- Single molecule TPM analysis of the catalytic pentad mutants of Cre and Flp site-specific recombinases: contributions of the pentad residues to the pre-chemical steps of recombination
- Fan, Hsiu-Fang, Cheng, Yong-Song, Ma, Chien-Hui, Jayaram, Makkuni | Nucleic acids research (v.43 / no.6 / pp.3237-3255 / 20150305-1048)
- Cre and Flp site-specific recombinase variants harboring point mutations at their conserved catalytic pentad positions were characterized using single molecule tethered p
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Cre and Flp site-specific recombinase variants harboring point mutations at their conserved catalytic pentad positions were characterized using single molecule tethered particle motion (TPM) analysis. The findings reveal contributions of these amino acids to the pre-chemical steps of recombination. They suggest functional differences between positionally conserved residues in how they influence recombinase-target site association and formation of #x2018;non-productive #x2019;, #x2018;pre-synaptic #x2019; and #x2018;synaptic #x2019; complexes. The most striking difference between the two systems is noted for the single conserved lysine. The pentad residues in Cre enhance commitment to recombination by kinetically favoring the formation of pre-synaptic complexes. These residues in Flp serve a similar function by promoting Flp binding to target sites, reducing non-productive binding and/or enhancing the rate of assembly of synaptic complexes. Kinetic comparisons between Cre and Flp, and between their derivatives lacking the tyrosine nucleophile, are consistent with a stronger commitment to recombination in the Flp system. The effect of target site orientation (head-to-head or head-to-tail) on the TPM behavior of synapsed DNA molecules supports the selection of anti-parallel target site alignment prior to the chemical steps. The integrity of the synapse, whose establishment/stability is fostered by strand cleavage in the case of Flp but not Cre, appears to be compromised by the pentad mutations.
- Sudden death associated with QT interval prolongation and KCNQ1 gene mutation in a family of English Springer Spaniels.
- Ware, W A, Reina-Doreste, Y, Stern, J A, Meurs, K M | Journal of veterinary internal medicine (v.29 / no.2 / pp.561-568 / 20150891-6640)
- A 5-year-old, healthy English Springer Spaniel died suddenly 4?months after delivering a litter of 7 puppies. Within 4?months of the dam #039;s death, 3 offspring also di
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A 5-year-old, healthy English Springer Spaniel died suddenly 4?months after delivering a litter of 7 puppies. Within 4?months of the dam #039;s death, 3 offspring also died suddenly.
- Survival or growth of inoculated Escherichia coli O157:H7 and Salmonella on yellow onions (Allium cepa) under conditions simulating food service and consumer handling and storage.
- Lieberman, Vanessa M, Zhao, Irene Y, Schaffner, Donald W, Danyluk, Michelle D, Harris, Linda J | Journal of food protection (v.78 / no.1 / pp.42-50 / 20150362-028x)
- Whole and diced yellow onions (Allium cepa) were inoculated with five-strain cocktails of rifampin-resistant Escherichia coli O157:H7 or Salmonella and stored under condi
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Whole and diced yellow onions (Allium cepa) were inoculated with five-strain cocktails of rifampin-resistant Escherichia coli O157:H7 or Salmonella and stored under conditions to simulate food service or consumer handling. The inoculum was grown in broth (for both whole and diced onion experiments) or on agar plates (for whole onion experiments). Marked circles (3.3 cm in diameter) on the outer papery skin of whole onions were spot inoculated (10 °ül in 10 drops) at 7 log CFU per circle, and onions were stored at 4 #176;C, 30 to 50 % relative humidity, or at ambient conditions (23 #176;C, 30 to 50 % relative humidity). Diced onions were inoculated at 3 log CFU/g and then stored in open or closed containers at 4 #176;C or ambient conditions. Previously inoculated and ambient-stored diced onions were also mixed 1:9 (wt/wt) with refrigerated uninoculated freshly diced onions and stored in closed containers at ambient conditions. Inoculated pathogens were recovered in 0.1 % peptone and plated onto selective and nonselective media supplemented with 50 °üg/ml rifampin. Both E. coli O157:H7 and Salmonella populations declined more rapidly on onion skins when the inoculum was prepared in broth rather than on agar. Agar-prepared E. coli O157:H7 and Salmonella declined by 0.4 and 0.3 log CFU per sample per day, respectively, at ambient conditions; at 4 #176;C the rates of reduction were 0.08 and 0.06 log CFU per sample per day for E. coli O157:H7 and Salmonella, respectively. Populations of E. coli O157:H7 and Salmonella did not change over 6 days of storage at 4 #176;C in diced onions. Lag times of 6 to 9 h were observed with freshly inoculated onion at ambient conditions; no lag was observed when previously inoculated and uninoculated onions were mixed. Growth rates at ambient conditions were 0.2 to 0.3 log CFU/g/h for E. coli O157:H7 and Salmonella in freshly inoculated onion and 0.2 log CFU/g/h in mixed product. Diced onions support pathogen growth and should be kept refrigerated.
