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TitleDate Created
An iterative O-methyltransferase catalyzes 1,11-dimethylation of Aspergillus fumigatus fumaric acid amides.Thursday, July 21, 2016
Kalb D, Heinekamp T, Schieferdecker S, Nett M, Brakhage A, Hoffmeister D,
Chembiochem : a European journal of chemical biology. 21-Jul-2016
S-adenosyl-L-methionine (SAM) dependent methyltransfer is a common biosynthetic strategy to modify natural products. The previously uncharacterized Aspergillus fumigatus methyltransferase FtpM was investigated, which is encoded next to the bimodular fumaric acid amide synthetase FtpA. Structure elucidation of two new A. fumigatus natural products, the 1,11-dimethyl esters of fumaryl-L-tyrosine and fumaryl-L-phenylalanine, together with ftpM gene disruption suggested that FtpM catalyzes iterative methylation. Final evidence that a single enzyme repeatedly acts on fumaric acid amides came from an in vitro biochemical investigation of recombinantly produced FtpM. Size exclusion chromatography indicated that this methyltransferase is active as a dimer. As ftpA and ftpM homologues are found clustered in other fungi as well, we expect our work helps identify and annotate natural product biosynthesis genes in various species.
Advances in understanding the mechanisms of erythropoiesis in homeostasis and disease.Thursday, July 21, 2016
Liang R, Ghaffari S,
British journal of haematology. 21-Jul-2016
Anaemia or decreased blood haemoglobin is the most common blood disorder often characterized by reduced red blood cell (RBC) numbers. RBCs are produced from differentiation and commitment of haematopoietic stem cells to the erythroid lineage by a process called erythropoiesis. Coordination of erythropoietin receptor signalling with several erythroid transcription factors including GATA1 is essential for this process. A number of additional players that are critical for RBC production have been identified in recent years. Major technological advances, such as the development of RNA interference, genetically modified animals, including zebrafish, and imaging flow cytometry have led to these discoveries; the emergence of -omics approaches in combination with the optimization of ex vivo erythroid cultures have also produced a more comprehensive understanding of erythropoiesis. Here we summarize studies describing novel regulators of erythropoiesis that modulate erythroid cell production in the context of human erythroid disorders involving hypoxia, iron regulation, immune-related molecules, and the transcription factor FOXO3.
Multi-constituent identification in Australian cane toad skin extracts using high-performance liquid chromatography high-resolution tandem mass spectrometry.Thursday, July 21, 2016
Zulfiker AH, Sohrabi M, Qi J, Matthews B, Wei MQ, Grice ID,
Journal of pharmaceutical and biomedical analysis. 1-Jul-2016
Toad skins and venom glandular secretions have been widely used for centuries in traditional Chinese and Japanese medicine for the treatment of various ailments such as cancer, sores, toothache, local inflammation and pain. The active chemical constituents from traditional oriental medicines have demonstrated potential in the development of effective therapeutic pharmaceuticals. Our primary focus in this research was to identify and characterise 'active' compounds or groups of compounds for their potential as neuropsychiatric disorder therapeutics. For this aim, we utilised a variety of solvents, i.e., the aqueous, 60% ethanol (aqueous) and acetic acid (aq) (at two different pHs) for extractions of Australian cane toad skins to identify chemical constituents. The identification of compounds was carried out using HPLC-ESI-Q-TOF-MS/MS based on the accurate mass measurement for molecular ions and MS/MS analysis, whereby accurate mass pseudo-molecular ions and characteristic fragment ions were compared to published reference data, including mass bank and NIST. As a result, we have to date identified 42 major constituents including alkaloids, amino acids, bufadienolides, fatty acids, nucleobases, nucleosides and vitamins mostly from the aqueous and 60% ethanol extracts. Of the 42 constituents identified, 29 were found in the aqueous extract, 35 were found in the ethanol (aq) extract and only 10 in the pH 1.78 acetic acid extract and 11 in the pH 2.17 acetic acid extract of the cane toad skins. Therefore, the aqueous and 60% ethanolic extracts present the greatest potential for ongoing development in our assays. There have been no previous reports on the identification of many of the constituents we have here identified in Australian cane toad skins. These findings, while somewhat consistent with findings in toad skins in other countries, identifies the presence of potential bioactive constituents. Our results showed that HPLC-ESI-Q-TOF-MS/MS is an effective method to characterise and identify components in Australian cane toad skin extracts. Chemical profiling is an essential initial step in the identification and therapeutic exploitation of bioactive agents present in Australian cane toad skin extracts.
Determination of coumarins in the roots of Angelica dahurica by supercritical fluid chromatography.Thursday, July 21, 2016
Pfeifer I, Murauer A, Ganzera M,
Journal of pharmaceutical and biomedical analysis. 11-Jul-2016
The fact that supercritical fluid chromatography (SFC) offers many desirable features is known for a long time. Yet, the number of applications on natural products is still limited, because robust and user-friendly instrumentation became available just a few years ago. As coumarins hardly have been studied by this technique we developed the first SFC assay for their determination in crude plant material. After method optimization eight standard compounds, including simple coumarins, linear and angular furanocoumarins, could be baseline separated in 6min using an Acquity UPC(2) CSH Fluoro-Phenyl 1.7μm column with supercritical CO2, methanol and diethylamine as mobile phase. Method validation confirmed that the assay is linear (R(2)≥0.9995), precise (intra-day variation≤5.8%; inter-day variation≤4.4%) and accurate (recovery rates from 96.5 to 104.2%). Detection limits determined at 300nm were below 2ng on-column, and the method showed to be well suited for the analysis of coumarins in Angelica dahurica roots. It was observed that qualitative as well as quantitative composition vary significantly. In all samples Imperatorin (0.09-0.28%) was the major coumarin, followed either by Isoimperatorin or Oxypeucedanin; the total coumarin content ranged from 0.16 to 0.77%. The results were in good agreement to published data, so that because of its speed and green nature SFC is definitely an interesting alternative for the analysis of this important class of natural products.
Peripheral blood microvesicles secretion is influenced by storage time, temperature, and anticoagulants.Friday, July 22, 2016
Wisgrill L, Lamm C, Hartmann J, Preißing F, Dragosits K, Bee A, Hell L, Thaler J, Ay C, Pabinger I, Berger A, Spittler A,
Cytometry. Part A : the journal of the International Society for Analytical Cytology. Jul-2016
Microvesicles (MVs) are small membrane bound vesicles released from various cell types after activation or apoptosis. In the last decades, MVs received an increased interest as biomarkers in inflammation, coagulation and cancer. However, standardized pre-analytical steps are crucial for the minimization of artifacts in the MV analysis. Thus, this study evaluated the MV release in whole blood samples under the influence of different anticoagulants, storage time and various temperature conditions. Samples were collected from healthy probands and processed immediately, after 4, 8, 24 and 48 hours at room temperature (RT) or 4°C. To identify MV subpopulations, platelet free plasma (PFP) was stained with Annexin V, calcein AM, CD15, CD41 and CD235a. Analysis was performend on a CytoFLEX flow cytometer. Procoagulatory function of MVs was measured using a phospholipid dependent activity and a tissue factor MVactivity assay. Without prior storage, sodium citrate showed the lowest MV count compared to heparin and EDTA. Interestingly, EDTA showed a significant release of myeloid-derived MVs (MMVs) compared to sodium citrate. Sodium citrate showed a stable MV count at RT in the first 8 hours after blood collection. Total MV counts increased after 24 hours in sodium citrated or heparinzed blood which was related to all subpopulations. Interestingly, EDTA showed stable platelet-derived MV (PMV) and erythrocyte-derived MV (EryMV) count at RT over a 48 h period. In addition, the procoagulatory potential increased significantly after 8-hour storage. Based on both, this work and literature data, the used anticoagulant, storage time and storage temperature differently influence the analysis of MVs within 8 hours. To date, sodium citrated tubes are recommended for MV enumeration and functional analysis. EDTA tubes might be an option for the clinical routine due to stable PMV and EryMV counts. These new approaches need to be validated in a clinical laboratory setting before being applied to patient studies. © 2016 International Society for Advancement of Cytometry.
Combined CD44, c-MET, and EGFR expression in p16-positive and p16-negative head and neck squamous cell carcinomas.Thursday, July 21, 2016
Baschnagel AM, Tonlaar N, Eskandari M, Kumar T, Williams L, Hanna A, Pruetz BL, Wilson GD,
Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology. 21-Jul-2016
High CD44 expression is associated with high c-MET expression, p16-negative tumors, and EGFR-positive tumors. The combination of these markers predicts for poor prognosis in HNSCC patients treated with chemoradiation.
The cardiovascular markers copeptin and high-sensitive C-reactive protein decrease following specific therapy for primary aldosteronism.Thursday, July 21, 2016
Remde H, Dietz A, Emeny R, Riester A, Peters A, de Las Heras Gala T, Then C, Seissler J, Beuschlein F, Reincke M, Quinkler M,
Journal of hypertension. 19-Jul-2016
Copeptin and hsCRP levels decrease following specific primary aldosteronism therapy reflecting successful cardiovascular risk reduction. However, they are no independent predictors regarding cure of primary aldosteronism.
A mitochondria-targeted antioxidant can inhibit peroxidase activity of cytochrome c by detachment of the protein from liposomes.Thursday, July 21, 2016
Firsov AM, Kotova EA, Orlov VN, Antonenko YN, Skulachev VP,
FEBS letters. 21-Jul-2016
Interaction of cytochrome c with cardiolipin converts this respiratory chain electron-transfer protein into a peroxidase, supposedly involved in mitochondria-mediated apoptosis initiation. Liposome membrane permeabilization provoked by peroxidase activity of the cytochrome c/cardiolipin complex has been previously shown to be suppressed by conventional antioxidants. Here, the mitochondria-targeted antioxidant SkQ1 (plastoquinonyl-decyl-triphenylphosphonium) was found to strongly inhibit both cytochrome c/cardiolipin peroxidase activity and the permeabilization of liposomes composed of phosphatidylcholine and cardiolipin. A number of binding assays revealed a significant inhibiting effect of SkQ1 on cytochrome c binding to liposomes, thus suggesting that SkQ1-mediated protection of liposomes from the cytochrome c/H2 O2 -induced permeabilization involved distortion of the cytochrome c-membrane binding. It is suggested that antioxidant and antiapoptotic effects of alkyltriphenylphosphonium cations can be related to prevention of cytochrome c/cardiolipin interaction. This article is protected by copyright. All rights reserved.
Radiation-induced functional connectivity alterations in nasopharyngeal carcinoma patients with radiotherapy.Friday, July 22, 2016
Ma Q, Wu D, Zeng LL, Shen H, Hu D, Qiu S,
Medicine. Jul-2016
The study aims to investigate the radiation-induced brain functional alterations in nasopharyngeal carcinoma (NPC) patients who received radiotherapy (RT) using functional magnetic resonance imaging (fMRI) and statistic scale.The fMRI data of 35 NPC patients with RT and 24 demographically matched untreated NPC patients were acquired. Montreal Cognitive Assessment (MoCA) was also measured to evaluate their global cognition performance. Multivariate pattern analysis was performed to find the significantly altered functional connections between these 2 groups, while the linear correlation level was detected between the altered functional connections and the MoCA scores.Forty-five notably altered functional connections were found, which were mainly located between 3 brain networks, the cerebellum, sensorimotor, and cingulo-opercular. With strictly false discovery rate correction, 5 altered functional connections were shown to have significant linear correlations with the MoCA scores, that is, the connections between the vermis and hippocampus, cerebellum lobule VI and dorsolateral prefrontal cortex, precuneus and dorsal frontal cortex, cuneus and middle occipital lobe, and insula and cuneus. Besides, the connectivity between the vermis and hippocampus was also significantly correlated with the attention score, 1 of the 7 subscores of the MoCA.The present study provides new insights into the radiation-induced functional connectivity impairments in NPC patients. The results showed that the RT may induce the cognitive impairments, especially the attention alterations. The 45 altered functional connections, especially the 5 altered functional connections that were significantly correlated to the MoCA scores, may serve as the potential biomarkers of the RT-induced brain functional impairments and provide valuable targets for further functional recovery treatment.
Hyperaldosteronism and cardiovascular risk in patients with autosomal dominant polycystic kidney disease.Friday, July 22, 2016
Lai S, Petramala L, Mastroluca D, Petraglia E, Di Gaeta A, Indino E, Panebianco V, Ciccariello M, Shahabadi HH, Galani A, Letizia C, D'Angelo AR,
Medicine. Jul-2016
Hypertension is commonly associated with autosomal dominant polycystic kidney disease (ADPKD), often discovered before the onset of renal failure, albeit the pathogenetic mechanisms are not well elucidated. Hyperaldosteronism in ADPKD may contribute to the development of insulin resistance and endothelial dysfunction, and progression of cardiorenal disease. The aim of study was to evaluate the prevalence of primary aldosteronism (PA) in ADPKD patients and identify some surrogate biomarkers of cardiovascular risk.We have enrolled 27 hypertensive ADPKD patients with estimated glomerular filtration rate (eGFR) ≥ 60 mL/min, evaluating the renin-angiotensin-aldosterone system (RAAS), inflammatory indexes, nutritional status, homocysteine (Hcy), homeostasis model assessment-insulin resistance (HOMA-IR), mineral metabolism, microalbuminuria, and surrogate markers of atherosclerosis [carotid intima media thickness (cIMT), ankle/brachial index (ABI), flow mediated dilation (FMD), renal resistive index (RRI) and left ventricular mass index (LVMI)]. Furthermore, we have carried out the morpho-functional magnetic resonance imaging (MRI) with high-field 3 T Magnetom Avanto.We have divided patients into group A, with normal plasma aldosterone concentration (PAC) and group B with PA, present in 9 (33%) of overall ADPKD patients. Respect to group A, group B showed a significant higher mean value of LVMI, HOMA-IR and Hcy (P = 0.001, P = 0.004, P = 0.018; respectively), and a lower value of FMD and 25-hydroxyvitamin D (25-OH-VitD) (P = 0.037, P = 0.019; respectively) with a higher prevalence of non-dipper pattern at Ambulatory Blood Pressure Monitoring (ABPM) (65% vs 40%, P < 0.05) at an early stage of the disease.In this study, we showed a high prevalence of PA in ADPKD patients, associated to higher LVMI, HOMA-IR, Hcy, lower FMD, and 25-OH-VitD, considered as surrogate markers of atherosclerosis, compared to ADPKD patients with normal PAC values. Our results indicate a higher overall cardiovascular risk in ADPKD patients with inappropriate aldosterone secretion, and a screening for PA in all patients with ADPKD is recommended.
