[Co-culture with umbilical cord mesenchymal stem cells promotes the synthesis and mechnism of milk protein in bovine mammary epithelial cells].
To explore the possible mechanism of umbilical cord mesenchymal stem cels( UCMSCs) regulating milk protein synthesis in bovine mammary gland epithelial cells( BMECs). UCMSCs and BMECs were co-cultured by double-chamber TranswellTM,and other BMECs were cultured alone as a control group. Insulin like growth factor 1 receptor( IGF-1R) inhibitor AG1024 was used to treat cells. IGF-1,β casein( CSN2) and κ casein( CSN3) content in the supernatants were determined by ELISA; the relative expression abundance of Janus kinase and signal transducer and activator of transcription factor( JAK / STAT) signaling pathway-related genes were detected by quantitative real-time PCR( qRT-PCR); and after the cells were treated with JAK2 signal blocker AG490,qRT-PCR was performed to test the relative expression abundance of CSN2 and CSN3 mRNAs. CSN2, CSN3 synthetic quantity, and the relative expression abundance of CSN2,CSN3,JAK2,STAT5,E74-like ETS transcription factor 5( ELF5) mRNAs of the co-culture group were significantly higher than those of the control group. After treated with AG1024,the co-culture group showed remarkably decreased CSN2,CSN3 synthesis in BMECs as wel as the decreased relative expression abundance of CSN2,CSN3,JAK2,STAT5,ELF5 mRNAs. After blocked with AG490,the relative expression abundance of CSN2 and CSN3 mRNAs were reduced significantly in the co-culture group. In addition,the expressions of CSN2 and CSN3 mRNAs were inhibited significantly when both AG1024 and AG490 were supplemented. UCMSCs can mediate JAK2 / STAT5 signaling pathwayvia IGF-1,and increase the expressions of milk protein synthesis key genes in BMECs,thus promoteing the synthesis of milk protein.


The isolation of Campylobacter hyointestinalis from a pig in South Africa.
The isolation of a strain of Campylobacter hyointestinalis from a piglet is described. The animal originated from a farm where another animal showed signs of intestinal adenomatosis. The animal from which the isolation was made had diarrhoea, and an enteropathogenic Escherichia coli was also isolated. No pathological changes indicative of intestinal adenomatosis were detected in this animal.


cDNA cloning and expression of the Xenopus homologue of the neural adhesion molecule, contactin (F3/F11).
Contactin (F3/F11) is an immunoglobulin superfamily cell surface glycoprotein predominantly expressed in the nervous system. To examine the structure and tissue distribution of Xenopus contactin, a cDNA clone was isolated based on the amino acid sequences conserved among chicken and mammalian contactin proteins. The conceptual translate of the cDNA consists of 1005 amino acid residues that have 70% identity to those of chicken and mammalian contactin. Northern blot hybridization using a labeled cDNA fragment revealed specific expression of 6.5 kb mRNA in the brain. Monoclonal and polyclonal antibodies prepared to the recombinant Xenopus contactin peptides detected a single 135 kD band on Western blots of the brain and spinal cord extracts. Differential extraction and phosphatidylinositol-specific phospholipase C (PI-PLC) digestion experiments showed that the immunoreactive 135 kD proteins bind, at least in part to the membrane by GPI anchor. On brain tissue sections, strong contactin immunoreactivities were detected on nerve fibers of a subset of cerebral and cerebellar neurons. These results suggest that the basic structure and tissue distribution of Xenopus contactin are similar to those in other vertebrates.


Fluorescence resonance energy transfer analysis of merlin conformational changes.
Neurofibromatosis type 2 is an inherited autosomal disorder caused by biallelic inactivation of the NF2 tumor suppressor gene. The NF2 gene encodes a 70-kDa protein, merlin, which is a member of the ezrin-radixin-moesin (ERM) family. ERM proteins are believed to be regulated by a transition between a closed conformation, formed by binding of their N-terminal FERM domain and C-terminal tail domain (CTD), and an open conformation, in which the two domains do not interact. Previous work suggests that the tumor suppressor function of merlin is similarly regulated and that only the closed form is active. Therefore, understanding the mechanisms that control its conformation is crucial. We have developed a series of probes that measures merlin conformation by fluorescence resonance energy transfer, both as purified protein and in live cells. Using these tools, we find that merlin exists predominately as a monomer in a stable, closed conformation that is mediated by the central alpha-helical domain. The contribution from the FERM-CTD interaction to the closed conformation appears to be less important. Upon phosphorylation or interaction with an effector protein, merlin undergoes a subtle conformational change, suggesting a novel mechanism that modulates the interaction between the FERM domain and the CTD.


A four-component model of the action potential in mouse detrusor smooth muscle cell.
Detrusor smooth muscle cells (DSMCs) of the urinary bladder are electrically connected to one another via gap junctions and form a three dimensional syncytium. DSMCs exhibit spontaneous electrical activity, including passive depolarizations and action potentials. The shapes of spontaneous action potentials (sAPs) observed from a single DSM cell can vary widely. The biophysical origins of this variability, and the precise components which contribute to the complex shapes observed are not known. To address these questions, the basic components which constitute the sAPs were investigated. We hypothesized that linear combinations of scaled versions of these basic components can produce sAP shapes observed in the syncytium. The basic components were identified as spontaneous evoked junction potentials (sEJP), native AP (nAP), slow after hyperpolarization (sAHP) and very slow after hyperpolarization (vsAHP). The experimental recordings were grouped into two sets: a training data set and a testing data set. A training set was used to estimate the components, and a test set to evaluate the efficiency of the estimated components. We found that a linear combination of the identified components when appropriately amplified and time shifted replicated various AP shapes to a high degree of similarity, as quantified by the root mean square error (RMSE) measure. We conclude that the four basic components-sEJP, nAP, sAHP, and vsAHP-identified and isolated in this work are necessary and sufficient to replicate all varieties of the sAPs recorded experimentally in DSMCs. This model has the potential to generate testable hypotheses that can help identify the physiological processes underlying various features of the sAPs. Further, this model also provides a means to classify the sAPs into various shape classes.


[Screening for congenital toxoplasmosis: pregnancy outcome after prenatal diagnosis in 211 cases].
To emphasize the importance of follow-up after birth of infants with an antenatal diagnosis of congenital toxoplasmosis. Retrospective study from July 1987 through January 31 1995 on 211 women (214 fetuses) who had undergone ovular biopsy for toxoplasmosis seroconversion during pregnancy. Antenatal diagnosis was positive in 13 patients (6.2%). Four pregnancies were terminated during the second trimester. Delivery was triggered at 37 weeks gestation in one woman and in 8 others pregnancy was continued with Fansidar. All infants were born live with infraclinic disease. Two fetal deaths related to biopsy technique occurred (0.95%) and one pregnancy was terminated before the results had been obtained. Only 21.3% of the patients were delivered in our unit. A total of 197 infants were delivered with a negative antenatal diagnosis: 93 were healthy, 5 had congenital toxoplasmosis, 1 died at 3 months and 98 had no or incomplete follow-up. Incomplete post-natal follow-up is in contradiction with the excellent performance of antenatal diagnosis of congenital toxoplasmosis. Greater care is needed, especially since now only an amniocentisis is required to detect the parasite genome with polymerase chain reaction.


The type of leg lost affects habitat use but not survival in a non-regenerating arthropod.
Finding shelter and surviving encounters with predators are pervasive challenges for animals. These challenges may be exacerbated after individuals experience bodily damage. Certain forms of damage arise voluntarily in animals; for instance, some taxa release appendages (tails, legs, or other body parts) as a defensive strategy ("autotomy"). This behavior, however, may pose long-term negative consequences for habitat use and survival. Additionally, these putative consequences are expected to vary according to the function of the lost body part. We tested the effects of losing different functional leg types (locomotor or sensory) on future habitat use and survival in a Neotropical species of Prionostemma harvestmen (Arachnida: Opiliones) that undergo frequent autotomy but do not regrow limbs. Daytime surveys revealed that both eight-legged harvestmen and harvestmen missing legs roosted in similar frequencies across habitats (tree bark, mossy tree, or fern), and perched at similar heights. Mark-recapture data showed that harvestmen that lost sensory legs roosted in tree bark less frequently, but on mossy trees more frequently. On the contrary, we did not observe changes in habitat use for eight-legged animals or animals that lost locomotor legs. This change might be related to sensory exploration and navigation. Lastly, we found that recapture rates across substrates were not affected by the type of legs lost, suggesting that leg loss does not impact survival. This potential lack of effect might play a role in why a defensive strategy like autotomy is so prevalent in harvestmen despite the lack of regeneration.


The influence of time and irrigant refreshment on biofilm removal from lateral morphological features of simulated root canals.
To evaluate the effect of irrigant refreshment and exposure time of a 2% sodium hypochlorite solution (NaOCl) on biofilm removal from simulated lateral root canal spaces using two different flow rates. A dual-species biofilm was formed by a Constant Depth Film Fermenter (CDFF) for 96 h in plug inserts with anatomical features resembling an isthmus or lateral canal-like structures. The inserts were placed in a root canal model facing the main canal. NaOCl 2% and demineralized water (control group) were used as irrigant solutions. Both substances were applied at a flow rate of 0.05 and 0.1 mL s-1 . The samples were divided into three groups with zero, one or two refreshments in a total exposure time of 15 min. A three-way analysis of variance (anova) was performed to investigate the interaction amongst the independent variables and the effect of consecutive irrigant refreshment on percentage of biofilm removal. A Tukey post hoc test was used to evaluate the effect of each independent variable on percentage biofilm removal in the absence of statistically significant interactions. For the lateral canal, NaOCl removed significantly more biofilm irrespective of the number of refreshments and exposure time (P = 0.005). There was no significant effect in biofilm removal between the consecutive irrigant refreshments measured in the same biofilm. For the isthmus, NaOCl removed significantly more biofilm irrespective of the number of refreshments and exposure time; both NaOCl and a high flow rate removed significantly more biofilm when the exposure time was analysed (P = 0.018 and P = 0.029, respectively). Evaluating the effect of consecutive irrigant refreshment on the same biofilm, 2% NaOCl, 0.1 mL s-1 flow rate and one or two refreshments removed significant more biofilm (P = 0.04, 0.034 and 0.003, <0.001, respectively). In this model, refreshment did not improve biofilm removal from simulated lateral root canal spaces. NaOCl removed more biofilm from the lateral canal- and isthmus-like structure. A higher flow rate removed significantly more biofilm from the isthmus-like structure. There was always remaining biofilm left after the irrigation procedures.


