Single-cell RNA-seq analysis of the brainstem of mutant SOD1 mice reveals perturbed cell types and pathways of amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease in which motor neurons throughout the brain and spinal cord progressively degenerate resulting in muscle atrophy, paralysis and death. Recent studies using animal models of ALS implicate multiple cell-types (e.g., astrocytes and microglia) in ALS pathogenesis in the spinal motor systems. To ascertain cellular vulnerability and cell-type specific mechanisms of ALS in the brainstem that orchestrates oral-motor functions, we conducted parallel single cell RNA sequencing (scRNA-seq) analysis using the high-throughput Drop-seq method. We isolated 1894 and 3199 cells from the brainstem of wildtype and mutant SOD1 symptomatic mice respectively, at postnatal day 100. We recovered major known cell types and neuronal subpopulations, such as interneurons and motor neurons, and trigeminal ganglion (TG) peripheral sensory neurons, as well as, previously uncharacterized interneuron subtypes. We found that the majority of the cell types displayed transcriptomic alterations in ALS mice. Differentially expressed genes (DEGs) of individual cell populations revealed cell-type specific alterations in numerous pathways, including previously known ALS pathways such as inflammation (in microglia), stress response (ependymal and an uncharacterized cell population), neurogenesis (astrocytes, oligodendrocytes, neurons), synapse organization and transmission (microglia, oligodendrocyte precursor cells, and neuronal subtypes), and mitochondrial function (uncharacterized cell populations). Other cell-type specific processes altered in SOD1 mutant brainstem include those from motor neurons (axon regeneration, voltage-gated sodium and potassium channels underlying excitability, potassium ion transport), trigeminal sensory neurons (detection of temperature stimulus involved in sensory perception), and cellular response to toxic substances (uncharacterized cell populations). DEGs consistently altered across cell types (e.g., Malat1), as well as cell-type specific DEGs, were identified. Importantly, DEGs from various cell types overlapped with known ALS genes from the literature and with top hits from an existing human ALS genome-wide association study (GWAS), implicating the potential cell types in which the ALS genes function with ALS pathogenesis. Our molecular investigation at single cell resolution provides comprehensive insights into the cell types, genes and pathways altered in the brainstem in a widely used ALS mouse model.


Compartmentalized regulation of macrophage arachidonic acid metabolism.
We show that downregulation of arachidonic acid (20:4) metabolism which occurs following i.p. injection of C. parvum can occur in a single, localized macrophage population, and is therefore unlikely to be mediated solely by a systemic factor.


Hepatoprotective and antioxidant effects of Hygrophila auriculata (K. Schum) Heine Acanthaceae root extract.
Hygrophila auriculata (K. Schum) Heine (syn. Asteracantha longifolia Nees, Acanthaceae) was widely used in the Indian systems of medicine for the treatment of various liver ailments. The hepatoprotective activity of the aqueous extract of the roots was studied on CCl(4)-induced liver toxicity in rats. The activity was assessed by monitoring the various liver function tests, viz. alanine transaminase, aspartate transaminase (AST), alkaline phosphatase (ALP), total protein and total bilirubin. Furthermore, hepatic tissues were subjected to histopathological studies. The root extract was also studied for its in vitro antioxidant activity using ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods. The extract exhibited significant hepatoprotective and antioxidant activities.


Uncovering the Role of Crystal Phase in Determining Nonvolatile Flash Memory Device Performance Fabricated from MoTe2-Based 2D van der Waals Heterostructures.
Although the crystal phase of two-dimensional (2D) transition metal dichalcogenides (TMDs) has been proven to play an essential role in fabricating high-performance electronic devices in the past decade, its effect on the performance of 2D material-based flash memory devices still remains unclear. Here, we report the exploration of the effect of MoTe2 in different phases as the charge-trapping layer on the performance of 2D van der Waals (vdW) heterostructure-based flash memory devices, where a metallic 1T'-MoTe2 or semiconducting 2H-MoTe2 nanoflake is used as the floating gate. By conducting comprehensive measurements on the two kinds of vdW heterostructure-based devices, the memory device based on MoS2/h-BN/1T'-MoTe2 presents much better performance, including a larger memory window, faster switching speed (100 ns), and higher extinction ratio (107), than that of the device based on the MoS2/h-BN/2H-MoTe2 heterostructure. Moreover, the device based on the MoS2/h-BN/1T'-MoTe2 heterostructure also shows a long cycle (>1200 cycles) and retention (>3000 s) stability. Our study clearly demonstrates that the crystal phase of 2D TMDs has a significant impact on the performance of nonvolatile flash memory devices based on 2D vdW heterostructures, which paves the way for the fabrication of future high-performance memory devices based on 2D materials.


Satellite Glial Cells: Morphology, functional heterogeneity, and role in pain.
Neurons in the somatic, sympathetic, and parasympathetic ganglia are surrounded by envelopes consisting of satellite glial cells (SGCs). Recently, it has become clear that SGCs are highly altered after nerve injury, which influences neuronal excitability and, consequently, the development and maintenance of pain in different animal models of chronic pain. However, the exact mechanism underlying chronic pain is not fully understood yet because it is assumed that SGCs in different ganglia share many common peculiarities, making the process complex. Here, we review recent data on morphological and functional heterogeneity and changes in SGCs in various pain conditions and their role in response to injury. More research is required to decipher the role of SGCs in diseases, such as chronic pain, neuropathology, and neurodegenerative diseases.


Potential of veg2 blastomeres to induce endoderm differentiation in sea urchin embryos.
Two different modes of gastrulation in sea urchin embryos have been reported. The first mode, reported in Hemicentrotus pulcherrimus and some other species, consists of two phases: a primary and a secondary invagination. The second mode involves gastrulation with a continuous convolution of cells near the blastopore; this mode has been reported to occur in the embryos of the sand dollar, Scaphechinus mirabilis. The rudimentary gut is comprised of fewer cells in the embryos of the former species than in the latter. We assumed that the differences in gastrulation modes could be related to the different potentials of the veg2 layer to induce endoderm differentiation in the upper layer. In the present study, we produced chimeric embryos consisting of an animal cap recombined with veg2 layer blastomere(s) to compare the inductive effect of the veg2 layer and/or the blastomere(s) in H. pulcherrimus and S. mirabilis embryos. Our results showed that the inductive effect of the veg2 layer is stronger in S. mirabilis embryos than in H. pulcherrimus embryos. Moreover, it was suggested that the difference in the strength of inductive effects of veg2 layers is related to the difference in gastrulation modes.


Germination of conidia of Aspergillus niger is accompanied by major changes in RNA profiles.
The transcriptome of conidia of Aspergillus niger was analysed during the first 8 h of germination. Dormant conidia started to grow isotropically two h after inoculation in liquid medium. Isotropic growth changed to polarised growth after 6 h, which coincided with one round of mitosis. Dormant conidia contained transcripts from 4 626 genes. The number of genes with transcripts decreased to 3 557 after 2 h of germination, after which an increase was observed with 4 780 expressed genes 8 h after inoculation. The RNA composition of dormant conidia was substantially different than all the subsequent stages of germination. The correlation coefficient between the RNA profiles of 0 h and 8 h was 0.46. They were between 0.76-0.93 when profiles of 2, 4 and 6 h were compared with that of 8 h. Dormant conidia were characterised by high levels of transcripts of genes involved in the formation of protecting components such as trehalose, mannitol, protective proteins (e.g. heat shock proteins and catalase). Transcripts belonging to the Functional Gene Categories (FunCat) protein synthesis, cell cycle and DNA processing and respiration were over-represented in the up-regulated genes at 2 h, whereas metabolism and cell cycle and DNA processing were over-represented in the up-regulated genes at 4 h. At 6 h and 8 h no functional gene classes were over- or under-represented in the differentially expressed genes. Taken together, it is concluded that the transcriptome of conidia changes dramatically during the first two h and that initiation of protein synthesis and respiration are important during early stages of germination.


[Assessment of quality of life for home ventilated patients with Duchenne muscular dystrophy].
A social and psychological survey was conducted for patients with DMD submitted to home mechanical ventilation for more than one year. Thirty six were tracheostomized and 16 were using non invasive ventilation. Fifty two patients were recruited: 36 were tracheostomized and 16 were using non invasive ventilation. Mean age was 25 +/- 5 years. In the two groups: 1- the main disagreement was air leaking and cutaneous erosions. 2- when present, headache, dyspnea, sleep troubles and general fatigue were improved by mechanical ventilation. 3- ventilation was considered as improving health but with an increasing of dependencies. 4- Majors disappointment are sexual life (70 percent) and physical status (40 percent) but patients spent more than half time with positive feeling (92 percent). There was no difference between satisfaction evaluation and type of ventilation. Home ventilated patients with DMD have positive assessment of satisfaction. Despite technical disagreement as air leaking, patients feel an improvement of their life and advise other DMD to use early home ventilation.


Rational design, synthesis, and characterization of novel inhibitors for human beta1,4-galactosyltransferase.
An affinity labeling reagent, uridine 5'-(6-amino-{2-[(7-bromomethyl-2-naphthyl)methoxycarbonylmethoxy]ethoxy}acetyl-6-deoxy-alpha-D-galactopyranosyl) diphosphate (1a), was designed on the basis of 3D docking simulation and synthesized to investigate the functional role of Trp310 residue located in the small loop near the active site of human recombinant galactosyltransferase (betaGalT-1). Mass spectrometric analysis revealed that the Trp310 residue of betaGalT1 can be selectively modified with the naphthylmethyl group of compound 1a at the C-3 position of the indole ring. This result motivated us to synthesize novel uridine-5'-diphosphogalactose (UDP-Gal) analogues as candidates for mechanism-based inhibitors for betaGalT-1. We found that uridine 5'-(6-O-[10-(2-naphthyl)-3,6,9-trioxadecanyl]-alpha-d-galactopyranosyl) diphosphate (2) is the strongest inhibitor (K(i) = 1.86 microM) against UDP-Gal (Km = 4.91 microM) among compounds reported previously. A cold spray ionization time-of-flight mass spectrometry study demonstrated that the complex of this inhibitor and betaGalT-1 cannot interact with an acceptor substrate in the presence of Mn2+.


