor (NW-FET). The electrical properties of a representative nanowire device were measured by a semiconductor device analyzer and a probe station.


Frictional Magneto-Coulomb Drag in Graphene Double-Layer Heterostructures.
Coulomb interaction between two closely spaced parallel layers of conductors can generate the frictional drag effect by interlayer Coulomb scattering. Employing graphene double layers separated by few-layer hexagonal boron nitride, we investigate density tunable magneto- and Hall drag under strong magnetic fields. The observed large magnetodrag and Hall-drag signals can be related with Laudau level filling status of the drive and drag layers. We find that the sign and magnitude of the drag resistivity tensor can be quantitatively correlated to the variation of magnetoresistivity tensors in the drive and drag layers, confirming a theoretical formula for magnetodrag in the quantum Hall regime. The observed weak temperature dependence and ∼B^{2} dependence of the magnetodrag are qualitatively explained by Coulomb scattering phase-space argument.


Perceived efficacy of COVID-19 restrictions, reactions and their impact on mental health during the early phase of the outbreak in six countries.
The COVID-19 pandemic forced millions of people to drastically change their social life habits as governments employed harsh restrictions to reduce the spread of the virus. Although beneficial to physical health, the perception of physical distancing and related restrictions could impact mental health. In a pre-registered online survey, we assessed how effective a range of restrictions were perceived, how severely they affected daily life, general distress and paranoia during the early phase of the outbreak in Brazil, Colombia, Germany, Israel, Norway and USA. Most of our over 2000 respondents rated the restrictions as effective. School closings were perceived as having the strongest effect on daily life. Participants who believed their country reacted too mildly perceived the risk of contracting SARS-CoV-2 to be higher, were more worried and expressed reduced beliefs in the ability to control the outbreak. Relatedly, dissatisfaction with governmental reactions corresponded with increased distress levels. Together, we found that satisfaction with one's governmental reactions and fear appraisal play an important role in assessing the efficacy of restrictions during the pandemic and their related psychological outcomes. These findings inform policy-makers on the psychological factors that strengthen resilience and foster the well-being of citizens in times of global crisis.


Toxicity of used orthodontic archwires assessed by three-dimensional cell culture.
The aim of the present study was to determine whether used orthodontic wires made of different materials cause toxicity and loss of viability on three-dimensional (3D) cell cultures. Three types of orthodontic wires, stainless steel, Nitinol, and TMA (n = 9) which had been used clinically in fixed appliances for a period of 1 month, were retrieved at random from five patients. Both upper and lower archwires were collected and subjected to two different protocols: to assess toxicity, two pieces of each wire were placed on 3D cell cultures (reconstituted human epithelium); to investigate the possibility of cell damage, the 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay was used and haematoxylin and eosin staining was performed to evaluate morphological changes. Copper wire served as the control to determine the morphology of severe toxicity, and native cell cultures and silk were used as the negative controls. Morphological evaluation of the native cell cultures revealed no toxic reactions. The ranking, from mild to severe toxicity was as follows: stainless steel < Nitinol = TMA. There were no significant differences between TMA and Nitinol. The MTT assay revealed the following mean percentage values for viability: native cell line (negative control), 100; stainless steel, 102.25; TMA, 87.4; Nitinol, 85.3; and copper wire (positive control) 57.2. Histological evaluation of the 3D cell cultures showed no severe toxicity or loss of viability for any of the wires. However, relative comparison between the different wires revealed that stainless steel induced less toxicity/loss of viability compared with TMA and Nitinol wire.


A Heterochromatin-Specific RNA Export Pathway Facilitates piRNA Production.
PIWI-interacting RNAs (piRNAs) guide transposon silencing in animals. The 22-30 nt piRNAs are processed in the cytoplasm from long non-coding RNAs that often lack RNA processing hallmarks of export-competent transcripts. By studying how these transcripts achieve nuclear export, we uncover an RNA export pathway specific for piRNA precursors in the Drosophila germline. This pathway requires Nxf3-Nxt1, a variant of the hetero-dimeric mRNA export receptor Nxf1-Nxt1. Nxf3 interacts with UAP56, a nuclear RNA helicase essential for mRNA export, and CG13741/Bootlegger, which recruits Nxf3-Nxt1 and UAP56 to heterochromatic piRNA source loci. Upon RNA cargo binding, Nxf3 achieves nuclear export via the exportin Crm1 and accumulates together with Bootlegger in peri-nuclear nuage, suggesting that after export, Nxf3-Bootlegger delivers precursor transcripts to the piRNA processing sites. These findings indicate that the piRNA pathway bypasses nuclear RNA surveillance systems to export unprocessed transcripts to the cytoplasm, a strategy also exploited by retroviruses.


A cytogenetic study in 120 Turkish children with intellectual disability and characteristics of fragile X syndrome.
We review the evidence for the frequency of the fragile X syndrome (FXS), other X-linked abnormalities, and other chromosomal disabilities of Turkish pediatric psychiatry outpatients with intellectual disability. Reported clinical features and genetic findings were used in cytogenetic screenings to estimate the prevalence of the fragile X (fra X) and other chromosomal aberrations in 120 patients with mental retardation, language disorders, attention deficit hyperactivity, or developmental delay, in comparison with 30 healthy children. Data on the clinical, intellectual and behavioral findings in 14 fra X positive children (11.7%) is presented. Ten of the 120 patients (8.3%) had enlargement of the heterochromatin region of chromosome 9. Other chromosomal aberrations and autosomal fragile sites (FS) were also observed. There was a statistically significant difference in the autosomal and X-linked FS between the study and control groups (p < 0.05). The tests for the fra X chromosome are likely to be of diagnostic benefit in young children with autism or developmental delay, particularly in speech, and who have large and prominent ears.


Impact of bariatric surgery on oral health conditions: 6-months cohort study.
We evaluate oral health conditions before and after bariatric surgery. The sample was composed of 59 patients who had undergone Roux-en-Y gastric bypass (RYGB). Salivary flow, periodontal pocket depth and dental wear were evaluated before and after 6 months of surgery. Body mass index (BMI), C-reactive protein (CRP) and glucose levels were obtained from the patient's medical files. A t-test was used for dependent samples. The mean BMI decreased from 49.31 ± 8.76 to 35.52 ± 8.12 kg/m(2) in 6 months after surgery (P < 0.000). Before surgery, 67% of patients had high levels of CRP and 38% higher blood glucose levels and after surgery there were significant reductions in these levels (P < 0.001). Salivary flow ranged from 0.84 to 0.95 ml/min. There was increased prevalence of periodontal pockets (P = 0.022) and mean pocket depth increased to about 0.5 mm (P < 0.001). The percentage of surfaces with dental wear in dentine was significantly higher after bariatric surgery (P = 0.002), while dental wear in enamel decreased (P = 0.019). Bariatric surgery may improve systemic conditions. However, it had a negative impact on oral health conditions because of an increase in periodontal disease and dental wear.