Variation of the expression of Mycobacterium tuberculosis ppe44 gene among clinical isolates.
PPE44 is a member of the Mycobacterium tuberculosis PPE proteins, a polymorphic family of 69 glycine-rich proteins that predictively represent a source of antigenic variation. The genetic diversity of gene ppe44 among clinical isolates has been studied. No genomic polymorphism of ppe44 was found by a PCR-restriction fragment length polymorphism assay using three restriction enzymes. Nucleotide sequencing of gene ppe44 of a number of isolates, selected to represent the major phylogenetic lineages of M. tuberculosis, showed no nucleotide substitution, with the exception of isolates of the Beijing genotype. These findings indicate that gene ppe44 is basically conserved among M. tuberculosis strains. The expression of gene ppe44 was then determined at the transcriptional level by a real-time reverse transcriptase PCR assay. Extremely high quantitative variations in ppe44 expression were found among the isolates; ppe44 expression of the Beijing strains was significantly higher than the non-Beijing strains. To test whether differential expression of gene ppe44 has the potential to provide a dynamic antigen display, antibodies to PPE44 were titered in the sera of M. tuberculosis-infected subjects. Variation of antibody response to PPE44 was found with regard to both antibody titers and the proportion of responding subjects. These results indicate that the differential expression of genes ppe could influence the host's immune responsiveness, thus having implications in the immunopathogenesis of tuberculosis.


Regulation of Kappa Opioid Receptor Inactivation Depends on Sex and Cellular Site of Antagonist Action.
The prototypical member of the receptor-inactivating kappa opioid receptor (KOR) antagonists, norbinaltorphimine (norBNI), produces prolonged receptor inactivation by a cJun kinase mechanism. These antagonists have potential therapeutic utility in the treatment of stress disorders; however, additional preclinical characterization is necessary to understand important aspects of their action. In this study, we report that norBNI does not work as effectively in female mice as in males because of estrogen regulation of G protein receptor kinase (GRK); pretreatment of ovary-intact female mice with the selective GRK2/3 inhibitor, Compound 101, made females equally sensitive to norBNI as males. Prior observations suggested that in vivo treatment with norBNI does not produce long-lasting inhibition of KOR regulation of dopamine release in the nucleus accumbens. We assessed the persistence of norBNI receptor inactivation in subcellular compartments. Fast-scan cyclic voltammetry recordings confirmed that presynaptic inhibition of dopamine release by the KOR agonist U69,593 was not blocked by in vivo pretreatment with norBNI under conditions that prevented KOR-mediated aversion and analgesia. We employed a novel in vivo proxy sensor of KOR activation, adenovirus associated double floxed inverted-HyPerRed, and demonstrated that KOR activation stimulates cJun kinase-dependent reactive oxygen species (ROS) production in somatic regions of ventral tegmental area dopamine neurons, but did not activate ROS production in dopamine terminals. The compartment selective action helps explain how dopamine somatic, but not terminally expressed, KORs are inactivated by norBNI. These results further elucidate molecular signaling mechanisms mediating receptor-inactivating KOR antagonist action and advance medication development for this novel class of stress-resilience medications. SIGNIFICANCE STATEMENT: Kappa opioid receptor (KOR) antagonists are being developed as novel proresilience therapeutics for the treatment of mood and substance use disorders. This study showed that the long-acting KOR antagonists are affected by both the sex of the animal and the subcellular compartment in which the receptor is expressed.


Health status of Gypsy Travellers.
Although previous studies suggest that Gypsy Travellers have poorer health status and excess mortality compared with the general population, there is no epidemiological evidence using validated measures in this nomadic ethnic group. The aim of this study was to compare the health status of traditional Gypsy Travellers with norms from the UK population, and with a concurrent comparison group using the Euroqol health status measure (EQ-5D). Eighty-seven adult Gypsy Travellers were matched for age and sex with English or Irish residents, registered with an urban general practice in an area of high social deprivation. Both samples completed the EQ-5D questionnaire by interview. A comparison was also made with normative data from the UK general population. Travellers had poorer health status than their settled counterparts on two of the five dimensions (mobility and activity) but not on the overall summary score. Travellers reported significantly poorer health than the matched comparison group on the EQ-5D visual analogue scale. Both the Travellers and the comparison group had much poorer health status on the EQ-5D index than the UK population norms, even when compared with the lowest socioeconomic group. Health status of Gypsy Travellers was significantly poorer than in the lowest socio-economic UK population group, but was not so markedly different from a concurrent, matched, socially deprived resident group. Gypsy Travellers did have poorer health status than matched comparators in relation to mobility, activity and perception of overall health. Quantitative assessment of health status in the Traveller community is feasible.


Paraneoplastic Neurological Syndrome in Fallopian Tube Cancer.
We report on a rare case of paraneoplastic syndrome (PS) that was discovered on completion of diagnostic work-up to be an undifferentiated fallopian tube carcinoma. A 49-year-old Caucasian woman was admitted to neurology with vertigo, gait ataxia and dysarthria, transient ischaemic attack (TIA) and stroke were quickly excluded. Indicative for the further diagnosis of a paraneoplastic syndrome was the identification of onconeuronal antibodies the detection of which can be associated with certain tumour entities. The strongly positive anti-Yo antibody that is formed above all by breast and lung cancers as well as ovarian cancer led to a corresponding staging. The tumour markers CEA, CA 125 and CA 15-3 were in the normal ranges. Mammography and thorax CT were also unremarkable; on transvaginal sonography the internal genitals were inconspicuous except for a follicular cyst on the left. On abdominal CT the only conspicuous finding was a 1.5 cm ovarian cyst. After consensual agreement a bilateral laparoscopic adenexectomy was performed but with unremarkable abdominal findings. The histological examination confirmed a right-sided undifferentiated tubal carcinoma with the provisional classification FIGO IIA. After a stage-related staging operation, the final classification was found to be the FIGO-IIIC stage on account of positive retroperitoneal lymph nodes. Thus adjuvant chemotherapy with 6 cycles of carboplatin and paclitaxel was performed. By means of a timely, guideline-conform therapy for tubal carcinoma, the neurological symptoms and, above all, the dysarthria could be improved after 10 months.


Atom-efficient route for converting incineration ashes into heavy metal sorbents.
Bottom ashes produced from municipal solid-waste incineration are suitable for sorbent synthesis because of their inherent composition, high alkalinity, metastable mineralogy, and residual heat. This work shows that bottom ashes can be atom-efficiently converted into valuable sorbents without the need for costly and hazardous chemicals. The ashes were hydrothermally treated in rotary autoclaves at autogenic pH conditions to promote the conversion of precursor mineral phases into zeolites and layered silicate hydrates. Two main mineral phases were formed: katoite and sodium aluminum phosphate silicate hydrate. These mineral alterations are accompanied by a tenfold increase in specific surface area and a twofold reduction in average particle size. Performance evaluation of the new sorbents for Cd(2+), Zn(2+), and Pb(2+) adsorption at pH5 indicates sorption capacities of 0.06, 0.08, and 0.22 mmol g(-1), respectively, which are similar to those of natural adsorbents and synthetic materials obtained from more demanding synthesis conditions.


Long-term clinical outcome of living-donor liver transplantation for primary biliary cirrhosis.
We described the recurrence of primary biliary cirrhosis (PBC) after living donor liver transplantation (LDLT) (Liver Transplantation, 7, 2001: 588). However, since the follow-up period in that study was insufficiently long (median 35.5 months), we performed a long-term study to further characterize recurrence of PBC after LDLT. From 1991 to 2006, 15 patients with end-stage PBC underwent LDLT at Tokyo Women's Medical University. Of these patients, we studied 8 PBC patients (age 29 to 51 years, all females) who survived LDLT for more than 5 years. The follow-up period for these patients ranged form 68 to 120 months. Immunosuppression was maintained with tacrolimus and prednisone. Laboratory examinations performed in every patient and donor before LDLT included routine biochemical studies, antimitochondrial antibody (AMA) by immunofluorescence (IF), anti-M2 by enzyme-linked immunosorbent assay as well as antinuclear antibody (ANA) by IF, and immunoglobulin. After LDLT, the same laboratory examinations were performed in patients every 6 months. Liver biopsy was performed when patients exhibited clinical or biochemical signs of graft dysfunction. In addition, protocol biopsy was performed every 1 to 2 years after LDLT. At the time of LDLT, all patients had end-stage cholestatic liver failure. Seven patients were positive for AMAand anti-M2 while 1 patient was negative for these markers but strongly positive for ANA. Donors were blood relatives in 6 cases, and 2 donors who were not blood relatives still exhibited multiple HLA matches with the recipients. At the end of the study in May 2006, all patients were doing well. On laboratory examination, mild abnormal liver function test results were found in 4 patients: 3 were probably due to recurrence of PBC, 1 resulted from nonalcoholic steatohepatitis. Comparison of the AMA titer between before LDLT and the most recent follow-up visit showed an increase in three patients, a decrease in two patients and no change in three patients. In contrast, the ANA titer increased in five patients. Histologically, strong evidence of recurrent PBC was found in 4 patients, and findings compatible with PBC were present in 2 additional patients. Although the number of our patients is small, our findings confirm that PBC can recur at high frequency after LDLT. However, this complication has not developed to advanced stages and has not caused appreciable symptoms in our patients, all of whom have a good quality of life.


Hierarchical Connectome Modes and Critical State Jointly Maximize Human Brain Functional Diversity.
The brain requires diverse segregated and integrated processing to perform normal functions in terms of anatomical structure and self-organized dynamics with critical features, but the fundamental relationships between the complex structural connectome, critical state, and functional diversity remain unknown. Herein, we extend the eigenmode analysis to investigate the joint contribution of hierarchical modular structural organization and critical state to brain functional diversity. We show that the structural modes inherent to the hierarchical modular structural connectome allow a nested functional segregation and integration across multiple spatiotemporal scales. The real brain hierarchical modular organization provides large structural capacity for diverse functional interactions, which are generated by sequentially activating and recruiting the hierarchical connectome modes, and the critical state can best explore the capacity to maximize the functional diversity. Our results reveal structural and dynamical mechanisms that jointly support a balanced segregated and integrated brain processing with diverse functional interactions, and they also shed light on dysfunctional segregation and integration in neurodegenerative diseases and neuropsychiatric disorders.


