Two-center collaborative evaluation of performance of the BD phoenix automated microbiology system for identification and antimicrobial susceptibility testing of gram-negative bacteria.
The performance of the BD Phoenix Automated Microbiology System (BD Diagnostic Systems, Sparks, MD) was assessed for identification (ID) and antimicrobial susceptibility testing (AST) of the majority of clinically encountered bacterial isolates in a European collaborative two-center trial. A total of 494 bacterial isolates including various species of the Enterobacteriaceae and 110 nonfermentative gram-negative bacteria were investigated: of these, 385 were single patient isolates, and 109 were challenge strains tested at one center. The performance of the Phoenix extended-spectrum beta-lactamase (ESBL) test was also evaluated for 203 strains of Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca included in the study. Forty-two antimicrobial drugs were tested, including members of the following drug classes: aminoglycosides, beta-lactam antibiotics, beta-lactam/beta-lactamase inhibitors, carbapenems, cephems, monobactams, folate antagonists, quinolones, and others. Phoenix system ID results were compared to those of the laboratories' routine ID systems (API 20E and API CHE, ATB ID32E, ID32GN, and VITEK 2 [bioMérieux, Marcy l'Etoile, France]); Phoenix AST results were compared to those of frozen standard broth microdilution (SBM) panels according to NCCLS (now CLSI) guidelines (NCCLS document M100-S9, approved standard M7-A4). Discrepant results were repeated in duplicate. Concordant IDs of 98.4 and 99.1% were observed for the Enterobacteriaceae and the nonfermentative group, respectively. For AST results, the overall essential agreement was 94.2%; the category agreement was 97.3%; and the very major error rate, major error rate, and minor error rate were 1.6, 0.6, and 1.9%, respectively. In terms of ESBL detection, Phoenix results were 98.5% concordant with those of the reference system, with 98.0% sensitivity and 98.7% specificity. In conclusion, the Phoenix ID results showed high agreement with results of the systems to which they were being compared: the AST performance was highly equivalent to that of the SBM reference method, and the system proved to be very accurate for the detection of ESBL producers.


Studies on defined immunofluorescence in clinical immunopathology. III. Past progress, some new methods, and prospects for the future.
Experience with chessboard titrations over the past 9 years has shown that essentially all indirect IF staining systems that employ anti-IgG conjugates can be defined with the aid of assays for F/P ratios and/or antibody concentrations. Some improvements in the radial diffusion assays for antibody protein have been effected by refinements in methodology. Studies with a form of the Dass system showed that over a 16-fold range of light intensities, relatively little change occurred in "titers" of IgG (comparably to titers of patient sera) or in PEP conjugates as seen in DASS chessborad titrations. An analysis of 233 PEP values included in 12 literature reports on indirect IF tests indicate that 88% fall within one doubling dilution of 1/16 unit/ml or about 12.5 mug labeled Ab/ml. The DASS reactions also are within this range. Factors responsible for variation beyond these limits in indirect IF tests were analyzed. The most important factor appears to be experimental errors. Other factors include low titers of primary antibodies, some aberrant conjugates, and a recent shift toward greater sensitivity of chessboard titrations. The latter appears to be due primarily to modes of reading and should be controlled by the constant use of reference sera. It now seems possibe that future IF studies may be defined with the aid of this DASS system. To achieve this goal, it will be necessary to not only elaborate microanalytic techniques of the type that have been considered at this Conference but also to generate macroanalytic methods that can be used in parallel with them as a basis for interpreting microscopic observations both in the DASS system and in IF studies of sera and tissues.


In Situ Loading of Cu Nanocrystals on CsCuCl3 for Selective Photoreduction of CO2 to CH4.
Rational design and facile synthesis of efficient environmentally friendly all-inorganic lead-free halide perovskite catalysts are of great significance in photocatalytic CO2 reduction. Aiming at photogenerated charge carrier separation and CO2 reaction dynamics, in this paper, a CsCuCl3 /Cu nanocrystals (NCs) heterojunction catalyst is designed and synthesized via a simple acid-etching solution process by using Cu2 O as the sacrificed template. Due to the disproportionation reaction of Cu2 O induced by concentrated hydrochloric acid, Cu NCs can be deposited onto the surface of CsCuCl3 microcrystals directly and tightly. As revealed by photoelectrochemical analysis, in situ Fourier transform infrared spectra, etc., the Cu NCs contribute a lot to extracting photoelectrons of CsCuCl3 to improve the charge separation efficiency, regulating the CO2 adsorption and activation, and also stabilizing the reaction intermediates. Therefore, CsCuCl3 /Cu heterojunction exhibits a total electron consumption rate of 58.77 µmol g-1 h-1 , which is 2.9-fold of that of single CsCuCl3 . Moreover, high CH4 selectivity of up to 92.7% is achieved, which is much higher than that of CsCuCl3 (50.4%) and most lead-free halide perovskite-based catalysts. This work provides an ingenious but simple strategy to rationally design cocatalysts in situ decorated perovskite catalysts for manipulating both the catalytic activity and the product selectivity.


Scleral fixation of a single-piece foldable acrylic IOL through a 1.80 mm corneal incision.
A new scleral fixation technique of a single-piece acrylic foldable intraocular lens (IOL) (enVista MX60, Bausch & Lomb, Inc.) through a 1.80 mm corneal incision, using the IOL eyelets as anchoring point, is described. It was a retrospective review of 26 cases. The preoperative mean corrected distance visual acuity was 0.51 ± 0.21 logarithm of the minimum angle of resolution (logMAR). It improved significantly to 0.25 ± 0.27 logMAR (P < .01), 0.18 ± 0.16 logMAR (P < .01), and 0.17 ± 0.16 logMAR (P < .01) (at 1 month, 3 months, and 6 months postoperatively, respectively, repeated measures analysis of variance, P < .0001). No astigmatism increase of more than 0.75 diopters was recorded at any time point. In all 26 patients, the IOL was well centered and stable for the entire monitoring period. No complications were observed during follow-up. Scleral fixation of the foldable IOL through a 1.80 mm corneal incision provided excellent IOL stability during the 6-month follow-up of this study and might be an effective and safe surgical technique.


Junctate, an inositol 1,4,5-triphosphate receptor associated protein, is present in rodent sperm and binds TRPC2 and TRPC5 but not TRPC1 channels.
The acrosome reaction, the first step of the fertilization, is induced by calcium influx through Canonical Transient Receptor Potential channels (TRPC). The molecular nature of TRPC involved is still a debated question. In mouse, TRPC2 plays the most important role and is responsible for the calcium plateau. However, TRPC1 and TRPC5 are also localized in the acrosomal crescent of the sperm head and may participate in calcium signaling, especially in TRPC2-deficient mice. Activation of TRPC channels is an unresolved question in germ and somatic cells as well. In particular, in sperm, little is known concerning the molecular events leading to TRPC2 activation. From the discovery of IP3R binding domains on TRPC2, it has been suggested that TRPC channel activation may be due to a conformational coupling between IP3R and TRPC channels. Moreover, recent data demonstrate that junctate, an IP3R associated protein, participates also in the gating of some TRPC. In this study, we demonstrate that junctate is expressed in sperm and co-localizes with the IP3R in the acrosomal crescent of the anterior head of rodent sperm. Consistent with its specific localization, we show by pull-down experiments that junctate interacts with TRPC2 and TRPC5 but not with TRPC1. We focused on the interaction between TRPC2 and junctate, and we show that the N-terminus of junctate interacts with the C-terminus of TRPC2, both in vitro and in a heterologous expression system. We show that junctate binds to TRPC2 independently of the calcium concentration and that the junctate binding site does not overlap with the common IP3R/calmodulin binding sites. TRPC2 gating is downstream phospholipase C activation, which is a key and necessary step during the acrosome reaction. TRPC2 may then be activated directly by diacylglycerol (DAG), as in neurons of the vomeronasal organ. In the present study, we investigated whether DAG could promote the acrosome reaction. We found that 100 microM OAG, a permeant DAG analogue, was unable to trigger the acrosome reaction. Altogether, these results provide a new hypothesis concerning sperm TRPC2 gating: TRPC2 activation may be due to modifications of its interaction with both junctate and IP3R, induced by depletion of calcium from the acrosomal vesicle.


Evaluation of parental preference for the treatment of asthmatic children aged 6 to 11 years with oral montelukast or inhaled cromolyn: a randomized, open-label, crossover study.
The objective of this study was to evaluate parental preference for the treatment of asthmatic children with oral montelukast sodium or inhaled cromolyn sodium. Additionally, we wanted to compare the two drugs in terms of patient preference for treatment, patient and parent satisfaction with treatment, frequency of inhaled albuterol use, adherence to treatment, and safety. This was a 12-week randomized, open-label, crossover study conducted in 42 primary care and asthma/allergy specialty centers in the United States. Three hundred thirty-three asthmatic patients, ages 6 to 11 years, who had a forced expiratory volume in 1 second (FEV1) of 60%-85% (inclusive) of predicted value with > or = 12% reversibility after administration of an inhaled beta-agonist and who used albuterol on at least 7 of the last 14 days of the run-in period. After a 2- to 3-week run-in period, patients were randomized either to 4 weeks of montelukast (5-mg chewable tablet once daily) followed by a 2-week washout period, then 4 weeks of cromolyn (two puffs 4 times daily from a metered-dose inhaler) or to the reverse sequence. More parents preferred montelukast (87%) than cromolyn (12%; p < 0.001). More patients preferred montelukast (82%) than cromolyn (17%; p < 0.001). Daily albuterol use (puffs/day) was reduced by 38% during montelukast therapy vs. 23% during cromolyn therapy. Seventy-eight percent of patients reported being highly adherent to montelukast therapy compared with 42% to cromolyn therapy (p < 0.001). Fewer patients receiving montelukast discontinued because of asthma exacerbation (1.0% vs. 5.0%, respectively), and fewer patients reported worsening asthma while receiving montelukast (3.5% vs. 7.5%, p = 0.036). Parents' and patients' preference, parents' and patients' satisfaction, and patients' adherence to therapy were all significantly better with oral montelukast compared with inhaled cromolyn. Beta-agonist use was decreased when taking montelukast, which was safe and well-tolerated.