Microfluidic Immuno-Biochip for Detection of Breast Cancer Biomarkers Using Hierarchical Composite of Porous Graphene and Titanium Dioxide Nanofibers.Thursday, July 21, 2016
Ali MA, Mondal K, Jiao Y, Oren S, Xu Z, Sharma A, Dong L,
ACS applied materials & interfaces. 21-Jul-2016
We report on a label-free microfluidic immunosensor with femtomolar sensitivity and high selectivity for early detection of epidermal growth factor receptor 2 (EGFR2 or ErbB2) proteins. This sensor utilizes a uniquely structured immunoelectrode made of porous hierarchical graphene foam (GF) modified with electrospun carbon-doped titanium dioxide nanofibers (nTiO2) as an electrochemical working electrode. Due to excellent biocompatibility, intrinsic surface defects, high reaction kinetics, and good stability for proteins, anatase nTiO2 are ideal for electrochemical sensor applications. The three-dimensional and porous features of GF allow nTiO2 to penetrate and attach to the surface of the GF by physical adsorption. Combining GF with functional nTiO2 yields high charge transfer resistance, large surface area, and porous access to the sensing surface by the analyte, resulting in new possibilities for the development of electrochemical immunosensors. Here, the enabling of EDC-NHS chemistry covalently immobilized the antibody of ErbB2 (anti-ErbB2) on the GF-nTiO2 composite. To obtain a compact sensor architecture, the composite working electrode was designed to hang above the gold counter electrode in a microfluidic channel. The sensor underwent differential pulse voltammetry and electrochemical impedance spectroscopy to quantify breast cancer biomarkers. The two methods had high sensitivities of 0.585 µA µM-1 cm-2 and 43.7 kΩ µM-1 cm-2 in a wide concentration range of target ErbB2 antigen from 1 × 10-15 to 0.1 × 10-6 M, and from 1 × 10-13 to 0.1 × 10-6, respectively. Utilization of the specific recognition element (i.e., anti-ErbB2) results in high specificity, even in the presence of identical members of the EGFR family of receptor tyrosine kinases, such as ErbB3 and ErbB4. Many promising applications in the field of electrochemical detection of chemical and biological species will derive from the integration of the porous GF-nTiO2 composite into microfluidic devices.
Mining enzyme diversity of transcriptome libraries through DNA synthesis for benzylisoquinoline alkaloid pathway optimization in yeast.Thursday, July 21, 2016
Narcross L, Bourgeois L, Fossati E, Burton E, Martin VJ,
ACS synthetic biology. 21-Jul-2016
The ever-increasing quantity of data deposited to GenBank is a valuable resource for mining new enzyme activities. Falling costs of DNA synthesis enables metabolic engineers to take advantage of this resource for identifying superior or novel enzymes for pathway optimization. Previously, we reported synthesis of the benzylisoquinoline alkaloid dihydrosanguinarine in yeast from norlaudanosoline at a molar conversion of 1.5%. Molar conversion could be improved by reduction of the side-product N-methylcheilanthifoline, a key bottleneck in dihydrosanguinarine biosynthesis. Two pathway enzymes, an N-methyltransferase and a cytochrome P450 of the CYP719A subfamily, were implicated in the synthesis of the side-product. Here, we conducted an extensive screen to identify enzyme homologs whose co-expression reduces side-product synthesis. Phylogenetic trees were generated from multiple sources of sequence data to identify a library of candidate enzymes that were purchased codon-optimized and pre-cloned into expression vectors designed to facilitate high-throughput analysis of gene expression as well as activity assay. Simple in vivo assays were sufficient to guide the selection of superior enzyme homologs that ablated the synthesis of the side-product, and improved molar conversion of norlaudanosoline to dihydrosanguinarine to 10%.
A Fluorescent Transport Assay Enables Studying AmpG Permeases Involved in Peptidoglycan Recycling and Antibiotic Resistance.Thursday, July 21, 2016
Perley-Robertson GE, Yadav AK, Winogrodzki JL, Stubbs KA, Mark BL, Vocadlo DJ,
ACS chemical biology. 21-Jul-2016
Inducible AmpC β-lactamases deactivate a broad-spectrum of β-lactam antibiotics and afford antibiotic resistance in many Gram-negative bacteria. The disturbance of peptidoglycan recycling caused by β-lactam antibiotics leads to accumulation of GlcNAc-1,6-anhydroMurNAc-peptides, which are transported by AmpG to the cytoplasm where they are processed into AmpC inducers. AmpG transporters are poorly understood, however, their loss restores susceptibility toward β-lactam antibiotics, highlighting AmpG as a potential target for resistance-attenuating therapeutics. We prepare a GlcNAc-1,6-anhydroMurNAc-fluorophore conjugate and, using live E. coli spheroplasts, quantitatively analyze its transport by AmpG and inhibition of this process by a competing substrate. Further, we use this transport assay to evaluate the function of two AmpG homologues from Pseudomonas aeruginosa and show that P. aeruginosa AmpG (Pa-AmpG) but not AmpP (Pa-AmpP) transports this probe substrate. We corroborate these results by AmpC induction assays with Pa-AmpG and Pa-AmpP. This fluorescent AmpG probe and spheroplast-based transport assay will enable improved understanding of PG recycling and of permeases from the major facilitator superfamily of transport proteins, and may aid in identification of AmpG antagonists that combat AmpC-mediated resistance toward β-lactam antibiotics.
Development and validation of high-resolution melting (HRM) markers derived from Rysto STS markers for high-throughput marker-assisted selection of potato carrying Rysto.Thursday, July 21, 2016
Nie X, Sutherland D, Dickison V, Singh M, Murphy AM, De Koeyer D,
Phytopathology. 20-Jul-2016
Sequence analysis of the chromosome region harbouring the STS markers YES3-3A and YES3-3B for Rysto, a gene responsible for extreme resistance to Potato virus Y (PVY) in potato, was performed in tetraploid potato 'Barbara' (Rrrr) and 'AC Chaleur' (rrrr) as well as their progeny selections. Three and two sequence variants were identified in Barbara/resistant (R) selections and AC Chaleur/susceptible (S) selections, respectively. Further analysis indicates that the variant with a 21-nucleotide (nt) deletion is likely the chromosome copy harbouring the STS markers. Two primer pairs, one targeting the region containing the 21-nt deletion and the other targeting the region anchoring the YES3-3A reverse primer, were designed. As anticipated, pair one produced two visible fragments in Barbara/R bulk and one visible fragment in AC Chaleur/S bulk; pair two produced one visible fragment in all samples. When subjected to high-resolution melting (HRM) analysis, two distinct melting profiles for R and S samples were observed. Analysis of 147 progeny of Barbara × AC Chaleur revealed 72 and 75 progeny with R and S melting profiles, respectively, which was consistent with YES3-3A and YES3-3B assays and phenotyping analysis, thus demonstrating the potential of HRM profiles as novel molecular markers for Rysto. The efficacy of the newly developed HRM markers for high-throughput marker-assisted selection for Rysto-conferred resistance to PVY was validated further with three populations involving Barbara as the resistant parent.
PCR-mediated detection and quantification of the Goss's Wilt pathogen Clavibacter michiganensis subsp. nebraskensis via a novel gene target.Thursday, July 21, 2016
McNally RR, Ishimaru C, Malvick D,
Phytopathology. 20-Jul-2016
Goss's leaf blight and wilt of maize (corn) is a significant and reemerging disease caused by the bacterium Clavibacter michiganensis subsp. nebraskensis (Cmn). Despite its importance, molecular tools for diagnosing and studying this disease remain limited. We report the identification of CMN_01184 as a novel gene target and its use in conventional PCR (cPCR) and SYBR green-based quantitative PCR (qPCR) assays for specific detection and quantification of Cmn. The cPCR and qPCR assays based on primers targeting CMN_01184 specifically amplified only Cmn among a diverse collection of 129 bacterial and fungal isolates, including multiple maize bacterial and fungal pathogens, environmental organisms from agricultural fields, and all known subspecies of C. michiganensis. Specificity of the assays for detection of only Cmn was also validated with field samples of Cmn-infected and uninfected maize leaves and Cmn-infested and uninfested soil. Detection limits were determined at 30 and 3 ng of pure Cmn DNA, and 100 and 10 CFU of Cmn for the cPCR and qPCR assays, respectively. Infection of maize leaves by Cmn was quantified from infected field samples and standardized using an internal maize DNA control. These novel, specific, and sensitive PCR assays based on CMN_01184 are effective for diagnosis of Goss's wilt and studies of the epidemiology and host-pathogen interactions of Cmn.
Artificial MicroRNAs as Novel Secreted Reporters for Cell Monitoring in Living Subjects.Friday, July 22, 2016
Ronald JA, D'Souza AL, Chuang HY, Gambhir SS,
PloS one. 2016
Reporter genes are powerful technologies that can be used to directly inform on the fate of transplanted cells in living subjects. Imaging reporter genes are often employed to quantify cell number, location(s), and viability with various imaging modalities. To complement this, reporters that are secreted from cells can provide a low-cost, in vitro diagnostic test to monitor overall cell viability at relatively high frequency without knowing the locations of all cells. Whereas protein-based secretable reporters have been developed, an RNA-based reporter detectable with amplification inherent PCR-based assays has not been previously described. MicroRNAs (miRNAs) are short non-coding RNAs (18-22 nt) that regulate mRNA translation and are being explored as relatively stable blood-based disease biomarkers. We developed an artificial miRNA-based secreted reporter, called Sec-miR, utilizing a coding sequence that is not expressed endogenously and does not have any known vertebrate target. Sec-miR was detectable in both the cells and culture media of transiently transfected cells. Cells stably expressing Sec-miR also reliably secreted it into the culture media. Mice implanted with parental HeLa cells or HeLa cells expressing both Sec-miR and the bioluminescence imaging (BLI) reporter gene Firefly luciferase (FLuc) were monitored over time for tumor volume, FLuc signal via BLI, and blood levels of Sec-miR. Significantly (p<0.05) higher Sec-miR was found in the blood of mice bearing Sec-miR-expressing tumors compared to parental cell tumors at 21 and 28 days after implantation. Importantly, blood Sec-miR reporter levels after day 21 showed a trend towards correlation with tumor volume (R2 = 0.6090; p = 0.0671) and significantly correlated with FLuc signal (R2 = 0.7067; p<0.05). Finally, we could significantly (p<0.01) amplify Sec-miR secretion into the cell media by chaining together multiple Sec-miR copies (4 instead of 1 or 2) within an expression cassette. Overall, we show that a novel complement of BLI together with a unique Sec-miR reporter adds an in vitro RNA-based diagnostic to enhance the monitoring of transplanted cells. While Sec-miR was not as sensitive as BLI for monitoring cell number, it may be more sensitive than clinically-relevant positron emission tomography (PET) reporter assays. Future work will focus on improving cell detectability via improved secretion of Sec-miR reporters from cells and more sensitive detection platforms, as well as, exploring other miRNA sequences to allow multiplexed monitoring of more than one cell population at a time. Continued development may lead to more refined and precise monitoring of cell-based therapies.
Establishment of a Predictive In Vitro Assay for Assessment of the Hepatotoxic Potential of Oligonucleotide Drugs.Friday, July 22, 2016
Sewing S, Boess F, Moisan A, Bertinetti-Lapatki C, Minz T, Hedtjaern M, Tessier Y, Schuler F, Singer T, Roth AB,
PloS one. 2016
Single stranded oligonucleotides (SSO) represent a novel therapeutic modality that opens new space to address previously undruggable targets. In spite of their proven efficacy, the development of promising SSO drug candidates has been limited by reported cases of SSO-associated hepatotoxicity. The mechanisms of SSO induced liver toxicity are poorly understood, and up to now no preclinical in vitro model has been established that allows prediction of the hepatotoxicity risk of a given SSO. Therefore, preclinical assessment of hepatic liability currently relies on rodent studies that require large cohorts of animals and lengthy protocols. Here, we describe the establishment and validation of an in vitro assay using primary hepatocytes that recapitulates the hepatotoxic profile of SSOs previously observed in rodents. In vitro cytotoxicity upon unassisted delivery was measured as an increase in extracellular lactate dehydrogenase (LDH) levels and concomitant reduction in intracellular glutathione and ATP levels after 3 days of treatment. Furthermore, toxic, but not safe, SSOs led to an increase in miR-122 in cell culture supernatants after 2 days of exposure, revealing the potential use of miR122 as a selective translational biomarker for detection of SSO-induced hepatotoxicity. Overall, we have developed and validated for the first time a robust in vitro screening assay for SSO liver safety profiling which allows rapid prioritization of candidate molecules early on in development.
Storage of Erythrocytes Induces Suicidal Erythrocyte Death.Thursday, July 21, 2016
Lang E, Pozdeev VI, Xu HC, Shinde PV, Behnke K, Hamdam JM, Lehnert E, Scharf RE, Lang F, Häussinger D, Lang KS, Lang PA,
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology. 21-Jul-2016
Storage of RBC triggers eryptosis by Ca2+ and erythropoietin sensitive mechanisms.
Notes from the Field: Rickettsia parkeri Rickettsiosis - Georgia, 2012-2014.Friday, July 22, 2016
Straily A, Feldpausch A, Ulbrich C, Schell K, Casillas S, Zaki SR, Denison AM, Condit M, Gabel J, Paddock CD,
MMWR. Morbidity and mortality weekly report. 2016
During 2012-2014, five cases of Rickettsia parkeri rickettsiosis were identified by a single urgent care practice in Georgia, located approximately 40 miles southwest of Atlanta. Symptom onset occurred during June-October, and all patients had a known tick bite. Patients ranged in age from 27 to 72 years (median = 53 years), and all were male. The most commonly reported initial signs were erythema (n = 3) and swelling (n = 2) at the site of the bite. Two patients reported fever and a third patient reported a rash and lymphadenopathy without fever. Other symptoms included myalgia (n = 3), chills (n = 3), fatigue (n = 2), arthralgia (n = 2), and headache (n = 2). Eschar biopsy specimens were collected from each patient using a 4-mm or 5-mm punch and placed in 10% neutral buffered formalin or sterile saline. These specimens were tested by immunohistochemical (IHC) stains, quantitative polymerase chain reaction (qPCR) assays, or cell culture isolation to determine if there was evidence of infection with a Rickettsia species (1). IHC evidence of spotted fever group rickettsiae was found in the eschar biopsy specimens in all five cases. In four cases, the biopsy specimens were also positive for R. parkeri by qPCR. The fifth case (specimen positive only by IHC testing) was considered a probable R. parkeri case based on clinical signs and symptoms. R. parkeri was grown in cell culture from one specimen from which isolation was attempted. All patients were treated with oral doxycycline (100 mg twice daily) for a minimum of 10 days, and all recovered.