Spinal cord trauma in man: studies of phosphorylated neurofilament and ubiquitin expression.
Changes in neuronal proteins are a major feature of the neurodegenerative disorders, but the biological significance of these changes is not understood. We describe alterations in neuronal proteins as a result of surgical tractotomy on the human spinal cord at 3, 5, 7, 14 and 72 days after operation. Antibodies to phosphorylated neurofilaments labelled neuronal cell bodies in the nucleus dorsalis and anterior horns. Axonal swellings showed positive staining for ubiquitin, but ubiquitinated neuronal inclusions were not evident. The presence of phosphorylated neurofilaments in the neuronal cell bodies and of ubiquitination of axonal swellings may be indicators of interrupted axonal transport. The changes found in intracellular proteins in neurons after trauma do not resemble those found in motor neuron disease, suggesting that the intraneuronal ubiquitinated inclusions may have a unique significance in the cytopathology of that disorder.


Long-range distance measurements in proteins at physiological temperatures using saturation recovery EPR spectroscopy.
Site-directed spin labeling in combination with EPR is a powerful method for providing distances on the nm scale in biological systems. The most popular strategy, double electron-electron resonance (DEER), is carried out at cryogenic temperatures (50-80 K) to increase the short spin-spin relaxation time (T2) upon which the technique relies. A challenge is to measure long-range distances (20-60 Å) in proteins near physiological temperatures. Toward this goal we are investigating an alternative approach based on the distance-dependent enhancement of spin-lattice relaxation rate (T1(-1)) of a nitroxide spin label by a paramagnetic metal. With a commonly used nitroxide side chain (R1) and Cu(2+), it has been found that interspin distances ≤25 Å can be determined in this way (Jun et al. Biochemistry 2006, 45, 11666). Here, the upper limit of the accessible distance is extended to ≈40 Å using spin labels with long T1, a high-affinity 5-residue Cu(2+) binding loop inserted into the protein sequence, and pulsed saturation recovery to measure relaxation enhancement. Time-domain Cu(2+) electron paramagnetic resonance, quantum mechanical calculations, and molecular dynamics simulations provide information on the structure and geometry of the Cu(2+) loop and indicate that the metal ion is well-localized in the protein. An important aspect of these studies is that both Cu(2+)/nitroxide DEER at cryogenic temperatures and T1 relaxation measurements at room temperature can be carried out on the same sample, allowing both validation of the relaxation method and assessment of the effect of freezing on protein structure.


In Vitro Analysis of the Role of Schwann Cells on Axonal Degeneration and Regeneration Using Sensory Neurons from Dorsal Root Ganglia.
Sensory neurons from dorsal root ganglion efficiently regenerate after peripheral nerve injuries. These neurons are widely used as a model system to study degenerative mechanisms of the soma and axons, as well as regenerative axonal growth in the peripheral nervous system. This chapter describes techniques associated to the study of axonal degeneration and regeneration using explant cultures of dorsal root ganglion sensory neurons in vitro in the presence or absence of Schwann cells. Schwann cells are extremely important due to their involvement in tissue clearance during axonal degeneration as well as their known pro-regenerative effect during regeneration in the peripheral nervous system. We describe methods to induce and study axonal degeneration triggered by axotomy (mechanical separation of the axon from its soma) and treatment with vinblastine (which blocks axonal transport), which constitute clinically relevant mechanical and toxic models of axonal degeneration. In addition, we describe three different methods to evaluate axonal regeneration using quantitative methods. These protocols constitute a valuable tool to analyze in vitro mechanisms associated to axonal degeneration and regeneration of sensory neurons and the role of Schwann cells in these processes.


Exteriorization and control of a retained embolization coil in the carotid artery--a case report.
Advances in endovascular technology have enabled the development of complex techniques for the treatment of vascular conditions. Not surprisingly, the modern vascular surgeon will likely encounter unusual complications and will need to formulate plans for their management. In the current case report, the vascular surgery service was consulted to assist in the management of a retained embolization coil in the carotid artery. Relevant aspects of detachable coils are discussed and the successful management of this potentially hazardous complication is described.


A fast 4D cone beam CT reconstruction method based on the OSC-TV algorithm.
Four-dimensional cone beam computed tomography allows for temporally resolved imaging with useful applications in radiotherapy, but raises particular challenges in terms of image quality and computation time. The purpose of this work is to develop a fast and accurate 4D algorithm by adapting a GPU-accelerated ordered subsets convex algorithm (OSC), combined with the total variation minimization regularization technique (TV). Different initialization schemes were studied to adapt the OSC-TV algorithm to 4D reconstruction: each respiratory phase was initialized either with a 3D reconstruction or a blank image. Reconstruction algorithms were tested on a dynamic numerical phantom and on a clinical dataset. 4D iterations were implemented for a cluster of 8 GPUs. All developed methods allowed for an adequate visualization of the respiratory movement and compared favorably to the McKinnon-Bates and adaptive steepest descent projection onto convex sets algorithms, while the 4D reconstructions initialized from a prior 3D reconstruction led to better overall image quality. The most suitable adaptation of OSC-TV to 4D CBCT was found to be a combination of a prior FDK reconstruction and a 4D OSC-TV reconstruction with a reconstruction time of 4.5 minutes. This relatively short reconstruction time could facilitate a clinical use.


A role for virtue in unifying the 'knowledge' and 'caring' discourses in nursing theory.
A critical examination of contemporary nursing theory suggests that two distinct discourses coexist within this field. On the one hand, proponents of the 'knowledge discourse' argue that nurses should drop the 'virtue script' and focus on the scientific and technical aspects of their work. On the other hand, proponents of the 'caring discourse' promote a view of nursing that embodies humanistic qualities such as compassion, empathy and mutuality. In view of this, we suggest a way to reconcile both discourses despite the fact that they appear to be at odds theoretically and practically. To that end, we argue that nursing theory must give a prominent role to the Aristotelian conception of virtue, and we offer an account that includes both character and intellectual virtues. This account allows for a focus on moral competence but also accommodates the demands for discipline-specific knowledge. Our account incorporates the caring discourse by suggesting a way for individuals to cultivate the conditions within themselves that make 'caring in nursing' possible, while the knowledge discourse is accommodated via the acquisition of the intellectual virtues. The process for achieving both these ends is the same: an intention to consistently develop, hone and exercise certain character traits over time.


[Healing of bronchial anastomoses in single and double pulmonary transplantations].
Bilateral single lung transplantation (BSLT) (the procedure of choice for double lung replacement) and single lung transplantation (SLT) have at present a low incidence of anastomotic problems. In our experience, we have recorded no death related to anastomotic problems and 4 complications occurred for 33 anastomoses with a follow up of at least 2 months. A stent was temporarily inserted for two patients. The decrease of anastomotic complications is confirmed in the literature. It is one of the reasons why the number of SLT and BSLT performed all over the world increases rapidily to the detriment of heart lung transplantation.


A green light-absorbing phycoerythrin is present in the high-light-adapted marine cyanobacterium Prochlorococcus sp. MED4.
In the high-light-adapted unicellular marine cyanobacterium Prochlorococcus sp. MED4 the cpeB gene is the only gene coding for a structural phycobiliprotein. The absence of any other phycoerythrin gene in the fully sequenced genome of this organism, the previous inability to detect a gene product, and the mutation of two out of four cysteine residues, normally involved in binding chromophores, suggested that MED4-cpeB might not code for a functional protein. Here, transcription of MED4-cpeB at a low level was detected and the transcriptional start site was mapped. Enrichment of the protein identified phycoerythrobilin as its sole chromophore in vivo, which was confirmed by chromophorylation assays in vitro using the recombinant protein. Phycourobilin is the major chromophore in low-light-adapted Prochlorococcus ecotypes such as strain SS120. Therefore, spectrally tuned phycoerythrins are a characteristic feature of distinct Prochlorococcus ecotypes. Further in vitro mutagenesis experiments replacing one or both cysteines C61R/C82S by arginine or serine, respectively, revealed that only Cys82 is required for chromophore binding. Thus, an unusual green light-absorbing phycoerythrin evolved in the high-light-adapted ecotypes of Prochlorococcus, which potentially serves as a photoreceptor.


Influence of immunosuppression in patients with severe acute respiratory distress syndrome on veno-venous extracorporeal membrane oxygenation therapy.
Prognosis of patients suffering from acute respiratory distress syndrome (ARDS) is poor. This is especially true for immunosuppressed patients. It is controverisal whether these patients should receive veno-venous extracorporeal membrane oxygenation (VV ECMO) while evidence on this topic is sparse. We report retrospective data of a single-center registry of patients with severe ARDS requiring ECMO support between October 2010 and June 2019. Patients were analyzed by their status of immunosuppression. ECMO weaning success and hospital survival were analyzed before and after propensity score matching (PSM). Moreover, ventilator free days (VFD) were compared. A total of 288 patients were analyzed (age 55 years, 67% male), 88 (31%) presented with immunosuppression. Survival rates were lower in immunosuppressed patients (27% vs. 53%, P < .001 and 27% vs. 48% after PSM, P = .006). VFD (60 days) were lower for patients with immunosuppression (11.9 vs. 22.4, P < .001), and immunosuppression was an independent predictor for mortality in multivariate analysis. Hospital survival was 20%, 14%, 35%, and 46% for patients with oncological malignancies, solid organ transplantation, autoimmune diseases, and HIV, respectively. In this analysis immunosuppression was an independent predictor for mortality. However, there were major differences in the weaning and survival rates between the etiologies of immunosuppression which should be considered in decision making.


DNA bases attack by reactive metabolites produced during carbon tetrachloride biotransformation and promotion of liver microsomal lipid peroxidation.
Free radicals produced during carbon tetrachloride biotransformation and the promoted lipid peroxidation process of liver microsomal lipids are able to attack the DNA bases guanine, cytosine and thymine to give at least three altered bases. They were identified as 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua); 5-hydroxycytosine (5OHCyt) and 5-hydroxymethyluracil (5OHMeU). No adenine adducts were detected. If these altered bases were formed during carbon tetrachloride poisoning and were not adequately repaired before cell replication, serious permanent consequences for liver cell DNA could be expected and this might be somehow involved in the liver carcinogenic effects of the haloalkane.


