Long chain polyunsaturated fatty acids alter membrane-bound RANK-L expression and osteoprotegerin secretion by MC3T3-E1 osteoblast-like cells.
Inflammation triggers an increase in osteoclast (bone resorbing cell) number and activity. Osteoclastogenesis is largely controlled by a triad of proteins consisting of a receptor (RANK), a ligand (RANK-L) and a decoy receptor (osteoprotegerin, OPG). Whilst RANK is expressed by osteoclasts, RANK-L and OPG are expressed by osteoblasts. The long chain polyunsaturated fatty acid (LCPUFA) arachidonic acid (AA, 20:4n-6) and its metabolite prostaglandin E2 (PGE2), are pro-inflammatory and PGE2 is a potent stimulator of RANKL expression. Various LCPUFAs such as eicosapentaenoic acid (EPA, 20:5n-3), docosahexaenoic acid (DHA, 22:6n-3) and gamma-linolenic acid (GLA, 18:3n-6) have anti-inflammatory activity. We aimed to determine if AA itself can stimulate RANKL expression and whether EPA, DHA and GLA inhibit RANKL expression in osteoblasts. MC3T3-E1/4 osteoblast-like cells were cultured under standard conditions with each of the LCPUFAs (5microg/ml) for 48h. Membrane-bound RANKL expression was measured by flow cytometry and OPG secretion measured by ELISA. In a second experiment, RANKL expression in MC3T3-E1/4 cells was stimulated by PGE2 treatment and the effect of EPA, DHA and GLA on membrane-bound RANKL expression and OPG secretion determined. The percentage of RANKL-positive cells was higher (p<0.05) than controls following treatment with AA or GLA but not after co-treatment with the cyclooxygenase inhibitor, indomethacin. DHA and EPA had no effect on membrane-bound RANKL expression under standard cell culture conditions. Secretion of OPG was lower (p<0.05) in AA-treated cells but not significantly different from controls in GLA, EPA or DHA treated cells. Treatment with prostaglandin E2 (PGE2) resulted in an increase (p<0.05) in the percentage of RANK-L positive cells and a decrease (p<0.05) in mean OPG secretion. The percentage of RANKL positive cells was significantly lower following co-treatment with PGE2 and either DHA or EPA compared to treatment with PGE2 alone. Mean OPG secretion remained lower than controls in cells treated with PGE2 regardless of co-treatment with EPA or DHA. Results from this study suggest COX products of GLA and AA induce membrane-bound RANKL expression in MC3T3-E1/4 cells. EPA and DHA have no effect on membrane-bound RANKL expression in cells cultured under standard conditions however both EPA and DHA inhibit the PGE2-induced increase in RANKL expression in MC3T3-E1/4 cells.


Survival and Immunosuppression Induced by Hepatocyte Growth Factor in Chronic Lymphocytic Leukemia.
Chronic lymphocytic leukemia (CLL), the most common leukemia among adults in the western world, is characterized by a progressive accumulation of relatively mature CD5+ B cells in peripheral blood, lymph nodes and bone marrow. Despite much recent advancement in therapy, CLL is still incurable. Lymph nodes and bone marrow represent sanctuary sites preserving leukemic cells from spontaneous or drug-induced apoptosis, and infiltration of leukemic cells in these districts correlates with clinical stages and prognosis. The central role played by the microenvironment in the disease has become increasingly clear. Different chemokines (CXCL12, CXCL13, CCL19, CCL21) may in fact participate in attracting CLL cells into bone marrow and lymph nodes, where various factors, such as IL-15 and CXCL12, enhance leukemic cells survival. Recently, we have suggested that hepatocyte growth factor (HGF), produced by microenvironmental stromal cells, can contribute to CLL pathogenesis. We have demonstrated that HGF exerts a double effect on CLL B cells through the interaction with its receptor c- MET; HGF, infact, protects CLL B cells, which are c-MET+, from apoptosis, and also polarizes mono/macrophages towards the M2 phenotype, thus facilitating the evasion of the CLL clone from immune control. This double effect appears mediated by the activation of two major signaling pathways: STAT3TYR705 and AKT. The aim of this review is to summarize data on HGF and c-MET expression in normal B cells and in B cell malignancies, with a particular emphasis on our results obtained in CLL. Altogether, the observations described here suggest that the HGF/c-MET axis may have a prominent role in malignancy progression further indicating novel potential therapeutic options aimed to block HGF-induced signaling pathways in B lymphoproliferative disorders.


Autism, Gastrointestinal Symptoms and Modulation of Gut Microbiota by Nutritional Interventions.
Autism spectrum disorder (ASD) is a complex behavioral syndrome that is characterized by speech and language disorders, intellectual impairment, learning and motor dysfunctions. Several genetic and environmental factors are suspected to affect the ASD phenotype including air pollution, exposure to pesticides, maternal infections, inflammatory conditions, dietary factors or consumption of antibiotics during pregnancy. Many children with ASD shows abnormalities in gastrointestinal (GI) physiology, including increased intestinal permeability, overall microbiota alterations, and gut infection. Moreover, they are "picky eaters" and the existence of specific sensory patterns in ASD patients could represent one of the main aspects in hampering feeding. GI disorders are associated with an altered composition of the gut microbiota. Gut microbiome is able to communicate with brain activities through microbiota-derived signaling molecules, immune mediators, gut hormones as well as vagal and spinal afferent neurons. Since the diet induces changes in the intestinal microbiota and in the production of molecules, such as the SCFA, we wanted to investigate the role that nutritional intervention can have on GI microbiota composition and thus on its influence on behavior, GI symptoms and microbiota composition and report which are the beneficial effect on ASD conditions.


Evaluation of a German version of the Strengthening Families Programme 10-14: a randomised controlled trial.
The purpose of this study is to evaluate the effects of a German adaptation of the Strengthening Families Programme 10-14 (SFP 10-14; Familien Stärken). A multi-centre randomised controlled trial comparing the German SFP version consisting of seven sessions and four booster-sessions with a minimal intervention on parenting as control condition. Outcomes comprise measures of adolescent substance use (initiation) and behaviour problems and are assessed at baseline, after programme delivery and at 6- and 18-month follow-ups. Primary outcomes were lifetime tobacco, alcohol and cannabis use at 18 months. Data of n = 292 families were analysed using baseline adjusted logistic regressions and mixed models. We observed reduced rates of lifetime tobacco use in analyses with follow-up respondents, but not in data using the complete intention to treat sample with multiple imputation estimates for missing data. Parents reported fewer adolescent behaviour problems in analyses with the total sample and multiple imputed data, but not in data with follow-up respondents only. There were no other significant effects of SFP 10-14. Overall the medium size effects found in previous US trials could not be replicated in a German context.


Obstructive Sleep Apnea in Posttraumatic Stress Disorder Comorbid With Mood Disorder: Significantly Higher Incidence Than in Either Diagnosis Alone.
To examine the correlations between obstructive sleep apnea (OSA) and psychiatric disorders such as major depressive disorder (MDD), posttraumatic stress disorder (PTSD), or bipolar disorder (BD) and whether comorbid psychiatric diagnosis increases the risk of OSA. This retrospective chart review study included all patients (N = 413) seen within a randomly selected 4-month period (August 2014 to November 2014) in a Veterans Administration outpatient psychiatry clinic. Patients were screened for symptoms of OSA with the STOP-BANG Questionnaire. Those with a positive screen were referred to the sleep clinic for confirmation of the diagnosis by polysomnogram (PSG). Frequency of PSG-confirmed OSA was correlated with different psychiatric disorders and comorbid psychiatric diagnoses. The study showed a high prevalence of OSA in psychiatric patients, particularly with MDD (37.8%) and PTSD (35.5%) and less so with BD (16.7%). Among all patients with OSA (n = 155), those with comorbid BD and PTSD had a significantly higher rate of OSA than those with BD alone (χ² = 7.28, P < .05) but not with PTSD alone. We also found a statistically significant higher incidence of OSA in male veterans with either MDD comorbid with PTSD (χ² = 3.869, P < .05) or BD comorbid with PTSD (χ² = 6.631, P < .05) compared with either mood disorder or PTSD alone. The study showed a high prevalence of OSA in psychiatric patients, particularly in those with PTSD and MDD and less so with BD. There was a statistically significant increase in the incidence of OSA in male veterans with either BD with comorbid PTSD or MDD with comorbid PTSD..


One-pot stirring-free synthesis of silver nanowires with tunable lengths and diameters via a Fe3+ & Cl- co-mediated polyol method and their application as transparent conductive films.
The properties of nanomaterials are highly dependent on their size, shape and composition. Compared with zero-dimensional nanoparticles, the increased dimension of a one-dimensional silver nanowire (AgNW/Ag NW) leads to extra challenges on synthesizing it with controllable sizes. Here, a convenient way for the synthesis of AgNWs with tunable sizes has been developed simply by adjusting the amount of salt additives, i.e., ferric chloride (FeCl3), or Fe(NO3)3 & KCl. The average diameter and length of nanowires are readily tailored within 45-220 nm and 10-230 μm, respectively. The distinctive roles of Fe3+ and Cl- played during the growth stages of Ag NWs were revealed by comparative experiments and a heterogeneous nucleation model with the assistance of oxidative etching was proposed to elucidate the growth mechanism. Afterwards, transformations in XRD patterns from nanometer-size effects and quantitative relation for size-dependent peak wavelength of surface plasmon resonances (SPRs) in UV-vis spectroscopy of these nanowires were studied. In addition, as transparent conductive materials (TCMs), these metal nanowires were utilized to fabricate transparent conductive films (TCFs), and the effects of their diameters and lengths were elucidated. Very/ultra-long nanowires with a high aspect ratio up to 1600 achieved impressive properties of R = 12.4 ohm sq-1 at T% = 90.1% without any post treatment. This facile method for the size-tunable growth of uniform AgNWs with high yield is attractive and ready to be home-made, which is believed to promote research in their potential applications, especially in optoelectronic devices and flexible electronics.