Matrix-assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) Can Precisely Discriminate the Lineages of Listeria monocytogenes and Species of Listeria.Friday, July 22, 2016
Ojima-Kato T, Yamamoto N, Takahashi H, Tamura H,
PloS one. 2016
The genetic lineages of Listeria monocytogenes and other species of the genus Listeria are correlated with pathogenesis in humans. Although matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has become a prevailing tool for rapid and reliable microbial identification, the precise discrimination of Listeria species and lineages remains a crucial issue in clinical settings and for food safety. In this study, we constructed an accurate and reliable MS database to discriminate the lineages of L. monocytogenes and the species of Listeria (L. monocytogenes, L. innocua, L. welshimeri, L. seeligeri, L. ivanovii, L. grayi, and L. rocourtiae) based on the S10-spc-alpha operon gene encoded ribosomal protein mass spectrum (S10-GERMS) proteotyping method, which relies on both genetic information (genomics) and observed MS peaks in MALDI-TOF MS (proteomics). The specific set of eight biomarkers (ribosomal proteins L24, L6, L18, L15, S11, S9, L31 type B, and S16) yielded characteristic MS patterns for the lineages of L. monocytogenes and the different species of Listeria, and led to the construction of a MS database that was successful in discriminating between these organisms in MALDI-TOF MS fingerprinting analysis followed by advanced proteotyping software Strain Solution analysis. We also confirmed the constructed database on the proteotyping software Strain Solution by using 23 Listeria strains collected from natural sources.
Probing the Rare Biosphere of the North-West Mediterranean Sea: An Experiment with High Sequencing Effort.Friday, July 22, 2016
Crespo BG, Wallhead PJ, Logares R, Pedrós-Alió C,
PloS one. 2016
High-throughput sequencing (HTS) techniques have suggested the existence of a wealth of species with very low relative abundance: the rare biosphere. We attempted to exhaustively map this rare biosphere in two water samples by performing an exceptionally deep pyrosequencing analysis (~500,000 final reads per sample). Species data were derived by a 97% identity criterion and various parametric distributions were fitted to the observed counts. Using the best-fitting Sichel distribution we estimate a total species richness of 1,568-1,669 (95% Credible Interval) and 5,027-5,196 for surface and deep water samples respectively, implying that 84-89% of the total richness in those two samples was sequenced, and we predict that a quadrupling of the present sequencing effort would suffice to observe 90% of the total richness in both samples. Comparing the HTS results with a culturing approach we found that most of the cultured taxa were not obtained by HTS, despite the high sequencing effort. Culturing therefore remains a useful tool for uncovering marine bacterial diversity, in addition to its other uses for studying the ecology of marine bacteria.
Biosimilar medical products - licensing, pharmacovigilance and interchangeability.Friday, July 22, 2016
Grozdanova A, Netkovska KA, Sterjev Z, Naumovska Z, Zarevski R, Dimovski A, Suturkova L,
Prilozi (Makedonska akademija na naukite i umetnostite. Oddelenie za medicinski nauki). 2016
The use of biological medicine has significantly increased in recent decades and has made substantial contributions to improving the effectiveness of therapies in many diseases. The expiration of patents of biological innovative medicines enables copies of those drugs called similar biological products (biosimilars) to be approved by regulatory authorities and to enter in clinical use. Biosimilars are comparable but not identical and are not a generic version of the innovator biological product. Although biosimilars undergo rigorous characterization as well as clinical studies to prove their safety and effectiveness, specific regulatory requirements for registration apply in the case of biosimilars. They are highly complex molecules and small changes in the production process can have major implications in its safety and effectiveness profile. The availability of biosimilars enhances competition, with the potential to improve patient access to biological medicines and to contribute to the financial sustainability of healthcare systems. In order to be certain that a biosimilar reaches its potential in clinical use, an intensive pharmacovigilance monitoring system must be established in order to prove the true similarity between the original biologic and its biosimilar. There is a need for further guidance and resolution of the ongoing discussions on biosimilar labelling, naming, pharmacovigilance and substitution in order to ensure effective and appropriate use of biosimilars in clinical practice.
MUC1 immunotherapy against a metastatic mammary adenocarcinoma model: Importance of IFN-gamma.Friday, July 22, 2016
Lees CJ, Smorodinsky N, Horn G, Wreschner DH, McKenzie IF, Pietersz G, Stojanovska L, Apostolopoulos V,
Prilozi (Makedonska akademija na naukite i umetnostite. Oddelenie za medicinski nauki). 2016
Immunotherapy using mucin 1 (MUC1) linked to oxidised mannan (MFP) was investigated in an aggressive MUC1+ metastatic tumour, DA3-MUC1 because, unlike many MUC1+ tumour models, DA3-MUC1 is not spontaneously rejected in mice making it an alternative model for immunotherapy studies. Further, DA3-MUC1 cells are resistant to lysis by anti-MUC1 cytotoxic T cells (CTLs). The inability of DA3-MUC1 tumours to be rejected in naïve mice as well as vaccination to MUC1 was attributed to a deficiency of expression of MHC class I molecules on the tumour cell surface. In vitro and in vivo analysis of subcutaneous tumours and lung metastases demonstrated that DA3-MUC1 tumour cells have a low expression (< 6%) of MHC class I which can be upregulated (> 90%) following culturing with IFN-γ. Results from flow cytometry analysis and immunoperoxidase staining indicated that the in vitro up-regulation of MHC class I could be maintained for up to seven days in vivo, without affecting the expression levels of MUC1 antigen. Interestingly, MUC1-specific CTL that lyse DA3-MUC1 targets in vitro were induced in MFP immunised mice but failed to protect mice from a DA3-MUC1 tumour challenge. These results highlight the importance of MHC class I molecules in the induction of anti-tumour immunity and the MFP immune response.
4th Rare Disease South Eastern Europe (See) Meeting Skopje, Macedonia (November 14th, 2015).Friday, July 22, 2016
Gucev Z, Tasic V, Polenakovic M,
Prilozi (Makedonska akademija na naukite i umetnostite. Oddelenie za medicinski nauki). 2015
The 4th meeting on rare diseases in South Eastern Europe (SEE) was held in Skopje, at the Macedonian Academy of Sciences and Arts (MASA) on the 14th of November 2015. The focuses were metabolic, rare brain diseases as well as the rare dysmorphic syndrome. The authors of the report are particularly keen on stating that one of the main goals of the meeting, namely to help the treatment of patients with rare disease has begun to bear fruits. The talk on an iminosugar-based pharmacological chaperone compound as a drug candidate for the treatment of GM1-gangliosidosis and mucopolysaccharidosis IVB (Morquio disease type B) was enlightening. To date, there is no treatment available to be offered to patients, but chaperones lead mutated proteins to adopt a native-like conformation and to successfully traffic to their normal cellular destination. DORPHAN is developing an iminosugar-based pharmacological chaperone compound for the treatment of GM1-gangliosidosis and mucopolysaccharidosis IVB. A talk on recent developments in the laboratory diagnosis of mucopolysaccharidoses (MPS) was particularly interesting, covering the laboratory diagnosis of the MPS diseases by a strategy of clinical examination, biochemical analysis of urine samples, enzyme tests and genetic characterization of underlying mutations. New techniques were developed, including analysis of urinary glycosaminoglycans with tandem mass spectrometry, miniaturized enzyme tests or novel synthetic substrates for enzyme assays using mass spectrometry detection of products using dried blood spots. Feasibility and cost-effectiveness of these methods in newborn screening programs have been demonstrated. Neuromuscular RDs, and especially familial amyloid polyneuropathy (FAP) were a topic of the Bulgarian colleagues. Diagnosis, screening and the role of microglia were also topics of particular interest. In summary, this year RD meeting was exciting and productive on a wide range of diseases and on a novel insights on diagnosis and treatment. New methods are expanding our capabilities for a fast and precise diagnosis. Novel knowledge offers better distinction on whom to treat with which medications (e.g. steroid dependent nephrotic syndrome). Novel diseases or variants are published (segmental overgrowth). The authors of the report are particularly keen on stating that one of the main goals of the meeting, namely to help the treatment of patients with rare disease has begun to bear fruits. Namely, the Health Fund of Macedonia for the first time treats the patients with Gaucher's disease. We are hopeful that the number of patients treated for Gaucher's disease and the number of treated patients with other treatable RDs diseases will continue to grow.
S100B raises the alert in subarachnoid hemorrhage.Thursday, July 21, 2016
Chong ZZ,
Reviews in the neurosciences. 21-Jul-2016
Subarachnoid hemorrhage (SAH) is a devastating disease with high mortality and mobility, the novel therapeutic strategies of which are essentially required. The calcium binding protein S100B has emerged as a brain injury biomarker that is implicated in pathogenic process of SAH. S100B is mainly expressed in astrocytes of the central nervous system and functions through initiating intracellular signaling or via interacting with cell surface receptor, such as the receptor of advanced glycation end products. The biological roles of S100B in neurons have been closely associated with its concentrations, resulting in either neuroprotection or neurotoxicity. The levels of S100B in the blood have been suggested as a biomarker to predict the progress or the prognosis of SAH. The role of S100B in the development of cerebral vasospasm and brain damage may result from the induction of oxidative stress and neuroinflammation after SAH. To get further insight into mechanisms underlying the role of S100B in SAH based on this review might help us to find novel therapeutic targets for SAH.
Mechanistic Investigation of a Non-Heme Iron Enzyme Catalyzed Epoxidation in (-)-4'-Methoxycyclopenin Biosynthesis.Thursday, July 21, 2016
Chang WC, Li J, Lee JL, Cronican AA, Guo Y,
Journal of the American Chemical Society. 21-Jul-2016
Mechanisms have been proposed for α-KG dependent non-heme iron enzyme catalyzed oxygen atom insertion into an olefinic moiety in various natural products, but not examined in detail. Using a combination of methods including transient kinetics, Mössbauer spectroscopy and mass spectrometry, we demonstrate that AsqJ catalyzed (-)-4'methoxy-cyclopenin formation uses a high-spin Fe(IV)-oxo intermediate to carry out epoxidation. Furthermore, product analysis on 16O/18O isotope incorporation from the reactions using the native substrate, 4'-methoxy-dehydrocyclopeptin, and a mechanistic probe, dehydrocyclopeptin, reveals evidence supporting oxo-hydroxo tautomerism of the Fe(IV)-oxo species in the non-heme iron enzyme catalysis.
Circulating Blood Monocyte Subclasses and Lipid-Laden Adipose Tissue Macrophages in Human Obesity.Friday, July 22, 2016
Pecht T, Haim Y, Bashan N, Shapiro H, Harman-Boehm I, Kirshtein B, Clément K, Shai I, Rudich A,
PloS one. 2016
Collectively, although circulating blood NCM are unlikely direct functional precursor cells for adipose tissue foam cells, their increased percentage in the circulation may clinically reflect higher lipid content in visceral ATMs.
Enhanced Eryptosis Following Exposure to Dolutegravir.Thursday, July 21, 2016
Al Mamun Bhuyan A, Signoretto E, Bissinger R, Lang F,
Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology. 21-Jul-2016
Dolutegravir triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part due to Ca2+ entry, ceramide formation and oxidative stress.
11-Ketotestosterone and 11-Ketodihydrotestosterone in Castration Resistant Prostate Cancer: Potent Androgens Which Can No Longer Be Ignored.Friday, July 22, 2016
Pretorius E, Africander DJ, Vlok M, Perkins MS, Quanson J, Storbeck KH,
PloS one. 2016
Dihydrotestosterone (DHT) is regarded as the most potent natural androgen and is implicated in the development and progression of castration resistant prostate cancer (CRPC). Under castrate conditions, DHT is produced from the metabolism of the adrenal androgen precursors, DHEA and androstenedione. Recent studies have shown that the adrenal steroid 11β-hydroxyandrostenedione (11OHA4) serves as the precursor to the androgens 11-ketotestosterone (11KT) and 11-ketodihydrotestosterone (11KDHT). In this study we comprehensively assess the androgenic activity of 11KT and 11KDHT. This is the first study, to our knowledge, to show that 11KT and 11KDHT, like T and DHT, are potent and efficacious agonists of the human androgen receptor (AR) and induced both the expression of representative AR-regulated genes as well as cellular proliferation in the androgen dependent prostate cancer cell lines, LNCaP and VCaP. Proteomic analysis revealed that 11KDHT regulated the expression of more AR-regulated proteins than DHT in VCaP cells, while in vitro conversion assays showed that 11KT and 11KDHT are metabolized at a significantly lower rate in both LNCaP and VCaP cells when compared to T and DHT, respectively. Our findings show that 11KT and 11KDHT are bona fide androgens capable of inducing androgen-dependant gene expression and cell growth, and that these steroids have the potential to remain active longer than T and DHT due to the decreased rate at which they are metabolised. Collectively, our data demonstrates that 11KT and 11KDHT likely play a vital, but overlooked, role in the development and progression of CRPC.
First Case of Legionnaire's Disease Caused by Legionella anisa in Spain and the Limitations on the Diagnosis of Legionella non-pneumophila Infections.Friday, July 22, 2016
Vaccaro L, Izquierdo F, Magnet A, Hurtado C, Salinas MA, Gomes TS, Angulo S, Salso S, Pelaez J, Tejeda MI, Alhambra A, Gómez C, Enríquez A, Estirado E, Fenoy S, Del Aguila C,
PloS one. 2016
Legionnaires' disease is a severe form of pneumonia, with worldwide relevance, caused by Legionella spp. Approximately 90% of all cases of legionellosis are caused by Legionella pneumophila, but other species can also be responsible for this infection. These bacteria are transmitted by inhalation of aerosols or aspiration of contaminated water. In Spain, environmental studies have demonstrated the presence of Legionella non-pneumophila species in drinking water treatment plants and water distribution networks. Aware that this evidence indicates a risk factor and the lack of routine assays designed to detect simultaneously diverse Legionella species, we analyzed 210 urine samples from patients presenting clinical manifestations of pneumonia using a semi-nested PCR for partial amplification of the 16S rDNA gene of Legionella and a diagnostic method used in hospitals for Legionella antigen detection. In this study, we detected a total of 15 cases of legionellosis (7.1%) and the first case of Legionnaires' disease caused by L. anisa in Spain. While the conventional method used in hospitals could only detect four cases (1.9%) produced by L. pneumophila serogroup 1, using PCR, the following species were identified: Legionella spp. (10/15), L. pneumophila (4/15) and L. anisa (1/15). These results suggest the need to change hospital diagnostic strategies regarding the identification of Legionella species associated with this disease. Therefore, the detection of Legionella DNA by PCR in urine samples seems to be a suitable alternative method for a sensitive, accurate and rapid diagnosis of Legionella pneumonia, caused by L. pneumophila and also for L. non-pneumophila species.