Gene organization and promoter function for CC chemokine receptor 5 (CCR5).
CC chemokine receptor 5 (CCR5) functions physiologically as a receptor for the leukocyte chemoattractants macrophage inflammatory protein-1alpha, macrophage inflammatory protein-1beta, and RANTES, and functions pathologically as a key cell entry coreceptor for HIV-1. The factors that regulate CCR5 expression may be useful therapeutic targets for HIV-1 infection. To identify nuclear regulatory factors, we have located and functionally characterized the CCR5 gene promoter. The gene consists of two exons separated by a 1.9-kb intron. Exon 1 contains 43 bp of the 5'-untranslated region; exon 2 contains 11 bp of the 5'-untranslated region and the complete open reading frame. Primer extension analysis identified two adjacent transcriptional start points (tsp) that map to the first 2 bp found in the longest known CCR5 cDNA sequence. A TATA box is present 31 bp upstream from the first tsp. CCR5 mRNA was detected constitutively in both primary human myeloid and lymphoid cells by Northern blot hybridization. Consistent with this, transcription of a chloramphenicol acetyltransferase reporter gene was constitutively activated in both transiently transfected myeloid and lymphoid cell lines by the 80-bp gene fragment located immediately upstream of the tsp. Deletion analysis located a strong silencer element between nucleotides -244 and -80, and a strong enhancer element between -486 and -244. These results suggest that the gene region between -486 and -1 may regulate the expression of CCR5 in monocyte/macrophages and T lymphocytes.


A new multiplex real-time polymerase chain reaction assay for the diagnosis of periprosthetic joint infection.
A new multiplex real-time polymerase chain reaction (PCR) assay was developed to detect methicillin-resistant Staphylococcus (MRS) and to distinguish between gram-positive and gram-negative bacteria. In this study, we validated the sensitivity and specificity of this assay with periprosthetic joint infections (PJIs) and evaluated the utility of PCR for culture-negative PJI. Forty-five samples from 23 infectious PJI cases and 106 samples from 64 non-infectious control cases were analyzed by real-time PCR using a LightCycler Nano® system. Twenty-eight clinical samples, comprising bacteria of known species isolated consecutively in the microbiological laboratory of our hospital, were used to determine the spectrum of bacterial species that could be detected using the new multiplex primers and probes. The sensitivity and specificity of the MRS- and universal-PCR assays were 92% and 99%, and 91% and 88%, respectively. Twenty-eight species of clinically isolated bacteria were detected using this method and the concordance rate for the identification of gram-positive or gram-negative organisms was 96%. Eight samples were identified as PCR-positive despite a culture-negative result. This novel multiplex real-time PCR system has acceptable sensitivity and specificity and several advantages; therefore, it has potential use for the diagnosis of PJIs, particularly in culture-negative cases.


Alcohol use and the development of periodontal pockets: An 11-year follow-up study.
This study investigates whether alcohol use predicts the periodontal pocket development over an 11-year follow-up period. The study participants' periodontal condition was examined both in the Health 2000 and 2011 Surveys (BRIF8901). Participants were aged 30 to 65 years, dentate, periodontally healthy, and did not have diabetes or rheumatoid arthritis in 2000. Periodontal outcome measures in 2011 were the number of teeth with deepened (≥4 mm) periodontal pockets and the presence of deepened periodontal pockets. The exposure in 2000 was alcohol use (amount [g/week], frequency [any alcohol or different beverages]). Incidence rate ratios (IRRs) with 95% confidence intervals (CI) were estimated using negative binomial regression model and Poisson regression model with a robust variance estimator. Overall, the amount of alcohol use or use over the risk limit in 2000 was inconsistently associated with the development of periodontal pockets (IRRs varied from 0.6 to 1.0). The frequency of alcohol use (any alcohol or different beverages) had an inconsistent association with the presence of periodontal pockets (IRRs varied from 0.5 to 1.2) while there was an inverse association with the number of teeth with periodontal pockets. Among smokers, there were no clear associations between any of the exposures and either of the outcomes. The same was found among non-smokers except an inverse association was found between frequency of alcohol use and the number of teeth with periodontal pockets. Alcohol use was not consistently associated with the periodontal pocket development over a period of 11 years.


Determination of active components in Cynanchum chinense R. Br. by capillary electrophoresis.
A method for the determination of 7-O-alpha-l-rhamnopyranosyl-kaempferol-3-O-beta-d-glucopyranoside (GL) and 7-O-alpha-l-rhamnopyranosyl-kaempferol-3-O-alpha-l-rhamnopyranoside (RH) in the traditional Chinese herb Cynanchum chinense R. Br. by capillary electrophoresis has been developed. With botate buffer (30 mmol/L, pH 9.50) as running buffer and an applied voltage of 20 kV, the compounds were completely separated within 6 min and detected at UV 254 nm. The correlation coefficients of the calibration curves for GL and RH were 0.9990 and 0.9992, respectively, over the concentration ranges (15.0-1000.0 and 12.0-1000.0 microg/mL), and the recoveries were from 91.4 to 107.1%.


Derivatives of usnic acid cause cytostatic effect in Caco-2 cells.
Usnic acid has anti-cancer activity, however, low solubility and toxicity limit the potential. To investigate biological activity of usnic acid derivatives, enantiopure derivatives were synthesised by reacting usnic acid with ethylenediamine, which yielded one dimer product ((+)-1), and two tetra cyclic compounds ((+)-2 and (-)-2). The products were characterised with NMR, and evaluated in vitro in human colon cancer cell line Caco-2 by cell count, phase-contrast microscopy, MTT-assay, measurement of DNA content and cell cycle distribution. All compounds tested showed cytostatic effect in Caco-2 cells, but each compound had a distinct cellular effect. Compound (+)-1 showed anti-proliferative activity by increasing the percentage of cells in S-phase with 25% compared to the control. Compounds (+)-2 and (-)-2 induced paraptosis, but only compound (+)-2 modulated cell cycle distribution by accumulating cells in G2/M-phase by 47% and reduced DNA content by 60%. All compounds express interesting cellular and potential anti-proliferative activity.


Rheumatoid arthritis in a patient with compound heterozygous variants in the COL11A2 gene and progressive hearing loss: A case report.
Collagen type XI alpha 2 chain is a component of type XI collagen and is expressed in various tissues including articular cartilage and tectorial membrane of the cochlea. Variants in the COL11A2 gene, which encodes collagen type XI alpha 2 chain, has been reported to cause hearing loss and has been associated with osteoarthritis and ossification of the posterior longitudinal ligament of the spine. Despite the importance of type XI collagen in the joints, association of rheumatoid arthritis (RA) with COL11A2 has not been reported. The patient is a 60-year-old female, born to Japanese parents of no known consanguinity. She had progressive hearing loss since childhood. Her father also had progressive hearing loss before middle age. She developed joint pain in the knees and the hips in her forties. When she was 56, she developed polyarthritis. Rheumatoid factor and anti-CCP antibodies were positive. She was diagnosed with osteoarthritis and RA. Whole exome analysis detected 2 rare variants, c.4201C>T, p.(Arg1401Trp) and c4265C>T, p.(Pro1422Leu), in the COL11A2 gene (NM_080680.2). Whole genome analysis with a long insert size confirmed 2 variants that are in trans. She received a cochlear implant, which improved her hearing. She was treated with methotrexate, golimumab, tocilizumab, and upadacitinib with partial responses for her RA. We herein report a patient with RA with compound heterozygous variants in the COL11A2 gene. Autoantibodies against type XI collagen are detected in the sera of patients with RA, suggesting the possibility that type XI collagen may be involved in the pathogenesis of RA as an autoantigen. The hearing loss and osteoarthritis in this patient may be due to the compound heterozygous variants in the COL11A2 gene, and the conformational changes induced by the variants may have changed the immunogenicity of type XI collagen, leading to the development of RA.


Connexin 43 is involved in aldosterone-induced podocyte injury.
Connexin43 (Cx43) belongs to a family of transmembrane proteins that build cell-to-cell channels in gap junctions, which play an essential role in intercellular communication, cell proliferation, differentiation and apoptosis. The aim of this study is to clarify the effect of Cx43 in aldosterone (Aldo)-induced podocyte injury and explore the possible molecular mechanism behind this effect. Uninephrectomized Sprague-Dawley rats were given 1% NaCl (salt) in their water and an Aldo infusion (0.75 μg/h) for 28 days to induce podocyte injury. Podocytes were incubated in media containing either buffer or increasing concentrations of Aldo (10(-9) - 10(-7) M) for variable time periods. The podocytes were then examined and the mechanism of injury investigated using TUNEL assay, ELISA, immunofluorescence staining, western blot, RNA interference, and DCFDA fluorescence. Here, we report that in vivo administration of Aldo caused greater numbers of TUNEL-positive podocytes, accompanied by increased Cx43 expression, but a reduction in WT1-positive podocytes. In vitro studies indicated that Aldo induces podocyte apoptosis in a dose- and time-dependent manner and is accompanied by increases in Cx43 expression, gap junction intercellular communication (GJIC), ROS production, and the Bax/Bcl-2 ratio. Silencing of Cx43 expression attenuates the increase of ROS production and the subsequent up-regulation of the Bax/Bcl-2 ratio, which partially inhibited the podocytes apoptosis. Our study provides preliminary evidence that upregulation of Cx43 expression is involved in Aldo-induced podocyte injury, and that Cx43 is a potentially relevant therapeutic target for the treatment of chronic kidney diseases (CKD).


How Rap and its GEFs control liver physiology and cancer development. C3G alterations in human hepatocarcinoma.
Rap proteins regulate liver physiopathology. For example, Rap2B promotes hepatocarcinoma (HCC) growth, while Rap1 might play a dual role. The RapGEF, Epac1, activates Rap upon cAMP binding, regulating metabolism, survival, and liver regeneration. A liver specific Epac2 isoform lacking cAMP-binding domain also activates Rap1, promoting fibrosis in alcoholic liver disease. C3G (RapGEF1) is also present in the liver, but mainly as shorter isoforms. Its function in the liver remains unknown. Information from different public genetic databases revealed that C3G mRNA levels increase in HCC, although they decrease in metastatic stages. In addition, several mutations in RapGEF1 gene are present, associated with a reduced patient survival. Based on this, C3G might represent a new HCC diagnostic and prognostic marker, and a therapeutic target.


The Suppression Induced S-NDR Mechanism for Defect-Free Filling of High Aspect Ratio Features.
Superconformal electrodeposition in Through Silicon Vias for multiple metal systems is summarized and explained using models based on suppressor-induced, S-shaped negative differential resistance (S-NDR). Recent results for Ni are detailed, and the filling morphology is compared to previously published observations of superconformal Au, Cu and Zn in TSVs. Voltammetric measurements for quantifying the kinetics and interactions of metal deposition and suppressor adsorption are shown. Dependence of the superconformal filling behavior on deposition conditions including suppressor concentration, transport and deposition potential are described. S-NDR models based on fractional coverage of adsorbed suppressor capture experimental trends and predict the filling geometries. The results demonstrate the generality of the S-NDR mechanism for achieving void-free filling of large features by electrodeposition and the power of S-NDR based models for prediction of filling evolution and process design.


