Cytochrome b phylogeography of chamois (Rupicapra spp.). Population contractions, expansions and hybridizations governed the diversification of the genus.
The chamois provides an excellent model for exploring the effect of historical and evolutionary events on diversification. We investigate cytochrome b (cytb) sequences in the 10 recognized subspecies of Rupicapra classified within 2 species: Rupicapra pyrenaica, with the subspecies parva, pyrenaica, and ornata, and Rupicapra rupicapra, with cartusiana, rupicapra, tatrica, carpatica, balcanica, asiatica, and caucasica. A fragment of 349 bp of the cytb was sequenced in 189 individuals. We identified 3 cytb lineages: Clade West in Iberia and Western Alps; Clade Central in the Apennines and the Massif of Chartreuse; and Clade East present in populations to the east of the Alps. The 2 proposed species were polyphyletic; the clades West and Central are represented in both, whereas the Clade East is restricted to R. rupicapra. In contrast to the current systematic, cytb phylogenies suggest the classification of the 10 subspecies of chamois into a single species, R. rupicapra. Phylogeny and geographical distribution of the 3 lineages show the effects of limited latitudinal range expansions, contractions, and hybridizations among highly divergent lineages, along with a major role of the glacial ice sheets of the Alps and the Pyrenees as barriers to gene flow, on the diversification of extant taxa.


Safety of iron hydroxide adipate tartrate as a novel food pursuant to Regulation (EU) 2015/2283 and as a source of iron in the context of Directive 2002/46/EC.
Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on iron hydroxide adipate tartrate as a novel food (NF) pursuant to Regulation (EU) 2015/2283 and as a source of iron in the context of Directive 2002/46/EC. The NF is intended to be used in food supplements up to a maximum dose of 100 mg per day, corresponding to a maximum daily intake of iron of 36 mg. The target population proposed by the applicant is the general population above 3 years of age. The NF which is the subject of the application is an engineered nanomaterial having primary particles, of almost spherical morphology, with a diameter typically smaller than 5 nm. The studies provided for absorption, distribution, metabolism and excretion (ADME) and bioavailability indicate that iron, once taken up into the epithelial cells of the gut, is subject to the same mechanisms of regulation and absorption as that of other forms of iron. Further studies provided in the context of the toxicological assessment indicate that the NF does not lead to iron bioaccumulation in tissues and organs at the doses tested. The Panel notes that the NF contains nickel at concentrations that may increase the risk of flare-up reactions in nickel-sensitised young individuals up to 10 years of age. In the 90-day toxicity study, findings related to haematology, clinical biochemistry and organ weights were observed and the Panel defined a no observed adverse effect level (NOAEL) of 231 mg/kg body weight (bw) per day, that is, the mid-dose used in the study. The Panel considers that the NF is a source from which iron is bioavailable and it is safe under the proposed conditions of use.


Assessing reliable human mobility patterns from higher order memory in mobile communications.
Understanding how people move within a geographical area, e.g. a city, a country or the whole world, is fundamental in several applications, from predicting the spatio-temporal evolution of an epidemic to inferring migration patterns. Mobile phone records provide an excellent proxy of human mobility, showing that movements exhibit a high level of memory. However, the precise role of memory in widely adopted proxies of mobility, as mobile phone records, is unknown. Here we use 560 million call detail records from Senegal to show that standard Markovian approaches, including higher order ones, fail in capturing real mobility patterns and introduce spurious movements never observed in reality. We introduce an adaptive memory-driven approach to overcome such issues. At variance with Markovian models, it is able to realistically model conditional waiting times, i.e. the probability to stay in a specific area depending on individuals' historical movements. Our results demonstrate that in standard mobility models the individuals tend to diffuse faster than observed in reality, whereas the predictions of the adaptive memory approach significantly agree with observations. We show that, as a consequence, the incidence and the geographical spread of a disease could be inadequately estimated when standard approaches are used, with crucial implications on resources deployment and policy-making during an epidemic outbreak.


Thyroid autoimmunity and Behçet's disease: is there a significant association?
Behcet's disease (BD) could be regarded as an autoimmune disease in many aspects. Autoimmune thyroid disease (ATD) is frequently accompanied by other various autoimmune diseases. Nevertheless, there is not still enough data showing the association between BD and ATD. In addition, no controlled study is present in the PubMed, which evaluates thyroidal autoimmunity using antithyroid peroxidase antibody in a large series of patients with BD. We aimed to investigate the frequency of ATD in patients with BD. The study included 124 patients with BD and 99 age- and sex-matched healthy volunteers. Autoimmune thyroiditis was noted in 21 cases (16.9%) with BD. In the control group, 22 cases (22.22%) were diagnosed as autoimmune thyroiditis. There was no difference between the groups in respect to thyroid autoantibodies (P>0.05). There were no statistically significant differences between baseline TSH levels of the BD patients and of the controls (P>0.05). Statistically, the mean serum free T4 levels of the patients with BD were higher than those of the controls (P<0.001). No association could be found between BD and ATD. Therefore, it is not of significance to investigate thyroid autoimmunity in BD.


Fine particulate matter source apportionment for the Chemical speciation Trends Network site at Birmingham, Alabama, using Positive Matrix Factorization.
The Positive Matrix Factorization (PMF) receptor model version 1.1 was used with data from the fine particulate matter (PM2.5) Chemical Speciation Trends Network (STN) to estimate source contributions to ambient PM2.5 in a highly industrialized urban setting in the southeastern United States. Model results consistently resolved 10 factors that are interpreted as two secondary, five industrial, one motor vehicle, one road dust, and one biomass burning sources. The STN dataset is generally not corrected for field blank levels, which are significant in the case of organic carbon (OC). Estimation of primary OC using the elemental carbon (EC) tracer method applied on a seasonal basis significantly improved the model's performance. Uniform increase of input data uncertainty and exclusion of a few outlier samples (associated with high potassium) further improved the model results. However, it was found that most PMF factors did not cleanly represent single source types and instead are "contaminated" by other sources, a situation that might be improved by controlling rotational ambiguity within the model. Secondary particulate matter formed by atmospheric processes, such as sulfate and secondary OC, contribute the majority of ambient PM2.5 and exhibit strong seasonality (37 +/- 10% winter vs. 55 +/- 16% summer average). Motor vehicle emissions constitute the biggest primary PM2.5 mass contribution with almost 25 +/- 2% long-term average and winter maximum of 29 +/- 11%. PM2.5 contributions from the five identified industrial sources vary little with season and average 14 +/- 1.3%. In summary, this study demonstrates the utility of the EC tracer method to effectively blank-correct the OC concentrations in the STN dataset. In addition, examination of the effect of input uncertainty estimates on model results indicates that the estimated uncertainties currently being provided with the STN data may be somewhat lower than the levels needed for optimum modeling results.


[Surgical treatment of supraventricular tachycardia].
Surgical treatment of supraventricular tachycardia was performed in 61 patient, including the 52--with additional atrio-ventricular connections. Arrhythmia was liquidated in 57 (93.4%) patients. Hospital lethality was 1.7%.


[Childbirth-related fatigue during labor: an important but overlooked symptom].
Fatigue is not only a distressful symptom that is prevalent during labor; it may also interfere with the ability of laboring women to adjust to pain and bear down in the second labor stage as well as hinder maternal recovery and impair postpartum maternal-role performance. Childbirth-related fatigue is a frequent cause of physical and mental discomfort during labor and increases the likelihood of medical interventions such as instrumental delivery and cesarean section. Even so, as this fatigue is widely regarded as an inevitable part of the labor experience, little attention has been given to mediative interventions. In light of the above, this paper illustrates the definition and characteristics of childbirth-related fatigue during labor, analyzes the factors that affect this fatigue, and summarizes relevant nursing-care principles. It is hoped this paper increases nurse awareness of childbirth-related fatigue during labor and helps nurses take appropriate early prevention and intervention measures to reduce the adverse effects of fatigue and improve intrapartum care quality.


Activity-based profiling of a physiologic aglycone library reveals sugar acceptor promiscuity of family 1 UDP-glucosyltransferases from grape.
Monoterpenols serve various biological functions and accumulate in grape (Vitis vinifera), where a major fraction occurs as nonvolatile glycosides. We have screened the grape genome for sequences with similarity to terpene URIDINE DIPHOSPHATE GLYCOSYLTRANSFERASES (UGTs) from Arabidopsis (Arabidopsis thaliana). A ripening-related expression pattern was shown for three candidates by spatial and temporal expression analyses in five grape cultivars. Transcript accumulation correlated with the production of monoterpenyl β-d-glucosides in grape exocarp during ripening and was low in vegetative tissue. Targeted functional screening of the recombinant UGTs for their biological substrates was performed by activity-based metabolite profiling (ABMP) employing a physiologic library of aglycones built from glycosides isolated from grape. This approach led to the identification of two UDP-glucose:monoterpenol β-d-glucosyltransferases. Whereas VvGT14a glucosylated geraniol, R,S-citronellol, and nerol with similar efficiency, the three allelic forms VvGT15a, VvGT15b, and VvGT15c preferred geraniol over nerol. Kinetic resolution of R,S-citronellol and R,S-linalool was shown for VvGT15a and VvGT14a, respectively. ABMP revealed geraniol as the major biological substrate but also disclosed that these UGTs may add to the production of further glycoconjugates in planta. ABMP of aglycone libraries provides a versatile tool to uncover novel biologically relevant substrates of small-molecule glycosyltransferases that often show broad sugar acceptor promiscuity.


The retrotransposon copia regulates Drosophila gene expression both positively and negatively.
The D. melanogaster transposable element copia is structurally similar to retroviral proviruses. We have asked whether copia encodes regulatory functions which have been observed in certain other proviruses. We have introduced reporter constructs based on the copia promoter and other Drosophila promoters into Drosophila cells and asked if copia has any affect on their expression. We find that, whilst copia negatively regulates expression from its own promoter, it also positively regulates expression from the larval serum protein 1 promoter. Analysis of RNA suggests that both regulatory functions occur by post-transcriptional mechanisms.