- Current natural products with antihypertensive activity
- BAI, R.R., WU, X.M., XU, J.Y. | Chinese journal of natural medicines = ñéÏÐô¸æÔå·Úª (v.13 / no.10 / pp.721-729 / 20152095-6975)
- Natural products have been an important source of new drugs, which also played a dominant role in the discovery and research of new drugs for the treatment of hypertensio
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Natural products have been an important source of new drugs, which also played a dominant role in the discovery and research of new drugs for the treatment of hypertension. This review article reviews the recent progress in the research and development of natural lead compounds with antihypertensive activity, including alkaloids, diterpenes, coumarins, flavonoids, and peptides. We summarized their structures, sources, as well as the antihypertensive mechanisms. These information provides instructive reference for the following structural modifications and optimization.
- Steroid biosynthesis in adipose tissue
- Li, J., Papadopoulos, V., Vihma, V. | Steroids (v.103 / pp.89-104 / 20150039-128x)
- Tissue-specific expression of steroidogenic enzymes allows the modulation of active steroid levels in a local manner. Thus, the measurement of local steroid concentration
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Tissue-specific expression of steroidogenic enzymes allows the modulation of active steroid levels in a local manner. Thus, the measurement of local steroid concentrations, rather than the circulating levels, has been recognized as a more accurate indicator of the steroid action within a specific tissue. Adipose tissue, one of the largest endocrine tissues in the human body, has been established as an important site for steroid storage and metabolism. Locally produced steroids, through the enzymatic conversion from steroid precursors delivered to adipose tissue, have been proven to either functionally regulate adipose tissue metabolism, or quantitatively contribute to the whole body's steroid levels. Most recently, it has been suggested that adipose tissue may contain the steroidogenic machinery necessary for the initiation of steroid biosynthesis de novo from cholesterol. This review summarizes the evidence indicating the presence of the entire steroidogenic apparatus in adipose tissue and discusses the potential roles of local steroid products in modulating adipose tissue activity and other metabolic parameters.
- Natural products as inhibitors of recombinant cathepsin L of Leishmania mexicana
- de Sousa, L.R.F., Wu, H., Nebo, L., Fernandes, J.B., da Silva, M.F.d.G.F., Kiefer, W., Schirmeister, T., Vieira, P.C. | Experimental parasitology (v.156 / pp.42-48 / 20150014-4894)
- Cysteine proteinases (cathepsins) from Leishmania spp. are promising molecular targets against leishmaniasis. Leishmania mexicana cathepsin L is essential in the parasite
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Cysteine proteinases (cathepsins) from Leishmania spp. are promising molecular targets against leishmaniasis. Leishmania mexicana cathepsin L is essential in the parasite life cycle and a pivotal in virulence factor in mammals. Natural products that have been shown to display antileishmanial activity were screened as part of our ongoing efforts to design inhibitors against the L.@?mexicana cathepsin L-like rCPB2.8. Among them, agathisflavone (1), tetrahydrorobustaflavone (2), 3-oxo-urs-12-en-28-oic acid (3), and quercetin (4) showed significant inhibitory activity on rCPB2.8 with IC 50 values ranging from 0.43 to 18.03@?@?M. The mechanisms of inhibition for compounds 1-3, which showed K i values in the low micromolar range (K i @?=@?0.14-1.26@?@?M), were determined. The biflavone 1 and the triterpene 3 are partially noncompetitive inhibitors, whereas biflavanone 2 is an uncompetitive inhibitor. The mechanism of action established for these leishmanicidal natural products provides a new outlook in the search for drugs against Leishmania.