Emergency management of traumatic dental injuries in 42 countries.
Availability and quality of emergency services for traumatic dental injuries (TDI) are important as the prognosis of TDI is decided at the place of accident or during the first hours after the injury has occurred. To report how emergency services for TDI are provided in some places around the world. This is a preliminary observational study of descriptive character. One-hundred and three participants from 83 cities in 42 countries described the emergency services in their city by answering a questionnaire related to: (1) availability of advice per telephone at the place of the accident; (2) availability of emergency treatment resources at different times of the day; and; (3) assessment of competence level of the person carrying out the service. Answers and comments were scored with regard to availability and level of competence. 62% of the participants were not satisfied with their current emergency services. 50% reported organized emergency telephone service always available. 43% of the advice given by phone was from professionals with dental training. An organized on-call 24/7 service was available in 33%. Treatment of TDI within the first hour was available in 53%. A competent dentist was available in 40%. 56% reported compromised competency outside office hours. Places with a well-organized system built on a well-developed first-line care managed by trauma-trained general dentists, ideally on a 24/7 basis, supported by on-call specialists outside office working hours, achieved the highest scores. With all the limitations of a preliminary questionnaire study, we can conclude that there is a need to perform more comprehensive well-designed studies in this field to understand more of the varying quality of emergency services worldwide. The major problem today seems to be related to limited availability and competence, especially outside office working hours.


Photoacoustic cell using elliptical acoustic focusing.
A photoacoustic cell has been developed in the form of an elliptical cylinder in which essentially all the acoustic energy generated by a laser beam passing down one axis is focused onto a cylindrical acoustic transducer located along the other axis. Preliminary measurements on a liquid-filled cell of this design show high sensitivity and a notably clean impulse response. A similar design may be useful for photoacoustic measurements in vapors as well.


Laying patterns of geese in the mid west.
In the nine years from 1966 through 1974 two large flocks of geese, one Emden and the other Toulouse, averaging respectively 1959 and 734 females began laying between January 9 and February 28 with the Toulouse lagging an average of 16 days. It took a mean of 17 days to advance from the 5% to the 33-1/3% level of laying. An average rate of 29.5% was thereafter maintained for 80 days, after which it dropped to 16% in ten more days.


Large Cretaceous sphenodontian from Patagonia provides insight into lepidosaur evolution in Gondwana.
Sphenodontian reptiles successfully radiated during Triassic and Jurassic times, but were driven almost to extinction during the Cretaceous period. The sparse Early Cretaceous record of sphenodontians has been interpreted as reflecting the decline of the group in favour of lizards, their suspected ecological successors. However, recent discoveries in Late Cretaceous beds in Patagonia partially modify this interpretation. Numerous skeletons of a new sphenodontian, Priosphenodon avelasi gen. et sp. nov., were collected from a single locality in the Cenomanian-Turonian Candeleros Formation, where it is more abundant than any other tetrapod group recorded in the quarry (for example, Crocodyliformes, Serpentes, Dinosauria and Mammalia). Adult specimens of Priosphenodon reached one metre in length, larger than any previously known terrestrial sphenodontian. Here we propose, using available evidence, that sphenodontians were not a minor component of the Cretaceous terrestrial ecosystems of South America, and that their ecological replacement by squamates was delayed until the early Tertiary. The new discovery helps to bridge the considerable gap in the fossil record (around 120 million years) that separates the Early Cretaceous sphenodontians from their living relatives (Sphenodon).


Trypsin-catalyzed Oligomerization of L-Lysine Esters.
The oligomerization of L-lysine esters with a free α-amino group was done using trypsin as a catalyst in aqueous media, and some reaction parameters were examined. The pH-dependence of the reaction suggested that aminolysis by the substrate species having non-protonated α- and ɛ-amino groups was a dominant factor governing the reaction progress. At the beginning of the reaction lysine oligomers containing up to 8 residues were observed, but much of the dimer was accumulated during prolonged incubation due to secondary hydrolysis of highly polymerized products. The reaction yield depended on the ratio of enzyme and substrate; an overall reaction yield higher than 70% was attained after 2 hr of reaction when 1,000 mM of L-lysine ethyl ester was treated with 200 μM of trypsin at pH 10.0. The oligomerization was apparently enhanced by an addition of NaCl and by using the longer alkyl esters.


[The European Society of Cardiology (ESC) guidelines for percutaneous coronary interventions (PCI). Three case reports].
The European Society of Cardiology (ESC) for the first time issued guidelines for percutaneous coronary interventions (PCI) in spring 2005. The strengths of recommendations stated in the ESC guidelines (as in those of the AHA/ACC [American Heart Association/American College of Cardiology]) are traditionally a combination of recommendation classes (I, IIa, and IIb) and a level of evidence (A, B, or C). This paper explains and discusses selected focal points of the ESC PCI guidelines based on three representative cases from daily practice. 1. Stable coronary artery disease (CAD): PCI in a 53-year-old patient without angina pectoris and proof of myocardial ischemia. With a clear indication of ischemia in the anterior myocardial wall, the ESC PCI guidelines indicated coronary angiography with possible PCI, even without angina pectoris symptoms. Cardiac catheterization showed a 99% proximal LAD stenosis, which was immediately dilated and stented based on the indicated ischemia. According to the ESC PCI guidelines, an intervention is indicated for CAD when a larger ischemic area is clearly evident even in the absence of typical angina (recommendation class I A). 2. ST segment elevation myocardial infarction (STEMI): PCI even after successful thrombolysis. A 70-year-old patient experienced acute substernal pain and immediately went to his nearby hospital. The ECG clearly showed anterior myocardial wall STEMI, which in this hospital without a cardiac cath lab indicated thrombolysis, since it could be initiated within 3 h after the onset of chest pain. Pain relief was evident soon after thrombolysis, combined with a resolution of the ST segment elevations. As suggested by the ESC PCI guidelines, a transfer to a cardiac cath lab took place the next day, where the 50% residual stenosis of the LAD was stented. The ESC PCI guidelines suggest coronary angiography with possible PCI within 1-2 days following successful thrombolysis (recommendation class I A). Thus, even "successful" thrombolysis is not regarded as the final treatment for STEMI. 3. Premature termination of clopidogrel after stent implantation: stent thrombosis with acute myocardial infarction. A 46-year-old patient visited the practice due to increasing dyspnea. 4 months earlier, a Taxus stent had been implanted at a heart center into the second RPLS of the RCX; 3 days later, a Cypher stent was implanted in the LAD. Upon being discharged on a Friday at noon, the patient was advised to see his general practitioner soon to attain a prescription for clopidogrel. The patient was given an appointment at his general practitioner for the following Wednesday afternoon. But on that Wednesday morning the patient went into cardiogenic shock. Although the occluded LAD (stent thrombosis) could be quickly reopened, left ventricular myocardium became severely damaged. Until a cardiac transplantation will be performed, a defibrillator was implanted. This "organizational" gap in clopidogrel administration did not conform to the ESC PCI guidelines: after implantation of any coronary stent, dual antiplatelet treatment (acetylsalicylic acid and clopidogrel) must be consistently administered for at least 4 weeks. After implantation of drug-eluting stents (DES), the ESC PCI guidelines call for clopidogrel administration for at least 6 months; when small vessels, long lesions or a complex anatomy (e. g., bifurcation stenting) are involved, a duration of 1 year or even longer is recommended. The optimal duration of platelet aggregation inhibition following PCI with DES of unprotected left main stem stenoses is unknown at this time. The traditional levels of evidence according to ESC, AHA and ACC criteria (levels A, B, or C) do no longer meet the actual requirements to assess the scientific evidence of randomized PCI trials and registry studies. For example, only two small randomized studies with few patients and insufficient statistical power utilizing a clinically insignificant surrogate endpoint would be enough to attain level of evidence A. Consequently, a new scoring system will be proposed, which considers criteria such as the importance of a primary clinical endpoint, the statistical power achieved, and the presence of an independent external data review and safety monitoring board.


Deciphering the Rules for Amino Acid Co-Assembly Based on Interlayer Distances.
Metabolite materials are extremely useful to obtain functional bioinspired assemblies with unique physical properties for various applications in the fields of material science, engineering, and medicine by self-assembly of the simplest biological building blocks. Supramolecular co-assembly has recently emerged as a promising extended approach to further expand the conformational space of metabolite assemblies in terms of structural and functional complexity. Yet, the design of synergistically co-assembled amino acids to produce tailor-made functional architectures is still challenging. Herein, we propose a design rule to predict the supramolecular co-assembly of naturally occurring amino acids based on their interlayer separation distances observed in single crystals. Using diverse experimental techniques, we demonstrate that amino acids with comparable interlayer separation strongly interact and co-assemble to produce structural composites distinctly different from their individual properties. However, such an interaction is hampered in a mixture of differentially layer-separated amino acids, which self-sort to generate individual characteristic structures. This study provides a different paradigm for the modular design of supramolecular assemblies based on amino acids with predictable properties.


Time to awakening after cardiac arrest and the association with target temperature management.
Target temperature management (TTM) at 32-36 °C is recommended in unconscious survivors of cardiac arrest. This study reports awakening in the TTM-trial. Our predefined hypotheses were that time until awakening correlates with long-term neurological outcome and is not affected by level of TTM. Post-hoc analysis of time until awakening after cardiac arrest, its association with long-term (180-days) neurological outcome and predictors of late awakening (day 5 or later). The trial randomized 939 comatose survivors to TTM at 33 °C or 36 °C with strict criteria for withdrawal of life-sustaining therapies. Administered sedation in the treatment groups was compared. Awakening was defined as a Glasgow Coma Scale motor score 6. 496 patients had registered day of awakening in the ICU, another 43 awoke after ICU discharge. Good neurological outcome was more common in early (275/308, 89%) vs late awakening (142/188, 76%), p < 0.001. Awakening occurred later in TTM33 than in TTM36 (p = 0.002) with no difference in neurological outcome, or cumulative doses of sedative drugs at 12, 24 or 48 h. TTM33 (p = 0.006), clinical seizures (p = 0.004), and lower GCS-M on admission (p = 0.03) were independent predictors of late awakening. Late awakening is common and often has a good neurological outcome. Time to awakening was longer in TTM33 than in TTM36, this difference could not be attributed to differences in sedative drugs administered during the first 48 h.


