Bladder volume-dependent excitatory and inhibitory influence of lumbosacral dorsal and ventral roots on bladder activity in rats.
This study was undertaken to examine the role of the afferent and efferent pathways of the lumbosacral spinal nerve roots in the tonic control of bladder activity. Changes of isovolumetric bladder activity were recorded in 21 sympathectomized female rats under urethane anesthesia following transection of the dorsal (DRT) and ventral (VRT) lumbosacral spinal roots, and after intraperitoneal administration of hexamethonium. DRT altered the baseline intravesical pressure in a bladder volume-dependent manner in each animal. The percent change of baseline pressure after VRT following DRT was also dependent upon bladder volume. The percent change of baseline pressure after VRT alone was similarly dependent on bladder volume, but not after VRT followed by DRT. The percent change of baseline intravesical pressure (y)(-9 to +8 cm H(2)O, -56 to +46%) after DRT and VRT depended upon bladder volume (x)(y = 44.7 x -40.4) in all rats. Hexamethonium increased the amplitude of small myogenic bladder contractions after DRT and VRT. In conclusion, the bladder is tonically excited or inhibited by a local reflex pathway and by a parasympathetic reflex pathway that depends on connections with the lumbosacral spinal cord and the pelvic nerves. Both reflex mechanisms are influenced by bladder volume.


De novo expression of MECA-79 glycoprotein-determinant on developing B lymphocytes in gut-associated lymphoid tissues.
Rabbit is one of several species that depend on development of B lymphocytes in gut-associated lymphoid tissues for primary immunoglobulin-repertoire diversification. The rabbit appendix is an important site of early B-lymphocyte development. We previously reported that peripheral lymph node addressin detected by monoclonal antibody (mAb) MECA-79 played a role in recruitment of immature blood-borne B cells into neonatal rabbit appendix. Here, we report expression of an approximately 127 000 MW O-linked sulphated proteoglycan on developing B cells in appendix and Peyer's patches recognized by the mAb MECA-79. Binding of the mAb to B lymphocytes was sensitive to enzyme treatment with O-sialoglycoprotease and expression was partially inhibited by sodium chlorate, a metabolic inhibitor of sulphation. The proportions of MECA-79(+) B lymphocytes gradually increased from < 0.5% at 3 days to > 70% at 6 weeks in appendix and Peyer's patches. The proportions of MECA-79(+) B lymphocytes in spleen and peripheral blood were very low (0.5-2%). However, the MECA-79 determinant was detected on B cells in splenic germinal centres after immunization. In situ labelling of appendix cells showed that the MECA-79 determinant was expressed on fluorescein-labelled B lymphocytes that migrated from appendix into mesenteric lymph nodes. B-cell MECA-79 may be involved in interactions with T cells and/or dendritic cells. Alternatively, because we found that lymphatic endothelium in the thymus-dependent area of appendix, a site for lymphocyte exit, expressed P-selectin (CD62P), interaction of the MECA-79 determinant on B cells with CD62P may have a role in the exit of B lymphocytes from rabbit appendix.


The role of polycomb repressive complexes in biliary tract cancer.
Polycomb group proteins are major epigenetic regulators that modify histone tails. They are organized in two multi-protein complexes called polycomb repressive complex (PRC) 1 and 2. Aberrant PRC activity is known to contribute to the development and aggressiveness of many cancers. Biliary tract cancer (BTC) is a rare malignancy associated with high chemoresistance and poor clinical outcome. Here we review the role of the PRC complexes and the effects of RNAi and drug-mediated inhibition of PRC1 and PRC2 in BTC. This review gives a short overview of the composition, biochemical functions and oncogenic role of PRC complexes. We then focus on and summarize the results of current studies that address the role of PRC in BTC. Finally, we discuss options and results of therapeutic targeting of PRC in BTC. Pharmacological inhibition of the two PRC complexes seems to be a promising strategy for treatment of BTC. To date, only few studies have addressed the therapeutic effect of PRC inhibition in BTC. Therefore, it will be important to test established PRC inhibitors, such as DZNep, as well as newly developed drugs, for example, PTC209, to gain more insight into the role of the PRC complexes in BTC and potentially to develop new therapeutic strategies.


Effects of propofol and pentobarbital on ligand binding to GABAA receptors suggest a similar mechanism of action.
The GABAA receptor is allosterically modulated by a number of anesthetics and barbiturates. We have examined the effects of propofol and pentobarbital on the binding of the receptor agonist [3H]muscimol and the benzodiazepine modulators [3H]flunitrazepam and [3H]Ro15-4513 to bovine brain membranes. Both agents potentiated the binding of [3H]muscimol (5 nM), with EC50 values of 18.7 and 276 microM, respectively. The binding of [3H]muscimol is heterogeneous, suggesting the presence of both high (Kd approximately 10 nM) and low (Kd approximately 0.1-1.0 microM) affinity sites. The major effect of both propofol and pentobarbital was to increase the affinity of the lower affinity sites without changing the total binding capacity. In contrast, the steroid anesthetic alphaxalone did not affect the affinity of these sites, suggesting that this drug has distinct effects on the GABAA receptor. Propofol and pentobarbital also increased the binding of the benzodiazepine agonist [3H]flunitrazepam and decreased the binding of the inverse agonist [3H]Ro15-4513. The results of these studies demonstrate that propofol and pentobarbital modulate the binding of ligands to the GABAA receptor in a similar manner, suggesting that these drugs may have a common mechanism of action.


Presumptive primary pulmonary mast cell tumor in 2 dogs.
Two dogs were presented, each with a large solitary pulmonary mass, and cytology confirmed mast cell tumor (MCT) in each dog. One dog was euthanized following diagnosis. Thoracic computed tomography scan and exploratory thoracotomy of the second dog revealed a right pulmonary mass that would require a radical lung resection. The patient was euthanized and histopathology confirmed a poorly granulated MCT with characteristics suggestive of epitheliotropism, an uncommon finding with MCT. These represent the first reported cases of presumptive primary pulmonary MCT in dogs.


10H-3,6-Diazaphenothiazines triggered the mitochondrial-dependent and cell death receptor-dependent apoptosis pathways and further increased the chemosensitivity of MCF-7 breast cancer cells via inhibition of AKT1 pathways.
Breast cancer is one of the leading causes of death in cancer categories, followed by lung, colorectal, and ovarian among the female gender across the world. 10H-3,6-diazaphenothiazine (PTZ) is a thiazine derivative compound that exhibits many pharmacological activities. Herein, we proceed to investigate the pharmacological activities of PTZ toward breast cancer MCF-7 cells as a representative in vitro breast cancer cell model. The PTZ exhibited a proliferation inhibition (IC50 = 0.895 µM) toward MCF-7 cells. Further, cell cycle analysis illustrated that the S-phase checkpoint was activated to achieve proliferation inhibition. In vitro cytotoxicity test on three normal cell lines (HEK293 normal kidney cells, MCF-10A normal breast cells, and H9C2 normal heart cells) demonstrated that PTZ was more potent toward cancer cells. Increase in the levels of reactive oxygen species results in polarization of mitochondrial membrane potential (ΔΨm), together with suppression of mitochondrial thioredoxin reductase enzymatic activity suggested that PTZ induced oxidative damages toward mitochondria and contributed to improved drug efficacy toward treatment. The RT2 PCR Profiler Array (human apoptosis pathways) proved that PTZ induced cell death via mitochondria-dependent and cell death receptor-dependent pathways, through a series of modulation of caspases, and the respective morphology of apoptosis was observed. Mechanistic studies of apoptosis suggested that PTZ inhibited AKT1 pathways resulting in enhanced drug efficacy despite it preventing invasion of cancer cells. These results showed the effectiveness of PTZ in initiation of apoptosis, programmed cell death, toward highly chemoresistant MCF-7 cells, thus suggesting its potential as a chemotherapeutic drug.


Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?
Sandwich ELISA is an ideal antigen detection and quantification assay. Recently, it was used as the basic concept for high technology diagnostics. The specificity of the assay depends on the exclusion of detection cross-reactivity which arises from using two antibodies developed in different species. Since mice and rats are the common laboratory animals used to develop antigen specific antibodies. Therefore, the questions we addressed here were (1) can one use antigen-specific antibodies raised in mice and rats in the same assay to specifically detect/quantify antigens? and (2) which antibodies of the two rodents should be placed for capturing and for detection in the antigen-detection sandwich? Direct ELISA assay was used to assess for the specific reaction of the HRP-conjugated antibody to the target serum. First reaction was to compare between either conjugate anti-rat IgG (homologous) or anti-mouse IgG (heterologous) for the detection of rat sera IgG. Following the dilution factor optimization, the O.D. were 0.744±0.051 and 0.604±0.05, respectively (p= .004). The difference in mean O.D. of 0.14 reflected an unaccepted non-specific reaction. The second reaction was to compare between either conjugate anti-mouse IgG (homologous) and anti-rat IgG (heterologous) for the detection of mouse sera IgG. The recorded O.D. were 0.9414±0.14 and 0.317 ±0.141, respectively (p= .0002). The improved difference in mean O.D. of 0.624 reflecting a minimized cross-reaction. Our results suggest that it is possible to use both Swiss albino mice and albino rats in a single sandwich ELISA, given that the captured antibody species to be from the Swiss albino mice and the detection antibody to be from the albino rat. The described working details are limited to the source of the antibodies used in the study. However, the approach stresses on the importance of such optimization steps before making any interpretations based on the antigen detection. To our knowledge, this study is the first to cover the optimal order of the capturing and the detection antibodies in a sandwich ELISA assay. In addition to addressing the possible interfering cross-reactivity that result from using mouse and rat serum antibodies in a single assay.


On synthesizing solid polyelectrolyte microspheres from evaporating liquid marbles.
We report a simple evaporation process using liquid marbles as precursors to produce high sphericity, precisely diameter controlled polyelectrolyte microspheres. We use poly(diallyldimethylammonium chloride) (PDDA) as the test polyelectrolyte for this experimental study. We present measurements of the rate of mass loss during evaporation to demonstrate evidence of two limiting physical processes. At short times, the rate of mass loss is well described by the "D(2) law" regime, which is vapor diffusion limited. At long times, the rate of water diffusion inside the nearly solid polyelectrolyte microsphere becomes the rate-limiting step. The transition between these two limiting processes is accompanied by changes in the physical morphology inside the microsphere. We compare the estimated values of the water diffusion coefficients with the values reported in the literature to demonstrate good agreement.


