Relationship between Physical Activity, Screen Time and Weight Status among Young Adolescents.
It is well established that lack of physical activity and high bouts of sedentary behaviour are now associated with all-cause and cardiovascular mortality. The purpose of this study was to investigate the relationship between physical activity participation, overall screen time and weight status amongst early Irish adolescent youth. Participants were a sample of 169 students: 113 boys (mean age = 12.89 ± 0.34 years) and 56 girls (mean age = 12.87 ± 0.61 years). The data gathered in the present study included physical activity (accelerometry), screen time (self-report) and anthropometric measurements. Overweight and obese participants accumulated significantly more minutes of overall screen time daily compared to their normal-weight counterparts. A correlation between physical activity and daily television viewing was evident among girls. No significant interaction was apparent when examining daily physical activity and overall screen time in the prediction of early adolescents' body mass index. Results suggest the importance of reducing screen time in the contribution towards a healthier weight status among adolescents. Furthermore, physical activity appears largely unrelated to overall screen time in predicting adolescent weight status, suggesting that these variables may be independent markers of health in youth. The existing relationship for girls between moderate-to-vigorous physical activity and time spent television viewing may be a potential area to consider for future intervention design with adolescent youth.


Local structural properties of 0.5BiMnO3-0.5ATiO3 (A = Ba or Sr).
Neutron total scattering data have been used to probe the long- and short-range structure of 0.5BiMnO(3)-0.5ATiO(3) (A = Ba or Sr). Modelling of the total scattering data, using the reverse Monte Carlo (RMC) method, reveals local disorder that differs substantially from the average structure. Significantly, the Bi(3+) cations prefer to adopt an asymmetric coordination due to the influence of the lone-pair electrons.


Association of fresh vegetable and salt-preserved vegetable consumptions with estimated glomerular filtration rate.
This study aimed to investigate the relationship between the consumption of fresh and salt-preserved vegetables and the estimated glomerular filtration rate (eGFR), which requires further research. For this purpose, the data of those subjects who participated in the 2011-2012 and 2014 surveys of the Chinese Longitudinal Healthy Longevity Survey (CLHLS) and had biomarker data were selected. Fresh and salt-preserved vegetable consumptions were assessed at each wave. eGFR was assessed using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation based on plasma creatinine. Furthermore, a linear mixed model was used to evaluate associations between fresh/salt-preserved vegetables and eGFR. The results indicated that the median baseline and follow-up eGFRs were 72.47 mL/min/1.73 m² and 70.26 mL/min/1.73 m², respectively. After applying adjusted linear mixed model analysis to the data, the results revealed that compared to almost daily intake, occasional consumption of fresh vegetables was associated with a lower eGFR (β=-2.23, 95% CI: -4.23, -0.23). Moreover, rare or no consumption of salt-preserved vegetables was associated with a higher eGFR (β = 1.87, 95% CI: 0.12, 3.63) compared to individuals who consumed salt-preserved vegetables daily. Fresh vegetable consumption was direct, whereas intake of salt-preserved vegetables was inversely associated with eGFR among the oldest subjects, supporting the potential benefits of diet-rich fresh vegetables for improving eGFR.


Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange.
The development of programmable nucleases has enabled the application of new genome engineering strategies for cellular immunotherapy. While targeted nucleases have mostly been used to knock-out or knock-in genes in immune cells, the scarless exchange of entire immunogenomic alleles would be of great interest. In particular, reprogramming the polymorphic MHC locus could enable the creation of matched donors for allogeneic cellular transplantation. Here we show a proof-of-concept for reprogramming MHC-specificity by performing CRISPR-Cas9-assisted cassette exchange. Using murine antigen presenting cell lines (RAW264.7 macrophages), we demonstrate that the generation of Cas9-induced double-stranded breaks flanking the native MHC-I H2-Kd locus led to exchange of an orthogonal H2-Kb allele. MHC surface expression allowed for easy selection of reprogrammed cells by flow cytometry, thus obviating the need for additional selection markers. MHC-reprogrammed cells were fully functional as they could present H2-Kd-restricted peptide and activate cognate T cells. Finally, we investigated the role of various donor template formats on exchange efficiency, discovering that templates that underwent in situ linearization resulted in the highest MHC-reprogramming efficiency. These findings highlight a potential new approach for the correcting of MHC mismatches in cellular transplantation.


Hsp47 a novel collagen binding serpin chaperone, autoantigen and therapeutic target.
Hsp47 was originally discovered as a cell surface collagen binding protein, colligin, and was later shown to be an endoplasmic reticulum (ER) resident protein with collagen binding properties in chick fibroblasts. Hsp47 has been termed J6, gp46, CB48 and CBP2 in various other organisms and has been mapped to human chromosome 11q13.5 a known "hot spot" in a number of human cancers. Hsp47 has been shown to be constitutively expressed with collagens; it is heat inducible and binds to both helical and non-helical forms of collagens. Hsp47 binds closely to procollagen in the ER, but dissociates from it in the cis-Golgi to allow fibril formation. Hsp47 is over-expressed in many fibrotic diseases including: glomerulosclerosis, pulmonary fibrosis, liver cirrhosis, cicatricial pemphigoid, epidermolysis bullosa acquista and keloids. Hsp47 is associated with fibrosis following myocardial infarction and has been localized in artherosclerotic arteries. Among a number of rheumatoid conditions, Hsp47 manifests properties of an autoantigen and in some cancers appears to be a biomarker. The unique properties of Hsp47 in modulating collagen production and its location to the cell membrane in many forms of cancer have designated Hsp47 as a potential biomarker and/or therapeutic target for a number of conditions and diseases.


Visible Light Mediated Co-Catalyzed Isocyanide Insertion with Sulfonyl Azide: Synthesis of Sulfonyl Carbamimidic Azide and Sulfonyl Aminotetrazole via Carbodiimide Intermediate.
Herein, we report an operationally simple and efficient protocol to prepare sulfonyl carbamimidic azide and N-sulfonyl aminotetrazole via Co-catalyzed three component coupling of sulfonyl azide (acts as nitrene source), isocyanide, and TMS-azide at room temperature under visible light. Initially, the carbamimidic azide is formed, which cyclizes only in the presence of base to deliver N-sulfonyl aminotetrazole in very good yields. The sulfonyl aminotetrazole can also be synthesized directly without isolating the carbamimidic azide in the presence of base. The sulfonyl azide is anticipated to generate nitrene and reacts with isocyanide to produce carbodiimide. Subsequent addition of azide (TMS-N3 ) to carbodiimide results in the formation of carbamimidic azide.


The Effects of Race, Ethnicity, and Maternal Education on Infant Mortality.
The state of New Jersey has a large Black/African American (AA) versus White racial disparity in infant mortality and educational level at childbirth. This disparity, measured by rate ratio, increases with greater maternal education among varied racial-ethnic groups. The nature of this disparity measured by rate differences has not been explored. Infant birth and mortality data were used to examine whether racial or ethnic disparities in infant mortality increased with greater maternal education, comparing rate differences and rate ratios. Racial and ethnic variations in the association between maternal education and infant mortality were examined. Data were from the New Jersey State Health Assessment Data for all New Jersey births between 2014 and 2018 stratified by race and ethnicity, maternal education, and infant mortality ( n = 481,333). R software was used to create a data set and estimate additive and multiplicative interactions, rate differences, and rate ratios for infant mortality by maternal race/ethnicity and educational levels among four racial-ethnic groups. Infant mortality was significantly greater for Black/AA and Hispanic mothers than for White mothers. At all educational levels, Black/AA mothers had the highest prevalence of infant mortality compared to other racial or ethnic groups. Rate differences in infant mortality showed a decrease in Black/AA-White differences for mothers with a high school education or less compared to mothers with a college degree. However, rate ratios showed an increase in Black/AA-White ratio with increasing education levels for mothers with high school education or less than mothers with a college degree. Risk ratios comparing infant mortality for Black/AA versus Hispanic or Asian mothers showed more than a twofold greater risk at all education levels for Black/AA infants. Finally, college-educated Black/AA mothers had significantly higher rates of infant mortality than White or Hispanic mothers with a high school education or less. Black/AA mothers with a college degree had a higher infant mortality rate than White, Hispanic, or Asian mothers with a high school education or less. Future research should address contextual/systemic contributors to this disparity.


Epithelium-generated neuropeptide Y induces smooth muscle contraction to promote airway hyperresponsiveness.
Asthma is one of the most common chronic diseases globally and can be divided into presenting with or without an immune response. Current therapies have little effect on nonimmune disease, and the mechanisms that drive this type of asthma are poorly understood. Here, we have shown that loss of the transcription factors forkhead box P1 (Foxp1) and Foxp4, which are critical for lung epithelial development, in the adult airway epithelium evokes a non-Th2 asthma phenotype that is characterized by airway hyperresponsiveness (AHR) without eosinophilic inflammation. Transcriptome analysis revealed that loss of Foxp1 and Foxp4 expression induces ectopic expression of neuropeptide Y (Npy), which has been reported to be present in the airways of asthma patients, but whose importance in disease pathogenesis remains unclear. Treatment of human lung airway explants with recombinant NPY increased airway contractility. Conversely, loss of Npy in Foxp1- and Foxp4-mutant airway epithelium rescued the AHR phenotype. We determined that NPY promotes AHR through the induction of Rho kinase activity and phosphorylation of myosin light chain, which induces airway smooth muscle contraction. Together, these studies highlight the importance of paracrine signals from the airway epithelium to the underlying smooth muscle to induce AHR and suggest that therapies targeting epithelial induction of this phenotype may prove useful in treatment of noneosinophilic asthma.


First evidence of melatonin receptors distribution in the suprachiasmatic nucleus of tree shrew brain.
In this study we investigated the distribution of melatonergic receptors in the tree shrew brain. The psychosocial stress in the tree shrew is a validated model of depression with disturbance of circadian rhythms. Given the role of melatonin in the modulation of circadian rhythms, we determined by autoradiography the distribution of binding sites of 2-[125I]-MLT, a ligand for the two melatonergic receptors MT1 and MT2, in the tree shrew brain focusing mostly on the suprachiasmatic nucleus (SCN), the biological clock involved in resynchronization of circadian rhythms. We also analyzed the distribution of 2-[125I]-MLT in other brain areas involved in regulation of mood. Specific binding of 2-[125I]-MLT was found in the SCN. In addition, several structures in the tree shrew brain were labeled, among them the pars tuberalis, the cerebellum, structures of the hippocampal formation and dopaminergic areas such as the caudate putamen and nucleus accumbens, providing other potential targets of psychosocial stress in the tree shrew, in addition to the SCN. The demonstration of melatonergic receptors in these brain areas supports their probable involvement in the behavioural, neuroendocrine and circadian rhythms disturbances observed in the psychosocial stress model of depression in the tree shrew.