Novel dual-color immunohistochemical methods for detecting ERG-PTEN and ERG-SPINK1 status in prostate carcinoma.
Identification of new molecular markers has led to the molecular classification of prostate cancer based on driving genetic lesions. The translation of these discoveries for clinical use necessitates the development of simple, reliable and rapid detection systems to screen patients for specific molecular aberrations. We developed two dual-color immunohistochemistry-based assays for the simultaneous assessment of ERG-PTEN and ERG-SPINK1 in prostate cancer. A total of 232 cases from 184 localized and 48 metastatic prostate cancers were evaluated for ERG-PTEN and 284 cases from 228 localized and 56 metastatic prostate cancers were evaluated for ERG-SPINK1. Of the 232 cases evaluated for ERG-PTEN, 81 (35%) ERG-positive and 77 (33%) PTEN-deleted cases were identified. Of the 81 ERG-positive cases, PTEN loss was confirmed in 35 (15%) cases by fluorescence in situ hybridization (FISH). PTEN status was concordant in 203 cases (sensitivity 90% and specificity 87%; P<0.0001) by both immunohistochemisty and FISH; however, immunohistochemisty could not distinguish between heterozygous and homozygous deletion status of PTEN. Of the 284 cases evaluated for ERG-SPINK1, 111 (39%) cases were positive for ERG. In the remaining 173 ERG-negative cases, SPINK1 was positive in 26 (9%) cases. SPINK1 expression was found to be mutually exclusive with ERG expression; however, we identified two cases, of which one showed concomitant expression of ERG and SPINK1 in the same tumor foci, and in the second case ERG and SPINK1 were seen in two independent foci of the same tumor nodule. Unlike the homogenous ERG staining in cancer tissues, heterogeneous SPINK1 staining was observed in the majority of the cases. Further studies are required to understand the molecular heterogeneity of cases with concomitant ERG-SPINK1 expression. Automated dual ERG-PTEN and ERG-SPINK1 immunohistochemisty assays are simple, reliable and portable across study sites for the simultaneous assessment of these proteins in prostate cancer.


Beneficial effects of intraovarian injection of platelet-rich plasma in women with poor ovarian response.
Infertility can result from a diminished ovarian reserve, but a potential remedy exists in the form of platelet-rich plasma (PRP) administration. This treatment involves both biological factors and tissue trauma mechanisms, which stimulate folliculogenesis, making it a promising and effective strategy. We assessed the impact of direct PRP injections into the ovaries on the fertility outcomes of women classified as poor responders. A quasi-experimental study was conducted from April 2021 to December 2022, focusing on patients classified as POSEIDON grade 3 or 4. PRP injections were administered into both ovaries. After 3 months, data were collected on anti-Müllerian hormone (AMH) level, follicle-stimulating hormone (FSH) level, and the numbers of oocytes, mature oocytes, and good-quality embryos following ovarian stimulation. We then compared the data from before and after PRP injection. This study included 50 women, with a mean of 39 years (interquartile range [IQR], 35 to 43) and 4 years (IQR, 2 to 6) for age and infertility duration, respectively. FSH levels decreased after treatment, while AMH levels and the numbers of oocytes, metaphase II oocytes, and high-quality embryos increased. However, only the increase in high-quality embryos was significant. The pregnancy and spontaneous pregnancy rates were 20% and 14%, respectively. Notably, women with secondary infertility exhibited a significantly higher pregnancy rate than those with primary infertility. Ample evidence suggests that PRP can enhance ovarian function. However, further studies are needed to identify the appropriate candidates for this procedure, establish the optimal PRP preparation method, and standardize the procedure for its adjuvant use in assisted reproductive technology cycles.


Window of the World: Sino-African Encounters through Copies and Simulations.
Located in the manufacturing hub of Shenzhen is Window of the World: a Chinese theme park that features miniature copies of heritage sites from around the world. The individuals living within this constructed simulation are imported from diverse countries. They come to work as performers, animating the different cultural pavilions. As such, the transnationalism made possible by this park provides a window through which we can observe cultural interactivity, as well as the ways in which culture is constructed and re-presented. This article examines some of the processes of cultural encounters through copies of commodified cultural heritage. It also sheds light on the ways in which Kung Fu movies circulating in Africa have inspired the imaginations of young people, revealing cultural feedback loops that provide openings for new contact. Grounded in ethnographic research, the findings here are based on interviews with Kenyan and South African contract workers at this theme park. This article explores young people's pursuits of new opportunities of identity-formation and self-representation, as well as economic stability and forward mobility.


MICROTUBULE-ASSOCIATED PROTEIN65 is essential for maintenance of phragmoplast bipolarity and formation of the cell plate in Physcomitrella patens.
The phragmoplast, a plant-specific apparatus that mediates cytokinesis, mainly consists of microtubules (MTs) arranged in a bipolar fashion, such that their plus ends interdigitate at the equator. Membrane vesicles are thought to move along the MTs toward the equator and fuse to form the cell plate. Although several genes required for phragmoplast MT organization have been identified, the mechanisms that maintain the bipolarity of phragmoplasts remain poorly understood. Here, we show that engaging phragmoplast MTs in a bipolar fashion in protonemal cells of the moss Physcomitrella patens requires the conserved MT cross-linking protein MICROTUBULE-ASSOCIATED PROTEIN65 (MAP65). Simultaneous knockdown of the three MAP65s expressed in those cells severely compromised MT interdigitation at the phragmoplast equator after anaphase onset, resulting in the collapse of the phragmoplast in telophase. Cytokinetic vesicles initially localized to the anaphase midzone as normal but failed to further accumulate in the next several minutes, although the bipolarity of the MT array was preserved. Our data indicate that the presence of bipolar MT arrays is insufficient for vesicle accumulation at the equator and further suggest that MAP65-mediated MT interdigitation is a prerequisite for maintenance of bipolarity of the phragmoplast and accumulation and/or fusion of cell plate-destined vesicles at the equatorial plane.