Dose-response hyperplasia and neoplasia from feeding N-2-fluorenylacetamide (2-FAA) to BALB/c mice for varying time intervals.
Hyperplasia induced by N-2-fluorenylacetamide requires the continuous presence of the chemical for its maintenance. Feeding the chemical at levels of 8 to 86 ppm results in a dose-response related urinary bladder hyperplasia and, after 9 to 12 months, in bladder neoplasia at the higher dose levels. Neoplasia of the lungs, liver, parotid gland, lacrimal gland, harderian gland, and submaxillary gland was also observed. Measurement of oncogenic viruses revealed no synergism with 2-FAA and no relationship between virus counts and neoplasia. Calcareous pericarditis was observed and most general toxicological effects occurred in the liver. A pronounced sex difference in response was observed with the males giving greater responses than the females in all instances.


Protein crystals and charged surfaces: interactions and heterogeneous nucleation.
As proteins typically have charges of around 10, they will interact strongly with charged surfaces. We calculate the electrostatic contribution to the interaction of crystals of protein with charged surfaces. The surfaces repel like-charged crystals and attract oppositely charged crystals, with free energies that can be easily several kT per protein molecule brought into contact with the surface. This means that oppositely charged surfaces can act as a nucleant, they can induce nucleation of a protein crystal by lowering the free energy barrier to heterogeneous nucleation of the crystal from a dilute solution.


Accuracy of LightCycler(®) SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review and meta-analysis.
There is an urgent need to develop diagnostic tests to improve the detection of pathogens causing life-threatening infection (sepsis). SeptiFast is a CE-marked multi-pathogen real-time PCR system capable of detecting DNA sequences of bacteria and fungi present in blood samples within a few hours. We report here a systematic review and meta-analysis of diagnostic accuracy studies of SeptiFast in the setting of suspected sepsis. A comprehensive search strategy was developed to identify studies that compared SeptiFast with blood culture in suspected sepsis. Methodological quality was assessed using QUADAS. Heterogeneity of studies was investigated using a coupled forest plot of sensitivity and specificity and a scatter plot in receiver operator characteristic space. Bivariate model method was used to estimate summary sensitivity and specificity. From 41 phase III diagnostic accuracy studies, summary sensitivity and specificity for SeptiFast compared with blood culture were 0.68 (95 % CI 0.63-0.73) and 0.86 (95 % CI 0.84-0.89) respectively. Study quality was judged to be variable with important deficiencies overall in design and reporting that could impact on derived diagnostic accuracy metrics. SeptiFast appears to have higher specificity than sensitivity, but deficiencies in study quality are likely to render this body of work unreliable. Based on the evidence presented here, it remains difficult to make firm recommendations about the likely clinical utility of SeptiFast in the setting of suspected sepsis.


Polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine (Pinus virginiana Mill.) plantlets.
Polyamines have been demonstrated to play an important role in adventitious root formation and development in plants. Here, we present a detailed analysis of influence of exogenously added polyamines on adventitious root development and its relationship to cold tolerance in Virginia pine (Pinus virginia Mill.). Our results demonstrated that polyamines putrescine (Put), spermidine (Spd), and spermine (Spm) at 0.001 mM improve rooting frequency and promote root elongation. Put, Spd, and Spm at 0.01-1 mM decrease rooting frequency and reduce root elongation root elongation. Measurements of diamine oxidase (DAO, EC 18.104.22.168) and polyamine oxidase (PAO, EC 18.104.22.168) activities showed that higher DAO and PAO enzyme activities were obtained when high concentrations of polyamines were applied and when plantlets were treated for 5-7 week at 4 degrees C and 16 degrees C. Survival rate of plantlets increased with the treatment of polyamines at low temperature. Polyamines increased mitotic index of cells in root tips of regenerated plantlet cultured on medium containing 0.001 microM Put, Spd, or Spm, but did not increase mitotic index in tissues of needle tips of the same plantlets. These results demonstrated that polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine plantlets.


Iridoschisis: a case report and literature review.
Iridoschisis is a rare condition that is characterized by the separation of the iris stroma into layers, with portions of the anterior layer floating freely in the aqueous humour. Here, we report three cases of iridoschisis that were complicated by either a cataract or glaucoma. Based on these cases, we speculate that the scope of iridoschisis has a rare association with intraocular pressure and the loss of corneal endothelial cells after surgery, which is mainly due to the surgery and not iridocorneal mechanical contact. We report three cases of iridoschisis, two of which were complicated by cataracts and the third by glaucoma. Patient 1 was a 69-year-old man with bilateral iridoschisis complicated by a cataract but not glaucoma, even though the entire anterior layer of the iris stroma in the right eye presented as white atrophic strands. To prevent the detached iris fibrils from invading the phacoemulsification tip and the irrigation/aspiration hand piece port, the separated iris stroma that was floating freely in the aqueous humour was cut with scissors immediately before the cataract extraction. Patient 2 was an 87-year-old woman with iridoschisis complicated by a cataract in the right eye. We successfully performed cataract surgery on the right eye without a pupillary device. Patient 3 was a 66-year-old man who presented with increased intraocular pressure with bilateral iridoschisis. He was discharged and prescribed with a combination of four glaucoma drugs. Patients with iridoschisis should have continuous follow-up because complications may occur, and extra care from ophthalmologists is required.


Preparation of fullerene (C60) based SiO2 sonogel hybrid composites: UV laser induced photo-polymerization, morphological, and optical properties.
Fullerene (C60) spheres were successfully embedded by diffusion into a catalyst-free SiO2 sonogel network. By this method, homogeneous and stable solid state hybrid samples suitable for optical characterization were produced. Due to the high porosity exhibited by the sonogel matrix on the nanometric length-scale, the preparation of several C60 doped composites with variable dopant concentrations was achieved. The obtained bulk hybrids were irradiated with a pulsed Kr-F laser system in order to induce photo-polymerization of the implanted C60 guest molecules at optimal experimental conditions established according to the pulsed laser photoacoustic technique (LPAT). The adequate purity level displayed by the sonogel route allowed us to perform optimal optical characterizations of these composites (reference hybrids and photo-polymerized samples) in order to evaluate their photo-physical properties for potential photonic applications. In this work the sonolysis process used to prepare amorphous catalyst-free SiO2:C60 sonogel hybrids and their linear and nonlinear optical (NLO) properties are extensively discussed. UV-VIS absorption-, Photo Luminescent (PL)-, and Raman-spectroscopies in conjunction with nonlinear optical limiting (OL) measurements, gave experimental evidence of the inclusion of C60 molecules within the sonogel environment, the laser induced photo-polymerization process and the induced changes of the optical and NLO-properties. Results are discussed in combination with atomic force microscopy (AFM) studies in order to explore the surface morphology of these samples.


Regadenoson-induced T-wave heterogeneity complements coronary stenosis detection by myocardial perfusion imaging in men and women.
We analysed whether incorporating electrocardiographic interlead T-wave heterogeneity (TWH) with myocardial perfusion imaging (MPI) during pharmacologic stress improves detection of flow-limiting lesions (FLL). Medical records of all 103 patients at our institution who underwent stress testing with regadenoson (0.4 mg IV bolus) within 3 months of coronary angiography from September 2017 to March 2019 were studied. Cases (N = 59) had angiographically significant FLL (≥50% of left main or ≥70% of other epicardial coronary arteries ≥2 mm in diameter); controls (N = 44) were normal or had non-FLL. TWH, i.e., interlead splay of T waves, was assessed from precordial leads V4-6 by second central moment analysis. Maximum TWHV4-6 levels during regadenoson stress were 68% higher in cases than in controls (P < 0.0001). TWHV4-6 generated areas under the receiver-operating characteristic (ROC) curve of 0.79 in men (P < 0.0001) and 0.71 in women (P = 0.007). In men, the ROC-guided 54-µV TWHV4-6 cut-point for FLL produced adjusted odds of 7.3 [95% confidence interval (CI): 1.3-41.5, P = 0.03], 79% sensitivity, and 78% specificity. In women, the ROC-guided 35-µV TWHV4-6 cut-point produced adjusted odds of 4.5 (95% CI: 1.1-18.9, P = 0.04), 84% sensitivity, and 52% specificity. Adding TWHV4-6 to MPI determinations reduced false-positive results by 70%, more than doubled true-negative results, and improved adjusted odds ratio to 6.8 (95% CI: 2.2-21.4, P = 0.001) with specificity of 78% in men and 86% in women. This observational study is the first to demonstrate the benefit of combining TWHV4-6 with MPI to enhance FLL detection during MPI with regadenoson in both men and women.


Rapid and Simultaneous Analysis of 360 Pesticides in Brown Rice, Spinach, Orange, and Potato Using Microbore GC-MS/MS.
A multiresidue method for the simultaneous and rapid analysis of 360 pesticides in representative agricultural produce (brown rice, orange, spinach, and potato) was developed using a modified QuEChERS procedure combined with gas chromatography-tandem mass spectrometry (GC-MS/MS). Selected reaction monitoring transition parameters (e.g., collision energy, precursor and product ions) in MS/MS were optimized to achieve the best selectivity and sensitivity for a wide range of GC-amenable pesticides. A short (20 m) microbore (0.18 mm i.d.) column resulted in better signal-to-noise ratio with reduced analysis time than a conventional narrowbore column (30 m × 0.25 mm i.d.). The priming injection dramatically increased peak areas by masking effect on a new GC liner. The limit of quantitation was <0.01 mg/kg, and the correlation coefficients (r2) of matrix-matched standards were >0.99 within the range of 0.0025-0.1 mg/kg. Acetonitrile with 0.1% formic acid without additional buffer salts was used for pesticide extraction, whereas only primary-secondary amine (PSA) was used for dispersive solid phase extraction (dSPE) cleanup, to achieve good recoveries for most of the target analytes. The recoveries ranged from 70 to 120% with relative standard deviations of ≤20% at 0.01 and 0.05 mg/kg spiking levels (n = 6) in all samples, indicating acceptable accuracy and precision of the method. Seventeen real samples from local markets were analyzed by using the optimized method, and 14 pesticides in 11 incurred samples were found at below the maximum residue limits.