Physiology and biochemistry of endothelial function in children with chronic renal failure.
Premature atherosclerosis is a major cause of morbidity and mortality in chronic renal failure (CRF). Endothelial dysfunction is a key early event in atherogenesis. The aim of this study was to assess the effect of CRF on endothelial function using physiological and biochemical measures. To focus on the effect of CRF itself, 23 children (matched with 23 controls for age and vessel diameter) were selected because they were normotensive, had normal total cholesterol (TC) levels, and were not on vasoactive drugs. Their mean (range) age was 12.0 (7.8 to 17.0) years; GFR 17.5 (8.8 to 34.5) ml/min/1.73 m2. The physiology of endothelial function in the brachial artery was assessed using high resolution ultrasound by measuring its diameter at rest, during reactive hyperemia (endothelium dependent dilation) and after sublingual glyceryl trinitrate (GTN; endothelium independent dilation). Nitric oxide (NO) metabolites and endogenous NO synthetase (eNOS) inhibitors were measured as an assessment of endothelial metabolism. Brachial artery dilation to flow [FMD, mean (SEM)%] was reduced in CRF to 4.9 (0.6) and controls 8.6 (0.6), P < 0.0001. In contrast, the response to GTN was similar in both groups: CRF 25.1 (1.6), controls 23.3 (1.2), P = 0.31. There was no difference in TC, low density lipoprotein (LDL) or high density lipoprotein (HDL) between the patients and the controls. Triglycerides (TG) were higher in the patients but within the normal range. Antibodies against oxidized LDL (ox-LDL) were high in CRF. Endogenous NOS inhibitors were high in CRF, and intermediate NO metabolites were low. There was no correlation between FMD of the brachial artery and lipid subfractions, or with NO metabolites or eNOS inhibitors. Endothelium dependent dilation of the brachial artery is impaired in children with CRF who do not have co-existing risk factors for atherosclerosis. This may represent early evidence of atherogenic vascular disease.


Clinical use of genotypic and phenotypic drug resistance testing to monitor antiretroviral chemotherapy.
Assays that detect antiretroviral drug resistance in human immunodeficiency virus have recently become available to clinicians. Phenotypic assays measure the drug susceptibility of the virus by determining the concentration of drug that inhibits viral replication in tissue culture. Genotypic assays determine the presence of mutations that are known to confer decreased drug susceptibility. Although each type of assay has specific advantages, limitations associated with these tests often complicate the interpretation of results. Several retrospective clinical trials have suggested that resistance testing may be useful in the assessment of the success of salvage antiretroviral therapy. Prospective, controlled trials have demonstrated that resistance testing improves short-term virological response. Resistance testing is currently recommended to help guide the choice of new drugs for patients after treatment has failed and for pregnant women. Resistance testing should also be considered for treatment-naïve patients, to detect transmission of resistant virus.


Functionalized Magnetic PLGA Nanospheres for Targeting and Bioimaging of Breast Cancer.
Superparamagnetic iron oxide nanoparticles (SPIONs) are actively used as highly sensitive imaging probes to provide contrast in MRI. In this study, we propose the use of SPIONs encapsulated with antibody-conjugated poly(lactic-co-glycolic acid) (PLGA) as a potent theragnostic agent. The SPIONs were synthesized by a chemical co-precipitation method of ferric and ferrous ions, and subsequently encapsulated with PLGA by using an emulsification-diffusion method. Herceptin was chemically conjugated to the SPION-encapsulating PLGA nanoparticles to target the human epidermal growth factor receptor 2 (Her2/neu) overexpressing breast cancers. FACS and MR molecular imaging revealed that the Her2/neu overexpressing cell line showed a stronger contrast enhancement than the Her2/neu non-expressing cell lines, and the signal intensity of in vivo MR imaging decreased as the concentration of Herceptin increased. This strategy of encapsulating SPIONs with PLGA will be highly useful in functionalizing magnetic nanoparticles and improving the diagnostic and therapeutic efficacy of a wide array of cancer treatments.


Expression of the interleukin-4 receptor alpha in human conjunctival epithelial cells.
To investigate the expression and function of interleukin-4 receptor alpha (IL-4Ralpha) in human conjunctival epithelial cells (HCjECs). The presence of IL-4Ralpha mRNA and protein was examined by reverse transcription (RT) PCR and immunohistology, respectively. Cell surface expression was examined by flow cytometry. The effects of interleukin (IL)-4 or IL-13 on the tyrosine phosphorylation of signal transducer and the activator of transcription 6 (STAT6) were evaluated by immunoblot analyses. The transcripts upregulated upon IL-4 stimulation were examined using GeneChip, and confirmed by quantitative RT-PCR. IL-4Ralpha mRNA and protein were detected in human conjunctival epithelium. IL-4Ralpha protein was expressed on the cell surface of HCjECs. IL-4 and IL-13 induced tyrosine phosphorylation of STAT6. GeneChip analysis showed that nine transcripts were upregulated more than fourfold by IL-4 stimulation in the primary HCjECs from two individuals. Quantitative RT-PCR assay confirmed the upregulation of these transcripts: lecithin retinol acyltransferase (LRAT), calpain (CAPN14), tumour necrosis factor alpha-induced protein 6 (TNFAIP6), RAS guanyl-releasing protein 1 (RASGRP1), endothelin receptor type A (EDNRA), hyaluronan synthase 3 (HAS3), cathepsin C (CTSC), carbonic anhydrase II (CA2) and cytokine-inducible SH2-containing protein (CISH). HCjECs expressed functioning IL-4Ralpha, and IL-4 stimulation induced the expression of several genes.


MSCohi-O lenses for long-term retention of mesenchymal stem cells on ocular surface as a therapeutic approach for chronic ocular graft-versus-host disease.
Chronic ocular graft-versus-host disease (oGVHD) is a common complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and can lead to vision loss if not diagnosed and treated promptly. Currently, no approved drugs exist for oGVHD treatment. However, umbilical cord-derived mesenchymal stem cells (UCMSCs) have known immunoregulatory properties and have been employed in clinical trials for immune-mediated diseases. To address oGVHD, the application of UCMSCs to the ocular surface is a logical approach. Intravenous administration of UCMSCs poses risks, necessitating topical and local delivery. Retaining UCMSCs on the ocular surface remains a challenge. To overcome this, we invented mesenchymal stem cell-coating high oxygen-permeable hydrogel lenses combining UCMSCs and machinery to enable the long-term retention of UCMSCs on the ocular surface. Animal model experiments demonstrated that these lenses effectively retained UCMSCs, providing therapeutic benefits by decreasing corneal inflammation and damage, and inhibiting immune rejection and response, all crucial aspects in oGVHD treatment.


(E)-13-(4-Amino-phen-yl)parthenolide.
The title compound, C21H25NO3 [systematic name: (3aS,9aR,10aR,10bS,E)-3-[(E)-4-(4-amino-benzyl-idene)-6,9a-dimethyl-3a,4,5,8,9,9a,10a,10b-octa-hydro-oxireno[2',3':9,10]cyclo-deca-[1,2-b]furan-2(3H)-one] was obtained from the reaction of parthenolide [synonym: 4,5-ep-oxy-germacra-1(10),11(13)-dieno-12,6-lactone] with 4-iodo-aniline under Heck reaction conditions. It was identified as the E-isomer (conformation about the exocyclic methyl-idene C=C bond; the conformation about the C=C bond in the ten-membered ring is also E). The mol-ecule is built up from fused ten-, five- (lactone) and three-membered (epoxide) rings with a 4-amino-phenyl group as a substituent. The ten-membered ring displays an approximate chair-chair conformation, while the lactone ring has an envelope conformation with the C atom bonded to the ring O atom as the flap. The dihedral angle between the benzene ring of the 4-amino-phenyl moiety and the lactone ring mean plane is 23.50 (8)°. In the crystal, mol-ecules are linked via N-H⋯O hydrogen bonds, between the amine group and the lactone and epoxide ring O atoms, forming chains propagating along the b-axis direction. Adjacent chains are linked via C-H⋯O inter-actions, forming an undulating two-dimensional network lying parallel to the plane (001). The absolute structure of the mol-ecule in the crystal was confirmed by resonance scattering [Flack parameter = 0.03 (3)].


Merino sheep: a further look at quantitative trait loci for wool production.
A quantitative trait loci (QTL) analysis of wool traits from experimental half-sib data of Merino sheep is presented. A total of 617 animals distributed in 10 families were genotyped for 36 microsatellite markers on four ovine chromosomes OAR1, OAR3, OAR4 and OAR11. The markers covering OAR3 and OAR11 were densely spaced, at an average distance of 2.8 and 1.2 cM, respectively. Body weight and wool traits were measured at first and second shearing. Analyses were conducted under three hypotheses: (i) a single QTL controlling a single trait (for multimarker regression models); (ii) two linked QTLs controlling a single trait (using maximum likelihood techniques) and (iii) a single QTL controlling more than one trait (also using maximum likelihood techniques). One QTL was identified for several wool traits on OAR1 (average curvature of fibre at first and second shearing, and clean wool yield measured at second shearing) and on OAR11 (weight and staple strength at first shearing, and coefficient of variation of fibre diameter at second shearing). In addition, one QTL was detected on OAR4 affecting weight measured at second shearing. The results of the single trait method and the two-QTL hypotheses showed an additional QTL segregating on OAR11 (for greasy fleece weight at first shearing and clean wool yield trait at second shearing). Pleiotropic QTLs (controlling more than one trait) were found on OAR1 (clean wool yield, average curvature of fibre, clean and greasy fleece weightand staple length, all measured at second shearing).