The PD-1/PD-L1 axis contributes to immune metabolic dysfunctions of monocytes in chronic lymphocytic leukemia.
Immune dysfunctions in chronic lymphocytic leukemia (CLL) contribute to tumor immune escape and attenuate immune-based therapies. Monocytes/macrophages represent key components of cancer immune surveillance and effectors for antibody-mediated antitumor effects. Monocytes display an altered subset composition in CLL. Moreover, we find a changed metabolic phenotype: glucose uptake, glucose transporters and expression of glycolytic molecules are reduced. Our data establish a link between glycolytic competence and monocyte-mediated phagocytosis of tumor cells. Furthermore, we report that CLL monocytes express Bruton's tyrosine kinase (BTK). Our observations suggest that using BTK inhibitors in CLL might further aggravate the observed immune metabolic defects in monocytes. Triggering the programmed cell death-1 (PD-1) checkpoint on monocytes hampers glycolysis, phagocytosis and BTK signaling. Conversely, disrupting PD-1/PD-L1 signaling reverses these immune metabolic dysfunctions. Taken together, our findings imply a novel metabolic interplay between CLL cells and monocytes and that blocking PD-1/PD-L1 might restore metabolic together with antitumor activity of CLL monocytes/macrophages.


Polymorphonuclear proteolytic activity and milk composition change.
Relationships between Quarter Milk Cell Count (QMCC) and Tank Milk Cell Count (TMCC) with milk biochemical and technological parameters in milk and dairy products were investigated. All parameters measured were affected by the increase of TMCC and QMCC between 0 and 600 000 cells/mL. The variable effect of lactation stage which is different for different authors, is discussed. The three mechanisms, measured during the inflammation of the udder, implicated in the modification of milk quality are described (a decrease in synthesis, a decrease in the milk barrier permeability and an increase in proteolytic activities). The direct effect of plasmin in caseinolysis is well known; the specific role of the increase of somatic cells (especially PMN) in the modification of milk quality is described. Several specific proteolytic activities of PMN are described and the impact of these activities on caseinolysis is evaluated. Two hypothetical mechanisms of caseinolysis by PMN are suggested and a synthetic scheme of the role of plasmin, bacteria and somatic cells in caseinolysis is discussed.


Preclinical evaluation of NF-κB-triggered dendritic cells expressing the viral oncogenic driver of Merkel cell carcinoma for therapeutic vaccination.
Merkel cell carcinoma (MCC) is a rare but very aggressive skin tumor that develops after integration of a truncated form of the large T-antigen (truncLT) of the Merkel cell polyomavirus (MCV) into the host's genome. Therapeutic vaccination with dendritic cells (DCs) loaded with tumor antigens is an active form of immunotherapy, which intends to direct the immune system towards tumors which express the respective vaccination antigens. Cytokine-matured monocyte-derived DCs of healthy donors and MCC patients were electroporated with mRNA encoding the truncLT. To permit major histocompatibility complex (MHC) class II next to class I presentation, we used an RNA construct in which the antigen was fused to a DCLamp sequence in addition to the unmodified antigen. To further improve their immunogenicity, the DCs were additionally activated by co-transfection with the constitutively active nuclear factor (NF)-κB activator caIKK. These DCs were used to stimulate autologous CD8+ T-cells or a mixture of CD4+ and CD8+ T-cells. Then the percentage of T-cells, specific for the truncLT, was quantified by interferon (IFN)γ ELISpot assays. Both the truncLT and its DCLamp-fusion were detected within the DCs by flow cytometry, albeit the latter required blocking of the proteasome. The transfection with caIKK upregulated maturation markers and induced cytokine production. After 2-3 rounds of stimulation, the T-cells from 11 out of 13 healthy donors recognized the antigen. DCs without caIKK appeared in comparison less potent in inducing such responses. When using cells derived from MCC patients, we could induce responses for 3 out of 5 patients; however, here the caIKK-transfected DCs did not display their superiority. These results show that optimized DCs are able to induce MCV-antigen-specific T-cell responses. Therapeutic vaccination with such transfected DCs could direct the immune system against MCC.


Analysis of Staphylococcus aureus wall teichoic acid glycoepitopes by Fourier Transform Infrared Spectroscopy provides novel insights into the staphylococcal glycocode.
Surface carbohydrate moieties are essential for bacterial communication, phage-bacteria and host-pathogen interaction. Most Staphylococcus aureus produce polyribitolphosphate type Wall teichoic acids (WTAs) substituted with α- and/or β-O-linked N-acetyl-glucosamine (α-/β-O-GlcNAc) residues. GlcNAc modifications have attracted particular interest, as they were shown to govern staphylococcal adhesion to host cells, to promote phage susceptibility conferring beta-lactam resistance and are an important target for antimicrobial agents and vaccines. However, there is a lack of rapid, reliable, and convenient methods to detect and quantify these sugar residues. Whole cell Fourier transform infrared (FTIR) spectroscopy could meet these demands and was employed to analyse WTAs and WTA glycosylation in S. aureus. Using S. aureus mutants, we found that a complete loss of WTA expression resulted in strong FTIR spectral perturbations mainly related to carbohydrates and phosphorus-containing molecules. We could demonstrate that α- or β-O-GlcNAc WTA substituents can be clearly differentiated by chemometrically assisted FTIR spectroscopy. Our results suggest that whole cell FTIR spectroscopy represents a powerful and reliable method for large scale analysis of WTA glycosylation, thus opening up a complete new range of options for deciphering the staphylococcal pathogenesis related glycocode.


[Effect of apoB polymorphism on plasma lipid levels and cerebral hemorrhage in Changsha Han Chinese].
To explore the effect of apoB polymorphism on plasma lipid levels and cerebral hemorrhage in (CH) Changsha Han Chinese. One hundred thirty CH patients and 100 normal people were involved. C7673T polymorphism of apoprotein B was analyzed by PCR-restriction fragment length polymorphism (PCR-PFLP); and the triglyceride(TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL), and low density lipoprotein-cholesterol(LDL) levels were examined by oxidase method. The serum level of lipoprotein(a) was determined by immunology method. (1) Allele T frequencies of apoB C7673T in CH patients and the control group were 0.108 and 0.040, respectively. Allele T frequencies of apoB C7673T in the CH patients were significantly higher than those in the control group (P< 0.01). (2) In the CH patients, the levels of TC and LDLjC of the T/C gene type were significantly higher than those of the C/C gene type, while the levels of HDLjC of the T/C gene type were significantly lower than those of the C/C gene type (P< 0.05). ApoB C7673T polymorphism may be related to cerebral hemorrhage, and the changing blood lipid level may increase the susceptibility of CH.


Temperature-Dependent Conduction and Photoresponse in Few-Layer ReS2.
The electrical behavior and the photoresponse of rhenium disulfide field-effect transistors (FETs) have been widely studied; however, only a few works have investigated the photocurrent as a function of temperature. In this paper, we perform the electrical characterization of few-layer ReS2-based FETs with Cr-Au contacts over a wide temperature range. We exploit the temperature-dependent transfer and output characteristics to estimate the effective Schottky barrier at the Cr-Au/ReS2 interface and to investigate the temperature behavior of parameters, such as the threshold voltage, carrier concentration, mobility, and subthreshold swing. Through time-resolved photocurrent measurements, we show that the photocurrent increases with temperature and exhibits a linear dependence on the incident light power at both low and room temperatures and a longer rise/decay time at higher temperatures. We surmise that the photocurrent is affected by the photobolometric effect and light-induced desorption of adsorbates which are facilitated by the high temperature and the low pressure.


Construction of a genetic linkage map for Saccharum officinarum incorporating both simplex and duplex markers to increase genome coverage.
Saccharum officinarum L. is an octoploid with 80 chromosomes and a basic chromosome number of x = 10. It has high stem sucrose and contributes 80% of the chromosomes to the interspecific sugarcane cultivars that are grown commercially for sucrose. A genetic linkage map was developed for S. officinarum (clone IJ76-514) using a segregating population generated from a cross between Q165 (a commercial sugarcane cultivar) and IJ76-514. In total, 40 AFLP and 72 SSR primer pairs were screened across the population, revealing 595 polymorphic bands inherited from IJ76-514. These 595 markers displayed a frequency distribution different from all other sugarcane genetic maps produced, with only 40% being simplex markers (segregated 1:1). Of these 240 simplex markers, 178 were distributed on 47 linkage groups (LGs) and 62 remained unlinked. With the addition of 234 duplex markers and 80 biparental simplex markers (segregating 3:1), 534 markers formed 123 LGs. Using the multi-allelic SSR markers, repulsion phase linkage, and alignment with the Q165 linkage map, 105 of the 123 LGs could be grouped into 10 homology groups (HGs). These 10 HGs were further assigned to the 8 HGs observed in cultivated sugarcane and S. spontaneum. Analysis of repulsion phase linkage indicated that IJ76-514 is neither a complete autopolyploid nor an allopolyploid. Detection of 28 repulsion linkages that occurred between 6 pairs of LGs located in 4 HGs suggested the occurrence of limited preferential chromosome pairing in this species.


Definitions and Clinical Guidance on the Enteral Dependence Component of the Avoidant/Restrictive Food Intake Disorder Diagnostic Criteria in Children.
The aim of the current article is to offer definitive guidance on weaning children who are reliant on nasogastric/gastrostomy feeding tubes. To date, no internationally recognized definitions or principles for interventions exist, and clinics have been reliant on creating their own unique intervention criteria. To achieve the aim, 2 goals are set out within the current article. The first goal was to definitively define the process of tube weaning. To achieve this, both tube dependency and oral eating also required definitions. It is necessary for these 2 additional definitions to fully understand the process of tube weaning and the transition that the child is making within these clinical interventions. The second goal of this article was to propose a set of minimum measurement criteria within a tube weaning protocol so that different clinical practices and perspectives may be measured accurately. This would then allow outcomes from different clinical services to be compared for efficacy. The culmination of this article is a set of 5 core principles that should govern clinics that adhere to the auspices of evidence-based practice. These principles, if adopted, will provide the basis of a set of internationally recognized criteria within this field of pediatric gastroenterology.


A resected case of metachronous liver metastasis from lung cancer producing alpha-fetoprotein (AFP) and protein induced by vitamin K absence or antagonist II (PIVKA-II).
A resected case of huge liver metastasis of hepatoid adenocarcinoma of the lung is described. A 77-year-old man who presented a solitary huge liver tumor was admitted to our hospital. He had undergone right lower lobectomy of the lung for lung cancer one year before. The view of imaging studies was not a typical one of hepatocellular carcinoma. Serum levels of AFP and PIVKA-II were 334,500ng/mL and 3,890mAU/mL, respectively, and the proportion of AFP L3 was 97.9%. It was thought that they were strongly diagnostic for hepatocellular carcinoma. Extended right lobectomy of the liver was performed. Microscopically, it was poorly differentiated adenocarcinoma and diagnosed as liver metastasis from the formerly resected lung cancer. The tumor was composed of cells with both sheet-like growth and tubule formation. The neoplastic cells, in the sheet-like growth resembled hepatocellular carcinoma cells. By immunohistochemical staining with anti-AFP and anti-PIVKA-II antibodies, cancer cells of both the primary and metastatic lesions were positive. The patient eventually died of multiple liver and bone metastasis 6 months after the operation.


