Phosphorylation and subcellular redistribution of pleckstrin in human neutrophils.
Pleckstrin, originally described as a major substrate of protein kinase C (PKC) in platelets, was found to be highly expressed in human neutrophils (intracellular concentration, approximately 15 microM). As PKC isoforms play an important role in mediating neutrophil antimicrobial responses, we studied the regulation of pleckstrin phosphorylation in response to inflammatory stimuli. Following treatment of neutrophils with FMLP, 12-O-tetradecanoylphorbol-13-acetate, or opsonized zymosan, pleckstrin was rapidly phosphorylated, which resulted in a shift in its electrophoretic mobility. Several lines of evidence suggest that pleckstrin is phosphorylated in part by a nonconventional PKC following stimulation by FMLP: 1) chelation of intracellular Ca2+ had only a partial inhibitory effect; 2) diacylglycerol kinase inhibitors shortened the duration of phosphorylation, while the phosphatidic acid phosphohydrolase antagonist propranolol extended it; and 3) wortmannin and erbstatin blocked the phosphorylation of pleckstrin. These results suggest that nonconventional PKC isoforms, possibly delta or zeta, mediate the phosphorylation of pleckstrin. Both PKCdelta and -zeta are expressed in human neutrophils. Increased association of pleckstrin with both microsomes and with the cytoskeleton was observed in stimulated cells. These findings suggest that phosphorylation by nonconventional PKC isoforms induces a conformational change in pleckstrin that promotes its interaction with membranes and/or with the cytoskeleton. Such a translocation may serve to target proteins or lipids recognized by pleckstrin homology domains to sites where they can contribute to the microbicidal response.


Enhanced intra-switch region recombination during immunoglobulin class switch recombination in 53BP1-/- B cells.
Immunoglobulin class switch recombination (CSR) is initiated by activation-induced cytidine deaminase (AID), an enzyme that deaminates cytidine residues in single-stranded DNA. U:G mismatches created by AID are processed to produce lesions that recruit and activate DNA damage response proteins including Ataxia-telangiectasia mutated (ATM), histone H2AX, Nijmegen breakage syndrome 1 (Nbs1), and p53 binding protein 1 (53BP1). Among these proteins, absence of 53BP1 produces the most severe impairment of class switching. Here, we demonstrate that AID is targeted normally to switch region DNA and that intra-switch region recombination is enhanced in 53BP1-/- B cells. In addition, Smicro-Sgamma1 switch region junctions cloned from 53BP1-/- B cells show unusual insertions suggestive of failed class switching. Our data are consistent with a role for 53BP1 in stabilizing the synapsis of switch regions during CSR.


Molecular dynamic and free energy studies of primary resistance mutations in HIV-1 protease-ritonavir complexes.
To understand the basis of drug resistance of the HIV-1 protease, molecular dynamic (MD) and free energy calculations of the wild-type and three primary resistance mutants, V82F, I84V, and V82F/I84V, of HIV-1 protease complexed with ritonavir were carried out. Analysis of the MD trajectories revealed overall structures of the protein and the hydrogen bonding of the catalytic residues to ritonavir were similar in all four complexes. Substantial differences were also found near the catalytic binding domain, of which the double mutant complex has the greatest impact on conformational changes of the protein and the inhibitor. The tip of the HIV-1 protease flap of the double mutant has the greater degree of opening with respect to that of the others. Additionally, the phenyl ring of Phe82 moves away from the binding pocket S1', and the conformational change of ritonavir subsite P1' consequently affects the cavity size of the protein and the conformational energy of the inhibitor. Calculations of binding free energy using the solvent continuum model were able to reproduce the same trend of the experimental inhibition constant. The results show that the resistance mutants require hydrophobic residues to maintain the interactions in the binding pocket. Changes of the cavity volume correlate well with free energy penalties due to the mutation and are responsible for the loss of drug susceptibility.


[Preliminary analysis of intensive treatment of acute lymphoblastic leukemia in children].
The analysis was carried out on 31 children with acute lymphoblastic leukaemias. In 29 of them remissions were obtained conducting later during 3 years intensive maintenance therapy associated with radiotherapy to the central nervous system (according to the method of the Memphis group). Superiority of intensive treatment in relation to more sparing treatment has been demonstrated. Therapeutic failures were observed in children with primarily poor prognosis of leukaemia, especially with blastic hyperleucocytosis. The behavior of serum immunoglobulins and especially infections occurring in children during treatment are discussed.


Coronavirus disease-2019 and heart: assessment of troponin and cardiovascular comorbidities as prognostic markers in patients hospitalized with coronavirus disease-2019 in a tertiary center in Brazil.
Our study aimed to evaluate the correlation of cardiac troponin T levels with comorbidities and in-hospital outcomes in patients with coronavirus disease-2019 in Brazil. Data from a cohort of 3,596 patients who were admitted with suspected coronavirus disease-2019 in a Brazilian tertiary center, between March and August 2020, were reviewed. A total of 2,441 (68%) patients had cardiac troponin T determined in the first 72 h of admission and were stratified into two groups: elevated cardiac troponin T (cardiac troponin T >0.014 ng/mL) and normal cardiac troponin T. Associations between troponin, comorbidities, biomarkers, and outcomes were assessed. Regression models were built to assess the association of several variables with in-hospital mortality. A total of 2,441 patients were embraced, of which 924 (38%) had normal cardiac troponin T and 1,517 (62%) had elevated cardiac troponin T. Patients with elevated cardiac troponin T were older and had more comorbidities, such as cardiovascular disease, hypertension, diabetes, arrhythmia, renal dysfunction, liver disease, stroke, cancer, and dementia. Patients with abnormal cardiac troponin T also had more altered laboratory parameters on admission (i.e., leukocytes, C-reactive protein, D-dimer, and B-type natriuretic peptide), as well as more need for intensive care unit, vasoactive drugs, mechanical ventilation, dialysis, and blood transfusion. All-cause mortality was markedly higher among patients with increased cardiac troponin T (42 vs. 16%, P<0.001). Multiple regression analysis demonstrated that in-hospital mortality was not independently associated with troponin elevation. This study showed that cardiac troponin T elevation at admission was common and associated with several comorbidities, biomarkers, and clinical outcomes in patients hospitalized with coronavirus disease-2019, but it was not an independent marker of in-hospital mortality.


Catalog of the invertebrate type specimens hosted at the Pontificia Universidad Católica del Ecuador and Escuela Politécnica Nacional natural history collections.
This work updates the invertebrate type specimen catalog published by Donoso et al. (2009). The catalog is increased by 2281 type specimens (from 454 species or subspecies) to a total of 4180 type specimens (from 770 species or subspecies) hosted at the Pontificia Universidad Católica del Ecuador and Escuela Politécnica Nacional natural history collections. The new material adds 307 holotypes, 1910 paratypes, and 64 allotypes. It provides original information from four phyla (Arthropoda, Mollusca, Nemata, and Platyhelminthes), eight classes, 21 orders, 73 families, and 156 genera. This updated catalog includes a map showing the type localities in the country, a list of the 71 new type specimens (from 23 species or subspecies) from other countries hosted at both museums, corrections to the previous catalog published by Donoso et al. (2009), and label information from each new specimen.


Molecular cloning and DNA homology of plasmid-mediated beta-lactamase genes.
Molecular cloning of DNA fragments between 1.5 and 8 kb from BamHI, EcoRI, HindIII, SalI, or Sau3A digests permitted the isolation of structural genes coding for TEM-1, ROB-1, OXA-1, OXA-3, OXA-4, OXA-5, PSE-1, PSE-2, PSE-3, PSE-4, CARB-3, CARB-4, AER-1, and LCR-1 beta-lactamases. Ampicillin-resistant clones were selected and it was confirmed that they contained the respective beta-lactamase genes by isoelectric focusing. Detailed physical maps of 14 different recombinant plasmids were constructed using 8 restriction endonucleases. Plasmid deletions and lacZ fusions were used to localize the beta-lactamase structural genes. DNA probes were constructed for the TEM-1, ROB-1, OXA-1, and PSE-1 genes. Under conditions of high stringency, hybridization was observed between the genes for TEM-1 and TEM-2 or TLE-1, OXA-1 and OXA-4, and PSE-1 and PSE-4 or CARB-3, while the ROB-1 gene probe showed no cross-hybridization. Such bla gene probes should facilitate studies of beta-lactamase molecular epidemiology.


Tracheal mucosal blood flow responses to autonomic agonists.
In lightly anesthetized adult sheep, we determined tracheal mucosal blood flow (Qtr) by measuring the steady-state uptake of dimethyl ether from a tracheal chamber created by an endotracheal tube provided with two cuffs. Qtr normalized for carotid arterial pressure [Qtr(n)] was determined before and after the exposure of the tracheal mucosa to aerosolized phenylephrine (0.25-2.0 mg), isoproterenol (0.05-0.8 mg), and methacholine (2.5-20 mg). The same doses of methacholine were also administered during the intravenous infusion of vasopressin. The measurements were repeated after intravenous pretreatment with the respective antagonists phentolamine, propranolol, and atropine. Mean +/- SE base-line Qtr(n) was 1.2 +/- 0.1 ml.min-1.mmHg-1.10(2). The autonomic antagonists had no effect on mean Qtr(n). Phenylephrine produced a dose-dependent decrease in mean Qtr(n) (-70% at the highest dose), which was blunted by phentolamine, and isoproterenol produced a dose-dependent increase in mean Qtr(n) (40% at the highest dose), which was blocked by propranolol. Methacholine failed to alter mean Qtr(n) even when Qtr was first decreased by vasopressin. We conclude that in lightly anesthetized adult sheep 1) base-line Qtr(n) is not under adrenergic or cholinergic control, 2) a locally administered alpha-adrenergic agonist decreases and beta-adrenergic agonist increases Qtr(n) via specific receptor activation, and 3) a locally administered cholinergic muscarinic agonist has no effect on Qtr(n).


Purification and characterization of nitroreductase from red alkaliphilic bacterium Aquiflexum sp. DL6.
Nitroaromatic compounds are toxic to living organisms. Most of them exhibit human mutagenic and carcinogenic potential. Biotransformation and bioremediation processes can convert these compounds into non-toxic compounds. Acclimatization of bacterial strain Aquiflexum sp. DL6 with nitro-aromatics resulted in significant induction of nitroreductase (EC 184.108.40.206). The enzyme was purified by the combination of DEAE-cellulose and Sephadex G-100 column chromatography with 80-fold purification and 22% yield. Molecular weight of purified nitroreductase was estimated to be 29 kDa by SDS-PAGE. The enzyme characteristics were explored by varying the pH and temperatures, and the optimum activity was found at pH 9.5 and 40 degrees C. It was revealed that the substrate specificity of nitroreductase of Aquiflexum sp. DL6 was wide for the most of the tested nitro-aromatic compounds. The kinetic parameters like Michaelis constant and velocity maxima were determined with o-nitrophenol and NADH as substrates.