Lactobacillus herbarum sp. nov., a species related to Lactobacillus plantarum.
Strain TCF032-E4 was isolated from a traditional Chinese fermented radish. It shares >99% 16S rRNA sequence identity with L. plantarum, L. pentosus and L. paraplantarum. This strain can ferment ribose, galactose, glucose, fructose, mannose, mannitol, N-acetylglucosamine, amygdalin, arbutin, salicin, cellobiose, maltose, lactose, melibiose, trehalose and gentiobiose. It cannot ferment sucrose, which can be used by L. pentosus, L. paraplantarum, L. fabifermentans, L. xiangfangensis and L. mudanjiangensis, as well as most of the L. plantarum strains (88.7%). TCF032-E4 cannot grow at temperature above 32 °C. This strain shares 78.2-83.6% pheS (phenylalanyl-tRNA synthetase alpha subunit) and 89.5-94.9% rpoA (RNA polymerase alpha subunit) sequence identity with L. plantarum, L. pentosus, L. paraplantarum, L. fabifermentans, L. xiangfangensis and L. mudanjiangensis. These results indicate that TCF032-E4 represents a distinct species. This hypothesis was further confirmed by whole-genome sequencing and comparison with available genomes of related species. The draft genome size of TCF032-E4 is approximately 2.9 Mb, with a DNA G+C content of 43.5 mol%. The average nucleotide identity (ANI) between TCF032-E4 and related species ranges from 79.0 to 81.1%, the highest ANI value being observed with L. plantarum subsp. plantarum ATCC 14917T. A novel species, Lactobacillus herbarum sp. nov., is proposed with TCF032-E4T ( = CCTCC AB2015090T = DSM 100358T) as the type strain.


Disorders in high-density metabolism with insulin resistance and chronic kidney disease.
Cardiovascular risk increases with each decrement in renal function. Low-density lipoprotein (LDL) cholesterol levels are not associated with increased mortality, but high-density lipoprotein (HDL) levels are inversely associated with cardiovascular risk. Lipoprotein composition with increased abundance of small dense LDL and HDL and reduced levels of more buoyant isoforms is similar to what is found in states of insulin resistance and in the metabolic syndrome (MS). In both cases, high triglyceride levels are associated with reduced HDL levels. Chronic kidney disease (CKD) is itself associated with increasing insulin resistance as renal function fails. In both instances, decreased levels of apo A-I and apo A-II are a consequence of increased fractional catabolic rate (FCR), resulting from a predominance of small HDL particles. HDL maturation is impaired in CKD through decreased activity of lecithin:cholesterol acyltransferase (LCAT), and increased cholesterol ester transfer protein (CETP) activity in MS shuttles triglycerides back into HDL, thereby destabilizing it. Whether insulin resistance is entirely responsible for disorders of HDL metabolism in CKD, or whether the process is a result of unrelated pathophysiology, is currently unknown.


Differential activation-dependent regulation of integrin function in cultured human T-leukemic cell lines.
T lymphocytes isolated from human peripheral blood express beta 1 (VLA) and LFA-1 integrins, but strong binding to integrin ligands occurs only after the delivery of an activation stimulus to the T cell. To gain further insight into activation-dependent regulation of integrin function, we have analyzed integrin activity on three different T-leukemic cell lines: Jurkat, CEM, and H9. This analysis shows important mechanistic differences in integrin regulation. First, phorbol ester treatment results in increased beta 1 integrin-dependent adhesion of both Jurkat and CEM cells to fibronectin, but decreased adhesion of H9 cells. Second, certain activation stimuli that upregulate beta 1 integrin activity in peripheral T cells are nonfunctional in these T-cell lines. Third, analysis of a panel of Jurkat mutants lacking surface expression of CD2 and/or CD3 shows that CD2-mediated upregulation of beta 1 integrin activity is dependent on expression of CD3, whereas CD28-mediated upregulation is not dependent on either CD2 or CD3 expression. Fourth, all T-cell lines tested show an inability to adhere to purified ICAM-1 via LFA-1. The selective alterations in integrin regulation in these cell lines relative to peripheral blood T cells provide important insights into the intracellular processes involved in integrin activation.


Delivery and financing of oral health care in long-term services and supports: A scoping review.
The oral health of older adults requiring long-term services and supports is reported to be poor as there is no national standard of care for the provision of oral health care. The purpose of this scoping review was to understand the breadth of models of delivery and financing of oral health care in the full spectrum of long-term services and supports. A literature search was performed in 4 electronic databases: MEDLINE via PubMed interface, Embase, Cumulative Index to Nursing and Allied Health Literature, and AgeLine. Included articles were those that were regarding a nursing home population or dependent older adults living in the community, included a delivery or financing model for oral health care, and included an outcome measurement. Sixteen articles were included in the review. Delivery mechanisms included onsite mobile oral health care at nursing homes and adult day health care centers for those living in the community or home visits for those who were homebound. Other mechanisms included teledentistry or using alternative workforce models such as certified public health dental hygienists. Numerous studies reported positive oral health outcomes when comprehensive care was provided in a variety of settings. Other reported outcomes included oral health stability, caries indexes, cost, and oral health-related quality of life. If providing onsite oral health care is not possible at facilities, programs can consider home visits, teledentistry, and alternative workforce models.


Increased IL-17A secretion in response to Candida albicans in autoimmune polyendocrine syndrome type 1 and its animal model.
Autoimmune polyendocrine syndrome type 1 (APS-1) is a multiorgan autoimmune disease caused by mutations in the autoimmune regulator (AIRE) gene. Chronic mucocutaneous candidiasis, hypoparathyroidism and adrenal failure are hallmarks of the disease. The critical mechanisms causing chronic mucocutaneous candidiasis in APS-1 patients have not been identified although autoantibodies to cytokines are implicated in the pathogenesis. To investigate whether the Th reactivity to Candida albicans (C. albicans) and other stimuli was altered, we isolated PBMC from APS-1 patients and matched healthy controls. The Th17 pathway was upregulated in response to C. albicans in APS-1 patients, whereas the IL-22 secretion was reduced. Autoantibodies against IL-22, IL-17A and IL-17F were detected in sera from APS-1 patients by immunoprecipitation. In addition, Aire-deficient (Aire(0/0) ) mice were much more susceptible than Aire(+/+) mice to mucosal candidiasis and C. albicans-induced Th17- and Th1-cell responses were increased in Aire(0/0) mice. Thus an excessive IL-17A reactivity towards C. albicans was observed in APS-1 patients and Aire(0/0) mice.


Estrogen-nucleic acid adducts: guanine is major site for interaction between 3,4-estrone quinone and COIII gene.
The carcinogenicity of estrogens in rodents and man has been attributed to either alkylation of cellular macromolecules and/or redox-cycling, generation of active radicals and DNA damage. Metabolic activation of estradiol leading to the formation of catechol estrogens is believed to be a prerequisite for its genotoxic effects. 4-Hydroxyestradiol is a potent inducer of tumors in hamsters. Previous studies have shown that 3,4-estrone quinone (3,4-EQ) can redox-cycle and is capable of inducing exclusively single strand DNA breaks in MCF-7 breast cancer cells, as well as react with various nucleophiles including amino acids and nucleic acids to give Michael addition products. In this paper we examined the nature of the interaction of 3,4-EQ with COIII gene and analysed the estrogen-DNA adducts by 32P-post-labeling. The reaction of 3,4-EQ with the COIII gene followed by polymerase arrest assay showed several stop sites in which guanine was preferentially attacked by 3,4-EQ and, to a lesser extent, with Ade, Cyt and Thy. 32P-Post-labeling analysis of the reaction of 3,4-EQ with COIII gene gave one major adduct which was found to be identical to that obtained from reaction of dGMP with 3,4-EQ. The observation that obstruction of in vitro replication of COIII template bound to 3,4-EQ suggests that estrogen quinone adducted lesions can arrest DNA polymerase. These results indicate that 3,4-EQ may be genotoxic and may provide one possible explanation for the carcinogenic effects of estrogens.


A Gut Symbiotic Bacterium Serratia marcescens Renders Mosquito Resistance to Plasmodium Infection Through Activation of Mosquito Immune Responses.
The malaria development in the mosquito midgut is a complex process that results in considerable parasite losses. The mosquito gut microbiota influences the outcome of pathogen infection in mosquitoes, but the underlying mechanisms through which gut symbiotic bacteria affect vector competence remain elusive. Here, we identified two Serratia strains (Y1 and J1) isolated from field-caught female Anopheles sinensis from China and assessed their effect on Plasmodium development in An. stephensi. Colonization of An. stephensi midgut by Serratia Y1 significantly renders the mosquito resistant to Plasmodium berghei infection, while Serratia J1 has no impact on parasite development. Parasite inhibition by Serratia Y1 is induced by the activation of the mosquito immune system. Genome-wide transcriptomic analysis by RNA-seq shows a similar pattern of midgut gene expression in response to Serratia Y1 and J1 in sugar-fed mosquitoes. However, 24 h after blood ingestion, Serratia Y1 modulates more midgut genes than Serratia J1 including the c-type lectins (CTLs), CLIP serine proteases and other immune effectors. Furthermore, silencing of several Serratia Y1-induced anti-Plasmodium factors like the thioester-containing protein 1 (TEP1), fibrinogen immunolectin 9 (FBN9) or leucine-rich repeat protein LRRD7 can rescue parasite oocyst development in the presence of Serratia Y1, suggesting that these factors modulate the Serratia Y1-mediated anti-Plasmodium effect. This study enhances our understanding of how gut bacteria influence mosquito-Plasmodium interactions.


Glomus tumor masquerading for 22 years as osteoarthritis of the hip.
Glomus tumors are rare benign mesenchymal neoplasms that account for less than 2% of soft tissue tumors. These neoplasms typically are small nodules less than 1 cm in diameter, associated with pain that is exacerbated by tactile stimulation and cold hypersensitivity. We present a case of a large glomus tumor of the left lateral hip associated with a long history of severe pain of the left hip interfering with ambulation. Chronic pain as a result of a subcutaneous glomus tumor is rare and frequently misdiagnosed. In the case reported, a solid glomus tumor presented with 22 years of unilateral hip pain attributed to posttraumatic degenerative joint disease. Excision of a 4 x 3-cm nodule resulted in complete resolution of tenderness and joint pain. Subcutaneous glomus tumors can have unusually large size and location and should be considered in the differential diagnosis of chronic, atypical, or treatment-resistant joint pain.