Model of lactose repressor core based on alignment with sugar-binding proteins is concordant with genetic and chemical data.
Using primary sequence similarity to arabinose-binding protein, D-glucose/D-galactose-binding protein, and ribose-binding protein (Vyas, N. K., Vyas, M. N., and Quiocho, F. A. (1991) J. Biol. Chem. 266, 5226-5237; Mowbray, S. L., and Cole, L. B. (1992) J. Mol. Biol. 225, 155-175), the core domain (residues 62-323) of the bacterial regulatory protein lac repressor has been aligned to these sugar-binding proteins of known structure. Although the sequence identity is not striking, there is strong overall homology based on two separate matrix scoring systems (minimum base change per codon (MBC/C) and amino acid homology per residue (AAH/R)) (mean score: MBC/C < 1.25, AAH/R > 5.50; random sequences: MBC/C = 1.45, AAH/R = 4.46). Similarly, the predicted secondary structure of the repressor exhibits excellent agreement with the known secondary structures of the sugar-binding proteins. Using this primary sequence alignment, the tertiary structure of the core domain of the lac repressor has been modeled based on the known structures of the sugar-binding proteins as templates. While the structure deduced for the repressor is hypothetical, the model generated allows a comparison between the predicted tertiary arrangement and the wealth of genetic and chemical data elucidated for the repressor. Important residues involved in operator and sugar binding and in protein assembly have been identified using genetic methods, and placement of these residues in the model is consistent with their known function. This approach, therefore, provides a means to visualize the core domain of the lac repressor that allows interpretation of genetic and chemical data for specific residues and rational design of future experiments.


Curli production and genetic relationships among Escherichia coli from cases of bovine mastitis.
Curli are adhesive surface structures produced by some Escherichia coli and Salmonella strains that bind host proteins and activate inflammatory mediators. In this study, 61 E. coli isolates from 36 clinical cases of bovine mastitis were characterized using enterobacterial repetitive intergenic consensus-PCR and screened for their ability to produce curli. Effect of curli production on case recovery, based on a return to precase milk yield, was investigated for a subset of 43 isolates from 20 quarters of 19 cows. Thirty-five (57%) of 61 isolates were curli positive. Fifty-eight of the 61 isolates clustered into 2 clonal groups at 52% genetic similarity. Genetically diverse E. coli isolates were simultaneously cultured from individual cases. Twenty-three isolates from 13 cows were clustered in clonal group I, of which 5 cases (38%) were curli positive; 35 isolates from 22 cows were clustered in clonal group II, of which 15 cases (68%) were curli positive. No association was found between genetic similarity and phenotypic curli expression of isolates from cows with clinical E. coli mastitis cases. Phenotypic curli expression in isolates did not affect recovery of cows' milk yield to premastitis production levels.


Single absorbable polydioxanone pin fixation for distal chevron bunion osteotomies.
The distal chevron osteotomy is a well-established technique for correction of symptomatic mild to moderate metatarsus primus varus with hallux valgus deformity. Fixation of the osteotomy ranges from none to bone pegs, Kirschner wires, screws, or absorbable pins. We evaluated one surgeon's (J.K.D.) results of distal chevron osteotomy fixation with a single, nonpredrilled, 1.3-mm poly-p-dioxanone pin and analyzed any differences in patients with unilateral or bilateral symptomatic metatarsus primus varus with hallux valgus deformities. All osteotomies healed without evidence of infection, osteolysis, nonunion, or necrosis. Equal correction was achieved in unilateral and bilateral procedures. The technique is quick and easy, and adequate fixation is achieved.


Sleep, sleep disorders and motor vehicle crashes.
There is wide consensus that drowsiness and sleep during driving put the driver and others at high risk for motor vehicle crashes. A large percentage of both private automobile and commercial vehicle crashes and crash fatalities are associated with the sleepy driver each year. Sleepiness in persons without sleep disorders may occur from potentially preventable causes such as sleep deprivation, drinking alcohol or taking sedative medications. Sleepiness also occurs uncontrollably in persons with certain sleep disorders. It is important, therefore, for persons to have a diagnosis of a sleep disorder, objectively confirmed with an MSLT, to be considered for restriction of driving privileges. The MSLT serves as a marker for the sleep disorder and also as evidence of effective treatment. While sleepiness is widespread, this regulation addresses only the highest risk group for attention. Proposed guidelines for sleep related disorders and driving in Wisconsin are presented with this background in mind.


Using an Ultrasound Tissue Phantom Model for Hybrid Training of Deep Learning Models for Shrapnel Detection.
Tissue phantoms are important for medical research to reduce the use of animal or human tissue when testing or troubleshooting new devices or technology. Development of machine-learning detection tools that rely on large ultrasound imaging data sets can potentially be streamlined with high quality phantoms that closely mimic important features of biological tissue. Here, we demonstrate how an ultrasound-compliant tissue phantom comprised of multiple layers of gelatin to mimic bone, fat, and muscle tissue types can be used for machine-learning training. This tissue phantom has a heterogeneous composition to introduce tissue level complexity and subject variability in the tissue phantom. Various shrapnel types were inserted into the phantom for ultrasound imaging to supplement swine shrapnel image sets captured for applications such as deep learning algorithms. With a previously developed shrapnel detection algorithm, blind swine test image accuracy reached more than 95% accuracy when training was comprised of 75% tissue phantom images, with the rest being swine images. For comparison, a conventional MobileNetv2 deep learning model was trained with the same training image set and achieved over 90% accuracy in swine predictions. Overall, the tissue phantom demonstrated high performance for developing deep learning models for ultrasound image classification.


Trachea enhances neurite regeneration from adult rat nodose ganglia in vitro.
Trachea is intensely innervated with vagal afferent nerve fibers, and may play an important role in vagus nerve regeneration after axonal injury caused by trauma and surgical operation. We investigated the effects of tracheal tissue on neuronal cell survival and neurite regeneration in adult rat nodose ganglia (NG) in vitro. Co-culture with trachea significantly increased the average number of neurites regenerated from transected nerve terminals of NG explants, from 73.7 to 154.2 after 3 days, from 68 to 186.7 after 5 days, and from 31 to 101.5 after 7 days in culture. Dissociated NG neurons could continue to survive and extend neurites only in the co-existence with satellite cells in collagen gel. Co-cultured trachea improved the ratios of survival and neurite-bearing cells of NG neurons, from 56.7% and 11.1% to 72.3% and 37.6% after 4 days, and from 41.1% and 20.3% to 56.4% and 47.2% after 7 days in culture, respectively. These results imply that tracheal tissue secretes a factor, which could enhance neuronal cell survival and neurite regeneration in NG in the presence of satellite cells in vitro.


Kidney transplant in black recipients: are African Europeans different from African Americans?
Despite recent improvement, significant racial disparities in outcome still persist after renal transplantation among African American patients in the United States. This study evaluated the association of race and ethnicity with allograft outcomes in a French population of 952 Caucasian (Cauc) patients and 140 African European (AE) patients who underwent renal transplantation in our center between 1987 and 2003. Demographic characteristics were similar for the two cohorts other than cause of end-stage renal failure (more hypertension among AE and more polycystic kidney disease among Cauc) and cold ischemia time (significantly longer for AE). Immunosuppressive treatment was comparable between groups. There were no significant differences between AE and Cauc in the incidence of acute rejection (31% vs. 30%). At 5 years post-transplant, patient survival (93% vs. 92%), graft survival (83% in both groups) and graft function (creatinine clearance 48 mL/min vs. 45 mL/min) were also similar among the AE and Cauc patients. We demonstrate that ethnic origin does not affect outcome after renal transplantation in France. Therefore, differences observed in the United States cannot be only related to immunologic or pharmacologic factors. The results of renal transplantation in patients of African origin could be improved with universal immunosuppressive drug coverage.


Poly(vinyl alcohol)-Poly(ethylene glycol) Double-Network Hydrogel: A General Approach to Shape Memory and Self-Healing Functionalities.
A double-network polymer hydrogel composed of chemically cross-linked poly(ethylene glycol) (PEG) and physically cross-linked poly(vinyl alcohol) (PVA) was prepared. When the hydrogel (70 wt % of water) is subjected to freezing/thawing treatment under strain, the enhanced physical network as a result of crystallization of PVA chains can stabilize the hydrogel deformation after removal of the external force at room temperature. Subsequent disruption of the physical network of PVA by heating allows for the recovery of the initial shape of the hydrogel. Moreover, the double-network hydrogel exhibits self-healing capability stemming from the physical network of PVA by virtue of the extensive interchain hydrogen bonding between the hydroxyl side groups. This study thus demonstrates a general approach to imparting both the shape memory and self-healing properties to chemically cross-linked hydrogels that otherwise do not have such functionalities. Moreover, by making use of the fixed hydrogel elongation, the effect of anisotropy arising from chain orientation on the self-healing was also observed.


[Elucidation of the Molecular Basis of Abnormal Prion Protein (PrP) Formation in a Cell-Free System Using Baculovirus and Insect Cell-derived Recombinant PrP].
The molecular basis underlying the conversion of normal prion protein (PrPC) into abnormal prion protein (PrPSc) has not been fully elucidated. The protein-misfolding cyclic amplification (PMCA) technique, which can amplify PrPSc in vitro with the use of intermittent sonication, mimics the process of in vivo PrPSc replication. Accumulating evidence suggests that co-factors other than PrP may play a crucial role in the faithful replication of PrPSc. In conventional PMCA, brain homogenates (BHs) from normal animals are used as the PrPC substrate. Since BHs contain many impurities, it is difficult to identify the co-factors using conventional PMCA. Thus, we developed a modified PMCA system using baculovirus and insect cell-derived recombinant PrP as a substrate (insect cell PMCA; iPMCA). We demonstrated that nucleic acids and glycosaminoglycans (GAGs) such as heparan sulfate (HS) or its analogue heparin (HP) are critical for PrPSc amplification in iPMCA. Of note, the addition of HS or HP restored the conversion efficiency in iPMCA under nucleic acid-depleted conditions. Moreover, the iPMCA products were infectious and preserved the strain properties of the input seed PrPSc. These data suggest that not only nucleic acids but also some GAGs play an important role in facilitating faithful replication of prions, at least in vitro.


Activation of both protein kinase A (PKA) type I and PKA type II isozymes is required for retinoid-induced maturation of acute promyelocytic leukemia cells.
Acute promyelocytic leukemia (APL) is characterized by granulopoietic differentiation arrest at the promyelocytic stage. In most cases, this defect can be overcome by treatment with all-trans-retinoic acid (ATRA), leading to complete clinical remission. Cyclic AMP signaling has a key role in retinoid treatment efficacy: it enhances ATRA-induced maturation in ATRA-sensitive APL cells (including NB4 cells) and restores it in some ATRA-resistant cells (including NB4-LR1 cells). We show that the two cell types express identical levels of the Cα catalytic subunit and comparable global cAMP-dependent protein kinase A (PKA) enzyme activity. However, the maturation-resistant NB4-LR1 cells have a PKA isozyme switch: compared with the NB4 cells, they have decreased content of the juxtanuclearly located PKA regulatory subunit IIα and PKA regulatory subunit IIβ, and a compensatory increase of the generally cytoplasmically distributed PKA-RIα. Furthermore, the PKA regulatory subunit II exists mainly in the less cAMP-responsive nonautophosphorylated state in the NB4-LR1 cells. By the use of isozyme-specific cAMP analog pairs, we show that both PKA-I and PKA-II must be activated to achieve maturation in NB4-LR1 as well as NB4 cells. Therefore, special attention should be paid to activating not only PKA-I but also PKA-II in attempts to enhance ATRA-induced APL maturation in a clinical setting.