- SOLIDARITY in the Moral Imagination of Bioethics.
- Jennings, Bruce, Dawson, Angus | The Hastings Center report (v.45 / no.5 / pp.31-38 / 20150093-0334)
- How important is the concept of solidarity in our society #039;s calculus of consent as regards the legitimacy and ethical and political support for public health, health
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How important is the concept of solidarity in our society #039;s calculus of consent as regards the legitimacy and ethical and political support for public health, health policy, and health services? By the term 'calculus of consent,' we refer to the answer that people give to rationalize and justify their obedience to laws, rules, and policies that benefit others. The calculus of consent answers questions such as, Why should I care? Why should I help? Why should I contribute to the public provision of others? Consent here does not have to be a deliberate, explicit act of informed agreement. And a calculus does not have to be a quantifiable, quasimathematical operation; more often, such a calculus takes narrative form in stories that a society tells about itself and that individuals tell about their place in it. One vital function of bioethics is to inform and shape those stories. Bioethics has the potential to offer society a keener insight into and perception of what is ethically at stake in controversies concerning health, science, and society. This insight is what we shall refer to as a 'moral imagination,' by which we do not mean make-believe or fantasy but, rather, the capacity to take a critical distance from the given, to think reality otherwise. The moral imagination enables one to see connections between factors at work in history, in large social and communal structures, and in the shape of one #039;s own life, thoughts, and feelings. Here we are especially concerned with the contribution that the concept of solidarity can make to the moral imagination of bioethics. We contend that solidarity must become more widely active and explicit in bioethics analysis and argumentation as it endeavors to shape reasons for obeying norms and rules of common benefit in an open, diverse society.
- Relocation of CLIC1 Promotes Tumor Cell Invasion and Colonization of Fibrin
- Gurski, Lisa A., Knowles, Lynn M., Basse, Per H., Maranchie, Jodi K., Watkins, Simon C., Pilch, Jan | Molecular cancer research (v.13 / no.2 / pp.273-280 / 20151541-7786)
- Chloride intracellular channel 1 (CLIC1) has been shown to be upregulated in various malignancies but its exact function remains unclear. Here, it is revealed that CLIC1
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Chloride intracellular channel 1 (CLIC1) has been shown to be upregulated in various malignancies but its exact function remains unclear. Here, it is revealed that CLIC1 is critical for the stability of invadopodia in endothelial and tumor cells embedded in a 3-dimensional (3D) matrix of fibrin. Invadopodia stability was associated with the capacity of CLIC1 to induce stress fiber and fibronectin matrix formation following its #x03B2;3 integrin (ITGB3)-mediated recruitment into invadopodia. This pathway, in turn, was relevant for fibrin colonization as well as slug (SNAI2) expression and correlated with a significant role of CLIC1 in metastasis in vivo . Mechanistically, a reduction of myosin light chain kinase (MYLK) in CLIC1-depleted as well as #x03B2;3 integrin-depleted cells suggests an important role of CLIC1 for integrin-mediated actomyosin dynamics in cells embedded in fibrin. Overall, these results indicate that CLIC1 is an important contributor to tumor invasion, metastasis, and angiogenesis. Implications: This study uncovers an important new function of CLIC1 in the regulation of cell #x2013;extracellular matrix interactions and ability of tumor cells to metastasize to distant organs. Mol Cancer Res; 13(2); 273 #x2013;80. #x00A9;2014 AACR .