TGF-beta(1) downregulates PTHrP in coronary endothelial cells.
Parathyroid hormone-related peptide (PTHrP) is expressed throughout the cardiovascular system including coronary endothelial cells. Factors involved in the regulation of cardiac PTHrP expression have not been examined before. This study investigates the influence of transforming growth factor (TGF)-beta(1)on ventricular PTHrP expression. Coronary endothelial cells were isolated from ventricles of adult rats and PTHrP protein expression in these cultures was analysed by immunoblotting. TGF-beta(1)caused a concentration-dependent reduction in PTHrP protein within 24 h. In transgenic mice over-expressing TGF-beta(1)ventricular PTHrP protein expression and release was reduced compared to non-transgenic littermates. Similar concerns hold for PTHrP mRNA content (RT-PCR). Since ventricular TGF-beta(1)expression increases under pathophysiological conditions like arterial hypertension, ventricular PTHrP expression was further determined in aging spontaneously hypertensive (SHR-SP) and normotensive rats. TGF- beta(1)expression was increased in SHR-SP and ventricular PTHrP mRNA expression was downregulated at the age of 10 months. PTHrP expression did not recover in elder SHR-SP in which TGF-beta(1)expression was normalized again. Finally, we investigated ventricular PTHrP expression in rats after banding of the ascending aorta which generates a pressure induced hypertrophy without an induction of TGF-beta(1)expression. In ventricles from these animals, PTHrP expression was transiently increased and normalized at day 3. In conclusion, PTHrP expression was reduced under all conditions in which coronary endothelial cells were exposed to TGF-beta(1). PTHrP expression does not correlate with cardiac hypertrophy. Since coronary endothelial cells represent the majority of PTHrP producing cells in the ventricle its downregulation by TGF- beta(1)seems to be relevant for the paracrine effects of PTHrP.


Echocardiographic guidance and monitoring of left atrial appendage closure with AtriClip during open-chest cardiac surgery.
Left atrial appendage (LAA) closure prevents thromboembolic risk and avoids lifelong anticoagulation due to atrial fibrillation (AF). Nowadays, AtriClip, a modern epicardial device approved in June 2010, allows external and safe closure of LAA in patients undergoing cardiac surgery during other open-chest cardiac surgical procedures. Such a surgical approach and its epicardial deployment differentiates LAA closure with AtriClip from percutaneous closure techniques such as Watchman (Boston Scientific, Marlborough, MA, USA), Lariat (SentreHEART Inc., Redwood City, CA, USA), and Amplatzer Amulet (St. Jude Medical, St. Paul, MN, USA) device procedures. AtriClip positioning must consider perioperative transesophageal echocardiography (TEE) to confirm LAA anatomical features, to explore the links with neighboring structures, and finally to assess its successful closure. We report a sequence of images to document the role of intraoperative TEE during an elective aortic valve replacement and LAA external closure with AtriClip.


How does colistin-induced nephropathy develop and can it be treated?
Colistin is an old antibiotic used in the treatment of Gram-negative infections. It was once suspended because of its nephrotoxic effect but has since been reintroduced due to multidrug-resistant bacterial infections. The pathogenesis of colistin-associated nephropathy has not been clarified, and there is currently no effective therapeutic or prophylactic agent available. The aim of this study was to investigate the roles of caspase-associated apoptosis and caspase 1, calpain 1, inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS) expression in the pathogenesis of colistin-associated nephrotoxicity and the effect of grape seed proanthocyanidin extract (GSPE) in preventing it. Twenty-four rats were divided into three groups: control, colistin, and colistin plus GSPE (colistin+GSPE). Colistin-associated nephropathy was induced by the administration of 300,000 IU/kg of body weight/day colistin intraperitoneally for 7 days. The experiment was discontinued on the seventh day. Blood was collected for measurements of blood urea nitrogen (BUN) and creatinine levels. Histopathological examination of kidney tissue and caspase 1 and 3, iNOS, eNOS, terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL), and calpain 1 staining was also performed. Significant increases in BUN levels; creatinine levels; renal histopathological scores; and TUNEL, caspase 1 and 3, calpain 1, iNOS, and eNOS staining were observed for the colistin group compared to the control group. Significant decreases in BUN levels; creatinine levels; renal histopathological scores; and TUNEL, caspase 1 and 3, calpain 1, iNOS, and eNOS staining were observed in the colistin+GSPE group compared to the colistin group. Our study shows, for the first time in the literature, that caspase-mediated apoptosis, iNOS, caspase 1, and calpain 1 are involved in the pathogenesis of colistin-associated nephropathy. GSPE had a renoprotective effect, as shown by the lowered levels of these mediators.


An improved automated method for the measurement of thermal thresholds. 2. Patients with peripheral neuropathy.
Thermal thresholds were determined by a new technique, at wrists and ankles in 143 patients with peripheral neuropathies of diverse aetiologies. Ninety-nine percent of patients (141/143) had abnormalities of one or both thresholds. In only two patients with mild/early Friedreich's ataxia were thermal thresholds normal. Electromyography was performed and fastest motor nerve conduction velocities and sensory nerve action potential parameters were measured in all the patients using conventional techniques in ulnar, median and sural nerves. Eighty-nine percent of patients (127/143) had one or more abnormalities on these electrophysiological studies. However, 39 of 40 patients with completely normal sensory nerve studies had an abnormality of one or more thermal thresholds. Eighty-six percent of 48 patients with normal sural nerve studies had abnormal thermal thresholds at the ankle. Sixty percent of 70 patients with normal sensory median and ulnar nerve studies had abnormal wrist thermal thresholds. This improved technique for the determination of thermal thresholds reveals that disturbances of thermal sensibility are present in the majority of peripheral neuropathies irrespective of aetiology. In some patients disturbances of thermal thresholds antedate the appearance of abnormalities on conventional electrophysiological investigation. The findings suggest that this technique has considerable usefulness in the detection of small nerve fibre dysfunction in the context of generalised neuropathy.


A phase I study of the combination of temsirolimus with irinotecan for metastatic sarcoma.
mTOR inhibitors are emerging as important anti-neoplastic agents with a wide range of clinical applications. The topoisomerase I inhibitor irinotecan is a potent DNA damaging drug, with a broad spectrum of anticancer activities. mTOR appears to enhance cancer cell survival following DNA damage, thus the inhibition of mTOR after irinotecan could theoretically show synergistic activities in patients. Both mTOR inhibitors and irinotecan have been used as single agents in soft tissue sarcomas with limited efficacy. We completed a phase I trial of the combination of the mTOR inhibitor, temsirolimus, and irinotecan in patients with advanced soft tissue sarcoma. Seventeen patients were recruited. The Phase II recommended dose is 20 mg of temsirolimus and 80 mg/m2 of irinotecan administered on weekly basis for three out of four weeks. Most frequently encountered toxicities include cytopenias, fatigue, and gastrointestinal toxicities. Two patients (one with leiomyosarcoma and one with high grade undifferentiated sarcoma) had stable disease for more than 12 months.


Renal cytochrome P-450-arachidonic acid metabolism: localization and hormonal regulation in SHR.
Epoxygenase and omega- and omega-1-hydroxylases are the major cytochrome P-450-arachidonate (P-450-AA) metabolizing enzymes in renal tissues. We measured P-450-AA metabolism in single nephron segments and determined the tubular localization of this activity in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). Formation of 20-hydroxyeicosatetraenoic acid (20-HETE), the product of AA omega-hydroxylase was specifically localized in the entire proximal tubules (S1, S2, and S3 segments), whereas formation of 19-HETE, the product of omega-1-hydroxylase and epoxyeicosatrienoic acids (EETs), products of AA epoxygenase, was demonstrable throughout the tubule. Although distribution patterns were similar in SHR and WKY, formation of 19- and 20-HETE in the proximal tubules was higher in SHR, whereas the formation of EETs was not different between the two strains. In the proximal tubules, angiotensin II (ANG II) significantly stimulated epoxygenase activity (EETs formation), whereas parathyroid hormone (PTH) and epidermal growth factor (EGF) had no effect on epoxygenase but significantly stimulated omega-hydroxylase activity (20-HETE formation). Because P-450-AA metabolites have a wide and contrasting spectrum of biological and renal effects, from vasodilation to vasoconstriction and from inhibition to stimulation of Na(+)-K(+)-adenosinetriphosphatase, their localization to the specific nephron segments and differential stimulation of their formation by ANG II, PTH, and EGF may contribute not only to renal hemodynamics and blood pressure regulation but also to the regulation of renal sodium and water balance.


Mitochondrial DNA haplogroup K is associated with a lower risk of Parkinson's disease in Italians.
It has been proposed that European mitochondrial DNA (mtDNA) haplogroups J and K, and their shared 10398G single-nucleotide polymorphism (SNP) in the ND3 gene, are protective from Parkinson's disease (PD). We evaluated the distribution of the different mtDNA haplogroups in a large cohort of 620 Italian patients with adult-onset (>50, <65 years of age) idiopathic PD vs two groups of ethnic-matched controls. Neither the frequencies of haplogroup J nor that of 10398G were significantly different. However, the frequency of haplogroup K was significantly lower in PD. Stratification by sex and age indicated that the difference in the distribution of haplogroup K was more prominent in >50 year old males. In spite of the common 10398G SNP, haplogroups J and K belong to widely diverging mitochondrial clades, a consideration that may explain the different results obtained for the two haplogroups in our cohorts. Our study suggests that haplogroup K might confer a lower risk for PD in Italians, corroborating the idea that the mitochondrial oxidative phosphorylation pathway is involved in the susceptibility to idiopathic PD.


[Effect of chorionic gonadotropin on the structure and endocrine function of the thymus gland in mice].
The thymic secretory function changes in the pubertal female mice under the influence of chorionic gonadotrophin (CG). The latter is activated after administration of hormone in doses, corresponding to its levels in animals in different terms of pregnancy and is depressed with doses, greatly exceeding those levels. The lack of the effect in thymectomized mice testifies to the realization of the modulating CG action at the thymus level. Changes of mass and cellularity of the thymus and spleen reflect dose-dependent alteration of the thymocyte migration rate.


Antiviral Activity of HIV gp120-Targeting Bispecific T Cell Engager Antibody Constructs.
Today's gold standard in HIV therapy is combined antiretroviral therapy (cART). It requires strict adherence by patients and lifelong medication, which can lower the viral load below detection limits and prevent HIV-associated immunodeficiency but cannot cure patients. The bispecific T cell-engaging (BiTE) antibody technology has demonstrated long-term relapse-free outcomes in patients with relapsed and refractory acute lymphocytic leukemia. Here, we generated BiTE antibody constructs that target the HIV-1 envelope protein gp120 (HIV gp120) using either the scFv B12 or VRC01, the first two extracellular domains (1 + 2) of human CD4 alone or joined to the single chain variable fragment (scFv) of the antibody 17b fused to an anti-human CD3ε scFv. These engineered human BiTE antibody constructs showed engagement of T cells for redirected lysis of HIV gp120-transfected CHO cells. Furthermore, they substantially inhibited HIV-1 replication in peripheral blood mononuclear cells (PBMCs) as well as in macrophages cocultured with autologous CD8+ T cells, the most potent being the human CD4(1 + 2) BiTE [termed CD(1 + 2) h BiTE] antibody construct and the CD4(1 + 2)L17b BiTE antibody construct. The CD4(1 + 2) h BiTE antibody construct promoted HIV infection of human CD4-/CD8+ T cells. In contrast, the neutralizing B12 and the VRC01 BiTE antibody constructs, as well as the CD4(1 + 2)L17b BiTE antibody construct, did not. Thus, BiTE antibody constructs targeting HIV gp120 are very promising for constraining HIV and warrant further development as novel antiviral therapy with curative potential.IMPORTANCE HIV is a chronic infection well controlled with the current cART. However, we lack a cure for HIV, and the HIV pandemic goes on. Here, we showed in vitro and ex vivo that a BiTE antibody construct targeting HIV gp120 resulted in substantially reduced HIV replication. In addition, these BiTE antibody constructs display efficient killing of gp120-expressing cells and inhibited replication in ex vivo HIV-infected PBMCs or macrophages. We believe that BiTE antibody constructs recognizing HIV gp120 could be a very valuable strategy for a cure of HIV in combination with cART and compounds which reverse latency.