Pattern of sensitization to common environmental allergens amongst atopic Singapore children in the first 3 years of life.
This study was aimed to investigate the sensitization pattern to a range of common allergens in young Singaporean children. A cross-sectional study involving 75 children aged below 3 years was carried out. They presented between December 1995 and April 2000 with symptoms of asthma, rhinitis, eczema, or food allergy. Their levels of allergen-specific serum IgE to a panel of foods (egg white, milk, soy protein, shrimp, wheat and peanut), pet dander, dust mites and cockroaches were measured with Pharmacia CAP System radioallergosorbent test kits. Serum IgE levels greater than 0.35 kU/l represented a positive result. Four children could not be tested with the complete panel because of insufficient serum. The prevalence of sensitization was highest for cow's milk (45.9%) followed by egg white (38.7%), dust mites Dermatophagoides pteronyssinus (31.4%) and Blomia tropicalis (25.5%). Sensitization to ingested allergens was significantly more prevalent in children aged 1 year or younger than in the older children (70.4% of those below 1 year, and 50% of those aged 1-3 years; p < 0.02). Sensitization to inhaled allergens, such as dust mites, was more likely to manifest as respiratory symptoms (allergic rhinitis and asthma), while ingested allergens were associated with gastrointestinal symptoms and eczema (p < 0.001). It was concluded that infants and young children are at high risk of sensitization to common environmental substances. Allergen avoidance is therefore important even in the very young. The prevalence of sensitization to food allergens is higher compared to inhalant allergens in young children.


Retinal pigment epithelium adenoma in vitreous fluid cytology.
Ocular cytology specimens are relatively uncommon, adding to the difficulty of their evaluation by cytopathologists. While melanomas account for a majority of primary intraocular pigmented lesions, other diagnostic considerations must be included in the differential. This brief report highlights a case of a pigmented ocular lesion in a 24-year-old man and key morphologic, immunohistochemical, and clinical differences between melanoma, melanocytoma, choroidal nevus, and retinal pigment epithelium (RPE) adenoma.


[PRELIMINARY EFFECTIVENESS OF POLYAMINOACID/NANO-HYDROXYAPATITE/CALCIUM SULFATE CAGE IN LUMBAR FUSION SURGERY].
To discuss the early effectiveness of polyaminoacid/nano-hydroxyapatite/calcium sulfate (PAA/HA/CS) Cage (PHC Cage) in lumbar fusion surgery. Thirty cases undergoing lumbar fusion of single segment between March and September 2014 were enrolled in this study. The patients were randomly divided into the trial group (n = 20) and the control group (n = 10). The PHC Cage was implanted in the trial group, while the polyetheretherketone (PEEK) Cage was implanted in the control group. The patients of 2 groups mainly presented lumbocrural pain and lower limb radiation pain or numbness. There was no significant difference in gender, age, type, affected segment, disease duration, preoperative intervertebral height, the lordosis angle of fusion segments, and the Oswestry Disability Index (ODI) between 2 groups (P > 0.05). Lateral lumbar X-ray films and three dimensional CT were taken preoperatively and at 1 week and 3, 6, and 12 months postoperatively. The intervertebral height and the lordosis angle of fusion segments at 1 week and 3, 6, and 12 months after operation and ODI at 3, 6, and 12 months after operation were measured; and the bone graft fusion rate was evaluated according to Brantigan criteria. There was no significant difference in operation time, intraoperative blood loss, and the amount of autologous blood transfusion between 2 groups (P > 0.05). Healing by first intention as obtained in 30 cses. All patients were followd up 12 months. The intervertebral height of fusion segments, the lordosis angle of fusion segments, and ODI at each time point after operation were significantly improved when compared with preoperative ones (P < 0.05). The ODI showed significant difference between 3 months and 6, 12 months (P < 0.05), but there was no significant difference between the other time points after operation (P > 0.05). There was no significant difference in the intervertebral height and the lordosis angle of fusion segments between groups at different time points (P > 0.05). There was no significant difference in the above indexes between the trial group and the control group at each time point (P > 0.05). At last follow-up, 5 cases were rated as Brantigan grade E, 13 cases as grade D, and 2 cases as grade C in the trial group; 4 cases were rated grade E, 5 cases as grade D, and 1 case as grade C in the control group. The bone fusion rate was 90% in 2 groups. The PHC Cage can effectively restore and maintain the disc height of fusion segment, normal sequence and biomechanical stability of the lumbar spine. The PHC Cage is similar to the PEEK Cage and has good clinical outcome in short-term follow-up.


To clip or to coil for unruptured intracranial aneurysm?: A protocol of randomized controlled trial.
Microsurgical clipping and endovascular coiling are the main methods against unruptured intracranial aneurysm (UIA). The craniotomy of surgical clipping may increase the risk of cerebrospinal fluid leakage and infection, damage the brain tissue, produce excessive stimulation to the nerves and blood vessels around the aneurysm, and cause the corresponding neurological deficit. Endovascular coiling could significantly reduce the mortality and disability rate than surgical clipping technique, which made endovascular coiling to become the first choice for the treatment of UIA. However, the long-term results showed attenuated favorable outcomes of coiling over clipping, so it is still in debate whether to clip or to coil. Therefore, we try to conduct a randomized, controlled, prospective trial to assess the long term safety of endovascular coiling therapy against UIA compared with microsurgical clipping technique. Parallel-group randomization (1:1) is generated through the random number generator in Microsoft Excel 2010. In this trial, blinding to patients, physicians, and outcome assessors is not possible. Endovascular coiling or surgical clipping will be performed once for each patient in treatment group or control group, respectively. The mRS, overall mortality rate, disability rate, morbidity rate, and occurrence of a major aneurysm recurrence measured at 6 month and 1 year will be recorded. The findings will be helpful for the choice of endovascular coiling or surgical clipping by assessing the long term efficacy and safety of both operations against UIA. OSF Registration number: DOI 10.17605/OSF.IO/QYE9F.


[Footballer's migraine instead of concussion].
Our patient, a 27-year-old amateur soccer player, was hit hard against the head by a ball. Within a few minutes he developed visual and sensory symptoms followed by headache, nausea, vomiting, photophobia and phonophobia. After treatment with paracetamol and metoclopramide, he was free of symptoms within 24 hours. This picture is stereotypically associated with migraine provoked by minor head injury. TTM may also occur without aura. Trauma triggered migraine is seen more frequently in children, adolescents and young adults. The cause of trauma triggered migraine is unknown. Treatment of the headache and nausea with common analgesics and anti-emetic drugs might be effective. Proper explanation and reassurance are most important.


A dual-phase alloy with ultrahigh strength-ductility synergy over a wide temperature range.
High-entropy alloys (HEAs), as an emerging class of materials, have pointed a pathway in developing alloys with interesting property combinations. Although they are not exempted from the strength-ductility trade-off, they present a standing chance in overcoming this challenge. Here, we report results for a precipitation-strengthening strategy, by tuning composition to design a CoNiV-based face-centered cubic/B2 duplex HEA. This alloy sustains ultrahigh gigapascal-level tensile yield strengths and excellent ductility from cryogenic to elevated temperatures. The highest specific yield strength (~150.2 MPa·cm3/g) among reported ductile HEAs is obtained. The ability of the alloy presented here to sustain this excellent strength-ductility synergy over a wide temperature range is aided by multiple deformation mechanisms i.e., twins, stacking faults, dynamic strain aging, and dynamic recrystallization. Our results open the avenue for designing precipitation-strengthened lightweight HEAs with advanced strength-ductility combinations over a wide service temperature range.


Evaluation of canonical Hedgehog signaling pathway inhibition in canine osteosarcoma.
Canine osteosarcoma (OSA), the most common canine primary bone malignancy, has a highly aggressive biologic behavior. Despite current standard of care therapies, including amputation and adjuvant chemotherapy, most dogs still succumb to metastatic disease. Further investigations into molecular mechanisms and pathways driving OSA are needed to improve therapeutic options. The Hedgehog (HH) cell-signaling pathway has demonstrated involvement in human OSA. Several studies in canine OSA have found changes in expression of some HH pathway genes and demonstrated a role for HH transcription factors. However, the role of this pathway as well as the translational value of its targeting in canine OSA are still undefined. The objectives of this study were to determine the expression of HH components directly in canine OSA tissues and to evaluate the biologic impact of HH signaling inhibition in canine OSA cells. In situ hybridization was used to detect HH family mRNA expression in archived canine OSA tissues and revealed variable expression levels of these mRNAs in canine OSA tissues. The effect of a commercially available Smoothened inhibitor, vismodegib, was studied in established canine OSA cell lines. Alterations in cellular growth as well as assessment of downstream HH targets were evaluated. Although changes in cell growth were noted following Smoothened inhibition, inconsistent decreases in target gene expression were found. While treatment with vismodegib had a negative impact on canine OSA cell growth and viability, the mechanism remains unclear. Further studies are warranted to evaluate the clinical significance of canonical HH signaling in canine OSA.


Metagenome complexity and template length are the main causes of bias in PCR-based bacteria community analysis.
Multitemplate PCR is used widely for the study of microbial community diversity. Although such studies have established the abundance of different groups within many natural ecosystems, these reports are limited by uncertainties such as bias and artifacts in the PCR. Bias which is introduced by the simultaneous amplification of specific genes from complex mixtures of templates remains poorly understood. In this study, factors leading to the bias of the multitemplate PCR in bacterial communities were examined and optimized. Comparisons between PCR cycle parameters, DNA polymerases, PCR primer degeneracy, and 16S rRNA gene fragments GC content, revealed that annealing temperatures and DNA structure are predominant factors contributing to the observed bias. Pre-digestion of metagenomic DNA with the restriction enzyme Sau3A I and decreased annealing temperature reduced the bias significantly. The application of these optimized conditions to the ten-species model community in a soil sample verified the validity of these treatments.


Effects of cold cardioplegia on pH, Na, and Ca in newborn rabbit hearts.
Many studies suggest myocardial ischemia-reperfusion (I/R) injury results largely from cytosolic proton (H(i))-stimulated increases in cytosolic Na (Na(i)), which cause Na/Ca exchange-mediated increases in cytosolic Ca concentration ([Ca]i). Because cold, crystalloid cardioplegia (CCC) limits [H]i, we tested the hypothesis that in newborn hearts, CCC diminishes H(i), Na(i), and Ca(i) accumulation during I/R to limit injury. NMR measured intracellular pH (pH(i)), Na(i), [Ca]i, and ATP in isolated Langendorff-perfused newborn rabbit hearts. The control ischemia protocol was 30 min for baseline perfusion, 40 min for global ischemia, and 40 min for reperfusion, all at 37 degrees C. CCC protocols were the same, except that ice-cold CCC was infused for 5 min before ischemia and heart temperature was lowered to 12 degrees C during ischemia. Normal potassium CCC solution (NKCCC) was identical to the control perfusate, except for temperature; the high potassium (HKCCC) was identical to NKCCC, except that an additional 11 mmol/l KCl was substituted isosmotically for NaCl. NKCCC and HKCCC were not significantly different for any measurement. The following were different (P < 0.05). End-ischemia pH(i) was higher in the CCC than in the control group. Similarly, CCC limited increases in Na(i) during I/R. End-ischemia Na(i) values (in meq/kg dry wt) were 115 +/- 16 in the control group, 49 +/- 13 in the NKCCC group, and 37 +/- 12 in the HKCCC group. CCC also improved [Ca]i recovery during reperfusion. After 40 min of reperfusion, [Ca](i) values (in nmol/l) were 302 +/- 50 in the control group, 145 +/- 13 in the NKCCC group, and 182 +/- 19 in the HKCCC group. CCC limited ATP depletion during ischemia and improved recovery of ATP and left ventricular developed pressure and decreased creatine kinase release during reperfusion. Surprisingly, CCC did not significantly limit [Ca]i during ischemia. The latter is explained as the result of Ca release from intracellular buffers on cooling.