- Local allergic rhinitis in elderly patients
- Bozek, A., Ignasiak, B., Kasperska-Zajac, A., Scierski, W., Grzanka, A., Jarzab, J. | Annals of allergy, asthma, immunology (v.114 / no.3 / pp.199-202 / 20151081-1206)
- Background: Local allergic rhinitis (LAR) is characterized by the production of specific IgE in the nasal mucosa and a positive response to a nasal provocation test in th
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Background: Local allergic rhinitis (LAR) is characterized by the production of specific IgE in the nasal mucosa and a positive response to a nasal provocation test in the absence of atopy by conventional measurements. There is no information about LAR in elderly people, just as there is little information about allergic rhinitis (AR) in this age group. Objective: To investigate the prevalence of LAR, AR, and non-AR in elderly patients with rhinitis. Methods: In 219 patients with a mean (SD) age of 65.81 (5.88), skin prick tests, serum total specific IgE, and nasal provocation tests against common aeroallergens were performed. In addition, nasal specific IgE was measured in the nasal lavage at baseline and after provocation. For monitoring nasal symptoms, a visual analog scale was used. Results: Of the 219 patients, 46 (21.0%) had LAR, 88 (40.2%) had AR, and 85 (38.8%) were diagnosed as having non-AR. Dermatophagoides pteronyssinus was the main sensitizing aeroallergen in patients with LAR (29 patients [63.0%]) and with AR (48 patients [56.4%]). No significant differences were found between the visual analog scale score and the type of AR (local or nonlocal) and the types of allergens. Clinical responses during the nasal provocation test were associated with significant increases in nasal IgE (Spearman correlation test, R = 0.89, P
- Measuring the economic costs of discrimination experienced by people with mental health problems: development of the Costs of Discrimination Assessment (CODA).
- Wright, Steve, Henderson, Claire, Thornicroft, Graham, Sharac, Jessica, McCrone, Paul | Social psychiatry and psychiatric epidemiology (v.50 / no.5 / pp.787-795 / 20150933-7954)
- Stigma and discrimination are faced by many with mental health problems and this may affect the uptake of services and engagement in leisure and recreational activities.
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Stigma and discrimination are faced by many with mental health problems and this may affect the uptake of services and engagement in leisure and recreational activities. The aims of this study were to develop a schedule to measure the impact of stigma and discrimination on service use, employment and leisure activities and to estimate the value of such reductions.
- Parvovirus B19 myocarditis in children: an observational study
- Vigneswaran, Trisha V, Brown, Julianne R, Breuer, Judith, Burch, Michael | Archives of disease in childhood (v.101 / no.2 / pp.177-180 / 20160003-9888)
- Background The advent of PCR testing for the presence of viral genomes has led to the identification of parvovirus B19 (PVB19) as a causative agent of myocarditis. Meth
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Background The advent of PCR testing for the presence of viral genomes has led to the identification of parvovirus B19 (PVB19) as a causative agent of myocarditis. Methods The clinical presentation, course and outcome of children with PVB19 myocarditis was ascertained through a retrospective review. The PVB19 viral genome was detected by PCR from whole blood or endomyocardial biopsy specimens in patients presenting with new onset heart failure. Results Seventeen patients presented at a median age of 1.3 years (range: 0.4 #x2013;15.4 #x2005;years) in cardiac failure with a mean fractional shortening of 15 #x00B1;3%. Eleven patients required mechanical ventilation and intravenous inotropes and seven required extra-corporeal mechanical oxygenation. Four of the five deaths occurred in patients who had a short prodromal illness of less than 48 #x2005;hours. All patients with ST segment elevation died (n=4). All non-fulminant cases survived. Event-free survival occurred in 11/17 (65%) patients. Five (29%) patients died and one patient underwent heart transplantation. Complete recovery of cardiac function occurred within a median of 12 months (range: 1 #x2013;48) in five patients. There was incomplete recovery in five patients and one patient had persistent dilated cardiomyopathy. Conclusions PVB19 can cause a devastating myocarditis in children. Children with fulminant myocarditis, ST segment changes or a short prodrome have the worst outcome. Transplantation may be considered, but is rarely required in the acute period if mechanical circulatory support is utilised. If the initial presentation is survived, recovery of the myocardium can occur even in those who had fulminant myocarditis.