Brain mechanisms mediating auditory attentional capture in humans.
The ability to detect and preferentially process salient auditory stimuli, even when irrelevant to a current task, is often critical for adaptive behavior. This stimulus-driven allocation of processing resources is known as "attentional capture." Here we used functional magnetic resonance imaging in humans to investigate brain activity and behavioral effects related to such auditory attentional capture. Participants searched a sequence of tones for a target tone that was shorter or longer than the nontarget tones. An irrelevant singleton feature in the tone sequence resulted in behavioral interference (attentional capture) and activation of parietal and prefrontal cortices only when the singleton was associated with a nontarget tone (nontarget singleton) and not when associated with a target tone (target singleton). In contrast, the presence (vs. absence) of a singleton feature in the sequence was associated with activation of frontal and temporal loci previously associated with auditory change detection. These results suggest that a ventral network involving superior temporal and inferior frontal cortices responds to acoustic variability, regardless of attentional significance, but a dorsal frontoparietal network responds only when a feature singleton captures attention.


Perspective: Sleep at the USA Science and Engineering Festival-A New Outreach for the American Academy of Sleep Medicine.
This April, the American Academy of Sleep Medicine (AASM) took part as exhibitors at the USA Science and Engineering Festival in Washington, DC. This was AASM's first time attending the festival which is the largest conference of its kind promoting and celebrating science and technology in the United States hosting 370,000 attendees including schoolchildren, educators, and the general public. The AASM's exhibit featured interactive games as well as materials aimed at the promotion of healthy sleep habits in all age groups. A few individuals presented with more specific questions and were provided education and directed to online resources approved by the Academy. It was apparent that many people were unaware of the field of sleep medicine and responded favorably to our presence. We hope our account of the experience helps inform thought on further direction the AASM takes in the realm of public outreach and education.


Nonstructural Protein 5A Impairs DNA Damage Repair: Implications for Hepatitis C Virus-Mediated Hepatocarcinogenesis.
RAD51-associated protein 1 (RAD51AP1) is a member of the multiprotein complexes postulated to carry out RAD51-mediated homologous recombination and DNA repair in mammalian cells. In the present study, we showed that hepatitis C virus (HCV) NS5A directly bound RAD51AP1 and increased the protein level of RAD51AP1 through modulation of the ubiquitin-proteasome pathway. We also demonstrated that RAD51AP1 protein levels were increased in the liver tissues of HCV-infected patients and NS5A-transgenic mice. Importantly, NS5A impaired DNA repair by disrupting the RAD51/RAD51AP1/UAF1 complex and rendered HCV-infected cells more sensitive to DNA damage. Silencing of RAD51AP1 expression resulted in a decrease of viral propagation. We further demonstrated that RAD51AP1 was involved in the assembly step of the HCV life cycle by protecting viral RNA. These data suggest that HCV exploits RAD51AP1 to promote viral propagation and thus that host DNA repair is compromised in HCV-infected cells. Overall, our findings provide mechanistic insight into the pathogenesis of HCV infection.IMPORTANCE Chronic infection with HCV is the leading cause of hepatocellular carcinoma (HCC). However, the molecular mechanisms underlying HCV-induced HCC are not fully understood. Here we demonstrate that the HCV NS5A protein physically interacts with RAD51AP1 and increases the RAD51AP1 protein level through modulation of the ubiquitin-proteasome pathway. HCV coopts host RAD51AP1 to protect viral RNA at an assembly step of the HCV life cycle. Note that the RAD51 protein accumulates in the cytoplasm of HCV-infected cells, and thus the RAD51/RAD51AP1/UAF1-mediated DNA damage repair system in the nucleus is compromised in HCV-infected cells. Our data may provide new insight into the molecular mechanisms of HCV-induced pathogenesis.


Comparison of immunoglobulin G subclass profiles induced by measles virus in vaccinated and naturally infected individuals.
A total of 258 human sera positive for measles antibodies were divided into four different groups: group 1 contained 54 sera from children after natural measles infection (immunoglobulin M [IgM] positive, early infection phase), group 2 contained 28 sera from children after measles vaccination (IgM positive, early infection phase), group 3 contained 100 sera from healthy adults (natural long-lasting immunity), and group 4 contained 76 sera from healthy children (postvaccinal long-lasting immunity). In the early phase of infection, the percent distributions of measles virus-specific IgG isotypes were similar between natural and postvaccinal immune responses. IgG1 and IgG4 were the dominant isotypes, with mean levels of detection of 100% (natural infection) and 100% (postvaccinal) for IgG1 and 96% (natural infection) and 92% (postvaccinal) for IgG4. In comparison, the IgG4 geometric mean titer (GMT) in the early phase of natural infection was significantly higher than the IgG4 GMT detected in the postvaccinal immune response (80 versus 13; 95% confidence interval). In the memory phase, IgG2 and IgG3 responses decreased significantly in both natural infection and postvaccinal groups, while IgG1 levels were maintained. In contrast, the IgG4 postvaccinal immune response decreased strongly in the memory phase, whereas IgG4 natural long-lasting immunity remained unchanged (9 versus 86%; P < 0.05). The results obtained suggest that IgG4 isotype could be used in the early phase of infection as a quantitative marker and in long-lasting immunity as a qualitative marker to differentiate between natural and postvaccinal immune responses.


Production of HanWoo (Bos taurus coreanae) fetuses following interbreed somatic cell nuclear transfer.
The possibility of producing HanWoo (Bos taurus coreanae) calves from transferable bovine embryos, obtained by interbreed nuclear transfer using Holstein cytoplasts and surrogates, was investigated. As donor nuclei, HanWoo fetal fibroblasts were used. Cells were induced into quiescence by serum deprivation for 4-7 days before nuclear transfer. In vitro matured Holstein oocytes were enucleated, and single donor cell was placed into the perivitelline space of enucleated oocyte. After reconstruction, the embryos were fused. activated and cultured. On day 7, the embryos that developed to the blastocyst stages were transferred into Holstein recipient cows on day 6 to 7 of estrous cycle (estrus=0). The reconstructed embryos were successfully fused (58.8%; 47/80), cleaved (91.5%; 43/47), and developed to blastocysts (29.8%; 14/47). Eleven blastocysts were transferred into 5 Holstein recipient cows. Two recipients were pregnant, confirmed by ultrasonography at day 60 of gestation. But, one of them was opened between on day 80 to 100 of pregnancy, and the other had a stillbirth on day 255. The stillborn calf was physically normal, and we couldn't find any evidence of anomaly. These results show that cells derived from HanWoo somatic cell lines can be reprogrammed by interbreed nuclear transfer and develop subsequently in vivo as well as in vitro.


Pharmacokinetic and pharmacodynamic evaluation of nano-fixed dose combination for hypertension.
The current study was planned to formulate, characterize and evaluate the pharmacokinetics, and pharmacodynamics of a novel 'NanoFDC' comprising hydrochlorothiazide, candesartan (CNDT) and amlodipine. The candidate drugs were loaded in poly(DL-lactide-co-glycolide) by emulsion-diffusion-evaporation method. The formulations were evaluated for their size, morphology, drug loading and in-vitro release individually. Repeat dose pharmacokinetic and pharmacodynamic study of the nano-fixed dose combination (FDC) was done in dexamethasone-induced animal model. The entrapment efficiencies ranged from 44 ± 2.1, 32.2 ± 4 and 40.5 ± 2.6% for amlodipine, hydrochlorothiazide and CNDT, respectively. The nanoparticles ranged in size from 164 to 215 nm. In-vitro release profile of the nanoformulation showed unto 90% release by day 7 in simulated gastric fluid and simulated intestinal fluid, respectively. In pharmacokinetic analysis a sustained-release for 7 days was observed in nano-FDC group. Once weekly oral dosing of nano-FDC of amlodipine, CNDT and hydrochlorothiazide provided adequate antihypertensive effect which was not statistically different from daily dosing of free drugs in dexamethasone-induced animal model. Once weekly oral dosing of nano-FDC of amlodipine, CNDT and hydrochlorothiazide provided adequate antihypertensive effect and was not statistically different from daily dosing of free drugs in dexamethasone-induced animal model. This study provides proof of concept of feasibility of once weekly dosing of a nano-FDC comprising three antihypertensive drugs, which can lead to significant improvement in patient adherence to therapy.


Septoplasty versus non-surgical management for nasal obstruction in adults with a deviated septum: economic evaluation alongside a randomized controlled trial.
For years, the benefits of septoplasty have been questioned. Due to the scarce and inconclusive literature, several National Health Service (NHS) Clinical Commissioning Groups in England decided to add septal surgery to their list of restricted procedures with low clinical value. Recently, evidence was obtained that septoplasty is actually more effective than non-surgical management for nasal obstruction in adults with a deviated septum. However, the relation between costs and effects of septoplasty remains unknown. We conducted an economic evaluation alongside an open, multicenter, pragmatic randomized controlled trial in two tertiary and 16 secondary referral hospitals in the Netherlands. Adults with nasal obstruction and a deviated septum were randomized to (1) septoplasty with or without concurrent turbinate surgery or (2) non-surgical management consisting of (a combination of) medical treatment and watchful waiting. Analyses were performed on an intention-to-treat basis. Single imputation nested in the bootstrap percentile method (using 5000 bootstrap replications) was performed to assess the effect of missing data. After 12 and 24 months, we assessed the incremental costs per quality-adjusted life year (QALY) gained from a healthcare and a societal perspective. A total of 203 adults were randomly assigned to septoplasty (N = 102) or non-surgical management (N = 101). After 12 months, the mean cost difference between septoplasty and non-surgical management using a healthcare or societal perspective was €1181 (95%CI €1038 to €1323) or €2192 per patient (95%CI €1714 to €2670), respectively. The mean QALY difference was 0.03 per patient (95%CI - 0.01 to 0.07). Incremental costs per QALY gained from a healthcare or societal perspective were €41,763 or €77,525, respectively. After 24 months, the mean cost difference between the two groups using a healthcare or societal perspective decreased to €936 (95%CI €719 to €1153) or €1671 per patient (95%CI €952 to €2390), respectively. The mean QALY difference increased to 0.05 per patient (95%CI - 0.03 to 0.14). Incremental costs per QALY gained from a healthcare or societal perspective became €17,374 or €31,024, respectively. Analyses of imputed data did not alter our findings. Depending on the selected perspective, cost-effectiveness threshold, and time horizon, septoplasty has the potential to be cost-effective. Despite considerable uncertainty, septoplasty seems to be cost-effective from a healthcare perspective, after 24 months against a threshold of €20,000 per QALY. From a societal perspective, septoplasty is not yet cost-effective after 24 months, but it comes closer to the cost-effectiveness threshold as time passes by. Nederlands Trial Register, NTR3868 (https://www.trialregister.nl/trial/3698). Prospectively registered on February 21, 2013.