A physical model of microtubule sliding in ciliary axonemes.
Ciliary movement is caused by coordinated sliding interactions between the peripheral doublet microtubules of the axoneme. In demembranated organelles treated with trypsin and ATP, this sliding can be visualized during progressive disintegration. In this paper, microtubule sliding behavior resulting from various patterns of dynein arm activity and elastic link breakage is determined using a simplified model of the axoneme. The model consists of a cylindrical array of microtubules joined, initially, by elastic links, with the possibility of dynein arm interaction between microtubules. If no elastic links are broken, sliding can produce stable distortion of the model, which finds application to straight sections of a motile cilium. If some elastic links break, the model predicts a variety of sliding patterns, some of which match, qualitatively, the observed disintegration behavior of real axonemes. Splitting of the axoneme is most likely to occur between two doublets N and N + 1 when either the arms on doublet N + 1 are active and arms on doublet N are inactive or arms on doublet N - 1 are active while arms on doublet N are inactive. The analysis suggests further experimental studies which, in conjunction with the model, will lead to a more detailed understanding of the sliding mechanism, and will allow the mechanical properties of some axonemal components to be evaluated.


Effect of common comparators in indirect comparison analysis of the effectiveness of different inhaled corticosteroids in the treatment of asthma.
Indirect comparison (IC) and direct comparison (DC) of four inhaled corticosteroid (CS) treatments for asthma were conducted, and the factors that may influence the results of IC were investigated. Among those factors, we focused on the effect of common comparator selection in the treatment of asthma, where little control group bias or placebo effect is expected. IC and DC were conducted using the change from baseline in forced expiratory volume in 1 s (FEV1(L)) as an outcome parameter. Differences between inhaled CS were evaluated to compare the results of IC and DC. As a common comparator for IC, placebo (PLB) or mometasone (MOM) was selected. Whether the results of IC are affected by the selection of a common comparator and whether the results of IC and DC are consistent were examined. 23 articles were identified by a literature search. Our results showed that ICs yielded results similar to DCs in the change from baseline of FEV1(L). No statistically significant difference was observed in inconsistency analysis between ICs and DCs. It was clinically and statistically confirmed that ICs with PLB and those with MOM did not differ in terms of the results of FEV1(L) analysis in this dataset. This study demonstrated that ICs among inhaled CS can deliver results consistent with those of DCs when using the change from baseline in FEV1(L) as an outcome parameter in asthma patients. It was also shown that using an active comparator has similar results if there is no effect of control group bias. It should be emphasized that the investigation of control group bias is a key factor in conducting relevant ICs so that an appropriate common comparator can be selected.


Methylation of a phosphatase specifies dephosphorylation and degradation of activated brassinosteroid receptors.
Internalization of cell surface receptors, followed by either recycling back to the plasma membrane or degradation, is crucial for receptor homeostasis and signaling. The plant brassinosteroid (BR) receptor, BRASSINOSTEROID INSENSITIVE 1 (BRI1), undergoes constitutive cycling between the plasma membrane and the internal membranes. We show that protein phosphatase 2A (PP2A) dephosphorylated BRI1 and that Arabidopsis thaliana rcn1, a mutant for a PP2A subunit, caused an increase in BRI1 abundance and BR signaling. We report the identification, in A. thaliana, of a suppressor of bri1, sbi1, which caused selective accumulation of BR-activated BRI1, but not the BR co-receptor BAK1 (BRI1-ASSOCIATED KINASE 1), in the membranous compartment. SBI1 mRNA was induced by BRs, and SBI1 encodes a leucine carboxylmethyltransferase (LCMT) that methylated PP2A and controlled its membrane-associated subcellular localization. We propose that BRs increase production of SBI1, which methylates PP2A, thus facilitating its association with activated BRI1. This leads to receptor dephosphorylation and degradation, and thus to the termination of BR signaling.


Understanding variability in the benefits of N2-fixation in soybean-maize rotations on smallholder farmers' fields in Malawi.
Soybean production can contribute to the nitrogen economy of smallholder farming systems, but our understanding of factors explaining variability in nitrogen fixation and rotational benefits across farms and regions is limited. Biological nitrogen fixation (BNF) was quantified with the natural abundance method in 150 farmer-managed soybean plots under different varieties and inputs in Dowa, Mchinji and Salima districts of Malawi. Soybean yielded on average 1.2 t ha-1 grain and the above-ground biomass at mid pod filling (R5.5) was 2.8 t ha-1 and contained in total 63 kg ha-1 nitrogen derived from the atmosphere (Ndfa). Locally sourced varieties obtained a larger %Ndfa (65%) than the 'improved' variety Nasoko (53%). The %Ndfa was positively associated with soil sand content, sowing date, plant population and biomass accumulation, but it was not affected by inoculation with rhizobia or the combination of inoculation and NPK fertiliser application. Quantities of N2 fixed differed between regions and years, and was enhanced by applying inoculant and fertiliser together, leading to more biomass accumulation and larger grain yields. Soil available P and exchangeable K contents also increased the total amount of N2 fixed. In a related trial, continuous maize yields were compared with maize following soybean in 53 farmer-managed fields. Average yield in continuous maize was 2.5 t ha-1, while maize after soybean produced 3.5 t ha-1 (139% of continuous maize). Farmers with higher maize yields, who applied external nutrient inputs, and with a larger value of household assets achieved greater yield responses to rotation with soybean. A relative yield increase of more than 10% was observed on 59, 90 and 77% of the fields in Dowa, Mchinji and Salima respectively. We conclude that fields of soybean and maize that receive adequate nutrient inputs and good management to ensure good yields benefit most in terms of quantities of N2 fixed by the legume and the yield response of the following maize crop. The results suggest that the promotion of soybean-maize rotations should be done through an integrated approach including the promotion of appropriate soil and crop management techniques. Furthermore, they suggest that wealthier households are more likely to apply adequate nutrient inputs and good crop management practices and are likely to receive larger maize yield responses to the incorporation of soybean.


Management of neonatal abstinence syndrome: a national survey and review of practice.
To ascertain the present management of neonatal abstinence syndrome (NAS) in neonatal units in the United Kingdom (UK) and Ireland. Postal questionnaire to 235 neonatal units, with telephone follow-up of non-respondents. The response rate was 90%, and 96% of respondents had a formal NAS guideline. The median number of infants treated annually for NAS was 6 (range 1-100). The method of Finnegan was the most widely used scoring system (52%). Morphine sulphate was the most commonly used first line agent for both opiate (92%) and polysubstance (69%) withdrawal. Dosing regimens varied widely. Units using a maximum daily morphine dose of <400 microg/kg/day were more likely to require the addition of a second agent (76% vs 58%, p = 0.027). Phenobarbitone was the drug of choice to treat seizures secondary to both opiate and polydrug withdrawal in 73% and 81% of units, respectively. 29% of units allowed infants to be discharged home on medication. 58% of these allowed administration of opiates in the community and in almost half of cases this was managed by a parent. Mothers on methadone whose serology was positive for hepatitis B and/or C were four times more likely to be discouraged from breastfeeding. The majority of units currently use an opiate as the drug of first choice as recommended. Doses utilised and second agents added vary significantly between units. Many of our findings reflect the lack of high-quality randomised studies regarding management of NAS.


Human stromelysin gene promoter activity is modulated by transcription factor ZBP-89.
Matrix metalloproteinase expression is under strict regulation in physiological conditions. Disruption of the regulatory mechanisms can lead to tissue destruction and is associated with tumour invasion and metastasis. Using the one-hybrid assay technique with a cis-element in the promoter region of the stromelysin (matrix metalloproteinase-3) gene, a cDNA encoding a transcription factor termed ZBP-89 was obtained. The interaction between ZBP-89 and the stromelysin promoter element was confirmed by electrophoretic mobility shift assays with a recombinant ZBP-89. Reporter gene expression under the control of the stromelysin promoter in transiently transfected cells was significantly increased when the cells were cotransfected with a ZBP-89 expression construct. These results indicate that ZBP-89 interacts with the stromelysin promoter and upregulates its activity. As ZBP-89 expression is known to be increased in gastric carcinoma cells, induction of stromelysin expression may be a significant factor in tumour metastasis.


Homelessness-Related Traumatic Events and PTSD Among Women Experiencing Episodes of Homelessness in Three U.S. Cities.
In this article, we report the prevalence of traumatic events (TEs), lifetime and 12-month posttraumatic stress disorder (PTSD) among 148 women experiencing homelessness in 3 midsized cities in the United States (Omaha, NE, Pittsburgh, PA, and Portland, OR). The women ranged in age from 19 to 54 years with an average age of 38.89 years (SD = 10.18). The sample was 42.6% White/European American. We investigated the mediation of distal TEs (i.e., childhood maltreatment) by more proximal TEs associated with being homeless (i.e., homelessness-related stressors) for meeting diagnostic criteria for 12-month PTSD. Results indicated that 42.6% of the women met criteria for lifetime PTSD and 39.7% met criteria for past-year PTSD. The number of TEs reported ranged from 0 to 16 in order of prevalence with a median of 6 TEs. The correlations between childhood maltreatment and 12-month PTSD ranged from .16 to .20 and the correlations between homelessness-related stressors and 12-month PTSD ranged from .21 to .30. The mediation analysis was consistent with the association between childhood maltreatment and past-year PTSD being fully mediated by homelessness-related trauma.


New therapeutic strategies in radiation therapy.
Radiation therapy has been an integral part of curative cancer therapy for many decades. The tolerance of normal tissues traversed by radiation and resistant tumor cell populations traditionally have limited the radiocurability to certain tumors, especially with higher clinical stages. However, research in radiobiology and radiation physics is providing ways to increase cure while limiting morbidity. Computer-controlled dynamic treatment, radioprotector drugs, hyperbaric oxygen, carbogen breathing during irradiation, particle irradiation, and hypoxic cell sensitizing drugs are presently undergoing clinical evaluation with some encouraging preliminary results. We review the fundamental concepts underlying these clinical trials and analyze the results.


Promoting social-emotional competence: An evaluation of the elementary version of Second Step®.
Research has consistently linked social-emotional skills to important educational and life outcomes. Many children begin their school careers, however, without the requisite social and emotional skills that facilitate learning, which has prompted schools nationwide to adopt specific curricula to teach students the social-emotional skills that enable them to maintain optimal engagement in the learning process. Second Step® is one of the most widely disseminated social-emotional learning (SEL) programs; however, its newly revised version has never been empirically evaluated. The purpose of this study was to conduct a randomized controlled trial investigating the impact of the 4th Edition Second Step® on social-behavioral outcomes over a 1-year period when combined with a brief training on proactive classroom management. Participants were kindergarten to 2nd grade students in 61 schools (321 teachers, 7300 students) across six school districts. Hierarchical models (time×condition) suggest that the program had few main effects from teacher-reported social and behavioral indices, with small effect sizes. The majority of significant findings were moderated effects, with 8 out of 11 outcome variables indicating the intervention-produced significant improvements in social-emotional competence and behavior for children who started the school year with skill deficits relative to their peers. All the significant findings were based on teacher-report data highlighting a need for replication using other informants and sources of data. Findings provide program validation and have implications for understanding the reach of SEL programs.