Medical management of congenital anomalies of the kidney and urinary tract.
Renal damage in children has been found to be more congenital in origin than was previously thought. Congenital anomalies of the kidney and urinary tract (CAKUT) involve renal dysplasia, renal hypoplasia, urinary tract obstruction and vesicoureteral reflux. CAKUT are sometimes bilateral and different types often coexist. Depending on their types and severity, children with CAKUT often have varying degrees of a reduced number of nephrons at birth. CAKUTare now the leading cause of renal failure in children. Children with renal dysplasia or obstructive uropathy may have abnormal renal tubules, and tend to lose essential water and sodium in urine. This can lead to poor body growth unless they are supplemented with water and sodium. Children with severe ureteric reflux often develop urinary infection and renal scarring. Renal scarring can further increase the risk of renal failure in children who already have other CAKUTand fewer nephrons than normal. Hypertension and proteinuria may develop in children with renal dysplasia and further aggravate renal function unless they are treated. Recent advances in the understanding and management of CAKUT make it possible for children with CAKUT to grow normally, have fewer complications such as urinary infection, have longer renal survival, and survive even with end-stage renal diseases through renal replacement therapy.


Use of the alexandrite laser for treatment of seborrheic keratoses.
Seborrheic karatoses are benign lesions that are easily irritated and often cosmetically objectionable. Liquid nitrogen cryotherapy and other surgical methods are useful in treating these lesions, but are difficult to tolerate in patients who have large numbers of lesions requiring treatment. The alexandrite laser was used in one patient to quickly and efficiently destroy hundreds of seborrheic keratoses. The treatment was tolerable and excellent cosmetic results were achieved.


[Testicular seminoma in a patient with Kartagener's syndrome].
To present the association between Seminoma and Kartagener's Syndrome (KS), and its relation with infertility. We report one case of Seminoma in a patient with Kartagener's Syndrome, focussing on the possible relation between both pathologies and fertility. Bibliographic references are discussed. Orchyectomy was performed on a patient with Kartagener's Syndrome and testicular tumour. The pathology result was Seminoma. Azoospermia was obtained twice in the post operative espermiogram, not being easy to establish the implication of each pathology (KS or Seminoma) in infertility. A patient with KS and Seminoma plus azoospermia is reported, with a great outcome from the oncological point of view. Bibliographic references are described. The association between KS and Seminoma is exceptional in the literature.


Influence of DNA adsorption and DNA/cationic surfactant coadsorption on the interaction forces between hydrophobic surfaces.
The forces between hydrophobic surfaces with physisorbed DNA are markedly and irreversibly altered by exposure to DNA/cetyltrimethylammonium bromide (CTAB) mixtures. In this colloidal probe atomic force microscopy study of the interactions between a hydrophobic polystyrene particle and an octadecyltrimethylethoxysilane-modified mica surface in sodium bromide solutions, we measure distinct changes in colloidal forces depending on the existence and state of an adsorbed layer of DNA or CTAB-DNA complexes. For bare hydrophobic surfaces, a monotonically attractive approach curve and very large adhesion are observed. When DNA is adsorbed at low bulk concentrations, a long-range repulsive force dominates the approach, but on retraction some adhesion persists and DNA bridging is clearly observed. When the DNA solution is replaced with a CTAB-DNA mixture at relative low CTAB concentration, the length scale of the repulsive force decreases, the adhesion due to hydrophobic interactions greatly decreases, and bridging events disappear. Finally, when the surface is rinsed with NaBr solution, the length scale of the repulsive interaction increases modestly, and only a very tiny adhesion remains. These pronounced changes in the force behavior are consistent with CTAB-induced DNA compaction accompanied by increased DNA adsorption, both of which are partially irreversible.


Effects of vitamin E depletion and repletion on renin release from renin granules.
The present study was carried out to investigate the effects of vitamin E-deficiency and supplementation of alpha-tocopheryl acetate (TOCA) on renin release from renin granules. Male Wistar rats were fed either a control or a vitamin E-deficient diet for 4 weeks. Subsequently, the vitamin E-deficient rats received dietary supplementation of TOCA (40 mg/100 g diet) for 5 d. The renin granule fraction was prepared from the kidney cortex homogenate by a discontinuous sucrose density gradient centrifugation. The intake of vitamin E-deficient diet for 4 weeks resulted in an increased level of endogenous lipid peroxides in the renin granule fraction, accompanied by a marked decrease in alpha-tocopherol content, and led to a significant increase in the rate of renin release from the granules during incubation at 37 degrees C. These changes in alpha-tocopherol content, lipid peroxide level and renin release in the renin granule fraction were restored to the control values by dietary TOCA supplementation. Similarly, dietary supplementation of N,N'-diphenyl-p-phenylenediamine (80 mg/100 g diet), which has an antioxidative ability, suppressed the increases in lipid peroxidation and renin release due to vitamin E-deficiency, although this compound was ineffective in restoring alpha-tocopherol levels. These results suggest that vitamin E functions in maintenance of membrane integrity of renin granules by inhibiting the lipid peroxidation.


The Effect of Single-Walled Carbon Nanotubes on UDP-Glucuronosyltransferase 1A Activity in Human Liver.
Single-walled carbon nanotubes (SWCNTs) are made from rolled single graphene sheets with a diameter in the nanometer range and are potential carriers for drug delivery systems. However, their effects on uridine 5'-diphosphate-glucuronosyltransferase (UGT) 1A activities remain unclear. The present study aimed to investigate the effect of two kinds of SWCNTs (EC1.5-P- and FH-P-SWCNTs) and other nanocarbons on human UGT1A activity due to the proposed application of SWCNTs in drug and gene delivery. β-Estradiol 3-glucuronidation, which is catalyzed mainly by UGT1A1, was inhibited by 99 and 76% in the presence of 0.1 mg/mL EC1.5-P- and FH-P-SWCNTs in human liver microsomes, respectively. The observed decrease of free UGT1A1 protein in the enzyme reaction mixture suggests a higher interaction with SWCNTs, and indicates the inhibition of β-estradiol 3-glucuronidation. Imipramine N-glucuronidation, which is formed mainly by UGT1A4, was also decreased by SWCNTs. Serotonin glucuronidation, which is mainly responsible for UGT1A6, was only influenced by specific nanocarbons in human liver microsomes. The attenuation of free UGT1A6 protein was observed with SWCNTs and carbon black, indicating that UGT1A6 activity was not influenced by the direct interaction of SWCNTs. We also observed a 127% increase by FH-P-SWCNTs for propofol glucuronidation in human liver microsomes, which is catalyzed mainly by UGT1A9. The values of maximum velocity and intrinsic clearance for propofol glucuronidation in the presence of FH-P-SWCNT were 1.8- and 2.0-fold higher than those of the control in human liver microsomes. These results suggest that the effects of SWCNTs on UGT1A are different among isoforms.


Preclinical evaluation of (S)-[18F]GE387, a novel 18-kDa translocator protein (TSPO) PET radioligand with low binding sensitivity to human polymorphism rs6971.
Positron emission tomography (PET) studies with radioligands for 18-kDa translocator protein (TSPO) have been instrumental in increasing our understanding of the complex role neuroinflammation plays in disorders affecting the brain. However, (R)-[11C]PK11195, the first and most widely used TSPO radioligand has limitations, while the next-generation TSPO radioligands have suffered from high interindividual variability in binding due to a genetic polymorphism in the TSPO gene (rs6971). Herein, we present the biological evaluation of the two enantiomers of [18F]GE387, which we have previously shown to have low sensitivity to this polymorphism. Dynamic PET scans were conducted in male Wistar rats and female rhesus macaques to investigate the in vivo behaviour of (S)-[18F]GE387 and (R)-[18F]GE387. The specific binding of (S)-[18F]GE387 to TSPO was investigated by pre-treatment with (R)-PK11195. (S)-[18F]GE387 was further evaluated in a rat model of lipopolysaccharide (LPS)-induced neuroinflammation. Sensitivity to polymorphism of (S)-GE387 was evaluated in genotyped human brain tissue. (S)-[18F]GE387 and (R)-[18F]GE387 entered the brain in both rats and rhesus macaques. (R)-PK11195 blocked the uptake of (S)-[18F]GE387 in healthy olfactory bulb and peripheral tissues constitutively expressing TSPO. A 2.7-fold higher uptake of (S)-[18F]GE387 was found in the inflamed striatum of LPS-treated rodents. In genotyped human brain tissue, (S)-GE387 was shown to bind similarly in low affinity binders (LABs) and high affinity binders (HABs) with a LAB to HAB ratio of 1.8. We established that (S)-[18F]GE387 has favourable kinetics in healthy rats and non-human primates and that it can distinguish inflamed from normal brain regions in the LPS model of neuroinflammation. Crucially, we have reconfirmed its low sensitivity to the TSPO polymorphism on genotyped human brain tissue. Based on these factors, we conclude that (S)-[18F]GE387 warrants further evaluation with studies on human subjects to assess its suitability as a TSPO PET radioligand for assessing neuroinflammation.


Blood Vessel Maturation in Health and Disease and its Implications for Vascularization of Engineered Tissues.
Engineered blood vessels have often been found to be immature and unstable. Similarly, numerous pathologies such as diabetic retinopathy and cancer are characterized by highly abnormal, defective, hypervascular networks, consisting of immature, leaky, and irregular vessels with a marked loss of perivascular cell coverage. An emerging therapeutic concept in treatment of such vascular diseases and their management is the potential to normalize blood vessels by strengthening the cellular components that form the vascular network. Vessel normalization is characterized by the reduction in the number and size of immature vessels, a decrease in interstitial fluid pressure, and increase in perivascular cell coverage. Understanding the molecular and cellular defects associated with abnormal blood vessels will allow us to find appropriate treatment options that can promote normal blood vessel development. These, in turn, can be applied to improve vessel maturation in engineered tissues. In this review, we describe the major perivascular abnormalities associated with various human diseases and engineered vasculatures and the major advances in obtaining mature vasculatures for translational applications.


Nanoparticle-Drug Bioconjugate as Dual Functional Affinity Ligand for Rapid Point-of-Care Detection of Endotoxin in Water and Serum.
Endotoxin or lipopolysaccharide (LPS) is a major constituent of the Gram-negative bacterial cell wall that causes a life-threatening disorder called septicemia resulting from the unregulated activation of the innate immune system. We demonstrate a simple and robust drug-assisted dot blot bioassay for endotoxin detection that can be used right by the critically ill patients' bedside. Target LPS molecules are trapped from serum or water on glass substrates via long-chain alkyls and tagged with reporter gold nanoparticles (NPs) preconjugated to an antibiotic drug called polymyxin B sulfate (PMB). A post-silver-enhancement step enables signal visibility to the bare eye over a wide and clinically relevant concentration range of 50 fg/mL-50 ng/mL, allowing effortless diagnosis of sepsis at various stages, from early sepsis to septic shock.