Can radiographic patellofemoral osteoarthritis be diagnosed using clinical assessments?
The aim of this study was to determine whether participant characteristics and clinical assessments could identify radiographic osteoarthritis (OA) in individuals with clinically diagnosed, symptomatic patellofemoral osteoarthritis (PFOA). Participant characteristics and clinical assessments were obtained from 179 individuals aged 50 years and over with clinically diagnosed symptomatic PFOA, who were enrolled in a randomised trial. Anteroposterior, lateral, and skyline X-rays were taken of the symptomatic knee. The presence of radiographic PFOA was defined as "no or early PFOA" (Kellgren and Lawrence [KL] grade ≤1 in the PF compartment) or "definite PFOA" (KL grade ≥2). Diagnostic test statistics were applied to ascertain which participant characteristics and clinical assessments could identify the presence of definite radiographic PFOA. A total of 118 participants (66%) had definite radiographic PFOA. Univariate analysis identified that older age (>61 years), female sex, higher body mass index (BMI) (>29 kg/m2 ), longer pain duration (>2.75 years), higher maximum knee pain during stair ambulation (>47/100 mm), and fewer repeated single step-ups to pain onset (<21) were associated with the presence of definite radiographic PFOA. Multivariate logistic regression indicated that BMI, pain duration, and repeated single step-ups to pain onset were independently associated with radiographic PFOA and identified the presence of definite radiographic PFOA with an overall accuracy of 73%. In individuals over 50 years of age with a clinical diagnosis of PFOA, higher BMI, longer pain duration, and fewer repeated single step-ups to pain onset increased the likelihood of radiographic PFOA. However, overall diagnostic accuracy was modest, suggesting that radiographic PFOA cannot be confidently identified using these tests.


Functional validation of putative toxin-antitoxin genes from the Gram-positive pathogen Streptococcus pneumoniae: phd-doc is the fourth bona-fide operon.
Bacterial toxin-antitoxin (TAs) loci usually consist of two genes organized as an operon, where their products are bound together and inert under normal conditions. However, under stressful circumstances the antitoxin, which is more labile, will be degraded more rapidly, thereby unleashing its cognate toxin to act on the cell. This, in turn, causes cell stasis or cell death, depending on the type of TAs and/or time of toxin exposure. Previously based on in silico analyses, we proposed that Streptococcus pneumoniae, a pathogenic Gram-positive bacterium, may harbor between 4 and 10 putative TA loci depending on the strains. Here we have chosen the pneumococcal strain Hungary(19A)-6 which contains all possible 10 TA loci. In addition to the three well-characterized operons, namely relBE2, yefM-yoeB, and pezAT, we show here the functionality of a fourth operon that encodes the pneumococcal equivalent of the phd-doc TA. Transcriptional fusions with gene encoding Green Fluorescent Protein showed that the promoter was slightly repressed by the Phd antitoxin, and exhibited almost background values when both Phd-Doc were expressed together. These findings demonstrate that phd-doc shows the negative self-regulatory features typical for an authentic TA. Further, we also show that the previously proposed TAs XreA-Ant and Bro-XreB, although they exhibit a genetic organization resembling those of typical TAs, did not appear to confer a functional behavior corresponding to bona fide TAs. In addition, we have also discovered new interesting bioinformatics results for the known pneumococcal TAs RelBE2 and PezAT. A global analysis of the four identified toxins-antitoxins in the pneumococcal genomes (PezAT, RelBE2, YefM-YoeB, and Phd-Doc) showed that RelBE2 and Phd-Doc are the most conserved ones. Further, there was good correlation among TA types, clonal complexes and sequence types in the 48 pneumococcal strains analyzed.


Specific beta-adrenergic mechanisms in the hypoglycaemic activation of gastrin and gastric acid secretion.
To study whether specific beta-adrenergic mechanisms contribute to the hypoglycaemic activation of gastrin and gastric acid secretion, the effects of racemic and dextroisomer propranolol (0.1 mg/kg, intravenously) were studied during insulin tests (0.2 IU/kg) in 13 persons. dl-Propranolol inhibited the gastrin response to hypoglycaemia markedly and more than the insignificant alteration observed after d-propranolol. Gastric acid response to hypoglycaemia was significantly reduced by dl-propranolol and not by d-propranolol. The findings demonstrate that non-beta-adrenergic effects of propranolol on the stomach are minor and that specific beta-adrenergic mechanisms are directly or indirectly involved in the hypoglycaemic stimulation of the stomach.


Appendiceal mucocele. A case report and literature review.
Appendiceal mucocele is a rare disease (0.3% of all appendectomy) and is characterized by the accumulation of mucoid material in the appendiceal lumen. Etiopathogenesis can be inflammatory or neoplastic. Four entities can be distinguished on the basis of histopathologic epithelial characteristics: simple appendiceal mucocele (AM), mucocele with epithelial hyperplasia, cystadenoma and cystadenocarcinoma; the last two subgroups represent neoplastic forms. Dissemination of neoplastic cells and mucoid material in abdominal cavity, caused by appendiceal perforation, clinically results in pseudomyxoma peritonei which is the dramatic evolution in 10-15% of cases. Clinically it can remain either asymptomatic for long time or it can manifest with abdominal pain that can be associated with the presence of a palpable mass. The most common clinical manifestation is pain in the right iliac fossa. Preoperative diagnosis is rare, while it is more frequently intraoperative. Therapy is fundamentally surgical: appendectomy is curative for simple AM, for AM with epithelial hyperplasia and for cystadenoma with intact appendiceal base; cecum resection is indicated for cystadenoma with larger base of implantation; right hemicolectomy has been the elective treatment in case of cystadenocarcinoma for several years although Gonzalez-Moreno and Sugarbaker have recently demonstrated its validity as definitive treatment only if it is performed in order to obtain complete cytoreduction, if there is lymph node involvement, or if histopathological examination indicates non-mucinous type. We report the case of a 60-year-old woman that presented with cystic neoformation in the right iliac fossa, that was preoperatively considered deriving from the ovary. We intraoperatively found the presence of appendiceal mucocele that histological examination defined as mucinous cystadenoma.


[Serine proteases from Bac. subtilis].
Using biospecific adsorbent and subsequent gel-filtration of Sephadex G-75 three fractions of serine proteases (I--III) having different physicochemical properties were isolated from Bac. subtilis. The first protease had molecular weight of 23000--24000 (pH optimum 6,5, activation energy 16,6 ccal/mol. The second one had molecular weight of 29000, pH optimum 11,0, activation energy 14,4 ccal/mol. The third protease was a mixture of proteases with average molecular weights 26000 and pH optima at 7,0, 8,5 and 11,0.


[Structuro-functional properties of the bacteria Bacillus brevis in relation to the accumulation of gramicidin S in cells].
The culture of Bacillus brevis var. G-B R-form was grown in the presence of beta-phenyl-beta-alanine, the inhibitor of gramicidin S synthesis, is characterized by enhanced endogenous respiration and the DPI-reductase activity as compared to the culture synthezising antibiotic. The increased synthesis of the antibiotic in the region of the culture transition from the logarithmic growth phase to the linear one is associated with a decrease in the number of viable cells despite the fact that the culture on the whole does not die but continues to grow. The membranes prepared from young gramicidin S-free cells and from the cells enriched with the antibiotic possess identical electron micrograph images, IR spectra and protein sets as determined by polyacrylamide gel electrophoresis in a Na-DS system. However, in young cell membranes NADH and succinate dehydrogenase are insensitive to gramicidin S and only malate dehydrogenase is inhibited by this antibiotic. In aged cell membranes the activities of all mentioned dehydrogenases are suppressed. Malate dehydrogenase from young cells is weakly inhibited by thyrotrycin obtained from Bac. brevis ATCC 10068; succinate dehydrogenase is entirely insensitive to this antibiotic, while NADH-dehydrogenase is almost completely inhibited by it. The specificity of action on the respiratory chain of peptide antibiotics synthesized by the cells of one strain of Bac. brevis is suggestive of a possible regulatory role of these peptides in the metabolism of the producent. Hence the accumulation of gramicidin S which is adsorbed on the membrane and destroys the respiratory chain function to the cause of the low rate of oxygen uptake by the culture of Bac. brevis var. G-B R-form and of the low activities of DPI-reductases.


Influence of postoperative hepatic angiography on mortality after laparotomy in Grade IV/V hepatic injuries.
Mortality rate for severe liver injuries remains high. As an adjunct to surgery, postoperative hepatic angiography (PHA) may have a positive impact on outcomes. This study sought to compare outcomes following surgical management of severe liver injuries with and without PHA using propensity score matching analysis. Data from the National Trauma Data Bank from 2007 to 2014 were analyzed. The study population consisted of patients older than 18 years, sustaining severe liver injuries (i.e., American Association for the Surgery of Trauma Organ Injury Scale (AAST-OIS) Grade IV or V) who underwent surgery. Patients were divided into two groups. The PHA group consisted of those undergoing surgery followed by PHA. In the surgery-only group, no angiography was performed. To determine the impact of PHA on outcomes, propensity score matching analysis (1:3) was used. A total of 3,871 patients met inclusion criteria. Of those, 205 (5.3%) patients underwent PHA. Prior to matching, patients in the PHA group had higher severity, but overall in-hospital mortality was found to be similar between the two groups. After 1:3 propensity-score matching, 196 patients in the PHA group were matched with 588 in the surgery-only group with well-balanced baseline characteristics. The in-hospital mortality was significantly lower in the PHA group compared with the surgery-only group (24.5% vs. 35.9%; odds ratio, 0.58; 95% confidence interval, 0.40-0.84). However, hospital length of stay was longer (16.0 [7.0-29.8] vs. 11 [1.0-25.0] days, p = 0.001), and the incidence of deep and organ/space surgical site infection (3.6% vs. 1.2%, 8.2% vs. 3.5%, respectively) was higher in the PHA group. The use of PHA was associated with decreased mortality rates. A multimodality approach using both surgical intervention followed by PHA appears to identify patients that may benefit from arterial embolization, leading to decreased mortality of severe liver injuries. Therapeutic study, level IV.