Consensus document on European brain research.
Brain disease psychiatric and neurologic disease combined represents a considerable social and economic burden in Europe. Data collected by the World Health Organization (WHO) suggest that brain diseases are responsible for 35% of Europe's total disease burden. An analysis of all health economic studies of brain diseases in Europe, published by the European Brain Council (EBC) in June 2005, estimated the total cost of brain disease in Europe in 2004 to be Euro 386 billion. That burden is set to grow, mainly due to the fact that the European population is ageing. Investment in brain sciences does not match that burden now, let alone in the future. Brain research received only 8% of the life science budget in the European Commission's Fifth Framework Programme, which represents less than 0.01% of the annual cost of brain disorders for that period. Over the last decade, Europe has been losing ground to the USA and Japan in terms of both basic and clinical research. Many of Europe's young researchers are taking up posts in the USA and staying there. Big pharmaceutical companies are fleeing Europe for the USA, taking their drug development programmes with them. Research in the brain sciences now holds the promise of therapies that halt and even reverse neurodegeneration, of better diagnostic tools, neural prostheses for the paralysed and drugs for depression and anxiety that are tailored to the individual, thereby eliminating or reducing side effects. Our growing understanding of the normal brain could lead to better prevention of brain disease and to more effective teaching methods. The need for innovative treatments has never been greater, and Europe boasts clusters of excellent researchers in biotechnology who could collaborate with brain scientists and the pharmaceutical industry to realise this promise. But if Europe is to seize these opportunities and meet the challenge of brain disease, it needs to go forward on the basis of greater collaboration between countries, greater collaboration between industry, academia and patient organisations, and increased investment in the brain sciences. The EBC was formed in 2002 to bring together scientists, clinicians, the pharmaceutical industry, charities and patient organisations from all over Europe to campaign for these goals. It takes a novel, bottom-up approach to research policy, and in developing this consensus document, it aims to promote a greater and more focused effort in this area, to improve public understanding of the brain sciences and above all, to support brain research as a priority under the European Commission's Seventh Framework Programme (FP7, 2007-2013). The research programme outlined here was first conceived by the EBC board. An outline was sent to all member organisations and a number of individual experts for comments. Following that, a table of contents was developed. The 45 research themes were written by groups of experts from across Europe who represent a wide range of disciplines. Each one contains a proposal for future research on a specific brain-related theme which the EBC believes could form the basis of one or more integrated projects or strategic targeted research projects (STREP) funded under FP7. The EBC has deliberately focused on the major diseases and then described the basic research needed to understand and treat or perhaps even cure those diseases. The programme is therefore constructed "from man to molecule" and not the other way round, with equal importance attached to basic and clinical research. The EBC suggests that each of the proposed integrated projects or STREP should be awarded a budget in the order of Euro 10 to 15 million. In addition, brain research should be treated as an important element of many other parts of FP7, such as the European Research Council and research programmes on information technology and the causes of violence. Any research programme that concerns human behaviour should, by definition, take account of brain research. The EBC envisages that the priority for brain research it proposes at the European level will translate into higher priority for brain research at the national level, and this document may also serve as a starting point for the development of national consensus programmes. It seems likely that consensus conferences on brain research in Europe may further develop the themes and ideas discussed here. An EBC task force may also be established to further the consensus process. In general, increasing funding in the brain sciences would bring enormous economic returns by lightening the burden on healthcare systems and increasing the productivity of affected individuals-and might easily pay for itself. The human and social returns of such an investment are inestimable. And the time to act is now.


Spinal subdural haematoma: a rare complication of low-molecular-weight heparin therapy.
A 52-year-old man presented with chest pain, diarrhoea, rash, and arthritis. The use of low-molecular-weight heparin for suspected pulmonary embolism or angina led to a spinal subdural haematoma 3 days later. He was retrospectively confirmed to have Salmonella paratyphi infection. The clinical presentation and management of spinal subdural haematomas, and the incidence and manifestations of reactive arthritis related to Salmonella infections are briefly discussed.


Social skills and relapse history in outpatient schizophrenics.
Persons with schizophrenia commonly have impaired social functioning (Wallace 1984). Those with greater impairments, particularly as measured by premorbid social attainment, have a poorer clinical prognosis (Strauss and Carpenter 1972). Interventions designed to improve social competence, such as social skills training, have yielded generalizable and durable effects and may have reduced relapse rates (Wallace and Liberman 1985; Liberman et al. 1986; Hogarty et al. 1986). Thus, a valid measure of social skills should be a useful clinical and research tool. This research explores the validity of a new instrument for measuring such skills, the Assessment of Interpersonal Problem Solving Skills (AIPSS) (Donahoe et al., this issue). The AIPSS differs from more conventional social functioning measures because it 1) utilizes observations of role-playing, rather than self-report or third-party report; 2) provides a rating of the patient's current rather than past functioning; 3) involves videotaped simulated "real life" situations that pose challenges to the patient's ability to solve socially relevant problems; 4) permits assessment of patient's social perception, processing of social information for action planning, and verbal and nonverbal social responses. In this study, we examined the relationship between these parameters of schizophrenic patients' social functioning and their recent relapse history.


Optimizing combination chemotherapy by controlling drug ratios.
Cancer chemotherapy treatments typically employ drug combinations in which the dose of each agent is pushed to the brink of unacceptable toxicity; however, emerging evidence indicates that this approach may not be providing optimal efficacy due to the manner in which drugs interact. Specifically, whereas certain ratios of combined drugs can be synergistic, other ratios of the same agents may be antagonistic, implying that the most efficacious combinations may be those that utilize certain agents at reduced doses. Advances in nano-scale drug delivery vehicles now enable the translation of in vitro information on synergistic drug ratios into improved anticancer combination therapies in which the desired drug ratio can be controlled and maintained following administration in vivo, so that synergistic effects can be exploited. This "ratiometric" approach to combination chemotherapy opens new opportunities to enhance the effectiveness of existing and future treatment regimens across a spectrum of human diseases.


[Lysosomal hydrolase activity in the rat liver and serum depending upon the quality of dietary proteins].
Activity of lysosomal hydrolases cathepsins B, C, D and acid phosphatase was studied in liver tissue and blood serum of rats, ration of which contained 18% and 36% of the sunflower protein concentrate within one and three months. The control animals were maintained on rations with corresponding amount of casein. Activity of the proteinases studied tended to decrease in liver tissue of experimental animals, whereas activity of cathepsins C and D differed from the initial level after 3 months on the experimental diet. These alterations were dissimilar in blood serum: activity of cathepsin C was decreased within 1 month, while the enzyme activity as well as that of acid phosphatase was increased to the end of the experimental period (3 months). The data obtained suggest that lysosomal enzymes are sensitive to alterations in qualitative composition of the ration proteins; this property of the enzymes may serve as a criterion in medico-biological evaluation of new proteins in foodstuffs.