Hemilaminectomy for removal of extramedullary or extradural spinal cord tumors: medium to long-term clinical outcomes.
Laminectomy is generally the treatment of choice for removal of spinal tumors. However, it has been shown that laminectomy may cause instability due to damage of posterior elements of the spinal column, which may induce subsequent kyphosis in the future. Therefore, to reduce the risk of deformity and spinal instability after laminectomy, hemilaminectomy has been used. However, the medium to long-term effects of hemilaminectomy on spinal sagittal alignment is not well understood. The present study was performed to evaluate the clinical outcomes, including spinal sagittal alignment of patients, associated with spinal cord tumors treated by surgical excision using hemilaminectomy. Twenty hemilaminectomy operations at our institute for extramedullary or extradural spinal cord tumors in 19 patients were evaluated retrospectively with an average follow-up of 85 months (range, 40-131 months). Neurological condition was evaluated using the improvement ratio of the Japanese Orthopaedic Association Score (JOA score) for cervical, thoracic myelopathy, or back pain, and sagittal alignment by sagittal Cobb angle of the hemilaminectomied area. The mean improvement ratio of neurological results was 56.7% in the cervical spine (p < 0.01, n = 10), 26.3% in the thoracic spine (not significant, n = 5), and 48.6% in the lumbar spine (NS, n = 5). The sagittal Cobb angle was 4.3 ± 18.0° in the preoperative period and 5.4 ± 17.6° at the latest follow-up, indicating no significant deterioration. Hemilaminectomy is useful for extramedullary or extradural spinal cord tumors in providing fair neurological status and restoration of spinal sagittal alignment in medium to long-term follow-up.


Removal of cranial springs after spring-mediated cranioplasty.
Cranial spring hardware is generally removed 3 months after placement for spring-mediated cranioplasty. Spring removal is performed as an outpatient procedure under general anesthesia in approximately 15 minutes through the incision locations of the index procedure. Herein, the authors provide a multimedia demonstration of cranial spring hardware removal after spring-mediated cranioplasty for sagittal craniosynostosis. The video can be found here: https://vimeo.com/511179695.


Bacteremia in a general hospital. A prospective study of 102 consecutive cases.
A prospective clinical-bacteriological study of 102 consecutive cases of confirmed bacteremia at a Copenhagen City general hospital was carried out during 5 months of 1973 with special concern given to focus of infection and acquisition of microorganisms. Valid positive cultures were obtained from 7.2 patients per 1000 admissions. 50 of the 102 bacteremias were by all probability acquired in the hospital, mainly due to transurethral manipulations or intravenous lines. Pneumonia and hepatobiliary infections accounted for most of the non-hospital acquired bacteremias. 26/102 patients died in relation to the bacteremia. Escherichia coli and Staphylococcus aureus caused more than half of the infections. Bacteremia caused by proteus, klebsiella, enterobacter species of staphylococci was in most cases nosocomial and carried the highest mortality, i.e. 40%, verus 15% when other organisms were responsible. It is concluded that nosocomial bacteremia is a frequent and life-endangering complication which is often preceded by certain diagnostic or therapeutic procedures, not invariably linked to severe underlying diseases. Consequently, attempts to reduce bacteremic episodes should include surveillance of ecological factors and certain hospital procedures.


[Slide tracheoplasty in congenital tracheal stenosis].
Slide Tracheoplasty has progressively become the gold standard in the management of long-segment tracheal stenosis in children and infants. However, in certain situations this operation might become difficult and others techniques might be preferred. Five patients, 1 to 6 month old had surgery by our team, for complex tracheal stenosis between 2001 and 2005. Tracheal hypoplasia was considered complex because it was associated to either: an acquired critical stenosis, a cricoid stenosis, a bronchial stenosis, a tracheal bronchus or associated to oesophageal atresia and severe tracheomalacia. All of them have been treated by a modified slide tracheoplasty. The postoperative status required an average of 15 days in ICU, including 8 days of ventilation. Two patients had laryngeal nerve injury. Medium term follow up (27 months) demonstrated no need for re-intervention, good potential growth and normal child activity. Slide tracheoplasty can provide good results in certain cases of severe and complex tracheal hypoplasia in infants.


Tissue-resident Eomes(hi) T-bet(lo) CD56(bright) NK cells with reduced proinflammatory potential are enriched in the adult human liver.
The adult human liver is enriched with natural killer (NK) cells, accounting for 30-50% of hepatic lymphocytes, which include tissue-resident hepatic NK-cell subpopulations, distinct from peripheral blood NK cells. In murine liver, a subset of liver-resident hepatic NK cells have altered expression of the two highly related T-box transcription factors, T-bet and eomesodermin (Eomes). Here, we investigate the heterogeneity of T-bet and Eomes expression in NK cells from healthy adult human liver with a view to identifying human liver-resident populations. Hepatic NK cells were isolated from donor liver perfusates and biopsies obtained during orthotopic liver transplantation (N = 28). Hepatic CD56(bright) NK cells were Eomes(hi) T-bet(lo) , a phenotype virtually absent from peripheral blood. These NK cells express the chemokine receptor CXCR6 (chemokine (C-X-C motif) receptor 6), a marker of tissue residency, which is absent from hepatic CD56(dim) and blood NK cells. Compared to blood populations, these hepatic CD56(bright) NK cells have increased expression of activatory receptors (NKp44, NKp46, and NKG2D). They show reduced ability to produce IFN-γ but enhanced degranulation in response to challenge with target cells. This functionally distinct population of hepatic NK cells constitutes 20-30% of the total hepatic lymphocyte repertoire and represents a tissue-resident immune cell population adapted to the tolerogenic liver microenvironment.


Unilateral tubular approach for bilateral laminotomy: effect on ipsilateral and contralateral buttock and leg pain.
Tubular laminotomy is an effective procedure for treatment of lumbar spinal stenosis (LSS) and lateral recesses stenosis. Most surgeons familiar with the procedure agree that the tubular approach appears to afford a more complete decompression of the contralateral thecal sac and nerve root, as compared to the ipsilateral approach. With this study we sought to answer the question whether this is reflected in clinically significant differences between the ipsilateral and contralateral side pain improvements. In a retrospective case study, patients with LSS and lateral recesses stenosis who started out with VAS scores that were similar on the right and left side were included. All patients underwent a tubular (MIS) "over the top" laminotomy from a unilateral approach and through one incision. Surgeries were performed by a single surgeon in a single center. At the last follow-up, the extent of VAS score improvement on the approach (ipsilateral) side was compared to that of the contralateral side. Thirty-three patients were included in. At the latest follow-up of 25.8 ± 3.4 months, there were statistically significant improvements in ODI and back VAS scores (p = 0.002 and p < 0.0001, respectively). In addition, buttock VAS scores were significantly improved both on the ipsilateral and the contralateral side (p < 0.001, and p = 0.001, respectively). Similarly, leg VAS scores were improved significantly on both sides (p < 0.001, and p = 0.001, respectively). There were no statistically significant differences between the extent of pain improvement on the ipsilateral and the contralateral side. MIS tubular laminotomy through a unilateral approach results in clinically effective bilateral decompression of LSS and lateral recesses, regardless of the approach side.


Differences between Elite and Semi-Elite Australian Football Conceptualised through the Lens of Ecological Dynamics.
This study explored the differences in match play between elite and semi-elite Australian football (AF) conceptualised through the lens of ecological dynamics. We sampled naturalistic constraints from match play across two AF competitions (elite and semi-elite) and heuristically classified them into task, environmental and individual classes. Data was extracted from 22 Australian Football League (AFL) games, and 18 semi-elite AF games, with a total of six constraints being sampled from each game. Match play within the AFL generated a greater percent of total disposals in general play within a processing time of 0-1s (d = 1.24 (0.64-1.80)), a greater opposition density surrounding the ball carrier (d = 0.82 (0.26-1.37)), and more disposals being performed while running (dynamic; d = 0.89 (0.33-1.45)). This data highlights differences with regards to the informational sources available to players across both competition standards to inform their movement choices. Specifically, a greater proportion of disposals within the AFL appear to be shaped by pronounced temporal and spatial constraints relative to a semi-elite competition. Coaches are encouraged to consider these results when developing representative training activities for both AFL and prospective AFL players.


Newborn screening for Tyrosinemia type 1 using succinylacetone - a systematic review of test accuracy.
Tyrosinemia type 1 is an autosomal recessive disorder of amino acid metabolism. Without treatment, death in childhood is common. Treatment with nitisinone and dietary restrictions are associated with improved outcomes; some studies suggest better outcomes when treatment begins at an asymptomatic stage. Newborn screening allows for earlier identification, but there is uncertainty regarding the test accuracy of the current method: succinylacetone measurement in dried blood spots using tandem mass spectrometry. We conducted a systematic review of literature published up to January 2016. Two reviewers independently assessed titles, abstracts, full texts, and conducted quality appraisals. A single reviewer extracted data, which was checked by a second reviewer. Ten studies provided test accuracy data: five studies reporting screening experiences and five case-control studies. Sensitivity (29 cases in total) and specificity (34,403 controls in total) were 100% in the case-control studies, but could not be calculated in the studies reporting screening experiences due to a lack of follow-up of screen-negative babies. Positive predictive values in the screening experience studies ranged from 66.7% (2 true positive cases, 1 false positive case from ~500,000 people screened) to 100% (8 true positive cases from 856,671 people screened); negative predictive values could not be calculated. Positive and negative predictive values cannot be calculated from case-control studies. Screening for Tyrosinemia type 1 using tandem mass spectrometry measurement of succinylacetone from dried blood spots appears to be promising. Confirmation of test accuracy data should be obtained from studies that include a two-year follow-up of individuals who screen negative.