Predictive models of cytotoxicity as mediated by exposure to chemicals or drugs.Friday, July 22, 2016
Moon H, Cong M,
SAR and QSAR in environmental research. Jun-2016
Predicting cytotoxicity is a challenging task because of the complex biological mechanisms behind it. Cytotoxicity due to toxin - biologically produced poison - is known to play a substantial role in a disease process. Two objectives in this research are to derive robust general predictive cytotoxicity models to minimize unnecessary toxicity. The first objective is to build accurate predictive statistical models for cytotoxicity data based on lymphoblastoid cell lines obtained from in vitro studies. This could be an important step for accomplishing a goal in biomedecial/biophamarceutical research, by obtaining the best medical outcomes by minimizing toxicity in regard to a person's genetic profile. The second objective is to build predictive models to predict population-level cytotoxicity for unknown compounds based on chemical structural features. These two objectives were accomplished by a proposed variable selection process, the random forests, and the least absolute shrinkage and selection operator method. We achieved an excellent prediction result with the random forests algorithm using SNP markers from the proposed approach, having the smallest root mean squared error among the teams which participated in the DREAM Toxicogenetics Challenge. Since chemical compounds for drugs have great influence on human health, the predictive statistical models for these objectives could be helpful to government agencies in relevant decision-making.
In-situ second harmonic generation by cancer cell targeting ZnO nanocrystals to effect photodynamic action in subcellular space.Thursday, July 21, 2016
Gu B, Pliss A, Kuzmin AN, Baev A, Ohulchanskyy TY, Damasco JA, Yong KT, Wen S, Prasad PN,
Biomaterials. 12-Jul-2016
This paper introduces the concept of in-situ upconversion of deep penetrating near infrared light via second harmonic generation from ZnO nanocrystals delivered into cells to effect photo activated therapies, such as photodynamic therapy, which usually require activation by visible light with limited penetration through biological tissues. We demonstrated this concept by subcellular activation of a photodynamic therapy drug, Chlorin e6, excited within its strong absorption Soret band by the second harmonic (SH) light, generated at 409 nm by ZnO nanocrystals, which were targeted to cancer cells and internalized through the folate-receptor mediated endocytosis. By a combination of theoretical modeling and experimental measurements, we show that SH light, generated in-situ by ZnO nanocrystals significantly contributes to activation of photosensitizer, leading to cell death through both apoptotic and necrotic pathways initiated in the cytoplasm. This targeted photodynamic action was studied using label-free Coherent Anti-Stokes Raman Scattering imaging of the treated cells to monitor changes in the distribution of native cellular proteins and lipids. We found that initiation of photodynamic therapy with upconverted light led to global reduction in the intracellular concentration of macromolecules, likely due to suppression of proteins and lipids synthesis, which could be considered as a real-time indicator of cellular damage from photodynamic treatment. In prospective applications this in-situ photon upconversion could be further extended using ZnO nanocrystals surface functionalized with a specific organelle targeting group, provided a powerful approach to identify and consequently maximize a cellular response to phototherapy, selectively initiated in a specific cellular organelle.
Fructose-coated nanodiamonds: promising platforms for treatment of human breast cancer.Thursday, July 21, 2016
Zhao J, Lai H, Lu H, Barner-Kowollik C, Stenzel MH, Xiao P,
Biomacromolecules. 21-Jul-2016
Well-defined carboxyl end-functionalized glycopolymer poly(1-O-methacryloyl-2,3:4,5-di-O-isopropylidene-β-D-fructopyranose) has been prepared via reversible addition-fragmentation chain transfer polymerization and grafted onto the surface of amine-functionalized nanodiamonds via a simple conjugation reaction. The properties of the nanodiamond-polymer hybrid materials are investigated using infrared spectroscopy, thermogravimetric analysis, dynamic light scattering, and transmission electron microscopy. The dispersibility of the nanodiamonds in aqueous solutions is significantly improved after the grafting of the glycopolymer. More interestingly, the cytotoxicity of amine-functionalized nanodiamonds is significantly decreased after decoration with the glycopolymer even at a high concentration (125 μg/mL). The nanodiamonds were loaded with doxorubicin to create a bioactive drug delivery carrier. The release of doxorubicin was faster in media of pH 5 than that in pH 7.4. The nanodiamond drug delivery systems with doxorubicin are used to treat breast cancer cells in 2D and 3D models. Although the 2D cell culture results indicate that all nanodiamonds-doxorubicin complexes are significantly less toxic than free doxorubicin, the glycopolymer-coated nanodiamonds-doxorubicin show higher cytotoxicity than free doxorubicin in the 3D spheroids after treatment for 8 days. The enhanced cytotoxicity of Poly(1-O-MAFru)62-ND-Dox in 3D spheroids may result from the sustained drug release and deep penetration of these nanocarriers, which play a role as "Trojan Horse". The massive cell death after eight-day incubation with Poly(1-O-MAFru)62-ND-Dox demonstrates that glycopolymer-coated nanodiamonds can be promising platforms for breast cancer therapy.
Gastroenterological endpoints in drug trials for cystic fibrosis.Thursday, July 21, 2016
Bodewes FA, Verkade HJ, Wilschanski M,
Pediatric pulmonology. 21-Jul-2016
The phenotype of cystic fibrosis includes a wide variety of clinical and biochemical gastrointestinal presentations. These gastrointestinal characteristics of the disease have come under renewed interest as potential outcome measures and clinical endpoints for therapeutic trials in cystic fibrosis. Established gastrointestinal clinical endpoints, like e.g. fecal elastase-1, are already used in trials. Other potential gastrointestinal outcome measures gather more scientific interest for evaluation in future trials. Gastrointestinal outcome measures look particularly relevant and promising for trials in CF patients with normal lung function or therapeutic studies in young children and infants. We review, the currently reported gastrointestinal effects of CFTR modulation therapies and discuss the potential of gastrointestinal outcome measures for therapeutic trials in cystic fibrosis. Pediatr Pulmonol.. © 2016 Wiley Periodicals, Inc.
Targeting of a Transporter to the Outer Apicoplast Membrane in the Human Malaria Parasite Plasmodium falciparum.Friday, July 22, 2016
Lim L, Sayers CP, Goodman CD, McFadden GI,
PloS one. 2016
Apicoplasts are vestigial plastids in apicomplexan parasites like Plasmodium, the causative agent of malaria. Apicomplexan parasites are dependant on their apicoplasts for synthesis of various molecules that they are unable to scavenge in sufficient quantity from their host, which makes apicoplasts attractive drug targets. Proteins known as plastid phosphate translocators (pPTs) are embedded in the outer apicoplast membrane and are responsible for the import of carbon, energy and reducing power to drive anabolic synthesis in the organelle. We investigated how a pPT is targeted into the outer apicoplast membrane of the human malaria parasite P. falciparum. We showed that a transmembrane domain is likely to act as a recessed signal anchor to direct the protein into the endomembrane system, and that a tyrosine in the cytosolic N-terminus of the protein is essential for targeting, but one or more, as yet unidentified, factors are also essential to direct the protein into the outer apicoplast membrane.
Host-Pathogen Coevolution and the Emergence of Broadly Neutralizing Antibodies in Chronic Infections.Friday, July 22, 2016
Nourmohammad A, Otwinowski J, Plotkin JB,
PLoS genetics. Jul-2016
The vertebrate adaptive immune system provides a flexible and diverse set of molecules to neutralize pathogens. Yet, viruses such as HIV can cause chronic infections by evolving as quickly as the adaptive immune system, forming an evolutionary arms race. Here we introduce a mathematical framework to study the coevolutionary dynamics between antibodies and antigens within a host. We focus on changes in the binding interactions between the antibody and antigen populations, which result from the underlying stochastic evolution of genotype frequencies driven by mutation, selection, and drift. We identify the critical viral and immune parameters that determine the distribution of antibody-antigen binding affinities. We also identify definitive signatures of coevolution that measure the reciprocal response between antibodies and viruses, and we introduce experimentally measurable quantities that quantify the extent of adaptation during continual coevolution of the two opposing populations. Using this analytical framework, we infer rates of viral and immune adaptation based on time-shifted neutralization assays in two HIV-infected patients. Finally, we analyze competition between clonal lineages of antibodies and characterize the fate of a given lineage in terms of the state of the antibody and viral populations. In particular, we derive the conditions that favor the emergence of broadly neutralizing antibodies, which may have relevance to vaccine design against HIV.
Rheumatoid Arthritis Disease Activity Is Determinant for ABCB1 and ABCG2 Drug-Efflux Transporters Function.Friday, July 22, 2016
Atisha-Fregoso Y, Lima G, Pascual-Ramos V, Baños-Peláez M, Fragoso-Loyo H, Jakez-Ocampo J, Contreras-Yáñez I, Llorente L,
PloS one. 2016
Patients with active RA have an increased function of ABCB1 and ABCG2, and disease activity is the main determinant of this phenomena.
Transcriptomes of whole blood and PBMC in chickens.Thursday, July 21, 2016
Désert C, Merlot E, Zerjal T, Bed'hom B, Härtle S, Le Cam A, Roux PF, Baeza E, Gondret F, Duclos MJ, Lagarrigue S,
Comparative biochemistry and physiology. Part D, Genomics & proteomics. 27-Jun-2016
Global transcriptome analysis of chicken whole blood to discover biomarkers of different phenotypes or physiological disorders has never been investigated so far. Whole blood provides significant advantages, allowing large scale and non-invasive sampling. However, generation of gene expression data from the blood of non-mammalian species remains a challenge, notably due to the nucleated red blood cells, hindering the use of well-established protocols. The aim of this study was to analyze the relevance of using whole blood cells (WB) to find biomarkers, instead of Peripheral Blood Mononuclear Cells (PBMC), usually chosen for immune challenges. RNA sources from WB and PBMC was characterized by microarray analysis. Our results show that the quality and quantity of RNA obtained from WB was suitable for further analyses, although the quality was lower than that from PBMC. The transcriptome profiling comparison revealed that the majority of genes were expressed in both WB and PBMC. Hemoglobin subunits were the major transcripts in WB, whereas the most enriched biological process was related to protein catabolic process. Most of the over-represented transcripts in PBMC were implicated in functions specific to thrombocytes, like coagulation and platelet activation, probably due to the large proportion of this nucleated cell type in chicken PBMC. Functions related to B and T cells and to other immune functions were also enriched in the PBMC subset. We conclude that WB is more suitable for large scale immunity oriented studies and other biological processes that have been poorly investigated so far.
Generation and Characterization of a Bivalent HIV-1 Subtype C gp120 Protein Boost for Proof-of-Concept HIV Vaccine Efficacy Trials in Southern Africa.Friday, July 22, 2016
Zambonelli C, Dey AK, Hilt S, Stephenson S, Go EP, Clark DF, Wininger M, Labranche C, Montefiori D, Liao HX, Swanstrom RI, Desaire H, Haynes BF, Carfi A, Barnett SW,
PloS one. 2016
The viral envelope glycoprotein (Env) is the major target for antibody (Ab)-mediated vaccine development against the Human Immunodeficiency Virus type 1 (HIV-1). Although several recombinant Env antigens have been evaluated in clinical trials, only the surface glycoprotein, gp120, (from HIV-1 subtype B, MN, and subtype CRF_01AE, A244) used in the ALVAC prime-AIDSVAX gp120 boost RV144 Phase III HIV vaccine trial was shown to contribute to protective efficacy, although modest and short-lived. Hence, for clinical trials in southern Africa, a bivalent protein boost of HIV-1 subtype C gp120 antigens composed of two complementary gp120s, from the TV1.C (chronic) and 1086.C (transmitted founder) HIV-1 strains, was selected. Stable Chinese Hamster Cell (CHO) cell lines expressing these gp120s were generated, scalable purification methods were developed, and a detailed analytical analysis of the purified proteins was conducted that showed differences and complementarity in the antigenicity, glycan occupancy, and glycan content of the two gp120 molecules. Moreover, mass spectrometry revealed some disulfide heterogeneity in the expressed proteins, particularly in V1V2-C1 region and most prominently in the TV1 gp120 dimers. These dimers not only lacked binding to certain key CD4 binding site (CD4bs) and V1V2 epitope-directed ligands but also elicited reduced Ab responses directed to those epitopes, in contrast to monomeric gp120, following immunization of rabbits. Both monomeric and dimeric gp120s elicited similarly high titer Tier 1 neutralizing Abs as measured in standard virus neutralization assays. These results provide support for clinical evaluations of bivalent preparations of purified monomeric TV1.C and 1086.C gp120 proteins.
Systematic reanalysis of clinical exome data yields additional diagnoses: implications for providers.Thursday, July 21, 2016
Wenger AM, Guturu H, Bernstein JA, Bejerano G,
Genetics in medicine : official journal of the American College of Medical Genetics. 21-Jul-2016
Approximately 250 gene-disease and 9,200 variant-disease associations are reported annually. This increase in information necessitates regular reevaluation of nondiagnostic exomes. To be practical, systematic reanalysis requires further automation and more up-to-date variant databases. To maximize the diagnostic yield of exome sequencing, providers should periodically request reanalysis of nondiagnostic exomes. Accordingly, policies regarding reanalysis should be weighed in combination with factors such as cost and turnaround time when selecting a clinical exome laboratory.Genet Med advance online publication 21 July 2016Genetics in Medicine (2016); doi:10.1038/gim.2016.88.
Insights into the biodegradation of weathered hydrocarbons in contaminated soils by bioaugmentation and nutrient stimulation.Thursday, July 21, 2016
Jiang Y, Brassington KJ, Prpich G, Paton GI, Semple KT, Pollard SJ, Coulon F,
Chemosphere. 18-Jul-2016
The potential for biotransformation of weathered hydrocarbon residues in soils collected from two commercial oil refinery sites (Soil A and B) was studied in microcosm experiments. Soil A has previously been subjected to on-site bioremediation and it was believed that no further degradation was possible while soil B has not been subjected to any treatment. A number of amendment strategies including bioaugmentation with hydrocarbon degrader, biostimulation with nutrients and soil grinding, were applied to the microcosms as putative biodegradation improvement strategies. The hydrocarbon concentrations in each amendment group were monitored throughout 112 days incubation. Microcosms treated with biostimulation (BS) and biostimulation/bioaugmentation (BS + BA) showed the most significant reductions in the aliphatic and aromatic hydrocarbon fractions. However, soil grinding was shown to reduce the effectiveness of a nutrient treatment on the extent of biotransformation by up to 25% and 20% for the aliphatic and aromatic hydrocarbon fractions, respectively. This is likely due to the disruption to the indigenous microbial community in the soil caused by grinding. Further, ecotoxicological responses (mustard seed germination and Microtox assays) showed that a reduction of total petroleum hydrocarbon (TPH) concentration in soil was not directly correlable to reduction in toxicity; thus monitoring TPH alone is not sufficient for assessing the environmental risk of a contaminated site after remediation.
Diagnostic significance of suppressor of cytokine signalling 3 (SOCS3) methylation and its correlation with IDH1 mutation in Chinese glioma patients.Thursday, July 21, 2016
Jiao W, Xun X, Liu J, Yang J, Wang Q, Wang L, Chen C, Wang H, Dai P,
Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals. 21-Jul-2016
SOCS3 methylation is a potential biomarker for grading and prognosis in human glioma.