Efficiency of exonucleolytic action of apurinic/apyrimidinic endonuclease 1 towards matched and mismatched dNMP at the 3' terminus of different oligomeric DNA structures correlates with thermal stability of DNA duplexes.
Human DNA apurinic/apyrimidinic endonuclease 1 (APE1) is involved in the DNA base excision repair process. In addition to its AP (apurinic/apyrimidinic) endonucleolytic function, APE1 possesses 3' phosphodiesterase and 3'-5' exonuclease activities. The 3'-5' exonuclease activity is considered important in proofreading of DNA synthesis catalyzed by DNA polymerase beta. Here, we examine the removal of matched and mismatched dNMP from the 3' terminus of the 3'-recessed and nicked DNA by the APE1 activity using two different reaction buffers. To investigate whether the ability of APE1 to excise nucleotides from the 3' terminus depends on the thermal stability of the DNA duplex, we studied this characteristic of the DNAs that were used in the exonuclease assays in these two buffers. Our data confirm that APE1 removes mismatched nucleotides from the 3' terminus of DNA more efficiently than matched pairs. Both the efficiency of the 3'-5' exonuclease activity of APE1 and the thermal stability of DNA duplexes varied depending on the nature of the flanking group at the 5' margin of the nick. The 3'-5' exonuclease activity of APE1 shows a preference for substrates with a hydroxyl group at the 5' margin of the nick as well as for flapped and recessed DNAs.


Potent plasmodicidal activity of a heat-induced reformulation of deoxycholate-amphotericin B (Fungizone) against Plasmodium falciparum.
The emergence and spread of drug-resistant Plasmodium falciparum continue to pose problems in malaria chemotherapy. Therefore, it is necessary to identify new antimalarial drugs and therapeutic strategies. In the present study, the activity of a heat-treated form of amphotericin B (HT-AMB) against P. falciparum was evaluated. The efficacy and toxicity of HT-AMB were also compared with those of the standard formulation (AMB). HT-AMB showed significant activity against a chloroquine-resistant strain (strain K-1) and a chloroquine-susceptible strain (strain FCR-3) in vitro. The 50% inhibitory concentrations of HT-AMB were 0.32 +/- 0.03 mug/ml for strain K-1 and 0.33 +/- 0.03 mug/ml for strain FCR-3. In the presence of 1.0 mug of HT-AMB per ml, only pyknotic parasites were observed after 24 h of incubation of early trophozoites (ring forms). However, when late trophozoites and schizonts were cultured with 1.0 mug of HT-AMB per ml, those forms multiplied to ring forms but the number of infected erythrocytes did not increase. These results indicate that HT-AMB possesses potent antiplasmodial activity and that the drug is more effective against the ring-form stage than against the late trophozoite and schizont stages. HT-AMB was observed to have little cytotoxic effect against a human liver cell line (Chang liver cells). In conclusion, the results suggest that HT-AMB has promising properties and merits further in vivo investigations as a treatment for falciparum malaria.


G-protein-coupled receptor S1P1 acts within endothelial cells to regulate vascular maturation.
Sphingosine-1-phosphate (S1P) stimulates signaling pathways via G-protein-coupled receptors and triggers diverse cellular processes, including growth, survival, and migration. In S1P1 receptor-deficient embryos, blood vessels were incompletely covered by vascular smooth muscle cells (VSMCs), indicating the S1P1 receptor regulates vascular maturation. Because S1P1 receptor expression is not restricted to a particular cell type, it was not known whether the S1P1 receptor controlled VSMC coverage of vessels in a cell-autonomous fashion by functioning directly in VSMCs or indirectly through its activity in endothelial cells (ECs). By using the Cre/loxP system, we disrupted the S1P1 gene solely in ECs. The phenotype of the conditional mutant embryos mimicked the one obtained in the embryos globally deficient in S1P1. Thus, vessel coverage by VSMCs is directed by the activity of the S1P1 receptor in ECs.


Deletion of ferroportin in murine myeloid cells increases iron accumulation and stimulates osteoclastogenesis in vitro and in vivo.
Osteoporosis, osteopenia, and pathological bone fractures are frequent complications of iron-overload conditions such as hereditary hemochromatosis, thalassemia, and sickle cell disease. Moreover, animal models of iron overload have revealed increased bone resorption and decreased bone formation. Although systemic iron overload affects multiple organs and tissues, leading to significant changes on bone modeling and remodeling, the cell autonomous effects of excessive iron on bone cells remain unknown. Here, to elucidate the role of cellular iron homeostasis in osteoclasts, we generated two mouse strains in which solute carrier family 40 member 1 (Slc40a1), a gene encoding ferroportin (FPN), the sole iron exporter in mammalian cells, was specifically deleted in myeloid osteoclast precursors or mature cells. The FPN deletion mildly increased iron levels in both precursor and mature osteoclasts, and its loss in precursors, but not in mature cells, increased osteoclastogenesis and decreased bone mass in vivo Of note, these phenotypes were more pronounced in female than in male mice. In vitro studies revealed that the elevated intracellular iron promoted macrophage proliferation and amplified expression of nuclear factor of activated T cells 1 (Nfatc1) and PPARG coactivator 1β (Pgc-1β), two transcription factors critical for osteoclast differentiation. However, the iron excess did not affect osteoclast survival. While increased iron stimulated global mitochondrial metabolism in osteoclast precursors, it had little influence on mitochondrial mass and reactive oxygen species production. These results indicate that FPN-regulated intracellular iron levels are critical for mitochondrial metabolism, osteoclastogenesis, and skeletal homeostasis in mice.


Transtubular potassium gradient predicts kidney function impairment after adrenalectomy in primary aldosteronism.
In primary aldosteronism (PA), kidney function impairment could be concealed by relative hyperfiltration and emerge after adrenalectomy. We hypothesized transtubular gradient potassium gradient (TTKG), a kidney aldosterone bioactivity indicator, could correlate to end organ damage and forecast kidney function impairment after adrenalectomy. In the present prospective study, we enrolled lateralized PA patients who underwent adrenalectomy and were followed up 12 months after operation in the Taiwan Primary Aldosteronism Investigation (TAIPAI) registry from 2010 to 2018. The clinical outcome was kidney function impairment, defined as estimated glomerular filtration rate (eGFR) <60 ml/min/1.73 m2 at 12 months after adrenalectomy. End organ damage is determined by microalbuminuria and left ventricular mass. In total, 323 patients [mean, 50.8 ± 10.9 years old; female 178 (55.1%)] were enrolled. Comparing pre-operation and post-operation data, systolic blood pressure, serum aldosterone, urinary albumin to creatinine ratio and eGFR decreased. TTKG ⩾ 4.9 correlated with pre-operative urinary albumin to creatinine ratio >50 mg/g [odds ratio (OR) = 2.42; p = 0.034] and left ventricular mass (B = 20.10; p = 0.018). Multivariate logistic regression analysis demonstrated that TTKG ⩾ 4.9 could predict concealed chronic kidney disease (OR = 5.42; p = 0.011) and clinical success (OR = 2.90, p = 0.017) at 12 months after adrenalectomy. TTKG could predict concealed kidney function impairment and cure of hypertension in PA patients after adrenalectomy. TTKG more than 4.9 as an adverse surrogate of aldosterone and hypokalaemia correlated with pre-operative end organ damage in terms of high proteinuria and cardiac hypertrophy.


Amplified env and gag products on AKR cells. Origin from different murine leukemia virus genomes.
Thymocytes of AKR mice express two species of gp70, the envelope glycoprotein of murine leukemia virus (MuLV), encoded by the env gene. One is denoted Ec+ gp70 in reference to the type-antigen Ec and association with ecotropic virus. The other, Ec- gp70, resembles gp70 found also on thymocytes of mouse strains that are not overt producers of MuLV, and has no evident relation to ecotropic virus. Expression of Ec- gp70 type, but not of Ec+ gp70 type, is amplified with age on AKR thymocytes. In contrast, viral core polyproteins, encoded by the gag gene and simultaneously amplified with age, appear to be related to ecotropic virus. These observations imply selective amplification of products of env and gag genes from two sorts of provirus, a phenomenon which may be connected to the dual genetic origin of recombinant mink-cell-focus inducing viruses in AKR mice.


Genomic surveillance uncovers a pandemic clonal lineage of the wheat blast fungus.
Wheat, one of the most important food crops, is threatened by a blast disease pandemic. Here, we show that a clonal lineage of the wheat blast fungus recently spread to Asia and Africa following two independent introductions from South America. Through a combination of genome analyses and laboratory experiments, we show that the decade-old blast pandemic lineage can be controlled by the Rmg8 disease resistance gene and is sensitive to strobilurin fungicides. However, we also highlight the potential of the pandemic clone to evolve fungicide-insensitive variants and sexually recombine with African lineages. This underscores the urgent need for genomic surveillance to track and mitigate the spread of wheat blast outside of South America and to guide preemptive wheat breeding for blast resistance.


Renal function of substance P in rats chronically exposed to hypoxia.
We studied the renal action of substance P (SP) in rats chronically exposed to hypoxia (high altitude, HA), compared to control rats kept at sea level (SL). Hypoxia was induced by placing female Wistar rats (182-225 g) in an altitude chamber (5500 m) 15 h.d-1 for 4 weeks. Intrarenal arterial infusion of substance P (60 ng.kg-1. h-1) increased the excretion of urine, sodium in both groups of rats, however, the excretion of kallikrein (KK) and the glomerular filtration rate (GFR) were not significantly altered. After aprotinin (10,000 kiu.kg-1. min-1) treatment, kallikrein was depleted, and substance P lost its diuretic action. Spantide, an SP antagonist (6000 ng.kg-1. h-1, I.V.) decreased urine, and urinary excretion of sodium and potassium in SL rats, but not in HA rats. Acute renal denervated diuresis in SL rats was not modified after Spantide administration. Finally, it was found that both SP and SP antagonist did not significantly change the renal parameters in either group of rats after chronic renal denervation. We made the following conclusions: a) endogenous renal action of SP was suggested in SL but not in HA rats; b) the renal action of SP might be through KK release, although urinary KK did not increase after SP administration; and c) SP action is renal nerve dependent in both groups of rats.