Effect of pentylentetrazol-induced seizures on A1 adenosine receptor regional density in the mouse brain: a quantitative autoradiographic study.
Adenosine has been shown to be a major regulator of neuronal activity in convulsive disorders, exerting its anticonvulsant effect through central A1 adenosine receptors. The aim of the present study was to investigate the effect of generalized tonic-clonic seizures induced by pentylentetrazol on regional changes in A1 adenosine receptor density and distribution in the mouse brain by in vitro quantitative autoradiography. As radioligand the specific agonist of A1 receptors [3H]cyclohexyladenosine was used. After two consecutive (once daily) pentylentetrazol-induced convulsions a widespread upregulation of A1 receptor density was detected with a marked enhancement in structures that mediate seizure activity like hippocampus, mamillary bodies, septum, substantia nigra, thalamic nuclei and cerebral cortices. On the contrary, in basal ganglia a significant downregulation of A1 receptors was observed. These results indicate that: (i) the observed increases or decreases in A1 receptor density are organized in selective anatomical structures related to seizure development rather than uniform in the brain; and (ii) since the upregulation of A1 receptors is sufficient to enhance the physiological depressive response of adenosine, the overall evoked increases seen here may lead to a stronger inhibitory tone and accordingly to a more efficient anticonvulsant effect of endogenous adenosine.


Apoptotic pathways of macrophages within osteolytic interface membrane in periprosthestic osteolysis after total hip replacement.
Macrophage apoptosis in interface membrane, which occurs through either death receptor, mitochondrion, or endoplasmic reticulum (ER) stress pathways, has been suggested to play an important role in promoting osteolysis. However, how and why macrophage apoptosis originates and the correlation among these apoptotic pathways is not yet clear. The objective of this study was to identify the apoptotic mechanism of macrophages, and to explore the relationship between the apoptotic pathways and progression of osteolysis. Transmission electron microscopy (TEM) was utilized to analyze the tissue ultrastructure of wear particles, and in situ apoptotic macrophage identification was performed by TUNEL staining. We analyzed the expression of the key biomarkers of apoptotic pathways via immunohistochemistry and Western blotting. Our results demonstrated that the majority of wear particles within osteolytic interface membrane was in the 30-60 nm range, and that macrophage apoptotic ratio increased along with osteolysis progression. Normal hip dysplasia and mechanical loosening of tissues showed low expression levels of biomarkers for ER stress (Ca2+ , JNK, cleaved Caspase-4, IRE1-α, Grp78/Bip, and CHOP), mitochondrion (Bcl-2, Bax, and Cytochrome c), and death receptor (Fas and cleaved Caspase-8) pathways, while osteolytic interface membrane tissues expressed high levels of these biomarkers. In addition, we found that the ER stress intensity was in complete conformity with mitochondrial dysfunction and was consistent with the results of death receptor activation. Thus, our findings suggested that wear particles generated at implant interface can accelerate macrophage apoptosis through changes in apoptotic pathways and ultimately aggravate the symptom of osteolysis. These data represent a preferential apoptotic signaling pathway of macrophages as specific target points for the prevention and therapeutic modulation of periprosthetic osteolysis.


Metabolic syndrome in polycystic ovary syndrome.
Because insulin resistance and visceral obesity are important features of polycystic ovary syndrome (PCOS), metabolic syndrome is much more common in women with PCOS than in the general female population of similar age. It has been reported that in the USA almost 50% of women with PCOS present the metabolic syndrome. In Italy, where women with PCOS have a lower mean body weight and less frequently increased serum triglycerides than US PCOS, metabolic syndrome is less common but still 4 times more frequent in PCOS patients than in the general female population of similar age. Patients with mild PCOS phenotype (ovulatory PCOS) have a lower prevalence of metabolic syndrome but, in these patients too, metabolic syndrome is 2 times more frequent than in the normal population. These data suggest that PCOS is the most common cause of increased cardiovascular risk in young adult women. All obese and overweight women with PCOS should be screened for metabolic syndrome and, when the syndrome is not found, the screening should be repeated every 2 or 3 years. Treatment consists in lifestyle intervention. Pharmacological therapies should be used only when lifestyle fails to normalize cardiovascular risk factors.


Long-term Changes in Grassland Soil Phosphorus with Fertilizer Application and Withdrawal.
Long-term phosphorus (P) applications can increase soil P concentrations in excess of agronomic optima, posing a risk to water quality. Once fertilization stops, however, it may take time for soil P concentrations to decline. Whereas P fertilization adds orthophosphate, little is known about changes in other soil P forms during P buildup and drawdown. This study examined changes in P pools (total P, Olsen P, Mehlich P, and water-extractable P) and P forms determined by P-nuclear magnetic resonance spectroscopy (P-NMR) in grazed grassland plots from Northern Ireland. Between 1994 and 1999, all plots received 8.3 kg P ha yr with variable rates of nitrogen (100-500 kg N ha yr). From 2000 to 2005, plots received 0, 20, 40, or 80 kg P ha yr and 250 kg N ha yr; from 2005 to 2010, no P fertilizer was applied to any plots. In 2005, soil P pool concentrations at the highest P fertilization rates were significantly elevated compared with those in 2000 but had decreased to 2000 concentrations by 2010. In soils receiving no P, soil P pool concentrations were significantly lower than those in 1994 only in 2010. There were few changes in P forms determined by P-NMR. Orthophosphate followed the same trend observed for the soil P pools; total organic P, total inositol phosphates, and total orthophosphate monoesters and diesters were highest in 2010 in the soil receiving no P fertilizer for 10 yr. For these soils, fertilizer application and cessation influenced inorganic P more than organic P.


A Health Care Systems Approach to Improving Care for Seriously Ill Patients.
Health care systems can go beyond advance care planning to create mechanisms for eliciting and documenting the goals of care and life-sustaining treatment decisions of patients with serious life-limiting illnesses. These systems can help ensure that patients receive care that is consistent with their values and preferences. We describe a case in which even though a patient with a serious illness had completed an advance directive and had discussed preferences with family, clinicians failed to identify the patient's authentic preferences for life-sustaining treatment. We offer a stepwise framework for communication with seriously ill patients and describe a systems approach to transforming the process of eliciting, documenting, and honoring patients' life-sustaining treatment preferences in the U. S. Veterans Health Administration.


Perceived ethnic discrimination versus acculturation stress: influences on substance use among Latino youth in the Southwest.
Using a predominately Mexican-origin Latino sample of 5th grade students from the Southwestern United States, this study examined the relative effects of perceived discrimination and acculturation stress on substance use, and it assessed whether these effects were moderated by linguistic acculturation or time in the United States. Although rates of substance use were generally low in the sample, given the young age of the participants, over half (59%) of the sample perceived some discrimination, and almost half (47%) experienced some acculturation stress. Spanish-dominant and bilingual youth perceived more discrimination than English-dominant youth, whereas youth who have been in the United States five or fewer years perceived more discrimination than youth with more time in the United States. Youth who were Spanish-dominant or were recent arrivals experienced the most acculturation stress, with levels declining as linguistic acculturation and time in the United States increased. Multiple regression estimates indicated that perceived discrimination was associated with larger amounts and higher frequency of recent substance use and an array of substance use attitudes, such as stronger intentions to use substances, espousal of pro-drug norms, more positive substance use expectancies, and peer approval of substance use. Although acculturation stress was not associated with substance use, it was positively associated with several substance use attitudes, which are known antecedents of actual use. With a few exceptions, linguistic acculturation and time in the United States did not moderate the effects of perceived discrimination or acculturation stress.


Parvovirus B19 infection in a human immunodeficiency virus-infected patient with anemia.
Anemia is generally attributed to zidovudine therapy in human immunodeficiency virus (HIV)-infected patients, although parvovirus B19 infection has been reported as a rare cause. We report on a 24-year-old homosexual man infected with HIV who presented with anemia. He had received aggressive daily antiretroviral therapy (zidovudine 600 mg, lamivudine 300 mg, and saquinavir 1,800 mg) for 2 years. At the time of admission, his CD4+ count was 10 x 10(6) cells/L. A bone marrow aspirate smear showed a marked decrease in erythropoiesis and immunocytochemical staining for parvovirus B19 was positive. Parvovirus B19 viral DNA was detected in the peripheral blood using a polymerase chain reaction-based assay. Serologic studies were positive for parvovirus B19 immunoglobulin (Ig)M antibodies, but negative for IgG antibodies. The patient was treated with packed red blood cell transfusion. Zidovudine was stopped and replaced with zalcitibine 2.25 mg daily after anemia occurred. He did not receive intravenous Ig therapy because of its cost. After discontinuation of zidovudine for 1 year, anemia persisted and the patient depended on regular blood transfusions to control the anemia. This case emphasizes that, in addition to drug-related causes, parvovirus B19 infection should be included in the differential diagnosis of chronic anemia in HIV-infected individuals.


A Quantitative Theory for Genomic Offset Statistics.
Genomic offset statistics predict the maladaptation of populations to rapid habitat alteration based on association of genotypes with environmental variation. Despite substantial evidence for empirical validity, genomic offset statistics have well-identified limitations, and lack a theory that would facilitate interpretations of predicted values. Here, we clarified the theoretical relationships between genomic offset statistics and unobserved fitness traits controlled by environmentally selected loci and proposed a geometric measure to predict fitness after rapid change in local environment. The predictions of our theory were verified in computer simulations and in empirical data on African pearl millet (Cenchrus americanus) obtained from a common garden experiment. Our results proposed a unified perspective on genomic offset statistics and provided a theoretical foundation necessary when considering their potential application in conservation management in the face of environmental change.


Dementia Caregiving Outcomes: The Impact of Caregiving Onset, Cognitive Impairment and Behavioral Problems.
With a projected increase in the prevalence of Alzheimer's disease (AD) and related dementias, family caregiving is steadily increasing. Caring for an individual with AD is characterized as a "career," and within this career are a number of key transitions, including the onset of caregiving. Research reveals a number of negative emotional health outcomes for AD caregivers, including depression and role overload, but less research has examined the influence of key transitions on the emotional adaptation of caregivers. The purpose of this study is to examine how different patterns of caregiving onset (gradual and abrupt) impact emotional health outcomes for AD caregivers. This study also explores how care-recipient cognitive impairment and behavioral problems moderate these relationships. Cross-sectional, quantitative data from 100 caregivers completing self-administered surveys were used. A series of one-way ANOVAs and multiple regression analyses were conducted to address study aims. Results indicate that care-recipient cognition and behavioral problems moderated the relationship between caregiving onset and emotional health outcomes for caregivers who experienced a more abrupt entry into the caregiving role. Results suggest the importance of considering different onset transitions and moderating factors within the caregiving career to target clinical interventions. Implications for social work practice are discussed.