An Algorithm for Systemic Inflammatory Response Syndrome Criteria-Based Prediction of Sepsis in a Polytrauma Cohort.Thursday, July 21, 2016
Lindner HA, Balaban Ü, Sturm T, Wei C, Thiel M, Schneider-Lindner V,
Critical care medicine. 15-Jul-2016
Dynamic systemic inflammatory response syndrome descriptors improved specificity of sepsis prediction and particularly diagnosis, rivaling established biomarkers, in a polytrauma cohort. They may enhance electronic sepsis surveillance once evaluated in other patient populations.
Platelet Count Trends and Prevalence of Heparin-Induced Thrombocytopenia in a Cohort of Extracorporeal Membrane Oxygenator Patients.Thursday, July 21, 2016
Sokolovic M, Pratt AK, Vukicevic V, Sarumi M, Johnson LS, Shah NS,
Critical care medicine. 15-Jul-2016
Prevalence of heparin-induced thrombocytopenia and heparin-induced thrombocytopenia-related thrombosis among extracorporeal membrane oxygenator patients at our institution is relatively high. Using platelet count trends to guide decision to test for heparin-induced thrombocytopenia is not an optimal strategy in extracorporeal membrane oxygenator patients. Without a validated pretest probability clinical score, serosurveillance in a defined high-risk group of extracorporeal membrane oxygenator patients may be needed.
Bioactive Dibenzo-α-pyrone Derivatives from the Endophytic Fungus Rhizopycnis vagum Nitaf22.Thursday, July 21, 2016
Lai D, Wang A, Cao Y, Zhou K, Mao Z, Dong X, Tian J, Xu D, Dai J, Peng Y, Zhou L, Liu Y,
Journal of natural products. 21-Jul-2016
Six new dibenzo-α-pyrones, rhizopycnolides A (1) and B (2) and rhizopycnins A-D (3-6), together with eight known congeners (7-14), were isolated from the endophytic fungus Rhizopycnis vagum Nitaf22 obtained from Nicotiana tabacum. The structures of the new compounds were unambiguously elucidated using NMR, HRESIMS, TDDFT ECD calculation, and X-ray crystallography data. Rhizopycnolides A (1) and B (2) feature an uncommon γ-butyrolactone-fused dibenzo-α-pyrone tetracyclic skeleton (6/6/6/5), while rhizopycnin B (4) was the first amino group containing dibenzo-α-pyrone. Rhizopycnolides A (1) and B (2) are proposed to be biosynthesized from polyketide and tricarboxylic acid cycle pathways. The isolated compounds were tested for their antibacterial, antifungal, and cytotoxic activities. Among them, rhizopycnolide A (1), rhizopycnins C (5) and D (6), TMC-264 (8), penicilliumolide D (11), and alternariol (12) were active against the tested pathogenic bacteria Agrobacterium tumefaciens, Bacillus subtilis, Pseudomonas lachrymans, Ralstonia solanacearum, Staphylococcus hemolyticus, and Xanthomonas vesicatoria with MIC values in the range 25-100 μg/mL. Rhizopycnin D (6) and TMC-264 (8) strongly inhibited the spore germination of Magnaporthe oryzae with IC50 values of 9.9 and 12.0 μg/mL, respectively. TMC-264 (8) showed potent cytotoxicity against five human cancer cell lines (HCT-116, HepG2, BGC-823, NCI-H1650, and A2780) with IC50 values of 3.2-7.8 μM.
Survival of Helicobacter pylori in the wastewater treatment process and the receiving river in Michigan, USA.Friday, July 22, 2016
Bai X, Xi C, Wu J,
Journal of water and health. Aug-2016
Contaminated water may play a key role in the transmission of Helicobacter pylori, resulting in gastrointestinal diseases in humans. The wastewater treatment process is an important barrier to control the transmission of H. pylori. However, the presence and viability of H. pylori in the treatment process is not well known. In this paper, the real colony morphology of H. pylori was confirmed by two types of culture media. The survival of H. pylori through the tertiary wastewater treatment process, especially UV disinfection, and in the receiving Huron River in Ann Arbor, Michigan, was investigated by plates cultivation, regular polymerase chain reaction (PCR) assays and quantitative real-time PCR from DNA. The results demonstrated that H. pylori was not only present, but also viable in all processed wastewater samples in the Ann Arbor wastewater treatment plant (WWTP). H. pylori can be found in a higher concentration in the receiving Huron River. There are many kinds of antibiotic- and UV-resistant bacteria, including H. pylori, in the final effluent of Ann Arbor WWTP.
The effect of chlorine and combined chlorine/UV treatment on coliphages in drinking water disinfection.Friday, July 22, 2016
Zyara AM, Torvinen E, Veijalainen AM, Heinonen-Tanski H,
Journal of water and health. Aug-2016
Chlorine disinfection is a globally used method to ensure the safety of drinking water. However, it has not always been successful against viruses and, therefore, it is important to find new methods to disinfect water. Seventeen different coliphages were isolated from the treated municipal wastewater. These coliphages and MS2 were treated with different dosages of chlorine in drinking water, and a combined chlorine/ultraviolet irradiation treatment for the chlorine-resistant coliphages. Chlorine disinfection with 0.3-0.5 mg/L total chlorine (free Cl-dosage 0.12-0.21 mg/L) for 10 min achieved 2.5-5.7 Log10-reductions for 11 sensitive coliphages. The six most resistant coliphages showed no reduction with these chlorine concentrations. MS2 was intermediate in chlorine resistance, and thus it is not a good indicator for viruses in chlorine disinfection. In the combined treatment total chlorine of 0.05-0.25 mg/L (free Cl-dosage 0.02-0.08 mg/L) and ultraviolet irradiation (14-22 mWs/cm(2)) were more effective than chlorine alone, and 3-5 Log10-reductions were achieved for the chlorine-resistant strains. The chlorination efficiency could be increased by higher dosages and longer contact times, but this could increase the formation of disinfection by-products. Therefore, the combination treatment is a recommended disinfection method.
Comparison of the Performances of Five Primer Sets for the Detection and Quantification of Plasmodium in Anopheline Vectors by Real-Time PCR.Friday, July 22, 2016
Chaumeau V, Andolina C, Fustec B, Tuikue Ndam N, Brengues C, Herder S, Cerqueira D, Chareonviriyaphap T, Nosten F, Corbel V,
PloS one. 2016
Quantitative real-time polymerase chain reaction (qrtPCR) has made a significant improvement for the detection of Plasmodium in anopheline vectors. A wide variety of primers has been used in different assays, mostly adapted from molecular diagnosis of malaria in human. However, such an adaptation can impact the sensitivity of the PCR. Therefore we compared the sensitivity of five primer sets with different molecular targets on blood stages, sporozoites and oocysts standards of Plasmodium falciparum (Pf) and P. vivax (Pv). Dilution series of standard DNA were used to discriminate between methods at low concentrations of parasite and to generate standard curves suitable for the absolute quantification of Plasmodium sporozoites. Our results showed that the best primers to detect blood stages were not necessarily the best ones to detect sporozoites. Absolute detection threshold of our qrtPCR assay varied between 3.6 and 360 Pv sporozoites and between 6 and 600 Pf sporozoites per mosquito according to the primer set used in the reaction mix. In this paper, we discuss the general performance of each primer set and highlight the need to use efficient detection methods for transmission studies.
Circulating miR-223 in Oral Cancer: Its Potential as a Novel Diagnostic Biomarker and Therapeutic Target.Friday, July 22, 2016
Tachibana H, Sho R, Takeda Y, Zhang X, Yoshida Y, Narimatsu H, Otani K, Ishikawa S, Fukao A, Asao H, Iino M,
PloS one. 2016
Circulating microRNAs (miRNAs) have been detected in various types of cancer and have been proposed as novel biomarkers for diagnosis and treatment. Until recently, however, no studies had comprehensively examined circulating miRNAs in oral cancer. The current study used an ultra-sensitive genome-wide miRNA array to investigate changes in circulating miRNAs in plasma from five patients with oral cancer and ten healthy individuals. Results indicated that there were only a few circulating miRNAs, including miR-223, miR-26a, miR-126, and miR-21, that were up-regulated in patients with oral cancer. A subsequent validation test indicated that circulating miR-223 levels were significantly higher (~2-fold, P< 0.05) in patients with oral cancer (n = 31) than in those without cancer (n = 31). Moreover, miR-223 was found to be up-regulated in tumor-adjacent normal tissue compared to tumor tissue from patients with oral cancer. A gain-of-function assay was performed to explore the potential roles of circulating miR-223 in the development of oral cancer. Results revealed that miR-223 functions as a tumor suppressor by inhibiting cell proliferation and inducing apoptosis. In conclusion, this study suggested that circulating miR-223 may serve as a potential biomarker for diagnosis and that it may represent a novel therapeutic target for treatment of oral cancer.
Direct Quantification of DNA Base Composition by Surface-Enhanced Raman Scattering Spectroscopy.Tuesday, July 26, 2016
Morla-Folch J, Alvarez-Puebla RA, Guerrini L,
The journal of physical chemistry letters. 26-Jul-2016
Design of ultrasensitive DNA sensors based on the unique physical properties of plasmonic nanostructures has become one of the most exciting areas in nanomedicine. However, despite the vast number of proposed applications, the determination of the base composition in nucleic acids, a fundamental parameter in genomic analyses and taxonomic classification, is still restricted to time-consuming and poorly sensitive conventional methods. Herein, we demonstrate the possibility of determining the base composition in single- and double-stranded DNA by using a simple, low-cost, high-throughput, and label-free surface-enhanced Raman scattering (SERS) method in combination with cationic nanoparticles.
Stereotactic Body Radiotherapy for Oligometastasis: Opportunities for Biology to Guide Clinical Management.Friday, July 22, 2016
Correa RJ, Salama JK, Milano MT, Palma DA,
Cancer journal (Sudbury, Mass.). 26-7-2016
Oligometastasis refers to a state of limited metastatic disease burden, in which surgical or ablative treatment to all known visible metastases holds promise to extend survival or even effect cure. Stereotactic body radiotherapy is a form of radiation treatment capable of delivering a high biologically effective dose of radiation in a highly conformal manner, with a favorable toxicity profile. Enthusiasm for oligometastasis ablation, however, should be counterbalanced against the limited supporting evidence. It remains unknown to what extent (if any) ablation influences survival or quality of life. Rising clinical equipoise necessitates the completion of randomized controlled trials to assess this, several of which are underway. However, a lack of clear identification criteria or biomarkers to define the oligometastatic state hampers optimal patient selection.This narrative review explores the evolutionary origins of oligometastasis, the steps of the metastatic process at which oligometastases may arise, and the biomolecular mediators of this state. It discusses clinical outcomes with treatment of oligometastases, ongoing trials, and areas of basic and translational research that may lead to novel biomarkers. These efforts should provide a clearer, biomolecular definition of oligometastatic disease and aid in the accurate selection of patients for ablative therapies.
Rinsing with Saline Promotes Human Gingival Fibroblast Wound Healing In Vitro.Friday, July 22, 2016
Huynh NC, Everts V, Leethanakul C, Pavasant P, Ampornaramveth RS,
PloS one. 2016
Rinsing the mouth with sodium chloride (NaCl) solution is believed to promote healthy gum and improve oral ulcer healing. Scientific evidence to support this assumption is, however, lacking. This study aims to investigate the effect and clarify underlying mechanisms of short-term rinsing with NaCl on human gingival fibroblast (hGFs) wound healing. Isolated primary hGFs and human normal oral keratinocytes (hNOKs) were rinsed with 0-7.2% NaCl for 2 min, 3 times a day. Scratch-tests, trans-well migration assays and MTT activity were performed. mRNA expression was assessed of type-I collagen, fibronectin and FAK. Changes in FAK and F-actin were detected by immunofluorescence. KCl, NaH2PO4, KH2PO4 were used to clarify the molecules involved. Rinsing with 0.9-1.8% NaCl significantly promoted hGFs cell migration but not proliferation. However, it had no effect on hNOKs. Rinsing with 1.8% NaCl significantly up-regulated the expression of type-I collagen and fibronectin. FAK and F-actin, molecules responsible for cytoskeleton re-organization and cell migration, were also up-regulated. Cl- seemed to be essential since rinsing with KCl resulted in a similar effect as noted with NaCl. In conclusion, short-term rinsing with NaCl promoted hGFs migration, and increased the expression of extracellular matrix as well as cytoskeletal proteins. These data strongly support the long held belief in the benefits of using NaCl mouth-rinse.
Improving the Efficiency and Ease of Healthcare Analysis Through Use of Data Visualization Dashboards.Friday, July 22, 2016
Stadler JG, Donlon K, Siewert JD, Franken T, Lewis NE,
Big data. Jun-2016
The digitization of a patient's health record has profoundly impacted medicine and healthcare. The compilation and accessibility of medical history has provided clinicians an unprecedented, holistic account of a patient's conditions, procedures, medications, family history, and social situation. In addition to the bedside benefits, this level of information has opened the door for population-level monitoring and research, the results of which can be used to guide initiatives that are aimed at improving quality of care. Cerner Corporation partners with health systems to help guide population management and quality improvement projects. With such an enormous and diverse client base-varying in geography, size, organizational structure, and analytic needs-discerning meaning in the data and how they fit with that particular hospital's goals is a slow, difficult task that requires clinical, statistical, and technical literacy. This article describes the development of dashboards for efficient data visualization at the healthcare facility level. Focusing on two areas with broad clinical importance, sepsis patient outcomes and 30-day hospital readmissions, dashboards were developed with the goal of aggregating data and providing meaningful summary statistics, highlighting critical performance metrics, and providing easily digestible visuals that can be understood by a wide range of personnel with varying levels of skill and areas of expertise. These internal-use dashboards have allowed associates in multiple roles to perform a quick and thorough assessment on a hospital of interest by providing the data to answer necessary questions and to identify important trends or opportunities. This automation of a previously manual process has greatly increased efficiency, saving hours of work time per hospital analyzed. Additionally, the dashboards have standardized the analysis process, ensuring use of the same metrics and processes so that overall themes can be compared across hospitals and health systems.
Who Watches the Watchmen: Roles of RNA Modifications in the RNA Interference Pathway.Friday, July 22, 2016
Shelton SB, Reinsborough C, Xhemalce B,
PLoS genetics. Jul-2016
RNA levels are widely thought to be predictive of RNA function. However, the existence of more than a hundred chemically distinct modifications of RNA alone is a major indication that these moieties may impart distinct functions to subgroups of RNA molecules that share a primary sequence but display distinct RNA "epigenetic" marks. RNAs can be modified on many sites, including 5' and 3' ends, the sugar phosphate backbone, or internal bases, which collectively provide many opportunities for posttranscriptional regulation through a variety of mechanisms. Here, we will focus on how modifications on messenger and microRNAs may affect the process of RNA interference in mammalian cells. We believe that taking RNA modifications into account will not only advance our understanding of this crucial pathway in disease and cancer but will also open the path to exploiting the enzymes that "write" and "erase" them as targets for therapeutic drug development.