Fabrication of metalosomes (metal containing cationic liposomes) using single chain surfactants as a precursor via formation of inorganic organic hybrids.
Among the self-assembled forms of surfactants, vesicles/liposomes are a highly promising and interesting feature of surfactants, which are usually formed from water insoluble surfactants. Herein, we demonstrate the formation of liposomes from single-chain cationic surfactants with the help of metals as a part of the counter ion, and these metal embedded liposomes are termed as metalosomes. It is a noteworthy advancement in the area of self-assembled molecular structures since we report the preparation of metal embedded liposomes (metalosomes) from a water soluble single chain cationic surfactant, which is otherwise a property or an arrangement made by double tailed surfactants, or more precisely lipids that are poorly water soluble. We can use this method for various cationic surfactants and metal combinations and the studies are still in process. However, this preliminary report on manganese-based surfactants depicts the successful formation of cationic metalosomes (with/without cholesterol), and the formation, structure and size has been verified using TEM, FE-SEM, DLS XRD and SAXS. The comparison of metalosomes with reverse vesicles in different solvents further gave an insight of microstructure and solvent environment effects on the self-assembly of metallosurfactants. In addition, we have also evaluated the encapsulation ability of metalosomes for fluorescein dye. High encapsulation efficiency of Mn-somes makes them promising candidates for several applications, particularly because of its water solubility.


Factors driving deforestation in common-pool resources in northern Mexico.
The theory of collective action has been extensively used to explain the relationship between common-based property regimes and the conservation of natural resources. However, there are two key components of the theory that literature reports as puzzles in which no consensus exists about their effect on the performance of common-pool resources. These are group size and heterogeneity. This study analyzes the effects of these two key components on the effectiveness of community-based forestry, called ejidos, to protect their forest resources in northern Mexico. We used a multinomial logit model to determine the contribution of 16 explanatory variables to the dependent variable, a measure of success of ejidos defined by the presence of deforested, degraded, or forested conditions. The results show that corn yield, marginality, percent of forest area, total population, a forest value index, distance to markets, roads and towns, were all statistically significant in driving deforested conditions. Deforestation becomes more attractive for poor communities and as corn yield and distance to towns, roads, and markets decrease. In general, group size and heterogeneity had no significant effects on the presence of deforested conditions. Deforestation is driven by resource-specific characteristics, such as location and soil productivity, not by ejidos' attributes, such as total area or number of members. We argue that current institutional policies focusing on the structure of property right arrangements should be shifted (1) to provide better technology for land cultivation; (2) to reduce the marginality problem in poor communities; and (3) to strengthen local institutions.


New insights into dissemination and variation of the health care-associated pathogen Acinetobacter baumannii from genomic analysis.
Acinetobacter baumannii is a globally important nosocomial pathogen characterized by an increasing incidence of multidrug resistance. Routes of dissemination and gene flow among health care facilities are poorly resolved and are important for understanding the epidemiology of A. baumannii, minimizing disease transmission, and improving patient outcomes. We used whole-genome sequencing to assess diversity and genome dynamics in 49 isolates from one United States hospital system during one year from 2007 to 2008. Core single-nucleotide-variant-based phylogenetic analysis revealed multiple founder strains and multiple independent strains recovered from the same patient yet was insufficient to fully resolve strain relationships, where gene content and insertion sequence patterns added additional discriminatory power. Gene content comparisons illustrated extensive and redundant antibiotic resistance gene carriage and direct evidence of gene transfer, recombination, gene loss, and mutation. Evidence of barriers to gene flow among hospital components was not found, suggesting complex mixing of strains and a large reservoir of A. baumannii strains capable of colonizing patients. Genome sequencing was used to characterize multidrug-resistant Acinetobacter baumannii strains from one United States hospital system during a 1-year period to better understand how A. baumannii strains that cause infection are related to one another. Extensive variation in gene content was found, even among strains that were very closely related phylogenetically and epidemiologically. Several mechanisms contributed to this diversity, including transfer of mobile genetic elements, mobilization of insertion sequences, insertion sequence-mediated deletions, and genome-wide homologous recombination. Variation in gene content, however, lacked clear spatial or temporal patterns, suggesting a diverse pool of circulating strains with considerable interaction between strains and hospital locations. Widespread genetic variation among strains from the same hospital and even the same patient, particularly involving antibiotic resistance genes, reinforces the need for molecular diagnostic testing and genomic analysis to determine resistance profiles, rather than a reliance primarily on strain typing and antimicrobial resistance phenotypes for epidemiological studies.


Acute Otitis Media and Other Complications of Viral Respiratory Infection.
Viral upper and lower respiratory tract infections (URI, LRI) are common in infants. We determined the prevalence of viral URI and its complications, including acute otitis media (AOM) and LRI, and assessed the effect of bacterial-viral interactions, and genetic and environmental risks on AOM development. Healthy infants were enrolled from near birth and followed to the first episode of AOM up to 12 months of age. Nasopharyngeal specimens were collected at monthly intervals (months 1-6, 9) and during viral URI episodes for bacterial culture and viral polymerase chain reaction studies. Subjects were followed closely for AOM development. A total of 367 infants were followed for 286 child-years; 887 URI (305 infants) and 180 AOM episodes (143 infants) were documented. Prevalence of URI, LRI, and AOM in the first year was 3.2, 0.25, and 0.67 per child-year, respectively. Cumulative AOM incidence by ages 3, 6, and 12 months was 6%, 23%, and 46%. Infants with and without AOM had 4.7 and 2.3 URI episodes per child-year, respectively (P < .002). Pathogenic bacterial colonization rates by month were significantly higher in infants with AOM (P < .005). Breastfeeding reduced both URI and AOM risks (P < .05). Significant bacterial-viral interactions occurred with Moraxella catarrhalis and a variety of respiratory viruses and altered URI and AOM risks. Almost half of infants experienced AOM by age 1. Important AOM risk factors included frequent viral URI, pathogenic bacterial colonization, and lack of breastfeeding. Bacterial-viral interactions may play a significant role in AOM pathogenesis and deserve further investigation.


Relationship of Terminal Duct Lobular Unit Involution of the Breast with Area and Volume Mammographic Densities.
Elevated mammographic density (MD) is an established breast cancer risk factor. Reduced involution of terminal duct lobular units (TDLU), the histologic source of most breast cancers, has been associated with higher MD and breast cancer risk. We investigated relationships of TDLU involution with area and volumetric MD, measured throughout the breast and surrounding biopsy targets (perilesional). Three measures inversely related to TDLU involution (TDLU count/mm(2), median TDLU span, median acini count/TDLU) assessed in benign diagnostic biopsies from 348 women, ages 40-65, were related to MD area (quantified with thresholding software) and volume (assessed with a density phantom) by analysis of covariance, stratified by menopausal status and adjusted for confounders. Among premenopausal women, TDLU count was directly associated with percent perilesional MD (P trend = 0.03), but not with absolute dense area/volume. Greater TDLU span was associated with elevated percent dense area/volume (P trend<0.05) and absolute perilesional MD (P = 0.003). Acini count was directly associated with absolute perilesional MD (P = 0.02). Greater TDLU involution (all metrics) was associated with increased nondense area/volume (P trend ≤ 0.04). Among postmenopausal women, TDLU measures were not significantly associated with MD. Among premenopausal women, reduced TDLU involution was associated with higher area and volumetric MD, particularly in perilesional parenchyma. Data indicating that TDLU involution and MD are correlated markers of breast cancer risk suggest that associations of MD with breast cancer may partly reflect amounts of at-risk epithelium. If confirmed, these results could suggest a prevention paradigm based on enhancing TDLU involution and monitoring efficacy by assessing MD reduction.


Killing and allowing to die in medical practice.
This paper examines some of the issues related to the distinction between acts and omissions. It discusses the difficulties involved in deciding whether there is any moral significance in this distinction, particularly when it is applied to cases which involve killing or allowing to die. The paper shows how this problem relates to some of the current issues in medical ethics. It examines the issues raised by the widely publicised cases of selective treatment of handicapped children and argues that such decisions are taken and have to be taken in the context of wider ethical theories.


[Effects of acute endurance exercise and 8 week training on the production of reactive oxygen species from neutrophils in untrained men].
We investigated the effects of acute endurance exercise and habitual physical activity for health maintenance on human neutrophil function in 12 untrained men. The acute exercise condition was a continuous exercise for 90 minutes at the intensity of 50% and 55% of maximal oxygen uptake (VO2max) on an ergometer. The training program was 3 km jogging three times per week for 8 weeks. The capacity of neutrophils to produce reactive oxygen species (ROS) was detected with lucigenin-dependent chemiluminescence (LgCL) and luminol-dependent chemiluminescence (LmCL) on stimulation with opsonized zymosan (OZ) and phorbol myristate acetate (PMA). As for the acute exercise effects, both LgCL and LmCL responses of neutrophils, stimulated using PMA consistently increased after exercise at 50%VO2max, whereas those stimulated with OZ remained unchanged. At 55%VO2max, LgCL responses to both stimulants increase maximally 1 h after exercise, and then decreased 3 h after exercise, whereas LmCL responses to both stimulants increased continuously after exercise at 55%VO2max. These phenomena observed at 55%VO2max compared to 50%VO2max suggests the improved capacity of producing ROS neutrophils after exercise. The number of neutrophils also increased maximally 1 h after exercise, due to the mobilization of band neutrophils (shift to the left), suggesting that functional changes was associated with cell mobilization. The increase in the capacity of neutrophils to produce ROS and marked neutrophilia following the acute endurance exercise suggests that a large quantity of ROS may be produced in vivo. As for the training effects, the LgCL and LmCL responses were maintained in the exercise group as compared to the decreased ones in the control group. The difference between the exercise group and the control group was observed only in LgCL response to OZ. Humoral immune factors (IgG, IgA, IgM, C3, C4) and serum opsonic activity were also unaltered. These phenomena suggest that homeostasis might be kept constant in terms of immunity through regular physical activity.