[2008 G8 Hokkaido Toyako Summit Meeting Syndrome Surveillance].
We conducted syndromic surveillance for the Hokkaido, Japan, Toyako Group of Eight (G8) summit meeting in July 2008 as a counter-measure to bioterrorism attacks and other health emergencies. Surveys were conducted from June 23, two weeks before the summit, to July 23 two weeks after it, with part of those for prescription drugs fully automated, and part by manual input over the World-Wide-Web. Those for ambulance transfer were done similarly. We bought over-the-counter (OTC) sales data from two private research firms in Japan and had the monitor, who had contacts with a private research company, report health conditions via personal computer (PC) or cellphone. We had a virtual conference daily at 9:00 with the local Hokkaido government, local public health center, local Hokkaido public laboratory, the National Institute of Infectious Diseases, and the Ministry of Health, Labor and Welfare to decide whether local public health centers would be required to investigate. Fully automated syndromic surveillance was conducted by 23 pharmacies for prescriptions drugs, and 71 pharmacies provided manual corporate input. One fire department covering Toyako and a VIP support team used fully automated syndromic surveillance and seven Toyako fire departments used manual input. For 79 pharmacies providing OTC sales data, data provision was delayed one day and analysis could not be automated. Four hundred and seventy two households corporate web search for their health conditions. It also automatically analyzed and feed backed. No notable outbreak occurred during the summit, but public health centers investigated seven aberration detected by syndrome surveillance for ambulance transfer. Although a fully automated system was concidered best for early outbreak detection manual input and analysis were also required. Routine, fully automatied syndromic surveillance remains to be realized in Japan.


Thymoquinone and geraniol alleviate cisplatin-induced neurotoxicity in rats through downregulating the p38 MAPK/STAT-1 pathway and oxidative stress.
Cisplatin (CP) is a widely used broad-spectrum antineoplastic agent used to treat a variety of human malignancies. Neurotoxicity is clinically evident in patients who have undergone a full course of chemotherapy. The aim of this study was to investigate the possible protective effects of thymoquinone (TQ) and geraniol (Ger) against CP-induced neurotoxicity in rats. Forty male Wistar albino rats were allocated into four groups as follows: normal control, CP-induced neurotoxicity, CP + TQ and CP + Ger. Our results demonstrated that simultaneous treatment with either TQ or Ger and CP significantly abrogated oxidative stress and downregulated the apoptotic markers p38 mitogen-activated protein kinase (MAPK), STAT-1, p53, p21 and MMP9; FMO3, however, was insignificantly decreased. In addition to the biochemical results, we assessed the histopathological findings, which confirmed the protective effect of TQ and Ger against the brain damage induced by CP. The results of the present study indicate that simultaneous treatment with either TQ or Ger as natural antioxidants can provide protection against cisplatin-induced neurotoxicity in rats by attenuating oxidative stress and cell apoptosis.


Anti-CD154 (CD40L) prevents recurrence of diabetes in islet isografts in the DR-BB rat.
Islet transplantation for the treatment of autoimmune diabetes is more difficult because of the additional barrier presented by the autoimmunity. We tested the ability of hamster anti-rat CD154 to prevent recurrence of diabetes in renal subcapsular islet isografts in DR-BB (RT1uu) rats with established autoimmune diabetes. Experimental animals with established diabetes received intravenous injections of 15 mg/kg anti-CD154 on a specified schedule starting 2 days before renal subcapsular transplantation of an islet isograft. Control animals received either saline or hamster IgG. Plasma glucose levels >250 mg/dl over 3 days were used to indicate the recurrence of diabetes. Rats that received saline (n = 5) or control antibody (n = 3) had a recurrence of diabetes 6-11 days after transplantation. Histological examination of islet isografts from these rats showed complete destruction of the insulin-producing portion of the isograft with residual cells positive for glucagon. Recipient rats that received anti-CD154 at the 15-mg/kg dosage (n = 6) did not have a recurrence of diabetes for 308-461 days after transplantation. Islet isografts removed from the rats showed low levels of insulin immunoreactivity, high levels of insulin mRNA, and focal infiltration with lymphocytes but no evidence of islet destruction. Mean peak antibody concentration was 266 microg/ml and returned to undetectable levels by 67-88 days after transplantation. Rats that received anti-CD154 starting at 4-7 days after transplantation had a recurrence of diabetes within 11 days of the isotransplantation. Therefore, anti-CD154 as the sole immunomodulator prevented the recurrence of diabetes in islet isografts in rats with established autoimmune diabetes. This suggests that CD40/CD154 blockade is effective in preventing the insulitis or the effector phase of autoimmune diabetes.


Mutated human beta3-adrenergic receptor (Trp64Arg) lowers the response to beta3-adrenergic agonists in transfected 3T3-L1 preadipocytes.
Wild-type or mutated human beta3-adrenergic receptor (Trp64Arg) cDNAs were stably expressed in mouse 3T3-L1 cells. Saturation binding study using a beta-adrenergic ligand revealed that there was no significant difference in the receptor density and the equilibrium dissociation constant between the two cell lines. However, the ability of the mutant beta3-adrenergic receptor to accumulate cyclic AMP (cAMP) in response to isoproterenol was much reduced and Kact for cAMP accumulation was lowered as compared to the wild type receptor. The amount of alpha subunit of stimulatory GTP-binding protein (GSalpha) and adenylyl cyclase activity in response to forskolin were not different in the two cell lines. The responses of the mutant receptor to epinephrine, norepinephrine and L-755,507, a highly specific agonist for human beta3-adrenergic receptor, were also reduced, but the reduction of Kact for L-755,507 was more evident than other agonists tested. The cAMP accumulation in response to some conventional beta3 agonists was less than 10% of that to isoproterenol even in the cells expressing the wild type receptor. These results suggest that the Trp64Arg mutant beta3-adrenergic receptor has less ability to stimulate adenylyl cyclase, and that lipolytic activity through the beta3-adrenergic receptor by catecholamines in subjects carrying this mutation might be suppressed.


Generation of volume data by episcopic three-dimensional imaging of embryos.
Episcopic fluorescence image capturing (EFIC) and high-resolution episcopic microscopy (HREM) are related techniques that are used to generate digital volume data and create three-dimensional (3D) images. Both techniques require specimens that are embedded in an appropriate medium, and images are captured from successive sections before removal from the embedded tissue block. EFIC detects autofluorescence emitted from the embedded tissue, whereas HREM requires the tissue to be stained with a fluorescent dye such as eosin. Volume data are generated as the successive sections are imaged and removed using a microtome. This protocol describes the procedure for digital data processing, visualization, and archiving for both EFIC and HREM.


Repeat prostate biopsy accuracy: simulator-based comparison of two- and three-dimensional transrectal US modalities.
To compare the accuracy of biopsy with two-dimensional (2D) transrectal ultrasonography (US) with that of biopsy with conventional three-dimensional (3D) transrectal US and biopsy with guided 3D transrectal US in the guidance of repeat prostate biopsy procedures in a prostate biopsy simulator. The institutional review board approved this retrospective study. Five residents and five experts performed repeat biopsies with a biopsy simulator that contained the transrectal US prostate images of 10 patients who had undergone biopsy. Simulated repeat biopsies were performed with 2D transrectal US, conventional 3D transrectal US, and guided 3D transrectal US (an extension of 3D transrectal US that enables active display of biopsy targets). The modalities were compared on the basis of time per biopsy and how accurately simulated repeat biopsies could be guided to specific targets. The probability for successful biopsy of a repeat target was calculated for each modality. Guided 3D transrectal US was significantly (P < .01) more accurate for simulated biopsy of repeat targets than was 2D or 3D transrectal US, with a biopsy accuracy of 0.86 mm +/- 0.47 (standard deviation), 3.68 mm +/- 2.60, and 3.60 mm +/- 2.57, respectively. Experts had a 70% probability of sampling a prior biopsy target volume of 0.5 cm(3) with 2D transrectal US; however, the probability approached 100% with guided 3D transrectal US. Biopsy accuracy was not significantly different between experts and residents for any modality; however, experts were significantly (P < .05) faster than residents with each modality. Repeat biopsy of the prostate with 2D transrectal US has limited accuracy. Compared with 2D transrectal US, the biopsy accuracy of both experts and residents improved with guided 3D transrectal US but did not improve with conventional 3D transrectal US.


Survey of wild mammal hosts of cutaneous leishmaniasis parasites in panamá and costa rica.
The eco-epidemiology of American cutaneous leishmaniasis (ACL) is driven by animal reservoir species that are a source of infection for sand flies that serve as vectors infecting humans with Leishmania spp parasites. The emergence and re-emergence of this disease across Latin America calls for further studies to identify reservoir species associated with enzootic transmission. Here, we present results from a survey of 52 individuals from 13 wild mammal species at endemic sites in Costa Rica and Panama where ACL mammal hosts have not been previously studied. For Leishmania spp. diagnostics we employed a novel PCR technique using blood samples collected on filter paper. We only found Leishmania spp parasites in one host, the two-toed sloth, Choloepus hoffmanni. Our findings add further support to the role of two-toed sloths as an important ACL reservoir in Central America.


Eight Mutations of Three Genes (EDA, EDAR, and WNT10A) Identified in Seven Hypohidrotic Ectodermal Dysplasia Patients.
Hypohidrotic ectodermal dysplasia (HED) is characterized by abnormal development of the teeth, hair, and sweat glands. Ectodysplasin A (EDA), Ectodysplasin A receptor (EDAR), and EDAR-associated death domain (EDARADD) are candidate genes for HED, but the relationship between WNT10A and HED has not yet been validated. In this study, we included patients who presented at least two of the three ectodermal dysplasia features. The four genes were analyzed in seven HED patients by PCR and Sanger sequencing. Five EDA and one EDAR heterozygous mutations were identified in families 1-6. Two WNT10A heterozygous mutations were identified in family 7 as a compound heterozygote. c.662G>A (p.Gly221Asp) in EDA and c.354T>G (p.Tyr118*) in WNT10A are novel mutations. Bioinformatics analyses results confirmed the pathogenicity of the two novel mutations. In family 7, we also identified two single-nucleotide polymorphisms (SNPs) that were predicted to affect the splicing of EDAR. Analysis of the patient's total RNA revealed normal splicing of EDAR. This ascertained that the compound heterozygous WNT10A mutations are the genetic defects that led to the onset of HED. Our data revealed the genetic basis of seven HED patients and expended the mutational spectrum. Interestingly, we confirmed WNT10A as a candidate gene of HED and we propose WNT10A to be tested in EDA-negative HED patients.