Size- and coating-dependent cytotoxicity and genotoxicity of silver nanoparticles evaluated using in vitro standard assays.Thursday, July 21, 2016
Guo X, Li Y, Yan J, Ingle T, Jones MY, Mei N, Boudreau MD, Cunningham CK, Abbas M, Paredes A, Zhou T, Moore MM, Howard PC, Chen T,
Nanotoxicology. 21-Jul-2016
The physicochemical characteristics of silver nanoparticles (AgNPs) may greatly alter their toxicological potential. To explore the effects of size and coating on the cytotoxicity and genotoxicity of AgNPs, six different types of AgNPs, having 3 different sizes and 2 different coatings, were investigated using the Ames test, mouse lymphoma assay (MLA), and in vitro micronucleus assay. The genotoxicities of silver acetate and silver nitrate were evaluated to compare the genotoxicity of nanosilver to that of ionic silver. The Ames test produced inconclusive results for all types of the silver materials due to the high toxicity of silver to the test bacteria and the lack of entry of the nanoparticles into the cells. Treatment of L5718Y cells with AgNPs and ionic silver resulted in concentration-dependent cytotoxicity, mutagenicity in the Tk gene and the induction of micronucleus from exposure to nearly every type of the silver materials. Treatment of TK6 cells with these silver materials also resulted in concentration dependent cytotoxicity and significantly increased micronucleus frequency. In both the MLA and micronucleus assays the smaller AgNPs, the higher the cytotoxicity and genotoxicity. The coatings had less effect on the relative genotoxicity of AgNPs than the particle size. Loss of heterozygosity analysis of the induced Tk mutants indicated that the types of mutations induced by AgNPs were different from those of ionic silver. These results suggest that AgNPs induce cytotoxicity and genotoxicity in a size- and coating-dependent manner. Furthermore, while the MLA and in vitro micronucleus assay (in both types of cells) are useful to quantitatively measure the genotoxic potencies of AgNPs, the Ames test cannot.
Assessment of Risk and Sero-Prevalence of Helicobacter pylori Colonization among Remote Orang Asli Tribes in Peninsula Malaysia.Friday, July 22, 2016
Thevakumar K, Chandren JR, Perez-Perez GI, Chua EG, Teh LK, Salleh MZ, Tan JA, Leow AH, Goh KL, Tay AC, Marshall BJ, Vadivelu J, Loke MF, Wong LP,
PloS one. 2016
The epidemiology of Helicobacter pylori (H. pylori) infection is related to human poverty with marked differences between developing and developed countries. Socioeconomic factors and living standards are the main determinants of the age-dependent acquisition rate of H. pylori, and consequently its prevalence. The aim of this study was to assess the risk and sero-prevalence of H. pylori colonization among Orang Asli in Peninsula Malaysia. This cross-sectional study was conducted on Orang Asli subjects in seven isolated settlements spanning across all three major tribes (Negrito, Proto Malay and Senoi) in Malaysia. Socio-demographic characteristics of the subjects were obtained through interview. Subjects were tested for H. pylori colonization based on CagA and whole cell (WC) antigen serological assays. A total of 275 subjects participated in this study. Among these subjects, 115 (44.7%) were H. pylori sero-positive with highest sero-prevalence among Negrito (65.7%). Among subjects who were H. pylori sero-positive, CagA sero positivity was also significantly higher among Negrito. The highest proportion of respondents reported to be H. pylori sero-positive was from age group 30 years old and below (57.9%), males (56.2%), Negrito (48.6%) and live in bamboo house (92.3%). The highest proportion of respondents reported to be CagA sero-positive was from age group 30 years old and below (41.4%), males (35.6%) and Negrito (48.6%). The results of this study demonstrate that H. pylori colonization can be related to age, gender, tribes and house materials and CagA sero-positive stain closely associated with age, gender and tribes.
The Immune Landscapes of Polypoid and Nonpolypoid Precancerous Colorectal Lesions.Friday, July 22, 2016
Maglietta A, Maglietta R, Staiano T, Bertoni R, Ancona N, Marra G, Resta L,
PloS one. 2016
Little is known about the immunoediting process in precancerous lesions. We explored this aspect of benign colorectal adenomas with a descriptive analysis of the immune pathways and immune cells whose regulation is linked to the morphology and size of these lesions. Two series of polypoid and nonpolypoid colorectal adenomas were used in this study: 1) 84 samples (42 lesions, each with matched samples of normal mucosa) whose gene expression data were used to quantify the tumor morphology- and size-related dysregulation of immune pathways collected in the Molecular Signature Database, using Gene Set Enrichment Analysis; 2) 40 other lesions examined with immunohistochemistry to quantify the presence of immune cells in the stromal compartment. In the analysis of transcriptomic data, 429 immune pathways displayed significant differential regulation in neoplasms of different morphology and size. Most pathways were significantly upregulated or downregulated in polypoid lesions versus nonpolypoid lesions (regardless of size). Differential pathway regulation associated with lesion size was observed only in polypoid neoplasms. These findings were mirrored by tissue immunostaining with CD4, CD8, FOXP3, MHC-I, CD68, and CD163 antibodies: stromal immune cell counts (mainly T lymphocytes and macrophages) were significantly higher in polypoid lesions. Certain markers displayed significant size-related differences regardless of lesion morphology. Multivariate analysis of variance showed that the marker panel clearly discriminated between precancerous lesions of different morphologies and sizes. Statistical analysis of immunostained cell counts fully support the results of the transcriptomic data analysis: the density of infiltration of most immune cells in the stroma of polypoid precancerous lesions was significantly higher than that observed in nonpolypoid lesions. Large neoplasms also have more immune cells in their stroma than small lesions. Immunoediting in precancerous colorectal tumors may vary with lesion morphology and stage of development, and this variability could influence a given lesion's trajectory to cancer.
Samura B,
Georgian medical news. Jun-2016
to investigate an interrelationship between pre-treatment galcetin-3 (Gal-3) level and one-year survival rate, cardiovascular events in subjects with multiple myeloma. Eighty nine subjects with full or partial remission of multiple myeloma were enrolled in the study. Patients were divided into 2 groups based on whether or not statins were included in their treatment: a statin group (n=43) and a no statin group (n=46). Among the 43 patients in the statin group, 31 patients received 20mg/day atorvastatin and 12 patients received 40-mg/day atorvastatin. None of the patients had received any lipid-modulating medications, including statins or fibrates, before enrollment. Observation period was up to 1 year. Blood samples for biomarkers measurements were collected. ELISA method for measurements of circulating level of galectin-3, interleukin-6 and NT-pro-brain natriuretic peptide were used. Lipid lowering effect in statin user was associates with declined serum Gal-3 level, whereas in not statin users similar response was not appeared. No any changes in hemodynamics and other biomarkers between both cohorts were found. Univariate logistic regression had exhibited that galtctin-3 (odds ratio [OR] = 1.17; 95% CI = 1.07-1.29; P = 0.002), NT-pro-brain natriuretic peptide (OR=1.04; 95% CI=1.02-1.10; P<0.05) and statin therapy (OR=1.07; 95% CI = 1.02-1.11; P = 0.001) predicted one-year cumulative CV events. After adjustment on statin therapy, galectin-3 remained independent predictor one-year cumulative cardiovascular events (OR=1.08; 95% CI=1.06-1.11; P=0.001). When initial serum galectin-3 level has incorporated into prediction model, statin therapy was found as predictor for improving survival in multiple myeloma patients with elevated serum galectin-3 level (>14 ng/ml). Elevated pre-treatment galectin-3 level was found a powerful predictor of positive effect of statins on survival in patients with regression of multiple myeloma.
Tuleubaev B, Saginova D, Abiyev T, Davletbaev M, Koshanova A,
Georgian medical news. Jun-2016
Despite the variety of treatments available, including surgical procedures and antimicrobial therapy, bone infections is still a medical problem, because they are difficult to treat. Optimal treatment should stabilize the bone, promote the biological recovery of bone defects and destroy bacterial infection. Systemic antibiotics are part of the standard therapy after surgical treatment of infected bone, but their effectiveness is limited due to malnutrition and low absorption at the site of infection. Moreover, long-term treatment and higher doses are associated with serious side effects. In contrast, the antibiotic impregnated bone cements or fillers can act as a local anti-infective drug delivery system, which not only fills the dead space after debridement, but also provide high concentrations of antibiotics in a potential site of infection, no increase levels of antibiotics in serum. The review analyzed the use of antibiotic-impregnated cement as local delivery of antibiotics systems. Gentamycin impregnated polymethylmethacrylate (PMMA) beads, for the topical treatment of orthopedic infections clinically used for over 30 years. Application of antibiotic delivery systems using cement in the infected region is common method of treatment that continues to improve. On the downside of PMMA is that the material does not biodegradable requires subsequent invasive procedures necessary to remove the implant.
A Call to Action for Hazardous Drug Safety: Where We Have Been and Where We Are Now.Friday, July 22, 2016
Eisenberg S,
Clinical journal of oncology nursing. 1-Aug-2016
Although improvements have been made in the use of personal protective equipment, studies indicate that nurses continue to be unnecessarily at risk. Inability to fully understand the dangers, a lack of organizational safety culture, and the general inability to enforce guidelines continue to be challenging. Fortunately, a number of upcoming changes will help to build momentum for increasing nursing safety.
Expression of miR-210 in relation to other measures of hypoxia and prediction of benefit from hypoxia modification in patients with bladder cancer.Thursday, July 21, 2016
Irlam-Jones JJ, Eustace A, Denley H, Choudhury A, Harris AL, Hoskin PJ, West CM,
British journal of cancer. 21-Jul-2016
High miR-210 may reflect hypoxia in bladder cancer. However, its ability to predict benefit from hypoxia modification does not improve upon other hypoxia markers. Investigation as part of a miRNA hypoxia signature may reveal the full potential of miR-210.British Journal of Cancer advance online publication, 21 July 2016; doi:10.1038/bjc.2016.218
Genetic progression of Barrett's oesophagus to oesophageal adenocarcinoma.Thursday, July 21, 2016
Gregson EM, Bornschein J, Fitzgerald RC,
British journal of cancer. 21-Jul-2016
Barrett's oesophagus (BE) is the premalignant condition associated with the development of oesophageal adenocarcinoma (OAC). Diagnostically, p53 immunohistochemistry remains the only biomarker recommended clinically to aid histopathological diagnosis. The emerging mutational profile of BE is one of highly heterogeneous lesions at the genomic level with many mutations already occurring in non-dysplastic tissue. As well as point mutations, larger scale copy-number changes appear to have a key role in the progression to OAC and clinically applicable assays for the reliable detection of aneuploidy will be important to incorporate into future clinical management of patients. For some patients, the transition to malignancy may occur rapidly through a genome-doubling event or chromosomal catastrophe, termed chromothripsis, and detecting these patients may prove especially difficult. Given the heterogeneous nature of this disease, sampling methods to overcome inherent bias from endoscopic biopsies coupled with the development of more objective biomarkers than the current reliance on histopathology will be required for risk stratification. The aim of this approach will be to spare low-risk patients unnecessary procedures, as well as to provide endoscopic therapy to the patients at highest risk, thereby avoiding the burden of incurable metastatic disease.British Journal of Cancer advance online publication, 21 July 2016; doi:10.1038/bjc.2016.219
Microdialysis Sampling from Wound Fluids Enables Quantitative Assessment of Cytokines, Proteins, and Metabolites Reveals Bone Defect-Specific Molecular Profiles.Friday, July 22, 2016
Förster Y, Schmidt JR, Wissenbach DK, Pfeiffer SE, Baumann S, Hofbauer LC, von Bergen M, Kalkhof S, Rammelt S,
PloS one. 2016
Bone healing involves a variety of different cell types and biological processes. Although certain key molecules have been identified, the molecular interactions of the healing progress are not completely understood. Moreover, a clinical routine for predicting the quality of bone healing after a fracture in an early phase is missing. This is mainly due to a lack of techniques to comprehensively screen for cytokines, growth factors and metabolites at their local site of action. Since all soluble molecules of interest are present in the fracture hematoma, its in-depth assessment could reveal potential markers for the monitoring of bone healing. Here, we describe an approach for sampling and quantification of cytokines and metabolites by using microdialysis, combined with solid phase extractions of proteins from wound fluids. By using a control group with an isolated soft tissue wound, we could reveal several bone defect-specific molecular features. In bone defect dialysates the neutrophil chemoattractants CXCL1, CXCL2 and CXCL3 were quantified with either a higher or earlier response compared to dialysate from soft tissue wound. Moreover, by analyzing downstream adaptions of the cells on protein level and focusing on early immune response, several proteins involved in the immune cell migration and activity could be identified to be specific for the bone defect group, e.g. immune modulators, proteases and their corresponding inhibitors. Additionally, the metabolite screening revealed different profiles between the bone defect group and the control group. In summary, we identified potential biomarkers to indicate imbalanced healing progress on all levels of analysis.
Ajwa Date (Phoenix dactylifera L.) Extract Inhibits Human Breast Adenocarcinoma (MCF7) Cells In Vitro by Inducing Apoptosis and Cell Cycle Arrest.Friday, July 22, 2016
Khan F, Ahmed F, Pushparaj PN, Abuzenadah A, Kumosani T, Barbour E, AlQahtani M, Gauthaman K,
PloS one. 2016
MEAD inhibited MCF7 cells in vitro by the inducing cell cycle arrest and apoptosis. Our results indicate the anticancer effects of Ajwa dates, which therefore may be used as an adjunct therapy with conventional chemotherapeutics to achieve a synergistic effect against breast cancer.
Molecular Mechanisms for Drug Hypersensitivity Induced by the Malaria Parasite's Chloroquine Resistance Transporter.Friday, July 22, 2016
Richards SN, Nash MN, Baker ES, Webster MW, Lehane AM, Shafik SH, Martin RE,
PLoS pathogens. Jul-2016
Mutations in the Plasmodium falciparum 'chloroquine resistance transporter' (PfCRT) confer resistance to chloroquine (CQ) and related antimalarials by enabling the protein to transport these drugs away from their targets within the parasite's digestive vacuole (DV). However, CQ resistance-conferring isoforms of PfCRT (PfCRTCQR) also render the parasite hypersensitive to a subset of structurally-diverse pharmacons. Moreover, mutations in PfCRTCQR that suppress the parasite's hypersensitivity to these molecules simultaneously reinstate its sensitivity to CQ and related drugs. We sought to understand these phenomena by characterizing the functions of PfCRTCQR isoforms that cause the parasite to become hypersensitive to the antimalarial quinine or the antiviral amantadine. We achieved this by measuring the abilities of these proteins to transport CQ, quinine, and amantadine when expressed in Xenopus oocytes and complemented this work with assays that detect the drug transport activity of PfCRT in its native environment within the parasite. Here we describe two mechanistic explanations for PfCRT-induced drug hypersensitivity. First, we show that quinine, which normally accumulates inside the DV and therewithin exerts its antimalarial effect, binds extremely tightly to the substrate-binding site of certain isoforms of PfCRTCQR. By doing so it likely blocks the normal physiological function of the protein, which is essential for the parasite's survival, and the drug thereby gains an additional killing effect. In the second scenario, we show that although amantadine also sequesters within the DV, the parasite's hypersensitivity to this drug arises from the PfCRTCQR-mediated transport of amantadine from the DV into the cytosol, where it can better access its antimalarial target. In both cases, the mutations that suppress hypersensitivity also abrogate the ability of PfCRTCQR to transport CQ, thus explaining why rescue from hypersensitivity restores the parasite's sensitivity to this antimalarial. These insights provide a foundation for understanding clinically-relevant observations of inverse drug susceptibilities in the malaria parasite.