Phenotypic characterization of copper-resistant mutants of Methylosinus trichosporium OB3b.
Cultures of Methylosinus trichosporium OB3b grown in the presence of very low concentrations of copper synthesize a soluble methane monooxygenase (sMMO) that efficiently catalyzes the oxidation of trichloroethylene and other organic pollutants. Recently, we isolated five M. trichosporium OB3b mutants that express sMMO activity when grown in the presence of elevated copper concentrations (P.A. Phelps, S. K. Agarwal, G. E. Speitel, Jr., and G. Georgiou, Appl. Environ. Microbiol. 58:3701-3708, 1992). Here we show that, in contrast to the results for the wild-type cells, the addition of copper to mutant cultures grown on methane and nitrate as the nitrogen source has no noticeable effect on the growth rate and sMMO expression. In vitro experiments indicated that the copper-resistant phenotype does not arise from an increased stability of sMMO to copper deactivation. Furthermore, the mutant cultures exhibit altered speciation of copper in the extracellular fluid and have substantially decreased levels of cell-associated copper. On the basis of these results, we propose that the mutant phenotype arises from defects in copper uptake and metabolism rather than from changes in sMMO expression or enzyme stability.


Structural basis for the heterodimeric interaction between the acute leukaemia-associated transcription factors AML1 and CBFbeta.
Mutations in the genes encoding the interacting proteins AML1 and CBFbeta are the most common genetic abnormalities in acute leukaemia, and congenital mutations in the related AML3 gene are associated with disorders of osteogenesis. Furthermore, the interaction of AML1 with CBFbeta is essential for haematopoiesis. We report the 2.6 A resolution crystal structure of the complex between the AML1 Runt domain and CBFbeta, which represents a paradigm for the mode of interaction of this highly conserved family of transcription factors. The structure demonstrates that point mutations associated with cleidocranial dysplasia map to the conserved heterodimer interface, suggesting a role for CBFbeta in osteogenesis, and reveals a potential protein interaction platform composed of conserved negatively charged residues on the surface of CBFbeta.


Variable CD52 expression in mature T cell and NK cell malignancies: implications for alemtuzumab therapy.
The anti-CD52 antibody alemtuzumab has been explored as a novel targeted therapy in T cell malignancies. To assess the suitability of alemtuzumab therapy, we carried out a comprehensive study of CD52 expression using flow cytometry (FC) in 78 untreated patients diagnosed with mature T/natural killer (NK) cell neoplasms, including 34 adult T cell leukaemia/lymphomas (ATLL), two anaplastic large cell lymphomas (ALCL), three angioimmunoblastic T cell lymphomas (AITL), 16 cutaneous T cell lymphomas (CTCL), four extra-nodal T/NK cell lymphomas (ENT/NKCL), four hepatosplenic T cell lymphomas (HSTCL), 13 peripheral T cell lymphomas, not otherwise specified (PTCL-NOS) and two T-prolymphocytic leukaemia (T-PLL). The level of CD52 expression was quantified using QuantiBRITE standard beads. The level of CD52 expression varied widely within each diagnostic category. All AITL, HSTCL and T-PLL cases were CD52-positive and the frequency of CD52 expression was high in PTCL-NOS (92.3%), ATLL (94.1%) and CTCL (87.5%), implying a rational role for alemtuzumab in the treatment of these diseases; however, CD52 expression was low in ALCL (50%) and ENT/NKCL (25%). FC testing for cell surface expression of CD52 is indicated in patients with T/NK cell malignancies being considered for alemtuzumab therapy. Further studies are necessary to determine if the level of CD52 expression correlates with response to therapy.


Identification of VPS13C as a Galectin-12-Binding Protein That Regulates Galectin-12 Protein Stability and Adipogenesis.
Galectin-12, a member of the galectin family of β-galactoside-binding animal lectins, is preferentially expressed in adipocytes and required for adipocyte differentiation in vitro. This protein was recently found to regulate lipolysis, whole body adiposity, and glucose homeostasis in vivo. Here we identify VPS13C, a member of the VPS13 family of vacuolar protein sorting-associated proteins highly conserved throughout eukaryotic evolution, as a major galectin-12-binding protein. VPS13C is upregulated during adipocyte differentiation, and is required for galectin-12 protein stability. Knockdown of Vps13c markedly reduces the steady-state levels of galectin-12 by promoting its degradation through primarily the lysosomal pathway, and impairs adipocyte differentiation. Our studies also suggest that VPS13C may have a broader role in protein quality control. The regulation of galectin-12 stability by VPS13C could potentially be exploited for therapeutic intervention of obesity and related metabolic diseases.


Porin activity of the native and recombinant outer membrane protein Oms28 of Borrelia burgdorferi.
The outer membrane-spanning (Oms) proteins of Borrelia burgdorferi have been visualized by freeze-fracture analysis but, until recently, not further characterized. We developed a method for the isolation of B. burgdorferi outer membrane vesicles and described porin activities with single-channel conductances of 0.6 and 12.6 nS in 1 M KCI. By using both nondenaturing isoelectric focusing gel electrophoresis and fast-performance liquid chromatography separation after detergent solubilization, we found that the 0.6-nS porin activity resided in a 28-kDa protein, designated Oms28. The oms28 gene was cloned, and its nucleotide sequence was determined. The deduced amino acid sequence of Oms28 predicted a 257-amino-acid precursor protein with a putative 24-amino-acid leader peptidase I signal sequence. Processed Oms28 yielded a mature protein with a predicted molecular mass of 25,363 Da. When overproduced in Escherichia coli, the Oms28 porin fractionated in part to the outer membrane. Sodium dodecyl sulfate-polyacrylamide gel-purified recombinant Oms28 from E. coli retained functional activity as demonstrated by an average single-channel conductance of 1.1 nS in the planar lipid bilayer assay. These findings confirmed that Oms28 is a B. burgdorferi porin, the first to be described. As such, it is potential relevance to the pathogenesis of Lyme borreliosis and to the physiology of the spirochete.


Self-reported Vision Impairment and Subjective Well-being in Older Adults: A Longitudinal Mediation Analysis.
Vision impairment (VI) in older adults is associated with declines in well-being. However, the pathways through which poor vision leads to declines in well-being have not been well-described. The purpose of this study was to determine whether activity limitations and social participation restrictions mediate the impact of self-reported VI on subjective well-being. The National Health and Aging Trends Study (NHATS) is a nationally representative longitudinal study of Medicare beneficiaries 65 and older that includes detailed measures of the disablement process. A longitudinal mediation model was conceptualized linking self-reported VI and subjective well-being. Structural equation modeling was used to test the mediating effects of activity limitations and social participation restrictions while adjusting for relevant covariates. The final sample included 5,431 respondents. At baseline, 8.0% of Medicare beneficiaries had self-reported VI. Subjective well-being scores were significantly lower among respondents with self-reported VI (15.7; 95% confidence interval [CI]: 15.2, 16.2) compared with those without VI (17.6; 95% CI: 17.5, 17.7). Self-reported VI had a significant indirect effect on subjective well-being through limiting mobility (β = -0.04; 95% CI: -0.07, -0.03) and household activities (β = -0.05; 95% CI: -0.08, -0.03), but not self-care limitations (β = 0.0; 95% CI = 0.0, 0.0) or participation restrictions (β = 0.0; 95% CI = -0.01, 0.00). Total indirect effects from all mediation paths accounted for 42% of the effect of VI on well-being. Mobility and household activity limitations are significant mediators that explain a considerable portion of the impact of poor vision on well-being. Interventions to promote successful accommodation may result in greater overall well-being for older adults with poor vision.


Phase of Care Mortality Analysis: A Unique Method for Comparing Mortality Differences Among Transcatheter Aortic Valve Replacement and Surgical Aortic Valve Replacement Patients.
The objective of this study is based on the phase of care mortality analysis (POCMA), an effective tool to evaluate the root cause of in-hospital mortality in cardiac surgery patients. POCMA has not been used to compare operative mortalities among transcatheter aortic valve replacement (TAVR) and surgical aortic valve replacement (SAVR) populations, and may provide insight that could affect patient safety initiatives and improve outcomes in aortic valve surgery. We included patients who underwent TAVR or isolated SAVR between 2011 and March 31, 2015 and did not survive the index hospitalization. A multidisciplinary heart team made POCMA assignments as part of the weekly morbidity and mortality conference, pinpointing the phase of care and subcategory that directly caused or had the greatest effect on each mortality. During the study period, 240 patients underwent TAVR and 530 underwent SAVR. Unadjusted mortality rates were significantly higher in the TAVR group, 5.0% (n = 12) compared with SAVR, 1.9% (n = 10) (P = 0.016). TAVR deaths by phase of care are as follows: 0 for preoperative, 9 (72.8%) for intraoperative, 2 (18.2%) for postoperative intensive care unit, and 1 (9.1%) for postoperative floor. By comparison, 4 (40%) SAVR deaths had a root cause in the preoperative phase, 1 (10%) in the intraoperative phase, and 5 (50%) in the postoperative intensive care unit phase. POCMA is a novel method of categorizing in-hospital mortalities. Our single institution review revealed that patients who underwent TAVR more often expired because of intraoperative technical issues, whereas SAVR deaths were typically the result of patient selection or postoperative complications.


First Report of Johnsongrass mosaic virus (JGMV) Infecting Pennisetum purpureum in Brazil.
Tropical grass and legume species used as pasture grasses for cattle feeding cover over 25% of the agricultural area in Brazil. In recent years, plants showing virus-like symptoms have been observed in the main pasture grass growing areas. Plants of Pennisetum purpureum line CNPGL 00211 showing typical virus mosaic symptoms on leaves and growth reduction were collected in Bahia State, Brazil. Flexuous elongated potyvirus-like particles were observed in the leaf-dip preparation of diseased plants by electron microscopy. In addition, the virus was mechanically transmitted using a standard procedure for potyviruses (4) and produced similar symptoms in inoculated P. purpureum plants. For further molecular identification, total RNA was extracted from frozen symptomatic leaves following the guanidine thiocyanate method (3). cDNA synthesis was performed using oligonucleotide, OligodT50M10 and PCR was carried out using Potyvirus degenerate primers PY11 (5'-GGNAAYAAYAGYGGNCARCC-3') (2) and M10 (5'-AAGCAGTGTTATCAACGCAGA-3'). The amplified fragments of the expected size (approximately 2 kb comprising part of the NIb protein gene, the entire coat protein [CP] gene, and the 3' nontranslated region) were separated using agarose gel electrophoresis, excised, and cloned into plasmid vector pGEMT-Easy (Promega) according to the manufacturer's instructions. Four selected clones were sequenced (Macrogen, South Korea). The sequenced 2.0-kb fragment (GenBank Accession No. KC333416) was compared with sequences available in GenBank and the highest nucleotide identity of 79% was observed with Johnsongrass mosaic virus (JGMV) isolated in Australia (4). According to the Potyvirus species demarcation convention based on CP identity (1), the virus isolate from P. purpureum belongs to the JGMV species. However, the amino acid sequence of the N-terminus of the CP of the Bahia isolate is distinct from JGMV sequences reported in GenBank. The phylogenetic analysis of the CP confirmed the difference since this Bahia isolate was located in a clearly distinct branch separate from all JGMV isolates. To our knowledge, this is the first report of a JGMV in Brazil infecting tropical grass in the main pasture areas. References: (1) M. J. Adams et al. Arch. Virol. 150: 459, 2005. (2) J. Chen et al. Arch. Virol. 146:757. 2001. (3) P. Chomczynski and N. Sacchi. Nature Protocols 1:581, 2006. (4) H. K. Laidlaw et al. Arch. Virol. 149:1633, 2004.