[Two Separate Instances of Anesthetic Management in a Patient Initially Diagnosed with Stiff-person Syndrome].
In this case report, we describe separate instances of general anesthetic management administered to a patient during treatment for two unrelated conditions. The patient, a 57-year-old woman who had been experiencing walking difficulties for about four years, fell down because of muscle rigidity and spasms and fractured her humerus. She was subsequently diagnosed with stiff-person syndrome. The fracture was treated conservatively, but three weeks later the alignment of the humerus became worse and the patient was scheduled to undergo an open reduction and internal fixation under general anesthesia (GA). Anesthesia was induced using propofol and fentanyl, and rocuronium was given for the tracheal intubation. The operation was completed successfully while anesthesia was maintained using sevoflurane and remifentanil without incident Four months later, the patient developed paraneoplastic syndrome stemming from breast cancer, and underwent a resection of the cancer under GA. The patient received propofol for the laryngeal mask insertion, and sevoflurane and pentazocine for maintenance of anesthesia. She was discharged from the hospital without any respiratory or airway problems. We used sevoflurane for maintenance of anesthesia on both occasions, and rocuronium for the first tracheal intubation. The patient's hemodynamics were stable during the operation. No prolonged effect of the muscle relaxant or any autonomic reactions were observed. The patient was fully conscious and in good respiratory condition after both surgeries, and was extubated in the operating room. There were no perioperative complications.


The role of syncytins in human reproduction and reproductive organ cancers.
Human life begins with sperm and oocyte fusion. After fertilization, various fusion events occur during human embryogenesis and morphogenesis. For example, the fusion of trophoblastic cells constitutes a key process for normal placental development. Fusion in the placenta is facilitated by syncytin 1 and syncytin 2. These syncytins arose from retroviral sequences that entered the primate genome 25 million and more than 40 million years ago respectively. About 8% of the human genome consists of similar human endogenous retroviral (HERVs) sequences. Many are inactive because of mutations or deletions. However, the role of the few that remain transcriptionally active has not been fully elucidated. Syncytin proteins maintain cell-cell fusogenic activity based on ENV: gene-mediated viral cell entry. In this review, we summarize how syncytins and their receptors are involved in fusion events during human reproduction. The significance of syncytins in tumorigenesis is also discussed.


Micro-RNAs 518d-3p and 618 Are Upregulated in Individuals With Type 1 Diabetes With Multiple Microvascular Complications.
Objective: To compare the serum micro-RNAs (miRNAs) profile of individuals with type 1 diabetes without microvascular complications vs. those with multiple severe microvascular complications, in order to identify epigenetically modulated pathways in these two groups of individuals. Research Design and Methods: A total of 10 subjects were selected among individuals followed in the Diabetes Outpatient Clinic and sorted according to the absence or presence of all microvascular complications. Samples from these participants were used for evaluation of serum miRNA expression profile employing a qRT-PCR assay with hydrolysis probes based on the Taqman Low Density Arrays (TLDA) system. The top six most differentially expressed miRNAs between the aforementioned groups were validated by qRT-PCR in additional 47 type 1 diabetes individuals sorted according to the absence or presence of all microvascular complications and matched for age, sex, degree of metabolic control, diabetes duration, and age at diagnosis. Results: Twenty one out of three hundred and seventy seven miRNAs were upregulated in the group of individuals with all microvascular complications vs. the group without complications. The following miRs were validated: 518-3p, 34a-5p, 126-5p, 425-5p, 618, and 139-5p and logistic regression analyses showed that miRNA-518-3p and miRNA-618 were positively associated with multiple microvascular complications after adjustment for age, sex, diabetes duration, HbA1c and use of statin, angiotensin-converting enzyme inhibitors and amlodipine. Conclusions: In this cohort of type 1 diabetes individuals, serum miR-518d-3p and miR-618 were upregulated in those with diabetes kidney disease, diabetes retinopathy, peripheral neuropathy, and cardiovascular autonomic neuropathy in comparison to individuals with no microvascular complications.


Absorption, distribution and excretion of radioactivity by male rats after a single intravenous or subcutaneous dose of 14C-azacitidine.
The absorption, distribution and excretion of radioactivity were investigated in male Sprague-Dawley rats after a single intravenous or subcutaneous dose of 14C-azacitidine. After subcutaneous administration, 14C-azacitidine was rapidly absorbed and the radioactivity was distributed to the tissues. The absorption of radioactivity after subcutaneous administration was 76.6% of that observed after intravenous administration. There were no marked differences in the tissue distribution of the radioactivity between administration routes. The concentrations of radioactivity in most tissues, including the spleen and bone marrow, which are sites of action of azacitidine, were higher than those in the plasma. Particularly high concentrations of radioactivity were detected in the spleen, kidney and liver. The accumulation of radioactivity in blood cells increased from 0.5 to 48 h. The binding of azacitidine to serum protein was low at <9%, and the cumulative urinary and fecal excretion of radioactivity for 168 h after intravenous or subcutaneous administration was >95% of the administered dose, indicating that radioactivity did not accumulate in the tissues. The radioactivity was mainly excreted in the urine.


High-Level Culturability of Epiphytic Bacteria and Frequency of Biosurfactant Producers on Leaves.
To better characterize the bacterial community members capable of biosurfactant production on leaves, we distinguished culturable biosurfactant-producing bacteria from nonproducers and used community sequencing to compare the composition of these distinct cultured populations with that from DNA directly recovered from leaves. Communities on spinach, romaine, and head lettuce leaves were compared with communities from adjacent samples of soil and irrigation source water. Soil communities were poorly described by culturing, with recovery of cultured representatives from only 21% of the prevalent operational taxonomic units (OTUs) (>0.2% reads) identified. The dominant biosurfactant producers cultured from soil included bacilli and pseudomonads. In contrast, the cultured communities from leaves are highly representative of the culture-independent communities, with over 85% of the prevalent OTUs recovered. The dominant taxa of surfactant producers from leaves were pseudomonads as well as members of the infrequently studied genus Chryseobacterium The proportions of bacteria cultured from head lettuce and romaine leaves that produce biosurfactants were directly correlated with the culture-independent proportion of pseudomonads in a given sample, whereas spinach harbored a wider diversity of biosurfactant producers. A subset of the culturable bacteria in irrigation water also became enriched on romaine leaves that were irrigated overhead. Although our study was designed to identify surfactant producers on plants, we also provide evidence that most bacteria in some habitats, such as agronomic plant surfaces, are culturable, and these communities can be readily investigated and described by more classical culturing methods. The importance of biosurfactant production to the bacteria that live on waxy leaf surfaces as well as their ability to be accurately assessed using culture-based methodologies was determined by interrogating epiphytic populations by both culture-dependent and culture-independent methods. Biosurfactant production was much more frequently observed in cultured communities on leaves than in other nearby habitats, such as soil and water, suggesting that this trait is important to life on a leaf by altering either the leaf itself or the interaction of bacteria with water. While pseudomonads were the most common biosurfactant producers isolated, this habitat also selects for taxa, such as Chryseobacterium, for which this trait was previously unrecognized. The finding that most epiphytic bacterial taxa were culturable validates strategies using more classical culturing methodologies for their study in this habitat.


Difficulties in the neuroscience of creativity: jazz improvisation and the scientific method.
Creativity is a fundamental and remarkable human capacity, yet the scientific study of creativity has been limited by the difficulty of reconciling the scientific method and creative processes. We outline several hurdles and considerations that should be addressed when studying the cognitive neuroscience of creativity and suggest that jazz improvisation may be one of the most useful experimental models for the study of spontaneous creativity. More broadly, we argue that studying creativity in a way that is both scientifically and ecologically valid requires collaboration between neuroscientists and artists.


In utero acetaminophen exposure and child neurodevelopmental outcomes: Systematic review and meta-analysis.
Acetaminophen is a frequently used analgesic for pain and fever. There have been reports of adverse neurodevelopmental outcomes associated with in utero acetaminophen exposure. However, it is unclear whether this association is related directly to acetaminophen use, or the reasons for use. To summarise the literature on the association between in utero acetaminophen exposure and child neurodevelopmental outcomes, and assess the extent to which the association is due to confounding by indication. OVID for Medline, Embase, and PsycINFO, and EBSCO for CINAHL, from inception to August 18, 2022. We searched for peer-reviewed, English-language studies on in utero acetaminophen exposure and child neurodevelopmental outcomes. Data were extracted using a standardised form created a priori, and quality was assessed using the Systematic Assessment of Quality in Observational Research. We generated pooled risk ratios (RR) for outcomes examined by ≥3 studies using random-effects models; outcomes that could not be meta-analysed were narratively summarised following Synthesis Without Meta-Analysis guidelines. Twenty-two studies including 23 cohorts were eligible (n = 367,775 total participants; median: 51.7% with acetaminophen exposure). Studies were primarily prospective cohort studies from Europe and the US, with attention deficit/hyperactivity disorder (ADHD) being the most common outcome. Quality assessments resulted in 13.6% of studies being classified as high, 59.1% as medium, 22.7% as low, and 4.5% as very low quality. In utero acetaminophen exposure was associated with an elevated risk of ADHD (unadjusted pooled RR 1.32, 95% confidence interval [CI] 1.20, 1.44; I2 = 47%, n = 7 studies), with little difference after adjusting for confounders, including indications for acetaminophen use (adjusted pooled RR 1.34, 95% CI 1.15, 1.55; I2 = 50%, n = 4 studies). Confounding by indication did not explain the association between in utero acetaminophen exposure and child ADHD. Further, high-quality research is needed on this and other neurodevelopmental outcomes.


Participation of histidine in biosynthesis of the pyrimidine moiety of thiamin in Saccharomyces cerevisiae.
The amide nitrogen atom of glutamine is incorporated into the pyrimidine moiety of thiamin in Escherichia coli and Saccharomyces cerevisiae. However, addition of casamino acids to the medium increases incorporation of the amide nitrogen atom of glutamine in E. coli, but decreases it in S. cerevisiae. This suggests that some amino acids other than glutamine in casamino acids are more direct precursors of the pyrimidine moiety in S. cerevisiae. To determine the direct precursor, we investigated the competitive effect of 14N-amino acids on the incorporation of 15NH4Cl into the pyrimidine moiety and found that histidine decreased the incorporation of 15N. Thus, histidine was concluded to be the direct precursor of the nitrogen atom of the pyrimidine moiety of thiamin in S. cerevisiae.