Tracking mobile users in wireless networks via semi-supervised colocalization.
Recent years have witnessed the growing popularity of sensor and sensor-network technologies, supporting important practical applications. One of the fundamental issues is how to accurately locate a user with few labeled data in a wireless sensor network, where a major difficulty arises from the need to label large quantities of user location data, which in turn requires knowledge about the locations of signal transmitters or access points. To solve this problem, we have developed a novel machine learning-based approach that combines collaborative filtering with graph-based semi-supervised learning to learn both mobile users' locations and the locations of access points. Our framework exploits both labeled and unlabeled data from mobile devices and access points. In our two-phase solution, we first build a manifold-based model from a batch of labeled and unlabeled data in an offline training phase and then use a weighted k-nearest-neighbor method to localize a mobile client in an online localization phase. We extend the two-phase colocalization to an online and incremental model that can deal with labeled and unlabeled data that come sequentially and adapt to environmental changes. Finally, we embed an action model to the framework such that additional kinds of sensor signals can be utilized to further boost the performance of mobile tracking. Compared to other state-of-the-art systems, our framework has been shown to be more accurate while requiring less calibration effort in our experiments performed on three different testbeds.


Paired-homeodomain transcription factor PAX4 acts as a transcriptional repressor in early pancreatic development.
The paired-homeodomain transcription factor PAX4 is expressed in the developing pancreas and along with PAX6 is required for normal development of the endocrine cells. In the absence of PAX4, the numbers of insulin-producing beta cells and somatostatin-producing delta cells are drastically reduced, while the numbers of glucagon-producing alpha cells are increased. To gain insight into PAX4 function, we cloned a full-length Pax4 cDNA from a beta-cell cDNA library and identified a bipartite consensus DNA binding sequence consisting of a homeodomain binding site separated from a paired domain binding site by 15 nucleotides. The paired half of this consensus sequence has similarities to the PAX6 paired domain consensus binding site, and the two proteins bind to common sequences in several islet genes, although with different relative affinities. When expressed in an alpha-cell line, PAX4 represses transcription through the glucagon or insulin promoters or through an isolated PAX4 binding site. This repression is not simply due to competition with the PAX6 transcriptional activator for the same binding site, since PAX4 fused to the unrelated yeast GAL4 DNA binding domain also represses transcription through the GAL4 binding site in the alpha-cell line and to a lesser degree in beta-cell lines and NIH 3T3 cells. Repressor activity maps to more than one domain within the molecule, although the homeodomain and carboxyl terminus give the strongest repression. PAX4 transcriptional regulation apparently plays a role only early in islet development, since Pax4 mRNA as determined by reverse transcriptase PCR peaks at embryonic day 13.5 in the fetal mouse pancreas and is undetectable in adult islets. In summary, PAX4 can function as a transcriptional repressor and is expressed early in pancreatic development, which may allow it to suppress alpha-cell differentiation and permit beta-cell differentiation.


DDX5 promotes hepatocellular carcinoma tumorigenesis via Akt signaling pathway.
The DEAD-box-protein DDX5 is an ATP-dependent RNA helicase and also acts as co-activator that contributes to progression and metastasis of various tumours. However, its expression as well as prognostic roles of DDX5 in hepatocellular carcinoma (HCC) remain elusive. In this study, we investigated clinical significance and biological functions of DDX5 in HCC. Our results suggested that DDX5 showed overexpression at both transcriptional and translational levels in HCC tissues compared with adjacent normal tissues. Moreover, DDX5 expression was demonstrated to be correlated with tumor size (p < 0.001), N stage (p = 0.013), M stage (p = 0.006), tumor differentiation (p < 0.001) and American Joint Committee on Cancer (AJCC) stage (p = 0.001). Simultaneously, high DDX5 expression was found to be significantly correlated to worse outcome including Disease free survival (DFS) (p = 0.016) and overall survival (OS) (p = 0.032) according to Kaplan-Meier survival analysis. In vitro studies, it suggested that knockdown of DDX5 suppressed HCC cells migration, invasion and epithelial -to- mesenchymal transition (EMT) process. Depletion of DDX5 could promote HCC cells growth. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that PI3K/Akt signaling pathway obtained the highest enrichment. Furthermore, we found that knockdown of DDX5 decreased Akt as well as p-Akt (S473) expressions. Collectively, these findings suggested that DDX5 facilitated HCC cells growth via Akt signaling pathway. DDX5 played a crucial role in HCC proliferation and tumorigenesis and may be a novel prognostic marker and potential therapeutic target for HCC.


Difference in Mono-O-Glucosylation of Ras Subtype GTPases Between Toxin A and Toxin B From Clostridioides difficile Strain 10463 and Lethal Toxin From Clostridium sordellii Strain 6018.
Clostridioides difficile toxin A (TcdA) and Toxin B (TcdB) trigger inflammasome activation with caspase-1 activation in cultured cells, which in turn induce the release of IL-6, IFN-γ, and IL-8. Release of these proinflammatory responses is positively regulated by Ras-GTPases, which leads to the hypothesis that Ras glucosylation by glucosylating toxins results in (at least) reduced proinflammatory responses. Against this background, data on toxin-catalyzed Ras glucosylation are required to estimate of pro-inflammatory effect of the glucosylating toxins. In this study, a quantitative evaluation of the GTPase substrate profiles glucosylated in human colonic (Caco-2) cells treated with either TcdA, TcdB, or the related Clostridium sordellii lethal toxin (TcsL) was performed using multiple reaction monitoring (MRM) mass spectrometry. (H/K/N)Ras are presented to be glucosylated by TcsL and TcdA but by neither TcdB isoform tested. Furthermore, the glucosylation of (H/K/N)Ras was detected in TcdA-(not TcdB)-treated cells, as analyzed exploiting immunoblot analysis using the Ras glucosylation-sensitive 27H5 antibody. Furthermore, [14C]glucosylation of substrate GTPase was found to be increased in a cell-free system complemented with Caco-2 lysates. Under these conditions, (H/K/N)Ras glucosylation by TcdA was detected. In contrast, TcdB-catalyzed (H/K/N)Ras glucosylation was detected by neither MRM analysis, immunoblot analysis nor [14C]glucosylation in a cell-free system. The observation that TcdA (not TcdB) glucosylates Ras subtype GTPases correlates with the fact that TcdB (not TcdA) is primarily responsible for inflammatory responses in CDI. Finally, TcsL more efficaciously glucosylated Ras subtype GTPase as compared with TcdA, reinforcing the paradigm that TcsL is the prototype of a Ras glucosylating toxin.


Intracellular galectins in cancer cells: potential new targets for therapy (Review).
Dysregulation of galectin expression is frequently observed in cancer tissues. Such an abnormal expression pattern often correlates with aggressiveness and relapse in many types of cancer. Because galectins have the ability to modulate functions that are important for cell survival, migration and metastasis, they also represent attractive targets for cancer therapy. This has been well-exploited for extracellular galectins, which bind glycoconjugates expressed on the surface of cancer cells. Although the existence of intracellular functions of galectins has been known for many years, an increasing number of studies indicate that these proteins can also alter tumor progression through their interaction with intracellular ligands. In fact, in some instances, the interactions of galectins with their intracellular ligands seem to occur independently of their carbohydrate recognition domain. Such findings call for a change in the basic assumptions, or paradigms, concerning the activity of galectins in cancer and may force us to revisit our strategies to develop galectin antagonists for the treatment of cancer.


A pilot study of a supervised group exercise programme as a rehabilitation treatment for women with breast cancer receiving adjuvant treatment.
This pilot study examined whether exercise as an adjunctive rehabilitation therapy could benefit women who have early stage breast cancer and are currently receiving chemotherapy/radiotherapy. The study was designed as a randomised controlled trial (RCT). Physical functioning, fatigue and Quality of Life (QoL) outcomes were evaluated pre and post a 12-week intervention. The results showed that after 12 weeks the women who participated in the exercise programme (n = 12) displayed significantly higher levels of physical functioning and reported higher QoL scores than the controls (n = 10). Changes in fatigue and satisfaction with life favoured the intervention group but did not reach significance. These results are encouraging and suggest that a structured group exercise programme during adjuvant treatment is a safe, well tolerated and effective way of providing physical and psychological health benefits to women during treatment for early stage breast cancer. Since this was a pilot study the numbers did not allow appropriately powered analyses of some variables of interest and favoured relatively young and socio-economically advantaged women. Future studies need to address these issues and determine if these short-term benefits can be sustained.


Identification of gangliosides recognized by IgG anti-GalNAc-GD1a antibodies in bovine spinal motor neurons and motor nerves.
The presence of immunoglobulin G (IgG)-type antibodies to the ganglioside, N-acetylgalactosaminyl GD1a (GalNAc-GD1a), is closely associated with the pure motor type of Guillain-Barré syndrome (GBS). In the present study, we isolated disialogangliosides from the motor neurons and motor nerves of bovine spinal cords by DEAE-Sephadex column chromatography. The disialoganglioside fraction contained GD1a, GD2, GD1b, and three gangliosides, designated X1, X2 and X3. Serum from a patient with axonal GBS with IgG anti-GalNAc-GD1a antibody yielded positive immunostaining with X1, X2, and X3. When isolated by preparative thin-layer chromatography (TLC), X1 migrated at the same position as GalNAc-GD1a from Tay-Sachs brain, suggesting that X1 is GalNAc-GD1a containing N-acetylneuraminic acid (NeuAc). TLC of isolated X2 revealed that it migrated between GD1a and GD2. On the other hand, X3 had a migratory rate on TLC between and GD1b and GT1b. Since both X2 and X3 were recognized by IgG anti-GalNAc-GD1a antibody, the results suggest that X2 is a GalNAc-GD1a species containing a mixture containing a NeuAc-and an N-glycolylneuraminic acid (NeuGc) species, and X3 is a GalNAc-GD1a species with two NeuGc. This evidence indicating the specific localization of GalNAc-GD1a and its isomers in spinal motor neurons should be useful in elucidating the pathogenic role of IgG anti-GalNAc-GD1a antibody in pure motor-type GBS.