Quantitative Value of Aldosterone-Renin Ratio for Detection of Aldosterone-Producing Adenoma: The Aldosterone-Renin Ratio for Primary Aldosteronism (AQUARR) Study.
Current guidelines recommend use of the aldosterone-renin ratio (ARR) for the case detection of primary aldosteronism followed by confirmatory tests to exclude false-positive results from further diagnostic workup. We investigated the hypothesis that this could be unnecessary in patients with a high ARR value if the quantitative information carried by the ARR is taken into due consideration. We interrogated 2 large data sets of prospectively collected patients studied with the same predefined protocol, which included the captopril challenge test. We used an unambiguous diagnosis of aldosterone-producing adenoma as reference index. We also assessed whether the post-captopril ARR and plasma aldosterone concentration fall furnished a diagnostic gain over baseline ARR values. We found that the false-positive rate fell exponentially, and, conversely, the specificity increased with rising ARR values. At receiver operating characteristics curves and diagnostic odds ratio analysis, the high baseline ARR values implied very high positive likelihood ratio and diagnostic odds ratio values. The baseline and post-captopril ARR showed similar diagnostic accuracy (area under the receiver operating characteristics curve) in both the exploratory and validation cohorts, indicating lack of diagnostic gain with this confirmatory test (between-area under the curve difference, 0.005; 95% CI, -0.031 to 0.040; P=0.7 for comparison, and 0.05; 95% CI, -0.061 to 0.064; P=0.051 for comparison, respectively). These results indicate that the ARR conveys key quantitative information that, if properly used, can simplify the diagnostic workup, resulting in saving of money and resources. This can offer the chance of diagnosis and ensuing adrenalectomy to a larger number of hypertensive patients, ultimately resulting in better control of blood pressure.


Impact of synthetic noise signature and physiologic ECG signal on designing ML-based ECG noise detection framework.
Automatic signal analysis using artificial intelligence is getting popular in digital healthcare, such as ECG rhythm analysis, where ECG signals are collected from traditional ECG machines or wearable ECG sensors. However, the risk of using an automated system for ECG analysis when noise is present can lead to incorrect diagnosis or treatment decisions. A noise detector is crucial to minimise the risk of incorrect diagnosis. Machine learning (ML) models are used in ECG noise detection before clinical decision-making systems to mitigate false alarms. However, it is essential to prove the generalisation capability of the ML model in different situations. ML models performance is 50% lesser when the model is trained with synthetic and tested with physiologic ECG datasets compared to trained and tested with physiologic ECG datasets. This suggests that the ML model must be trained with physiologic ECG datasets rather than synthetic ones or add more various types of noise in synthetic ECG datasets that can mimic physiologic ECG.Clinical relevance- ML model trained with synthetic noisy ECG can increase the 50% misclassification rate in ECG noise detection compared to training with physiologic ECG datasets. The wrong classification of noise-free and noisy ECG will lead to misdiagnosis regarding the patient's condition, which could be a cause of death.


Are there any differences between features of proteins expressed in malignant and benign breast cancers?
The most common cancer among women is breast cancer and it has been blamed as the second leading cause of cancer death in women; so far many approaches have been used to analyze and detect benign and malignant forms of cancer and understanding the features involved in proteins expressed by various types of breast cancers is crucial. Herein features of proteins expressed in malignant, benign and both cancers were compared using different screening techniques, clustering methods, decision tree models and generalized rule induction (GRI) algorithms to look for patterns of similarity in two benign and malignant breast cancer groups. The findings showed that the N-terminal amino acid was Met and 57 out of 838 proteins' features ranked as important (p > 0.05). The depth of the trees induced by tree induction models varied from 5 (in the Quest model) to 2 (in the C5.0 model) branches. The best performance evaluation found when C&RT model applied and the worst evaluation found when CHAID model applied. No significant difference in the percentage of correctness, performance evaluation, and mean correctness in tree induction algorithms was found when feature selection applied on datasets, but the number of peer groups reduced significantly (p < 0.05) when feature selection model applied. The frequency of Ile-Ile was the most important protein attributes in all tree and rule induction models. The importance of sequence-based classification and the frequency of Ile-Ile in prediction of malignant and benign breast cancer have been discussed here.


Factor Structure of the Ways of Coping Questionnaire in Parkinson's Disease.
The Ways of Coping Questionnaire (WCQ) is used extensively in health research, but the measurement properties and suitability of the WCQ for people with Parkinson's disease (PD) have not been psychometrically assessed. If the WCQ does not align with its original 8-factor structure in a PD population, the use of the WCQ subscales may not be appropriate. The present study used confirmatory factor analysis (CFA), exploratory factor analysis (EFA), and multiple-group EFA to determine the ideal factor structure of the WCQ in a PD sample. The original 8 factors of the WCQ were not reproduced. EFA revealed a 6-factor structure, including Distancing, Faith, Avoidance, Seeking Social Support, Planful Problem Solving, and Confrontive coping. As motor symptom severity may impact coping, the stability of the 6-factor structure was examined across motor symptom severity (mild and moderate), remaining consistent. Higher levels of overall motor severity were associated with increased use of faith and avoidance style coping. These findings suggest that the 6-factor structure of the WCQ may be more appropriate for assessing coping styles in PD.


Current Management Practices in Ménière's Disease.
To evaluate current trends in managing Ménière's disease (MD) by both general otolaryngologists and otologists/neurotologists and discuss treatment modalities. Cross-sectional study. Survey of physicians. An electronic questionnaire was disseminated to all members of American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS). Eight hundred and sixty members replied for a response rate of 14.5% for generalists and 35% for neurotologists. Thirty-nine percent of respondents think that diet and lifestyle changes are effective in controlling symptoms in more than 50% of their MD patients. Overall, 72.8% of respondents used hydrochlorothiazide/triamterene (HCTZ/TAT) "often" or "always" with neurotologists using HCTZ/TAT more often than generalists (p < 0.001). Half of neurotologists used IT steroids "often" or "always," compared with only 10% of generalists (p < 0.001). Endolymphatic sac procedures are the most common surgeries and are used more often by neurotologists than by generalists (p < 0.001). The Medtronic Meniett device is used more by neurotologists (p < 0.001) but it is not commonly used overall (69.2% "never" use). Many options are available for the treatment of MD. Neurotologists tend to use a wider variety of medications in their treatment protocols than generalists. Neurotologists tend to perform surgical interventions more frequently than generalists. Our evidence shows significant heterogeneity for treatment of Ménière's disease among otolaryngologists. A guideline that outlines appropriate therapeutic options, dosing, and treatment escalation is warranted.