Rapid kinetics and inward rectification of miniature EPSCs in layer I neurons of rat neocortex.
With the use of the whole cell patch-clamp technique combined with visualization of neurons in brain slices, we studied the properties of miniature excitatory postsynaptic currents (mEPSCs) in rat neocortical layer I neurons. At holding potentials (-50 to -70 mV) near the resting membrane potential (RMP), mEPSCs had amplitudes of 5-100 pA and were mediated mostly by alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) receptors. Amplitude histograms were skewed toward large events. An N-methyl-D-aspartate (NMDA) component was revealed by depolarization to -30 mV or by the use of a Mg2+-free bathing solution. At RMP, averaged AMPA mEPSCs had a 10-90% rise time of approximately 0.3 ms (uncorrected for instrument filtering). The decay of averaged mEPSCs was best fit by double-exponential functions in most cases. The fast, dominating component had a decay time constant of approximately 1.2 ms and comprised approximately 80% of the total amplitude. A small slow component had a decay time constant of approximately 4 ms. Positive correlations were found between rise and decay times of both individual and averaged mEPSCs, indicative of dendritic filtering. Some large-amplitude mEPSCs and spontaneous EPSCs (recorded in the absence of tetrodotoxin) had slower kinetics, suggesting a role of asynchronous transmitter release in shaping EPSCs. The amplitudes of mEPSCs were much smaller at +60 mV than at -60 mV, indicating that synaptic AMPA-receptor-mediated currents were inwardly rectifying. These results suggest that neocortical layer I neurons receive both NMDA- and AMPA-receptor-mediated synaptic inputs. The rapid decay of EPSCs appears to be largely determined by AMPA receptor deactivation. The observed rectification of synaptic responses suggests that synaptic AMPA receptors in layer I neurons may lack GluR-2 subunits and may be Ca2+ permeable.


Renal carcinomas and erythrocytosis in rats following intrarenal injection of nickel subsulfide.
In rats, an intrarenal injection of nickel subsulfide (Ni3S2) regularly caused, within 2 months, an erythrocytosis and eventually a renal carcinoma in 40 per cent of the animals. Despite their great pleomorphism, the neoplastic renal cells appeared to be of the same lineage but showed a pronounced tendency to evolve toward an anaplastic, spindle-cell variant. By electron microscopy, they exhibited varying degrees of differentiation with surface specialization, suggesting an epithelial origin. There was no indication that the erythrogenic and carcinogenic responses to Ni3S2 were interrelated as the high hemoglobin and erythrocyte values tended to retrocede with the development of the renal carcinoma. Other nickel salts and a variety of divalent metallic preparations failed to produce similar changes when administered under identical conditions. The possible mechanism of NI3S2 at the cellular level is briefly discussed.


Dissection of Closely Linked Quantitative Trait Locis Controlling Grain Size in Rice.
Grain size is a key constituent of grain weight and appearance in rice. However, insufficient attention has been paid to the small-effect quantitative trait loci (QTLs) on the grain size. In the present study, residual heterozygous populations were developed for mapping two genetically linked small-effect QTLs for grain size. After the genotyping and the phenotyping of five successive generations, qGS7.1 was dissected into three QTLs and two were selected for further analysis. The qTGW7.2a was finally mapped into a 21.10 kb interval containing four annotated candidate genes. Transcript levels assay showed that the expression of the candidates LOC_Os07g39490 and the LOC_Os07g39500 were significantly reduced in the NIL-qTGW7.2aBG1 . The cytological observation indicated that qTGW7.2a regulated the grain width through controlling the cell expansion. Using the same strategy, qTGW7.2b was fine-mapped into a 52.71 kb interval containing eight annotated candidate genes, showing a significant effect on the grain length and width with opposite allelic directions, but little on the grain weight. Our study provides new genetic resources for yield improvement and for fine-tuning of grain size in rice.


MAD2L2 inhibits colorectal cancer growth by promoting NCOA3 ubiquitination and degradation.
Nuclear receptor coactivator 3 (NCOA3) is a transcriptional coactivator that has elevated expression in multiple tumor types, including colorectal cancer (CRC). However, the molecular mechanisms that regulate the tumorigenic functions of NCOA3 in CRC remain largely unknown. In this study, we aimed to discover and identify the novel regulatory proteins of NCOA3 and explore their mechanisms of action. Immunoprecipitation (IP) coupled with mass spectrometry (IP-MS) analysis was used to detect, identify, and verify the proteins that interacted with NCOA3 in CRC cells. The biological functions of the candidate proteins and the underlying molecular mechanism were investigated in CRC cells and mouse model in vitro and in vivo. The clinical significance of NCOA3 and its interaction partner protein in CRC patients was also studied. We identified mitotic arrest deficient 2-like protein 2 (MAD2L2, also known as MAD2B or REV7), with two signal peptide sequences of LIPLK and EVYPVGIFQK, to be an interaction partner of NCOA3. Overexpression of MAD2L2 suppressed the proliferation, migration, and clonogenicity of CRC cells by inducing the degradation of NCOA3. The mechanism study showed that increased MAD2L2 expression in CRC cells activated p38, which was required for the phosphorylation of NCOA3 that led to its ubiquitination and degradation by the proteasome. Moreover, we found that MAD2L2 predicted favorable prognosis in CRC patients. We have discovered a novel role of MAD2L2 in the regulation of NCOA3 degradation and proposed that MAD2L2 serves as a tumor suppressor in CRC.


Blood transfusion during versus after cardiopulmonary bypass is associated with postoperative morbidity in neonates undergoing cardiac surgery.
Cardiac surgery on neonates for the correction of congenital heart defects is usually associated with the transfusion of packed red blood cells (PRBCs) into the cardiopulmonary bypass (CPB) circuit. We hypothesised that such transfusions of stored PRBCs directly into the arterial system may increase postoperative morbidity when compared to intravenous transfusion. In this retrospective cohort study, data from 122 consecutive neonates who received transfusions of PRBCs in the course of corrective surgery for congenital heart defects were analysed. Group assignment was according to the timing of the first transfusion: during CPB (On-CPB) or after weaning from CPB (Post-CPB). Chi Square and rank sum tests were applied to compare clinical characteristics. Times until extubation and release from the intensive care unit were analysed by Kaplan-Meier curves and by multivariate Cox regression. Transfusion of PRBCs during CPB was associated with greater 48 hour blood loss (mean±standard deviation, 86±125 versus 32±16 mL/kg, p<0.001), longer duration of mechanical ventilation (214±268 versus 99±75 h, p<0.001) and longer stay on the intensive care unit (10.9±12.1 versus 5.3±3.5 days, p<0.001). However, the groups also differed in many characteristics, such as bodyweight, complexity of surgery or preoperative haemoglobin concentration, which may also affect outcome. Yet, multivariate analyses confirmed an independent association of transfusion On-CPB with an adverse clinical outcome. Our results are consistent with the hypothesis that the transfusion of PRBCs during CPB may increase postoperative morbidity. However, due to the limitations of this retrospective analysis, further studies are needed to confirm a causal relationship between the timing of the transfusion and the clinical outcome and to elucidate putative mechanisms of such an association.


Human thymocytes become lineage committed at an early postselection CD69+ stage, before the onset of functional maturation.
Mature functional CD4 or CD8 single positive (SP) thymocytes differentiate from immature CD4+ 8+ double positive (DP) precursors through a process of positive selection and terminal differentiation. To study CD4/CD8 lineage commitment, human postselection CD69+ thymocytes were separated into distinct subpopulations based on the differential expression of CD27, CD1, and CD45RA/RO. We demonstrate that these CD69+ subpopulations represent transitional stages of a common differentiation pathway during which CD69+ thymocytes that are initially CD27- CD1+ CD45RA- will sequentially up-regulate CD27, down-regulate CD1, and eventually acquire CD45RA upon maturation. Examination of CD4 and CD8 expression on these CD69+ subsets identified an early postselection CD69+ CD27- CD4SP population that gives rise to both CD4SP and CD8SP mature T cells when cultured in mouse thymus organs. In addition, a CD4+ 8+ DP population was identified that is CD69+ and CD27+, which only gives rise to CD8SP progeny upon culture. Although these results suggest that development of CD4SP and CD8SP cells may proceed through distinct intermediates, examination of active biosynthesis of CD4 and CD8 by the various subsets demonstrated that cells that have selectively terminated CD4 synthesis are already present in the CD27- CD4SP and CD27+ DP populations before culture. These data support a model of thymocyte differentiation whereby the decision of thymocytes to differentiate into one or the other lineage occurs concomitantly with, or very soon after, acquisition of CD69 and before the cells acquire CD27, down-regulate CD1, or acquire functional properties.


Constructing Tissuelike Complex Structures Using Cell-Laden DNA Hydrogel Bricks.
Tissue engineering has long been a challenge because of the difficulty of addressing the requirements that such an engineered tissue must meet. In this paper, we developed a new "brick-to-wall" based on unique properties of DNA supramolecular hydrogels to fabricate three-dimensional (3D) tissuelike structures: different cell types are encapsulated in DNA hydrogel bricks which are then combined to build 3D structures. Signal responsiveness of cells through the DNA gels was evaluated and it was discovered that the gel permits cell migration in 3D. The results demonstrated that this technology is convenient, effective and reliable for cell manipulation, and we believe that it will benefit artificial tissue fabrication and future large-scale production.


[Injuries due to seat belts. Comments on a clinical case].
The authors report a case of associated injuries (thoracoabdominal trauma with multiple rib and sternum fractures, perforation of the small intestine, abdominal cutaneous ecchymosis) caused by a seat belt, and describe the possible seat-belt mechanisms of lesion in road accidents. Some clinical aspects are discussed; in particular the diagnostic problems concerning a seat-belt syndrome as well as the limits of effectiveness in the use of belts.


Unresponsiveness to experimental allergic encephalomyelitis in mice: replacement of suppressor cells by a soluble factor.
A soluble suppressor factor has been prepared from cells of mice rendered nonsusceptible to experimental allergic encephalomyelitis (EAE) by treatment with mouse spinal cord homogenate in incomplete Freund's adjuvant. The specific activity of this factor can be augmented by using a cell population enriched on plates coated with anti-mouse Fab and the specific antigen, mouse basic encephalitogen (MBE). The resultant suppressor factor had the same biologic activities as the cells from which it originated. Thus, it suppressed specifically the delayed-type hypersensitivity (DTH) response to MBE in vivo, and blocked in vitro the effector lymphocytes that adoptively transfer the DTH response. The suppressor factor reactivity was manifested also by the capacity to suppress the activity of macrophage migration inhibition factor produced by sensitized lymphocytes in the presence of the specific antigen MBE. The suppressor factor is antigen-specific and can bind the MBE in vitro and thus compete with its antibody binding. The most significant activity of the soluble suppressor factor is its ability to interfere with the induction of clinical EAE.