Applicability of short hydrogen breath test for screening of lactose malabsorption.
The gold standard for diagnosing lactose malabsorption is the H2 hydrogen breath test (HBT). Different methods of HBT have been proposed. However, in clinical practice the HBT is often shortened to 1-2 hr without proper validation. Our objective was to establish whether the usefulness of the HBT is influenced by shortening of the test and/or by substrate variations. In 62 patients with clinically suspected lactose intolerance and a positive lactose HBT we calculated the sensitivity of the HBT depending on the duration of the HBT. To determine whether substrate variations influence the sensitivity of the HBT, in another group of 32 patients with clinically suspected lactose intolerance and a positive milk HBT, the sensitivity of the HBT was also calculated depending on the duration of the test after milk ingestion. In other unselected 97 individuals, the result of the HBT with 360 ml of whole milk supplemented with lactose was compared with a symptomatic score for lactose intolerance to evaluate the specificity of the shortened milk HBT. Breath H2 excretion was significantly higher after lactose than after milk load (P < 0.01), and the increase in H2 appeared earlier with lactose than with milk (60 vs 90, min respectively). HBT duration influenced the sensitivity of the test that decreased from 95% for the 3-hr HBT to 37% for the 1-hr HBT with lactose and from 80% for 3-hr HBT to 21% for 1-hr HBT with milk. The specificity was similar for the 3-hr milk HBT and the 5-hr test (67 vs 62%). In conclusion, for screening of lactose malabsorption, the HBT can be shortened to 3 hr without loss of sensitivity and specificity, when a high dose of lactose load is used.


Quorum sensing coordinates brute force and stealth modes of infection in the plant pathogen Pectobacterium atrosepticum.
Quorum sensing (QS) in vitro controls production of plant cell wall degrading enzymes (PCWDEs) and other virulence factors in the soft rotting enterobacterial plant pathogen Pectobacterium atrosepticum (Pba). Here, we demonstrate the genome-wide regulatory role of QS in vivo during the Pba-potato interaction, using a Pba-specific microarray. We show that 26% of the Pba genome exhibited differential transcription in a QS (expI-) mutant, compared to the wild-type, suggesting that QS may make a greater contribution to pathogenesis than previously thought. We identify novel components of the QS regulon, including the Type I and II secretion systems, which are involved in the secretion of PCWDEs; a novel Type VI secretion system (T6SS) and its predicted substrates Hcp and VgrG; more than 70 known or putative regulators, some of which have been demonstrated to control pathogenesis and, remarkably, the Type III secretion system and associated effector proteins, and coronafacoyl-amide conjugates, both of which play roles in the manipulation of plant defences. We show that the T6SS and a novel potential regulator, VirS, are required for full virulence in Pba, and propose a model placing QS at the apex of a regulatory hierarchy controlling the later stages of disease progression in Pba. Our findings indicate that QS is a master regulator of phytopathogenesis, controlling multiple other regulators that, in turn, co-ordinately regulate genes associated with manipulation of host defences in concert with the destructive arsenal of PCWDEs that manifest the soft rot disease phenotype.


Polypharmacy Management in Older Patients.
Medications to treat disease and extend life in our patients often amass in quantities, resulting in what has been termed "polypharmacy." This imprecise label usually describes the accumulation of 5, and often more, medications. Polypharmacy in advancing age frequently results in drug therapy problems related to interactions, drug toxicity, falls with injury, delirium, and nonadherence. Polypharmacy is associated with resulting increased hospitalizations and higher costs of care for individuals and health care systems. To reduce polypharmacy, we delineate a systematic, consultative approach to identify highest-risk medications and drug-therapy problems. We address strategic reductions (deprescribing) of medications in palliative care, long-term care, and ambulatory older adults. Best practices for reducing opioids, benzodiazepines, and other high-risk medications include education about risk and agreement by patients and their families, advocates, and care teams. Addressing deprescribing should be within the framework of patients' health status as their care and goals transition from longevity to a plan of maintaining alertness, comfort, and satisfaction of quality of life. A team approach to address polypharmacy and avoidance of high-risk therapy is optimal within long-term care. Patients with terminal illnesses or those moving toward a comfort-care emphasis benefit from medication adjustments that are recognized beneficially within each patient's care goals. In caring for older adults, the acknowledgement that complicated regimens and high-risk medications requires a care plan to reduce or prevent medication-related problems and costs that are associated with polypharmacy.


Use of a customized vision model to analyze the effects of higher-order ocular aberrations and neural filtering on contrast threshold performance.
Customized optical filtering and light-dependent neural filtering were implemented in an ideal-observer model for an L-alternative forced-choice visual task. The model was applied to a contrast threshold visual task with adaptive optics correction of ocular higher-order (HO) aberrations under different light regimes, for which experimental data have previously been obtained (J. Mod. Opt.55, 791, 2008). A separability measure was used to assess the model-observer performance and to investigate the joint effect of optical and neural filtering. The numerical results were consistent with the experimental data in the assessment of the effect of HO aberrations as a function of light level.


Deregulation of XPC and CypA by cyclosporin A: an immunosuppression-independent mechanism of skin carcinogenesis.
Skin cancer is the most common malignancy in organ transplant recipients, causing serious morbidity and mortality. Preventing and treating skin cancer in these individuals has been extraordinarily challenging. Following organ transplantation, cyclosporin A (CsA) has been used as an effective immunosuppressive to prevent rejection. Therefore immunosuppression has been widely assumed to be the major cause for increased skin carcinogenesis. However, the mechanism of skin carcinogenesis in organ transplant recipients has not been understood to date; specifically, it remains unknown whether these cancers are immunosuppression dependent or independent. Here, using both immunocompromised nude mice which are defective in mature T lymphocytes as an in vivo model and human keratinocytes as an in vitro model, we showed that CsA impairs genomic integrity in the response of keratinocytes to ultra violet B (UVB). Following UVB radiation, CsA inhibited UVB-induced DNA damage repair by suppressing the transcription of the DNA repair factor xeroderma pigmentosum C (XPC). In addition, CsA compromised the UVB-induced checkpoint function by upregulating the molecular chaperone protein cyclophilin A (CypA). XPC mRNA levels were lower, whereas CypA mRNA and protein levels were higher in human skin cancers than in normal skin. CsA-induced phosphoinositide 3-kinase(PI3K)/AKT activation was required for both XPC suppression and CypA upregulation. Blocking UVB damage or inhibiting the PI3K/AKT pathway prevented CsA-sensitized skin tumorigenesis. Our findings identified deregulation of XPC and CypA as key targets of CsA, and UVB damage and PI3K/AKT activation as two principal drivers for CsA-sensitized skin tumorigenesis, further supporting an immunosuppression-independent mechanism of CsA action on skin tumorigenesis.


Choristoneura fumiferana nucleopolyhedrovirus encodes a functional 3'-5' exonuclease.
The Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) encodes an ORF homologous to type III 3'-5' exonucleases. The CfMNPV v-trex ORF was cloned into the Bac-to-Bac baculovirus expression-vector system, expressed in insect Sf21 cells with an N-terminal His tag and purified to homogeneity by using Ni-NTA affinity chromatography. Biochemical characterization of the purified V-TREX confirmed that this viral protein is a functional 3'-5' exonuclease that cleaves oligonucleotides from the 3' end in a stepwise, distributive manner, suggesting a role in proofreading during viral DNA replication and DNA repair. Enhanced degradation of a 5'-digoxigenin- or 5'-(32)P-labelled oligo(dT)(30) substrate was observed at increasing incubation times or increased amounts of V-TREX. The 3'-excision activity of V-TREX was maximal at alkaline pH (9.5) in the presence of 5 mM MgCl(2), 2 mM dithiothreitol and 0.1 mg BSA ml(-1).


Vasoregulatory Autoantibodies and Clinical Outcome After Ischemic Stroke-PROSCIS-B.
Vasoregulatory autoantibodies including autoantibodies targeting G-protein-coupled receptors might play a functional role in vascular diseases. We investigated the impact of vasoregulatory autoantibodies on clinical outcome after ischemic stroke. Data were used from the PROSCIS-B (Prospective Cohort With Incident Stroke-Berlin). Autoantibody-targeting receptors such as angiotensin II type 1 receptor (AT1R), endothelin-1 type A receptor, complement factor-3 and -5 receptors, vascular endothelial growth factor receptor-1 and -2, vascular endothelial growth factor A and factor B were measured. We explored associations of high antibody levels with (1) poor functional outcome defined as modified Rankin Scale >2 or Barthel Index <60 at 1 year after stroke, (2) Barthel Index scores over time using general estimating equations, and (3) secondary vascular events (recurrent stroke, myocardial infarction) or death up to 3 years using Cox proportional hazard models. We included 491 patients with ischemic stroke with data on autoantibody levels and outcome. In models adjusted for demographics and vascular risk factors, high autoantibody concentrations (quartile 4) targeting complement factor C3a receptor, vascular endothelial growth factor receptor-2, and vascular endothelial growth factor B were associated with poor functional outcome at 1 year: (odds ratio, 2.0 [95% CI, 1.1-3.6]; odds ratio, 1.8 [95% CI, 1.1-3.2]; and odds ratio, 2.1 [95% CI, 1.2-3.6], respectively) and with lower Barthel Index scores over 3 years (complement factor C3a receptor: adjusted β=-3.3 [95% CI, -5.7 to -0.5]; VEGF-B: adjusted β=-2.4 [95% CI, -4.8 to -0.06]). Patients with high autoantibody levels were not at higher risk for secondary vascular events or death. High levels of autoantibodies against vascular endothelial growth factor receptor-2, vascular endothelial growth factor B, and complement factor C3a receptor measured are associated with poor functional outcome after stroke but not with recurrent vascular events or death. URL: https://www.clinicaltrials.gov; Unique identifier: NCT01363856.


Measurement of the time course of optical quality and visual deterioration during tear break-up.
To compare changes in optical quality and visual performance that accompany tear break-up (TBU) during blink suppression. A three-channel optical system was developed that simultaneously measured refractive aberrations (Shack-Hartmann aberrometer), 20/40 letter contrast sensitivity (CS), and TBU (retroillumination, RI). Ten wearers of silicone hydrogel contact lenses were asked to keep one eye open for approximately 18 seconds, while CS, wavefront aberrations, and RI images were collected. The wavefront was reconstructed by zonal methods, and image quality was quantified with a series of metrics including RMS fit error. Novel metrics for quantifying TBU over the contact lens surface were developed by quantifying the contrast of the RI image and by using Fourier descriptors of the first Purkinje (PJ) image shape. There was a full range of TBU over the lens surface, with four subjects showing TBU across the corneal center and one subject with TBU in the inferior peripheral pupil. Among the four subjects with central corneal TBU, RMS fit error, RI contrast, and PJ Fourier descriptors showed high correlation with CS (r(2) range, 0.9187-0.9414, 0.6261-0.975, and 0.4917-0.8986, respectively). Some of the general optical-quality metrics such as blur strength, neural sharpness, and area of modulation transfer function (MTF) also showed that change correlated with CS loss. Optical metrics of tear quality and retinal image quality are associated with the decline in vision that occurs with TBU. The evidence supports the hypothesis that blurry vision symptoms reported by contact lens wearers are caused by poor quality of the retinal image due to TBU.