Short-term effects of cannabidiol after global hypoxia-ischemia in newborn piglets.Thursday, July 21, 2016
Garberg HT, Huun MU, Escobar J, Martinez-Orgado J, Løberg EM, Solberg R, Saugstad OD,
Pediatric research. 21-Jul-2016
In contrast to previous studies, we do not find significant protective effects of CBD after HI in piglets. Evaluation of CBD in higher doses might be warranted.
Lin JS, Piper MA, Perdue LA, Rutter C, Webber EM, O’Connor E, Smith N, Whitlock EP,
Where is it and how much? Mapping and quantifying elements in single cells.Thursday, July 21, 2016
Malucelli E, Fratini M, Notargiacomo A, Gianoncelli A, Merolle L, Sargenti A, Cappadone C, Farruggia G, Lagomarsino S, Iotti S,
The Analyst. 21-Jul-2016
The biological function of a chemical element in cells not only requires the determination of its intracellular quantity, but also the spatial distribution of its concentration. Different strategies can be employed to quantify and map the intracellular concentration of elements in single cells. The assessment of the intracellular elemental concentration, which is the relevant information, requires the measurement of cell volume. This challenging and demanding task requires combining different techniques allowing gathering of both morphological and compositional information on the same cell. Moreover, the need to analyse samples more similar to their natural state requires complex hardware equipment, and supplementary efforts in preparation protocols. Nevertheless, the response to the question: "where is it and how much?" is worth all these efforts. This review aims at providing an insight into the recent and most advanced techniques and strategies for quantifying and mapping chemical elements in single cells. We describe and discuss indirect detection techniques (label based) which make use of fluorescent dyes, and direct ones (label free), such as particle induced X-ray emission, proton backscattering spectrometry, scanning transmission ion spectrometry, nano-secondary ion mass spectrometry, X-ray fluorescence microscopy, complemented by X-ray imaging.
Screening for post 32-week preterm birth risk: how helpful is routine perinatal data collection?Thursday, July 21, 2016
Luo W, Huning EY, Tran T, Phung D, Venkatesh S,
Heliyon. Jun-2016
For multiparous women, the routine data contains information comparable to some purposely-collected data for predicting preterm risk. But for nulliparous women, the routine data contains insufficient data related to antenatal complications.
Influence of starter culture and a protease on the generation of ACE-inhibitory and antioxidant bioactive nitrogen compounds in Iberian dry-fermented sausage "salchichón".Thursday, July 21, 2016
Fernández M, Benito MJ, Martín A, Casquete R, Córdoba JJ, Córdoba MG,
Heliyon. Mar-2016
The effect of the addition of an autochthonous starter culture and the protease EPg222 on the generation of angiotensin-I-converting enzyme (ACE)-inhibitory and antioxidant compounds by the dry-fermented sausage "salchichón" was investigated. Sausages were prepared with purified EPg222 and Pediococcus acidilactici MS200 and Staphylococcus vitulus RS34 as the starter culture (P200S34), separately and together, ripened for 90 days, and compared to a control batch. Among the ripening time points (20, 35, 65, 90 days) studied, batches inoculated with EPg222 had higher nitrogen compound concentrations at 63 days of ripening. ACE-inhibitory and antioxidant activities were also highest in both batches with EPg222 at 63 days of ripening, and these activities were stable in most cases after in vitro simulated gastrointestinal digestion. These activities were correlated with the most relevant compounds detected by HLPC-ESI-MS. The principal components analysis (PCA) linked the P200S34 + EPg222 batch with the major compounds identified. The antioxidant activity was higher at 63 days of ripening, especially in highly proteolytic batches, such as P200S34 + EPg222. The ACE-inhibitory activity was not associated with any of the major compounds. The use of the enzyme EPg222 in association with the starter culture P200S34 in the preparation of dry-cured meat products could be of great importance due to their demonstrated ability to produce compounds with high biological activity, such as ACE-inhibitory and antioxidant activity.
Inhibitory effect of isoamericanol A from Jatropha curcas seeds on the growth of MCF-7 human breast cancer cell line by G2/M cell cycle arrest.Thursday, July 21, 2016
Katagi A, Sui L, Kamitori K, Suzuki T, Katayama T, Hossain A, Noguchi C, Dong Y, Yamaguchi F, Tokuda M,
Heliyon. Jan-2016
Although various parts of J. curcas (Jatropha curcas L., Euphorbiaceae) have long been used as traditional folk medicines for their antiviral, analgesic, and/or antidotal efficacies, we are the first to investigate the role of anti-carcinogenicity of isoamericanol A (IAA) from the seed extract. Our results showed that IAA is capable of inhibiting cell proliferation in a dose-dependent manner on the human cancer cell lines of MCF-7, MDA-MB231, HuH-7, and HeLa. Flow cytometry analysis showed IAA significantly induces cell cycle arrest at G2/M on MCF-7 cells. At both protein and mRNA levels examined by western blot and real-time PCR, the results revealed increased expression of BTG2 (B-cell translocation gene 2), p21 (p21(WAF1/CIPI) ), and GADD45A (growth arrest and DNA-damage-inducible, alpha) after IAA treatment, but inversed expression in CDK1 (cyclin-dependent kinase 1) and cyclins B1 and B2. All these effects contribute to G2/M cell cycle arrest. Furthermore, these results coincide with the changes in molecular expressions determined by DNA-microarray analysis. Our findings indicate that IAA has an inhibitory effect on cell proliferation of MCF-7 through cell cycle arrest, giving it great potential as a future therapeutic reagent for cancers.
Comprehensive characterization of evolutionary conserved breakpoints in four New World Monkey karyotypes compared to Chlorocebus aethiops and Homo sapiens.Thursday, July 21, 2016
Fan X, Supiwong W, Weise A, Mrasek K, Kosyakova N, Tanomtong A, Pinthong K, Trifonov VA, Cioffi Mde B, Grothmann P, Liehr T, Oliveira EH,
Heliyon. Nov-2015
Comparative cytogenetic analysis in New World Monkeys (NWMs) using human multicolor banding (MCB) probe sets were not previously done. Here we report on an MCB based FISH-banding study complemented with selected locus-specific and heterochromatin specific probes in four NWMs and one Old World Monkey (OWM) species, i.e. in Alouatta caraya (ACA), Callithrix jacchus (CJA), Cebus apella (CAP), Saimiri sciureus (SSC), and Chlorocebus aethiops (CAE), respectively. 107 individual evolutionary conserved breakpoints (ECBs) among those species were identified and compared with those of other species in previous reports. Especially for chromosomal regions being syntenic to human chromosomes 6, 8, 9, 10, 11, 12 and 16 previously cryptic rearrangements could be observed. 50.4% (54/107) NWM-ECBs were colocalized with those of OWMs, 62.6% (62/99) NWM-ECBs were related with those of Hylobates lar (HLA) and 66.3% (71/107) NWM-ECBs corresponded with those known from other mammalians. Furthermore, human fragile sites were aligned with the ECBs found in the five studied species and interestingly 66.3% ECBs colocalized with those fragile sites (FS). Overall, this study presents detailed chromosomal maps of one OWM and four NWM species. This data will be helpful to further investigation on chromosome evolution in NWM and hominoids in general and is prerequisite for correct interpretation of future sequencing based genomic studies in those species.
The protective mechanism of Ginkgolides and Ginkgo flavonoids on the TNF-α induced apoptosis of rat hippocampal neurons and its mechanisms in vitro.Thursday, July 21, 2016
Guo M, Suo Y, Gao Q, Du H, Zeng W, Wang Y, Hu X, Jiang X,
Heliyon. Sep-2015
Ginkgolides and Ginkgo flavonoids might protect against apoptosis of hippocampal neurons through inhibiting death receptor pathway or mitochondrial pathway under TNF-α background.
Donepezil prevents RANK-induced bone loss via inhibition of osteoclast differentiation by downregulating acetylcholinesterase.Thursday, July 21, 2016
Sato T, Enoki Y, Sakamoto Y, Yokota K, Okubo M, Matsumoto M, Hayashi N, Usui M, Kokabu S, Mimura T, Nakazato Y, Araki N, Fukuda T, Okazaki Y, Suda T, Takeda S, Yoda T,
Heliyon. Sep-2015
AChE promotes osteoclast differentiation in vitro. Donepezil inhibits osteoclast function in vitro and prevents bone loss by suppressing bone resorption in vivo, suggesting the possibility that donepezil reduces fracture risk in patients with Alzheimer's disease.
Real-time and label free determination of ligand binding-kinetics to primary cancer tissue specimens; a novel tool for the assessment of biomarker targeting.Thursday, July 21, 2016
Clausen TM, Pereira MA, Oo HZ, Resende M, Gustavson T, Mao Y, Sugiura N, Liew J, Fazli L, Theander TG, Daugaard M, Salanti A,
Sensing and Bio-Sensing Research. Jul-2016
In clinical oncology, diagnosis and evaluation of optimal treatment strategies are mostly based on histopathological examination combined with immunohistochemical (IHC) expression analysis of cancer-associated antigens in formalin fixed paraffin-embedded (FFPE) tissue biopsies. However, informative IHC analysis depends on both the specificity and affinity of the binding reagent, which are inherently difficult to quantify in situ. Here we describe a label-free method that allows for the direct and real-time assessment of molecular binding kinetics in situ on FFPE tissue specimens using quartz crystal microbalance (QCM) enabled biosensor technology. We analysed the interaction between the rVAR2 protein and its placental-like chondroitin sulfate (pl-CS) receptor in primary human placenta tissue and in breast and prostate tumour specimens in situ. rVAR2 interacted with FFPE human placenta and cancer tissue with an affinity in the nanomolar range, and showed no detectable interaction with pl-CS negative normal tissue. We further validated the method by including analysis with the androgen receptor N-20 antibody (anti-AR). As the KD value produced by this method is independent of the number of epitopes available, this readout offers a quantitative and unbiased readout for in situ binding-avidity and amount of binding epitopes. In summary, this method adds a new and important dimension to classical IHC-based molecular pathology by adding information about the binding characteristics in biologically relevant conditions. This can potentially be used to select optimal biologics for diagnostic and for therapeutic applications as well as guide the development of novel high affinity binding drugs.
Diffusion tensor studies and voxel-based morphometry of the temporal lobe to determine the cognitive prognosis in cases of Alzheimer's disease and mild cognitive impairment: Do white matter changes precede gray matter changes?Thursday, July 21, 2016
Taoka T, Yasuno F, Morikawa M, Inoue M, Kiuchi K, Kitamura S, Matsuoka K, Kishimoto T, Kichikawa K, Naganawa S,
SpringerPlus. 2016
Diffusion tensor parameters (ADC and FA) of the uncinate fascicles correlated well with cognitive function in the next year and seemed to be feasible for use as biomarkers for predicting the progression of AD. In addition, the white matter changes observed in the ADC seemed to precede changes in the gray matter volume of the parahippocampal gyrus that were represented by z-scores of VBM.
Open source approaches to establishing Roseobacter clade bacteria as synthetic biology chassis for biogeoengineering.Thursday, July 21, 2016
Borg Y, Grigonyte AM, Boeing P, Wolfenden B, Smith P, Beaufoy W, Rose S, Ratisai T, Zaikin A, Nesbeth DN,
PeerJ. 2016
Aim. The nascent field of bio-geoengineering stands to benefit from synthetic biologists' efforts to standardise, and in so doing democratise, biomolecular research methods. Roseobacter clade bacteria comprise 15-20% of oceanic bacterio-plankton communities, making them a prime candidate for establishment of synthetic biology chassis for bio-geoengineering activities such as bioremediation of oceanic waste plastic. Developments such as the increasing affordability of DNA synthesis and laboratory automation continue to foster the establishment of a global 'do-it-yourself' research community alongside the more traditional arenas of academe and industry. As a collaborative group of citizen, student and professional scientists we sought to test the following hypotheses: (i) that an incubator capable of cultivating bacterial cells can be constructed entirely from non-laboratory items, (ii) that marine bacteria from the Roseobacter clade can be established as a genetically tractable synthetic biology chassis using plasmids conforming to the BioBrick(TM) standard and finally, (iii) that identifying and subcloning genes from a Roseobacter clade species can readily by achieved by citizen scientists using open source cloning and bioinformatic tools. Method. We cultivated three Roseobacter species, Roseobacter denitrificans, Oceanobulbus indolifexand Dinoroseobacter shibae. For each species we measured chloramphenicol sensitivity, viability over 11 weeks of glycerol-based cryopreservation and tested the effectiveness of a series of electroporation and heat shock protocols for transformation using a variety of plasmid types. We also attempted construction of an incubator-shaker device using only publicly available components. Finally, a subgroup comprising citizen scientists designed and attempted a procedure for isolating the cold resistance anf1 gene from Oceanobulbus indolifexcells and subcloning it into a BioBrick(TM) formatted plasmid. Results. All species were stable over 11 weeks of glycerol cryopreservation, sensitive to 17 µg/mL chloramphenicol and resistant to transformation using the conditions and plasmids tested. An incubator-shaker device, 'UCLHack-12' was assembled and used to cultivate sufficient quantity of Oceanobulbus indolifexcells to enable isolation of the anf1 gene and its subcloning into a plasmid to generate the BioBrick(TM) BBa_K729016. Conclusion.The process of 'de-skilling' biomolecular techniques, particularly for relatively under-investigated organisms, is still on-going. However, our successful cell growth and DNA manipulation experiments serve to indicate the types of capabilities that are now available to citizen scientists. Science democratised in this way can make a positive contribution to the debate around the use of bio-geoengineering to address oceanic pollution or climate change.
Mycobacterium tuberculosis Induces Expansion of Foxp3 Positive CD4 T-cells with a Regulatory Profile in Tuberculin Non-sensitized Healthy Subjects: Implications for Effective Immunization against TB.Thursday, July 21, 2016
Hirsch CS, Rojas R, Wu M, Toossi Z,
Journal of clinical & cellular immunology. Jun-2016
Therefore, MTB and its components expand functional iT-reg in human mononuclear cells from MTB non-sensitized subjects. Also, MTB-induced iT-reg expansion depends on mononuclear phagocyte expression of both TGFβ and IDO.