N-Benzyl-aniline.
The title compound, C(13)H(13)N, is an N-alkyl-ated derivative of aniline. The N atom is present in a nearly planar mol-ecular geometry (angles sums at the N atom are 358 and 359° in the two molecules of the asymmetric unit). The planes defined by the aromatic rings intersect at angles of 80.76 (4) and 81.40 (4)° in the two molecules. In the crystal, N-H⋯Cg inter-actions connect the two mol-ecules of the asymmetric unit to form infinite homodromic chains along the crystallographic b axis [N⋯π = 3.4782 (12) and 3.4642 (13) Å].


Residues and degradation dynamics of pymetrozine and chlorpyrifos in rice field ecosystem.
Field trials were conducted in Guangzhou, Nanning, and Nanjing in two consecutive years to evaluate the terminal residue levels and dissipation trend of pymetrozine and chlorpyrifos in rice ecosystem. Analyses were carried out by high-performance-liquid-chromatography for pymetrozine and gas-chromatography-mass-spectrometry for chlorpyrifos, achieved good linear relationship over range from 0.01 to 5.0 mg·kg-1 for both (r > 0.9998). Average recoveries were 86.0% to 106.0% for pymetrozine, and 79.7% to 102.3% for chlorpyrifos at the spiking levels of 0.01, 0.1 and 1.0 mg·kg-1. Half-lives of pymetrozine in paddy water, paddy soil and rice plant were 0.35-2.81, 2.69-6.95 and 1.22-3.70 days, while that of chlorpyrifos were 0.86-1.88, 3.09-6.86 and 0.58-2.84 days. Final residues of pymetrozine and chlorpyrifos in brown rice ranged from less than 0.6 to 26.0 μg·kg-1 and 14.3 to 191.6 μg·kg-1, respectively. It is recommended that 25% pymetrozine and chlorpyrifos suspension be sprayed twice at the intervals of 10 days with dosages ranging from 375 (maximum recommended dosage) to 562.5 g a.i.·ha-1 (1.5 times of the maximum recommended dosage). The rice can be harvested safely 15 days after the last application of pymetrozine and chlorpyrifos. The research results help ensures the safe application of pymetrozine and chlorpyrifos in rice ecosystem.


How effective are ventriculopleural shunts?
Thirty-eight insertions or revisions of ventriculopleural shunts were performed in 22 patients. The most common indication for pleural cerebrospinal fluid diversion was recent abdominal infection. Very minor pneumothoraces were seen frequently, but no patient became symptomatic or required treatment. Only three shunts in 2 patients were discontinued because of pleural effusions. The median survival of the entire series of ventriculopleural shunts was only 10 months. A historical control group of revised ventriculoperitoneal shunts matched for factors that have been shown to influence shunt survival was constructed from an institutional data base. The survival of simple ventriculopleural shunts in this series was not significantly different from that of simple ventriculoperitoneal shunts in patients of comparable age with a comparable recent shunt revision history (p = 0.68). Pleural cerebrospinal fluid diversion is a simple, safe, and reasonably effective option when clinical circumstances present relative or absolute contraindications to peritoneal diversion.


3-Hydroxyisobutyrate Dehydrogenase Is Involved in Both, Valine and Isoleucine Degradation in Arabidopsis thaliana.
In plants, amino acid catabolism is especially relevant in metabolic stress situations (e.g. limited carbohydrate availability during extended darkness). Under these conditions, amino acids are used as alternative substrates for respiration. Complete oxidation of the branched-chain amino acids (BCAAs) leucine, isoleucine (Ile), and valine (Val) in the mitochondria efficiently allows the formation of ATP by oxidative phosphorylation. However, the metabolic pathways for BCAA breakdown are largely unknown so far in plants. A systematic search for Arabidopsis (Arabidopsis thaliana) genes encoding proteins resembling enzymes involved in BCAA catabolism in animals, fungi, and bacteria as well as proteomic analyses of mitochondrial fractions from Arabidopsis allowed the identification of a putative 3-hydroxyisobutyrate dehydrogenase, AtHDH1 (At4g20930), involved in Val degradation. Systematic substrate screening analyses revealed that the protein uses 3-hydroxyisobutyrate but additionally 3-hydroxypropionate as substrates. This points to a role of the enzyme not only in Val but possibly also in Ile metabolism. At4g20930 knockdown plants were characterized to test this conclusion. Root toxicity assays revealed increased root growth inhibition of the mutants if cultivated in the presence of Val or Ile but not in the presence of leucine. We conclude that AtHDH1 has a dual role in BCAA metabolism in plants.


Postdeployment Fertility Challenges and Treatment in the Modern Era.
Current war-fighting environments have shifted dramatically over the past decade, and with this change, new types of injuries are afflicting American soldiers. Operative Enduring Freedom and Operation Iraqi Freedom have noted an increased use of sophisticated improvised explosive devices by adversaries. Injuries not frequently seen in previous conflict are dismounted complex blast injuries, which involve multiple proximal amputations, pelvic fractures, and extensive perineal wounds. Thus, an unforeseen consequence of the decreased mortality rate after these complex blast injuries is a new wave of U.S. service members facing the challenges of recovering from the catastrophic amputations and genitourinary injuries. New applications of sperm retrieval methods may be used in these specific populations, as they recover and wish to purse family-building goals. Spinal cord injuries, traumatic brain injuries, and considerations unique to the female soldier are explored in this review of urologic care in wounded veterans.


Comparison of quantitative atherosclerotic plaque burden from coronary CT angiography in patients with first acute coronary syndrome and stable coronary artery disease.
Coronary CTA allows characterization of non-calcified and calcified plaque and identification of high-risk plaque features. We aimed to quantitatively characterize and compare coronary plaque burden from CTA in patients with a first acute coronary syndrome (ACS) and controls with stable coronary artery disease. We retrospectively analyzed consecutive patients with non-ST-segment elevation myocardial infarction (NSTEMI) or unstable angina with a first ACS, who underwent CTA as part of their initial workup before invasive coronary angiography and age- and gender-matched controls with stable chest pain; controls also underwent CTA with subsequent invasive angiography (total n = 28). Culprit arteries were identified in ACS patients. Coronary arteries were analyzed by automated software to quantify calcified plaque (CP), noncalcified plaque (NCP), and low-density NCP (LD-NCP, attenuation <30 Hounsfield units) volumes, and corresponding burden (plaque volume × 100%/vessel volume), stenosis, remodeling index, contrast density difference (maximum percent difference in attenuation/cross-sectional area from proximal cross-section), and plaque length. ACS patients had fewer lesions (median, 1), with higher total NCP and LD-NCP burdens (NCP: 57.4% vs 41.5%; LD-NCP: 12.5% vs 8%; P ≤ .04), higher maximal stenoses (85.6% vs 53.0%; P = .003) and contrast density differences (46.1 vs 16.3%; P < .006). Per-patient CP burden was not different between ACS and controls. NCP and LD-NCP plaque burden was higher in culprit vs nonculprit arteries (NCP: 57.8% vs 9.5%; LD-NCP: 8.4% vs 0.6%; P ≤ .0003); CP was not significantly different. Culprit arteries had increased plaque lengths, remodeling indices, stenoses, and contrast density differences (46.1% vs 10.9%; P ≤ .001). Noninvasive quantitative coronary artery analysis identified several differences for ACS, both on per-patient and per-vessel basis, including increased NCP, LD-NCP burden, and contrast density difference.


Unexplained subdural hematoma in young children: is it always child abuse?
In the published reports of the developed society, subdural hematoma and/or retinal hemorrhages, in the absence of documented history of major trauma, should be considered diagnostic of child abuse. Many people used the above criteria for diagnosis, but subsequently found that retinal hemorrhages were more common in non-accidental injuries (NAI). To what extent is the proposed pathognomonic association between unexplained subdural hematoma/retinal hemorrhages and child abuse a self-fulfilling prophecy? Clinical details of nine children under 2 years with unexplained subdural hematoma admitted to Prince of Wales Hospital between 1995 and 1998 were reviewed. Four had no other physical signs of injury, five had retinal hemorrhages and one had multiple bruises over the body. Following multidisciplinary case conferences for seven children, a diagnosis of NAI was concluded in four cases, but in no case could the abuser be definitely identified. Clinical outcome was poor with seven children showing either profound disability (n = 5) or evidence of developmental delay (n = 2). In this series, NAI were not established in three of the seven cases. Did we underdiagnose child abuse in these cases? Despite a magnitude of opinion to the contrary, the issue of whether "trivial" head injury can cause subdural hemorrhages and/or retinal hemorrhages is yet unresolved. Clearly much more information on this very sensitive and serious issue is required and these data should be collected with an open mind.


Performance of the modified Richmond Agitation Sedation Scale in identifying delirium in older ED patients.
Delirium in older emergency department (ED) patients is associated with severe negative patient outcomes and its detection is challenging for ED clinicians. ED clinicians need easy tools for delirium detection. We aimed to test the performance criteria of the modified Richmond Agitation Sedation Scale (mRASS) in identifying delirium in older ED patients. The mRASS was applied to a sample of consecutive ED patients aged 65 or older by specially trained nurses during an 11-day period in November 2015. Reference standard delirium diagnosis was based on Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR) criteria, and was established by geriatricians. Performance criteria were computed. Analyses were repeated in the subsamples of patients with and without dementia. Of 285 patients, 20 (7.0%) had delirium and 41 (14.4%) had dementia. The sensitivity of an mRASS other than 0 to detect delirium was 0.70 (95% confidence interval, CI, 0.48; 0.85), specificity 0.93 (95% CI 0.90; 0.96), positive likelihood ratio 10.31 (95% CI 6.06; 17.51), negative likelihood ratio 0.32 (95% CI 0.16; 0.63). In the sub-sample of patients with dementia, sensitivity was 0.55 (95% CI 0.28; 0.79), specificity 0.83 (95% CI 0.66; 0.93), positive likelihood ratio 3.27 (95% CI 1.25; 8.59), negative likelihood ratio 0.55 (95% CI 0.28; 1.06). The sensitivity of the mRASS to detect delirium in older ED patients was low, especially in patients with dementia. Therefore its usefulness as a stand-alone screening tool is limited.


Managed care and medical education: hard cases and hard choices.
As managed care becomes more and more dominant in U.S. health care, it is coming into conflict with medical education. There are historical reasons for this: medical education traditionally excluded physicians who chose to work in health plans, and for profit managed care has tended to avoid subsidizing medical education. In order to improve the climate, three changes are necessary: medical education must understand the tense history of discord between the two; distinctions must be made between responsible and irresponsible managed care plans; and medical educators should not assume they own the moral high ground. Arrogance, a gross oversupply of physicians and especially specialists, scandals and fraud, an often callous attitude toward the poor, and other sins can be laid at medical education's door. The worse threat for both sides is that the public and payers could simply abandon both, leading to underfunding for health professions education, a society that does not trust its health care system, and the loss of superb teaching organizations. To prevent this, managed care and medical education should work together to solve several difficult problems: how to shrink the medical education infrastructure; how to report honestly the uses to which medical education funds are put; and how to identify and end irresponsible behavior on the part of health plans and medical education entities alike. If the two sides can exercise leadership in these areas, they will be able to protect and enhance the singular place of honor that medical education holds in this society.