Deeply sequenced metagenome and metatranscriptome of a biogas-producing microbial community from an agricultural production-scale biogas plant.
The production of biogas takes place under anaerobic conditions and involves microbial decomposition of organic matter. Most of the participating microbes are still unknown and non-cultivable. Accordingly, shotgun metagenome sequencing currently is the method of choice to obtain insights into community composition and the genetic repertoire. Here, we report on the deeply sequenced metagenome and metatranscriptome of a complex biogas-producing microbial community from an agricultural production-scale biogas plant. We assembled the metagenome and, as an example application, show that we reconstructed most genes involved in the methane metabolism, a key pathway involving methanogenesis performed by methanogenic Archaea. This result indicates that there is sufficient sequencing coverage for most downstream analyses. Sequenced at least one order of magnitude deeper than previous studies, our metagenome data will enable new insights into community composition and the genetic potential of important community members. Moreover, mapping of transcripts to reconstructed genome sequences will enable the identification of active metabolic pathways in target organisms.


Building Capacity for Caregiver Education in Yap, Micronesia.
The US Affiliated Pacific Islands have an urgent need for family caregiver education to prevent caregiver burnout and strengthen the existing culture where seniors are cared for at home by their families. The Pacific Islands Geriatric Education Center conducted a 32-hour family caregiver train-thetrainer workshop in partnership with the Yap Department of Health Services and the Yap Area Health Education Center (AHEC) from October 16 - 20, 2017. Twenty-seven participants including community health workers, peer educators, health assistants, nurses, and physicians were trained as instructors. Confidence in caregiving increased following the training and feedback was extremely positive. Competence in geriatric syndromes was improved after attending the workshop (P < .001). Lessons from the field revealed an immense value of adding home visits to the training practicum as well as the need to translate caregiving handouts into the outer island languages. Yap AHEC is committed to offering this course as part of caregiver education at the hospital and in the community.


Hematopoietic stem cells from poor and good mobilizers are qualitatively equivalent.
Marrow damage from chemo- and radiation therapies has been suggested to affect quality and quantity of hematopoietic stem cell (HSC) products. We tested the hypothesis that CD34+ cells (HSCs) from low mobilizers are qualitatively inferior to HSCs from high mobilizers. HSC quality was defined by proportion of primitive HSC subsets (CD34+CD38-, CD34+HLA-DR-, and CD34+ in G0 stage of cell cycle), the proportion of HSCs that express CXCR4 and CD26 homing proteins, and days to neutrophil and platelet (PLT) engraftments after transplant. HSC content and CD34 subsets analyses were performed using flow cytometry following the ISHAGE protocol. We evaluated the HSC quantity and quality of 139 autologous filgrastim-mobilized HSC products. Patients were categorized into low, moderate, and high mobilizers if their total HSC collection was less than 3 × 10(6), 3 × 10(6) or more and less than 5 × 10(6), and 5 × 10(6)/kg or more, respectively. The median number of primitive CD34 subsets increases with increasing HSC numbers and this association was significant (p = 0.001). However, when the ratios of the primitive CD34 subsets to total HSC counts were compared among the mobilization groups, the ratios were not significantly different. Coexpression of neither CD26 nor CXCR4 with CD34 antigen correlated with HSC mobilization. Evaluation of days to neutrophil engraftment among the mobilization groups did not show a significant difference (p = 0.1). However, days to PLT engraftment among the mobilization groups was significantly different (p = 0.05). The quality of HSCs from low mobilizers was comparable to HSCs from high mobilizers.


Bioactive peptide isolated from casein phosphopeptides promotes calcium uptake in vitro and in vivo.
Casein phosphopeptides (CPPs) have been demonstrated to be calcium chelators. Unfortunately, few studies have been reported on the effects of CPPs on the mechanism of the uptake and absorption of Ca2+ and bone metabolism. In this study, a monomeric peptide fraction isolated by RP-HPLC (F6-1) that possessed high calcium transport capacity in Caco-2 cell monolayers was separated and characterized. The effects of F6-1 on the absorption mechanisms of Ca2+ in a Caco-2 monolayer model and bone metabolism in rats were investigated. F6-1 was isolated by preparative and analytical RP-HPLC. Results for calcium transport suggested that the rates of Ca2+ transportation by F6-1 were approximately 2.57, 2.87 and 2.38 times higher than those in the control group at 30, 60 and 120 min, respectively. Results of ultraviolet (UV) spectroscopy indicated that the intensity of UV absorption changed because of the binding of Ca2+ to F6-1. Analysis of transepithelial electrical resistance (TEER) and the expression of TRPV6 in Caco-2 cells showed that F6-1 was likely to influence the transcellular pathway of intestinal absorption of Ca2+ rather than the paracellular pathway. Furthermore, the F6-1 group (1% Ca, 0.03% F6-1) exhibited increases in serum Ca2+ levels, femur length and femur Ca and decreases in serum alkaline phosphatase (ALP) levels and urinary pyridinoline content in a Sprague-Dawley rat model, which implied that F6-1 was beneficial for bone calcification. Overall, our results suggested that F6-1 enhanced the transport of Ca2+ in Caco-2 cells by affecting the transcellular pathway by upregulating the expression of TRPV6. F6-1 also improved bone formation and prevented bone resorption to benefit bone health in rats, which provided a basis for using F6-1 in calcium supplements or functional foods.


Dietary Pattern and Plasma BCAA-Variations in Healthy Men and Women-Results from the KarMeN Study.
Branched-chain amino acids (BCAA) in plasma are discussed as risk factors for the onset of several diseases. Information about the contribution of the overall diet to plasma BCAA concentrations is controversial. Our objective was to investigate which dietary pattern is associated with plasma BCAA concentrations and whether other additional nutrients besides BCAA further characterize this dietary pattern. Based on the cross-sectional KarMeN study, fasting plasma amino acid (AA) concentrations, as well as current and habitual dietary intake were assessed in 298 healthy individuals. Using reduced rank regression, we derived a habitual dietary pattern that explained 32.5% of plasma BCAA variation. This pattern was high in meat, sausages, sauces, eggs, and ice cream but low in nuts, cereals, mushrooms, and pulses. The age, sex, and energy intake adjusted dietary pattern score was associated with an increase in animal-based protein together with a decrease in plant-based protein, dietary fiber, and an unfavorable fatty acid composition. Besides BCAA, alanine, lysine and the aromatic AA were positively associated with the dietary pattern score as well. All of these factors were reported to be associated with risk of type 2 diabetes and cardiovascular diseases before. Our data suggest that rather than the dietary intake of BCAA, the overall dietary pattern that contributes to high BCAA plasma concentrations may modulate chronic diseases risk.


Evaluating genome architecture of a complex region via generalized bipartite matching.
With the remarkable development in inexpensive sequencing technologies and supporting computational tools, we have the promise of medicine being personalized by knowledge of the individual genome. Current technologies provide high throughput, but short reads. Reconstruction of the donor genome is based either on de novo assembly of the (short) reads, or on mapping donor reads to a standard reference. While such techniques demonstrate high success rates for inferring 'simple' genomic segments, they are confounded by segments with complex duplication patterns, including regions of direct medical relevance, like the HLA and the KIR regions.In this work, we address this problem with a method for assessing the quality of a predicted genome sequence for complex regions of the genome. This method combines two natural types of evidence: sequence similarity of the mapped reads to the predicted donor genome, and distribution of reads across the predicted genome. We define a new scoring function for read-to-genome matchings, which penalizes for sequence dissimilarities and deviations from expected read location distribution, and present an efficient algorithm for finding matchings that minimize the penalty. The algorithm is based on a formal problem, first defined in this paper, called Coverage Sensitive many-to-many min-cost bipartite Matching (CSM). This new problem variant generalizes the standard (one-to-one) weighted bipartite matching problem, and can be solved using network flows. The resulting Java-based tool, called SAGE (Scoring function for Assembled GEnomes), is freely available upon request. We demonstrate over simulated data that SAGE can be used to infer correct haplotypes of the highly repetitive KIR region on the Human chromosome 19.


Low activity of the classical complement pathway predicts short survival of patients with chronic lymphocytic leukaemia.
The activities of the classical (CP) and alternative (AP) complement pathways as well as the levels of some complement components and circulating immune complexes were measured in 43 patients with chronic lymphocytic leukaemia (CLL) between 1980 and 1984. Depressed CP activities were frequently found in these patients. Clinical course of the disease in the patients was followed until 1992, and compared with the initial complement values. During the follow-up period 36 patients died, death of 33 patients being related to the underlying disease. A strong positive correlation (P < 0.01) was found between the length of survival of the patients and the initial CP values. Patients were divided into two groups: group A, short-term survivors, i.e patients who died in CLL-related complications within 3 years after the complement measurements; and group B, long-term survivors who died > or = 4 years after the complement measurements due to any cause, or were alive at the end of the follow-up period. Average CP values in Group B were almost twice those in group A (P = 0.002), and a similar but less pronounced difference was found in C3 levels (P = 0.055). These differences were even more marked (P = 0.0006 and P = 0.0015, respectively) when only patients in Rai stage 2 and 3 were considered. Low classical pathway activities predicted short survival time: according to the logrank test, patients in Rai stage 2-3 with low (< mean - 2s.d. of the normal values), and normal CP levels survived for 2.0 +/- 1.1, and 4.6 +/- 3.0 years, respectively. All the nine and 11/13 patients with low CP and C4 levels, respectively, died within 3 years after the complement measurements were made. These findings indicate that complement measurements performed in CLL patients have a clinical value.


Delayed ultrastructural lung maturation in the fetal and newborn hypothyroid (Hyt/Hyt) mouse.
Thyroid hormones influence fetal and neonatal lung growth and maturation. However, the effect of naturally occurring, genetically determined hypo- or hyperthyroidism on fetal or neonatal lung maturation has not been examined. In the hyt/hyt mouse, primary hypothyroidism, which is characterized by a high serum TSH concentration, is transmitted as an autosomal recessive trait. It occurs due to a mutational defect in the beta-subunit of the TSH receptor. We studied the lung ultrastructure of the fetal [18-d-gestation (term = approximately 19.5 d)] and neonatal (< 1-d-old) hyt/hyt mouse. In addition, disaturated phosphatidylcholine and total phospholipid contents of newborn hyt/hyt mouse lungs were determined. Male and female hyt/hyt mice with a high serum TSH concentration were made euthyroid by adding 3,5,3'-triiodothyronine to drinking water and then mated. Balb-c mice served as euthyroid controls. Fetal and neonatal hyt/hyt mice had a higher serum TSH concentration than the Balb-c controls. Fetal hyt/hyt mouse lungs showed a large amount of intracellular glycogen and fewer lamellar bodies in epithelial type II cells compared with Balb-c fetal mouse lungs. The neonatal hyt/hyt mouse also showed signs of lung immaturity such as persistent epithelial cell glycogen, few lamellar bodies, reduced disaturated phosphatidylcholine content, and absent tubular myelin. We conclude that fetal and neonatal lung maturation is delayed in the hyt/hyt mouse with primary hypothyroidism.