The influence of club-head kinematics on early ball flight characteristics in the golf drive.
Despite many coaching and biomechanical texts describing how the kinematics of the club-head at impact lead to distance and accuracy of the ball flight, there is limited quantitative evidence supporting these assertions. The purpose of this study was to quantify the relationships between club-head kinematics and subsequent early ball flight characteristics during the golf drive. An opto-reflective system operating at 400 Hz was used to capture the swings of 21 male golfers using their own drivers. The 3D displacement data permitted the calculation of club-head kinematics at impact, as well as subsequent early ball flight characteristics. Using regression analyses, club-head kinematics at impact (velocity, orientation, path, and centeredness) were used to explain the variability in five dependent variables of early ball flight characteristics (resultant velocity, launch angle, side angle, back spin, and side spin). The results of the study indicated that club-head kinematics at impact explained a significant proportion of early ball flight characteristics (adjusted r2 = 0.71-0.82), even when generalized across individual clubs.


Current concepts: laser treatment of adult vascular lesions.
Patients frequently seek treatment of cutaneous vascular lesions for both medical and cosmetic reasons. Advances in the use of lasers and light sources enable physicians to effectively treat vascular lesions that were previously untreatable. Given the plethora of devices available, selecting the appropriate laser system and treatment parameters can be confusing for the laser surgeon. This article reviews current concepts in the treatment of adult vascular conditions. It highlights the different types of lasers and the most common vascular lesions amenable to therapy. Also, the article offers several pearls regarding selection of the laser and of the treatment parameters.


Extensive changes in DNA methylation are associated with expression of mutant huntingtin.
The earliest stages of Huntington disease are marked by changes in gene expression that are caused in an indirect and poorly understood manner by polyglutamine expansions in the huntingtin (HTT) protein. To explore the hypothesis that DNA methylation may be altered in cells expressing mutated HTT, we use reduced representation bisulfite sequencing (RRBS) to map sites of DNA methylation in cells carrying either wild-type or mutant HTT. We find that a large fraction of the genes that change in expression in the presence of mutant huntingtin demonstrate significant changes in DNA methylation. Regions with low CpG content, which have previously been shown to undergo methylation changes in response to neuronal activity, are disproportionately affected. On the basis of the sequence of regions that change in methylation, we identify AP-1 and SOX2 as transcriptional regulators associated with DNA methylation changes, and we confirm these hypotheses using genome-wide chromatin immunoprecipitation sequencing (ChIP-Seq). Our findings suggest new mechanisms for the effects of polyglutamine-expanded HTT. These results also raise important questions about the potential effects of changes in DNA methylation on neurogenesis and cognitive decline in patients with Huntington disease.


Single microassay for matrix degrading enzymes.
Matrix degrading enzymes are implicated in several disease processes such as abdominal aortic aneurysms and emphysema, however, monitoring proteolytic activity in a single assay is not well-established. Numerous assays have been developed to measure matrix degrading enzymes, which use artificial substrates or substrates derived from natural substrate protein. We have recently developed an assay for elastolytic activity based on the detection of primary amines, using trinitrobenzene sulfonic acid (TNBSA), following the digestion of succinylated elastin. The assay is also versatile enough to allow the detection of other proteases through the use of succinylated substrate specific for given protease. In order to improve the sensitivity and versatility of the assay we have refined the buffer conditions (PBS pH 7.2/1 mM CaCl2) to provide a 60 % increase in sensitivity to elastolytic activity and also formulated a substrate mixture containing succinylated elastin, collagen and gelatin. The combination of a substrate mixture and an optimum buffer will allow a spectrum of enzymes to be detected in a single reaction, providing a more complete picture of total proteolytic activity in a biological sample. This assay may also provide a tool to use proteolytic activity as a marker to monitor pathologic conditions involving matrix turn-over.


Comparison of virtual reality exergaming and home exercise programs in patients with subacromial impingement syndrome and scapular dyskinesis: Short term effect.
The aim of this study was to compare the short term effects of home exercise program and virtual reality exergaming in patients with subacromial impingement syndrome (SAIS). A total of 30 patients with SAIS were randomized into two groups which are Home Exercise Program (EX Group) (mean age: 40.6 ± 11.7 years) and Virtual Reality Exergaming Program (WII Group) (mean age: 40.33 ± 13.2 years). Subjects were assessed at the first session, at the end of the treatment (6 weeks) and at 1 month follow-up. The groups were assessed and compared with Visual Analogue Scale (based on rest, activity and night pain), Neer and Hawkins Tests, Scapular Retraction Test (SRT), Scapular Assistance Test (SAT), Lateral Scapular Slide Test (LSST) and shoulder disability (Shoulder Pain and Disability Index (SPADI)). Intensity of pain was significantly decreased in both groups with the treatment (p < 0.05). The WII Group had significantly better results for all Neer test, SRT and SAT than the EX Group (p < 0.05). Virtual reality exergaming programs with these programs were found more effective than home exercise programs at short term in subjects with SAIS. Level I, Therapeutic study.


IL-12 gene therapy is an effective therapeutic strategy for hepatocellular carcinoma in immunosuppressed mice.
Immunosuppressive therapy for organ transplantation is essential for controlling rejection. When liver transplantation is performed as a therapy for hepatocellular carcinoma (HCC), recurrent HCC is one of the most fatal complications. In this study, we show that intratumoral murine IL-12 (mIL-12) gene therapy has the potential to be an effective treatment for malignancies under immunosuppression. C3H mice (H-2(k)), injected with FK506 (3 mg/kg) i.p., were s.c. implanted with 2.5 x 10(6) MH134 cells (H-2(k)) and we treated the established HCC with electroporation-mediated gene therapy using mIL-12 plasmid DNA. Intratumoral gene transfer of mIL-12 elevated intratumoral mIL-12, IFN-gamma, and IFN-gamma-inducible protein-10, significantly reduced the number of microvessels and inhibited the growth of HCC, compared with HCC-transferred control pCAGGS plasmid. The inhibition of tumor growth in immunosuppressed mice was comparable with that of mIL-12 gene therapy in immunocompetent mice. Intratumoral mIL-12 gene therapy enhanced lymphocytic infiltration into the tumor and elicited the MH134-specific CTL response even under FK506. The dose of FK506 was sufficient to prevent the rejection of distant allogenic skin grafts (BALB/c mice, H-2(d)) and tumors, B7-p815 (H-2(d)) used as transplants, during mIL-12 gene therapy against MH134. Ab-mediated depletion studies suggested that the inhibition of tumor growth, neovascularization, and spontaneous lung metastasis by mIL-12 was dependent almost entirely on NK cells and partially on T cells. These results suggest that intratumoral mIL-12 gene therapy is a potent effective strategy not only to treat recurrences of HCC in liver transplantation, but also to treat solid malignant tumors in immunosuppressed patients with transplanted organ.


Identification of gamma-aminobutyric acid and its binding sites in Caenorhabditis elegans.
Gamma-aminobutyric acid (GABA), glutamate decarboxylase and GABA-transaminase were identified in the nematode Caenorhabditis elegans. The concentration of GABA in C. elegans (0.14 micrograms/mg protein) is approximately 10-fold lower than the concentration of GABA in rat brain. Glutamate decarboxylase and GABA-transaminase, the GABA anabolic and catabolic enzymes, are also present in C. elegans. Crude membrane fractions were prepared from C. elegans and used to study specific [3H] GABA binding sites. GABA binds to C. elegans membranes with high affinity (37 nM) and low capacity (Bmax = 2.25 pmol/mg protein). Muscimol is a competitive inhibitor of specific GABA binding with a KI value of 120 nM. None of the other GABA agonists or antagonists inhibited greater than 40% of the specific GABA binding at concentrations up to 10(-4)M. Thirteen spider venoms were examined as possible GABA agonists or antagonists, the venom from Calilena agelenidae inhibits specific GABA binding with a KI value of 6 nl/ml. These results suggest that GABA has a physiological role as a neurotransmitter in C. elegans.


Cluster randomized trial of an antibiotic time-out led by a team-based pharmacist.
Antibiotic time-outs (ATOs) have been advocated to improve antibiotic use without dedicated stewardship resources, but their utility is poorly defined. We sought to evaluate the effectiveness of an ATO led by a team-based pharmacist. Cluster randomized controlled trial. Six medicine teams at an academic medical facility. Inpatients who received antibiotics and were cared for by a medicine team. In phase A (2 months) pharmacist-led ATOs were implemented on 3 medicine teams (ATO-A) while 3 teams maintained usual care (UC-A). In phase B (2 months), ATOs were continued in the ATO group (ATO-B) and ATOs were initiated in the UC group (UC ATO-B). We targeted 2 ATO points: early (<72 hours after antibiotics were initiated) and late (after the early period but ≤5 days after antibiotic initiation). In total, 290 ATOs were documented (181 early, 87 late, and 22 subsequent) among 538 admissions. The most common ATO recommendations were narrow therapy (148 of 290), no change (124 of 290), and change to oral (30 of 290). ATO initiation was lower in the UC ATO-B group than in either ATO group (21.8% UC ATO-B vs 69.2% ATO-A and -B). Overall antibiotic use was not different between the groups (P = .51), although intravenous (IV) levofloxacin use decreased in the UC group after ATO implementation (49 DOT/1,000 PD vs 20 DOT/1,000 PD; P = .022). The ratio of oral (PO) to intravenous (IV) DOT was lower in the UC group than in any of the ATO groups (P = .032). We detected no differences in mortality, length of stay, readmission, C. difficile infection, or antibiotic adverse events. Implementation of a pharmacist-led ATO was feasible and well accepted but did not change overall antibiotic use. An ATO may promote increased use of oral antibiotics, but more effective strategies for self-stewardship are needed.