Balance of inflammatory pathways and interplay of immune cells in the liver during homeostasis and injury.
Multiple potentially harmful stimuli challenge the liver, the chief metabolic and detoxifying organ of the human body. Due to its central anatomical location, continuous blood flow from the gastrointestinal tract through the hepatic sinusoids allows the metabolically active hepatocytes, the non-parenchymal cells and the various immune cell populations residing and patrolling in the liver to interact with antigens and microbiological components coming from the intestine. Cytokines are key mediators within the complex interplay of intrahepatic immune cells and hepatocytes, because they can activate effector functions of immune cells as well as hepatocytic intracellular signaling pathways controlling cellular homeostasis. Kupffer cells and liver-infiltrating monocyte-derived macrophages are primary sources of cytokines such as tumor necrosis factor (TNF). The liver is also enriched in natural killer (NK) and natural killer T (NKT) cells, which fulfill functions in pathogen defense, T cell recruitment and modulation of fibrogenic responses. TNF can activate specific intracellular pathways in hepatocytes that influence cell fate in different manners, e.g. pro-apoptotic signals via the caspase cascade, but also survival pathways, namely the nuclear factor (NF)-kappaB pathway. NF-kappaB regulates important functions in liver physiology and pathology. The exact dissection of the contribution of recruited and resident immune cells, their soluble cytokine and chemokine mediators and the intracellular hepatocytic response in liver homeostasis and injury could potentially identify novel targets for the treatment of acute and chronic liver disease, liver fibrosis or cirrhosis.


BIOCHEMICAL EVOLUTION ASSOCIATED WITH ANTIPREDATOR ADAPTATION IN DAMSELFLIES.
Previous studies have shown that at least two lineages of Enallagma damselflies (Odonata: Coenagrionidae) shifted from inhabiting lakes with fish as top predators to inhabiting ponds and lakes with large dragonflies as the top predators. In adapting to living with the new predator type, these lineages evolved much greater swimming speeds to avoid attacking dragonflies. In this paper, I test whether biochemical adaptations to fuel swimming arose in concert with previously identified morphological changes that increase swimming speed. I assayed the mass-specific enzyme activities of three enzymes involved in fueling strenuous activity: pyruvate kinase and lactate dehydrogenase (enzymes involved in glycolysis) and arginine kinase (the enzyme that recharges the ATP pool). Enzyme activities were determined for 14 Enallagma species from across the genus. Species that coexist with dragonfly predators had significantly higher mass-specific arginine kinase activities than species that coexist with fish, and the results of evolutionary contrasts analyses indicate that this difference between the two groups is the result of evolutionary change associated with the habitat shifts of lineages from fish lakes to dragonfly lakes. Although significant evolution was documented for lactate dehydrogenase and pyruvate kinase across the genus, evolutionary change in the activities of these enzymes was not consistent with adaptation to coexisting with dragonfly predators. Swimming bouts to avoid dragonfly predators last for only a few seconds, and the action of arginine kinase to phosphorylate ADP to make ATP will extend the duration of maximal exertion for swimming for a few seconds. However, much longer time periods (over 45 sec) are required to generate ATP via glycolysis. Therefore, selection may have favored adaptation only at the arginine kinase locus.


Ego function of morality and developing tensions that are "within".
We applaud Baumard et al.'s mutualistic account of morality but detect circularity in their articulation of how morality emerged. Contra the authors, we propose that mutualism might account for a sensitivity to convention (the ways things are done within a group) rather than for a sense of fairness. An ontogenetic perspective better captures the complexity of what it means to be moral.


Density-Dependent Effects of Amphibian Prey on the Growth and Survival of an Endangered Giant Water Bug.
Amphibian predator-insect prey relationships are common in terrestrial habitats, but amphibian larvae are preyed upon by a variety of aquatic hemipterans in aquatic habitats. This paper suggests that the survival of the nymphs of the endangered aquatic hemipteran Kirkaldyia (=Lethocerus) deyrolli (Belostomatidae: Heteroptera) is directly and indirectly affected by the abundance of their amphibian larval prey (tadpoles). Young nymphs of K. deyrolli mainly feed on tadpoles, regardless of differences in prey availability. Nymphs provided with tadpoles grow faster than nymphs provided with invertebrate prey. Therefore, tadpole consumption seems to be required to allow the nymphs to complete their larval development. In addition, the survival of K. deyrolli nymphs was greater during the period of highest tadpole density (June) than during a period of low tadpole density (July). Higher tadpole density moderates predation pressure from the water scorpion Laccotrephes japonensis (Nepidae: Heteroptera) on K. deyrolli nymphs; i.e., it has a density-mediated indirect effect. These results suggest that an abundance of tadpoles in June provides food for K. deyrolli nymphs (a direct bottom-up effect) and moderates the predation pressure from L. japonensis (an indirect bottom-up effect). An abundance of amphibian prey is indispensable for the conservation of this endangered giant water bug species.


University Student Awareness of Skin Cancer: Behaviors, Recognition, and Prevention.
Skin cancer is the most common cancer, and it often is preventable. The authors sought to evaluate behavior and knowledge regarding skin cancer among students at a Texas university. The authors recruited a diverse group of students in terms of sex, age, and ethnicity to participate in a survey regarding knowledge of skin cancer signs, use of tanning beds, and performance of self-assessment for skin cancer. Participating students could complete surveys in classrooms, at health fairs, or online via Survey Monkey. The authors examined data for the 3 variables in relation to sex, ethnicity, and age. A total of 512 responses were completed. Female students completed 371 (72.46%) surveys, and male students completed 141 (27.54%). The ethnicity of student participants was nearly evenly split among whites, African Americans, and Hispanics. Ethnicity was the most significant factor influencing the knowledge of skin cancer and behaviors to prevent it. Specifically, Hispanic and African American students possessed a lower level of skin cancer awareness. More female students than male students used tanning beds, and although use was self-reported as infrequent, the results imply that 4500 of the university's students might use tanning beds, which is concerning if extrapolated to other university student populations in Texas. Behavioral intervention is critical in reducing students' risk of skin cancer in later years, and university students must acquire knowledge to increase their awareness of skin health and to minimize their risk of developing skin cancer. Radiation therapists are uniquely positioned to share knowledge of skin cancer.


Hollow adsorption on zigzag single-walled carbon nanotubes: repulsive first-neighbor interactions.
This article completes the study of adsorption at the centers of the hexagons (hollows) making up zigzag (M, 0) single-walled carbon nanotubes, with first- and second-neighbor adsorbate-adsorbate interactions, using our established transfer matrix method. We have previously determined that the low temperature energy phase diagrams for attractive first-neighbors are the same for all M. We now show that this is not the case for repulsive first-neighbors. Therefore, our model predicts whether first-neighbors are repulsive or attractive, based solely on whether or not experimental results are M-dependent. The numerical computations are carried out for values of M < or = 18. The progressions with increasing M of the structures of the occupational configurations of the phases, and of the low temperature phase diagrams, are compared with the results known in the infinite-M limit. They strongly suggest six families of low temperature phase diagrams with M = 4, 6, 8, 10, 12, and 14, modulo 12. This can be experimentally verified, and can then be used to make further predictions on hollow adsorption on any zigzag (M, 0) nanotube.


Amine-catalyzed coupling of allenic esters to alpha,beta-unsaturated carbonyls.
Nucleophilic amines catalyze the 1,4-addition of allenic esters to alpha,beta-unsaturated carbonyl compounds. The reactions proceed under mild conditions. In addition, Baylis-Hillman products serve as appropriate electrophiles for the process such that a one-pot, three-component coupling of aldehydes, acrylates, and allenic esters may be achieved.


Functionalized Spiky Particles for Intracellular Biomolecular Delivery.
The intracellular delivery of biomolecules is of significant importance yet challenging. In addition to the conventional delivery of nanomaterials that rely on biochemical pathways, vertical nanowires have been recently proposed to physically penetrate the cell membrane, thus enabling the direct release of biomolecules into the cytoplasm circumventing endosomal routes. However, due to the inherent attachment of the nanowires to a planar 2D substrate, nanowire cell penetrations are restricted to in vitro applications, and they are incapable of providing solution-based delivery. To overcome this structural limitation, we created polyethylenimine-functionalized microparticles covered with nanospikes, namely, "spiky particles", to deliver biomolecules by utilizing the nanospikes to penetrate the cell membrane. The nanospikes might penetrate the cell membrane during particle engulfment, and this enables the bound biomolecules to be released directly into the cytosol. TiO2 spiky particles were fabricated through hydrothermal routes, and they were demonstrated to be biocompatible with HeLa cells, macrophage-like RAW cells, and fibroblast-like 3T3-L1 cells. The polyethylenimine-functionalized spiky particles provided direct delivery of fluorescent siRNA into cell cytosol and functional siRNA for gene knockdown as well as successful DNA plasmid transfection which were difficult to achieve by using microparticles without nanospikes. The spiky particles presented a unique direct cell membrane penetrant vehicle to introduce biomolecules into cell cytosol, where the biomolecules might bypass conventional endocytic degradation routes.


A second mitral valve replacement in a patient with hereditary hemorrhagic telangiectasia (Osler's disease).
A 62-year-old female with Osler's disease was admitted to our hospital because of fever and cardiac failure. The patient had undergone a mitral valve replacement (MVR) using a Carpentier-Edwards prosthetic valve 14 years earlier. A bacterial examination of arterial blood identified Streptococcus mitis. No arteriovenous malformations were detected in visceral organs. The patient underwent MVR using the same prosthetic xenograft after conservative treatment and management of repetitive epistaxis and decayed teeth. Intra- and postoperative bleeding were typical of a mitral valve reoperation. This is the first reported experience, to the best of our knowledge, of a second MVR in a patient with Osler's disease.


An Unusual Case Presentation of the May-Thurner Syndrome.
A 56-year-old woman underwent abdominoplasty with no immediate complications. She had no known bleeding history nor any relevant past surgical history. Adequate preventive measures for venous thromboembolism were performed, including sequential compression devices, good hydration, and early ambulation. At 17 days post operation, the patient presented to the emergency room complaining of left leg swelling and sharp, shooting pain radiating down her left leg. Workup in the Emergency Room revealed significant venous thrombosis involving complete occlusion of the major veins of the left leg. There were no indications of cardiopulmonary compromise. Angiography revealed an anatomical variant consistent with May-Thurner Syndrome (MTS). This variant first described in 1957 may present in up to one-quarter of patients, more commonly in young women. This case appears to the first reported of MTS occurring in association with a postoperative complication of abdominoplasty. Diagnosis and management considerations are discussed.