Purification of murine adipocyte lipid-binding protein. Characterization as a fatty acid- and retinoic acid-binding protein.
An adipose-specific protein has been purified from murine 3T3-L1 adipocytes to greater than 98% homogeneity. A purification procedure was developed utilizing a combination of gel filtration, cation exchange chromatography, and covalent chromatography on activated-thiol Sepharose 4B. The protein exists as a single polypeptide with a molecular weight of about 15,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein contains 2 mol of reduced sulfhydryl groups per mol of protein and an amino terminus blocked to sequencing. Automated Edman degradation of trypsin and CNBr-derived peptides has verified that the purified protein is that predicted by the mRNA (Bernlohr, D. A., Angus, C. W., Lane, M. D., Bolanowski, M. A., and Kelly, T. J. Jr. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 5468-5472). Based on sequence analysis, the 15-kDa adipocyte protein is considered to be a member of a family of tissue-specific, cytosolic lipid-binding proteins. Utilizing a liposome assay, the purified protein binds both oleic acid and retinoic acid saturably with approximately 1 mol of ligand bound per mol of protein. Dissociation constants determined from Scatchard analysis were 3 and 50 microM, respectively. This report represents the first demonstration of a member of this family of structurally related proteins that is capable of binding both fatty acid and retinoic acid. Hence, we propose the name adipocyte lipid-binding protein, or ALBP.


CRISPR/Cas9-Induced fad2 and rod1 Mutations Stacked With fae1 Confer High Oleic Acid Seed Oil in Pennycress (Thlaspi arvense L.).
Pennycress (Thlaspi arvense L.) is being domesticated as an oilseed cash cover crop to be grown in the off-season throughout temperate regions of the world. With its diploid genome and ease of directed mutagenesis using molecular approaches, pennycress seed oil composition can be rapidly tailored for a plethora of food, feed, oleochemical and fuel uses. Here, we utilized Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology to produce knockout mutations in the FATTY ACID DESATURASE2 (FAD2) and REDUCED OLEATE DESATURATION1 (ROD1) genes to increase oleic acid content. High oleic acid (18:1) oil is valued for its oxidative stability that is superior to the polyunsaturated fatty acids (PUFAs) linoleic (18:2) and linolenic (18:3), and better cold flow properties than the very long chain fatty acid (VLCFA) erucic (22:1). When combined with a FATTY ACID ELONGATION1 (fae1) knockout mutation, fad2 fae1 and rod1 fae1 double mutants produced ∼90% and ∼60% oleic acid in seed oil, respectively, with PUFAs in fad2 fae1 as well as fad2 single mutants reduced to less than 5%. MALDI-MS spatial imaging analyses of phosphatidylcholine (PC) and triacylglycerol (TAG) molecular species in wild-type pennycress embryo sections from mature seeds revealed that erucic acid is highly enriched in cotyledons which serve as storage organs, suggestive of a role in providing energy for the germinating seedling. In contrast, PUFA-containing TAGs are enriched in the embryonic axis, which may be utilized for cellular membrane expansion during seed germination and seedling emergence. Under standard growth chamber conditions, rod1 fae1 plants grew like wild type whereas fad2 single and fad2 fae1 double mutant plants exhibited delayed growth and overall reduced heights and seed yields, suggesting that reducing PUFAs below a threshold in pennycress had negative physiological effects. Taken together, our results suggest that combinatorial knockout of ROD1 and FAE1 may be a viable route to commercially increase oleic acid content in pennycress seed oil whereas mutations in FAD2 will likely require at least partial function to avoid fitness trade-offs.


Minimally permutated peptide analogs as internal standards for relative and absolute quantification of peptides and proteins.
A novel type of isobaric internal peptide standard for quantitative proteomics is described. The standard is a synthetic peptide derived from the target peptide by positional permutation of two amino acids. This type of internal standard is denominated minimally permutated peptide analog (MIPA). MIPA can be differentiated from their target analytes by LC-MS due to individual retention times and/or by MS/MS due to specific fragment ions. Both quantification methods are demonstrated using peptide mixtures of low and high complexity.


The Effect of Topical Antibiotic Powder Application in the Emergency Department on Deep Fracture-Related Infection in Type III Open Lower Extremity Fractures.
Despite advances in management, open fractures are at an elevated risk for deep fracture-related infection (FRI). Time to systemic antibiotic (ABX) administration and intraoperative topical administration of ABX powder have been used to decrease FRI risk. The purpose of this study was to determine whether topical application of antibiotic powder to type III open lower extremity fractures immediately on presentation to the emergency department (ED) reduces the rate of FRI. Prospective cohort compared with retrospective historical control. Level I trauma center. Application of 1 g of vancomycin and 1.2 g of tobramycin powder directly to open fracture wounds on presentation to the ED. Patients with type III open lower extremity fractures treated from July 1, 2019, to September 17, 2022, who received topical ABX powder in the ED were compared with patients from a 4-year historical cohort from July 1, 2015, to June 30, 2019, who were treated without topical ABX powder. Development of a FRI within 6 months of follow-up. Patient demographics, injury characteristics, and postoperative data were analyzed as risk factors for FRI. Sixty-six patients received topical ABX powder in the ED and were compared with 129 patients who were treated without topical ABX powder. The rate of FRI in the trial group was 6/66 (9.09%) versus 22/129 (17.05%) in the control cohort ( P = 0.133). Multivariate analysis demonstrated higher body mass index as a risk factor for development of FRI ( P = 0.036). No statistically significant difference in rates of FRI in open lower extremity fractures treated with immediate topical ABX administration in the ED versus standard-of-care treatment without topical ABX was found. These findings may have been limited by insufficient power. Further large-scale study is warranted to determine the significance of topical antibiotic powder application in the ED. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


Effect of tongue reduction on articulation in children with Down syndrome.
Tongue-reduction surgery was completed on 18 children with Down syndrome. Changes in articulation were evaluated pre- and postoperatively and at a 6-month follow-up. No significant differences in the number of articulation errors were found. The postoperative and 6-month follow-up scores of the surgery group were compared to a nonsurgery (contrast) group (N = 9) who had received two evaluations 6 months apart. Again, no significant differences were found. A follow-up survey of parental ratings on articulation of the surgery and nonsurgery group revealed that there were no significant differences between the groups but that parents of all the children claimed that "speech" had improved over the 6-month time period. These findings suggest that tongue-reduction surgery has no effect on the articulation of sounds.


Vasoregulatory hormones and the hyperfiltration of diabetes.
The role of renal vasoregulatory hormones in the hyperfiltration of early insulin-dependent diabetes mellitus (IDDM) was studied by micropuncture methods in rats with streptozotocin-induced diabetes. Seven to ten days after streptozotocin injection, untreated diabetic rats had elevated glomerular filtration rate (GFR) and single-nephron glomerular filtration rate (SNGFR), compared with normal euvolemic rats. Infusion of indomethacin (5 mg/kg) markedly reduced urinary and proximal tubular fluid prostaglandin E2 (PGE2), but GFR and SNGFR did not change. In a second group, intrarenal infusion of aprotinin (1,000 kallikrein inhibitor units.min-1.kg-1) to inhibit kallikrein also had no effect on GFR or SNGFR. In a third group, intrarenal infusion of angiotensin II (ANG II, 0.1 microgram.min-1.kg-1) reduced GFR, renal plasma flow (RPF), SNGFR, and glomerular plasma flow rate (QA) to values close to those in normal euvolemic rats. Single-nephron filtration fraction rose significantly with ANG II, but glomerular pressure (PG) was unaltered. Tubular fluid PGE2 increased in response to ANG II. Saralasin infusion following ANG II returned GFR, RPF, SNGFR, and QA to supernormal levels, and PG fell. In chronically salt-loaded normal rats, the responses to intrarenal ANG II and saralasin were similar to those observed in the diabetic rats. We conclude that hyperfiltration in early IDDM is not dependent on intact renal PGE2 or bradykinin synthesis. The results with ANG II infusion indicate that pre- and postglomerular and glomerular contractile cells of the diabetic kidney are able to constrict in response to this hormone.


When high-volume PCI operators in high-volume hospitals move to lower volume hospitals-Do they still maintain high volume and quality of outcomes?
The aim of this quasi-experimental study was to examine whether high-volume percutaneous coronary intervention (PCI) operators still maintain high volume and quality of outcomes when they moved to lower volume hospitals. Systematic reviews have indicated that high-volume PCI operators and hospitals have higher quality outcomes. However, little is known on whether high PCI volume and high quality outcomes are mainly due to operator characteristics (i.e., skill and experience) and is portable across organizations or whether it is due to hospital characteristics (i.e., equipment, team, and management system) and is less portable. We used Taiwan National Health Insurance claims data 2000-2012 to identify 98 high-volume PCI operators, 10 of whom moved from one hospital to another during the study period. We compared the PCI volume, risk-adjusted mortality ratio, and major adverse cardiovascular event (MACE) ratio before and after moving. Of the 10 high-volume operators who moved, 6 moved from high- to moderate- or low-volume hospitals, with median annual PCI volumes (interquartile range) of 130 (117-165) in prior hospitals and 54 (46-84) in subsequent hospitals (the hospital the operator moved to), and the remaining 4 moved from high to high-volume hospitals, with median annual PCI volumes (interquartile range) of 151 (133-162) in prior hospitals and 193 (178-239) in subsequent hospitals. No significant differences were observed in the risk-adjusted mortality ratios and MACE ratios between high-volume operators and matched controls before and after moving. High-volume operators cannot maintain high volume when they moved from high to moderate or low-volume hospitals; however, the quality of care is maintained. High PCI volume and high-quality outcomes are less portable and more hospital bound.


Cross-reactive surface epitopes on chondroitin sulfate A-adherent Plasmodium falciparum-infected erythrocytes are associated with transcription of var2csa.
Malaria in pregnancy is associated with placental accumulation of Plasmodium falciparum-infected erythrocytes (IE) that adhere to chondroitin sulfate A (CSA). Adhesion is mediated by P. falciparum erythrocyte membrane protein 1 (PfEMP1), a variant parasite protein expressed on the surface of IE and encoded by var genes. Rabbit antiserum was generated against the CSA-adherent P. falciparum line CS2, in which the dominant var transcribed is var2csa, a relatively conserved var gene that has been associated with CSA adhesion. Anti-CS2 recognized genetically distinct CSA-adherent P. falciparum lines but not CD36-adherent parent lines. Reactivity with anti-CS2 correlated with the level of adhesion to CSA. Fluorescence-activated cell sorting according to binding of anti-CS2 showed reactivity was associated with CSA adhesion and transcription of var2csa. These data are consistent with the hypothesis that var2csa encodes a PfEMP1 expressed on the surface of IE, which mediates adhesion to CSA and is relatively conserved between genetically distinct strains of P. falciparum.