Non alcoholic fatty liver: epidemiology and natural history.
Non Alcoholic Fatty Liver Disease (NAFLD), defined as the presence of a significant amount of lipid accumulation in the liver (at least in 5% of hepatocytes), represents a challenging issue for the Hepatologists. NAFLD is not represented by a single entity, but rather by two different entities that have different natural history and evolution that range from simple fat accumulation in the liver (without any consequence), to necroinflammation, fibrosis, cirrhosis and hepatocellular carcinoma (HCC). The reason of these differences has to be found in the host characteristics and associated risk factors. Globally, its prevalence among liver diseases, and in the general population, is rising in the recent years along with its associated conditions: obesity, insulin resistance, metabolic syndrome and diabetes. This increment, together with the reported clinical conditions, may be accounted for changes in dietary habits and the increase of sedentary lifestyle. Its diffusion seems to be pandemic, given that it is beginning to affect the populations in the developing world due to the spread of Western lifestyle. This is particularly worrying in young adults and children in what seems to have become the main cause of liver disease. Even if the real rate of global incidence of NAFLD are not known, its worldwide prevalence in general population is estimated to be 20-30% in Western Countries and 5-18% in Asia and it is increasing over time. In this review we will report on the global and regional prevalence of NAFLD, the principal risk factors and the natural history of its different presentations.


Vaccination against SARS-CoV-2 infection among vulnerable and marginalised population groups in Denmark: A nationwide population-based study.
Social deprivation, psychiatric and medical disorders have been associated with increased risk of infection and severe COVID-19-related health problems. We aimed to study the rates of SARS-CoV-2 vaccination in these high-risk groups. Using health, vaccination, and administrative registers, we performed a population-based cohort study including all Danish residents aged at least 15 years, December 27, 2020, to October 15, 2021. Population groups were people experiencing: (1) homelessness, (2) imprisonment, (3) substance abuse, (4) severe mental illness, (5) supported psychiatric housing, (6) psychiatric admission, and (7) chronic medical condition. The outcome was vaccine uptake of two doses against SARS-CoV-2 infection. We calculated cumulative vaccine uptake and adjusted vaccination incidence rate ratios (IRRs) relative to the general population by sex and population group. The cohort included 4,935,344 individuals, of whom 4,277,380 (86·7%) received two doses of vaccine. Lower cumulative vaccine uptake was found for all socially deprived and psychiatrically vulnerable population groups compared with the general population. Lowest uptake was found for people below 65 years experiencing homelessness (54·6%, 95% confidence interval (CI) 53·4-55·8, p<0·0001). After adjustment for age and calendar time, homelessness was associated with markedly lower rates of vaccine uptake (IRR 0·5, 95% CI 0·5-0·6 in males and 0·4, 0·4-0·5 in females) with similar results for imprisonment. Lower vaccine uptake was also found for most of the psychiatric groups with the lower IRR for substance abuse (IRR 0·7, 0·7-0·7 in males and 0·8, 0·8-0·8 in females). Individuals with new-onset severe mental illness and, especially, those in supported psychiatric housing and with chronic medical conditions had the highest vaccine uptake among the studied population groups. Especially, socially deprived population groups, but also individuals with psychiatric vulnerability need higher priority in the implementation of the vaccination strategy to increase equity in immunization uptake. Novo Nordisk Foundation.


Shape and buckling transitions in solid-stabilized drops.
We study shape and buckling transitions of particle-laden sessile and pendant droplets that are forced to shrink in size. Monodisperse polystyrene particles were placed at the interface between water and decane at conditions that are known to produce hexagonal, crystalline arrangements on flat interfaces. As the volumes of the drops are reduced, the surface areas are likewise diminished. This effectively compresses the particle monolayer coating and induces a transition from a fluid film to a solid film. Since the particles are firmly attached to the interface by capillary forces, the shape transitions are reversible and shape/volume curves are the same for drainage and inflation. Measurements of the internal pressure of the drops reveal a strong transition in this variable as the buckling transition is approached.


[Effects of Cytokines on Early Death in Patients with Newly Diagnosed Acute Promyelocytic Leukemia].
To explore the effect of cytokine levels on early death and coagulation function of patients with newly diagnosed acute promyelocytic leukemia (APL). Routine examination was performed on 69 newly diagnosed APL patients at admission. Meanwhile, 4 ml fasting venous blood was extracted from the patients. And then the supernatant was taken after centrifugation. The concentrations of cytokines, lactate dehydrogenase (LDH) and ferritin were detected by using the corresponding kits. It was confirmed that cerebral hemorrhage was a major cause of early death in APL patients. Elevated LDH, decreased platelets (PLT) count and prolonged prothrombin time (PT) were high risk factors for early death (P <0.05). The increases of IL-5, IL-6, IL-10, IL-12p70 and IL-17A were closely related to the early death of newly diagnosed APL patients, and the increases of IL-5 and IL-17A also induced coagulation disorder in APL patients by prolonging PT (P <0.05). In newly diagnosed APL patients, ferritin and LDH showed a positive effect on the expression of IL-5, IL-10 and IL-17A, especially ferritin had a highly positive correlation with IL-5 (r =0.867) and IL-17A (r =0.841). Moreover, there was a certain correlation between these five high-risk cytokines, among which IL-5 and IL-17A (r =0.827), IL-6 and IL-10 (r =0.823) were highly positively correlated. Elevated cytokine levels in newly diagnosed APL patients increase the risk of early bleeding and death. In addition to the interaction between cytokines themselves, ferritin and LDH positively affect the expression of cytokines, thus affecting the prognosis of APL patients.


Hemoglobin, a newly recognized lipopolysaccharide (LPS)-binding protein that enhances LPS biological activity.
Cell-free hemoglobin (Hb) is a purified preparation of human hemoglobin that is being developed as a resuscitation fluid. In vivo administration of hemoglobin has resulted in significant toxicity, due in part to contamination with bacterial endotoxin (lipopolysaccharide (LPS)). To better understand this toxicity, we have studied the interaction between Hb and LPS. Mixtures of each of three different Hb preparations (cross-linked alpha alpha Hb, cross-linked carbon monoxy-alpha alpha HbCO, and non-cross-linked (native) HbAo) and LPS (Escherichia coli O26:B6 or Proteus mirabilis S1959) were examined by several independent methods for evidence of Hb.LPS complex formation. Binding assays in microtiter plates demonstrated saturable binding of LPS to immobilized Hb, with a kD of 3.1 x 10(-8) M. Binding of LPS to Hb also was demonstrated wiht a radiolabeled LPS photoaffinity probe. Ultrafiltration of Hb/LPS mixtures by 300- and 100-kDa cut-off membranes showed that the majority of LPS in these mixtures (87-97 and 64-72%, respectively) was detected in the filtrates, in contrast to the lack of filterability of LPS in the absence of Hb. Density centrifugation demonstrated that LPS co-migrated with each of the three Hbs, whereas unbound LPS had a distinctly greater sedimentation velocity than Hb or Hb.LPS complexes. Nondenaturing polyacrylamide gel electrophoresis demonstrated that in the presence of Hb, LPS migrated into the gel and co-electrophoresed with Hb, whereas LPS alone did not appreciably enter the gel. Finally, precipitation by ethanol of each of the three Hb preparations was increased in the presence of LPS compared with precipitation in the absence of LPS. Interaction of LPS with each of the three Hb preparations was also associated with altered biological activity of LPS, as shown by enhancement of LPS activation of Limulus amebocyte lysate. Therefore, our data provide several lines of independent evidence for Hb-LPS complex formation and indicated that LPS exhibited altered physical characteristics and enhanced biological activity in the presence of Hb.


Seroprevalence of bovine leptospirosis in Garanhuns Municipal District, Pernambuco State, Brazil.
The prevalence of Leptospira interrogans serovars in dairy cattle was determined by analyzing 464 serum samples from cows on 15 properties in Garanhuns municipal district, Pernambuco State, Brazil. A microscopic seroagglutination test including 12 serovars of Leptospira interrogans as antigens was used. Samples with titres 100 were considered positive. Two hundred and twenty-one (47.63%) of the samples were positive to one or more serovars. The prevalence of the serovars was hardjo (21.98%), bratislava (15.73%), castellonis (11.64%), tarassovi (10.56%), pyrogenes (1.72%), icterohaemorrhagiae (1.08%), pomona (0.86%), wolffi (0.86%), grippotyphosa (0.86%), djasiman (0.43%), canicola (0.21 %), and copenhageni (0.21%).


Biological factors as indicators of future caries.
During the last ten years the principal biological factors which have been used as indicators of future caries activity are counts of mutans streptococci and lactobacilli. The evidence that these micro-organisms play a key role in the development of dental caries has now become very strong. Methods for the identification and enumeration of mutans streptococci and lactobacilli in saliva and plaque material have been developed which are both feasible and reliable. In a few studies counts of these micro-organisms together with other factors have been related to caries incidence. The validity of the various tests show a large variation but in some studies the predictive value has been high. The best prediction of future caries activity is obtained by the combination of factors which are important contributors to the pathogenesis of dental caries.


Complement and leukocyte independent proteinuria and eicosanoid synthesis in rat membranous nephropathy.
We examined the effect of complement depletion and leukocyte depletion on an experimental model of membranous nephropathy. Nephrosis was induced in 200-gm male Sprague-Dawley rats by priming with cationic bovine gamma-globulin in adjuvant on day 1 followed by intravenous challenge with antigen starting on day 10. No naive control rats had immunofluorescent deposits in glomeruli; urine protein was less than 10 mg/24 hour and glomerular thromboxane synthesis was 658 +/- 64 ng/mg glomerular dry weight. In contrast, all rats primed and challenged with cationic bovine gamma-globulin had intense granular capillary wall deposits of rats IgG, bovine gamma-globulin, C3 and C5; severe proteinuria (183 +/- 24 mg/24 hour) was observed, associated with a 3-fold increase in glomerular thromboxane (2,393 +/- 574 ng/mg, all p less than 0.01 versus naive controls). In some rats, administration of cobra venom factor and antiserum to rat C3, starting on day 8 was used to deplete complement; hemolytic C3 and C5 were less than 2% of normal at sacrifice. These rats had IgG and bovine gamma-globulin deposits, whereas they lacked glomerular C3 or C5. Proteinuria (209 +/- 28 mg/24 hour) and glomerular thromboxane (2,087 +/- 394 ng/mg) were markedly increased compared with control, but no different from normocomplementemic rats primed and challenged with cationic bovine gamma-globulin. In other rats, depletion of leukocytes was achieved by 1,000 R x-irradiation on day 7; at sacrifice, irradiated rats had 1,270 +/- 462 wbc/microliter versus 10,375 +/- 1,652 in nephrotic rats given no other treatment, with unaltered differentials. These rats had glomerular deposits of rat IgG, bovine gamma-globulin, C3 and C5 indistinguishable from nephrotic rats with normal leukocyte counts in intensity and distribution. Proteinuria (202 +/- 30) and glomerular thromboxane (2,358 +/- 189 ng/mg) were markedly elevated compared with naive controls, but were not different from the normocomplementemic or complement-depleted groups primed and challenged with antigen. An additional control group included rats primed with ovalbumin on day 1, irradiated with 1,000 R on day 7, and challenged with cationic bovine gamma-globulin starting on day 10. This group had granular capillary wall deposits of bovine gamma-globulin, but not deposits of IgG, C3, or C5; urine protein excretion (less than 10 mg/24 hours) and glomerular thromboxane synthesis (613 +/- 90) were not different from naive controls. Glomerular prostaglandin E2 synthesis did not differ among the five groups.(ABSTRACT TRUNCATED AT 400 WORDS)