Working-point control method for readout of dynamic phase changes in interferometric fiber-optic sensors by tuning the laser frequency.
A simple but reliable method, namely the working-point control by tuning the laser frequency, for the dynamic phase shift measurement in a passive homodyne interferometric fiber-optic sensor is proposed. A dc voltage calculated from the photodetector output is applied to the light source to control the interferometer at the condition of maximum sensitivity. Then the signal's phase shift can be obtained from the components of zero and fundamental frequencies. To test the method, an all polarization-maintaining Mach-Zehnder interferometer with a piezoelectric ceramic (PbZrTiO(3), or PZT) cylinder in one arm is constructed. The experimental results show that the simulation signal's phase shift generated by the PZT cylinder can be read out correctly with the method. It has the advantages of simplicities of operation, no-active element in the sensing head, and large operating bandwidth. It can be used for readout of dynamic phase shifts in various interferometric fiber-optic sensors.


Investigation of ancient DNA from Western Siberia and the Sargat culture.
Mitochondrial DNA from 14 archaeological samples at the Ural State University in Yekaterinburg, Russia, was extracted to test the feasibility of ancient DNA work on their collection. These samples come from a number of sites that fall into two groupings. Seven samples are from three sites, dating to the 8th-12th century AD, that belong to a northern group of what are thought to be Ugrians, who lived along the Ural Mountains in northwestern Siberia. The remaining seven samples are from two sites that belong to a southern group representing the Sargat culture, dating between roughly the 5th century BC and the 5th century AD, from southwestern Siberia near the Ural Mountains and the present-day Kazakhstan border. The samples are derived from several burial types, including kurgan burials. They also represent a number of different skeletal elements and a range of observed preservation. The northern sites repeatedly failed to amplify after multiple extraction and amplification attempts, but the samples from the southern sites were successfully extracted and amplified. The sequences obtained from the southern sites support the hypothesis that the Sargat culture was a potential zone of intermixture between native Ugrian and/or Siberian populations and steppe peoples from the south, possibly early Iranian or Indo-Iranian, which has been previously suggested by archaeological analysis.


Unravelling the genetics of inherited retinal dystrophies: Past, present and future.
The identification of the genes underlying monogenic diseases has been of interest to clinicians and scientists for many years. Using inherited retinal dystrophies as an example of monogenic disease we describe the history of molecular genetic techniques that have been pivotal in the discovery of disease causing genes. The methods that were developed in the 1970's and 80's are still in use today but have been refined and improved. These techniques enabled the concept of the Human Genome Project to be envisaged and ultimately realised. When the successful conclusion of the project was announced in 2003 many new tools and, as importantly, many collaborations had been developed that facilitated a rapid identification of disease genes. In the post-human genome project era advances in computing power and the clever use of the properties of DNA replication has allowed the development of next-generation sequencing technologies. These methods have revolutionised the identification of disease genes because for the first time there is no need to define the position of the gene in the genome. The use of next generation sequencing in a diagnostic setting has allowed many more patients with an inherited retinal dystrophy to obtain a molecular diagnosis for their disease. The identification of novel genes that have a role in the development or maintenance of retinal function is opening up avenues of research which will lead to the development of new pharmacological and gene therapy approaches. Neither of which can be used unless the defective gene and protein is known. The continued development of sequencing technologies also holds great promise for the advent of truly personalised medicine.


Occurrence of maltreatment in active duty military and nonmilitary families in the State of Texas.
This study compared the occurrence of child maltreatment in military and nonmilitary populations. Data came from the National Child Abuse and Neglect Data System. All cases of child maltreatment substantiated from January 1, 2000 through December 31, 2002, in the state of Texas were analyzed. The rate of occurrence of substantiated maltreatment was lower for children in military families compared with children in nonmilitary families (5.05 vs. 7.89 per 1000 person-years). A greater proportion of caretakers of children in nonmilitary families compared with military families reportedly had financial problems (18.7% vs. 5.2%) and received public assistance (28.2% vs. 8.9%). The rate of occurrence of substantiated child maltreatment appears to be lower in military than nonmilitary families. The proportion of families with financial problems and public assistance is lower in military than nonmilitary families, suggesting that these factors may increase stress in families and the risk of maltreatment.


Sepsis carries a high mortality among hospitalised adults in Malawi in the era of antiretroviral therapy scale-up: a longitudinal cohort study.
To assess mortality risk among adults presenting to an African teaching hospital with sepsis and severe sepsis in a setting of high HIV prevalence and widespread ART uptake. Prospective cohort study of adults (age ≥16 years) admitted with clinical suspicion of severe infection between November 2008 and January 2009 to Queen Elizabeth Central Hospital, a 1250-bed government-funded hospital in Blantyre, Malawi. Demographic, clinical and laboratory information, including blood and cerebrospinal fluid cultures were obtained on admission. Data from 213 patients (181 with sepsis and 32 with severe sepsis; M:F = 2:3) were analysed. 161 (75.6%) patients were HIV-positive. Overall mortality was 22%, rising to 50% amongst patients with severe sepsis. The mortality of all sepsis patients commenced on antiretroviral therapy (ART) within 90 days was 11/28 (39.3%) compared with 7/42 (16.7%) among all sepsis patients on ART for greater than 90 days (p = 0.050). Independent associations with death were hypoxia (OR = 2.4; 95% CI, 1.1-5.1) and systolic hypotension (OR 7.0; 95% CI: 2.4-20.4). Sepsis and severe sepsis carry high mortality among hospitalised adults in Malawi. Measures to reduce this, including early identification and targeted intervention in high-risk patients, especially HIV-positive individuals recently commenced on ART, are urgently required.


Nitrogen and phosphorus attenuation within the stream network of a coastal, agricultural watershed.
Streams alter the concentration of nutrients they transport and thereby influence nutrient loading to estuaries downstream; however, the relationship between in-stream uptake, discharge variability, and subsequent nutrient export is poorly understood. In this study, in-stream N and P uptake were examined in the stream network draining a row-crop agricultural operation in coastal North Carolina. The effect of in-stream nutrient uptake on estuarine loading was examined using continuous measurements of watershed nutrient export. From August to December 2003, 52 and 83% of the NH4+ and PO4(3-) loads were exported during storms while concurrent storm flow volume was 34% of the total. Whole-ecosystem mass transfer velocities (Vf) of NH4+ and PO4(3-), measured using short-term additions of inorganic nutrients, ranged from 0.1 to 25 mm min(-1). Using a mass balance approach, this in-stream uptake was found to attenuate 65 to 98% of the NH4+ flux and 78 to 98% of the PO4(3-) flux in small, first-order drainage ditches. For the larger channel downstream, an empirical model based on Vf and discharge was developed to estimate the percentage of the nutrient load retained in-stream. The model predicted that all of the upstream NH4+ and PO4(3-) load was retained during base flow, while 65 and 37% of the NH4+ and PO4(3-) load was retained during storms. Remineralization from the streambed (vs. terrestrial sources) was the apparent source of NH4+ and PO4(3-) to the estuary during base flow. In-stream uptake reduced the dissolved inorganic N to dissolved inorganic P ratio of water exported to the N-limited estuary, thus limiting the potential for estuarine phytoplankton growth.


Stigma as a barrier to family planning use among married youth in Ethiopia.
Nearly 33 million female youths have an unmet need for voluntary family planning (FP), meaning they are sexually active and do not want to become pregnant. In Ethiopia, age at marriage remains low: 40% and 14% of young women aged 20-24 were married by the ages of 18 and 15, respectively. Despite increases in FP use by married 15- to 24-year-olds from 5% in 2000 to 37% in 2016, unmet need remains high at 19%. Supply-and-demand factors have been shown to limit FP use, yet little is known about how stigma influences FP use among youth. This study validates an anticipated stigma (expectation of discrimination from others) index and explores its effect on unmet need. A cross-sectional survey was implemented with 15- to 24-year-old female youth in Ethiopia in 2016. The analytic sample included married respondents with a demand (met and unmet need) for FP (n=371). A five-item anticipated stigma index (Cronbach's α=0.66) was developed using principal component factor analysis. These items related to fear, worry and embarrassment when accessing FP. The findings showed that 30% agreed with at least one anticipated stigma question; 44% had an unmet need; 58% were married before age 18; and 100% could name an FP method and knew where to obtain FP. In multivariate regression models, youth who experienced anticipated stigma were significantly more likely to have an unmet need, and those who lived close to a youth-friendly service (YFS) site were significantly less likely to have an unmet need. Interventions should address anticipated stigma while focusing on social norms that restrict married youth from accessing FP; unmet need may be mitigated in the presence of a YFS; and the anticipated stigma index appears valid and reliable but should be tested in other countries and among different adolescent groups.


Replication kinetics of coxsackievirus A16 in human rhabdomyosarcoma cells.
Coxsackievirus A16 (CVA16), together with enterovirus type 71 (EV71), is responsible for most cases of hand, foot and mouth disease (HFMD) worldwide. Recent findings suggest that the recombination between CVA16 and EV71, and the co-circulation of these two viruses may have contributed to the increase of HFMD cases in China over the past few years. It is therefore important to further understand the virology, epidemiology, virus-host interactions and host pathogenesis of CVA16. In this study, we describe the viral kinetics of CVA16 in human rhabdomyosarcoma (RD) cells by analyzing the cytopathic effect (CPE), viral RNA replication, viral protein expression, viral RNA package and viral particle secretion in RD cells. We show that CVA16 appears to first attach, uncoat and enter into the host cell after adsorption for 1 h. Later on, CVA16 undergoes rapid replication from 3 to 6 h at MOI 1 and until 9 h at MOI 0.1. At MOI 0.1, CVA16 initiates a secondary infection as the virions were secreted before 9 h p.i. CPE was observed after 12 h p.i., and viral antigen was first detected at 6 h p.i. at MOI 1 and at 9 h p.i. at MOI 0.1. Thus, our study provides important information for further investigation of CVA16 in order to better understand and ultimately control infections with this virus.