Immune System, Friend or Foe of Oncolytic Virotherapy?
Oncolytic viruses (OVs) are an emerging class of targeted anticancer therapies designed to selectively infect, replicate in, and lyse malignant cells without causing harm to normal, healthy tissues. In addition to direct oncolytic activity, OVs have shown dual promise as immunotherapeutic agents. The presence of viral infection and subsequently generated immunogenic tumor cell death trigger innate and adaptive immune responses that mediate further tumor destruction. However, antiviral immune responses can intrinsically limit OV infection, spread, and overall therapeutic efficacy. Host immune system can act both as a barrier as well as a facilitator and sometimes both at the same time based on the phase of viral infection. Thus, manipulating the host immune system to minimize antiviral responses and viral clearance while still promoting immune-mediated tumor destruction remains a key challenge facing oncolytic virotherapy. Recent clinical trials have established the safety, tolerability, and efficacy of virotherapies in the treatment of a variety of malignancies. Most notably, talimogene laherparepvec (T-VEC), a genetically engineered oncolytic herpesvirus-expressing granulocyte macrophage colony stimulating factor, was recently approved for the treatment of melanoma, representing the first OV to be approved by the FDA as an anticancer therapy in the US. This review discusses OVs and their antitumor properties, their complex interactions with the immune system, synergy between virotherapy and existing cancer treatments, and emerging strategies to augment the efficacy of OVs as anticancer therapies.


Translation, Association and Augmentation: Learning Cross-Modality Re-Identification From Single-Modality Annotation.
Daytime visible modality (RGB) and night-time infrared (IR) modality person re-identification (VI-ReID) is a challenging cross-modality pedestrian retrieval problem. However, training a cross-modality ReID model requires plenty of cross-modality (visible-infrared) identity labels that are more expensive than single-modality person ReID. To alleviate this issue, this paper studies unsupervised domain adaptive visible infrared person re-identification (UDA-VI-ReID) task without the reliance on any cross-modality annotation. To transfer learned knowledge from the labelled visible source domain to the unlabelled visible-infrared target domain, we propose a Translation, Association and Augmentation (TAA) framework. Specifically, the modality translator is firstly utilized to transfer visible image to infrared image, formulating generated visible-infrared image pairs for cross-modality supervised training. A Robust Association and Mutual Learning (RAML) module is then designed to exploit the underlying relations between visible and infrared modalities for label noise modeling. Moreover, a Translation Supervision and Feature Augmentation (TSFA) module is designed to enhance the discriminability by enriching the supervision with feature augmentation and modality translation. The extensive experimental results demonstrate that our method significantly outperforms current state-of-the-art unsupervised methods under various settings, and even surpasses some supervised counterparts, providing a powerful baseline for UDA-VI-ReID.


Mass spectrometric analysis of the fragments produced by cleavage and reduction of rat prolactin: evidence that the cleaving enzyme is cathepsin D.
The site(s) at which mammary tissue enzymatically cleaves rat (r) PRL, and the possibility that the cleaving activity is cathepsin D, were investigated using mass spectrometry and enzyme inhibitors. Cleavage of intact rPRL [22,566 atomic mass units (amu)] by either mammary gland-conditioned medium or cathepsin D (both at pH 3) reduced the mass of the molecule by 397 amu. Subsequent reduction of the large rPRL fragment cleaved by either method generated two fragments of 16,364 and 5808 amu. The mass of the smaller fragment is consistent with the report of Vick et al. (Biochim Biophys Acta 931: 196-204, 1987) that its amino-terminal residue is Ser149. These results indicate that both enzyme preparations cleave rPRL by excision of the tripeptide Leu-Val-Trp (mass = 397 amu) between residues 145 and 149. The ability of both enzyme preparations to cleave rPRL at pH3 was inhibited by pepstatin A but not by phenylmethane sulfonyl fluoride, and both preparations were essentially inactive at pH7. Accordingly, the PRL-cleaving activity of rat mammary tissue is probably cathepsin D.


Brief communication genotyping of Burkholderia pseudomallei revealed high genetic variability among isolates from a single population group.
Burkholderia pseudomallei is a soil dwelling Gram-negative bacteria predominates in Southeast Asia zone and the tropical part of Australia. Genetic diversity has been explored among various populations and environments worldwide. To date, little data is available on MLST profiling of clinical B. pseudomallei isolates in peninsular Malaysia. In this brief report, thirteen culture positive B. pseudomallei cases collected from a single population of Terengganu state in the Western Peninsular Malaysia and were confirmed by In-house TTS1-PCR. Isolates were subjected for multi-locus sequence typing (MLST) to explore their genotypic diversity and to investigate for possible clonal clustering of a certain sequence type. Patient's clinical information was examined to investigate for clinical correlation among the different genotypes. In spite of small sample set, MLST results indicated predictive results; considerable genotypic diversity, predominance and novelty among B. pseudomallei collected over a single geographically-located population in Malaysia. Massive genotypic heterogeneity was observed; 8 different sequence types with predominance of sequence type 54 and discovery of two novel sequence types. However, no clear pathogenomic or organ tropism clonal relationships were predicted.


Structural basis of Alzheimer β-amyloid peptide recognition by engineered lipocalin proteins with aggregation-blocking activity.
We describe the structural analysis of two Anticalin® proteins that tightly bind Aβ40, a peptide involved in the pathophysiology of Alzheimer's disease. These anticalins, US7 and H1GA, were engineered on the basis of the human lipocalin 2, thus yielding compact single-domain binding proteins as an alternative to antibodies. Albeit selected under different conditions and mutually deviating in 13 amino acid positions within the binding pocket (of 17 mutated residues in total), both crystallised anticalins recognize the same epitope in the middle of the β-amyloid peptide. In the two complexes with the Aβ40 peptide, its central part comprising residues LysP16 to LysP28 shows well defined electron density whereas the flanking regions appear structurally disordered. The compact zigzag-bend conformation which is seen in both structures may indicate a role during conversion of the soluble monomeric form into pathogenic Aβ state(s) and, thus, explain the aggregation-inhibiting effect of the anticalins. In contrast to solanezumab, which targets the same Aβ region in a different conformation, the anticalin H1GA does not show cross-reactivity with sequence-related human plasma proteins. Consequently, anticalins offer promising reagents to prevent oligomerization of Aβ peptides to neurotoxic species in vivo and their small size may enable new routes for brain delivery.


Evaluation of antibacterial in vitro activity of moxifloxacin and its effects on pulmonary clearance of Klebsiella pneumoniae in an animal experimental model.
Moxifloxacin (CAS 186286-86-8) is a recent fluoroquinolone addressed to the treatment of respiratory infections, although clinical efficacy has been shown also in the therapy of urinary tract diseases. The in vitro activity of moxifloxacin against one encapsulated strain of Klebsiella pneumoniae was determined by means of time-kill curves and by evaluation of effects of subinhibitory concentrations on bacterial morphology by means of scanning electron microscopy. Moreover, the efficacy and time requested to eradicate the same strain from lungs was determined after intratracheal infection of guinea pigs. K. pneumoniae was rapidly killed by moxifloxacin at concentrations greater than the minimum inhibitory concentration (MIC). Subinhibitory concentrations (1/4 and 1/8 of the MIC ) induced filamentation in a remarkable portion of bacteria. A progressive decrease in the bacterial content of lungs was observed during the 48-h period, with statistically significant differences compared with the control animals. In conclusion, this study provides in vivo evidence of the antimicrobial properties of moxifloxacin against K. pneumoniae.


[The significance of a carotid bruit. The preliminary experience of the OPI program. The Obiettivo prevenzione ictus (Stroke Prevention Objective)].
Carotid bruit has been correlated to occlusive atherosclerotic carotid disease and cerebrovascular ischemic disease, based on selected series or small population cohorts. The OPI programme has been accomplished to investigate the prevalence of carotid lesions in a population aged 45 to 75 and resident in the USSL (Sanitary District) n. 69, Lombardia. To establish the frequency of carotid bruit in an unselected population and the correlation of this sign to the internal carotid lesion. In this paper 16,379 people of both sexes are considered: all underwent physical examination, oriented to vascular problems, and C.W. Doppler ultrasound examination, from 1990 to March, 31st 1992. A further examination completed with color flow imaging was carried out for subjects showing either carotid bruit, or C.W. Doppler alterations, history of cerebrovascular disorders, previous vascular operations, concomitant extracarotid vascular occlusive diseases. Carotid kinking has been considered as "carotid lesion", for its attitude to cause flow alterations fitting to carotid bruit. The prevalence of carotid bruit in the considered population was 3.7%, a total of 806 carotid bruits in 612 subjects. Male to female ratio was 0.69. A strong correlation of bruit to age was recorded (p < 10(-6)): subjects over 70 years old have a five-fold probability to harbor carotid bruit than younger people (45 to 50 years old). Likewise hypertensive people have a two-fold probability of carotid bruit than normotensive (p < 10(-6)). Only 302 carotid lesions (36%) of the 864 found showed bruit, while 539 (64%) did not. Among the 31,917 unaffected carotids 504 (1.6%) had bruit and 31,413 had not (98.4%); sensitivity of bruit to carotid lesion was 35.9%, and specificity 98.4%. The probability for carotids with bruit to be affected with lesion (stenosis, occlusion or kinking) is approximately 35 times as high as for carotids without bruit (odds ratio = 34.9). Notwithstanding its low sensitivity, that prevents its use as a simple screening test for carotid lesion, the high specificity of carotid bruit makes it possible to separate a group at elevated risk of harboring carotid lesion. The absence of bruit does not rule out the presence of carotid lesion. A strong correlation to age and hypertension has been proved. Carotid bruit must be considered as a risk factor for carotid lesion, not a means to make diagnosis.


Impact of transcendental meditation on left ventricular mass in african american adolescents.
Background. An early sign of ventricular remodeling is increased left ventricular mass (LVM) which over time may lead to left ventricular hypertrophy, the strongest predictor of cardiovascular morbidity and mortality, other than advancing age. Methods. 62 (30 TM; 32 CTL) African American adolescents (age 16.2 ± 1.3 years) with high normal systolic BP were randomly assigned to either 4-month Transcendental Meditation (TM) or health education control groups. The echocardiographic-derived measure of LVM index (LVMI = LVM/ht(2.7)) was measured before and after the 4-month TM study and at 4-month followup. 2D-guided M-mode echocardiography using a Hewlett Packard 5500 echosonograph was used to determine LVMI. Results. The TM group exhibited a greater decrease in LVMI at 4-month followup compared to the CTL group (-2.6 versus +0.3 gm/ht(2.7), P < 0.04). The TM group exhibited a lesser increase in BMI at 4-month follow-up compared to the CTL group (0.2 ± 1.6 versus 1.1 ± 1.4, P < 0.03). Conclusion. These findings indicate that among a group of prehypertensive African American adolescents, 4 months of TM compared to heath education resulted in a significant decrease in LVMI, and these changes were maintained at 4-month follow-up.