Theoretical study of silver-ion-mediated base pairs: the case of C-Ag-C and C-Ag-A systems.
Silver-mediated base pairs applied to DNA represent a new biomacromolecular nanomaterial useful for generating nanodevices as ion sensors. Reported herein is a full quantum chemical study devoted to give further knowledge on the electronic and energetic properties of C-Ag-C and mixed C-Ag-A mismatched base pairs. The B3LYP functional in conjunction with the dispersion effects (D3) has been applied. Single-point calculations have been also performed by using the M06-L functional. The investigation of their behavior has been extended to the duplex DNA modeled by the (dC-Ag-dC)2 and (dC-Ag-dA)2 more complex systems. The solvent effect has been taken into account by the conductor-like screening model, COSMO. In the case of mixed C-Ag-A and (dC-Ag-dA)2 systems, both the Watson-Crick and Hoogsteen arrangements have been taken into account. Results show that for (dC-Ag-dA)2 systems, the binding energies are almost double that of the corresponding values of C-Ag-A ones.


Simple method for the stretching and alignment of single adsorbed synthetic polycations.
Spin-coating of isolated positively charged macromolecules onto mica in the presence of octylamine was found to be a simple and general method of stretching and aligning the macromolecular chains. The contour length and molar mass for the stretched macromolecules can be directly measured by atomic force microscopy, which makes this method a very useful analytical tool. Moreover, the molecular height is increased by co-deposition with octylamine, which drastically improves the molecular resolution and allows even ultrathin polycations to be visualized. The reason for the key role of the octylamine is found in the formation of an ultrathin liquidlike alkylamine film, which reduces the surface energy of mica and weakens the interactions between the surface and the charged macromolecules.


Prioritizing watersheds for conservation actions in the southeastern coastal plain ecoregion.
The aim of this study was to apply and evaluate a recently developed prioritization model which uses the synoptic approach to geographically prioritize watersheds in which Best Management Practices (BMPs) can be implemented to reduce water quality problems resulting from erosion and sedimentation. The model uses a benefit-cost framework to rank candidate watersheds within an ecoregion or river basin so that BMP implementation within the highest ranked watersheds will result in the most water quality improvement per conservation dollar invested. The model was developed to prioritize BMP implementation efforts in ecoregions containing watersheds associated with the USDA-NRCS Conservation Effects Assessment Project (CEAP). We applied the model to HUC-8 watersheds within the southeastern Coastal Plain ecoregion (USA) because not only is it an important agricultural area but also because it contains a well-studied medium-sized CEAP watershed which is thought to be representative of the ecoregion. The results showed that the three HUC-8 watersheds with the highest rankings (most water quality improvement expected per conservation dollar invested) were located in the southern Alabama, northern Florida, and eastern Virginia. Within these watersheds, measures of community attitudes toward conservation practices were highly ranked, and these indicators seemed to push the watersheds to the top of the rankings above other similar watersheds. The results, visualized as maps, can be used to screen and reduce the number of watersheds that need further assessment by managers and decision-makers within the study area. We anticipate that this model will allow agencies like USDA-NRCS to geographically prioritize BMP implementation efforts.


Zinc Oxide Particles Can Cause Ovarian Toxicity by Oxidative Stress in Female Mice Model.
Zinc oxide nanoparticles (ZnO NPs) participate in all aspects of our lives, but with their wide application, more and more disadvantages are exposed. The goal of this study was to investigate the toxicity of ZnO NPs in female mice ovaries and explore its potential mechanism. In this study, adult female mice were orally exposed to 0, 100, 200, and 400 mg/kg ZnO NPs for 7 days. We explored the underlying mechanisms via the intraperitoneal injection of N-acetyl-cysteine (NAC), an inhibitor of oxidative stress, and salubrinal (Sal), an inhibitor of endoplasmic reticulum (ER) stress. The results indicated that serum estradiol and progesterone levels declined greatly with increasing ZnO NPs dosage. Hematoxylin and eosin (HE) staining revealed increased atretic follicles and exfoliated follicular granulosa cells. Moreover, at the transcriptional level, antioxidant-related genes such as Keap1 and Nrf2, and ER stress-related genes PERK, eIF2α, and ATF4 were markedly upregulated. In addition, the expression of Caspase12, Caspase9, and Caspase3, which are genes related to apoptosis, was also upregulated in all ZnO NPs treatment groups. Serum malondialdehyde (MDA) content was remarkably up-regulated, whereas superoxide dismutase (SOD) activity was down-regulated. The 400 mg/kg ZnO NPs treatment group suffered the most substantial harm. However, ovarian damage was repaired when NAC and Sal were added to this group. ZnO NPs had toxic effects on the ovary of female mice, which were due to oxidative stress, ER stress, and the eventual activation of apoptosis.


Some effects of products from necrotic regions of tumours on the in vitro migration of cancer and peritoneal exudate cells.
The invasion of normal tissues by cancer cells and the infiltration of tumours by macrophages to some degree involves active translatory movements by these cells. As necrosis is a common occurrence in solid tumours, we have studied the interactions of saline extracts from the necrotic regions of rat Walker-256 tumours and mouse Gardner lymphosarcomas on the transmembrane, in vitro migration of Walker and Gardner cancer cells, and rat and mouse peritoneal macrophages. Necrotic extracts enhanced cell migration independently of chemotactic activity, an effect which was partially reduced by Trasylol, an inhibitor of neutral proteases. Rat liver lysosomal preparations which contain lower levels of neutral proteases than necrotic extract, inhibited or had no effects on cell migration, and the lysosomal stabilizer hydrocortisone did not inhibit the action of necrotic extracts. The results indicate that necrosis may enhance the migration of cancer cells out of tumours and the migration of macrophages into them. The extracts act partially through their neutral protease content. In contrast to the enhancement of cell detachment by necrotic extracts, which previous work has shown to be partially mediated by their indirect effect in causing lysosomal labilization and which is partially inhibited by hydrocortisone, this indirect mechanism is not demonstrable in the enhancement of cell migration. The results indicate that the consequences of necrosis are worthy of consideration in attempts at understanding the biology of solid tumours.


Association between lower urinary tract symptoms and serum levels of sex hormones in men.
Despite the well-known association between lower urinary tract symptoms (LUTS) and male sexual dysfunction, few data are available that describe the relationship between LUTS and sex hormones. The aim of this study was to investigate a possible association between the severity of LUTS and the serum levels of sex hormones in men. A total of 182 randomly selected men attending our general urology clinic were recruited. All participants completed the International Prostate Symptom Score (IPSS), and their sera were evaluated for serum levels of prostate-specific antigen, total and free testosterone, dehydroepiandrosterone sulfate (DHEA-S), luteinizing hormone, follicle-stimulating hormone, prolactin, and estradiol. Storage symptoms were evaluated by the sum of IPSS questions 2, 4, and 7. Voiding symptoms were evaluated by the sum of IPSS questions 3, 5, and 6. The relationship between the IPSSs and serum hormone levels was determined. On univariate analysis, the total IPSS was significantly associated with age, DHEA-S, and free testosterone. Storage symptoms were significantly associated with age, DHEA-S, free testosterone, and prostate-specific antigen. Voiding symptoms were significantly associated with age and DHEA-S. On multivariate linear regression analysis, only age was significantly associated with LUTS. However, within the subgroup of men 65-82 years old, DHEA-S was significantly associated with the total IPSS and storage symptoms. In our study, the severity of LUTS was not associated with serum levels of sex hormones in men. However, in older men, storage symptoms could be affected by the serum DHEA-S level.


Anakinra for palmoplantar pustulosis: results from a randomized, double-blind, multicentre, two-staged, adaptive placebo-controlled trial (APRICOT).
Palmoplantar pustulosis (PPP) is a rare, debilitating, chronic inflammatory skin disease that affects the hands and feet. Clinical, immunological and genetic findings suggest a pathogenic role for interleukin (IL)-1. To determine whether anakinra (an IL-1 receptor antagonist) delivers therapeutic benefit in PPP. This was a randomized (1 : 1), double-blind, two-staged, adaptive, UK multicentre, placebo-controlled trial [ISCRTN13127147 (registered 1 August 2016); EudraCT number: 2015-003600-23 (registered 1 April 2016)]. Participants had a diagnosis of PPP (> 6 months) requiring systemic therapy. Treatment was 8 weeks of anakinra or placebo via daily, self-administered subcutaneous injections. Primary outcome was the Palmoplantar Pustulosis Psoriasis Area and Severity Index (PPPASI) at 8 weeks. A total of 374 patients were screened; 64 were enrolled (31 in the anakinra arm and 33 in the placebo arm) with a mean (SD) baseline PPPASI of 17·8 (10·5) and a PPP investigator's global assessment of severe (50%) or moderate (50%). The baseline adjusted mean difference in PPPASI favoured anakinra but did not demonstrate superiority in the intention-to-treat analysis [-1·65, 95% confidence interval (CI) -4·77 to 1·47; P = 0·30]. Similarly, secondary objective measures, including fresh pustule count (2·94, 95% CI -26·44 to 32·33; favouring anakinra), total pustule count (-30·08, 95% CI -83·20 to 23·05; favouring placebo) and patient-reported outcomes, did not show superiority of anakinra. When modelling the impact of adherence, the PPPASI complier average causal effect for an individual who received ≥ 90% of the total treatment (48% in the anakinra group) was -3·80 (95% CI -10·76 to 3·16; P = 0·285). No serious adverse events occurred. No evidence for the superiority of anakinra was found. IL-1 blockade is not a useful intervention for the treatment of PPP.


Relevance of Pathological Complete Response after Neoadjuvant Therapy for Breast Cancer.
Breast cancer is a heterogeneous disease, and the different biological subtypes have different prognostic impacts. Neoadjuvant trials have recently become popular as they offer several advantages compared to traditional adjuvant trials. Studies have shown that patients who achieve pathological complete response (pCR) after neoadjuvant treatment have a better long-term outcome. Consequently, increasing the rate of pCR became the end point of neoadjuvant trials with the expectation of translation into improved survival. However, the definition of pCR has lacked uniformity, and the prognostic impact of achievement of pCR on survival in different breast cancer subtypes is uncertain. In this review, we present the controversies associated with the use of pCR as an end point in neoadjuvant trials.