[Role of ¹⁸F 2-fluoro-2-deoxyglucose positron emission tomography in upper gastrointestinal malignancies].
The ¹⁸F 2-fluoro-2-deoxyglucose (FDG) PET/CT scan is an imaging modality used in the management of patients with various types of malignancies. ¹⁸F-FDG PET/CT has demonstrated significant efficacy in the staging and detection of metastatic disease in malignancies of the gastrointestinal tract. The assessment of the metabolic response to chemotherapy and improvements of overall survivals in malignancies of esophagus and stomach has been demonstrated in several studies. This review focuses on the role of ¹⁸F-FDG PET/CT scan in staging, metastasis, predict of recurrence and assessment of metabolic response in malignancies of the upper gastrointestinal tract.


Association of circulating leptin with bone mineral density in males and females.
Leptin, a hormone produced by fat cells, possesses several physiological functions not limited to energy balance. Recently, leptin has been shown to inhibit bone formation through its action at the hypothalamus. In the present study, the authors explored the association of leptin with bone mineral density (BMD) in men and women according to menopausal status. Subjects consisted of 75 men, 51 premenopausal women and 63 postmenopausal women aged 20-80 years. All were healthy and did not take medication which may affect bone metabolism. BMD was measured at L2-4 and femoral neck by DEXA. Serum leptin concentrations were measured by radioimmunoassay. Serum leptin in males was independently related to BMD at L2-4 (r = -0.36, p < 0.05) and the femoral neck (r = -0.32, p < 0.05) in a multiple linear regression model with age, body mass index (BMI), serum free testosterone, estradiol and leptin as independent variables. In premenopausal women, serum leptin correlated negatively to L2-4 (r = -0.29, p < 0.01) and femoral neck BMD (r = -0.29, p < 0.05) independently of age and BMI. However, in postmenopausal women, no association of leptin with BMD was found after controlling for age and BMI. The authors concluded that circulating leptin is negatively associated with BMD in men and premenopausal women, but not in postmenopausal women. The negative associations found in both premenopausal women and men in the present study strengthen the notion that leptin may inhibit bone formation during the accumulation of bone mass early in life


Involvement of hepatocyte growth factor-induced epithelial-mesenchymal transition in human adenomyosis.
Adenomyosis is commonly believed to arise from the basalis endometrium. As an estromedin growth factor, hepatocyte growth factor (HGF) exhibits multiple functions in endometriosis, a disease commonly believed to arise from the functionalis endometrium. Here, we investigated the role of HGF in the occurrence of epithelial-mesenchymal transition (EMT) in adenomyosis. Full-thickness-biopsy specimens from endometrium to myometrium were collected after hysterectomy from women with and without adenomyosis. The relationship between HGF and E-cadherin (epithelial cell marker) and N-cadherin (mesenchymal cell markers) was examined at the gene and protein levels using endometrial epithelial cells (EECs) in culture and tissues by quantitative RT-PCR and immunohistochemistry. The gene and protein expressions of two transcriptional repressors of E-cadherin, SLUG and SNAIL, were examined using Ishikawa cells and in response to HGF and estrogen (E2). HGF down-regulated E-cadherin and up-regulated N-cadherin mRNA expression in EECs, and an inverse relationship in protein expression between HGF and E-cadherin was observed in basalis endometria derived from women with diffuse and focal adenomyosis. HGF induced morphological changes of EECs from a cobblestone-like appearance to spindle-shaped cells and promoted migration of EECs. Ishikawa cells exhibited up-regulation of SLUG/SNAIL gene expression in response to both HGF and E2 with an additive effect between them. HGF- and E2-promoted SLUG/SNAIL gene expression was significantly abrogated after pretreatment of cells with anti-HGF antibody or ICI 182720, an estrogen receptor antagonist. HGF may be involved in gland invagination deep into the myometrium by inducing EMT at the endo-myometrial junction in women with adenomyosis.


circRNA_100290 plays a role in oral cancer by functioning as a sponge of the miR-29 family.
Circular RNAs (circRNAs) represent a class of non-coding RNAs that are widely expressed in mammals. However, it is largely unknown about the function of human circRNAs and the roles of circRNAs in human oral squamous cell carcinomas (OSCC). Here we performed a comprehensive study of circRNAs in human OSCC using circRNA and mRNA microarrays, and identified many circRNAs that are differentially expressed between OSCC tissue and paired non-cancerous matched tissue. We further found a circRNA termed circRNA_100290 that served as a critical regulator in OSCC development. We discovered that circRNA_100290 was upregulated and co-expressed with CDK6 in OSCC tissue. Knockdown of circRNA_100290 decreased expression of CDK6 and inhibited proliferation of OSCC cell lines in vitro and in vivo. Via luciferase reporter assays, circRNA_100290 was observed to directly bind to miR-29 family members. Further EGFP/RFP reporter assays showed that CDK6 was the direct target of miR-29b. Taken together, we conclude that circRNA_100290 may function as a competing endogenous RNA to regulate CDK6 expression through sponging up miR-29b family members. Taken together, it indicates that circRNAs may exert regulatory functions in OSCC and may be a potential target for OSCC therapy.


Plasticity of chemoreceptor gene expression: Sensory and circuit inputs modulate state-dependent chemoreceptors.
Animals dramatically modify their chemosensory behaviors when starved, which could allow them to alter and optimize their food-search strategies. Dynamic changes in the gene expression of chemoreceptors may be a general mechanism underlying food and state-dependent changes in chemosensory behaviors. In our recent study,(1) we identified chemoreceptors in the ADL sensory neuron type of C. elegans that are modulated by feeding state and food availability. Here, we highllight our recent findings by which sensory inputs into ADL, neuronal outputs from ADL, and circuit inputs from the RMG interneuron, which is electrically connected to ADL, are required to regulate an ADL-expressed chemoreceptor. This sensory and circuit-mediated regulation of chemoreceptor gene expression is dependent on cell-autonomous pathways acting in ADL, e.g. KIN-29, DAF-2, OCR-2 and calcium signaling, and circuit inputs from RMG mediated by NPR-1. Based on these findings, we propose an intriguing but speculative feedback modulatory circuit mechanism by which sensory perception of food and internal state signals may be coupled to regulate ADL-expressed chemoreceptors, which may allow animals to precisely regulate and fine-tune their chemosensory neuron responses as a function of feeding state.


Spatial Mapping of Protein Adsorption on Mesoporous Silica Nanoparticles by Stochastic Optical Reconstruction Microscopy.
Exposure to biological fluid envelops a nanoparticle in layers of proteins and biomolecules, which has a profound impact on the nanoparticle's biological fate. Although the identities and amounts of the proteins in this "corona" have been thoroughly examined, the spatial arrangement of the proteins is unclear, a problem that is compounded on porous nanoparticles due to penetration of proteins within the porous network. To address this problem, we have developed a procedure based on information derived from stochastic optical reconstruction microscopy. We employed a mathematical model to reveal the penetration depth of several proteins within porous nanoparticles. Understanding protein penetration depth provides an explanation for the composition of the protein corona, aiding in the development of safe and effective particle-based therapies.


ICU visiting policies.
Head nurses from 78 ICUs in 37 northeast Ohio hospitals were interviewed about visiting policies. There was tremendous variation with regard to frequency and length of visits; 25% of these ICUs allowed only 2 visits/day, and 42% restricted visits to under 20 min. Most units rarely or never allowed children under 12 yr old to visit. Traditional rationales for restricted visiting are not supported by studies in the literature, nor are they consistent with current concepts of patients' rights. In an era where high technology and medicine by-the-numbers threaten to dehumanize patients, open visiting is an important part of the humanization process.


Sum Frequency Generation Vibrational Spectroscopy for Characterization of Buried Polymer Interfaces.
Sum frequency generation vibrational spectroscopy (SFG-VS) has become one of the most appealing technologies to characterize molecular structures at interfaces. In this focal point review, we focus on SFG-VS studies at buried polymer interfaces and review many of the recent publications in the field. We also cover the essential theoretical background of SFG-VS and discuss the experimental implementation of SFG-VS.


[The clinico-electroneuromyographic characteristics of acute inflammatory polyneuropathies in children].
Clinical electroneuromyographic examination (ENME) was carried out in 59 children at the age of 1-12 years with acute demyelinating polyneuropathies (PNP). The patients were divided into two groups, namely: 24 children with Guillain-Barré [correction of Giyenne-Barret] syndrome (the first) and 35 ones with diphtheritic PNP (the second group). The main clinical symptoms were investigated in each group. As a result of ENME investigation the axonal and demyelinating types of PNP's beginning were identified. They were distinguished both by severity of course and by rate of restoration. Diphtheritic PNP was characterized by demyelinating type of pathology rise with addition of axonal component in severe cases. The conduction through the additional nerves was investigated to prognose the development of bulbar disorders in PNP.


SARS-CoV2 RT-PCR assays: In vitro comparison of 4 WHO approved protocols on clinical specimens and its implications for real laboratory practice through variant emergence.
RT-PCR testing on nasopharyngeal swabs is a key component in the COVID-19 fighting, provided to use sensitive and specific SARS-CoV2 genome targets. In this study, we aimed to evaluate and to compare 4 widely used WHO approved RT-PCR protocols on real clinical specimens, to decrypt the reasons of the diverging results and to propose recommendations for the choice of the genome targets. 260 nasopharyngeal samples were randomly selected among the samples tested between Week-16, 2020 and week-16 2021, in the Institut Pasteur de Tunis, Tunisia, one of the referent laboratories of COVID-19 in Tunisia. All samples were tested by Charité, Berlin protocol (singleplex envelop (E) and singleplex RNA-dependent RNA polymerase (RdRp)), Hong Kong Universiy, China protocol (singleplex nucleoprotein (N) and singleplex Open reading frame Orf1b), commercial test DAAN Gene® (using the CDC China protocol), (triplex N, Orf1ab with internal control) and Institut Pasteur Paris protocol (IPP) (triplex IP2(nsp9) and IP4 (nsp12) with internal control). For IPP, a selection from samples positive by IP2 but negative with IP4 was re-tested by exactly the same protocol but this time in singleplex. New results were described and analyzed. In vitro analysis showed discordant results in 29.2% of cases (76 out of 260). The most discordant protocol is DAAN Gene® due to the false positive late signals with N target. Discordant results between the two protocol's targets are more frequent when viral load are low (high Ct values). Our results demonstrated that the multiplexing has worsened the sensitivity of the IP4 target. We provide concise recommendations for the choice of the genome targets, the interpretation of the results and the alarm signals which makes suspect a gene mutation.