Naltrexone administration affects ad libitum smoking behavior.
Endogenous opioid peptides have been implicated in the reinforcement of smoking and opioid antagonists have been examined to determine their role in smoking behavior. To date, the relationship between smoking behavior and chronic opiate antagonist administration during ad libitum smoking has not been investigated. The purpose of this study was to examine the relationships between naltrexone, an opiate antagonist administered orally, and smoking behavior and mood states during ad libitum smoking. A repeated measures experimental design was used. Normal adult male and female volunteers, admitted to the Clinical Research Center, were randomly assigned to naltrexone-treated (n = 22) or placebo-control (n = 21) groups in a double-blind manner. Day 1 was considered acclimation to the unit and day 2 was baseline, or pre-drug administration. On days 3, 4, and 5, subjects received 50 mg naltrexone or a placebo at 0700 and 1600 hours. Plasma nicotine and expired air carbon monoxide levels were measured daily at 1900 hours. Number of cigarettes smoked, mood states, withdrawal symptomatology and self-reported satisfaction with smoking were also quantified daily. Results indicated that plasma nicotine levels (P = 0.005), number of cigarettes smoked daily (P = 0.003) and self-reported satisfaction with smoking (P = 0.043) were significantly lower among those treated with naltrexone, compared to the placebo-control group. Expired air carbon monoxide levels did not differ between the two groups. In addition, mood states and withdrawal symptoms did not differ between groups. These findings suggest that endogenous opioid peptides influence specific smoking behavior variables.


A beta-fluoroamine inhibitor of purine nucleoside phosphorylase.
The potent immucillin purine nucleoside phosphorylase (PNP ) inhibitors F-DADMe-ImmH [(3S,4S)-3], and [(3R,4R)-3] are synthesized in seven steps. Cycloaddition to a fluoroalkene and an enzymic resolution are the key features of the construction of the fluoropyrrolidines 11, from which the immucillins are assembled by use of a three-component Mannich reaction. Slow-onset binding constants (Ki(*)) for [(3S,4S)-3] and [(3R,4R)-3] with human PNP are 0.032 and 1.82 nM, respectively. F-DADMe-ImmH [(3S,4S)-3] exhibits oral availability in mice at doses as low as 0.2 mg/kg.


Acute, subacute, and residual effects of polychlorinated biphenyl (pcb) in rats. II. Pathology and electron microscopy of liver and serum enzyme study.
This study was undertaken to determine the residual effects of a polychlorinated biphenyl (Aroclor 1254) fed to male rats at dietary concentrations of 0, 5, 50, and 500 ppm in diet. The animals were treated for 4 wk (acute and subacute phase), then observed for periods of up to 50 wk following termination of exposure (residual phase). The most significant histopathologic alteration was fatty degenerative change in the liver, which was most marked at 9 wk. Forty-six weeks postexposure, more than 50% of the rats fed 500 ppm still demonstrated fatty degenerative changes. On electron microscopic examination, marked increases in lipid vacuoles and smooth endoplasmic reticulum (SER) occurred with a concomitant decrease in rough endoplasmic reticulum (RER) profiles in the animals receiving 50-500ppm for 4 wk. Thirty-seven weeks following the exposure period, rats dosed with 50 ppm showed partial recovery toward control morphology (less lipid, less SER, more RER), while those receiving 500 ppm did not. Persistent morphologic alterations included an increase in SER and medium-density lipid material within cisternae of Ser, Golgi and Golgi-condensing vesicles, as well as a decrease in parallel arrays of RER. The persistence of ultrastructural alteration throughout the 46-wk residual phase emphasizes the long-lasting effects of 4-wk exposure to polychlorinated biphenyl.


Data mining and statistically guided clinical review of adverse event data in clinical trials.
Some approaches to the analysis of adverse event data arising from clinical trials are presented. These include (a) an inside-out data mining method where the adverse events are used as explanatory variables, classifying the treatment allocation, (b) a support method where we fit separate regression models to each adverse event with and without a treatment effect, and (c) a three-level hierarchical Bayesian mixture model for analysis of adverse event counts. The problem of understanding treatment-emergence of the adverse events is formulated as one of data mining rather than hypothesis testing. Our approaches provide an ordering of the adverse events by the strength of evidence of a treatment effect, rather than p values for prespecified hypotheses. The three methods produce intuitive graphical summaries showing the treatment effect on adverse event incidence. These graphs can be readily linked to relevant supportive information such as reports summarizing predicted risks for (demographic) subpopulations of interest and patient-level data such as laboratory information, concomitant medications, and medical history. This results in a statistically guided and thorough review of drug safety in the clinical trial.


DNA adsorption at functionalized Si/buffer interfaces studied by x-ray reflectivity.
The adsorption of DNA on chemically homogeneous, functionalized, oxide-free single-crystal silicon surfaces is studied by x-ray reflectivity. The adsorption of monodisperse, 294 base-pair double-stranded DNA on a positively charged surface is detected through the deformation of the molecular monolayer of aminated alkyl-chain molecules covalently bonded to the surface. The adsorption of single-stranded DNA does not lead to the same deformation. A detailed quantitative characterization of the density profiles yield surface densities of the covalently grafted, molecular monolayers that are in excellent agreement with infrared spectroscopic measurements. The additional mass density that is measured following the adsorption of DNA corresponds either to the partial embedding of a densely-packed adsorbed layer or to a deeper penetration into the soft surface layer at a lower surface density of the adsorbed double-stranded DNA molecules. The adsorption is found to be irreversible under high added salt concentrations, suggesting a partial dehydration of the double-stranded DNA.


Bacterial uropathogens in urinary tract infection and antibiotic susceptibility pattern in jimma university specialized hospital, southwest ethiopia.
Urinary tract infection (UTI) is one of the most common bacterial infections encountered by clinicians in developing countries. Area-specific monitoring studies aimed to gain knowledge about the type of pathogens responsible for urinary tract infections and their resistance patterns may help the clinician to choose the correct empirical treatment. Therefore, the aim of this study was to determine the type and antibiotic resistance pattern of the urinary pathogens isolated from patients attending Jimma University Specialized Hospital from April to June 2010. A hospital based cross sectional stud was conducted and urine samples were collected using the mid-stream "clean catch" method from 228 clinically-suspected cases of urinary tract infections and tested bacteriologically using standard procedures. Antimicrobial susceptibility test was performed for the isolated pathogens using Kirby-Bauer disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. Significant bacteria were detected from 9.2% of the total patients. The most common pathogens isolated were Escherichia coli (33.3%), Klebsiella pneumoniae (19%) and S. saprophyticus (14.3%). E. coli and Klebsiella pneumoniae showed the highest percentage of resistance to ampicillin and amoxacillin (100%) however, all isolates of E. coli and K. pneumoniae were susceptible to ciprofloxacin. S. saprophyticus and S. aureus were resistant to ampicillin (100%) and amoxicillin (66.7%). For all UTI isolates, least resistance was observed against drugs such as ceftriaxone, gentamycin and chloramphenicol. This study finding showed that E. coli isolates were the predominant pathogens and the presence of bacterial isolates with very high resistance to the commonly prescribed drugs that in turn leaves the clinicians with very few alternative options of drugs for the treatment of UTIs. As drug resistance among bacterial pathogens is an evolving process, routine surveillance and monitoring studies should be conducted to provide physicians knowledge on the updated and most effective empirical treatment of UTIs.


Russell W Brain and the aetiology of multiple sclerosis--a historical perspective.
The contribution of British neurologist Russell Walter Brain (1895-1966) to the field of neurology is difficult to overestimate and his seminal work continues to influence modern neurological education and practice. In a landmark review published in the Quarterly Review of Medicine in 1930, he gives a critically important account summarising ideas of the time thought to underlie the then called 'disseminated sclerosis', a disease he notes to be, 'after syphilis, the most frequent disease of the nervous system' in the UK. Across a century and a half, vast progress has been made in attempting to elucidate the as yet unknown cause of MS, which is unravelling to be multifactorial, highly complex and likely dependent on both genetic and environmental risk factors. Brain's observations highlight the changing epidemiology of MS over the last century which are likely to provide the platform in striving towards elucidating MS causation, notably a seemingly reduced latitudinal gradient of MS incidence, an increasing female-to-male sex ratio and an increasing disease rate in dark-skinned compared to light-skinned individuals. In this report we aim to evaluate the relevance today of what we believe to have been an important review demonstrating a perspective on MS far ahead that of its time, with a focus on Brain's ideas on the aetiology of MS; many of which have stood the test of time.


ICln, a novel integrin alphaIIbbeta3-associated protein, functionally regulates platelet activation.
A critical role for the conserved alpha-integrin cytoplasmic motif, KVGFFKR, is recognized in the regulation of activation of the platelet integrin alpha(IIb)beta(3). To understand the molecular mechanisms of this regulation, we sought to determine the nature of the protein interactions with this cytoplasmic motif. We used a tagged synthetic peptide, biotin-KVGFFKR, to probe a high density protein expression array (37,200 recombinant human proteins) for high affinity interactions. A number of potential integrin-binding proteins were identified. One such protein, a chloride channel regulatory protein, ICln, was characterized further because its affinity for the integrin peptide was highest as was its expression in platelets. We verified the presence of ICln in human platelets by PCR, Western blots, immunohistochemistry, and its co-association with alpha(IIb)beta(3) by surface plasmon resonance. The affinity of this interaction was 82.2 +/- 24.4 nm in a cell free assay. ICln co-immunoprecipitates with alpha(IIb)beta(3) in platelet lysates demonstrating that this interaction is physiologically relevant. Furthermore, immobilized KVGFFKR peptides, but not control KAAAAAR peptides, specifically extract ICln from platelet lysates. Acyclovir (100 microm to 5 mm), a pharmacological inhibitor of the ICln chloride channel, specifically inhibits integrin activation (PAC-1 expression) and platelet aggregation without affecting CD62 P expression confirming a specific role for ICln in integrin activation. In parallel, a cell-permeable peptide corresponding to the potential integrin-recognition domain on ICln (AKFEEE, 10-100 microm) also inhibits platelet function. Thus, we have identified, verified, and characterized a novel functional interaction between the platelet integrin and ICln, in the platelet membrane.


Application of alicyclic beta-amino acids in peptide chemistry.
The self-organizing beta-peptides have attracted considerable interest in the fields of foldamer chemistry and biochemistry. These compounds exhibit various stable secondary structure motifs that can be exploited to construct biologically active substances and nanostructured tertiary structures. The secondary structures can be controlled via the beta-amino acid sequence, and cyclic beta-amino acid residues play a crucial role in the design. The most important procedures for the preparation of cyclic beta-amino acid monomers and peptides are discussed in this tutorial review. Besides the secondary structure design principles, the methods of folded structure detection are surveyed.