Gemcitabine in treating patients with refractory or relapsed multiple myeloma.
Patients with refractory or relapsed multiple myeloma are considered to have a very poor prognosis, and new regimens are needed to improve the outcome. Gemcitabine, a nucleoside antimetabolite, is an analog of deoxycytidine which mainly inhibits DNA synthesis through interfering with DNA chain elongation and depleting deoxynucleotide stores, resulting in gemcitabine-induced cell death. Here we performed a systemic analysis to evaluate gemcitabine based chemotherapy as salvage treatment for patients with refractory and relapsed multiple myeloma. Clinical studies evaluating the impact of gemcitabine based regimens on response and safety for patients with refractory and relapsed multiple myeloma were identified by using a predefined search strategy. Pooled response rate (RR) of treatment were calculated. In gemcitabine based regimens, 3 clinical studies which including 57 patients with refractory and relapsed multiple myeloma were considered eligible for inclusion. Systemic analysis suggested that, in all patients, pooled RR was 15.7% (9/57) in gemcitabine based regimens. Major adverse effects were hematologic toxicity, including grade 3 or 4 anemia, leucopenia and thrombocytopenia i. No treatment related death occurred with gemcitabine based treatment. This systemic analysis suggests that gemcitabine based regimens are associated with mild activity with good tolerability in treating patients with refractory or relapsed multiple myeloma.


Critical care management of the patient with cirrhosis awaiting liver transplant in the intensive care unit.
Patients with cirrhosis who are awaiting liver transplantation (LT) are at high risk for developing critical illnesses. Current liver allocation policies that dictate a "sickest first" approach coupled with a mismatch between need and availability of organs result in longer wait times, and thus, patients are becoming increasingly ill while awaiting organ transplantation. Even patients with well-compensated cirrhosis may suffer acute deterioration; the syndrome of acute-on-chronic liver failure (ACLF) results in multisystem organ dysfunction and a marked increase in associated short-term morbidity and mortality. For patients on transplant waiting lists, the development of multisystem organ failure may eliminate candidacy for transplant by virtue of being "too sick" to safely undergo transplantation surgery. The goals of intensive care management of patients suffering ACLF are to rapidly recognize and treat inciting events (eg, infection and bleeding) and to aggressively support failing organ systems to ensure that patients may successfully undergo LT. Management of the critically ill ACLF patient awaiting transplantation is best accomplished by multidisciplinary teams with expertise in critical care and transplant medicine. Such teams are well suited to address the needs of this unique patient population and to identify patients who may be too ill to proceed to transplantation surgery. The focus of this review is to identify the common complications of ACLF and to describe our approach management in critically ill patients awaiting LT in our centers. Liver Transplantation 23 1465-1476 2017 AASLD.


Pyogenic Hepatic Abscess in an Immunocompetent Patient With Poor Oral Health and COVID-19 Infection.
Pyogenic hepatic abscess (PHA) is a collection of pus in the liver caused by bacterial infection of the liver parenchyma. PHA is more common in immunosuppressed individuals and those with diabetes mellitus, cancer, and liver transplant. We present a rare case of PHA with Fusobacterium nucleatum in an immunocompetent patient with poor oral health, history of diverticulitis, and recent COVID-19 infection whose only symptoms were chest pain and a 4-week history of fever and malaise. The source of infection in this patient was likely dental caries and periodontal disease, with COVID-19 infection playing a role as a red herring in this patient's disease progression, delaying diagnosis. Diagnosis and treatment of PHA must be prompt with drainage and empiric anaerobic coverage followed by a more tailored antibiotic regimen if indicated by culture and further drainage if indicated by computed tomography.


Map-based cloning and expression analysis of BMR-6 in sorghum.
Brown midrib mutants in sorghum are associated with reduced lignin content and increased cell wall digestibility. In this study, we characterized a bmr-6 sorghum mutant, which shows reddish pigment in the midrib and stem after the fifth-leaf stage. Compared to wild type, Kalson lignin content of bmr-6 is decreased significantly. We used histological analysis to determine that the mutant exhibited a modified pattern of lignin staining and found an increased polysaccharide content. We cloned BMR-6 gene, a gene encoded a cinnamyl alcohol dehydrogenase (CAD), using a map-based cloning approach. Genetic complementation confirmed that CAD is responsible for the BMR-6 phenotype. BMR-6 gene was expressed in all tested sorghum tissues, with the highest being in midrib and stem. Transient expression assays in Nicotiana benthamiana leaves demonstrated cytomplasmic localization of BMR-6. We found that the expression level of bmr-6 was significantly decreased in the mutant but expression of SbCAD3 and SbCAD5 were significantly increased. Our results indicate that BMR-6 not only affects the distribution of lignin but also the biosynthesis of lignin in sorghum.


Arabidopsis POT1A interacts with TERT-V(I8), an N-terminal splicing variant of telomerase.
Chromosome integrity is maintained via the actions of ribonucleoprotein complexes that can add telomeric repeats or can protect the chromosome end from being degraded. POT1 (protection of telomeres 1), a class of single-stranded-DNA-binding proteins, is a regulator of telomeric length. The Arabidopsis genome contains three POT1 homologues: POT1A, POT1B and POT1C. Using yeast two-hybrid assays to identify components of a potential POT1A complex, we retrieved three interactors: the N-terminus of the telomerase, a protein kinase and a plant-specific protein. Further analysis of the interaction of POT1 proteins with telomerase showed that this interaction is specific to POT1A, suggesting a specific role for this paralogue. The interaction is specific to the N-terminal region of the telomerase, which can be encoded by splicing variants. This interaction indicates possible mechanisms for telomerase regulation by alternative splicing and by POT1 proteins.


The osmotic activation of transporter ProP is tuned by both its C-terminal coiled-coil and osmotically induced changes in phospholipid composition.
Transporter ProP of Escherichia coli (ProPEc) senses extracellular osmolality and mediates osmoprotectant uptake when it is rising or high. A replica of the ProPEc C terminus (Asp468-Arg497) forms an intermolecular alpha-helical coiled-coil. This structure is implicated in the osmoregulation of intact ProPEc, in vivo. Like that from Corynebacterium glutamicum (ProPCg), the ProP orthologue from Agrobacterium tumefaciens (ProPAt) sensed and responded to extracellular osmolality after expression in E. coli. The osmotic activation profiles of all three orthologues depended on the osmolality of the bacterial growth medium, the osmolality required for activation rising as the growth osmolality approached 0.7 mol/kg. Thus, each could undergo osmotic adaptation. The proportion of cardiolipin in a polar lipid extract from E. coli increased with extracellular osmolality so that the osmolality activating ProPEc was a direct function of membrane cardiolipin content. Group A ProP orthologues (ProPEc, ProPAt) share the C-terminal coiled-coil domain and were activated at low osmolalities. Like variant ProPEc-R488I, in which the C-terminal coiled-coil is disrupted, ProPEc derivatives that lack the coiled-coil and Group B orthologue ProPCg required a higher osmolality to activate. The amplitude of ProPEc activation was reduced 10-fold in its deletion derivatives. The coiled-coil structure is not essential for osmotic activation of ProP per se. However, it tunes Group A orthologues to osmoregulate over a low osmolality range. Coiled-coil lesions may impair both coiled-coil formation and interaction of ProPEc with amplifier protein ProQ. Cardiolipin may contribute to ProP adaptation by altering bulk membrane properties or by acting as a ProP ligand.


Massive Boson Superradiant Instability of Black Holes: Nonlinear Growth, Saturation, and Gravitational Radiation.
We study the superradiant instability of a massive boson around a spinning black hole in full general relativity without assuming spatial symmetries. We focus on the case of a rapidly spinning black hole in the presence of a vector boson with a Compton wavelength comparable to the black hole radius, which is the regime where relativistic effects are maximized. We follow the growth of the boson cloud through superradiance into the nonlinear regime as it spins down the black hole, reaches a maximum energy, and begins to dissipate through the emission of gravitational waves. We find that the superradiant instability can efficiently convert a significant fraction of a black hole's rotational energy into gravitational radiation.


Genomewide association analysis of sow lactation performance traits in lines of Yorkshire pigs divergently selected for residual feed intake during grow-finish phase.
Lactation is an economically and biologically important phase in the life cycle of sows. Short generation intervals in nucleus herds and low heritability of traits associated with lactation along with challenges associated with collecting accurate lactation performance phenotypes emphasize the importance of using genomic tools to examine the underlying genetics of these traits. We report the first genomewide association study (GWAS) on traits associated with lactation and efficiency in 2 lines of Yorkshire pigs that were divergently selected for residual feed intake during grow-finish phase. A total of 862 farrowing records from 2 parities were analyzed using a Bayesian whole genome variable selection model (Bayes B) to locate 1-Mb regions that were most strongly associated with each trait. The GWAS was conducted separately for parity 1 and 2 records. Marker-based heritabilities ranged from 0.03 to 0.39 for parity 1 traits and from 0.06 to 0.40 for parity 2 traits. For all traits studied, around 90% of genetic variance came from a large number of genomic regions with small effects, whereas genomic regions with large effects were found to be different for the same trait measured in parity 1 and 2. The highest percentage of genetic variance explained by a 1-Mb window for each trait ranged from 0.4% for feed intake during lactation to 4.2% for back fat measured at farrowing in parity 1 sows and from 0.2% for lactation feed intake to 5.4% for protein mass loss during lactation in parity 2 sows. A total of thirteen 1-Mb nonoverlapping windows were found to explain more than 1.5% of genetic variance for either a single trait or across multiple traits. These 1-Mb windows were on chromosomes 2, 3, 6, 7, 8, 11, 14, 15, 17, and 18. The major positional candidate genes within 1 Mb upstream and downstream of these windows were , (SSC2), (SSC6) (SSC7), (SSC8), (SSC11), (SSC14), (SSC17). Further validation studies on larger populations are required to validate these findings and to improve our understanding of the biology and complex genetic architecture of traits associated with sow lactation performance.


The clinical and histological response of periodontal pockets to root planing and oral hygiene.
The study evaluated, clinically and microscopically, the soft tissue response of suprabony periodontal pockets treated by root planing and oral hygiene or by oral hygiene measures alone. The participants were 22 adults beginning treatment for inflammatory periodontal disease. For each patient, three clinically similar labial or lingual pockets were scored for gingival inflammation, pocket depth, and the extent of plaque and calculus. One pocket in each patient served as a control, and a gingival biopsy was obtained to determine the pretreatment level of inflammation. The second pocket was treated by root planing and by the patient's oral hygiene measures. The third pocket was treated only by the patient's daily oral hygiene measures. Fifty-six to 63 days later, treated areas were reevaluated clinically, and biopsies were secured. Each type of therapy reduced the mean pocket depth and the incidence and severity of gingivitis; however, root planing accompanied by oral hygiene measures resulted in a statistically greater improvement than did oral hygiene measures alone.