Tricyclic and related drugs for nocturnal enuresis in children.
Enuresis (bedwetting) is a socially disruptive and stressful condition which affects around 15-20% of five year olds, and up to 2% of young adults. To assess the effects of tricyclic and related drugs on nocturnal enuresis in children, and to compare them with other interventions. We searched the Cochrane Incontinence Group trials register (December 2002) and the reference lists of relevant articles including two previously published versions of this review. Date of the most recent searches: December 2002. All randomised and quasi-randomised trials of tricyclics or related drugs for nocturnal enuresis in children were included in the review. Comparison interventions included placebo, other drugs, alarms, behavioural methods or complementary/miscellaneous interventions. Trials focused solely on daytime wetting were excluded. Two reviewers independently assessed the quality of the eligible trials, and extracted data. Fifty four randomised trials met the inclusion criteria, involving 3379 children. The quality of many of the trials was poor. Most comparisons or outcomes were addressed only by single trials. Treatment with most tricyclic drugs (such as imipramine, amitriptyline, viloxazine, nortriptyline, clomipramine and desipramine) was associated with a reduction of about one wet night per week while on treatment (eg imipramine compared with placebo, weighted mean difference (WMD) -1.19, 95% CI -1.56 to -0.82). The exception was mianserin, where results from one small trial did not reach statistical significance. About a fifth of the children became dry while on treatment (relative risk for failure (RR) 0.77, 95% CI 0.72 to 0.83), but this effect was not sustained after treatment stopped (eg imipramine versus placebo, RR 0.98, 95% CI 0.95 to 1.03). There was not enough information to assess the relative performance of one tricyclic against another, except that imipramine was better than mianserin. The evidence comparing desmopressin with tricyclics was unreliable or conflicting, but in one small trial all the children failed or relapsed after stopping active treatment with either drug.The evidence comparing tricyclics with alarms was also unreliable or conflicting during treatment. In one small trial all the children failed or relapsed after tricyclics stopped, compared with about half after alarms. This result was compatible with the results in the Cochrane review of alarm treatment, which found that about half the children remained dry after alarm treatment was finished. There was a little evidence from single trials to suggest that imipramine might be better than a simple reward system with star charts during treatment; worse than a complex intervention involving education, counseling, waking and retention control training; better than a restricted diet; and worse than hypnosis. However, these results need to be confirmed by further research. Although tricyclics and desmopressin are effective in reducing the number of wet nights while taking the drugs, most children relapse after stopping active treatment. In contrast, only half the children relapse after alarm treatment. Parents should be warned of the potentially serious adverse effects of tricyclic overdose when choosing treatment. Further research is needed into comparisons between drug and behavioural or complementary treatments, and should include relapse rates after treatment is finished.


Cyclin-dependent kinase inhibitor seliciclib shows in vitro activity in diffuse large B-cell lymphomas.
Despite recent improvements in treatment, a significant fraction of patients with diffuse large B-cell lymphoma (DLBCL) still fail therapy. Therefore, new therapeutic modalities are needed to advance the cure rate. Seliciclib (CYC202, R-roscovitine) is a purine analog developed as an inhibitor of CDK2/cyclin E CDK7/cyclin H and CDK9/cyclin T. Seliciclib has been shown to be active in B-cell neoplasms, such as mantle cell lymphoma, chronic lymphocytic leukemia and in multiple myeloma in vitro. The aim of this study was to assess the in vitro activity of seliciclib in DLBCL. The anti-proliferative activity of seliciclib was tested in nine human DLBCL cell lines and six DLBCL primary cell cultures. The effects of seliciclib on the cell cycle and on apoptosis, as well as on transcription-related proteins were assessed. The cell viability of all DLBCL cell lines and primary cells was reduced by seliciclib treatment. The IC50 for the cell lines ranged from 13 - 36 microm. The effect of seliciclib was independent of the genetic aberrations characterizing the cell lines. After seliciclib exposure cells accumulated in G2/M or in G1 phase, with most of the cells showing signs of apoptosis. Despite the clear cytotoxic effect and induction of apoptosis, this study could not identify a unique mechanism of action. The in vitro data suggest that seliciclib is an active agent in DLBCL. Its efficacy is apparently independent of the underlying chromosomal translocations characteristic of DLBCL. The drug might represent a new therapeutic agent in this lymphoma sub-type.


TNF-alpha increases ceramide without inducing apoptosis in alveolar type II epithelial cells.
Ceramide is a bioactive lipid mediator that has been observed to induce apoptosis in vitro. The purpose of this study was to determine whether endogenous ceramide, generated in response to in vivo administration of tumor necrosis factor-alpha (TNF-alpha), increases apoptosis in primary rat alveolar type II epithelial cells. Intratracheal instillation of TNF-alpha (5 microgram) produced a decrease in sphingomyelin and activation of a neutral sphingomyelinase. These changes were associated with a significant increase in lung ceramide content. TNF-alpha concomitantly activated the p42/44 extracellular signal-related kinases and induced nuclear factor-kappaB activation in the lung. Hypodiploid nuclei studies revealed that intratracheal TNF-alpha did not increase type II cell apoptosis compared with that in control cells after isolation. A novel observation from separate in vitro studies demonstrated that type II cells undergo a gradual increase in apoptosis after time in culture, a process that was accelerated by exposure of cells to ultraviolet light. However, culture of cells with a cell-permeable ceramide, TNF-alpha, or a related ligand, anti-CD95, did not increase apoptosis above the control level. The results suggest that ceramide resulting from TNF-alpha activation of sphingomyelin hydrolysis might activate the mitogen-activated protein kinase and nuclear factor-kappaB pathways without increasing programmed cell death in type II cells.


Test-retest reproducibility of dopamine D2/3 receptor binding in human brain measured by PET with [11C]MNPA and [11C]raclopride.
Dopamine D receptors (DRs) have two affinity states for endogenous dopamine, referred to as high-affinity state (D ), which has a high affinity for endogenous dopamine, and low-affinity state (D ). The density of D can be measured with (R)-2-CHO-N-n-propylnorapomorphine ([C]MNPA), while total density of D and D (DRs) can be measured with [C]raclopride using positron emission tomography (PET). Thus, the ratio of the binding potential (BP) of [C]MNPA to that of [C]raclopride ([C]MNPA/[C]raclopride) may reflect the proportion of the density of D to that of DRs. In the caudate and putamen, [C]MNPA/[C]raclopride reflects the proportion of the density of D to that of DRs. To evaluate the reliability of the PET paradigm with [C]MNPA and [C]raclopride, we investigated the test-retest reproducibility of non-displaceable BP (BP ) measured with [C]MNPA and of [C]MNPA/[C]raclopride in healthy humans. Eleven healthy male volunteers underwent two sets of PET studies on separate days that each included [C]MNPA and [C]raclopride scans. BP values in the caudate and putamen were calculated. Test-retest reproducibility of BP of [C]MNPA and [C]MNPA/[C]raclopride was assessed by intra-subject variability (absolute variability) and test-retest reliability (intraclass correlation coefficient: ICC). The absolute variability of [C]MNPA BP was 5.30 ± 3.96 % and 12.3 ± 7.95 % and the ICC values of [C]MNPA BP were 0.72 and 0.82 in the caudate and putamen, respectively. The absolute variability of [C]MNPA/[C]raclopride was 6.11 ± 3.68 % and 11.60 ± 5.70 % and the ICC values of [C]MNPA/[C]raclopride were 0.79 and 0.80 in the caudate and putamen, respectively. In the present preliminary study, the test-retest reproducibility of BP of [C]MNPA and of [C]MNPA/[C]raclopride was reliable in the caudate and putamen.