Pressure sores: incidence, risk assessment and prevention.
Pressure sores have a physiological basis which, in common with other disease processes, is influenced by intrinsic and extrinsic factors unique to the individual. A comprehensive patient assessment will identify factors affecting the individual and consequently direct preventive intervention. The issue of pressure sores should be addressed at a patient, trust or regional level to ensure that preventive strategies/policies cross all healthcare boundaries. Identification of the size of the problem is the starting point upon which a preventive strategy can be built. A comprehensive strategy for pressure sore prevention has existed in the USA since 1992, and its contents demonstrate the multidisciplinary input required to develop and implement the recommendations. Nurses have a key role in the prevention of pressure sores; however, they need educational, managerial and financial support in order to function effectively within this role.


Hepatic granuloma in polymyalgia rheumatica.
Hepatic granuloma occurs rarely in the polymyalgia rheumatica syndrome. A case was presented of a 62-year-old woman with polymyalgia rheumatica, elevated concentrations of hepatic enzymes, and granuloma of the liver. Low doses of corticosteroids produced a marked improvement in the patient's condition and a return of liver function tests to normal. This is apparently the first case of polymyalgia rheumatica associated with liver granuloma that has been reported in Israel.


Time-on-task effects in children with and without ADHD: depletion of executive resources or depletion of motivation?
Children with attention-deficit/hyperactivity disorder (ADHD) are characterized by deficits in their executive functioning and motivation. In addition, these children are characterized by a decline in performance as time-on-task increases (i.e., time-on-task effects). However, it is unknown whether these time-on-task effects should be attributed to deficits in executive functioning or to deficits in motivation. Some studies in typically developing (TD) adults indicated that time-on-task effects should be interpreted as depletion of executive resources, but other studies suggested that they represent depletion of motivation. We, therefore, investigated, in children with and without ADHD, whether there were time-on-task effects on executive functions, such as inhibition and (in)attention, and whether these were best explained by depletion of executive resources or depletion of motivation. The stop-signal task (SST), which generates both indices of inhibition (stop-signal reaction time) and attention (reaction time variability and errors), was administered in 96 children (42 ADHD, 54 TD controls; aged 9-13). To differentiate between depletion of resources and depletion of motivation, the SST was administered twice. Half of the participants was reinforced during second task performance, potentially counteracting depletion of motivation. Multilevel analyses indicated that children with ADHD were more affected by time-on-task than controls on two measures of inattention, but not on inhibition. In the ADHD group, reinforcement only improved performance on one index of attention (i.e., reaction time variability). The current findings suggest that time-on-task effects in children with ADHD occur specifically in the attentional domain, and seem to originate in both depletion of executive resources and depletion of motivation. Clinical implications for diagnostics, psycho-education, and intervention are discussed.


Beta-adrenergic receptor gene polymorphisms and cardiovascular reactivity to stress in Black adolescents and young adults.
Cardiovascular reactivity to stress and beta-adrenergic receptor (beta-AR) function may contribute to the development of hypertension. As Black Americans have an increased risk of hypertension, we evaluated associations between beta(1)-AR (Arg389Gly) and beta(2)-AR (Arg16Gly, Gln27Glu) gene variants and cardiovascular reactivity in 500 Black youth. Heart rate, preejection period, total peripheral resistance, and blood pressure reactivity were measured during cold and psychological stress. The Arg389Gly polymorphism in the beta(1)-AR was associated with preejection period reactivity in males but not in females. The Arg16Gly polymorphism in the beta(2)-AR was associated with diastolic blood pressure reactivity only during video game stress. An association between the Gln27Glu polymorphism in the beta(2)-AR and vascular reactivity depended on sex. Thus, specific patterns of associations emerged between genetic variations in beta-ARs and cardiovascular reactivity in young Blacks.


LIBERTY randomized withdrawal study: relugolix combination therapy for heavy menstrual bleeding associated with uterine fibroids.
In the pivotal LIBERTY 1 and 2 trials and long-term extension study, once-daily relugolix combination therapy (40 mg relugolix, 1 mg estradiol, 0.5 mg norethindrone acetate) reduced menstrual blood loss volume and pain among women with uterine fibroids. Relugolix combination therapy was well tolerated with preservation of bone mineral density through 52 weeks. This study aimed to report the 2-year relugolix combination therapy efficacy and safety results of the phase 3 LIBERTY randomized withdrawal study. Women with uterine fibroid-associated heavy menstrual bleeding who completed the 24-week LIBERTY 1 or 2 trials, followed by the 28-week long-term extension study (up to 52 weeks total treatment), and who met the responder criteria (menstrual blood loss volume <80 mL and ≥50% reduction from pivotal study baseline at week 48 [week 24 of long-term extension]) were randomized in a 1:1 ratio to either blinded treatment with relugolix combination therapy or placebo for 52 weeks (total treatment period, 104 weeks). For women who had a relapse of heavy menstrual bleeding during the study (menstrual blood loss volume ≥80 mL), open-label relugolix combination therapy was offered. The primary endpoint was the proportion of women who maintained menstrual blood loss volume <80 mL through week 76 (week 24 of randomized withdrawal study). Secondary endpoints included time to menstrual blood loss volume ≥80 mL, proportion of women who maintained a menstrual blood loss volume of <80 mL through week 104 (over the 52-week randomized treatment period), the proportion of women who achieved or maintained amenorrhea at week 76 at the end of treatment, and the change in Uterine Fibroid Symptom-Quality of Life Bleeding and Pelvic Discomfort Scale and symptom severity scores. Analyses were performed for the modified intent-to-treat population, including all randomized women who received ≥1 dose of the study drug. Of the 229 randomized women (relugolix combination therapy, n=115; placebo, n=114), 228 received the study drug and 175 (76.7%) completed the randomized withdrawal study. Through week 76, 78.4% of women on relugolix combination therapy maintained menstrual blood loss volume <80 mL vs 15.1% in the placebo group (difference, 63.4%; 95% confidence interval, 52.9%-73.9%; P<.0001). At week 104, 69.8% of women on relugolix combination therapy maintained menstrual blood loss volume <80 mL vs 11.8% in the placebo group (difference, 58.0%; 95% confidence interval, 47.0%-69.1%; P<.0001). Through week 104, 88.3% of women on placebo relapsed with heavy menstrual bleeding (median time to relapse, 5.9 weeks). Among the 89 women in the placebo group who relapsed and received open-label rescue treatment, 87 women responded to relugolix combination therapy with a menstrual blood loss volume <80 mL. The proportion of women who achieved or maintained amenorrhea were 57.4% vs 13.3% at week 76 (difference, 44.1%; 95% confidence interval, 33.10%-55.1%; P<.0001) and 58.3% vs 10.6% at week 104 (difference, 47.6%; 95% confidence interval, 37.0%-58.3%; nominal P<.0001) for relugolix combination therapy and the placebo group, respectively. Relugolix combination therapy was generally well tolerated; no new safety signals were identified, and the adverse event profile over the second year was consistent with that reported through the first year of treatment. Bone mineral density remained stable in women who received relugolix combination therapy from week 52 to week 104. In women continuously treated with relugolix combination therapy up to 2 years, bone mineral density was generally preserved. After 2 years of treatment with relugolix combination therapy, there was evidence of durability of the effect in maintaining low menstrual blood loss volume in women with symptomatic uterine fibroids. Most women had return of heavy menstrual bleeding and associated symptoms after treatment cessation, which improved upon retreatment with relugolix combination therapy. Relugolix combination therapy was well tolerated, the adverse event profile remained consistent, and the mean bone mineral density was generally preserved through 2 years of treatment.


Circulating tumor DNA analysis as a real-time method for monitoring tumor burden in melanoma patients undergoing treatment with immune checkpoint blockade.
Assessment of therapeutic activity of drugs blocking immune checkpoints such as CTLA-4 and PD-1/PD-L1 can be challenging, as tumors may seem to enlarge or appear anew before regressing, due to intratumoral inflammation. We assessed whether circulating tumor DNA (ctDNA) levels could serve as an early indicator of true changes in tumor burden in patients undergoing treatment with these agents. Tumors from 12 patients with metastatic melanoma undergoing treatment with checkpoint blocking drugs were analyzed for the presence of hotspot somatic mutations in BRAF, cKIT, NRAS, and TERT. Plasma was collected serially from each patient and levels of ctDNA were compared with radiologic and clinical outcomes. In 5 of 10 patients studied, mutations were detected in BRAF(1), NRAS(2), TERT(1) and ALK(1). Analysis of plasma from 4 of 5 patients identified mutations identical to those found in tumor specimens. Plasma ctDNA levels ranged from undetectable (<0.01%) to 5.5% of total circulating cell-free DNA. In 3 patients, increasing ctDNA levels correlated with progressive disease assessed by radiography. In one patient, ctDNA levels increased after undergoing a needle biopsy of a tumor deposit. In another patient, ctDNA levels increased initially as lymphadenopathy progressed by examination, but then became undetectable 3 weeks prior to clinical improvement. Levels of ctDNA correlated with clinical and radiologic outcomes, and, in one case, preceded eventual tumor regression. Further prospective analysis is required to assess the utility of ctDNA as an early biomarker of clinical outcomes in patients receiving immune checkpoint blocking drugs.


Innovative Medical Technology, Health Technology Assessment, and Health Policy: The Case of Remote Patient Monitoring of Cardiac Implantable Electronic Devices in South Korea.
In many developed countries with universal coverage healthcare systems, payers require new medical technologies to be assessed as safe, effective, and cost-effective through health technology assessment (HTA) before approval for reimbursement coverage and market access. However, in some cases, HTA is not the sole criterion for decision-making and other factors override the evidence. Remote patient monitoring (RPM) for cardiac implantable electronic devices, a novel technology recognized as safe, effective, and cost-effective, and the standard of care in many countries, is prohibited in South Korea. This peculiar situation is apparently due to deficiencies in healthcare policy and the delivery system and also to poor engagement between stakeholders. We propose that a higher level of engagement and trust between stakeholders needs to be developed, and healthcare providers should be involved in the early development of health policy, so that unnecessary barriers to access to useful medical technology are corrected, thereby allowing Koreans to enjoy the benefits available in other developed countries.