The Biocontrol Agent and Insect Pathogen Photorhabdus luminescens Interacts with Plant Roots.
The number of sustainable agriculture techniques to improve pest management and environmental safety is rising, as biological control agents are used to enhance disease resistance and abiotic stress tolerance in crops. Here, we investigated the capacity of the Photorhabdus luminescens secondary variant to react to plant root exudates and their behavior toward microorganisms in the rhizosphere. P. luminescens is known to live in symbiosis with entomopathogenic nematodes (EPNs) and to be highly pathogenic toward insects. The P. luminescens-EPN relationship has been widely studied, and this combination has been used as a biological control agent; however, not much attention has been paid to the putative lifestyle of P. luminescens in the rhizosphere. We performed transcriptome analysis to show how P. luminescens responds to plant root exudates. The analysis highlighted genes involved in chitin degradation, biofilm regulation, formation of flagella, and type VI secretion system. Furthermore, we provide evidence that P. luminescens can inhibit growth of phytopathogenic fungi. Finally, we demonstrated a specific interaction of P. luminescens with plant roots. Understanding the role and the function of this bacterium in the rhizosphere might accelerate the progress in biocontrol manipulation and elucidate the peculiar mechanisms adopted by plant growth-promoting rhizobacteria in plant root interactions.IMPORTANCE Insect-pathogenic Photorhabdus luminescens bacteria are widely used in biocontrol strategies against pests. Very little is known about the life of these bacteria in the rhizosphere. Here, we show that P. luminescens can specifically react to and interact with plant roots. Understanding the adaptation of P. luminescens in the rhizosphere is highly important for the biotechnological application of entomopathogenic bacteria and could improve future sustainable pest management in agriculture.


Archipelago colonization by ecologically dissimilar amphibians: evaluating the expectation of common evolutionary history of geographical diffusion in co-distributed rainforest tree frogs in islands of Southeast Asia.
Widespread, co-distributed species with limited relative dispersal abilities represent compelling focal taxa for comparative phylogeography. Forest vertebrates in island archipelagos often exhibit pronounced population structure resulting from limited dispersal abilities or capacity to overcome marine barriers to dispersal. The exceptionally diverse Old World tree frogs of the family Rhacophoridae have colonized the forested island archipelagos of Southeast Asia on multiple occasions, entering the islands of Indonesia and the Philippines via a "stepping stone" mode of dispersal along elongate island chains, separated by a series of marine channels. Here we evaluate the prediction that two tightly co-distributed Philippine rhacophorids colonized the archipelago during concomitant timescales and in the same, linear, "island-hopping" progression. We use a new multilocus dataset, utilize dense genetic sampling from the eastern arc of the Philippines, and we take a model-based phylogeographic approach to examining the two species for similar topological patterns of diversification, genetic structure, and timescales of diversification. Our results support some common mechanistic predictions (a general south-to-north polarity of colonization) but not others (timescale for colonization and manner and degree of lineage diversification), suggesting differing biogeographic scenarios of geographical diffusion through the archipelago and unique and idiosyncratic ecological capacities and evolutionary histories of each species.


Sleep Dysregulation Is Associated with 18F-FDG PET and Cerebrospinal Fluid Biomarkers in Alzheimer's Disease.
Sleep impairment has been commonly reported in Alzheimer's disease (AD) patients. The association between sleep dysregulation and AD biomarkers has been separately explored in mild cognitive impairment (MCI) and AD patients. The present study investigated cerebrospinal-fluid (CSF) and 18F-fluoro-deoxy-glucose positron emission tomography (18F-FDG-PET) biomarkers in MCI and AD patients in order to explore their association with sleep parameters measured with polysomnography (PSG). MCI and AD patients underwent PSG, 18F-FDG-PET, and CSF analysis for detecting and correlating these biomarkers with sleep architecture. Thirty-five patients were included in the study (9 MCI and 26 AD patients). 18F-FDG uptake in left Brodmann area 31 (owing to the posterior cingulate cortex) correlated negatively with REM sleep latency (p = 0.013) and positively with REM sleep (p = 0.033). 18F-FDG uptake in the hippocampus was negatively associated with sleep onset latency (p = 0.041). Higher CSF orexin levels were associated with higher sleep onset latency (p = 0.042), Non-REM stage 1 of sleep (p = 0.031), wake after sleep onset (p = 0.028), and lower sleep efficiency (p = 0.045). CSF levels of Aβ42 correlated negatively with the wake bouts index (p = 0.002). CSF total-tau and phosphorylated tau levels correlated positively with total sleep time (p = 0.045) and time in bed (p = 0.031), respectively. Sleep impairment, namely sleep fragmentation, REM sleep dysregulation, and difficulty in initiating sleep correlates with AD biomarkers, suggesting an effect of sleep on the pathological processes in different AD stages. Targeting sleep for counteracting the AD pathological processes represents a timely need for clinicians and researchers.


Apoptosis and cytotoxicity caused by ethoxyquin and two of its salts.
In our study, we analyzed the cytotoxicity of ethoxyquin (EQ) and its two salts, ethoxyquin hydrochloride (EQ-HCL) and ethoxyquin phosphate (EQ-P). It was shown that EQ was the most cytotoxic compound (IC(50) = 0.09 mM), while the lowest cytotoxic effect was observed for EQ-P (IC(50) = 0.8 mM). The properties of ethoxyquin and its salts were also analyzed with the TUNEL method, which evaluates their ability to induce apoptosis. It was shown that EQ induced apoptosis in cultured human lymphocytes, especially at concentrations of 0.25 and 0.5 mM.


Examination of self-regulatory efficacy and pain among individuals challenged by arthritis flares.
Public health guidelines for physical activity (PA) for individuals with arthritis are 150 min/week. Self-regulatory efficacy to plan and schedule activity (SRE-SP) was greater for individuals meeting guidelines in studies when symptoms were usual. Extreme symptoms of a flare presumably challenge or block PA adherence. We found it surprising that the question of whether pain intensity and SRE-SP differ within the same person as a function of symptom severity (i.e., flare vs. no-flare) and PA level has not been addressed. Participants (N = 53) reported SRE-SP and SRE to overcome arthritis barriers (SRE-AB) during the following month, average usual and flare-pain intensity, and PA volume in the past 6 months. Mixed-model ANOVAs compared those meeting or not meeting PA guidelines in both flare and no-flare conditions. Main effects for SRE (SP and AB) were significant for within flare/no-flare comparisons (p < .001) and for SRE-SP were significant between PA groups (p < .05). Individuals meeting PA guidelines have the advantage of greater SRE-SP to motivate adherence than those of the less active. All participants' activity was less efficacious during a flare. For pain intensity, a within-subjects flare versus no-flare effect (p < .001) confirmed that flares are perceived as more of an obstacle or challenge for engaging in PA. Regardless of meeting or not meeting PA guidelines, participants reported lower SRE and higher pain intensity during a flare. PA adherence during a flare may require self-regulation of PA to be active, and in particular, to be able to meet recommended guidelines, relative to symptom severity.


Thymic carcinoma: proposal for pathological TNM and staging.
Recently, Yamakawa et al. following Masaoka's clinical staging of thymic epithelial tumors, proposed a TNM classification and staging system for thymic epithelial tumors including thymoma and thymic carcinoma. The present authors consider that division of thymomas into circumscribed types (either encapsulated or non-encapsulated but confined to within the thymus) and those invasive to adjacent organs or structures is sufficiently practical, and that a staging system is applicable to thymic carcinoma, carcinoid tumors and germ cell tumors of the anterior mediastinum, which are more malignant than thymoma. Therefore, the utility of the Yamakawa/Masaoka TNM and staging system was evaluated and a modification proposed based on experience with 16 thymic carcinomas. Although there were no cases at stage II, the survival curves obtained using the proposed modified system were more clearly separated between stages I and III or IV and between stages III and IV than the curves obtained using the Yamakawa/Masaoka system. However, the differences were not significant because of the small number of cases included. A statistically significant difference was noted between the survival curves for patients who underwent complete and incomplete surgical resection of the tumor. The utility of this proposed TNM and staging system must be evaluated by other investigators, since no cases of small cell carcinoma, lymphoepithelioma-like carcinoma, sarcomatoid carcinoma and clear cell carcinoma were included in this series, all of which are considered to have high-grade histology. An evaluation of carcinoid tumor and germ cell tumor of the anterior mediastinum must also be made.


Mineral balance in adult men: effect of four refined fibers.
Eleven men consumed a basal diet alone and with cellulose (Na-carboxymethylcellulose, locust bean gum, or karaya gum) added at 7.5 g fiber per 1000 calories for 4 wk each. Food, urine, and fecal composites were collected during the last 8 d of each feeding period. Bowel transit time was not significantly affected; however, total dry fecal weight was significantly increased after the refined fibers compared with that after the basal diet. Adding refined fibers to the basal diet did not significantly affect apparent mineral balance of calcium, magnesium, manganese, iron, copper, or zinc, with the exception of a negative mineral balance for manganese with carboxymethylcellulose. Karaya gum had a mean positive balance for all minerals tested. These results indicate that the hypocholesterolemic effect of the fibers that form gels occurs without compromising mineral balance in those subjects consuming Recommended Dietary Allowance levels of the minerals studied.


A kinase-related protein stabilizes unphosphorylated smooth muscle myosin minifilaments in the presence of ATP.
An apparent paradox in smooth muscle biology is the ability of unphosphorylated myosin to maintain a filamentous structure in the presence of ATP in vivo, whereas unphosphorylated myosin filaments are depolymerized in vitro in the presence of ATP. This suggests that additional uncharacterized factors are required for the stabilization of myosin filaments in the presence of ATP. We report here that an abundant smooth muscle protein forms sedimentable complexes with unphosphorylated smooth muscle myosin, partially reverses the depolymerizing effect of ATP on unphosphorylated myosin, and promotes the assembly of minifilaments as revealed by electron microscopy. This protein is called kinase-related protein (KRP) because it is derived from a gene within the gene for myosin light chain kinase (MLCK) and has an amino acid sequence identical to the carboxyl-terminal domain of MLCK. Consistent with the results with purified KRP, deletion of the KRP domain within MLCK results in a diminished ability of MLCK to interact with unphosphorylated myosin. KRP binds to the heavy meromyosin fragment of myosin but not to myosin rod or fragments lacking the hinge region and light chains. Altogether, these results suggest that KRP may play a critical role in stabilizing unphosphorylated myosin filaments and that the KRP domain of MLCK may be important for subcellular targeting to filaments.