Effects of ID-HWS1000 on the Perception of Bowel Activity and Microbiome in Subjects with Functional Constipation: A Randomized, Double-Blind Placebo-Controlled Study.
During constipation, indigestible foods, such as probiotics, prebiotics, and dietary fiber, may improve the bowel environment and activity. In this randomized, double-blind, and placebo-controlled study, the effects of ID-HWS1000, composed of Lactobacillus and Bifidobacterium species, xylooligosaccharide, and dietary fiber, were evaluated to determine whether it improves the perception of bowel activity or cause changes in the gut microbiome. Thirty Korean adults with "functional constipation" according to the Rome III criteria were randomly assigned to the following groups: 20 in the ID-HWS1000 group and 10 in the placebo group. ID-HWS1000 or the placebo was consumed by the participants for 4 weeks. To assess the changes in the perception of bowel activity, clinical data and gut microbiome analyses were conducted before and after the experiment. There were significant differences between the groups in the response to 9 of the 12 survey questions (the number and duration of bowel movements, amount of feces, number of irritant bowel movements, number of times bowel movements felt incomplete, shape of the feces, amount of gas in the gut, discomfort after defecation, and discomfort owing to constipation) (P < .05). There was a decrease in the proportion of Firmicutes (Ruminococcaceae and Lachnospiraceae) and an increase in Bacteroidetes (Bacteroidaceae) (P < .05). Moreover, ID-HWS1000 directly improved the discomfort associated with bowel movements, decreased the proportion of Lachnospiraceae, and increased the proportion of Bacteroidaceae. These results confirmed that ID-HWS1000 improves the perception of bowel activity and exerts positive changes in individuals with functional constipation.


Foot type analysis based on electronic pedobarography data in individuals with joint hypermobility syndrome/Ehlers-Danlos syndrome hypermobility type during upright standing.
Joint hypermobility syndrome/Ehlers-Danlos syndrome hypermobility type (JHS/EDS-HT) is a rheumatologic condition characterized by generalized joint hypermobility and musculoskeletal and nonmusculoskeletal findings related to congenital laxity of connective tissue. Because foot pain and other foot problems are reported to make daily life problematic to manage for individuals with JHS/EDS-HT, and thanks to the availability of modern technology, the aim of the present study was to quantitatively characterize foot type in individuals with JHS/EDS-HT during upright standing. Forty feet of 20 women with JHS/EDS-HT (mean ± SD age, 36.03 ± 14.01 years) were assessed clinically and with a pressure-sensitive mat during upright standing. Forty-five percent of feet had a high arch (pes cavus), 27.5% had a normal arch, and 27.5% had a low arch (pes planus or flatfoot). From a clinical perspective, the characterization of foot type in JHS/EDS-HT is important to identify, develop, and enhance the rehabilitative options. An understanding of the relationship between pes cavus and foot pain in these patients could, in fact, improve the clinical management of these patients.


Epidemiology, characteristics and treatment of patients with relapsing remitting multiple sclerosis and incidence of high disease activity: Real world evidence based on German claims data.
Multiple Sclerosis (MS) is a chronic inflammatory, immune mediated disease of the central nervous system, with Relapsing Remitting MS (RRMS) being the most common type. Within the last years, the status of high disease activity (HDA) has become increasingly important for clinical decisions. Nevertheless, little is known about the incidence, the characteristics, and the current treatment of patients with RRMS and HDA in Germany. Therefore, this study aims to estimate the incidence of HDA in a German RRMS patient population, to characterize this population and to describe current drug treatment routines and further healthcare utilization of these patients. A claims data analyses has been conducted, using a sample of the InGef Research Database that comprises data of approximately four million insured persons from around 70 German statutory health insurances (SHI). The study was conducted in a retrospective cohort design, including the years 2012-2016. Identification of RRMS population based on ICD-10 code (ICD-10-GM: G35.1). For identification of HDA, criteria from other studies as well as expert opinions have been used. Information on incidence, characteristics and current treatment of patients with RRMS and HDA was considered. The overall HDA incidence within the RRMS population was 8.5% for 2016. It was highest for the age group of 0-19 years (29.4% women, 33.3% men) and lowest for the age group of ≥ 50 years (4.3% women, 5.6% men). Mean age of patients with RRMS and incident HDA was 38.4 years (SD: 11.8) and women accounted for 67.8%. Analyses of drug utilization showed that 82.4% received at least one disease-modifying drug (DMD) in 2016. A percentage of 49.8% of patients received drugs for relapse therapy. A share of 55% of RRMS patients with HDA had at least one hospitalization with a mean length of stay of 13.9 days (SD: 18.3 days) in 2016. The average number of outpatient physician contacts was 28.1 (SD: 14.0). This study based on representative Germany-wide claims data from the SHI showed a high incidence of HDA especially within the young RRMS population. Future research should consider HDA as an important criterion for the quality of care for MS patients.


Personalization of Medical Treatment Decisions: Simplifying Complex Models while Maintaining Patient Health Outcomes.
Personalizing medical treatments based on patient-specific risks and preferences can improve patient health. However, models to support personalized treatment decisions are often complex and difficult to interpret, limiting their clinical application. We present a new method, using machine learning to create meta-models, for simplifying complex models for personalizing medical treatment decisions. We consider simple interpretable models, interpretable ensemble models, and noninterpretable ensemble models. We use variable selection with a penalty for patient-specific risks and/or preferences that are difficult, risky, or costly to obtain. We interpret the meta-models to the extent permitted by their model architectures. We illustrate our method by applying it to simplify a previously developed model for personalized selection of antipsychotic drugs for patients with schizophrenia. The best simplified interpretable, interpretable ensemble, and noninterpretable ensemble models contained at most half the number of patient-specific risks and preferences compared with the original model. The simplified models achieved 60.5% (95% credible interval [crI]: 55.2-65.4), 60.8% (95% crI: 55.5-65.7), and 83.8% (95% crI: 80.8-86.6), respectively, of the net health benefit of the original model (quality-adjusted life-years gained). Important variables in all models were similar and made intuitive sense. Computation time for the meta-models was orders of magnitude less than for the original model. The simplified models share the limitations of the original model (e.g., potential biases). Our meta-modeling method is disease- and model- agnostic and can be used to simplify complex models for personalization, allowing for variable selection in addition to improved model interpretability and computational performance. Simplified models may be more likely to be adopted in clinical settings and can help improve equity in patient outcomes.


Role for glia in synaptogenesis.
Nearly one-half of the cells in a human brain are astrocytes, but the function of these little cells remains a great mystery. Astrocytes form an intimate association with synapses throughout the adult CNS, where they help regulate ion and neurotransmitter concentrations. Recent in vitro studies, however, have found that astrocytes also exert powerful control over the number of CNS synapses that form, are essential for postsynaptic function, and are required for synaptic stability and maintenance. Moreover, recent studies increasingly implicate astrocytes in vivo as participants in activity-dependent structural and functional synaptic changes throughout the nervous system. Taken together, these data force us to rethink the role of glia. We propose that astrocytes should not be viewed primarily as support cells, but rather as cells that actively control the structural and functional plasticity of synapses in developing and adult organisms.


Hes genes and neurogenin regulate non-neural versus neural fate specification in the dorsal telencephalic midline.
The choroid plexus in the brain is unique because it is a non-neural secretory tissue. It secretes the cerebrospinal fluid and functions as a blood-brain barrier, but the precise mechanism of specification of this non-neural tissue has not yet been determined. Using mouse embryos and lineage-tracing analysis, we found that the prospective choroid plexus region initially gives rise to Cajal-Retzius cells, specialized neurons that guide neuronal migration. Inactivation of the bHLH repressor genes Hes1, Hes3 and Hes5 upregulated expression of the proneural gene neurogenin 2 (Ngn2) and prematurely depleted Bmp-expressing progenitor cells, leading to enhanced formation of Cajal-Retzius cells and complete loss of choroid plexus epithelial cells. Overexpression of Ngn2 had similar effects. These data indicate that Hes genes promote specification of the fate of choroid plexus epithelial cells rather than the fate of Cajal-Retzius cells by antagonizing Ngn2 in the dorsal telencephalic midline region, and thus this study has identified a novel role for bHLH genes in the process of deciding which cells will have a non-neural versus a neural fate.


Solid-phase synthesis of lipidated peptides.
Characteristic partial structures of lipidated proteins embodying different lipid groups as well as additional fluorescent tags or a maleimide for coupling to proteins can be synthesized readily by means of a new solid-phase technique employing the oxidative cleavage of the hydrazide linker as well as on-resin farnesylation and palmitoylation after appropriate deprotection of cysteine thiol groups as the key steps.


Smoking, pregnancy and the fetus.
Smoking may influence a women's reproductive ability by having adverse effects on fertility, the early embryo and the fetus. The ability to become pregnant and to maintain the pregnancy long enough for it to be clinically detected is reduced in smokers. Smoking can adversely influence several of the crucial steps within the reproductive process required for achieving a pregnancy. Despite the increased miscarriage rate in women that smoke there is no evidence to suggest that smoking can cause fetal abnormalities. The chemical composition of cigarette smoke is more closely related to reduction in fetal growth than the number of cigarettes smoked. In smokers, the placenta appears remarkably normal. The effects of smoking could be due to one or more of the hundreds of different chemical substances in tobacco smoke, but apart from carbon monoxide and nicotine, little is known about the effects of other toxins. Nicotine can adversely affect uterine and placental blood flow by causing constriction of the blood vessels. It can also affect both the maternal and fetal blood pressure and heart rate, but its action on the fetal cardiovascular system does not appear to be dependent on its effect on placental perfusion.


Regulation of human B lymphocyte responsiveness.
Regulation of human B cell responses is a complex process involving the activities of a variety of cells and cytokines. There are important differences between the regulation of human and murine B lymphocytes, especially with regard to the action of IL2 and IL4. In man, IL2 appears to play a central role in regulating B cell activation, proliferation and differentiation, whereas IL4 plays a regulatory role by inhibiting initial B cell activation. Above and beyond providing regulatory cytokines, activated human T cells can also directly stimulate B cell differentiation by a process that requires cellular contact. A better understanding of the nature of signal transduction during interactions between B cells and activated T cells and provided by the various cytokines should provide additional insight into the regulation of B cell function in man.