- Effects of oxygen impurities and nitrogen vacancies on the surface properties of the Ta3N5 photocatalyst: a DFT study
- Wang, J., Ma, A., Li, Z., Jiang, J., Feng, J., Zou, Z. | Physical chemistry chemical physics : PCCP (v.17 / no.35 / pp.23265-23272 / 20151463-9076)
- The effect of oxygen in Be- and Si-doped GaInP grown by all-solid-source molecular beam epitaxy is reported. The oxygen incorporation in Si-doped material is reduced rela
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- Clic1 plays a role in mouse hepatocarcinoma via modulating Annexin A7 and Gelsolin in vitro and in vivo
- Zhang, J., Li, M., Song, M., Chen, W., Mao, J., Song, L., Wei, Y., Huang, Y., Tang, J. | Biomedicine pharmacotherapy = Biom eacute;decine pharmacoth eacute;rapie (v.69 / pp.416-419 / 20150753-3322)
- Clic1 is a member of the family of chloride intracellular ion channels. Previous studies suggest that Clic1 is involved in migration and invasion of the lymphatic metasta
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Clic1 is a member of the family of chloride intracellular ion channels. Previous studies suggest that Clic1 is involved in migration and invasion of the lymphatic metastasis in hepatocarcinoma, however, the mechanism is not fully understood. In the present study, we observed Clic1 is abundant in cytoplasm, higher expression in Hca-F cell than Hca-P cell, and we showed that downregulation of Clic1 by RNA interference was able to markedly enhance the expression of tumor metastasis genes Annexin A7 and Gelsolin in vitro, and downregulation of Annexin A7 and Gelsolin also enhanced the expression of Clic1 in vitro and in vivo. Our results provide novel insight that Clic1 have a role in migration and invasion in hepatocarcinoma maybe via modulating the expression of Annexin A7 and Gelsolin, and provide novel insight into the mechanisms of Clic1 for hepatocarcinoma treatment.
- Phenotype-Guided Natural Products Discovery Using Cytological Profiling
- Ochoa, Jessica L., Bray, Walter M., Lokey, R. Scott, Linington, Roger G. | Journal of natural products (v.78 / no.9 / pp.2242-2248 / 20150163-3864)
- Phenotype-guided natural products discovery is emerging as a useful new discovery tool that addresses challenges in early, unbiased natural product biological annotation.