Anorexia nervosa in Hong Kong. Why not more in Chinese?
Anorexia nervosa is a geographically distinct psychiatric disorder; it is rapidly increasing in incidence in Western countries, while being virtually unreported in China, or in the Chinese community of Hong Kong. This is surprising when the Chinese preoccupation with food and their reported readiness to somatise dysphoria are considered. Three Chinese anorectics born and living in Hong Kong and exhibiting mostly typical clinical features are reported. The rarity of the disorder in the East could be related to protective biological and socio-cultural factors specific to the Chinese, and while it may become more common, anorexia nervosa is unlikely to reach Western proportions.


NMDARs in granule cells contribute to parallel fiber-Purkinje cell synaptic plasticity and motor learning.
Long-term synaptic plasticity is believed to be the cellular substrate of learning and memory. Synaptic plasticity rules are defined by the specific complement of receptors at the synapse and the associated downstream signaling mechanisms. In young rodents, at the cerebellar synapse between granule cells (GC) and Purkinje cells (PC), bidirectional plasticity is shaped by the balance between transcellular nitric oxide (NO) driven by presynaptic N-methyl-D-aspartate receptor (NMDAR) activation and postsynaptic calcium dynamics. However, the role and the location of NMDAR activation in these pathways is still debated in mature animals. Here, we show in adult rodents that NMDARs are present and functional in presynaptic terminals where their activation triggers NO signaling. In addition, we find that selective genetic deletion of presynaptic, but not postsynaptic, NMDARs prevents synaptic plasticity at parallel fiber-PC (PF-PC) synapses. Consistent with this finding, the selective deletion of GC NMDARs affects adaptation of the vestibulo-ocular reflex. Thus, NMDARs presynaptic to PCs are required for bidirectional synaptic plasticity and cerebellar motor learning.


Prospective, randomized comparison of 2 methods of cold snare polypectomy for small colorectal polyps.
Both cold-only snare and hot polypectomy snare are used for the removal of small colorectal polyps. To compare the outcome of cold snare polypectomy of small colorectal polyps with a snare exclusively designed as a cold snare versus cold snare polypectomy by using a traditional polypectomy snare. Prospective, randomized, controlled study. Municipal hospital in Japan. Patients with colorectal polyps 10 mm or smaller in diameter were randomized to dedicated cold snare (dedicated cold snare group) or traditional cold snare (traditional cold snare group). The primary outcome measure was complete resection rates by cold snaring based on pathological examination. Secondary outcomes included bleeding within 2 weeks after polypectomy and identification of submucosal arteries and injured arteries in the resected specimens. Seventy-six patients having 210 eligible polyps were randomized: dedicated cold snare group, N = 37 (98 polyps) and traditional cold snare group, N = 39 (112 polyps). Patient demographic characteristics including the number, size, and shape of the polyps removed were similar in the 2 groups. The complete resection rate was significantly greater with the dedicated cold than with the traditional cold snare (91% [89/98] vs 79% [88/112], P = .015), with a marked difference with 8- to 10-mm polyps, both flat and pedunculated. Immediate bleeding and hematochezia rates were similar (19% vs 21%, P = .86; 5.4% vs 7.7%, P = .69). No delayed bleeding occurred. Histology demonstrated a similar prevalence of arteries and injured arteries in the submucosa (33% [32/96] vs 30% [31/104], P = .59; 3.1% [3/96] vs 6.7% [7/104], P = .24). Small sample size, single-center study. Polypectomy by using a dedicated cold snare resulted in complete polyp removal more often than did cold snaring with a traditional snare, especially polyps 8 to 10 mm in diameter, whether flat or pedunculated. ( NCT02036047.)


Sensory organization for balance: specific deficits in Alzheimer's but not in Parkinson's disease.
The cause of frequent falling in patients with dementia of the Alzheimer type (AD) is not well understood. Distraction from incongruent visual stimuli may be an important factor as suggested by their poor performance in tests of shifting visual attention in other studies. The purpose of this study was to determine whether AD patients have difficulty maintaining upright balance under absent and/or incongruent visual and other sensory conditions compared to nondemented healthy elderly persons and individuals with Parkinson's disease (PD). Seventeen healthy older adults, 15 medicated PD subjects, and 11 AD subjects underwent the Sensory Organization Test protocol. The incidence of loss of balance ("falls"), and the peak-to-peak amplitude of body center of mass sway during stance in the six sensory conditions were used to infer the ability to use visual, somatosensory, and vestibular signals when they provided useful information for balance, and to suppress them when they were incongruent as an orientation reference. Vestibular reflex tests were conducted to ensure normal vestibular function in the subjects. AD subjects had normal vestibular function but had trouble using it in condition 6, where they had to concurrently suppress both incongruent visual and somatosensory inputs. All 11 AD subjects fell in the first trial of this condition. With repeated trials, only three AD subjects were able to stay balanced. AD subjects were able to keep their balance when only somatosensory input was incongruent. In this condition, all AD subjects were able to maintain balance whereas some falls occurred in the other groups. In all conditions, when AD subjects did not fall, they were able to control as large a sway as the healthy controls, except when standing with eyes closed in condition 2: AD subjects did not increase their sway whereas the other groups did. In the PD group, the total fall incidence was similar to the AD group, but the distribution was generalized across more sensory conditions. PD subjects were also able to improve with repeated trials in condition 6. Patients with dementia of the Alzheimer type have decreased ability to suppress incongruent visual stimuli when trying to maintain balance. However, they did not seem to be dependent on vision for balance because they did not increase their sway when vision was absent. Parkinsonian patients have a more general balance control problem in the sensory organization test, possibly related to difficulty changing set.


Pre-operative optimisation for chronic obstructive pulmonary disease: a narrative review.
Chronic obstructive pulmonary disease is a condition commonly present in older people undergoing surgery and confers an increased risk of postoperative complications and mortality. Although predominantly a respiratory disease, it frequently has extra-pulmonary manifestations and typically occurs in the context of other long-term conditions. Patients experience a range of symptoms that affect their quality of life, functional ability and clinical outcomes. In this review, we discuss the evidence for techniques to optimise the care of people with chronic obstructive pulmonary disease in the peri-operative period, and address potential new interventions to improve outcomes. The article centres on pulmonary rehabilitation, widely available for the treatment of stable chronic obstructive pulmonary disease, but less often used in a peri-operative setting. Current evidence is largely at high risk of bias, however. Before surgery it is important to ensure that what have been called the 'five fundamentals' of chronic obstructive pulmonary disease treatment are achieved: smoking cessation; pulmonary rehabilitation; vaccination; self-management; and identification and optimisation of co-morbidities. Pharmacological treatment should also be optimised, and some patients may benefit from lung volume reduction surgery. Psychological and behavioural factors are important, but are currently poorly understood in the peri-operative period. Considerations of the risk and benefits of delaying surgery to ensure the recommended measures are delivered depends on patient characteristics and the nature and urgency of the planned intervention.


How to decrease suicide rates in both genders? An effectiveness study of a community-based intervention (EAAD).
The suicide rate in Hungary is high in international comparison. The two-year community-based four-level intervention programme of the European Alliance Against Depression (EAAD) is designed to improve the care of depression and to prevent suicidal behaviour. Our aim was to evaluate the effectiveness of a regional community-based four-level suicide prevention programme on suicide rates. The EAAD programme was implemented in Szolnok (population 76,311), a town in a region of Hungary with an exceptionally high suicide rate. Effectiveness was assessed by comparing changes in suicide rates in the intervention region after the intervention started with changes in national suicide rates and those in a control region (Szeged) in the corresponding period. For the duration of the programme and the follow-up year, suicide rates in Szolnok were significantly lower than the average of the previous three years (p = .0076). The suicide rate thus went down from 30.1 per 100,000 in 2004 to 13.2 in 2005 (-56.1 %), 14.6 in 2006 (-51.4 %) and 12.0 in 2007 (-60.1 %). This decrease of annual suicide rates in Szolnok after the onset of the intervention was significantly stronger than that observed in the whole country (p = .017) and in the control region (p = .0015). Men had the same decrease in suicide rates as women. As secondary outcome, an increase of emergency calls to the hotline service (200%) and outpatient visits at the local psychiatry clinic (76%) was found. These results seem to provide further support for the effectiveness of the EAAD concept. Whilst the majority of suicide prevention programs mainly affect female suicidal behaviour, this programme seems to be beneficial for both sexes. The sustainability and the role of the mediating factors (social service and health care utilization, community attitudes about suicide) should be key points in future research.


Stimulatory effects of 4-hydroxytamoxifen on proliferation of human endometrial adenocarcinoma cells (Ishikawa line).
The effects of trans-4-hydroxytamoxifen (OHTam) on proliferation of cells of the Ishikawa human endometrial adenocarcinoma line were studied under serum-free, phenol red-free conditions and compared to those of estradiol. The addition of OHTam (1 microM) to basal medium (BM), consisting of equal parts of Dulbecco's modified Eagle's medium and Ham's F-12 with additional glutamine and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, resulted in significant increases in cell numbers relative to controls. These effects were even greater than those obtained with estradiol (10 nM-1 microM) or 1% charcoal-treated fetal bovine serum (ctFBS). Addition of 1% ctFBS to BM containing 1 microM OHTam further increased cell numbers whereas addition of estradiol (10 nM) did not do so. The stimulation of growth was positively correlated with OHTam concentrations in the range of 10 nM to 1 microM. Dissociation of estradiol and OHTam proliferative effects was observed in a variant of Ishikawa cells in which estradiol did not increase proliferation while OHTam had a strong stimulatory effect. The growth-promoting effects of OHTam were also observed in BM containing 5% or 15% ctFBS. In contrast, in parallel experiments in which BM was replaced by minimal essential medium (Eagle's) with Earle's salts, OHTam (1 microM) did not stimulate proliferation under these conditions and acted as an antiestrogen, inhibiting the proliferative effects of estradiol. These results illustrate marked effects of medium composition on proliferation and antiestrogenic actions of OHTam. Alkaline phosphatase activity was strongly stimulated by estradiol (10 nM) but only very weakly affected by OHTam (1 microM); at these concentrations, OHTam inhibited the effect of estradiol, both in serum-free BM and in minimal essential medium plus 15% ctFBS, demonstrating dissociation in its actions on proliferation and on enzymatic activity. These findings suggest that OHTam may stimulate the proliferation of particular clones of endometrial cancer cells in human tumors. They also suggest that OHTam can exert effects not mediated by the estrogen receptor system, or form OHTam-estrogen receptor agonistic complexes unlike those resulting from estradiol-estrogen receptor interactions. Clearly, Ishikawa cells provide a useful model to investigate mechanisms of action of antiestrogens.