Naked suicide.
Suicide attempts and completions by individuals while naked remain unexplored, both by clinicians and in the professional literature. Nakedness at the time of the incident conveys important information about an individual's mental state. Motivations for attempting or completing suicide while naked are discussed. Nakedness during a suicide attempt is presumptive evidence of high risk for suicide completion. Deliberate self-harm, without intent to die, is not usually inflicted while naked. The clinical and forensic implications of naked suicide are explored. In completed suicides, the state of dress, including partial undress and nakedness may provide important clues in a psychological autopsy. Forensic psychiatrists and treating clinicians who encounter naked suicides are afforded a unique opportunity to advance our knowledge. Letters to the editor and case reports would stimulate discourse about this little understood phenomenon.


Rapid quantitative analysis of adulterant Lonicera species in preparations of Lonicerae Japonicae Flos.
Lonicerae Japonicae Flos is often adulterated with Lonicerae Flos, which is derived from the other four Lonicera species, in both the crude drug and Lonicerae Japonicae Flos preparations. We proposed a methodology for the quantitative analysis of adulterant Lonicerae Flos in Lonicerae Japonicae Flos preparations. Taking macranthoidins A, B, dipsacoside B (saponins), sweroside (iridoids), and luteolin-7-O-d-glucoside (flavonoids) as markers, a method of ultra high performance liquid chromatography with triple quadrupole mass spectrometry was employed to determine their amounts in Lonicerae Flos, Lonicerae Japonicae Flos, and Lonicerae Japonicae Flos preparations. The proportion of adulterant Lonicerae Flos in Lonicerae Japonicae Flos preparations was estimated based on the saponin contents of Lonicerae Japonicae Flos and Lonicerae Flos. All analytes separated under isocratic elution in 12 min with acceptable linearity, precision, repeatability, and accuracy. Lonicerae Japonicae Flos was easily distinguished from Lonicerae Flos by the total amount of saponins (0.067 and > 45.8 mg/g for Lonicerae Japonicae Flos and Lonicerae Flos, respectively). Eighteen of twenty one Lonicerae Japonicae Flos preparation samples were adulterated with Lonicerae Flos in proportions of 11.3-100%. The developed ultra high performance liquid chromatography with triple quadrupole mass spectrometry method could be used for the identification of Lonicerae Japonicae Flos and the four species of Lonicerae Flos and for the analysis of Lonicerae Japonicae Flos preparations adulterated with Lonicerae Flos.


Individual- and area-level socioeconomic inequalities in diabetes mellitus in Saskatchewan between 2007 and 2012: a cross-sectional analysis.
Improving our understanding of social inequalities may improve prevention and treatment efforts for diabetes mellitus. We examined the association between individual- and area-level socioeconomic measures and physician-diagnosed diabetes in Saskatchewan over time. In this cross-sectional study, we linked health administrative data with individual-level socioeconomic data from the Canadian Community Health Survey and area-level data from the 2006 Canadian census. We used general linear mixed-models regression to analyze the effect of each factor, controlling for geographic and demographic measures. Area-level deprivation was associated with medically diagnosed type 2 diabetes mellitus after adjustment for the individual-level factors of age, sex, household income and education. Individuals residing in areas ranked in the least deprived quintile had a lower likelihood of diabetes than those in the most deprived quintile (odds ratio 0.40, 95% confidence interval 0.18-0.88). However, this disparity existed only in urban areas. This result may reflect less pronounced health inequalities in rural areas, greater socioeconomic heterogeneity, larger geographic units or some combination of these factors. Individual- and area-level socioeconomic factors were associated with the likelihood of medically diagnosed diabetes; however, the strength of this association varied between urban and rural communities. Acknowledgement of area-level deprivation as a modifiable risk factor related to the prevalence of diabetes is important in the development of effective interventions for urban, but not rural, areas.


Identifying School-Based Factors that Predict Employment Outcomes for Transition-Age Youth with Autism Spectrum Disorder.
There is a growing need to provide appropriate services to help students with autism spectrum disorder (ASD) transition to employment. Limited research has investigated what aspects of support should be prioritized when preparing youth with ASD for employment. By conducting structural equation modeling using a nationally-representative dataset on high school students receiving special education services (NLTS-2), this study examined the malleable predictors of employment during the transition and developed a model to examine the relationships between predictors and employment outcomes. The findings suggested two pathways for youth with ASD. For youth with higher daily functioning skills (DFS), academic performance mediated the relationship between parent participation and employment. For youth with lower DFS, school-based transition supports was the key mediator.


Correlation between reduction in macular vessel density and frequency of intravitreal ranibizumab for macular oedema in eyes with branch retinal vein occlusion.
To determine whether there is a significant correlation between the reduction in macular vessel density and the recurrence of macular oedema in eyes with branch retinal vein occlusion (BRVO). We reviewed the medical records of 27 eyes with macular oedema associated with BRVO. All eyes had received pro re nata intravitreal ranibizumab (IVR) injection. A reinjection of ranibizumab was performed when the central foveal thickness was ≥300 µm. At 12 months, patients without additional IVR for the last 4 months were placed in the resolved group, otherwise they were placed in the recurrence group. The macular vessel density in 3×3 mm area centred on the fovea was determined by optical coherence tomography angiography. At 12 months, 11 of the 27 (40.7%) eyes were placed in the resolved group and the other 16 (59.3%) eyes were placed in the recurrence group. The mean macular vessel density reduction in the resolved group was 17.60%±7.88% and that in the recurrence group was 8.12%±7.48% (P=0.0042). The mean number of IVR injections in the resolved group was 2.1±1.1 and that in the recurrence group was 5.1±1.2 (P<0.0001). In all eyes, multivariate analysis showed that the reduction in macular vessel density was significantly and negatively correlated with the number of IVR injections (β=-0.6746, P=0.0028). A reduction in the macular vessel density was significantly correlated with the number of recurrences of macular oedema associated with BRVO.


Electrical impedance properties of carbon nanotube composite electrodes for chemical and biosensor.
Electrical impedance properties of different type of carbon nanotubes based bulk electrodes have been investigated to develop chemical and biosensors. The bulk composite electrodes were fabricated with single-wall and multi-wall carbon nanotubes involving ionic conducting host polymer, Nafion, by using traditional solution-casting techniques. Under the various amounts of buffer solution, resistance and capacitance of the electrodes were measured with LCR meter and their characteristics due to ionic conducting host polymer were investigated by means of electrokinetic analysis. The capacitance values showed drastic change while the resistances only changed within few percent ranges. Electrical impedance measurement provided rapid and simple sensing mechanism to develop chemical sensor and biosensors with bulk nano electrodes.


EGFR Mutation and 11q13 Amplification Are Potential Predictive Biomarkers for Immunotherapy in Head and Neck Squamous Cell Carcinoma.
Head and neck squamous cell carcinoma (HNSCC) is one of the most common malignant cancers. The treatment of HNSCC remains challenging despite recent progress in targeted therapies and immunotherapy. Research on predictive biomarkers in clinical settings is urgently needed. Next-generation sequencing analysis was performed on tumor samples from 121 patients with recurrent or metastatic HNSCC underwent sequencing analysis. Clinicopathological information was collected, and the clinical outcomes were assessed. Progression-free survival (PFS) was estimated using the Kaplan-Meier method and cox regression model was used to conduct multivariate analysis. Fisher's exact tests were used to calculate clinical benefit. A p value of less than 0.05 was designated as significant (p < 0.05). Chromosome 11q13 amplification (CCND1, FGF3, FGF4, and FGF19) and EGFR mutations were significantly associated with decreased PFS and no clinical benefits after treatment with a programmed death 1 (PD-1) inhibitor. The same results were found in the combined positive score (CPS) ≥ 1 subgroup. In patients who were treated with an EGFR antibody instead of a PD-1 inhibitor, a significant difference in PFS and clinical benefits was only observed between patients with CPS ≥ 1 and CPS < 1. Chromosome 11q13 amplification and EGFR mutations were negatively correlated with anti-PD-1 therapy. These markers may serve as potential predictive biomarkers to identify patients for whom immunotherapy may be unsuitable.


Arsenic binding to human metallothionein.
The number of reported cases of chronic arsenic poisoning is on the rise throughout the world, making the study of the long-term effects of arsenic critical. As(3+) binds readily to biological thiols, including mammalian metallothionein (MT), which is an ubiquitous sulfur-rich metalloprotein known to coordinate a wide range of metals. The two-domain mammalian protein binds divalent metals (M) into two metal-thiolate clusters with stoichiometries of M(3)S(cys9) (beta) and M(4)S(cys11) (alpha). We report that As(3+) binds with stoichiometries of As(3)S(cys9) (beta) and As(3)S(cys11) (alpha) to the recombinant human metallothionein (rhMT) isoform 1a protein. Further, we report the complete kinetic analysis of the saturation reactions of the separate alpha and beta domains of rhMT with As(3+). Speciation in the metalation reactions was determined using time- and temperature-resolved electrospray ionization mass spectrometry. The binding reaction of As(3+) to the alpha and beta MT domains is shown to be noncooperative and involves three sequential, bimolecular metalation steps. The analyses allow for the first time the complete simulation of the experimental data for the stepwise metalation reaction of MT showing the relative concentrations of the metal-free, apo MT and each of the As-MT intermediate species as a function of time and temperature. At room temperature (298 K) and pH 3.5, the individual rate constants for the first, second, and third As(3+) binding to apo-alphaMT are 5.5, 6.3, and 3.9 M(-)(1) s(-)(1) and for apo-betaMT the constants are 3.6, 2.0, and 0.6 M(-)(1) s(-)(1). The activation energy for formation of As(1)-H(6)-betaMT is 32 kJ mol(-)(1), for As(2)-H(3)-betaMT it is 35 kJ mol(-)(1), for As(3)-betaMT it is 29 kJ mol(-)(1), for As(1)-H(8)-alphaMT it is 33 kJ mol(-)(1), for As(2)-H(5)-alphaMT it is 29 kJ mol(-)(1), and for As(3)-H(2)-alphaMT it is 23 kJ mol(-)(1).


Influence of pH and time on organic substance release from a model dental composite: a fluorescence spectrophotometry and gas chromatography/mass spectrometry analysis.
In this study we assessed the influence of pH and time on the degradation and elution of organic substances from the composite resin material, Z-100. To accomplish this, fluorescence spectrophotometry was evaluated as an appropriate technique for the identification of six organic substances (methacrylic acid, methyl methacrylate, hydroquinone, ethylene glycol dimethacrylate, triethylene glycol dimethacrylate and 4,4'-isopropylidenediphenol) that were eluted from resin composite material stored for 24 h or 6 months at pH 4.0, 6.0 or 8.0. In addition, complementary analyses (solid-phase microextraction/gas chromatography/mass spectrometry) were carried out to identify and quantify the substances. The main substances leached from the resin composite were methacrylic acid, triethylene glycol dimethacrylate and hydroquinone. It was concluded that fluorescence spectrophotometry seems to be a suitable, non-destructive technique for the qualitative analysis of eluted organic components. Critical combinations of time and pH allowed the elution of several organic substances, predominantly methacrylic acid, triethylene glycol dimethacrylate and hydroquinone, from the model resin composite, Z-100.