[Treatments for chronic sinusitis in children--a study by questionnaire from 433 hospitals in Japan].
Information concerning the medical and surgical treatment of pediatric patients with chronic sinusitis was obtained through questionnaires from 433 hospitals in Japan. The hospitals were authorized as an inservice otorhinolaryngological training institute by the Oto-Rhino Laryngological Society of Japan. The questionnaires were sent to the hospitals in February, 1995, and answers were obtained from 311 hospitals within one month, a 73.5% response rate. Conservative therapies were the most common treatment for pediatric sinusitis. The removal of hypersecreted nasal mucus by aspiration in combination with alphaadrenergic decongestants was performed in 89.1% of the hospitals. Treatment by oral administration of drugs was adapted in 88.1%. The administration of antibiotics and/or steroids by aerosol was performed in 86.2%. The antral lavage was performed in only 17.7%. In the medical treatment, macrolide antibiotics and proteolytics enzymes were most frequently administered. Surgical therapy was performed if the patient was indicated for surgery in 241 hospitals (77.5%). Nasal polyps were removed surgically in 202 hospitals. The intranasal ethmoidectomy was performed in 98 hospitals. Five years of age was the youngest limit of the surgical indication. In most hospitals, the population of surgically treated patients was less than 5% of all pediatric patients with chronic sinusitis. Adenoidectomy was adapted for the improvement of chronic sinusitis in 33.8% of the hospitals.


Genetic data reveal mixed-stock aggregations of gray whales in the North Pacific Ocean.
Gray whales (Eschrichtius robustus) in the Western Pacific are critically endangered, whereas in the Eastern Pacific, they are relatively common. Holocene environmental changes and commercial whaling reduced their numbers, but gray whales in the Eastern Pacific now outnumber their Western counterparts by more than 100-fold. Herein, we investigate the genetic diversity and population structure within the species using a panel of genic single nucleotide polymorphisms. Results indicate the gray whale gene pool is differentiated into two substocks containing similar levels of genetic diversity, and that both our Eastern and Western geographical samples represent mixed-stock aggregations. Ongoing or future gene flow between the stocks may conserve genetic diversity overall, but admixture has implications for conservation of the critically endangered Western gray whale.


Tiglylglycine excreted in urine in disorders of isoleucine metabolism and the respiratory chain measured by stable isotope dilution GC-MS.
Tiglyglycine (TG), an intermediate product of the catabolism of isoleucine, is increased in the urine of patients with beta-ketothiolase deficiency or with disorders of propionate metabolism. It is also implicated as a useful diagnostic marker in disorders of the respiratory chain. We present a method for the synthesis of TG and tiglyl[13C, 15N]glycine and the development of a stable isotope dilution mass spectrometric assay for TG. We compare data from controls with that from subjects with beta-ketothiolase deficiency and propionyl-CoA carboxylase deficiency, and with six patients with enzyme-confirmed disorders of the respiratory chain. TG was increased in the urine from all of the patient groups. The increased TG excretion did not persist in one patient with a respiratory chain defect, which suggests that, in some patients, multiple sample analysis may be necessary to identify a respiratory chain defect. This is the first urinary compound to be implicated as a potential marker of disorders of the respiratory chain.


Revisiting the influence of chain length on the α- and β-relaxations in oligomeric glass formers.
Dielectric relaxation measurements of a series of oligo(propylene glycol) dimethyl ethers, CH3-O-[CH2-CH(CH3)-O]N-CH3, including samples with the number of PG units N = 1, 2, 3, 7, 17, 34, and 69, were made by Mattsson et al. [Phys. Rev. Lett. 94, 165701 (2005)] at ambient pressure. The objective of the study was to relate the change of properties of the glass transition dynamics to the number of monomer units N in the chain. Not examined in the previous publication is how the change of the width of the frequency dispersion of the α-relaxation with N is related to the observed change in the α-β bifurcation characterized by the ratio, τ(α)(T(g))/τ(β)(T(g)). In this paper, the frequency dispersion of the dimer, trimer, and heptamer are fitted by the Fourier transform of the Kohlrausch stretched exponential function, φ(t) = exp[-(t/τ(α))(1-n)]. Determined from experimental data, both τ(α)(T(g))/τ(β)(T(g)) and n increase with N. More interestingly, we find τ(α)(T(g))/τ(β)(T(g)) has approximately the same value as [τ(α)(T(g))/t(c)](n) with t(c) = 2 ps, in accordance with the prediction of the Coupling Model of approximate relation between τ(α) and τ(β) given by τ(β) ≈ (t(c))(n)(τ(α))(1-n). Considered also are previously unpublished dielectric loss spectra of the heptamer taken at different combinations of T and P with τ(α)(T,P) fixed by Roland et al. [Phys. Rev. B 77, 012201 (2008)]. The dielectric loss data show not only the α-loss peaks superpose but also the high frequency flank including the barely resolved JG β-relaxation superposes approximately. This is again consistent with the approximate relation between τ(α) and τ(β) from the Coupling Model because n is unchanged on varying P and T with τ(α)(T,P) kept constant, and t(c) is a constant. The additional advance made herein has the benefit of enhancing the impact of the earlier experimental studies of the oligo(propylene glycol) dimethyl ethers on current understanding of the dynamics of glass transition.


Evaluation of spectral domain optical coherence tomography parameters in discriminating preperimetric glaucoma from high myopia.
To evaluate the diagnostic ability of macular ganglion cell-inner plexiform layer (GCIPL) thickness obtained by spectral-domain optical coherence tomography (SD-OCT) in discriminating non-highly myopic eyes with preperimetric glaucoma (PPG) from highly myopic healthy eyes. A total of 254 eyes, including 76 normal controls (NC), 116 eyes with high myopia (HM) and 62 non-highly myopic eyes with PPG were enrolled. The diagnostic ability of OCT parameters was accessed by the areas under the receiver operating characteristic (AUROC) curve in two distinguishing groups: PPG eyes with non-glaucomatous eyes including NC and HM (Group 1), and PPG eyes with HM eyes (Group 2). Differences in diagnostic performance between GCIPL and RNFL parameters were evaluated. The minimum (AUROC curve of 0.782), inferotemporal (0.758) and inferior (0.705) GCIPL thickness were the top three GCIPL parameters in discriminating PPG from non-glaucomatous eyes, all of which had statistically significant lower diagnostic ability than average RNFL thickness (0.847). In discriminating PPG from HM, the best GCIPL parameter was minimum (0.689), statistically significant lower in diagnostic ability than average RNFL thickness (0.789) and three other RNFL thickness parameters of temporal and inferotemporal clock-hour sectors. The minimum GCIPL thickness is the best GCIPL parameter to detect non-highly myopic PPG from highly myopic eyes, whose diagnostic ability is inferior to that of average RNFL thickness and RNFL thickness of several temporal and inferotemporal clock-hour sectors. The average RNFL thickness is recommended for discriminating PPG from highly myopic healthy eyes in current clinical practice in a Chinese population.


[Social Interaction: The experiences of COVID-19 positive women with perinatal risk].
The COVID-19 pandemic induced changes in the dynamics of the life of women in the perinatal phase who, due to the health crisis, restructured social care and coexistence practices. The scope of this paper was to assess the experiences of high-risk perinatal pregnancy risk among COVID-19 positive women through social interaction. The work was conducted in a hospital of tertiary perinatal care. Qualitative methodology was used, whereby questionnaires and interviews were conducted via zoom with 14 COVID-19 positive women in the perinatal phase. Critical-interpretative discourse analysis was applied based on the concept of social interaction and complex thinking. Three types of social interaction were developed to assess the results: a) Initial social interaction: experiences when becoming aware of being COVID-19 positive; b) Acquired social interaction: experiences of care prior to COVID-19; c) Enduring social interaction: experience required in the face of COVID-19. The result of experience leads to new forms of social interaction after notification ranging from care to resilience. The conclusion drawn is that the experience of COVID-19 of women in the perinatal period remodeled their ways of coexistence and care within the institutional, family, and personal spheres.


Selective Adsorption and Separation of Organic Dyes with Spherical Polyelectrolyte Brushes and Compressed Carbon Dioxide.
Dye-containing wastewater has caused serious environmental pollution. Herein, rationally designed spherical polyelectrolyte brushes (SPBs) with cationic charges, polystyrene-poly(2-aminoethylmethacrylate hydrochloride) (PS-PAEMH) as the absorbent, and compressed carbon dioxide as the antisolvent are proposed for the separation of the anionic dye eosin Y (EY) from a solution of mixed dyes. The adsorption behavior of EY onto PS-PAEMH was highly dependent on CO2 pressure, contact time, and initial concentration. The maximum adsorption capacity of PS-PAEMH was 335.20 mg g-1 . FTIR and UV/Vis measurements proved that the electrostatic interactions between EY and PS-PAEMH played an important role in the absorbance process. The adsorption process fitted the pseudo-second-order kinetic model and Freundlich isotherm model very well. The combined dye and polymer brush could be easily separated through ion exchange by adding an aqueous solution of NaCl. Recovered PS-PAEMH retained a high adsorption capacity even after ten cycles of regeneration. This method provides a simple and effective way to separate ionic materials for environmental engineering.


Identification and expression in mouse of two heparan sulfate glucosaminyl N-deacetylase/N-sulfotransferase genes.
The biosynthesis of heparan sulfate/heparin is a complex process that requires the coordinate action of a number of different enzymes. In close connection with polymerization of the polysaccharide chain, the modification reactions are initiated by N-deacetylation followed by N-sulfation of N-acetylglucosamine units. These two reactions are carried out by a single protein. Proteins with such dual activities were first purified and cloned from rat liver and mouse mastocytoma. The mouse mastocytoma enzyme is encoded by an approximately 4-kilobase (kb) mRNA, whereas the rat liver transcript contains approximately 8 kb. In the present study, the primary structure of the enzyme encoded by the mouse 8-kb transcript is described. It is demonstrated that both the 4-and 8-kb transcripts have a wide tissue distribution and that they are encoded by separate genes. Characterization of the gene encoding the 4-kb transcript demonstrates that it spans a region of about 8 kb and that it contains at least 14 exons. The similarity of this gene and the previously characterized human gene for the 8-kb transcript is discussed.


A 9-14 year follow-up of onlay bone grafting in the atrophic maxilla.
Treatment of the atrophic edentulous maxilla is challenging especially when bone graft procedures are necessary. In this study an onlay bone graft, a saddle or veneer, with or without maxillary sinus floor inlay graft, harvested from the anterior iliac crest, in combination with implants was used in the reconstruction of patients with extreme atrophy in their maxillae. The aim was to investigate treatment outcome, and the impact of gender and smoking, in 44 patients in a prospective, long-term, follow-up study concerning implant survival rate and marginal bone loss adjacent to the surfaces of the implant. Mean follow-up time was 11 years. Of 334 inserted Brånemark implants, with machined surface, 27 failed. Estimated implant survival rate was 90%. Marginal bone loss was 1.8 mm 1 year after implant surgery; 2.3 mm after 5 years; and 2.4 mm after 10 years. There was a significant difference between genders in implant survival. Marginal bone loss differed significantly between smokers and non-smokers up to the 5-year examination and between genders after the 4-year examination. The onlay bone graft, with or without a maxillary inlay graft, results in high implant survival rate, good oral function and stabilised marginal bone. All patients are still wearing their original fixed bridges.