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Phenotype-guided natural products discovery is emerging as a useful new discovery tool that addresses challenges in early, unbiased natural product biological annotation. These high-content approaches yield screening results that report directly on the impact of test compounds on cellular processes in target organisms and can be used to predict the modes of action of bioactive constituents from primary screening data. In this study we explored the use of our recently implemented cytological profiling platform for the isolation of compounds with a specific, predefined mode of action, namely, induction of mitotic arrest. Screening of a microbially derived extract library revealed six extracts whose cytological profiles clustered closely with those of known antimitotic agents from the pure compound training set. Subsequent examination of one of these extracts revealed the presence of two separate bioactive constituents, each of which possessed a unique cytological profile. The first, diketopiperazine XR334 ( 3 ), recapitulated the observed antimitotic phenotype of the original extract, demonstrating that cytological profiling can be used for the targeted isolation of compounds with specific modes of action. The second, nocapyrone L ( 6 ), possessed a cytological profile that clustered with known calcium channel modulators, in line with previous published activities for this compound class, indicating that cytological profiling is a flexible and powerful platform for the de novo characterization of compound modes of action. Graphic Abstract ACS Electronic Supporting Info
- Combined EoS and elastic constraints models to predict thermodynamic properties for systems involving semi-crystalline polyolefins
- Bashir, M.A., Ali, M.A.h., Kanellopoulos, V., Seppala, J. | Fluid phase equilibria (v.388 / pp.107-117 / 20150378-3812)
- In the present study, an equation of state model (i.e., Sanchez-Lacombe equation of state-SL EoS) and an Activity coefficient model (i.e., UNIFAC-FV) are combined with el
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In the present study, an equation of state model (i.e., Sanchez-Lacombe equation of state-SL EoS) and an Activity coefficient model (i.e., UNIFAC-FV) are combined with elastic constraints models to assess the effect of polymer crystallinity on solubility of penetrants (e.g., alpha;-olefins, diluents, light gases, etc.) in polyolefins (i.e., binary and ternary mixtures). It is shown that the predictive capabilities of the employed solubility models can be significantly improved when combined with corresponding models accounting for the elastic constraints effects on polymer chains (i.e., Michaels-Hausslein, Banaszak modification of Michaels-Hausslein). More specifically, it is illustrated that for the SL EoS, the fraction of elastically effective chains (f) is the only adjustable parameter which has to be considered to accurately predict the solubility of penetrants in semi-crystalline polyolefins at conditions of industrial importance. In addition, such a combined model results in reduction of the adjustable parameters (i.e., binary interaction parameters, k ij ), particularly in multi-component mixtures. It is also depicted that the parameter f is a function of polymer crystallinity and temperature in a way that f increases with increasing crystallinity and decreasing temperature. Moreover, it is demonstrated that the incorporation of elastic constraints models in UNIFAC-FV improves its predictive capabilities by about 25%. Finally, a comprehensive sensitivity analysis is carried out to investigate the effect of parameter f on the predictive capabilities of the SL-EoS combined with elastic constraints models.
- Probing the chain segment mobility at the interface of semi-crystalline polylactide/clay nanocomposites
- Saiter, A., Delpouve, N., Dargent, E., Oberhauser, W., Conzatti, L., Cicogna, F., Passaglia, E. | European polymer journal (v.78 / pp.274-289 / 20160014-3057)
- The concept of Cooperative Rearranging Region (CRR), an efficient probe of the interaction level in nanocomposites, has been used in Poly(lactic acid) (PLA)-based composi
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The concept of Cooperative Rearranging Region (CRR), an efficient probe of the interaction level in nanocomposites, has been used in Poly(lactic acid) (PLA)-based composites with phyllosilicates prepared in the melt with organomontmorillonite (O-MMT) and using poly(butylene adipate-co-terephthalate) (PBAT) as coupling agent. The samples were crystallized from various thermal treatments in order to obtain a wide range of morphologies and microstructures and accurately characterized by XRD, TEM, standard DSC and MT-DSC with the aim to highlight the effect of lamellae dispersion and distribution at nanoscale onto the thermal features of resulting nanocomposites. The presence of different interaction levels at the interface PLA/O-MMT, even tuned by the presence of PBAT, affects both the crystalline phase structure (by differently promoting crystallization of alpha; and alpha;' forms) and the distribution between the amorphous fractions (rigid and mobile). The variations of molecular dynamics are classified in two categories depending on the composite microstructure. Only in amorphous materials the cooperativity is driven by the filler/matrix interactions. In semi-crystalline materials, the morphological features linked to the presence of O-MMT and PBAT are overwhelmed by the confinement of the amorphous phase. Comparison between the two crystallization modes evidences a stronger change in the glass transition dynamics for systems exhibiting preponderant nucleation and high percentage of rigid amorphous fraction.