Identifying Frequently Endorsed Benefits and Barriers to Breast Cancer Screening for African-Born Women in the NYC Metropolitan Area: a Pilot Study.
Most cancer screening data report on Black participants without distinguishing nativity, limiting our understanding of the needs of distinct groups within the African diaspora. The purpose of this pilot study was to assess demographic characteristics and perceptions of the benefits of and barriers to mammography among African immigrant women in New York City (NYC). Forty-two women who were 40 years or older, born in Africa, and English and/or French-speaking were recruited from African immigrant communities in NYC to complete a survey. Eighty percent of our sample aged 50 to 73 was adherent to the 2016 USPSTF mammography screening guideline. The most frequently endorsed benefits were that mammography will help find breast cancer early, could help find a breast lump before it is big enough to feel, and that if found early, breast cancer could be successfully treated. The most endorsed barriers were that having a mammogram is painful and that lack of insurance or being treated rudely at the mammogram center would keep participants from having a mammogram. Chi-square analyses assessed relationships between demographic characteristics and perceptions about mammography and revealed that endorsement of barriers to screening (e.g., health issues, transportation problems, pain, and time associated with mammography) varied by educational attainment. Findings suggest that future interventions should be multi-level and (1) support patients in accessing screening via resource sharing, (2) address other commonly cited barriers such as fear of pain during the procedure, and (3) support anti-racist healthcare environments especially in terms of treatment by providers.


Toward a sustainable mitigation approach of energy efficiency to greenhouse gas emissions in the European countries.
The efficient use of energy contributes to less energy consumption and the reduction of greenhouse gases released to nature, thus improving environmental sustainability. For this reason, many countries pioneered by the developed nations are trying to develop policies for energy efficiency. In this context, the relationship between energy efficiency and greenhouse gas emissions was tested by panel co-integration, panel causality, and FMOLS and DOLS analysis. Given that the study used the datasets of 29 European countries over the period 1995-2016, there result suggests that there is a long-term relationship between energy efficiency and greenhouse gas emissions and that the quantity of greenhouse gas emission decreases as energy efficiency increases. Finally, the robustness and novelty by employing the Emirmahmutoglu & Kose (2011) Testing for Granger causality in heterogeneous mixed panels. Economic Modelling, 28(3), 870-876 approach, the findings illustrated that there is a causal relationship between energy efficiency and greenhouse gas emissions for many European countries. Overall, the current study presents a relevant policy direction for the European bloc countries.


[Current possibilities in the outpatient treatment of postmyocardial infarction].
The lecture is intended for primary health care physicians and dedicated to the comprehensive rehabilitation and follow-up of postmyocardial infarction patients at an outpatient stage. The lecture material is based on the results of analyzing the Russian and foreign literature and on the author's experience. The section on nondrug treatment for myocardial infarction attaches particular significance to the exposure of risk factors for the disease. Long-term drug prevention of recurrences at the present level is examined.


Stent fracture in the coronary and peripheral arteries.
Inherent risks of stenting include restenosis and thrombosis. Recently, stent fractures have been recognized as a complication that may result in thrombosis, perforation, restenosis, and migration of the stent resulting in morbidity and mortality. Stent fractures were originally seen in the superficial femoral arteries but have since then been reported in almost all vascular sites including the coronary, renal, carotid, iliac, and femoropopliteal arteries. Fractures are the result of the complex interplay between stent manufacturing, the stented segment, pulsatile and nonpulsatile biomechanical forces, and plaque morphology at a particular vascular site. The presentation of a patient with a fracture is highly variable, ranging from asymptomatic in nature, detected on routine screening without any sequelae, to sudden cardiac death related to a thrombosed coronary artery. Despite being recognized as an important complication, consensus on routine surveillance and diagnostic methods to detect fractures continues to be lacking. Fortunately, most cases are relatively benign and can be managed conservatively if detected. In the setting of recurrent symptoms, further intervention is usually sought. In review of the literature most cases are managed with placement of a stent over the fractured area, the stent-in-stent technique, but several other alternatives may be available. As the knowledge of the variables that make stents prone to fracture are identified, better technologies and techniques can be employed to minimize the risk of this complication. This article reviews the available literature on stent fractures and complications using data found on PubMed, MEDLINE, the Manufacturer and User Facility Device Experience (MAUDE) database, and the Cochrane databases.


Cytoskeletal architecture and components involved in the attachment of Trypanosoma congolense epimastigotes.
Scanning and transmission electron microscopy of Trypanosoma congolense epimastigotes attached to a plastic substratum shows them to elaborate a complex flagellum filament system and plaque with a highly organized structure. Non-ionic detergent extraction of these cells shows that the resulting cytoskeletons remain attached to the plaque. The subpellicular corset of microtubules can be removed by salt or Ca2+ treatment leaving the axoneme, paraflagellar rod, associated filaments and the plaque. Neither of these treatments therefore removed the plaque-associated material from the substratum. Analysis of these fractions by SDS-polyacrylamide gel electrophoresis reveals an abundant 70 kDa protein that is highly enriched in the salt extracted 'minimal plaque' structures and appears likely to be a major constituent of this structure. These studies reveal that the complex filament and microtubule systems of the cytoskeleton involved the attachment of trypanosomes to substrata and have established a method of biochemical fractionation of the structures and components involved.


Comparison of four methods for bioavailability-based risk assessment of mixtures of Cu, Zn, and Ni in freshwater.
Although chemical risk assessment is still mainly conducted on a substance-by-substance basis, organisms in the environment are typically exposed to mixtures of substances. Risk assessment procedures should therefore be adapted to fit these situations. Four mixture risk assessment methodologies were compared for risk estimations of mixtures of copper (Cu), zinc (Zn), and nickel (Ni). The results showed that use of the log-normal species sensitivity distribution (SSD) instead of the best-fit distribution and sampling species sensitivities independently for each metal instead of using interspecies correlations in metal sensitivity had little impact on risk estimates. Across 4 different monitoring datasets, between 0% and 52% of the target water samples were estimated to be at risk, but only between 0% and 15% of the target water samples were at risk because of the mixture of metals and not any single metal individually. When a natural baseline database was examined, it was estimated that 10% of the target water samples were at risk because of single metals or their mixtures; when the most conservative method was used (concentration addition [CA] applied directly to the SSD, i.e., CASSD ). However, the issue of metal mixture risk at geochemical baseline concentrations became relatively small (2% of target water samples) when a theoretically more correct method was used (CA applied to individual dose response curves, i.e., CADRC ). Finally, across the 4 monitoring datasets, the following order of conservatism for the 4 methods was shown (from most to least conservative, with ranges of median margin of safety [MoS] relative to CASSD ): CASSD > CADRC (MoS = 1.17-1.25) > IADRC (independent action (IA) applied to individual dose-response curves; MoS = 1.38-1.60) > IASSD (MoS = 1.48-1.72). Therefore, it is suggested that these 4 methods can be used in a general tiered scheme for the risk assessment of metal mixtures in a regulatory context. In this scheme, the CASSD method could serve as a first (conservative) tier to identify situations with likely no potential risk at all, regardless of the method used (the sum toxic unit expressed relative to the 5% hazardous concentration [SumTUHC5 ] < 1) and the IASSD method to identify situations of potential risk, also regardless of the method used (the multisubstance potentially affected fraction of species using the IASSD method [msPAFIA,SSD ] > 0.05). The CADRC and IADRC methods could be used for site-specific assessment for situations that fall in between (SumTUHC5 > 1 and msPAFIA,SSD < 0.05). Environ Toxicol Chem 2017;36:2123-2138. © 2017 SETAC.


Ultralight three-dimensional boron nitride foam with ultralow permittivity and superelasticity.
Dielectrics with ultralow permittivity within 2 times that of air, excellent mechanical performance, and high thermal stability are highly attractive to many applications. However, since the finding of silica aerogels in the 1930s, no alternative ultralight porous dielectric with density below 10 mg/cm(3) has been developed. Here we present three-dimensional hierarchical boron nitride foam with permittivity of 1.03 times that of air, density of 1.6 mg/cm(3), and thermal stability up to 1200 °C obtained by chemical vapor deposition on a nickel foam template. This BN foam exhibits complete recovery after cyclic compression exceeding 70% with permittivity within 1.12 times that of air. Gathering all these exceptional characters, the BN foam should create a breakthrough development of flexible ultralow-permittivity dielectrics and ultralight materials.


Epithelial cell coculture models for studying infectious diseases: benefits and limitations.
Countless in vitro cell culture models based on the use of epithelial cell types of single lineages have been characterized and have provided insight into the mechanisms of infection for various microbial pathogens. Diverse culture models based on disease-relevant mucosal epithelial cell types derived from gastrointestinal, genitourinary, and pulmonary organ systems have delineated many key host-pathogen interactions that underlie viral, parasitic, and bacterial disease pathogenesis. An alternative to single lineage epithelial cell monoculture, which offers more flexibility and can overcome some of the limitations of epithelial cell culture models based on only single cell types, is coculture of epithelial cells with other host cell types. Various coculture models have been described, which incorporate epithelial cell types in culture combination with a wide range of other cell types including neutrophils, eosinophils, monocytes, and lymphocytes. This paper will summarize current models of epithelial cell coculture and will discuss the benefits and limitations of epithelial cell coculture for studying host-pathogen dynamics in infectious diseases.


Mitochondria-targeted metformins: anti-tumour and redox signalling mechanisms.
Reports suggest that metformin exerts anti-cancer effects in diabetic individuals with pancreatic cancer. Thus, metformin is currently being repurposed as a potential drug in cancer treatment. Studies indicate that potent metformin analogues are required in cancer treatment because of the low bioavailability of metformin in humans at conventional antidiabetic doses. We proposed that improved mitochondrial targeting of metformin by attaching a positively charged lipophilic triphenylphosphonium group will result in a new class of mitochondria-targeted metformin analogues with significantly enhanced anti-tumour potential. Using this approach, we synthesized various mitochondria-targeted metformin analogues with different alkyl chain lengths. Results indicate that the antiproliferative effects increased with increasing alkyl chain lengths (100-fold to 1000-fold). The lead compound, mito-metformin10, potently inhibited mitochondrial respiration through inhibition of complex I, stimulation of superoxide and hydrogen peroxide formation and activation of AMPK. When used in combination with ionizing radiation, mito-metformin10 acted as a radiosensitizer of pancreatic cancer cells. Because of the 1000-fold-higher potency of mitochondria-targeted metformin10, therapeutically effective plasma concentrations likely can be achieved in cancer patients.