Multicomponent reaction to construct spirocyclic oxindoles with a Michael (triple Michael)/cyclization cascade sequence as the key step.
Multicomponent cycloadditions with readily available isocyanides, allenoates, and isatylidene malononitriles are disclosed. This reaction, which does not require the aid of any catalyst, allows the efficient syntheses of spirocyclic oxindoles with excellent regioselectivity. Reactions with ethyl 2,3-butadienoate and various structurally diverse α- and γ-substituted allenoates are also fully explored. Remarkably, we have shown that the usual three-component process can be further developed into an unprecedented four-component cycloaddition in the presence of water, which provides a new strategy to access highly unusual tricyclic oxindoles. From a synthetic point of view, this protocol is very interesting considering the high level of complexity reached in one step. The mechanism is thought to proceed by a triple Michael/cyclization process by using allenoate as a three carbon atom component (3 C). Furthermore, multicomponent reaction with γ-substituted allenoate also results in a very interesting conversion. In such cases, the unusual cleavage of the "C=C" double bond of isatylidene malononitrile and one of the "C=C" double bonds of allenoate is always observed.


Absolute Quantification of Amyloid Propagons by Digital Microfluidics.
The self-replicating properties of proteins into amyloid fibrils is a common phenomenon and underlies a variety of neurodegenerative diseases. Because propagation-active fibrils are chemically indistinguishable from innocuous aggregates and monomeric precursors, their detection requires measurements of their replicative capacity. Here we present a digital amyloid quantitative assay (d-AQuA) with insulin as model protein for the absolute quantification of single replicative units, propagons. D-AQuA is a microfluidics-based technology that performs miniaturized simultaneous propagon-induced amplification chain reactions within hundreds to thousands of picoliter-sized droplets. At limiting dilutions, the d-AQuA reactions follow a stochastic regime indicative of the detection of single propagons. D-AQuA thus enables absolute quantification of single propagons present in a given sample at very low concentrations. The number of propagons quantified by d-AQuA was similar to that of fibrillar insulin aggregates detected by atomic-force microscopy and to an equivalent microplate-based assay, providing independent evidence for the identity of insulin propagons with a subset of morphologically defined protein aggregates. The sensitivity, precision, and accuracy of d-AQuA enable it to be suitable for multiple biotechnological and medical applications.


Immunological, genetic, and microbiological study of family members manifesting early-onset periodontitis.
From the standpoint of host-parasite interactions, family studies help us understand the host defensive factors and the molecular mechanisms involved in the periodontal immune response. In this study, we report the immunological profile of host-defensive functions, human leukocyte antigen (HLA) phenotypes, and the microflora of a mother (rapidly progressive periodontitis), an older son (periodontally healthy), a younger son (localized juvenile periodontitis), and a daughter (localized juvenile periodontitis). We examined the peripheral neutrophil functions, phenotypic and functional analysis of peripheral lymphocytes, serum immunoglobulin G (IgG) antibody titers against periodontopathic bacteria, serological type of HLA class II antigens, and bacterial flora in all periodontal pockets. The results showed that Actinobacillus actinomycetemcomitans was dominant in the pockets of all subjects. The mother and two sons showed a depressed neutrophil chemotaxis to N-formyl-methionyl-leucyl-phenylalanine. All subjects except the older son exhibited low T4/T8 ratios. The mother and daughter had raised levels of IgG titers to Porphyromonas gingivalis. All subjects had HLA phenotypes of DRw52 and DQ1 in common. We found that the family members had similar disorders in certain defensive functions. This family has been a model for our understanding of the host defensive factors in the development of early-onset periodontitis.


Cytological features of the Warthin-like variant of salivary mucoepidermoid carcinoma.
Warthin-like mucoepidermoid carcinoma is a recently proposed variant of musoepidermoid carcinoma. Histologically, it is characterized by its close resemblance to Warthin tumor, including dense lymphocytic infiltration, flattened intermediate epithelium resembling squamous metaplasia, and cystic change. Given its histologic similarity to Warthin tumor, confirmatory testing for MAML2 rearrangement is often required for this diagnosis. Here we present the first cytologic reports of two 53-year-old female patients with parotid masses. In both cases, the fine needle aspirations showed fragments of bland epithelium with a squamous appearance, mucinous cyst content, and focal lymphocytic background. Neither frank keratinization nor mucinous cells were identified in the smears. Fluorescence in situ hybridization (FISH) study confirmed MAML2 rearrangement on the resection specimens in both. Other cytologic differential diagnoses, including Warthin tumor with metaplasia, lymphadenoma, and lymphoepithelial cyst, were briefly discussed.


Role of arginine residues of D-aminoacylase from Alcaligenes xylosoxydans subsp. xylosoxydans A-6.
To investigate the role of arginine in the folding of d-aminoacylase, seven arginine residues, R26, R152, R296, R302, R354, R377, and R391, among twelve arginine residues highly conserved in d-aminoacylase, N-acyl-d-aspartate amidohydrolase (d-AAase), and N-acyl-d-glutamate amidohydrolase (d-AGase) from Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (Alcaligenes A-6) were substituted with lysine by site-directed mutagenesis. The mutants, R26K, R152K, R296K, and R302K were identified as mutations that increase partitioning of the enzyme into inclusion bodies. No mutants with substitutions within the carboxyterminal segment were found to increase partitioning into inclusion bodies (R354K, R377K, and R392K). These results suggest that arginine residues that position between the N-terminus and central region can play an important role in facilitating folding or stabilizing the structure of d-aminoacylase. By anaerobic cultivation, the production level of R302K in the soluble fraction was improved. Coexpression of the DnaK-DnaJ-GrpE chaperone assisted the folding of R302K, and reduced the effect of the aeration conditions on the solubility of R302K. We hypothesized that R302K requires a larger amount of chaperones for efficient folding than the wild type enzyme.


Cortico-NRM influences on trigeminal neuronal sensation.
We tested the idea that migraine triggers cause cortical activation, which disinhibits craniovascular sensation through the nucleus raphe magnus (NRM) and thus produces the headache of migraine. Stimulation of the dura mater and facial skin activated neurons in the NRM and the trigeminal nucleus. Stimulation of the NRM caused suppression of responses of trigeminal neurons to electrical and mechanical stimulation of the dura mater, but not of the skin. This suppression was antagonized by the iontophoretic application of the 5-HT(1B/1D) receptor antagonist GR127935 to trigeminal neurons. Migraine trigger factors were simulated by cortical spreading depression (CSD) and light flash. Activity of neurons in the NRM was inhibited by these stimuli. Multiple waves of CSD antagonized the inhibitory effect of NRM stimulation on responses of trigeminal neurons to dural mechanical stimulation but not to skin mechanical stimulation. The cortico-NRM-trigeminal neuraxis might provide a target for a more universally effective migraine prophylactic treatment.