Ligands Exert Biased Activity to Regulate Sigma 1 Receptor Interactions With Cationic TRPA1, TRPV1, and TRPM8 Channels.
The sigma 1 receptor (σ1R) and the mu-opioid receptor (MOR) regulate the transient receptor potential (TRP) V1 calcium channel. A series of proteins are involved in the cross-regulation between MORs and calcium channels like the glutamate N-methyl-D-aspartate receptor (NMDAR), including the histidine triad nucleotide-binding protein 1 (HINT1), calmodulin (CaM), and the σ1R. Thus, we assessed whether similar mechanisms also apply to the neural TRP ankyrin member 1 (TRPA1), TRP vanilloid member 1 (TRPV1), and TRP melastatin member 8 (TRPM8). Our results indicate that σ1R and CaM bound directly to cytosolic regions of these TRPs, and this binding increased in the presence of calcium. By contrast, the association of HINT1 with these TRPs was moderately dependent on calcium. The σ1R always competed with CaM for binding to the TRPs, except for its binding to the TRPA1 C-terminal where σ1R binding cooperated with that of CaM. However, σ1R dampened HINT1 binding to the TRPA1 N-terminal. When the effect of σ1R ligands was addressed, the σ1R agonists PRE084 and pregnenolone sulfate enhanced the association of the σ1R with the TRPM8 N-terminal and TRPV1 C-terminal in the presence of physiological calcium, as seen for the σ1R-NMDAR interactions. However, these agonists dampened σ1R binding to the TRPA1 and TRPV1 N-terminal domains, and also to the TRPA1 C-terminal, as seen for σ1R-binding immunoglobulin protein (BiP) interactions in the endoplasmic reticulum (ER). By contrast, the σ1R antagonists progesterone and S1RA reduced the association of σ1R with TRPA1 and TRPV1 C-terminal regions, as seen for the σ1R-NMDAR interactions. Conversely, they enhanced the σ1R interaction with the TRPA1 N-terminal, as seen for σ1R-BiP interactions, whereas they barely affected the association of σ1R with the TRPV1 N-terminal. Thus, depending on the calcium channel and the cytosolic region examined, the σ1R agonists pregnenolone sulfate and PRE084 opposed or collaborated with the σ1R antagonists progesterone and S1RA to disrupt or promote such interactions. Through the use of cloned cytosolic regions of selected TRP calcium channels, we were able to demonstrate that σ1R ligands exhibit biased activity to regulate particular σ1R interactions with other proteins. Since σ1Rs are implicated in essential physiological processes, exploiting such ligand biases may represent a means to develop more selective and efficacious pharmacological interventions.


Validity of classical nucleation theory for Ising models.
While the classical nucleation theory (CNT) is widely used to predict the rate of first-order phase transitions, its validity has been questioned due to discrepancies with experiments. We systematically test the individual components of CNT by computer simulations of the Ising models and confirm its fundamental assumptions under a wide range of conditions ( h=0.01-0.13J , T=0.44-0.84Tc in two-dimensions and h=0.30-0.60J , T=0.48-0.62Tc in three dimensions). First, CNT accurately predicts the nucleation rate if the correct droplet free energy is provided. Furthermore, theoretical prediction of droplet free energy matches numerical results very well in the two-dimensional (2D) Ising model, if appropriate correction terms are added. This establishes the 2D Ising model as an important reference point where existing theories can predict nucleation rate accurately with no adjustable parameters.


Lower urinary tract symptoms among Caucasian-European men who have sex with men: findings from a real-life survey.
Prevalence of and severity of lower urinary tract symptoms (LUTS) according to male sexual orientation have been scantly analysed. We aimed to assess the prevalence and severity of LUTS in a cohort of Caucasian-European men who have sex with men seeking medical help for uroandrologic reasons other than LUTS. Data from 949 consecutive individuals in an outpatient setting were analysed. Severity of LUTS was measured with the International Prostate Symptom Score (IPSS). Men with storage symptoms scored 1-3 and ⩾ 4 (of 15), and voiding symptoms scored 1-4 and ⩾ 5 (of 20) were considered as having mild and moderate-to-severe symptoms, respectively. For individual symptoms, patients with scores ⩾ 1 were deemed symptomatic (according to Apostolidis et al.(15)). Descriptive statistics and logistic regression models tested the association between LUTS and sexual orientation. Complete data were available for 213 (22.4%) men who have sex with men (MSM) and 736 (77.6%) heterosexuals (mean age (s.d.): 41.0 (12.2) vs 39.9 (12.1) years). Compared with heterosexuals, MSM reported higher rates of total IPSS scores suggestive of moderate (21.6% vs 20%) and severe LUTS (3.8% vs 2.4%) (P=0.004). Similarly, MSM showed higher rates of mild (48.8% vs 45.2%) and moderate-to-severe (39.4% vs 30.4%) storage symptoms (all P<0.001), and of mild (45.1% vs 34.8%) and moderate-to-severe (20.2% vs 19.2%) voiding symptoms (all P<0.01). MSM status was an independent predictor of mild voiding symptoms (odds ratio (OR): 1.40; P=0.004), moderate-to-severe storage symptoms (OR: 1.40; P=0.04) and severe total IPSS (OR: 1.49; P=0.03), after adjusting for other variables. These findings suggest a higher prevalence and severity of LUTS in MSM compared with heterosexual men seeking medical help for uroandrologic reasons other than LUTS.


Epizootiology, pathology, and microbiology of an outbreak of urolithiasis in chickens.
An outbreak of urolithiasis that doubled the annual mortality rate of chickens in a large flock of table-egg-layers is described. Despite the presence of a large unilateral urolith and/or severe renal atrophy, the layers often maintained active egg production and apparent homeostasis until a small urolith blocked the ureteral flow from the contralateral kidney. This terminal episode appeared to produce acute obstructive renal failure, rapidly developing visceral gout (visceral urate deposition), uremia, and death. The atrophy observed appeared to be acquired and progressive. Histologic features in the kidneys were acute to chronic glomerulonephritis, interstitial nephritis, and pyelonephritis. Epizootiologic and microbiologic studies indicated that a combination of infectious and noninfectious mechanisms may have been involved. Causative roles for calcium-phosphate imbalance, infectious bronchitis (IB), Newcastle disease (ND), and adenovirus or reovirus infections could be neither excluded nor confirmed. Contributory factors may have been spray ND-IB and other vaccinations of 15-week-old ND-IB-susceptible pullets, water deprivation, shipping stress, Mycoplasma synoviae infection, immune complex disease, and mycotoxins.


Sjögren-Larsson syndrome: definitive diagnosis on magnetic resonance spectroscopy.
Sjögren-Larsson syndrome (SLS) is a rare autosomal-recessive neurocutaneous disorder comprising a triad of ichthyosis, mental retardation, and spastic diplegia or quadriplegia. It has rarely been reported in Asian and Indian populations. We report the case of an Indian patient with SLS who presented with the classical clinical triad and demonstrated characteristic findings on magnetic resonance spectroscopy. In resource-restricted settings where enzymatic and genetic analyses are not available, magnetic resonance spectroscopy serves as a useful adjunct in confirming the diagnosis of SLS.


Neuronal deficiency of ARV1 causes an autosomal recessive epileptic encephalopathy.
We report an individual who presented with severe neurodevelopmental delay and an intractable infantile-onset seizure disorder. Exome sequencing identified a homozygous single nucleotide change that abolishes a splice donor site in the ARV1 gene (c.294 + 1G > A homozygous). This variant completely prevented splicing in minigene assays, and resulted in exon skipping and an in-frame deletion of 40 amino acids in primary human fibroblasts (NP_073623.1: p.(Lys59_Asn98del). The p.(Lys59_Asn98del) and previously reported p.(Gly189Arg) ARV1 variants were evaluated for protein expression and function. The p.(Gly189Arg) variant partially rescued the temperature-dependent growth defect in arv1Δ yeast, while p.(Lys59-Asn98del) completely failed to rescue at restrictive temperature. In contrast to wild type human ARV1, neither variant expressed detectable levels of protein in mammalian cells. Mice with a neuronal deletion of Arv1 recapitulated the human phenotype, exhibiting seizures and a severe survival defect in adulthood. Our data support ARV1 deficiency as a cause of autosomal recessive epileptic encephalopathy.


PRMT5 Inhibitors Regulate DNA Damage Repair Pathways in Cancer Cells and Improve Response to PARP Inhibition and Chemotherapies.
Expression of protein arginine methyltransferase 5 (PRMT5) is highly positively correlated to DNA damage repair (DDR) and DNA replication pathway genes in many types of cancer cells, including ovarian and breast cancer. In the current study, we investigated whether pharmacologic inhibition of PRMT5 downregulates DDR/DNA replication pathway genes and sensitizes cancer cells to chemotherapy and PARP inhibition. Potent and selective PRMT5 inhibitors significantly downregulate expression of multiple DDR and DNA replication genes in cancer cells. Mechanistically, PRMT5 inhibition reduces the presence of PRMT5 and H4R3me2s on promoter regions of DDR genes such as BRCA1/2, RAD51, and ATM. PRMT5 inhibition also promotes global alternative splicing changes. Our data suggest that PRMT5 inhibition regulates expression of FANCA, PNKP, and ATM by promoting exon skipping and intron retention. Combining C220 or PRT543 with olaparib or chemotherapeutic agents such as cisplatin demonstrates a potent synergistic interaction in breast and ovarian cancer cells in vitro. Moreover, combination of PRT543 with olaparib effectively inhibits the growth of patient-derived breast and ovarian cancer xenografts. Furthermore, PRT543 treatment significantly inhibits growth of olaparib-resistant tumors in vivo. These studies reveal a novel mechanism of PRMT5 inhibition and suggest beneficial combinatorial effects with other therapies, particularly in patients with tumors that are resistant to therapies dependent on DNA damage as their mechanism of action. Patients with advanced cancers frequently develop resistance to chemotherapy or PARP inhibitors mainly due to circumvention and/or restoration of the inactivated DDR pathway genes. We demonstrate that inhibition of PRMT5 significantly downregulates a broad range of the DDR and DNA replication pathway genes. PRMT5 inhibitors combined with chemotherapy or PARP inhibitors demonstrate synergistic suppression of cancer cell proliferation and growth in breast and ovarian tumor models, including PARP inhibitor-resistant tumors.


Intraosseous neurilemmoma of the cervical spine causing paraparesis and treated by resection and grafting.
A neurilemmoma presented as an expanding lesion of the bodies of C6 and C7 vertebrae and caused local neurological signs together with a spastic paraparesis. This tumour was treated by preliminary posterior fusion, followed by its complete removal via an anterior approach and stabilization by anterior spinal fusion. Other cases in the literature are reviewed and discussed.