The histaminergic system as a target for the prevention of obesity and metabolic syndrome.
The control of food intake and body weight is very complex. Key factors driving eating behavior are hunger and satiety that are controlled by an interplay of several central and peripheral neuroendocrine systems, environmental factors, the behavioral state and circadian rhythm, which all concur to alter homeostatic aspects of appetite and energy expenditure. Brain histamine plays a fundamental role in eating behavior as it induces loss of appetite and has long been considered a satiety signal that is released during food intake (Sakata et al., 1997). Animal studies have shown that brain histamine is released during the appetitive phase to provide a high level of arousal preparatory to feeding, but also mediates satiety. Furthermore, histamine regulates peripheral mechanisms such as glucose uptake and insulin function. Preclinical research indicates that activation of H1 and H3 receptors is crucial for the regulation of the diurnal rhythm of food consumption; furthermore, these receptors have been specifically recognized as mediators of energy intake and expenditure. Despite encouraging preclinical data, though, no brain penetrating H1 receptor agonists have been identified that would have anti-obesity effects. The potential role of the H3 receptor as a target of anti-obesity therapeutics was explored in clinical trials that did not meet up to the expectations or were interrupted (clinicaltrials.gov). Nonetheless, interesting results are emerging from clinical trials that evaluated the attenuating effect of betahistine (an H1 agonist/H3 antagonist) on metabolic side effects associated with chronic antipsychotics treatment. Aim of this review is to summarize recent results that suggest the clinical relevance of the histaminergic system for the treatment of feeding disorders and provide an up-to-date summary of preclinical research. This article is part of the Special Issue entitled 'Histamine Receptors'.


Percutaneous excision of difficult osteoid osteomas using intraoperative AIRO CT navigation: a preliminary report.
Osteoid osteomas are well known for intraoperative technical difficulties for localization and adequate excision, especially when they are not clearly visualized on plain radiographs or when they occur in difficult and inaccessible situations. Localization in the radiology suite and shifting the patient to the operating room can be cumbersome, and can result in errors because of lack of real-time imaging. In these scenarios, intraoperative navigation will be useful. We report a technique of intraoperative computed tomography (CT) navigation for localization and excision of osteoid osteomas of the long bones. Six patients (four femoral and two tibial lesions) with a radiological diagnosis of osteoid osteoma, in whom the nidus could not be visualized clearly on plain radiographs, were treated with this technique. Intraoperative CT navigation with AIRO was performed and the images were registered to the computer. The lesion was then localized and excised using a high-speed burr. All patients underwent postexcision on-table CT scans, which showed complete excision of the nidus. All patients became symptom free and are doing well at a minimum follow-up of 6 months. Intraoperative CT navigation helps to exactly localize the nidus and also helps to confirm complete excision of the nidus. This is a safe, effective and minimally invasive method to treat osteoid osteomas, particularly those that are not amenable to excision under C arm guidance.


[Innovation in mental health from the perspective of users: a community movement in the Northeast of Brazil].
This study, originating from a multidimensional analysis on the innovation category in a Community Mental Health Movement of Bom Jardim (Fortaleza, Brazil) that examines subjective and psychosocial dimensions, seeks to examine to what extent the work developed by this movement is effectively an innovative model in mental healthcare from the perspective of users of this service. The methodology adopted was a qualitative case study method based on a critical-interpretive approach employing techniques such as in-depth interviews associated with focus groups. The categorization of the discursive material was conducted using a model based on the four main dimensions of psychiatric reform, namely conceptual, technical, legal and socio-cultural aspects. The results reveal the following: an emphasis on the individual rather than focusing on the sickness; care as the basis for purposeful actions; the co-production of genuine dialogue with multiple stakeholders, and the construction of autonomy emphasizing individual potential. In conclusion, it can be said that healthcare practices developed by the Movement involve deconstructing the traditional model of mental healthcare, permitting new forms of citizenship, thereby contributing to deinstitutionalization.


Biologically indeterminate yet ordered promiscuous gene expression in single medullary thymic epithelial cells.
To induce central T-cell tolerance, medullary thymic epithelial cells (mTEC) collectively express most protein-coding genes, thereby presenting an extensive library of tissue-restricted antigens (TRAs). To resolve mTEC diversity and whether promiscuous gene expression (PGE) is stochastic or coordinated, we sequenced transcriptomes of 6,894 single mTEC, enriching for 1,795 rare cells expressing either of two TRAs, TSPAN8 or GP2. Transcriptional heterogeneity allowed partitioning of mTEC into 15 reproducible subpopulations representing distinct maturational trajectories, stages and subtypes, including novel mTEC subsets, such as chemokine-expressing and ciliated TEC, which warrant further characterisation. Unexpectedly, 50 modules of genes were robustly defined each showing patterns of co-expression within individual cells, which were mainly not explicable by chromosomal location, biological pathway or tissue specificity. Further, TSPAN8+ and GP2+ mTEC were randomly dispersed within thymic medullary islands. Consequently, these data support observations that PGE exhibits ordered co-expression, although mechanisms underlying this instruction remain biologically indeterminate. Ordered co-expression and random spatial distribution of a diverse range of TRAs likely enhance their presentation and encounter with passing thymocytes, while maintaining mTEC identity.


Rapid generation of functional nanovesicles from human trophectodermal cells for embryo attachment and outgrowth.
Extracellular vesicles (EVs) are important mediators of embryo attachment and outgrowth critical for successful implantation. While EVs have garnered immense interest in their therapeutic potential in assisted reproductive technology by improving implantation success, their large-scale generation remains a major challenge. Here, we report a rapid and scalable production of nanovesicles (NVs) directly from human trophectoderm cells (hTSCs) via serial mechanical extrusion of cells; these NVs can be generated in approximately 6 h with a 20-fold higher yield than EVs isolated from culture medium of the same number of cells. NVs display similar biophysical traits (morphologically intact, spherical, 90-130 nm) to EVs, and are laden with hallmark players of implantation that include cell-matrix adhesion and extracellular matrix organisation proteins (ITGA2/V, ITGB1, MFGE8) and antioxidative regulators (PRDX1, SOD2). Functionally, NVs are readily taken up by low-receptive endometrial HEC1A cells and reprogram their proteome towards a receptive phenotype that support hTSC spheroid attachment. Moreover, a single dose treatment with NVs significantly enhanced adhesion and spreading of mouse embryo trophoblast on fibronectin matrix. Thus, we demonstrate the functional potential of NVs in enhancing embryo implantation and highlight their rapid and scalable generation, amenable to clinical utility.


Prostate cancer screening in the fit Chilean elderly: a head to head comparison of total serum PSA versus age adjusted PSA versus primary circulating prostate cells to detect prostate cancer at initial biopsy.
Prostate cancer is predominately a disease of older men, with a median age of diagnosis of 68 years and 71% of cancer deaths occurring in those over 75 years of age. While prostate cancer screening is not recommended for men>70 years, fit elderly men with controlled comorbidities may have a relatively long life expectancy. We compare the use of age related PSA with the detection of primary malignant circulating prostate cells mCPCs to detect clinically significant PC in this population. All men undergoing PC screening with a PSA>4.0 ng/ml underwent TRUS 12 core prostate biopsy (PB). Age, PSA, PB results defined as cancer/no-cancer, Gleason, number of positive cores and percentage infiltration were registered. Men had an 8 ml blood sample taken for mCPC detection; mononuclear cells were obtained using differential gel centrifugation and mCPCs were identified using immunocytochemistry with anti-PSA and anti-P504S. A mCPC was defined as a cell expressing PSA and P504S; a positive test as at least one mCPC detected/sample. Diagnostic yields for subgroups were calculated and the number of avoided PBs registered. Esptein criteria were used to define small grade tumours. A total of 610 men underwent PB, 398 of whom were aged <70 yrs. Men over 70 yrs had: a higher median PSA, 6.24 ng/ml versus 5.59 ng/ml (p=0.04); and a higher frequency of cancer detected 90/212 (43%) versus 134/398 (34%) (p=0.032). Some 34/134 cancers in men<70 yrs versus 22/90 (24%) of men>70 yrs complied with criteria for active surveillance. CPC detection: 154/398 (39%) men<70 yrs were CPC (+), specificity for cancer 86%, sensitivity 88%, 14/16 with a false (-) result had a small low grade PC. In men>70 years, 88/212 (42%) were CPC (+); specificity 92%, sensitivity 87%, 10/12 with a false (-) had small low grade tumours. False (+) results were more common in younger men 36/154 versus 10/88 (p<0.02). With a PSA cutoff of 6.5 ng/ml, in men<70 yrs, 108 PB would be avoided, missing 56 cancers of which 48 were clinically significant. Using CPC detection, 124 biopsies would be avoided, missing only 2 clinically significant cancers. In men>70 yrs using a PSA>6.5 ng/ml would have resulted in 108 PB with 34 PC detected, of which 14(41%) were small low grade tumours. The use of CPC detection in the fit elderly significantly decreases the number of PBs without missing clinically significant cancers, indicating superiority to the use of age-related PSA.


The arterial vascularization of the stomach fundus of human fetuses.
We have investigated the existence, frequency and the course of arteries of the human stomach fundus by injection method. Our material obtained 52 human fetuses of the length range from 127 to 285 mm and the fetal age range from l5 to 28 weeks of gestation. It has been found, that the source of the arterial blood supply of this region are two arteries: the left gastric artery and splenic artery with its bigger branches: posterior gastric artery, superior polar artery and short gastric arteries. The source of arterial blood supply of the stomach fundus there have been two in 6 cases (11,5%) or three in 45 cases (86,6%) or four in 1 case (l,9%) of these arteries. We have analysed in detail the contribution and the range of the following arteries in the stomach quadrants. It has been shown, that the anterior medial quadrant has the best arterial blood supply and the anterior lateral has the worst one.