Self-reported dry mouth in 50- to 80-year-old Swedes: Longitudinal and cross-sectional population studies.
Xerostomia is a common condition among elderly. The objectives were to examine prevalence, persistence, progression, yearly incidence of xerostomia, associated background factors and its influence on oral impacts on daily performances (OIDP) in 50- to 80-year-old people. In 1992, a questionnaire was sent to all 50-year-old (n = 8888) and in 2007 to all 75-year-old persons (n = 5195) living in two Swedish counties. In 2012, the same questionnaire was sent to both age cohorts, now 70- and 80-year-old. Response rate was for the 50-, 70- 75- and 80-year-old groups 71.4%, 72.2%, 71.9% and 66.4%, respectively. In the 50- to 70-year-old sample, 40.3% of the participants answered all five examinations and in the 75-80 group 49.5% (intact samples). In all age groups, xerostomia was significantly more prevalent in women than in men. At age 80, "often mouth dryness at night" was reported by 24.3% and 16.2% of women and men, respectively. Prevalence increased with age and was more frequent at night-time. Persistence of xerostomia was reported by 61.4%-77.5%, progression by 11.5%-33.0% and remission by 5.7%-11.3%. Average yearly incidence was 0.99%-3.28%. Xerostomia was more prevalent in those who reported a negative impact on OIDP. Highest odd ratios for xerostomia were burning mouth (OR 12.0), not feeling healthy (OR 5.1) and medicine usage (OR 3.9). Xerostomia is common in older age, persistence is high and progression common. The comorbidity between xerostomia, oral health problems and impaired general health needs to be taken into consideration when providing dental care to elderly patients.


A model for the biogenesis of turnip mosaic virus replication factories.
Nicotiana benthamiana plants were agroinfiltrated with an infectious clone of the Turnip mosaic virus (TuMV) that was engineered to tag replication vesicles with either GFP or mCherry fluorescent proteins. Punctuate vesicle structures were observed in the cytoplasm of infected cells corresponding to viral replication factories. The vesicles were highly motile and co-aligned with the microfilaments. Utilization of latrunculin B, an inhibitor of microfilament polymerization, reduced accumulation of the virus, suggesting that microfilaments are necessary during infection. To investigate biogenesis of the vesicles, leaves were infected simultaneously with two recombinant TuMV infectious clones, one that labeled vesicles in red and one that labeled them in green. We observed cell with green only and red only vesicles indicating a single viral genome origin. In some cases, vesicles exhibited sectors of green, red and yellow fluorescence were also observed, demonstrating that fusion among individual vesicles is possible. Based on those results we propose a model for the biogenesis of viral factory, where viral translation and replication are tightly coupled within virus-induced vesicles.


Maintaining a hip registry for 25 years. Mayo Clinic experience.
A computerized database was established for all total joint replacements done at the authors' institution. To date the registry contains information on more than 56,000 arthroplasties of which more than 30,000 involve the hip. The registry was designed to determine the effectiveness of total hip arthroplasty as a function of implant design, surgical technique, and patient selection. Furthermore, by maintaining and updating the patient record, data regarding success or anticipated failure could be communicated to the patient. Finally, this resource would provide reliable information that could be communicated to the orthopedic community. Patients are routinely evaluated at 1, 2, and 5 years postoperatively and at 5-year intervals thereafter by examination or letter or telephone questionnaire. Followup of patients at each interval is approximately 95%. Patients are more likely to respond by questionnaire (rather than be seen in person) if they are older, if a longer time has elapsed since surgery, or if they live a long distance from the clinic. Data are collected by five full time employees including computer and statistical support specifically assigned to the project. The annual joint registry budget is in excess of $400,000. Unfortunately, the future of this endeavor is challenged by the needs to: (1) show cost effectiveness of the activity; (2) update and validate outcomes instruments used as input into the database; and (3) maintain satisfactory followup rates in a medical economic environment that often discourages patient return visits or local assessment.


Prostaglandin E(1) is able to increase migration of leukocytes through endothelial cell monolayers.
Leukocyte interactions with endothelial cells play an important role during inflammatory processes. Leukocytes pass a monolayer of endothelial cells (ECM) to migrate into the extravascular space. The aim of the current study was to investigate whether prostaglandin E(1) (PGE(1)) influences the process of leukocyte migration. In a prospective controlled study, the influence of prostaglandin E(1) (50-5000 ng/mL) on leukocyte migration through endothelial cell monolayers (n = 7) was investigated. Human umbilical endothelial cells (HUVEC) and/or leukocytes were preincubated with clinically relevant, higher, and lower concentrations of prostaglandin E(1) and the amount of leukocyte migration after 3 h was measured. HUVEC were cultured on microporous membrane filters until achievement of a monolayer for investigation of leukocyte migration. Polymorphonuclear leukocytes (PMNL) were isolated from healthy volunteers and PMNL migration was studied under the influence of PGE(1). In clinically relevant concentrations, PGE(1) was able to increase significantly leukocyte migration through endothelial cell monolayers (205 +/- 7.8%, P < 0.05 compared to control; when treating PMNL alone, migration rate was 120 +/- 9.2% compared to control, ns; only endothelial cell monolayers treated up to 145 +/- 10.2%, P < 0.05 compared to control) showing a dose-dependent effect. In this assay, both cell types (PMNL and ECM) could be treated simultaneously, simulating the clinical situation after an iv administration. In conclusion, PGE(1) is able to increase leukocyte migration through endothelial cell monolayers when both cell types are pretreated. The treatment of either leukocytes or endothelial cell monolayers in the cell coculture showed no significant increase. These findings support the theory that prostaglandins may play a major role during inflammation. Future clinical studies are warranted to confirm this hypothesis.


The Need for Education and Clinical Best Practice Guidelines in the Era of Direct-to-Consumer Genomic Testing.
Many people share the results of their direct-to-consumer personal genomic testing (DTC-PGT) within the primary care setting, seeking interpretation of and counsel about the results. However, most primary care physicians (PCPs) are not trained to interpret and communicate about DTC-PGT results. New guidelines must be developed to help PCPs maximize the potential of emerging DTC-PGT technologies.