Network-based approach to identify prognosis-related genes in tamoxifen-treated patients with estrogen receptor-positive breast cancer.
Tamoxifen is an estrogen receptor (ER) antagonist that is most commonly used for the treatment of ER-positive breast cancer. However, tamoxifen resistance remains a major cause of cancer recurrence and progression. Here, we aimed to identify hub genes implicated in the progression and prognosis of ER-positive breast cancer following tamoxifen treatment. Microarray data (GSE9893) for 155 tamoxifen-treated primary ER-positive breast cancer samples were obtained from the Gene Expression Omnibus database. In total, 1706 differentially expressed genes (DEGs), including 859 up-regulated and 847 down-regulated genes, were identified between relapse and relapse-free samples. Weighted correlation network analysis clustered genes into 13 modules, among which the tan and blue modules were the most significantly related to prognosis. From these two modules, we further identified and validated two prognosis-related hub genes (G-rich RNA sequence binding factor 1 (GRSF1) and microtubule-associated protein τ (MAPT)) via survival analysis based on several publicly available datasets. High expression of GRSF1 predicted poor prognosis, whereas MAPT indicated favorable outcomes in ER-positive breast cancer. Using breast cancer cell lines and tissue samples, we confirmed that GRSF1 was significantly up-regulated and MAPT was down-regulated in the tamoxifen-resistant group compared with the tamoxifen-sensitive group. The prognostic value of GRSF1 and MAPT was also verified in 48 tamoxifen-treated ER-positive breast cancer patients in our hospital. Gene set enrichment analysis (GSEA) suggested that GRSF1 was potentially involved in RNA degradation and cell cycle pathways, while MAPT was strongly linked to immune-related signaling pathways. Taken together, our findings established novel prognostic biomarkers to predict tamoxifen sensitivity, which may facilitate individualized management of breast cancer.


Changes in migraine before and after menopause in Japanese climacteric women.
Introduction We conducted a questionnaire survey of Japanese women with migraine who presented at neurology and gynecology headache clinics to assess how the frequency and severity of migraine in pre-, peri-, and postmenopausal women changed as compared with what they experienced in their 20s and 30s. Methods The study group comprised 171 women with migraine (mean age 48.5) at the time of the survey. They were requested to fill out a self-administered questionnaire. Results (1) The current frequency and severity of migraine were claimed to be higher than that in their 20s and 30s. (2) More than 50% of postmenopausal women responded that migraines were currently less frequent than in their 20s and 30s. (3) The largest proportion of postmenopausal women responded that migraine was currently less severe than during any other period. Conclusions Our survey demonstrated that there are trends toward fewer migraine attacks and milder symptoms in postmenopausal Japanese women.


Effect of heliox on albuterol delivery by metered-dose inhaler in pediatric in vitro models of mechanical ventilation.
To determine the effect of varying concentrations of heliox, a mixture of helium and oxygen, on albuterol delivery administered by metered-dose inhaler (MDI) in pediatric mechanically ventilated models. Prospective in vitro laboratory study. University-affiliated research laboratory. The lungs of a 10-kg infant and 30-kg child receiving humidified pressure-regulated volume-controlled ventilation were simulated. The infant settings were an endotracheal tube (ETT) of 4.0 mm, tidal volume of 150 ml, positive end-expiratory pressure of 2 cm H(2)O, rate of 20 breaths/minute, inspiratory time of 0.7 second; the child settings were an ETT of 6.0 mm, tidal volume of 450 ml, positive end-expiratory pressure of 2 cm H(2)O, rate of 16 breaths/minute, and inspiratory time of 0.8 second. Ten albuterol MDI canisters with chlorofluorocarbon propellants were each actuated once sequentially (total dose 1000 mug) with a commercially available aerosol holding chamber. Albuterol was collected onto a filter proximal to a lung simulator. The filter was rinsed, and concentrations were determined by high-performance liquid chromatography. In the infant model, heliox mixtures of 70:30, 60:40, and 50:50 were compared with nitrogen:oxygen (N(2):O(2)) mixtures in the same ratios. The effect of the 70:30 mixtures was also explored in a child model. Each gas mixture was tested 5 times. At all three ratios, albuterol delivery to the end of the ETT was improved with heliox compared with N(2):O(2) (approximately 7% vs 3-4%, p<0.0001, one-way analysis of variance [ANOVA] with a Bonferroni correction for multiple comparisons). No significant difference was noted in mean percentage albuterol delivery among the varying ratios of heliox studied. By two-way ANOVA, significantly greater albuterol delivery was noted with 70:30 heliox compared with 70:30 N(2):O(2) (7-8% vs 3%, p<0.0001), with no significant difference between the infant and child model (p=0.21). The gas mixture, model, and interaction of the two explained 88% of the variability in mean percentage albuterol delivery. Heliox increased albuterol delivery administered by MDI to the end of the ETT in these in vitro pediatric models of mechanical ventilation. Further studies are needed to determine if the improved albuterol delivery with heliox enhances clinical response in infants and children needing mechanical ventilation.


Prevalence, nature, and comorbidity of depressive disorders in primary care.
This article examines the prevalence, nature, and comorbidity of depressive disorders using DSM-III-R criteria among patients recruited from the waiting rooms of family physicians. A total of 1928 family practice patients completed a screening form including the Center for Epidemiologic Studies-Depression Scale (CES-D), and patients with elevated CES-D scores were oversampled for possible interviews using the Structured Clinical Interview for the DSM-III-R (SCID). In the resulting weighted sample of 425, a prevalence of 13.5% was obtained for major depression and 22.6% for all depressive disorders. Over 40% of the patients with major depressive disorder (MDD) were only mildly depressed. Gender and other demographic variables failed to distinguish depressed patients, but a variety of self-ratings did. Depression was associated with comorbid anxiety disorders and substance abuse. Results are discussed in terms of the implications of depression in primary care as a public health problem, but also in terms of some diagnostic issues, particularly the use of an impairment criterion for major depression.


Rectal endometriosis mimicking primary rectal adenocarcinoma.
Endometriosis is a benign entity defined as the presence of endometrium tissue outside of uterine cavity. It is a common disease involving peritoneum, pelvic organs, gastrointestinal tract, and so on. Diagnosis based on cytology specimen can be challenge when we encounter increased cytological atypia in the glandular epithelium, with abundant inflammatory cells and debris in the background. We presented a case of deep rectal endometriosis mimicking rectal adenocarcinoma on cytology specimen and on MRI imaging studies. The combination of endometrial glands, cellular Mullerian stroma, hemorrhage, and hemosiderin-laden macrophages are the characteristic features on cytologic specimens.


Comparison of four reconstructive methods for diaphyseal defects of the humerus after tumor resection.
The objective of the current study was to compare quantitative data on the biomechanical analysis of different techniques for fixation of intercalary bone defects of the humerus, by means of consistently applied methodology on composite models. A total of 25 humeral specimens of composite models were used. An intercalary defect was created and reconstructed using plates, intramedullary nails, external fixators and segmental prosthetic implants. The specimens were loaded under axial compression, four-point bending and torsion within the linear elastic region. Modular segmental implants and intramedullary nails were able to compensate significantly greater amounts of compressive loads compared to locking plates and external fixators. However, in flexion and torsion, the modular segmental implants and the external fixators were significantly better load-bearing devices compared to the intramedullary nails and plates. Early mobilization of the upper limb in patients with diaphyseal bone defects of the humerus could probably be more safe and tolerable when reconstructed with modular segmental implants.


Fat Grafting in Facial Palsy: A Secondary Revision Technique to Improve the Facial Aesthetics.
We report our experience of using autologous fat grafting (AFG) as an adjunct to dynamic and static facial reanimation surgical techniques in patients with facial palsy. A consecutive series of patients with facial palsy (congenital or acquired) treated by AFG between September 2007 and October 2017 were reviewed. Multiple strategies for initial dynamic facial reanimation have been utilized. Indications for AFG included asymmetry, volume deficiency, and visible muscle tethering. Standard AFG technique was used with fat harvested from the lower abdomen or thigh and injected into multiple affected areas. Fat grafting was repeated as necessary. Two-dimensional analysis was performed using standardized pre- and postoperative photographs to assess facial symmetry. Patient, surgeon, and independent evaluator satisfaction was recorded using a five-point Likert scale (0-4). Thirty-two patients with a mean age of 43 ± 15.5 years were treated with AFG following facial reanimation. A mean of 1.7 ± 1.4 secondary procedures were performed following initial dynamic reanimation before fat grafting. The average number of AFG episodes was 2.2 ± 1.4 with a mean volume of 12.9 ± 6.0 ml. Minimal complications were seen in either the donor or the recipient sites. There was significant improvement (P ≤ 0.001) of postoperative quantitative facial symmetry following fat grafting. At one-year follow-up, surgeon, patient, and independent evaluator were mostly satisfied (3.06 ± 0.62, 3.31 ± 0.59, and 3.16 ± 0.57, respectively). We report a positive experience of correction of facial asymmetry, contour abnormality and visible muscle pull with fat transplantation following dynamic facial reanimation. The procedure has been shown to be quick and simple, with few complications.


A Severe Pharyngeal-Sensory-Ataxic Variant of Guillain-Barré Syndrome With Transient Cardiac Dysfunction and a Positive Anti-sulfatide IgM.
Guillain-Barré syndrome (GBS) is a heterogeneous group of acute immune-mediated polyradiculoneuropathy that typically presents with classic axonal or demyelinating sensory-motor type. However, there are variants of GBS with atypical presentation. We report a rare case of severe pharyngeal-sensory-ataxic variant of GBS associated with poor cardiac systolic function, elevated troponin, and positive anti-sulfatide IgM. The sensory symptom atypically started in the hands in an ascending pattern, which progressed to involve the trunk and face and, later, all limbs. It was associated with severe dysphagia, ataxia, and generalized areflexia but with preserved muscle strength in all extremities. Recognizing the atypical pattern of presentation and the ability to perform an accurate clinical localization are the utmost important initial steps in making the diagnosis. The patient showed complete recovery after immunoglobulin therapy.


Solvent-free lignin recovered by thermal-enzymatic treatment using fixed-bed reactor technology - Economic assessment.
The economic viability of producing lignin by thermal-enzymatic treatment in a high-pressure fixed-bed reactor is investigated for the first time. In this direction, different advantages (e.g. recovery of low-odor sulfur-free lignin, high process flexibility) and disadvantages (e.g. high investment for high-pressure equipment) of this technology are considered. Regarding process flexibility, four different operating modes (i.e. flow through, circulation) are investigated by varying process parameters and applications of the C5 sugar fraction. Therefore, a combined modelling approach is applied by using overall biorefinery models and a predictive fixed-bed model. At optimum process conditions, lignin can be produced at a competitive price (395 €/tLignin) when comparing to other technical lignins. This result is achieved by using the fixed-bed reactor only for thermal treatment, with the water consumption being the most important factor affecting the cost of lignin production. Compared to that the C5 sugar recovery of the pretreatment is negligible.