Th17 cells undergo Fas-mediated activation-induced cell death independent of IFN-gamma.
IL-17-secreting CD4+ T cells (Th17 cells) play a critical role in immune responses to certain infections and in the development of many autoimmune disorders. The mechanisms controlling homeostasis in this cell population are largely unknown. In this study, we show that murine Th17 cells undergo rapid apoptosis in vitro upon restimulation through the TCR. This activation-induced cell death (AICD), a common mechanism for elimination of activated T cells, required the Fas and FasL interaction: Fas was stably expressed, while FasL was up-regulated upon TCR reactivation of Th17 cells; Ab ligation of Fas induced Th17 cell death; and AICD was completely absent in Th17 cells differentiated from gld/gld CD4+ T cells. Thus, the Fas/FasL pathway is essential in regulating the AICD of Th17 cells. Interestingly, IFN-gamma, a cytokine previously found to be important for the AICD of T cells, did not affect Th17 cell apoptosis. Furthermore, Th17 cells derived from mice deficient in IFN-gamma receptor 1 (IFN-gammaR1-/-) underwent AICD similar to wild-type cells. Thus, AICD of Th17 cells occurs via the Fas pathway, but is independent of IFN-gamma.


Comparison of pulsed-field ablation versus very high power short duration-ablation for pulmonary vein isolation.
The newly introduced nonthermal pulsed field ablation (PFA) is a promising technology to achieve fast pulmonary vein isolation (PVI) with high acute success rates and good safety features. However, previous studies have shown that very high power short duration ablation (VHPSD) is also highly effective and fast to achieve PVI with potentially less arrhythmia recurrence compared to conventional radiofrequency ablation. Data comparing PFA to VHPSD-PVI is lacking. This study compared procedural and outcome data for PFA-PVI to VHPSD-PVI in patients with paroxysmal or persistent atrial fibrillation (PAF/persAF). Consecutive patients undergoing de novo PVI (PFA or VHPSD) were included in this analysis. For PFA-PVI a pentaspline 20 electrode catheter was used. For VHPSD-PVI an enhanced irrigated catheter with a power setting of 70 W/7 s (70 W/5 s at posterior wall) was employed in conjunction with electro-anatomical mapping. All procedures were performed in deep analgo-sedation. A total of n = 114 patients (n = 57[50%] PFA, n = 17[30%] PAF; n = 40[70%] persAF) were included in this analysis. PVI was successful in all patients. The PFA group revealed a significantly shorter procedure duration (65 ± 17 min vs. 95 ± 23 min, p < 0.01) but longer fluoroscopy time (PFA 15 ± 5 min and VHPSD 12 ± 3 min; p < 0.001). At follow-up after median 125 days (interquartile range: 109-162) n = 46 PFA (80.7%) and n = 44 VHPSD pts (77.2%) were free from atrial arrhythmia after a single procedure (p = 0.819). Two tamponades occurred in the PFA while in VHPSD two pts suffered groin bleedings. One clinically nonsignificant PV stenosis occurred in the VHPSD group. Pulsed-field ablation and VHPSD-PVI seem to be highly effective and safe to achieve PVI in the setting of PAF and persAF with comparable arrhythmia-free survival. However, procedure duration for PFA PVI is significantly shorter and therefore may be of potential benefit. Compared to PFA VHPSD-PVI might ensure information on left atrial substrate allowing to target concomitant secondary tachycardias.


Bone Formation Following Sinus Augmentation with an Equine-Derived Bone Graft: A Retrospective Histologic and Histomorphometric Study with 36-Month Follow-up.
The aim of this study was to investigate bone formation over time following maxillary sinus augmentation with an enzyme-deantigenic, bone collagen-preserving equine bone graft by retrospective assessment of histomorphometric data. Records of patients with atrophic ridges who underwent maxillary sinus augmentation with the enzyme-deantigenic equine bone graft and two-step implant placement between 3 and 12 months after the sinus-augmentation surgery were assessed retrospectively. The histomorphometric data were clustered in three classes according to time of collection from the augmentation surgery and analyzed to assess newly formed bone deposition and residual biomaterial degradation rates. Data concerning the 36-month clinical follow-up were also assessed. Records of 77 patients and 115 biopsy specimens were retrieved, and histomorphometric data were clustered (3 to 5 months, n = 33; 6 to 8 months, n = 57; 9 to 12 months, n = 25). Mean minimum atrophic ridge thickness was 4.9 ± 0.5 mm (range, 4.0 to 7.1 mm). The amount of newly formed bone and residual biomaterial did not significantly differ among the three clusters. Qualitative analysis showed a denser trabecular structure in late (> 8 months) samples. At the 36-month clinical follow-up, no differences were found among the implant success rates in the three groups, according to the Albrektsson and Zarb criteria for success. The overall implant success rate was 98.3%. Based upon this retrospective human study of 77 patients with 4 to 7 mm of residual bone, when enzyme-deantigenic equine bone is used for sinus augmentation, new bone formation occurs at an early time (< 3 months) after the grafting, and implant placement can be safely carried out as soon as 3 to 5 months after the augmentation surgery.


Lexical Processing Strongly Affects Reading Times But Not Skipping During Natural Reading.
In a typical text, readers look much longer at some words than at others, even skipping many altogether. Historically, researchers explained this variation via low-level visual or oculomotor factors, but today it is primarily explained via factors determining a word's lexical processing ease, such as how well word identity can be predicted from context or discerned from parafoveal preview. While the existence of these effects is well established in controlled experiments, the relative importance of prediction, preview and low-level factors in natural reading remains unclear. Here, we address this question in three large naturalistic reading corpora (n = 104, 1.5 million words), using deep neural networks and Bayesian ideal observers to model linguistic prediction and parafoveal preview from moment to moment in natural reading. Strikingly, neither prediction nor preview was important for explaining word skipping-the vast majority of explained variation was explained by a simple oculomotor model, using just fixation position and word length. For reading times, by contrast, we found strong but independent contributions of prediction and preview, with effect sizes matching those from controlled experiments. Together, these results challenge dominant models of eye movements in reading, and instead support alternative models that describe skipping (but not reading times) as largely autonomous from word identification, and mostly determined by low-level oculomotor information.


[Effect of combined and local cytokine- and ozone therapy on the indices of lipid peroxidation, endogenous intoxication and ferroproteins in diffuse peritonitis].
The article analyzes the results of effect of combined and local cytokine- and ozone therapy on the indices of lipid peroxidation, endogenous intoxication and ferroproteins in 111 patients with diffuse peritonitis. It was shown, that combined sequential local and systemic cytokine and ozone therapy allows correcting the expression of endogenous intoxication and lipid peroxidation in diffuse peritonitis. This method suppresses an inflammation in the abdominal cavity. At the same time, it accelerates the elimination of intestine atony and thereby potentiates the possibilities of traditional methods of treatment.


Dendrimer-conjugated glutaminase inhibitor selectively targets microglial glutaminase in a mouse model of Rett syndrome.
Background: Elevated glutamate production and release from glial cells is a common feature of many CNS disorders. Inhibitors of glutaminase (GLS), the enzyme responsible for converting glutamine to glutamate have been developed to target glutamate overproduction. However, many GLS inhibitors have poor aqueous solubility, are unable to cross the blood brain barrier, or demonstrate significant toxicity when given systemically, precluding translation. Enhanced aqueous solubility and systemic therapy targeted to activated glia may address this challenge. Here we examine the impact of microglial-targeted GLS inhibition in a mouse model of Rett syndrome (RTT), a developmental disorder with no viable therapies, manifesting profound central nervous system effects, in which elevated glutamatergic tone, upregulation of microglial GLS, oxidative stress and neuroimmune dysregulation are key features. Methods: To enable this, we conjugated a potent glutaminase inhibitor, N-(5-{2-[2-(5-amino-[1,3,4]thiadiazol-2-yl)-ethylsulfanyl]-ethyl}-[1,3,4]thiadiazol-2-yl)-2-phenyl-acetamide (JHU29) to a generation 4 hydroxyl PAMAM dendrimer (D-JHU29). We then examined the effect of D-JHU29 in organotypic slice culture on glutamate release. We also examined GLS activity in microglial and non-microglial cells, and neurobehavioral phenotype after systemic administration of D-JHU29 in a mouse model of RTT. Results: We report successful conjugation of JHU29 to dendrimer resulting in enhanced water solubility compared to free JHU29. D-JHU29 reduced the excessive glutamate release observed in tissue culture slices in a clinically relevant Mecp2-knockout (KO) RTT mouse. Microglia isolated from Mecp2-KO mice demonstrated upregulation of GLS activity that normalized to wild-type levels following systemic treatment with D-JHU29. Neurobehavioral assessments in D-JHU29 treated Mecp2-KO mice revealed selective improvements in mobility. Conclusion: These findings demonstrate that glutaminase inhibitors conjugated to dendrimers are a viable mechanism to selectively inhibit microglial GLS to reduce glutamate production and improve mobility in a mouse model of RTT, with broader implications for selectively targeting this pathway in other neurodegenerative disorders.


Indole Alkaloid Derivative B, a Novel Bifunctional Agent That Mitigates 5-Fluorouracil-Induced Cardiotoxicity.
Clinically approved therapeutics that mitigate chemotherapy-induced cardiotoxicity, a serious adverse effect of chemotherapy, are lacking. The aim of this study was to determine the putative protective capacity of a novel indole alkaloid derivative B (IADB) against 5-fluorouracil (5-FU)-induced cardiotoxicity. To assess the free-radical scavenging activities of IADB, the acetylcholine-induced relaxation assay in rat thoracic aorta was used. Further, IADB was tested in normal and cancer cell lines with assays gauging autophagy induction. We further examined whether IADB could attenuate cardiotoxicity in 5-FU-treated male ICR mice. We found that IADB could serve as a novel bifunctional agent (displaying both antioxidant and autophagy-modulating activities). Further, we demonstrated that IADB induced production of cytosolic autophagy-associated structures in both cancer and normal cell lines. We observed that IADB cytotoxicity was much lower in normal versus cancer cell lines, suggesting an enhanced potency toward cancer cells. The cardiotoxicity induced by 5-FU was significantly relieved in animals pretreated with IADB. Taken together, IADB treatment, in combination with chemotherapy, may lead to reduced cardiotoxicity, as well as the reduction of anticancer drug dosages that may further improve chemotherapeutic efficacy with decreased off-target effects. Our data suggest that the use of IADB may be therapeutically beneficial in minimizing cardiotoxicity associated with high-dose chemotherapy. On the basis of the redox status difference between normal and tumor cells, IADB selectively induces autophagic cell death, mediated by reactive oxygen species overproduction, in cancer cells. This novel mechanism could reveal novel therapeutic targets in chemotherapy-induced cardiotoxicity.