Assessment of Food Intake, Obesity, and Health Risk among the Homeless in Rhode Island.
The objective of this study was to examine the relationship between the nutritional status, incidence of food insecurity, and health risk among the homeless population in Rhode Island. This correlational study utilized a convenience sample of 319 homeless adults from Rhode Island's largest service agency for the homeless. Information on use of services such as access to emergency foods, shelters, and the Supplemental Nutrition Assistance Program (SNAP) was requested. Food security was measured by the six-item subset of the USDA Food Security Core Module. Anthropometric measures included height, weight, and waist circumference. A 24-hr dietary recall was collected to determine the food intake for a subset of participants who agreed to supply this information (n = 197). Average dietary recall data indicated insufficient intake of vegetables, fruit, dairy, and meats/beans. It also indicated excessive intake of fats. Of the 313 participants, 29.4% were overweight and 39% were obese. Over 94% of the participants were food insecure, with 64% of this subset experiencing hunger. Fifty-five percent of the participants were currently receiving SNAP benefits. The majority of the sample was found to be food insecure with hunger.


Cerebral venous thrombosis: Associations between disease severity and cardiac markers.
Plasma cardiac troponin (cTn) elevation occurs in acute ischemic stroke and intracranial hemorrhage and can suggest a poor prognosis. Because acute cerebral venous thrombosis (CVT) might lead to venous stasis, which could result in cardiac stress, it is important to evaluate whether cTn elevation occurs in patients with CVT. Inpatients at Johns Hopkins Hospital from 2005 to 2015 meeting the following criteria were included: CVT (ICD-9 codes with radiologic confirmation) and available admission electrocardiogram (ECG) and cTn level. In regression models, presence of ECG abnormalities and cTn elevation (>0.06 ng/mL) were evaluated as dependent variables in separate models, with location and severity of CVT involvement as independent variables, adjusted for age, sex, and hypertension. Of 81 patients with CVST, 53 (66%) met the inclusion criteria. Participants were, on average, aged 42 years, white (71%), and female (66%). The left transverse sinus was most commonly thrombosed (47%), with 66% having >2 veins thrombosed. Twenty-two (41%) had cTn elevation. Odds of cTn elevation increased per each additional vein thrombosed (adjusted OR 2.79, 95% CI [1.08-7.23]). Of those with deep venous involvement, 37.5% had cTn elevation compared with 4.4% without deep clots (p = 0.02). Venous infarction (n = 15) was associated with a higher mean cTn (0.14 vs 0.02 ng/mL, p = 0.009) and was predictive of a higher cTn in adjusted models (β = 0.15, 95% CI [0.06-0.25]). In this single-center cohort study, markers of CVT severity were associated with increased odds of cTn elevation; further investigation is needed to elucidate causality and significance.


Derepression of HMGA2 gene expression in retinoblastoma is associated with cell proliferation.
To assess whether retinoblastoma formation is associated with the expression of high mobility group (HMG) A2 protein, a transcription factor that is highly expressed during embryogenesis and completely repressed in normal adult tissues, we performed Northern and Western blots and RT-PCR analyses, and immunohistochemistry to test for HMGA2 expression. We used established retinoblastoma cell lines in tumors grown in nude mice and clinical retinoblastoma specimens, and contrasted these tumors with normal embryonic and adult retina. Adenoviral-mediated antisense experiments were conducted on the retinoblastoma cell lines to suppress HMGA2 expression and determine if cell proliferation is HMGA2-dependent. We also transfected a retinoblastoma cell line to identify cis-regulatory elements and transcription initiation sites on the HMGA2 gene promoter. HMGA2 gene expression was silenced in terminally differentiated retina of 6-wk-old mice, but it was detected in retina of a 13.5-d postcoitum embryo. Reactivation of HMGA2 gene expression was observed in the retinoblastoma cell lines Y79, WERI-Rb1, and TOTL-1, in tumors derived from some of these cells propagated in nude mice, and in a high frequency of retinoblastomas excised from human patients. This suggests that expression of HMGA2 gene in retinoblastoma cells involves a derepression process. By using an antisense approach to block HMGA2 expression, we observed a decrease in the number of proliferating retinoblastoma cells. As a 1st step toward understanding HMGA2 gene reactivation in retinoblastoma, we mapped the 2 transcription initiation sites and associated positive regulatory elements within the WERI-Rb1 cells. Our discovery of derepression of HMGA2 gene expression in retinoblastoma provides the 1st evidence that this protein might contribute to neoplastic transformation of retina cells.


[Nutritive value of raw and processed cowpeas].
Three experiments were carried out to determine the effects of autoclave cooking, extrusion and DL-methionine supplementation on the protein and energy value of cowpea. The two processes decreased the available lysine of cowpea from 6.3 to 5.7%. The starch content decreased from 50.7% in raw cowpea to 44.9% and 42.1% in cooked extruded cowpea, respectively. Cooking eliminated the trypsin inhibitor content of cowpea. Damaged starch increased from 0.3% in the raw cowpea, to 22.5% in cooked cowpea and to 30.5% in extruded cowpea. In the first experiment it was determined that the protein efficiency ratio (PER) in rats was not affected by cowpea processing (means = 1.17) and that DL-methionine supplementation increased (P less than 0.01) PER to 2.08. In the second experiment, it was established that energy digestibility of the diets used in the first experiment averaged 88.0% with no differences due to processing or supplementation. Digestible energy content (DE) was lower (P less than 0.01) in diets supplemented with DL-methionine (3.58, 3.56, 3.93) than in those without supplement (3.67, 3.93, 3.75). On the other hand, diets with extruded cowpea produced higher (P less than 0.01) DE (3.84 kcal/g) than diets with cooked cowpea (3.75 kcal/g). Classical (EM) and nitrogen corrected (EMn) metabolizable energy determined in chicks showed that both were higher (P less than 0.01) in cooked (3.01; 2.56 kcal/g) and extruded (3.16; 2.76 kcal/g) than in raw cowpeas (1.86; 1.81 kcal/g).


[Non cognitive symptoms in dementias].
In the phenomenology of dementia, the cognitive symptoms surround most of the interests both for investigators as clinicians. However, the non cognitive symptoms are shown so often they should become a major one in the clinical evaluation of the dementia syndrome. Moreover, the presence of this symptoms means more clinical severity, increases the institutionalization risk and causes a larger emotional burden for demented carers. On this work, the authors argue about the possible physiopathogenic causes related to cognitive and non cognitive aspects of dementia.


Therapeutic targets in age-related macular disease.
Age-related macular disease (AMD) accounts for more than 50% of blind registration in Western society. Patients with AMD are classified as having early disease, in which visual function is well preserved, or late disease, in which central vision is lost. Until recently, there was no therapy available by which the course of the disorder could be modified. Now, the most common form of late-stage AMD - choroidal neovascularization - responds to treatment with anti-VEGF therapies; although visual loss is modified in a portion of these cases, no therapeutic approach exists that alters the evolution from early to late disease. However, as discussed in this Review, research over the last few years has demonstrated several features of AMD that are likely to be amenable to treatment. Potential targets for treatment are described, and possible therapeutic approaches are discussed.