Effect of continuous positive airway pressure treatment on serum adiponectin level and mean arterial pressure in male patients with obstructive sleep apnea syndrome.
Recent research suggested that obstructive sleep apnea syndrome (OSAS) might be independently associated with hypoadiponectinemia, which was linked to some complications of OSAS, such as hypertension, diabetes, etc. This study was conducted to investigate the effect of continuous positive airway pressure (CPAP) treatment on changes of both serum adiponectin levels and mean arterial pressure and their possible links in male OSAS patients. Twenty-three adult male patients with moderate-to-severe OSAS but without obesity, coronary heart disease and diabetes were recruited. Their blood samples were collected and morning mean arterial pressure (MAP) was measured before CPAP treatment and on day 3, 7, 14 of CPAP treatment respectively. The serum adiponectin concentration was tested with radioimmunoassay. Compared with the serum adiponectin level before CPAP treatment, no significant change was found in OSAS patients on day 3 and day 7 of CPAP treatment (P > 0.05). It was not until day 14 of CPAP treatment did a significant elevation in serum adiponectin level occur (P < 0.01). Meanwhile, the MAP showed no statistically significant difference among its levels before CPAP, on day 3 and day 7 of CPAP treatment (P > 0.05). However, on day 14 of CPAP treatment, a significantly lower MAP than that obtained before treatment was observed (P < 0.05). CPAP treatment can gradually reverse hypoadiponectinemia and reduce MAP in OSAS patients. Hypoadiponectinemia might be involved in the pathogenesis of OSAS-mediated hypertension.


Plaque-infiltrating T lymphocytes in patients with carotid atherosclerosis: an insight into the cellular mechanisms associated to plaque destabilization.
Inflammation plays a crucial role in the progression of atherosclerotic plaques. The aim of the present study was to investigate phenotypic and functional characteristics of plaque-infiltrating T lymphocytes associated with a complicated phenotype of carotid atherosclerotic lesions. Atherosclerotic plaques were obtained from 17 patients undergoing carotid endarterectomy and cultured to isolate infiltrating T lymphocytes. Blood samples were obtained from patients and from 20 sex- and age-matched healthy subjects. The presence of lymphocytes (CD3+ cells) within atherosclerotic plaques was determined by immunohistochemistry. Phenotypic characteristics and intracellular cytokine expression of plaque-infiltrating and circulating T lymphocytes were determined by flow cytometry. Cytokine levels in supernatants from infiltrating T cell cultures were evaluated by enzyme-linked immunosorbent assay. A higher number of CD3+ cells was detected in complicated than in uncomplicated plaques. Complicated plaques had higher percentages of tumor necrosis factor (TNF)-α- and interferon (IFN)-γ- positive cells than uncomplicated ones, especially in CD4+ subpopulation. In patients the percentages of TNF-α-positive cells were higher in infiltrating than in circulating lymphocyte samples. Intracellular TNF-α, IFN-γ, interleukin (IL)-4 and IL-10 expression resulted higher in circulating lymphocyte samples from patients than in those from healthy subjects. Supernatants of infiltrating T cell cultures from complicated plaques showed higher levels of TNF-α and lower levels of IL-4 than those from uncomplicated plaques. Our data provide new information on the presence of increased percentages of pro-inflammatory T lymphocytes in complicated plaques with respect to uncomplicated ones and support the concept of the key role played by activated T cells in the progression of atherosclerotic lesions.


Hotspot of Crimean-Congo Hemorrhagic Fever Virus Seropositivity in Wildlife, Northeastern Spain.
We conducted a serosurvey for Crimean-Congo hemorrhagic fever virus antibodies in various wildlife species in Catalonia, northeastern Spain. We detected high seroprevalence in southern Catalonia, close to the Ebro Delta wetland, a key stopover for birds migrating from Africa. Our findings could indicate that competent virus vectors are present in the region.


Design and construction of the high-speed optoelectronic memory system demonstrator.
The high-speed optoelectronic memory system project is concerned with the reduction of latency within multiprocessor computer systems (a key problem) by the use of optoelectronics and associated packaging technologies. System demonstrators have been constructed to enable the evaluation of the technologies in terms of manufacturability. The system combines fiber, free space, and planar integrated optical waveguide technologies to augment the electronic memory and the processor components. Modeling and simulation techniques were developed toward the analysis and design of board-integrated waveguide transmission characteristics and optical interfacing. We describe the fabrication, assembly, and simulation of the major components within the system.


[Spontaneous rupture of infected renal cyst presenting sudden onset of right flank distension: a case report].
A 21-year-old woman was referred to our hospital for sudden onset of right flank distension. Ultrasonography, abdominal CT scan and MRI revealed a giant cyst 20 x 11 x 10 cm on the right kidney. Percutaneous puncture of the lesion was performed and fluid was drained. The fluid was yellowish and puriform. Gradually, the drainage fluid creatinine concentration increased. Cystography revealed communication between the lesion and the pyelocalyceal system. The patient underwent drainage for 25 days, causing the cyst to almost fully collapse. No recurrence has been detected in the follow-up for about 1 year. This case is the 38th case of the rupture of a renal cyst reported in the Japanese literature. We reviewed 37 cases of rupture of renal cysts previously reported in Japan.


Chronic constipation in childhood: a longitudinal study of 186 patients.
One hundred eighty-six patients with chronic constipation and soiling were seen between 1975 and 1982. On follow-up, 47% had resolution of symptoms, with another 36% having their soiling controlled with either continuous or intermittent laxatives. Thirteen clinical features were analyzed in relation to outcome. The only predictor of poor outcome was the presence of soiling (p = 0.003), while the presence of abdominal pain correlated well with cure (p = 0.007). Fifty-five percent of patients had a positive family history for constipation. Significant psychological problems were present in 20% of patients; however, these did not appear to affect the outcome. Constipation was uncommon over the age of 12 years, and there was no evidence of the development of laxative dependence. These findings suggest that chronic idiopathic constipation of childhood may be a constitutional condition which tends to resolve with age.


Possible control of transcript levels by chlorophyll precursors in Chlamydomonas.
Steady-state mRNA levels of the three nuclear genes cab1, rbcS1 and rbcS2 (coding for the light-harvesting chlorophyll-binding protein (LHCP) and the small subunit of ribulose 1,5-bisphosphate carboxylase, respectively) and of the two plastid-encoded genes rbcL and psaA2 (coding for the large subunit of the carboxylase and a member of the P700 chlorophyll a protein, respectively) have been investigated in synchronized Chlamydomonas cells in response to light and inhibitors interfering with chlorophyll synthesis. The accumulation of cab1, rbcS1 and psaA2 transcripts is light-dependent, whereas transcripts from rbcS2 and rbcL genes are present in high amounts in the light and in the dark. Dioxoheptanoic acid, an inhibitor blocking chlorophyll synthesis prior to porphyrin formation, does not affect the accumulation of all five mRNAs. However, inhibition of chlorophyll synthesis by incubating cells with dipyridyl, cycloheximide or nitrogen promotes the accumulation of porphyrin compounds, but specifically prevents the accumulation of light-dependent transcripts. Although functionally unrelated, these inhibitors are known to block an Fe-dependent oxygenase, which is involved in the formation of the isocyclic ring in the chlorophyll molecule. The data are explained as a control by chlorophyll precursors over the accumulation of light-dependent transcripts.


Late Postoperative Complications in Laparoscopic Sleeve Gastrectomy (LVSG) Versus Laparoscopic Roux-en-y Gastric Bypass (LRYGB): Meta-analysis and Systematic Review.
Laparoscopic Roux-en-Y gastric bypass (LRYGB) and laparoscopic vertical sleeve gastrectomy (LVSG), have been proposed as cost-effective strategies to manage obesity-related chronic disease. The objectives of this meta-analysis and systematic review were to analyze the "late postoperative complication rate (>30 days)" for these 2 procedures. Randomized controlled trials (RCTs) published between 2000 and 2015 comparing the late complication rates, that is, >30 days following LVSG and LRYGB in adult population (ie, 16 y and above) were selected from PubMed, Medline, Embase, Science Citation Index, Current Contents, and the Cochrane database. The outcome variables analyzed included mortality rate, major and minor complications, and interventions required for their management and readmission rates. Random effects model was used to calculate the effect size of both binary and continuous data. Heterogeneity among the outcome variables of these trials was determined by the Cochran Q statistic and I index. The meta-analysis was prepared in accordance with the Preferred Reporting of Systematic Reviews and Meta-Analyses guidelines. Six RCTs involving a total of 685 patients (LVSG, n=345; LRYGB, n=340) reported late major complications. A nonstatistical reduction in relative odds favoring the LVSG procedure was observed [odds ratio (OR), 0.64; 95% confidence interval (CI), 0.21-1.97; P=0.4]. Four RCTs representing 408 patients (LVSG, n=208; LRYGB, n=200) reported late minor complications. A nonstatistically significant reduction of 36% in relative odds favoring the LVSG procedure was observed (OR, 0.64; 95% CI, 0.28-1.47; P=0.3). A 37% relative reduction in odds was observed in favor of the LVSG for the need for additional interventions to manage late postoperative complications that did not reach statistical significance (OR, 0.63; 95% CI, 0.19-2.05; P=0.4). No study specifically reported readmissions required for the management of late complication. This meta-analysis and systematic review of RCTs shows that the development of late (major and minor) complications is similar between LVSG and LRYGB procedures, 6 months to 3 years postoperatively, and they do not lead to higher readmission rate or reoperation rate for either procedure. However longer-term surveillance is required to accurately describe the patterns of late complications in these patients.