Key items for reports of primary care research: an international Delphi study.
Reporting guidelines can improve dissemination and application of findings and help avoid research waste. Recent studies reveal opportunities to improve primary care (PC) reporting. Despite increasing numbers of guidelines, none exists for PC research. This study aims to prioritise candidate reporting items to inform a reporting guideline for PC research. Delphi study conducted by the Consensus Reporting Items for Studies in Primary Care (CRISP) Working Group. International online survey. Interdisciplinary PC researchers and research users. We drew potential reporting items from literature review and a series of international, interdisciplinary surveys. Using an anonymous, online survey, we asked participants to vote on and whether each candidate item should be included, required or recommended in a PC research reporting guideline. Items advanced to the next Delphi round if they received>50% votes to include. Analysis used descriptive statistics plus synthesis of free-text responses. 98/116 respondents completed round 1 (84% response rate) and 89/98 completed round 2 (91%). Respondents included a variety of healthcare professions, research roles, levels of experience and all five world regions. Round 1 presented 29 potential items, and 25 moved into round 2 after rewording and combining items and adding 2 new items. A majority of round 2 respondents voted to include 23 items (90%-100% for 11 items, 80%-89% for 3 items, 70%-79% for 3 items, 60%-69% for 3 items and 50%-59% for 3 items). Our Delphi study identified items to guide the reporting of PC research that has broad endorsement from the community of producers and users of PC research. We will now use these results to inform the final development of the CRISP guidance for reporting PC research.


Role of lymph flow in intestinal chylomicron transport.
In this study we investigated the influence of lymph flow on chylomicron transport. We examined the effects of varying the hydration of the interstitial matrix on chylomicron appearance time and on lymphatic lipid transport rate when a lipid test meal containing oleic acid and 1-monoolein was infused intraduodenally at a constant rate. The three groups of rats tested were control rats (normal interstitial hydration), rats receiving intravenous saline infusion (expanded interstitial matrix), and rats with an attenuated water absorption rate (dehydrated interstitial matrix). This study shows that lymph flow has a profound effect on intestinal chylomicron transport. As lymph flow increased, the chylomicron appearance time (time between the placement of radioactive fatty acid into the intestinal lumen to the appearance of radioactive lipid in the central lacteal) was reduced. When lymph flow exceeded 40 microliter/min, the chylomicron appearance time reached a minimum value of 13.6 min. This minimum chylomicron appearance time probably represents the time required for assembly of absorbed lipid, formation of chylomicrons, and their subsequent discharge into the lymphatics. The chylomicron appearance time lengthened as lymph flow fell. The results of this study underscore the necessity of using steady-state lymphatic lipid output data to assess factors affecting the cellular packaging and release of chylomicrons in the small intestine.


Managing advanced prostate cancer: the rapidly changing treatment landscape.
The FDA approval of 5 new therapeutic agents within a 4-year time frame, all of which potentially improve survival for patients with metastatic castration-resistant prostate cancer, is unprecedented in oncology. The diversity of the mechanisms of action of these agents is compelling, including the first and only therapeutic cancer vaccine, the first approved use of an alpha emitter, 2 novel next generation hormonal therapy agents, and a second-generation taxane. However, with this therapeutic bounty comes new challenges in the optimal use of these new and expensive agents in a disease well known for its heterogeneity.


[Comparative study of light and electron microscopic aspects of myocardial lesions induced in the rat by oils with several amounts of gamma-linolenic acid (author's transl)].
Four different oils : onager oil, mixture of onager oil (20 p. cent) and middle class triglyceride, sunflower oil and soybean oil were given to the rat during three different periods of time : 1) during 3 months after weaning ; 2) during 6 months after weaning ; 3) from the 9th to 15th months. Three types of myocardial lesions were found : interstitial inflammatory reaction, foci of cellular necrosis and fibrosis. The semi-quantitative study of these lesions did not find important differences from one oil to the other, as well as with the controls. However, subgroups onager are among the least altered, particularly as far as necrotic lesions are concerned. Morphometry of mitochondria, despite evident alterations under electron microscope, gave similar results in every groups.


Involvement of the transcription factor twist in phenotype alteration through epithelial-mesenchymal transition in lung cancer cells.
Epithelial-mesenchymal transition (EMT), which involves the persistent loss of epithelial markers and expression of mesenchymal markers, is assumed to have a critical role in not only tissue development during embryogenesis but also central mechanisms that enhance the invasive and metastatic ability of cancer cells. Twist has been identified to play an essential role in EMT-mediated tumor invasion and metastasis. Although recent studies suggest that twist expression levels in tissue specimens of lung cancer might be associated with prognosis, the expression of twist in lung cancer cells itself and its effect have not been fully evaluated. Here, we evaluated twist expression and its effect on phenotype alteration in lung cancer cell lines. Twist expression varied among human lung cancer cell lines. The lung cancer cell lines with high twist expression also tended to show a high vimentin/E-cadherin ratio, which was supported by a migration assay, in which high twist expression gave rise to high cell motility. Furthermore, in comparison to control cells, the lung cancer cells with ectopic expression of twist showed a significant phenotype alteration through EMT and an increasing ability to migrate in vitro, in part, due to a tenfold increase in matrix metalloproteinases activity and almost a 60% increase in modulation of focal adhesion kinase activity, although a contribution of microRNA appeared unlikely in our study. Our present analysis of twist expression in lung cancer provide clues to comprehensive understanding of the mechanisms, by which metastasis often develops in lung cancer.


[Simplification of methods for the study of beta hemolytic Streptococcus in faryngeal exudate].
A procedure of possible epidemiological application is described. The material is taken from pharynx in infants in an attempt to control the serious problem of rheumatic fever in our environment. The investigation is carried out in two steps: in the first, the samples are inoculated in a selective medium impregnated in filter paper strips, from where they are transferred to blood agar plates where 15 to 20 inoculations may be practiced; with this, unnecessary waste of individual plates is avoided, as done following conventional methods. The equipment may be easily adapted for direct takings.


Interleukin-4 (IL-4) pathway.
Interleukin-4 (IL-4) is a cytokine produced by T(H)2 type helper T cells and by mast cells, basophils, and eosinophils. This cytokine can elicit many responses, some of which are associated with allergy and asthma. Studies with long-term cell lines and primary cells have revealed differences in the signaling between these two experimental systems. Understanding these differences is important because therapeutic strategies targeting IL-4 and its signaling pathways are currently being tested to treat allergy and asthma.


Lethals in finite populations.
It has been assumed, based on theoretical studies, that lethals with the level of dominance estimated from experimental studies would have an allele frequency that is virtually independent of effective population size. However, here it is shown numerically that the expected frequency of lethals with low levels of dominance is also dependent on finite population size, although not as much as completely recessive lethals. This finding is significant in determining the standing level of inbreeding depression and the consequent potential for the evolution of self-fertilization. In addition, the architecture of genetic variation influencing inbreeding depression in populations with a history of small size may be of important consequence in endangered species. Finally, it is shown that the loss of lethal genetic variation often occurs much more quickly than the regeneration of lethal variation by mutation. This asymmetry may result in a lower standing genetic variation for inbreeding depression than expected from mutation rates and contemporary population size data.


Epithelial and mucosal preparations from canine colon: responses to substance P.
The hypothesis that substance P (SP) elicits both direct and indirect responses on the canine proximal colon was tested using two different in vitro preparations. The mucosa contained the muscularis mucosa and the attendant submucosal plexuses whereas the epithelium, being devoid of both, was functionally "nerve-free." Dose-dependent stimulation was noted on both preparations, increases in peak current (microamperes per squared centimeter) as well as charge transfers (millicoulombs per squared centimeter) were monitored. Tetrodotoxin significantly reduced mucosal responses whereas atropine and the H1 antagonists mepyramine and diphenhydramine had only marginal effects. None of these agents affected the responses of the epithelium to SP. Indomethacin significantly reduced responses in both preparations. Removal of Na+ or Cl- or the use of C- channel blockers (9-anthracene carboxylic acid and N-phenylanthranilic acid) produced a significant reduction of SP responses across the epithelium. Thus, SP has both direct and indirect affects on the colon; activation of the cyclooxygenase pathway could be significant.


Effect of a high copper intake and different levels of molybdenum on the health of sheep.
Molybdenum at levels of 2, 20, 40, and 60 mg/kg dry matter was added to a sheep ration containing 82 copper and 3, 8 g sulphur per kg dry matter. At the 2 higher levels of molybdenum, a substantial accumulation of copper and molybdenum was observed in the kidney cortices of the South African Mutton Merinos (reported elsewhere). The effect of these treatments on the health of the sheep was investigated. Observations on the 3 molybdenum supplemented treatments were: Serum enzyme levels remained normal throughout the trial. No evidence of kidney damage due to treatment could be obtained from kidney clearance studies. No pronounced liver lesions were evident from histophathological evaluations. A significant increase in Periodic acid Shiff positive stained granules was observed in the cortical tubules of the kidneys at increased molybdenum intakes. However, this was considered to be of minimal physiological significance. It was concluded that molybdenum prevented liver damage due to excess copper and that copper and molybdenum which did accumulate in the kidneys were probably harmless and present in an unavailable form to the body or caused minimal morphological changes. No haemolytic crisis occurred in the sheep receiving no added molybdenum. However, their concentration of serum enzymes Aspartate transaminase and Lactate dehydrogenase increased throughout the 193 day trial. It is suggested that elevated levels of these enzymes may be unreliable indicators of an approaching haemolytic crisis in the Merino.


Overview of exposure to and effects from radionuclides in terrestrial and marine environments.
The accident at the Fukushima Daiichi nuclear power plant, precipitated by the devastating earthquake and subsequent tsunami that struck the northeastern coast of Japan in March 2011, has raised concerns about potential impacts to terrestrial and marine environments from radionuclides released into the environment. A preliminary understanding of the potential ecological impacts from radionuclides can be ascertained from observations and data developed following previous environmental incidents elsewhere in the world. This article briefly summarizes how biota experience exposure to ionizing radiation, what effects may be produced, and how they may differ among taxa and habitats.