Premature ovarian failure and dehydroepiandrosterone.
Since the first successful birth at our center following treatment with DHEA in a patient with premature ovarian failure (POF), all subsequent patients with POF underwent the same treatment protocol. Receiving very encouraging results (FSH level was decreased in the first five patients and all achieved pregnancy), it was decided to study the action of DHEA for a reasonable length of time and in a larger number of patients to confirm the effectiveness of this particular therapeutic regime.


Activity profiles of enzymes that control the uracil incorporation into DNA during neuronal development.
We have shown that DNA polymerase beta, the only nuclear DNA polymerase present in adult neurons, cannot discriminate between dTTP and dUTP, having the same Km for both substrates. This fact suggests that during reparative DNA synthesis, in adult neurons, dUMP residues can be incorporated into DNA. Since uracil DNA-glycosylase functions to prevent the mutagenic effects of uracil in DNA coming as a product of deamination of cytosine residues or as a result of dUMP incorporation by DNA polymerase, we have studied the perinatal activity of uracil DNA-glycosylase and of 2 enzymes (nucleoside diphosphokinase and dUTPase) involved in dUTP metabolism. Our data indicate that during neuronal development there is a rapid decrease in uracil DNA-glycosylase which could impair the removal of uracil present in DNA in adult neurons. However, misincorporation of dUMP into DNA might be kept to a low frequency by the action of dUTPase present at all developmental stages.


Magnetic resonance imaging of suspicious breast masses seen on one mammographic view.
The purpose of this study was to assess the utility of contrast-enhanced breast magnetic resonance imaging (MRI) in identifying lesions unidentified on the craniocaudal projection. The authors reviewed five patients with suspicious mammographic lesions not imaged on the craniocaudal mammogram who were referred for contrast-enhanced MRI and underwent subsequent preoperative needle localization in four of the five cases. Five patients, ages 56 to 69 years, had suspicious lesions identified on mediolateral oblique (MLO) or mediolateral (ML) projections only. Ultrasound did not identify the lesion in any of these cases. MRI identified suspicious breast lesions measuring 5 to 12 mm in size. These were located high on the chest wall or in the upper inner quadrant. Suspicious lesions seen only on the MLO or ML projections may reside high on the chest wall or in the upper inner quadrant. Lesions in these locations may be typically excluded on the craniocaudal projection during mammography. Breast MRI has the advantage of imaging the entire breast and is particularly useful for these lesions. In this series, MRI prevented delay in breast cancer diagnosis.


EGFR signalling as a negative regulator of Notch1 gene transcription and function in proliferating keratinocytes and cancer.
The Notch1 gene has an important role in mammalian cell-fate decision and tumorigenesis. Upstream control mechanisms for transcription of this gene are still poorly understood. In a chemical genetics screen for small molecule activators of Notch signalling, we identified epidermal growth factor receptor (EGFR) as a key negative regulator of Notch1 gene expression in primary human keratinocytes, intact epidermis and skin squamous cell carcinomas (SCCs). The underlying mechanism for negative control of the Notch1 gene in human cells, as well as in a mouse model of EGFR-dependent skin carcinogenesis, involves transcriptional suppression of p53 by the EGFR effector c-Jun. Suppression of Notch signalling in cancer cells counteracts the differentiation-inducing effects of EGFR inhibitors while, at the same time, synergizing with these compounds in induction of apoptosis. Thus, our data reveal a key role of EGFR signalling in the negative regulation of Notch1 gene transcription, of potential relevance for combinatory approaches for cancer therapy.


Correlation of PP and PR intervals in premature low birth weight infants.
We examined the developmental change by which autonomic neural activity associated respiration modulates spontaneous firing rate of sinus (SA) node and atrioventricular (AV) conduction in premature infants born with low birth weight (LBWI). The purpose of this study was to clarify whether variation of PR is correlated with that of PP or those are independent in LBWI with immature autonomic nervous system. We investigated, therefore, whether there are spontaneous functional differences in the innervation of SA and AV nodes. Further, we evaluated the maturation of autonomic nervous system progressing in the period, on the day of birth (Day 0) to approximately one month after the birth (Month 1). This study was performed in thirteen LBWI during deep sleep. EEG, EOG, ECG, respiratory waves were digitized on line, spontaneous firing cycle of SA node (PP), and AV nodal conduction time (PR) that were recorded on Day 0 and Month 1. Then, the data were analyzed as follows: 1) correlations among the means and standard deviations (SD) of PP, PR and RR, 2) variance evaluation of PP and PR intervals by Lorenz plot analysis method, 3) correlation analysis among PP, PR and RR intervals by linear regression method and 4) frequency analysis for PP and PR intervals by high-speed Fourier transform method (FFT) and determination of frequency density. The PP interval decreased as growing in the period. Contrary PR interval increased. In LBWI, the automatic nervous activities including parasympathetic nerve activity for spontaneous firing cycle of SA node and ventricular excitation cycle on Month 1 were higher than Day 0. It was assumed that the vagal nerve activity for the AV conduction was enhanced. However, there was no significant change in linear regression slope for the spontaneous firing cycle of SA node and the AV conduction time. Postnatal LF/HF changes for PP and PR obtained by frequency analysis, were opposite. Therefore, it was suggested that the maturity of autonomic nervous system progresses in the period, Day 0 to approximately Month 1, but the variations in PP and PR are independent each other.


Smartphone-based detection of unlabeled DNA via electrochemical dissolution.
We describe a novel approach that enables unlabeled biomolecules and chemical analytes to be detected using ordinary smartphone optics. Electrochemical reactivity of chromium, ordinarily considered detrimental, is harnessed here to generate a signature that can be easily seen by monitoring electrode dissolution under ordinary white-light illumination. The simplicity and robustness of this approach eliminates the need for labeling and/or pre-programming with specific receptors (e.g., oligonucleotide probes), making it feasible to greatly expand availability of a host of assays where detection complexity is a limiting constraint.


Evaluation at scale of microbiome-derived metabolites as biomarker of flavan-3-ol intake in epidemiological studies.
The accurate assessment of dietary intake is crucial to investigate the effect of diet on health. Currently used methods, relying on self-reporting and food composition data, are known to have limitations and might not be suitable to estimate the intake of many bioactive food components. An alternative are nutritional biomarkers, which can allow an unbiased assessment of intake. They require a careful evaluation of their suitability, including: (a) the availability of a precise, accurate and robust analytical method, (b) their specificity (c) a consistent relationship with actual intake. We have evaluated human metabolites of a microbiome-derived flavan-3-ol catabolite, 5-(3',4'-dihydroxyphenyl)-[gamma]-valerolactone (gVL), as biomarker of flavan-3-ol intake in large epidemiological studies. Flavan-3-ols are widely consumed plant bioactives, which have received considerable interest due to their potential ability to reduce CVD risk. The availability of authentic standards allowed the development of a validated high-throughput method suitable for large-scale studies. In dietary intervention studies, we could show that gVL metabolites are specific for flavan-3-ols present in tea, fruits, wine and cocoa-derived products, with a strong correlation between intake and biomarker (Spearman's r = 0.90). This biomarker will allow for the first time to estimate flavan-3-ol intake and further investigation of associations between intake and disease risk.


Amplification of surface temperature trends and variability in the tropical atmosphere.
The month-to-month variability of tropical temperatures is larger in the troposphere than at Earth's surface. This amplification behavior is similar in a range of observations and climate model simulations and is consistent with basic theory. On multidecadal time scales, tropospheric amplification of surface warming is a robust feature of model simulations, but it occurs in only one observational data set. Other observations show weak, or even negative, amplification. These results suggest either that different physical mechanisms control amplification processes on monthly and decadal time scales, and models fail to capture such behavior; or (more plausibly) that residual errors in several observational data sets used here affect their representation of long-term trends.


The endothelin B receptor plays a crucial role in the adhesion of neutrophils to the endothelium in sickle cell disease.
Although the primary origin of sickle cell disease is a hemoglobin disorder, many types of cells contribute considerably to the pathophysiology of the disease. The adhesion of neutrophils to activated endothelium is critical in the pathophysiology of sickle cell disease and targeting neutrophils and their interactions with endothelium represents an important opportunity for the development of new therapeutics. We focused on endothelin-1, a mediator involved in neutrophil activation and recruitment in tissues, and investigated the involvement of the endothelin receptors in the interaction of neutrophils with endothelial cells. We used fluorescence intravital microscopy analyses of the microcirculation in sickle mice and quantitative microfluidic fluorescence microscopy of human blood. Both experiments on the mouse model and patients indicate that blocking endothelin receptors, particularly ETB receptor, strongly influences neutrophil recruitment under inflammatory conditions in sickle cell disease. We show that human neutrophils have functional ETB receptors with calcium signaling capability, leading to increased adhesion to the endothelium through effects on both endothelial cells and neutrophils. Intact ETB function was found to be required for tumor necrosis factor α-dependent upregulation of CD11b on neutrophils. Furthermore, we confirmed that human neutrophils synthesize endothelin-1, which may be involved in autocrine and paracrine pathophysiological actions. Thus, the endothelin-ETB axis should be considered as a cytokine-like potent pro-inflammatory pathway in sickle cell disease. Blockade of endothelin receptors, including ETB, may provide major benefits for preventing or treating vaso-occlusive crises in sickle cell patients.


Experiments with the site frequency spectrum.
Evaluating the likelihood function of parameters in highly-structured population genetic models from extant deoxyribonucleic acid (DNA) sequences is computationally prohibitive. In such cases, one may approximately infer the parameters from summary statistics of the data such as the site-frequency-spectrum (SFS) or its linear combinations. Such methods are known as approximate likelihood or Bayesian computations. Using a controlled lumped Markov chain and computational commutative algebraic methods, we compute the exact likelihood of the SFS and many classical linear combinations of it at a non-recombining locus that is neutrally evolving under the infinitely-many-sites mutation model. Using a partially ordered graph of coalescent experiments around the SFS, we provide a decision-theoretic framework for approximate sufficiency. We also extend a family of classical hypothesis tests of standard neutrality at a non-recombining locus based on the SFS to a more powerful version that conditions on the topological information provided by the SFS.