Transcystic approach to laparoscopic common bile duct exploration.
One-stage laparoscopic management for common bile duct stones in patients with gallbladder stones has gained wide acceptance. We developed a novel technique using a transcystic approach for common bile duct exploration as an alternative to the existing procedures. From April 2010 to June 2012, 9 consecutive patients diagnosed with cholelithiasis and common bile duct stones were enrolled in this study. The main inclusion criteria included no upper abdominal surgical history and the presence of a stone measuring <5 mm. After the gallbladder was dissected free from the liver connections in a retrograde fashion, the fundus of the gallbladder was extracted via the port incision in the right epigastrium. The choledochoscope was inserted into the gallbladder through the small opening in the fundus of the gallbladder extracorporeally and was advanced toward the common bile duct via the cystic duct under the guidance of both laparoscopic imaging and endoscopic imaging. After stones were retrieved under direct choledochoscopic vision, a drainage tube was placed in the subhepatic space. Of 9 patients, 7 had successful transcystic common bile duct stone clearance. A narrow cystic duct and the unfavorable anatomy of the junction of the cystic duct and common bile duct resulted in losing access to the common bile duct. No bile leakage, hemobilia, or pancreatitis occurred. Wound infection occurred in 2 patients. Transient epigastric colic pain occurred in 2 patients and was relieved by use of anisodamine. A transient increase in the amylase level was observed in 3 patients. Short-term follow-up did not show any recurrence of common bile duct stones. Our novel transcystic approach to laparoscopic common bile duct exploration is feasible and efficient.


Four of a Kind: A Complete Collection of ADP-Ribosylated Histidine Isosteres Using Cu(I)- and Ru(II)-Catalyzed Click Chemistry.
Adenosine diphosphate ribosylation (ADP-ribosylation) is a crucial post-translational modification involved in important regulatory mechanisms of numerous cellular pathways including histone maintenance and DNA damage repair. To study this modification, well-defined ADP-ribosylated peptides, proteins, and close analogues thereof have been invaluable tools. Recently, proteomics studies have revealed histidine residues to be ADP-ribosylated. We describe here the synthesis of a complete set of triazole-isosteres of ADP-ribosylated histidine to serve as probes for ADP-ribosylating biomachinery. By exploiting Cu(I)- and Ru(II)-catalyzed click chemistry between a propargylglycine building block and an α- or β-configured azidoribose, we have successfully assembled the α- and β-configured 1,4- and 1,5-triazoles, mimicking N(τ)- and N(π)-ADP-ribosylated histidine, respectively. The ribosylated building blocks could be incorporated into a peptide sequence using standard solid-phase peptide synthesis and transformed on resin into the ADP-ribosylated fragments to provide a total of four ADP-ribosyl triazole conjugates, which were evaluated for their chemical and enzymatic stability. The 1,5-triazole analogues mimicking the N(π)-substituted histidines proved susceptible to base-induced epimerization and the ADP-ribosyl α-1,5-triazole linkage could be cleaved by the (ADP-ribosyl)hydrolase ARH3.


Object detection and basic-level categorization: sometimes you know it is there before you know what it is.
A tight temporal coupling between object detection (is an object there?) and object categorization (what kind of object is it?) has recently been reported (Grill-Spector & Kanwisher, 2005), suggesting that image segmentation into different objects and categorization of those objects at the basic level may be the very same mechanism. In the present work, we decoupled the time course of detection and categorization through two task manipulations. First, inverted objects were categorized significantly less accurately than upright objects across a range of image presentation durations, but no significant effect on object detection was observed. Second, systematically degrading stimuli affected categorization significantly more than object detection. The time course of object detection and object categorization can be selectively manipulated. They are not intrinsically linked. As soon as you know an object is there, you do not necessarily know what it is.


Design of experiments to optimize an in vitro cast to predict human nasal drug deposition.
Previous studies showed nasal spray in vitro tests cannot predict in vivo deposition, pharmacokinetics, or pharmacodynamics. This challenge makes it difficult to assess deposition achieved with new technologies delivering to the therapeutically beneficial posterior nasal cavity. In this study, we determined best parameters for using a regionally divided nasal cast to predict deposition. Our study used a model suspension and a design of experiments to produce repeatable deposition results that mimic nasal deposition patterns of nasal suspensions from the literature. The seven-section (the nozzle locator, nasal vestibule, front turbinate, rear turbinate, olfactory region, nasopharynx, and throat filter) nylon nasal cast was based on computed tomography images of healthy humans. It was coated with a glycerol/Brij-35 solution to mimic mucus. After assembling and orienting, airflow was applied and nasal spray containing a model suspension was sprayed. After disassembling the cast, drug depositing in each section was assayed by HPLC. The success criteria for optimal settings were based on nine in vivo studies in the literature. The design of experiments included exploratory and half factorial screening experiments to identify variables affecting deposition (angles, airflow, and airflow time), optimization experiments, and then repeatability and reproducibility experiments. We found tilt angle and airflow time after actuation affected deposition the most. The optimized settings were flow rate of 16 L/min, postactuation flow time of 12 sec, a tilt angle of 23°, nozzle angles of 0°, and actuation speed of 5 cm/sec. Neither cast nor operator caused significant variation of results. We determined cast parameters to produce results resembling suspension nasal sprays in the literature. The results were repeatable and unaffected by operator or cast. These nasal spray parameters could be used to assess deposition from new devices or formulations. For human deposition studies using radiolabeled formulations, this cast could show that radiolabel deposition represents drug deposition. Our methods could also be used to optimize settings for other casts.


Patients' expectations for the functional and psychosocial outcomes of orbital decompression surgery for thyroid eye disease: a qualitative study.
Patients with appearance-altering conditions may be dissatisfied with the outcomes of reconstructive surgery due to unmet expectations. This study explored patients' expectations of orbital decompression surgery for thyroid eye disease (TED) and whether these were met. Semi-structured interviews were conducted at two times: (1) in the weeks after patients were listed for decompression surgery and before surgery; (2) up to 12 months after surgery. Thematic analysis was performed for each time point, to identify themes within the data. Fourteen adults with TED were interviewed prior to surgery and five were followed up after surgery. Thematic analyses found: (1) Prior to surgery, patients had formed expectations through online information about the procedure, consultations with physicians, the impact TED had on their lives, and speaking to relevant others. Patients had specific expectations about the procedure, the recovery, post-operative appearance and post-operative vision. (2) After surgery, patients generally felt their appearance and well-being had improved. However, dissatisfaction was linked to unanticipated specific aspects of surgical care, recovery, or appearance. Dissatisfaction can arise from unmet expectations for the outcomes of reconstructive surgery. Physicians should be aware of the processes by which patients form expectations, for example different types and quality of online information. Ensuring that preoperative expectations are realistic could enhance satisfaction after surgery.


An Investigation of Influencing Factors on Practical Sub-Diffraction-Limit Focusing of Planar Super-Oscillation Lenses.
Planar super-oscillation lenses (SOLs) can fulfill super-resolution focusing and nanoscopic imaging in the far field without the contribution of evanescent waves. Nevertheless, the existing deviations between the design and experimental results have been seldomly investigated, leaving the practical applications of SOLs unpredictable and uncontrollable. In this paper, some application-oriented issues are taken into consideration, such as the inevitable fabrication errors and the size effect of the designed SOLs, with the aim of providing an engineering reference to elaborately customize the demanded focusing light field. It turned out that a thicker structural film makes the focal spots enlarged, while the sloped sidewalls just weaken the intensity of the focal hotspot. Furthermore, the focal lengths are diminished with the decrease of device size, while the focal spots are enlarged. This research will promote the wide-spread applications of SOLs for sub-diffraction-limit far-field focusing in the areas of nanoscopy and high-density optical storage.


Prevention of glucocorticoid-induced osteoporosis.
Patients taking low-dose glucocorticoids for 3 months or more are at risk for rapid bone loss. The loss may be as high as 15% in the first 3 to 6 months of treatment, and can lead to osteoporotic fractures, with an increase in morbidity and mortality. This article discusses an evidence-based approach to the prevention of glucocorticoid-induced osteoporosis.


Plasma FGF21 levels in obese patients undergoing energy-restricted diets or bariatric surgery: a marker of metabolic stress?
Fibroblast growth factor 21 (FGF21) has been suggested to be an endocrine signal of nutritional status and an active regulator of metabolism. However, there is no agreement on the effect of weight-loss therapies on circulating levels of FGF21 in humans. To assess FGF21 circulating levels in adiposity excess and after different weight-loss strategies prescribed in five different groups from four independent centers. Body composition, ketosis, insulin sensitivity and FGF21 were evaluated in 181 excess body weight and 14 normal-weight subjects. From the excess body weight patients, two independent groups (discovery cohort; n=20 and validation cohort; n=28) undertook a very low-calorie ketogenic (VLCK) diet, a third group followed a low-calorie (LC) diet (n=84) and other two groups underwent bariatric surgery (discovery cohort; n=24 and validation cohort; n=25). The follow-up was 4 to 6 or 12 months, respectively. FGF21 levels were higher in excess body weight patients than in normal-weight subjects. The energy-restriction therapy to lose weight induced a significant decrease, with respect to baseline, in circulating levels of FGF21 (VLCK: -62.5 pg ml-1 or -14.8 pg ml-1 and LC diet: -67.9 pg ml-1). There were no differences in FGF21 levels between both energy-restriction treatments. On the contrary, after bariatric surgery morbidly obese patients showed a significant increase in FGF21, especially 1 month after surgery (148.8 pg ml-1 higher than baseline). The FGF21 differential changes occur concomitantly with a non-induced ketosis situation (0.66±0.56 mm) in bariatric surgery, and an improvement in adiposity and insulin sensitivity induced by the three therapies. FGF21 levels were reduced after energy-restricted treatments and severely increased after bariatric surgery, independently of the weight reduction magnitude, insulin sensitivity or ketosis. Therefore, FGF21 appears to be a marker of severe nutritional stress.


[Audiovisual stimulation in children with severely limited motor function: does it improve their quality of life?].
Children with neurological diseases that impose severe limitations on their mobility have a deficient quality of life (QL). To study whether the QL of such patients improves with the application of a programme of audiovisual stimulation. A prospective study was conducted on nine children, six of whom were males (mean age: 42.6 ± 28.6 months), with severely limited mobility and who had been hospitalised for long periods. Two audiovisual stimulation programmes were produced and applied, together with videos, by means of a specially designed structure. The stimulus was applied twice a day for 10 minutes over 20 days. The first ten days the stimulus was carried out in a passive manner and the second block of ten days it was performed with the guidance of the observer. Biological, behavioural and cognitive variables were recorded and an adapted QL survey was applied. Three cases of spinal muscular atrophy, two of congenital muscular dystrophy, two of myopathy and two with other conditions were diagnosed. Eight patients completed the follow-up. From the baseline point of view, they presented a regular QL (7.2 ± 1.7 points; median: 7.0; range: 6-10), which improved to good on finishing the therapy (9.4 ± 1.2 points; median: 9.0; range: 8-11), with an intra-individual difference of 2.1 ± 1.6 (median: 2.5; range: -1 to 4; CI 95% = 0.83-3.42; p = 0.006). Improved cognition and a favourable perception in the caregivers were detected. No changes took place in the biological or behavioural variables. Audiovisual stimulation can be used to enhance the quality of life of children with severely limited mobility.