The Challenge of Next Generation Sequencing in a Boy With Severe Mononucleosis and EBV-related Lymphoma.
A severe course of infectious mononucleosis should always lead up to the suspicion of a primary immunodeficiency. We describe the case of a boy with severe mononucleosis accompanied by the development of hemophagocytic lymphohistiocytosis and lymphoma. By whole exome sequencing, we identified a mutation of uncertain significance in CTPS2, a gene closely related to CTPS1, which is involved in a primary immune deficiency with susceptibility to herpesviruses. We discuss the challenge of a correct interpretation of data from whole exome sequencing, questioning whether the CTPS2 variant found in our patient is just an incidental finding or a mutation with variable penetrance.


Critical role of STAT3 in melanoma metastasis through anoikis resistance.
Anoikis is an anchorage-independent cell death. Resistance to anoikis is one of the key features of metastatic cells. Here, we analyzed the role of STAT3 in anoikis resistance in melanoma cells leading to metastasis. When grown under anchorage-independent conditions, significant proportion of cells resisted anoikis and these resistant cells had higher rate of migration and invasion as compared to the cells grown under anchorage-dependent conditions. The anoikis resistant cells also had significantly higher expression and phosphorylation of STAT3 at Y705 than the cells that were attached to the basement membrane. STAT3 inhibitors, AG 490 and piplartine (PL) induced anoikis in a concentration-dependent manner in anoikis resistant cells. Over-expression of STAT3 or treatment with IL-6 not only increased anoikis resistance, but also protected the cancer cells from PL-induced anoikis. On the other hand, silencing STAT3 decreased the potential of cancer cells to resist anoikis and to migrate. STAT3 knock-down cells and PL treated cells did not form tumors as well as failed to metastasize in SCID-NSG mice as compared to untreated anchorage-independent cells, which formed big tumors and extensively metastasized. In summary, our results for the first time establish STAT3 as a critical player that renders anoikis resistance to melanoma cells and enhance their metastatic potential.


Two polyphenol-rich Brazilian fruit extracts protect from diet-induced obesity and hepatic steatosis in mice.
Consumption of polyphenol-rich food is associated with better metabolic health. Tucum-do-Pantanal (Bactris setosa Mart) and taruma-do-cerrado (Vitex cymosa Bertero ex Spreng) are underexploited native Brazilian fruits with an important source of phytochemicals. In this study, we assessed the effects of 100 mg kg-1 tucum (TPE) and taruma (TCE) extracts on diet-induced obesity (DIO) C57BL/6J mice. After 8 weeks of daily treatment, TPE and TCE were found to significantly prevented the diet-induced body weight gain and fully protected against hepatic steatosis associated with a tendency to stimulate hepatic AMPK phosphorylation. TPE reduced visceral obesity and improved glucose metabolism as revealed by an improvement of the insulin tolerance test, a reduction in the insulin fasting level, and a decreased glucose-induced hyperinsulinemia during an oral glucose tolerance test. TPE and TCE showed promising effects on the treatment of obesity and NAFLD, furthermore, TPE on insulin resistance.


Kif14 overexpression accelerates murine retinoblastoma development.
The mitotic kinesin KIF14 has an essential role in the recruitment of proteins required for the final stages of cytokinesis. Genomic gain and/or overexpression of KIF14 has been documented in retinoblastoma and a number of other cancers, such as breast, lung and ovarian carcinomas, strongly suggesting its role as an oncogene. Despite evidence of oncogenic properties in vitro and in xenografts, Kif14's role in tumor progression has not previously been studied in a transgenic cancer model. Using a novel Kif14 overexpressing, simian virus 40 large T-antigen retinoblastoma (TAg-RB) double transgenic mouse model, we aimed to determine Kif14's role in promoting retinal tumor formation. Tumor initiation and development in double transgenics and control TAg-RB littermates were documented in vivo over a time course by optical coherence tomography, with subsequent ex vivo quantification of tumor burden. Kif14 overexpression led to an accelerated initiation of tumor formation in the TAg-RB model and a significantly decreased tumor doubling time (1.8 vs. 2.9 weeks). Moreover, overall percentage tumor burden was also increased by Kif14 overexpression. These data provide the first evidence that Kif14 can promote tumor formation in susceptible cells in vivo.


Decoding onset and direction of movements using Electrocorticographic (ECoG) signals in humans.
Communication of intent usually requires motor function. This requirement can be limiting when a person is engaged in a task, or prohibitive for some people suffering from neuromuscular disorders. Determining a person's intent, e.g., where and when to move, from brain signals rather than from muscles would have important applications in clinical or other domains. For example, detection of the onset and direction of intended movements may provide the basis for restoration of simple grasping function in people with chronic stroke, or could be used to optimize a user's interaction with the surrounding environment. Detecting the onset and direction of actual movements are a first step in this direction. In this study, we demonstrate that we can detect the onset of intended movements and their direction using electrocorticographic (ECoG) signals recorded from the surface of the cortex in humans. We also demonstrate in a simulation that the information encoded in ECoG about these movements may improve performance in a targeting task. In summary, the results in this paper suggest that detection of intended movement is possible, and may serve useful functions.


Discovery of Potential Inhibitors for RNA-Dependent RNA Polymerase of Norovirus: Virtual Screening, and Molecular Dynamics.
Noroviruses are non-enveloped viruses with a positive-sense single-stranded RNA (ssRNA) genome belonging to the genus Norovirus, from the family Caliciviridae, which are accountable for acute gastroenteritis in humans. The Norovirus genus is subdivided into seven genogroups, i.e., (GI-GVII); among these, the genogroup II and genotype 4 (GII.4) strains caused global outbreaks of human norovirus (HuNov) disease. The viral genome comprises three open reading frames (ORFs). ORF1 encodes the nonstructural polyprotein that is cleaved into six nonstructural proteins, which include 3C-like cysteine protease (3CLpro) and a viral RNA-dependent RNA polymerase. ORF2 and ORF3 encode the proteins VP1 and VP2. The RNA-dependent RNA polymerase (RdRp) from noroviruses is one of the multipurpose enzymes of RNA viruses vital for replicating and transcribing the viral genome, making the virally encoded enzyme one of the critical targets for the development of novel anti-norovirus agents. In the quest for a new antiviral agent that could combat HuNov, high throughput virtual screening (HTVS), combined with e-pharmacophore screening, was applied to screen compounds from the PubChem database. CMX521 molecule was selected as a prototype for a similarity search in the PubChem online database. Molecular dynamics simulations were employed to identify different compounds that may inhibit HuNov. The results predicted that compound CID-57930781 and CID-44396095 formed stable complexes with MNV-RdRp within 50 ns; hence, they may signify as promising human norovirus inhibitors.


Diazepam enhances etoposide-induced cytotoxicity in U-87 MG human glioma cell line.
Various approaches might be employed in an effort to increase efficacy of the chemotherapeutic treatment of cancer. Recently, various modulators of anticancer therapy effectiveness have been studied. Antiproliferative effects of peripheral benzodiazepine receptor (PBR) ligands might be exploited to enhance cytotoxic effect of a chemotherapeutic drug towards cancer cells. In this work, we sought to enhance cytotoxic effect of etoposide (VP-16) by a PBR ligand, diazepam (DZ) in U-87 MG human glioma cells. Cytotoxicity of VP-16, DZ and their combinations was assessed by using the microculture MTT assay. Cell survival, effective concentrations (EC) and the onset of cytotoxic effect were determined. After 72 h of cultivation, survival of U-87 MG cells was reduced to 57 +/- 7% in the presence of VP-16 at 12.5 microg/mL alone, whereas DZ at 10-4 mol/L alone caused 28 +/- 6% reduction in cell survival. Coincubation of VP-16 at 12.5 microg/mL with DZ at 10-4 mol/L led to a further decrease in cell survival to 45 +/- 6%. Furthermore, DZ at 10-4 mol/L significantly decreased effective concentrations, EC10, EC30 and EC50, of VP-16 and the dose-response curves were shifted to the left. Addition of DZ at 10-4 mol/L to VP-16 also facilitated the onset of its cytotoxic effect. The same decrease in survival was thus achieved approximately 30 h earlier in comparison with VP-16 alone. However, DZ at 10-9 mol/L failed both to exert any effect on glioma cells survival and enhance cytotoxic effect of VP-16. DZ at 10-4 mol/L was capable of both reducing U-87 MG glioma cells survival when applied alone and also enhancing the cytotoxic effect of VP-16. No such observation was made for the lower concentrations of DZ. Potential implementation of diazepam in the antiglioma/anticancer armamentarium awaits further experimentation but phase I and phase II clinical trials could be suggested.


Peri-aortic fibrosis: progression and regression.
The natural history and aetiology of peri-aortic fibrosis is unknown. In one of our patients in whom steroid treatment had successfully controlled the progression of the disease' cessation of treatment was associated with recurrence of the fibrosis, progressive ureteric obstruction and dilatation of the aorta. In another patient, graft replacement of the aorta was associated with regression of the fibrosis but in a third patient replacement of the aorta did not prevent progression to complete ureteric obstruction. It would seem that it is the presence of the fibrosis that engenders the aortic dilatation not the presence of a patent aorta, dilated or not.


Leveling the Playing Field: A Comparative Reevaluation of Unmodified Eye Tracking as an Input and Interaction Modality for VR.
In this study, we establish a much-needed baseline for evaluating eye tracking interactions using an eye tracking enabled Meta Quest 2 VR headset with 30 participants. Each participant went through 1098 targets using multiple conditions representative of AR/VR targeting and selecting tasks, including both traditional standards and those more aligned with AR/VR interactions today. We use circular white world-locked targets, and an eye tracking system with sub-1-degree mean accuracy errors running at approximately 90Hz. In a targeting and button press selection task, we, by design, compare completely unadjusted, cursor-less, eye tracking with controller and head tracking, which both had cursors. Across all inputs, we presented targets in a configuration similar to the ISO 9241-9 reciprocal selection task and another format with targets more evenly distributed near the center. Targets were laid out either flat on a plane or tangent to a sphere and rotated toward the user. Even though we intended this to be a baseline study, we see unmodified eye tracking, without any form of a cursor, or feedback, outperformed the head by 27.9% and performed comparably to the controller (5.63% decrease) in throughput. Eye tracking had improved subjective ratings relative to head in Ease of Use, Adoption, and Fatigue (66.4%, 89.8%, and 116.1% improvements, respectively) and had similar ratings relative to the controller (reduction by 4.2%, 8.9%, and 5.2% respectively). Eye tracking had a higher miss percentage than controller and head (17.3% vs 4.7% vs 7.2% respectively). Collectively, the results of this baseline study serve as a strong indicator that eye tracking, with even minor sensible interaction design modifications, has tremendous potential in reshaping interactions in next-generation AR/VR head mounted displays.