The purification and characterization of phosphonopyruvate hydrolase, a novel carbon-phosphorus bond cleavage enzyme from Variovorax sp Pal2.
Phosphonopyruvate hydrolase, a novel bacterial carbon-phosphorus bond cleavage enzyme, was purified to homogeneity by a series of chromatographic steps from cell extracts of a newly isolated environmental strain of Variovorax sp. Pal2. The enzyme was inducible in the presence of phosphonoalanine or phosphonopyruvate; unusually, its expression was independent of the phosphate status of the cell. The native enzyme had a molecular mass of 63 kDa with a subunit mass of 31.2 kDa. Activity of purified phosphonopyruvate hydrolase was Co2+-dependent and showed a pH optimum of 6.7-7.0. The enzyme had a Km of 0.53 mm for its sole substrate, phosphonopyruvate, and was inhibited by the analogues phosphonoformic acid, 3-phosphonopropionic acid, and hydroxymethylphosphonic acid. The nucleotide sequence of the phosphonopyruvate hydrolase structural gene indicated that it is a member of the phosphoenolpyruvate phosphomutase/isocitrate lyase superfamily with 41% identity at the amino acid level to the carbon-to-phosphorus bond-forming enzyme phosphoenolpyruvate phosphomutase from Tetrahymena pyriformis. Thus its apparently ancient evolutionary origins differ from those of each of the two carbon-phosphorus hydrolases that have been reported previously; phosphonoacetaldehyde hydrolase is a member of the haloacetate dehalogenase family, whereas phosphonoacetate hydrolase belongs to the alkaline phosphatase superfamily of zinc-dependent hydrolases. Phosphonopyruvate hydrolase is likely to be of considerable significance in global phosphorus cycling, because phosphonopyruvate is known to be a key intermediate in the formation of all naturally occurring compounds that contain the carbon-phosphorus bond.


Noise-induced hearing loss can alter neural coding and increase excitability in the central nervous system.
Responses of auditory neurons in the inferior colliculi of mice were studied longitudinally before and shortly after each animal was exposed to intense noise. Noise exposure caused expected losses in auditory sensitivity, but in 31 percent of the neurons studied, unexpected alterations of temporal patterns of action potentials were observed: certain suprathreshold stimuli that had evoked only transient "onset" responses or inhibition of spontaneous discharges prior to noise exposure came to elicit sustained excitation after exposure. Thus, noise-induced hearing loss can be associated with increases in neural responsivity and alterations of normal neural coding processes.


Native Triple Valve Endocarditis as Complication of Post-Abortal Sepsis.
Infective endocarditis (IE) is a highly morbid condition in pregnancy which poses both maternal and fetal risk. In majority of cases, endocarditis occurs only on single valve and usually occurs on valve with structural disease or prosthetic valve. Multi-valvular involvement is not common and so we report a case of native triple valve endocarditis as a complication of post abortal sepsis which was successfully treated medically.


Chronic diarrhoea: investigation, treatment and nursing care.
Chronic diarrhoea is a distressing symptom of a number of conditions. This article explains the assessment of patients at the initial outpatient visit through the various investigations and finally medical and surgical treatment. Emphasis is placed on the nursing management of chronic diarrhoea, particularly the treatment of physical effects such as dehydration and perianal skin soreness, and the psychological aspects of care.


Modulation of genotoxicity of oxidative mutagens by glycyrrhizic acid from Glycyrrhiza glabra L.
The chemopreventive effects of certain phytoconstituents can be exploited for their use as functional foods, dietary supplements and even as drugs. The natural compounds, acting as anti-genotoxic and free radical scavenging compounds, may serve as potent chemopreventive agents. These can inhibit DNA modulatory activities of mutagens and help preventing pathological processes. Present study on Glycyrrhiza glabra L., a promising medicinal plant, widely used in traditional medicine, focused on the bioassay-guided fractionation of its extracts for the isolation of certain phytochemicals with anti-genotoxic potential against oxidative mutagens. The methanol extract of Glycyrrhiza glabra rhizomes was subjected to column chromatography, and isolated fraction was evaluated for its anti-genotoxic and antioxidant potential using SOS chromotest, Comet assay, and DPPH radical scavenging assay. GLG fraction, which was characterized as Glycyrrhizic acid, inhibited the genotoxicity of oxidative mutagens viz., H(2)O(2) and 4NQOquite efficiently. In SOS chromotest, using E.coli PQ37 tester strain, it inhibited induction factor induced by H(2)O(2) and 4NQO by 75.54% and 71.69% at the concentration of 121.46 μM,respectively. In Comet assay, it reduced the tail moment induced by H(2)O(2) and 4NQO by 70.21% and 69.04%, respectively, at the same concentration in human blood lymphocytes. The isolated fraction also exhibited DPPH free radical scavenging activity and was able to scavenge 85.95% radicals at a concentration of 120 μM. Glycyrrhizic acid is a potential modulator of genotoxins as well as efficient scavenger of free radicals.


Applying Machine Learning and Point-Set Registration to Automatically Measure the Severity of Spinal Curvature on Radiographs.
Measuring the severity of the lateral spinal curvature, or Cobb angle, is critical for monitoring and making treatment decisions for children with adolescent idiopathic scoliosis (AIS). However, manual measurement is time-consuming and subject to human error. Therefore, clinicians seek an automated measurement method to streamline workflow and improve accuracy. This paper reports on a novel machine learning algorithm of cascaded convolutional neural networks (CNN) to measure the Cobb angle on spinal radiographs automatically. The developed method consisted of spinal column segmentation using a CNN, vertebra localization and segmentation using iterative vertebra body location coupled with another CNN, point-set registration to correct vertebra segmentations, and Cobb angle measurement using the final segmentations. Measurement performance was evaluated with the circular mean absolute error (CMAE) and percentage within clinical acceptance ([Formula: see text]) between automatic and manual measurements. Analysis was separated by curve severity to identify any potential systematic biases using independent samples Student's t-tests. The method detected 346 of the 352 manually measured Cobb angles (98%), with a CMAE of 2.8° and 91% of measurements within the 5° clinical acceptance. No statistically significant differences were found between the CMAEs of mild ([Formula: see text]), moderate (25°-45°), and severe ([Formula: see text]) groups. The average measurement time per radiograph was 17.7±10.2s, improving upon the estimated average of 30s it takes an experienced rater to measure. Additionally, the algorithm outputs segmentations with the measurement, allowing clinicians to interpret measurement results. The developed method measured Cobb angles on radiographs automatically with high accuracy, quick measurement time, and interpretability, suggesting clinical feasibility.


Multivalent bi-specific nanobioconjugate engager for targeted cancer immunotherapy.
Tumour-targeted immunotherapy offers the unique advantage of specific tumouricidal effects with reduced immune-associated toxicity. However, existing platforms suffer from low potency, inability to generate long-term immune memory and decreased activities against tumour-cell subpopulations with low targeting receptor levels. Here we adopted a modular design approach that uses colloidal nanoparticles as substrates to create a multivalent bi-specific nanobioconjugate engager (mBiNE) to promote selective, immune-mediated eradication of cancer cells. By simultaneously targeting the human epidermal growth factor receptor 2 (HER2) expressed by cancer cells and pro-phagocytosis signalling mediated by calreticulin, the mBiNE stimulated HER2-targeted phagocytosis and produced durable antitumour immune responses against HER2-expressing tumours. Interestingly, although the initial immune activation mediated by the mBiNE was receptor dependent, the subsequent antitumour immunity also generated protective effects against tumour-cell populations that lacked the HER2 receptor. Thus, the mBiNE represents a new targeted, nanomaterial-immunotherapy platform to stimulate innate and adaptive immunity and promote a universal antitumour response.


Dry eye disease: pathophysiology, classification, and diagnosis.
Dry eye disease (DED) is a multifactorial disorder of the tear film and ocular surface that results in eye discomfort, visual disturbance, and often ocular surface damage. Although recent research has made progress in elucidating DED pathophysiology, currently there are no uniform diagnostic criteria. This article discusses the normal anatomy and physiology of the lacrimal functional unit and the tear film; the pathophysiology of DED; DED etiology, classification, and risk factors; and DED diagnosis, including symptom assessment and the roles of selected diagnostic tests.


Accelerating the development of novel technologies and tools for the subcutaneous delivery of biotherapeutics.
Subcutaneous (SC) delivery of biotherapeutics is well established as a route of administration across many therapeutic areas and has been shown to be effective and well-tolerated. It can offer several advantages over intravenous administration. This notwithstanding, there remain critical development issues and knowledge gaps in SC drug delivery. To articulate and address these issues, the SC Drug Delivery and Development Consortium was convened in 2018 as a pre-competitive collaboration of industry experts in drug delivery, device development, and commercialization. In this review, we outline the Consortium's vision and mission in advancing the development of patient-centered biotherapeutics and establishing a collaborative organization that facilitates open sharing of information and gives voice to diverse viewpoints from SC experts across industries and disciplines. Additionally, we describe the current landscape and challenges associated with SC administration of therapeutic proteins (specifically monoclonal antibodies) and offer insights into potential solutions to these challenges within the context of 8 problem statements developed by the Consortium to highlight key gaps, unmet needs, and actionable issues. Current and future opportunities to accelerate progress in the field through technological advances and the development of drug delivery tools are also discussed.


A Perfusion-based Human Cadaveric Model for Management of Carotid Artery Injury during Endoscopic Endonasal Skull Base Surgery.
Objective To create and develop a reproducible and realistic training environment to prepare residents and trainees for arterial catastrophes during endoscopic endonasal surgery. Design An artificial blood substitute was perfused at systolic blood pressures in eight fresh human cadavers to mimic intraoperative scenarios. Setting The USC Keck School of Medicine Fresh Tissue Dissection Laboratory was used as the training site. Participants Trainees were USC neurosurgery residents and junior faculty. Main Outcome A 5-point questionnaire was used to assess pre- and posttraining confidence scores. Results High-pressure extravasation at normal arterial blood pressure mimicked real intraoperative internal carotid artery (ICA) injury. Residents developed psychomotor skills required to achieve hemostasis using suction, cottonoids, and muscle grafts. Questionnaire responses from all trainees reported a realistic experience enhanced by the addition of the perfusion model. Conclusions The addition of an arterial perfusion system to fresh tissue cadavers is among the most realistic training models available. This enables the simulation of rare intraoperative scenarios such as ICA injury. Strategies for rapid hemostasis and implementation of techniques including endoscope manipulation, suction, and packing can all be rehearsed via this novel paradigm.