MicroRNA Transcriptome Profiling in Heart of Trypanosoma cruzi-Infected Mice: Parasitological and Cardiological Outcomes.
Chagas disease is caused by the parasite Trypanosoma cruzi, and it begins with a short acute phase characterized by high parasitemia followed by a life-long chronic phase with scarce parasitism. Cardiac involvement is the most prominent manifestation, as 30% of infected subjects will develop abnormal ventricular repolarization with myocarditis, fibrosis and cardiomyocyte hypertrophy by undefined mechanisms. Nevertheless, follow-up studies in chagasic patients, as well as studies with murine models, suggest that the intensity of clinical symptoms and pathophysiological events that occur during the acute phase of disease are associated with the severity of cardiac disease observed during the chronic phase. In the present study we investigated the role of microRNAs (miRNAs) in the disease progression in response to T. cruzi infection, as alterations in miRNA levels are known to be associated with many cardiovascular disorders. We screened 641 rodent miRNAs in heart samples of mice during an acute infection with the Colombiana T.cruzi strain and identified multiple miRNAs significantly altered upon infection. Seventeen miRNAs were found significantly deregulated in all three analyzed time points post infection. Among these, six miRNAs had their expression correlated with clinical parameters relevant to the disease, such as parasitemia and maximal heart rate-corrected QT (QTc) interval. Computational analyses identified that the gene targets for these six miRNAs were involved in networks and signaling pathways related to increased ventricular depolarization and repolarization times, important factors for QTc interval prolongation. The data presented here will guide further studies about the contribution of microRNAs to Chagas heart disease pathogenesis.


Natural Killer Cells and Dendritic Cells: Expanding Clinical Relevance in the Non-Small Cell Lung Cancer (NSCLC) Tumor Microenvironment.
Non-small cell lung cancer (NSCLC) is a major subtype of lung cancer that accounts for almost 85% of lung cancer cases worldwide. Although recent advances in chemotherapy, radiotherapy, and immunotherapy have helped in the clinical management of these patients, the survival rate in advanced stages remains dismal. Furthermore, there is a critical lack of accurate prognostic and stratification markers for emerging immunotherapies. To harness immune response modalities for therapeutic benefits, a detailed understanding of the immune cells in the complex tumor microenvironment (TME) is required. Among the diverse immune cells, natural killer (NK cells) and dendritic cells (DCs) have generated tremendous interest in the scientific community. NK cells play a critical role in tumor immunosurveillance by directly killing malignant cells. DCs link innate and adaptive immune systems by cross-presenting the antigens to T cells. The presence of an immunosuppressive milieu in tumors can lead to inactivation and poor functioning of NK cells and DCs, which results in an adverse outcome for many cancer patients, including those with NSCLC. Recently, clinical intervention using modified NK cells and DCs have shown encouraging response in advanced NSCLC patients. Herein, we will discuss prognostic and predictive aspects of NK cells and DC cells with an emphasis on NSCLC. Additionally, the discussion will extend to potential strategies that seek to enhance the anti-tumor functionality of NK cells and DCs.


An electrochlorination process integrating enhanced oxidation of phosphonate to orthophosphate and elimination: Verification of matrix chloridion-induced oxidation mechanism.
Phosphonate used as scale inhibitor is a non-negligible eutrophic contaminant in corresponding polluted waters. Besides, its conversion to orthophosphate (ortho-P) is a precondition for realizing bioavailable phosphorus recovery. Due to the feeble degradation efficiency with less than 30 % from classical Fenton commonly used in industrial wastewater treatment and itself vulnerable to strong inhibition interference of matrix chloride ions, we proposed an electrochemical approach to transform the native salt in the solution into oxidizing substances, sort of achieving beneficial utilization of matrix waste, and enhanced the ortho-P conversion rate of 1-Hydroxyethane-1,1-diphosphonic acid (HEDP) to 89.2 % (± 3.6 %). In electrochlorination system, it was found that HEDP rapidly complexed with Fe(II) and then coordinated in-situ Fe(III) to release free HEDP via intramolecular metal-ligand electron transfer reaction. The subsequent degradation mainly rooted in the oxidation of pivotal reactive species HClO, FeIVO2+ and 1O2, causing C-P and CC bonds to fracture in sequence. Eventually the organically bound phosphorus of HEDP was recovered as ortho-P. This study acquainted the audiences with the rare mechanism of chloridion-triggered HEDP degradation under electrochemical way, as well as offered a feasible technology for synchronous transformation of organically bound phosphorus to ortho-P and elimination from phosphonates.


Fitness facilitates sleep.
Eight army recruits were studied at the start, middle, and end of their initial 18-week training programme. At each point the subjects were studied for four consecutive nights in the sleep laboratory. Their sleep was characterized by the means of the recordings on the last two nights. Within 2 days of the sleep recordings (but never on the same day) each subject spent 2 non-consecutive days in the exercise laboratory. On the 1st day a maximum oxygen consumption (VO2 max) measurement was performed on a treadmill and on the 2nd day a 24-min progressive exercise bicycle ergometer test was carried out with simultaneous venous sampling (for lactic acid measurements) and oxygen consumption recordings from which the lactate turn point (LTP) was calculated. LTP was used as a measure of fitness. Approximately 1 week after the above measures lean muscle mass as calculated by total body potassium estimation was obtained for each subject. Slow wave sleep (SWS) as a percentage of total sleep time increased significantly between the start and the measurements at 9 and 18 weeks, being 21.9%, 29.9%, and 28.5% respectively. Anaerobic threshold increased significantly (P less than 0.05) over the first 9 weeks and continued to increase to the end of the training period (P less than 0.001) using VO2 when lactate level was 2 mmol/l as a percentage of VO2 max. With increase in fitness, sleep onset latency and wake time during sleep decreased and sleep efficiency improved. The results suggest that as fitness increases sleep quality improves.


DOC2B acts as a calcium switch and enhances vesicle fusion.
Calcium-dependent exocytosis is regulated by a vast number of proteins. DOC2B is a synaptic protein that translocates to the plasma membrane (PM) after small elevations in intracellular calcium concentration. The aim of this study was to investigate the role of DOC2B in calcium-triggered exocytosis. Using biochemical and biophysical measurements, we demonstrate that the C2A domain of DOC2B interacts directly with the PM in a calcium-dependent manner. Using a combination of electrophysiological, morphological, and total internal reflection fluorescent measurements, we found that DOC2B acts as a priming factor and increases the number of fusion-competent vesicles. Comparing secretion during repeated stimulation between wild-type DOC2B and a mutated DOC2B that is constantly at the PM showed that DOC2B enhances catecholamine secretion also during repeated stimulation and that DOC2B has to translocate to the PM to exert its facilitating effect, suggesting that its activity is dependent on calcium. The hypothesis that DOC2B exerts its effect at the PM was supported by the finding that DOC2B affects the fusion kinetics of single vesicles and interacts with the PM SNAREs (soluble NSF attachment receptors). We conclude that DOC2B is a calcium-dependent priming factor and its activity at the PM enables efficient expansion of the fusion pore, leading to increased catecholamine release.


Positionally isomeric organic gelators: structure-gelation study, racemic versus enantiomeric gelators, and solvation effects.
Low molecular weight gelator molecules consisting of aliphatic acid, amino acid (phenylglycine), and omega-aminoaliphatic acid units have been designed. By varying the number of methylene units in the aliphatic and omega-aminoaliphatic acid chains, as defined by descriptors m and n, respectively, a series of positionally isomeric gelators having different positions of the peptidic hydrogen-bonding unit within the gelator molecule has been obtained. The gelation properties of the positional isomers have been determined in relation to a defined set of twenty solvents of different structure and polarity and analyzed in terms of gelator versatility (G(ver)) and effectiveness (G(eff)). The results of gelation tests have shown that simple synthetic optimizations of a "lead gelator molecule" by variation of m and n, end-group polarity (carboxylic acid versus sodium carboxylate), and stereochemistry (racemate versus optically pure form) allowed the identification of gelators with tremendously improved versatility (G(ver)) and effectiveness (G(eff)). Dramatic differences in G(eff) values of up to 70 times could be observed between pure racemate/enantiomer pairs of some gelators, which were manifested even in the gelation of very similar solvents such as isomeric xylenes. The combined results of spectroscopic ((1)H NMR, FTIR), electron microscopy (TEM), and X-ray diffraction studies suggest similar organization of the positionally isomeric gelators at the molecular level, comprising parallel beta-sheet hydrogen-bonded primary assemblies that form inversed bilayers at a higher organizational level. Differential scanning calorimetry (DSC) studies of selected enantiomer/racemate gelator pairs and their o- and p-xylene gels revealed the simultaneous presence of different polymorphs in the racemate gels. The increased gelation effectiveness of the racemate compared to that of the single enantiomer is most likely a consequence of its spontaneous resolution into enantiomeric bilayers and their subsequent organization into polymorphic aggregates of different energy. The latter determine the gel fiber thickness and solvent immobilization capacity of the formed gel network.


Chronic myelogenous leukemia accompanied by megaloblastic anemia showing atypical clinical features.
Leukocytosis, splenomegaly, and an increased vitamin B(12) level are characteristic findings of chronic myelogenous leukemia in the chronic phase (CML-CP). Here, we report a patient with CML-CP accompanied by megaloblastic anemia. A 61-year-old man consulted our hospital because of anemia and thrombocytopenia. On physical examination, there were no remarkable findings; there was no hepatosplenomegaly. Laboratory findings were: hemoglobin 6.0 g/dl; MCV 113.6 fl; platelet count 100×10(9)/l; white cell count 8.66×10(9)/l; and LDH 1,236 IU/l. Peripheral blood smear demonstrated hypersegmented neutrophils and megalocytes with emergence of myeloblasts, giant metamyelocytes, and nucleated red cells. Vitamin B(12) and folic acid levels were low. Bone marrow examination showed megaloblastic change in the erythroblasts and myeloid hyperplasia. Following vitamin B(12) and folic acid administration, anemia and thrombocytopenia rapidly improved; thereafter, marked leukocytosis became evident. Based on the presence of t(9;22)(q34;q11) on cytogenetic study and a positive result for Major bcr/abl fusion gene, a diagnosis of CML-CP was established. This case illustrates that ineffective erythropoiesis results in anemia and thrombocytopenia in CML with vitamin B12 and/or folic acid deficiency.