Integrative Analysis of Proteome and Ubiquitylome Reveals Unique Features of Lysosomal and Endocytic Pathways in Gefitinib-Resistant Non-Small Cell Lung Cancer Cells.
Non-small cell lung cancer (NSCLC) patients carrying EGFR activating mutations treated with gefitinib, a tyrosine kinase inhibitor, will develop drug resistance. Ubiquitylation is one of major posttranslational modifications of proteins affecting the stability or function of proteins. However, the role of protein ubiquitylation in gefitinib resistance is poorly understood. To systematically identify the global change in protein expression and ubiquitylation during gefitinib resistance, a quantitative global proteome and ubiquitylome study in a pair of gefitinib-resistant and sensitive NSCLC cells is carried out. Altogether, changes in expression of 3773 proteins are quantified, and changes in ubiquitylation of 2893 lysine sites in 1415 proteins are measured in both cells. Interestingly, lysosomal and endocytic pathways, which are involved in autophagy regulation, are enriched with upregulated proteins or ubiquitylated proteins in gefitinib-resistant cells. In addition, HMGA2 overexpression or ALOX5 knockdown suppresses gefitinib resistance in NSCLC cells by inhibiting autophagy. Overall, these results reveal the previously unknown global ubiquitylome and proteomic features associated with gefitinib resistance, uncover the opposing roles of HMGA2 or ALOX5 in regulating gefitinib resistance and autophagy, and will help to identify new therapeutic targets in overcoming gefitinib resistance.


Bond strength test: role of operator skill.
Test method, substrate material and operator variability are factors known to affect bond strength test outcomes. The aim of this study was to determine if, with increasing experience, individual operator skill in material handling influenced microshear bond strength outcomes. This retrospective study used data collected from six preliminary tests carried out successively on enamel microshear bond strength for two adhesives: Clearfil SE Bond (Kuraray) and Adper Single Bond 2 (3M ESPE) by one operator. Mean and median microshear bond strength values, standard deviations and coefficients of variation for each adhesive test group were calculated and assessed. A gradual increase in mean microshear bond strengths, a decrease in standard deviations and coefficients of variation over time for both adhesives and a progressively more uniform distribution of microshear bond strength results recorded in individual test sets was observed. The results suggest an improvement in bonding reliability as experience is gained. Operator skill in material handling appears to play a critical role in determining the outcome of bond strength testing. This same skill is probably relevant when using an adhesive for the first time clinically. It is important that clinicians become familiar with new adhesives in order to achieve optimal outcomes.


Bioactive sesquiterpene aryl esters from the culture broth of Armillaria sp.
Two new compounds, 10-dehydroxymelleolide D (1) and 13-hydroxymelleolide K (2), along with seven known compounds, 5'-O-methylmelledonal (3), melleolide D (4), 13-hydroxydihydromelleolide (5), melleolide (6), armillarinin (7), armillaridin (8), and armillarikin (9), were isolated from the culture broth of Armillaria sp. Their structures were determined by spectroscopic data analysis. All the compounds inhibited plant growth of lettuce. Melleolide (6) and armillarikin (9) inhibited mycelial growth of Coprinopsis cinerea and/or Flammulina velutipes.


Transcriptome-wide investigation of stop codon readthrough in Saccharomyces cerevisiae.
Translation of mRNA into a polypeptide is terminated when the release factor eRF1 recognizes a UAA, UAG, or UGA stop codon in the ribosomal A site and stimulates nascent peptide release. However, stop codon readthrough can occur when a near-cognate tRNA outcompetes eRF1 in decoding the stop codon, resulting in the continuation of the elongation phase of protein synthesis. At the end of a conventional mRNA coding region, readthrough allows translation into the mRNA 3'-UTR. Previous studies with reporter systems have shown that the efficiency of termination or readthrough is modulated by cis-acting elements other than stop codon identity, including two nucleotides 5' of the stop codon, six nucleotides 3' of the stop codon in the ribosomal mRNA channel, and stem-loop structures in the mRNA 3'-UTR. It is unknown whether these elements are important at a genome-wide level and whether other mRNA features proximal to the stop codon significantly affect termination and readthrough efficiencies in vivo. Accordingly, we carried out ribosome profiling analyses of yeast cells expressing wild-type or temperature-sensitive eRF1 and developed bioinformatics strategies to calculate readthrough efficiency, and to identify mRNA and peptide features which influence that efficiency. We found that the stop codon (nt +1 to +3), the nucleotide after it (nt +4), the codon in the P site (nt -3 to -1), and 3'-UTR length are the most influential features in the control of readthrough efficiency, while nts +5 to +9 had milder effects. Additionally, we found low readthrough genes to have shorter 3'-UTRs compared to high readthrough genes in cells with thermally inactivated eRF1, while this trend was reversed in wild-type cells. Together, our results demonstrated the general roles of known regulatory elements in genome-wide regulation and identified several new mRNA or peptide features affecting the efficiency of translation termination and readthrough.


Point-of-sale nutrition information interventions in food retail stores to promote healthier food purchase and intake: A systematic review.
Providing simple information that identifies healthier/less healthy products at the point-of-sale has been increasingly recognized as a potential strategy for improving population diet. This review evaluated the effect on healthiness of food purchasing/intake of interventions that identify specific products as healthier/less healthy at the point-of-sale in food retail settings. Five databases were searched for peer-reviewed randomized controlled or quasi-experimental trials published 2000-2020. Effects on primary outcomes of the 26 eligible studies (322 stores and 19,002 participants) were positive (n = 14), promising (effective under certain conditions; n = 3), mixed (different effect across treatment arms/outcomes; n = 4), null (n = 3), negative (n = 1), or unclear (n = 1). Shelf-label studies (three studies of two rating systems across all products) were positive. Technology-delivered (mobile applications/podcast/kiosk) interventions were positive (n = 3/5) or promising/mixed (n = 2/5). In-store displays (n = 16) had mixed effectiveness. Interventions provided information on targeted healthier products only (n = 17), unhealthy products only (n = 1), both healthy and unhealthy (n = 2), and across all products (n = 5). No patterns were found between behavior change technique used and effectiveness. Study quality was mixed. These findings indicate that point-of-sale interventions identifying healthy/unhealthy options can lead to healthier customer purchasing behavior, particularly those delivered using shelf-labels or technology. Further research on discouraging unhealthy foods is needed.


Digital Health during COVID-19: Informatics Dialogue with the World Health Organization.
On December 16, 2020 representatives of the International Medical Informatics Association (IMIA), a Non-Governmental Organization in official relations with the World Health Organization (WHO), along with its International Academy for Health Sciences Informatics (IAHSI), held an open dialogue with WHO Director General (WHO DG) Tedros Adhanom Ghebreyesus about the opportunities and challenges of digital health during the COVID-19 global pandemic. The aim of this paper is to report the outcomes of the dialogue and discussions with more than 200 participants representing different civil society organizations (CSOs). The dialogue was held in form of a webinar. After an initial address of the WHO DG, short presentations by the panelists, and live discussions between panelists, the WHO DG and WHO representatives took place. The audience was able to post questions in written. These written discussions were saved with participants' consent and summarized in this paper. The main themes that were brought up by the audience for discussion were: (a) opportunities and challenges in general; (b) ethics and artificial intelligence; (c) digital divide; (d) education. Proposed actions included the development of a roadmap based on the lessons learned from the COVID-19 pandemic. Decision making by policy makers needs to be evidence-based and health informatics research should be used to support decisions surrounding digital health, and we further propose next steps in the collaboration between IMIA and WHO such as future engagement in the World Health Assembly.


The effects of appreciative inquiry interviews on staff in the U.K. National Health Service.
To obtain preliminary data on the short- and medium-term effects and personal acceptability of appreciative inquiry (AI) in staff development in health care. AI is a non-problem-solving management approach focusing on developing current successes into the future through reflection at individual and group level. Individual one-hour interviews were undertaken with nursing staff on a national paediatric liver in-patient ward. They were asked to recount stories based on their experiences of successful delivery of health care, with active listening, followed by reflection on the process. A total of 32 staff members took part with only two refusals. Data were written and analysed by an open coding method. Follow up was obtained two years later using a written, open question method. The process was emotional-but well received. Staff described quality in interpersonal interactions, preventing errors and engaging their personal values in their work. No improvement in recruitment or retention was shown but a high level of sickness absence fell significantly during the period of the project. Two years later, significant positive effects were recalled and attributed to the interviews by many respondents. AI appears a cost-effective way of connecting professionals' motivation toward quality in their work with strategic intentions. The interviewer was a medical consultant and ward manager, implying either that the interviews could have worked as a form of managerial supervision or improvements could be a Hawthorne effect. Other unknown influences were likely to be occurring on the ward during the study period. Short AI interventions on an individual basis can change sickness absence, at least while the interventions are continuing. It is an important tool for staff motivation with the potential for connecting strategic with micro-operational levels. AI is an approach to NHS management with wide application including appraisal, personal development and mentoring. It can be a positive introduction to reflective practice. AI is gaining recognition for its value in staff and service development in health care. The paper shows service and personal effects, cost-effectiveness and illustrates how to use AI for these purposes.


High-resolution 23Na-NMR studies of human erythrocytes: use of aqueous shift reagents.
Aqueous shift reagents were used to clearly distinguish intra-and extracellular 23Na-nuclear magnetic resonance (NMR) signals in samples consisting of whole blood or suspensions of washed human erythrocytes (both fresh and outdated). The lanthanide chelates Dy(PPP)2(7-) and Tm( TTHA )3- were used to shift the extracellular signals upfield, and Dy( TTHA )3- and Tm(PPP)2(7-) were similarly used to shift extracellular resonances downfield. The absolute intensities of the signals were used along with the measured hematocrit to simultaneously determine the intra- and extracellular Na+ concentrations. The results were generally within 5% of the values determined by more time-consuming centrifugation-flame emission photometry measurements on the same samples. Thus the 23Na-NMR signals from both intra- and extracellular cations suffer no NMR invisibility within experimental error. The lower level of intracellular Na+ in fresh erythrocytes (less than 12 mM) is easily distinguished from the higher level (approximately 30 mM) in erythrocytes that have been stored (in the cold) outside the body for some weeks.