Plant adaptation to drought stress.Thursday, July 21, 2016
Basu S, Ramegowda V, Kumar A, Pereira A,
F1000Research. 2016
Plants in their natural habitats adapt to drought stress in the environment through a variety of mechanisms, ranging from transient responses to low soil moisture to major survival mechanisms of escape by early flowering in absence of seasonal rainfall. However, crop plants selected by humans to yield products such as grain, vegetable, or fruit in favorable environments with high inputs of water and fertilizer are expected to yield an economic product in response to inputs. Crop plants selected for their economic yield need to survive drought stress through mechanisms that maintain crop yield. Studies on model plants for their survival under stress do not, therefore, always translate to yield of crop plants under stress, and different aspects of drought stress response need to be emphasized. The crop plant model rice ( Oryza sativa) is used here as an example to highlight mechanisms and genes for adaptation of crop plants to drought stress.
Thyroxin Is Useful to Improve Sperm Motility.Thursday, July 21, 2016
Mendeluk GR, Rosales M,
International journal of fertility & sterility. 30-06-2016
We propose a new physiological tool to artificially improve insemination. The discussion opens windows to investigate unknown pathways involved in sperm ca- pacitation and gives innovative arguments to better understand infertility mechanisms.
A Review of The Society for Assisted Reproductive Technology Embryo Grading System and Proposed Modification.Thursday, July 21, 2016
Hossain A, Phelps J, Agarwal A, Sanz E, Mahadevan M,
International journal of fertility & sterility. 01-06-2016
The Society for Assisted Reproductive Technology (SART) method of embryo grad- ing is unique, simple, and widely practiced, and its use has been mandatory for SART membership programs since 2010. Developed by SART in 2006, the current embryo grading system categories, "good, fair, and poor," are limited because they do not describe the best 1-2 embryos in the interest of keeping pace with the shift in clinical practice to be more selective and to transfer fewer embryos. This inspired us to conduct a review on the SART embryo grading system. In this retrospective study, the literature on evaluation of human embryo quality in gen- eral, and the SART method of evaluation in particular, were reviewed for the period of 2000 to 2014. A multifaceted search pertaining to methods of embryo grading and trans- fer using a combination of relevant terms [embryo, mammalian, embryo transfer, grade, grading, morphology, biomarkers, SART, and in vitro fertilization (IVF)] was performed. The inclusion and exclusion in this review were dictated by the aim and scope of the study. Two investigators independently assessed the studies and extracted information. A total of 61 articles were reviewed. Very few studies have evaluated the efficacy of the SART embryo grading method. The present study suggests the necessity for revision of the current SART grading system. The system, as it is now, lacks criteria for describing the cohort specific best embryo and thus is of limited use in single embryo transfer. The study foresees heightened descriptive efficiency of the SART system by implementing the proposed changes. Strengths and weaknesses of the SART embryo grading were identified. Ideas for selecting the best cohort-specific embryo have been discussed, which may trigger methodological improvement in SART and other embryo grading systems.
Bio-Conjugated Gold Nanoparticle Based SERS Probe for Ultrasensitive Identification of Mosquito-Borne Viruses Using Raman Fingerprinting.Thursday, July 21, 2016
Paul AM, Fan Z, Sinha SS, Shi Y, Le L, Bai F, Ray PC,
The journal of physical chemistry. C, Nanomaterials and interfaces. 15-Oct-2015
Dengue virus (DENV) and West Nile virus (WNV) are two well-documented mosquito-borne flaviviruses that cause significant health problems worldwide. Driven by this need, we have developed a bio-conjugated gold nanoparticle (AuNP)-based surface enhanced Raman spectroscopy (SERS) probe for the detection of both DENV and WNV. Reported data demonstrate anti-flavivirus 4G2 antibody conjugated gold nanoparticle (GNP) SERS probe can be used as a Raman fingerprint for the ultrasensitive detection of DENV and WNV selectively. Experimental data show that due to the plasmon coupling in nano-assembly, antibody conjugated GNP- based SERS is able to detect as low as 10 plaque-forming units (PFU)/ml of DENV-2 and WNV, which is comparable with the sensitivity of quantitative PCR-based assays. Selectivity of our probe was demonstrated using another mosquito-borne chikungunya virus (CHIKV) as a negative control. Experimental data demonstrate a huge enhancement of SERS intensity is mainly due to the strong electric field enhancement, which has been confirmed by the finite-difference time-domain (FDTD) simulation. Reported FDTD simulation data indicate the SERS enhancement factor can be more than 10(4) times, due to the assembled structure. Reported results suggest that bio-conjugated AuNP-4G2 based SERS probes have great potential to be used to screen viral particles in clinical and research-based laboratories.
Barriers to Obtaining Sera and Tissue Specimens of African-American Women for the Advancement of Cancer Research.Thursday, July 21, 2016
Strissel KJ, Nicholas DA, Castagne-Charlotin M, Ko N, Denis GV,
Clinical medicine insights. Women's health. 2016
African-American women, a historically understudied and underserved group, have increased risk for triple-negative breast cancer and obesity-associated disease. Obesity-associated metabolic diseases share a common link of low grade chronic inflammation, but not all obese women have metabolic disturbances or are inflamed. One goal of our ongoing research is to identify blood biomarkers that can predict increased risk of breast cancer in women who have obesity or metabolic dysfunction. However, vulnerable populations that stand to benefit most from advances in biomedical research are also underrepresented in research studies. The development of effective, novel approaches for cancer prevention and treatment will require significant basic medical research effort to establish the necessary evidence base in multiple populations. Work with vulnerable human subjects at a safety net hospital enabled us to comment on potential obstacles to obtaining serological and tissue specimens from African-American women. Here, we report some unexpected barriers to participation in our ongoing research study that might inform future efforts.
Structural study of the G57W mutant of human gamma-S-crystallin, associated with congenital cataract.Thursday, July 21, 2016
Khan I, Chandani S, Balasubramanian D,
Molecular vision. 2016
The replacement of glycine at position 57 by the tryptophan residue in human γS-crystallin weakens the stability of the mutant molecule and causes the molecule to self-aggregate, thus generating light-scattering particles. This set of changes in the mutant offers a molecular insight into the mechanism of opacification.
Two-photon Fluorescence Anisotropy Microscopy for Imaging and Direct Measurement of Intracellular Drug Target Engagement.Thursday, July 21, 2016
Vinegoni C, Dubach JM, Feruglio PF, Weissleder R,
IEEE journal of selected topics in quantum electronics : a publication of the IEEE Lasers and Electro-optics Society. 14-07-2016
Small molecule therapeutic drugs must reach their intended cellular targets (pharmacokinetics) and engage them to modulate therapeutic effects (pharmacodynamics). These processes are often difficult to measure in vivo due to their complexities and occurrence within single cells. It has been particularly difficult to directly measure cellular drug target binding. Fluorescence polarization is commonly used in pharmacological screening assays to measure drug-protein or protein-protein interactions. We hypothesized that fluorescence polarization imaging could be adapted and used with fluorescently labeled drugs to measure drug target engagement in vivo. Here we summarize recent results using two photon fluorescence anisotropy microscopy. Our imaging technique offers quantitative pharmacological binding information of diverse molecular interactions at the microscopic level, differentiating between bound and unbound states. Used in combination with other recent advances in the development of novel fluorescently labeled drugs, we expect that the described imaging modality will provide a window into the distribution and efficacy of drugs in real time and in vivo at the cellular and subcellular level.
Relation of inflammatory markers with myocardial and microvascular injury in patients with reperfused ST-elevation myocardial infarction.Thursday, July 21, 2016
Reindl M, Reinstadler SJ, Feistritzer HJ, Klug G, Tiller C, Mair J, Mayr A, Jaschke W, Metzler B,
European heart journal. Acute cardiovascular care. 20-Jul-2016
In reperfused STEMI patients, increased levels of hs-CRP, white blood cell count and fibrinogen are associated with decreased left ventricular function and more pronounced myocardial damage at baseline and 4 months after infarction.
The relationship between vaccine-induced antibody capture of infectious virus and infection outcomes following low-dose, repeated rectal challenge with SIVmac251.Thursday, July 21, 2016
Gach JS, Venzon D, Vaccari M, Keele BF, Franchini G, Forthal DN,
Journal of virology. 20-Jul-2016
Vaccines generally prevent viral infections by eliciting antibodies that inhibit virus infectivity. However, antibodies, including those induced by vaccination, have the potential to enhance, rather than prevent infection. We measured the ability of vaccine-induced antibodies to capture infectious simian immunodeficiency virus (SIV) and explored the relationship between virus capture and infection outcomes. We found that capture correlated with the number of SIV variants that established infection, such that animals whose plasma captured more virus were infected with a higher number of unique strains. In addition, animals whose sera had high capture but weak anti-infectivity activity were infected at a higher rate than were animals with low capture and stronger anti-infectivity activity. These results suggest that vaccines that induce antibodies that bind to and capture infectious virus but don't inhibit virus infectivity will not be effective in preventing infection.
Pharmacokinetics and Exposure-Response Relationship of Golimumab in Patients with Moderately-to-Severely Active Ulcerative Colitis: Results from Phase 2/3 PURSUIT Induction and Maintenance Studies.Thursday, July 21, 2016
Adedokun OJ, Xu Z, Marano CW, Strauss R, Zhang H, Johanns J, Zhou H, Davis HM, Reinisch W, Feagan BG, Rutgeerts P, Sandborn WJ,
Journal of Crohn's & colitis. 20-Jul-2016
SGC are approximately dose-proportional, and a positive SGC-efficacy relationship exists during induction/maintenance golimumab treatment of adult UC patients. Optimal SGC targets require validation in prospective studies.
Specific Blood Pressure Targets for Patients With Diabetic Nephropathy?Thursday, July 21, 2016
Grassi G, Mancia G, Nilsson PM,
Diabetes care. Aug-2016
Diabetic nephropathy represents a condition frequently detected in current clinical practice characterized by a very high cardiovascular risk profile. Blood pressure reduction via antihypertension drug treatment represents a therapeutic approach capable of exerting favorable effects on renal and cardiovascular outcomes. The purpose of this article is to review the current literature and results of key clinical trials pertaining to blood pressure goals of antihypertension treatment in these patients. The pros and cons of a less or a more intensive blood pressure goal in diabetic nephropathy will be discussed, with particular emphasis on the cardiovascular and renal effects of each therapeutic strategy.
Clinical Considerations for Use of Initial Combination Therapy in Type 2 Diabetes.Thursday, July 21, 2016
Cahn A, Cefalu WT,
Diabetes care. Aug-2016
Type 2 diabetes is a progressive disorder characterized by increasing hyperglycemia and the need to gradually intensify therapy in order to achieve and maintain glycemic control. Early initiation of combination therapy has been proposed as an approach to achieve glycemic goals earlier and delay the deterioration of glycemic control and with possible better preservation of β-cell function. We discuss in this article the pros and cons of this approach, focusing on individuals with HbA1c at diagnosis of 7.5-9.0%, where difference of opinion still exists on management. Initial combination therapy is proposed to lead to better and faster achievement of glycemic targets versus monotherapy and to impede clinical inertia and may possibly slow the deterioration of β-cell function. However, treating patients with sequential therapy is proposed to allow one to fully assess the efficacy and risk-to-benefit ratio of each drug as it is added. Furthermore, there is no evidence to support that rapid addition and titration of medications according to the glycemic profile achieved are inferior to initial combination therapy if glycemic targets are attained in a timely manner. Initial combination therapy is argued to postpone clinical inertia to the next decision point but does not eliminate it. Additionally, it may have been the agents chosen and not the timing of their initiation that led to improved β-cell function in the studies of initial combination therapy, and there are no data currently comparing use of the same drugs initiated simultaneously or sequentially. Heightened awareness of providers, individualization of therapy and setting, and reaching glycemic targets remain the mainstays of care.
Characterization of Anaplasma marginale subspecies centrale strains using msp1aS genotyping reveals a wildlife reservoir.Thursday, July 21, 2016
Khumalo ZT, Catanese HN, Liesching N, Hove P, Collins NE, Chaisi ME, Gebremedhin AH, Oosthuizen MC, Brayton KA,
Journal of clinical microbiology. 20-Jul-2016
Bovine anaplasmosis caused by the intraerythrocytic rickettsial pathogen Anaplasma marginale is endemic in South Africa. Anaplasma marginale subspecies centrale (A. centrale) also infects cattle, however, it causes a milder form of anaplasmosis and is used as a live vaccine against A. marginale. There has been less interest in the epidemiology of A. centrale, and, as a result, there are few reports detecting natural infections of this organism. When detected in cattle, it is often assumed that it is due to vaccination, and in most cases it is reported as co-infection with A. marginale without characterization of the strain. In this study a total of 380 blood samples from wild ruminant species and cattle collected from Biobanks, National Parks, and other regions of South Africa were used in duplex real-time PCR assays to simultaneously detect A. marginale and A. centrale. PCR results indicated high occurrence of A. centrale infections ranging from 25-100% in National Parks. Samples positive for A. centrale were further characterized using the msp1aS gene, a homolog of msp1α of A. marginale which contains repeats at the 5' end that are useful for genotyping strains. A total of 47 Msp1aS repeats were identified which corresponded to 32 A. centrale genotypes detected in cattle, buffalo and wildebeest. RepeatAnalyzer was used to examine strain diversity. Our results demonstrate a diversity of A. centrale strains from cattle and wildlife hosts from South Africa and indicate the utility of msp1aS as a genotypic marker for A. centrale strain diversity.
Correlated evolution of male and female reproductive traits drive a cascading effect of reinforcement in Drosophila yakuba.Thursday, July 21, 2016
Comeault AA, Venkat A, Matute DR,
Proceedings. Biological sciences / The Royal Society. 27-Jul-2016
Selection against maladaptive hybridization can drive the evolution of reproductive isolation in a process called reinforcement. While the importance of reinforcement in evolution has been historically debated, many examples now exist. Despite these examples, we typically lack a detailed understanding of the mechanisms limiting the spread of reinforced phenotypes throughout a species' range. Here we address this issue in the fruit fly Drosophila yakuba, a species that hybridizes with its sister species D. santomea and is undergoing reinforcement in a well-defined hybrid zone on the island of São Tomé. Within this region, female D. yakuba show increased postmating-prezygotic (gametic) isolation towards D. santomea when compared with females from allopatric populations. We use a combination of natural collections, fertility assays, and experimental evolution to understand why reinforced gametic isolation in D. yakuba is confined to this hybrid zone. We show that, among other traits, D. yakuba males from sympatric populations sire fewer progeny than allopatric males when mated to allopatric D. yakuba females. Our results provide a novel example of reinforcement acting on a postmating-prezygotic trait in males, resulting in a cascade of reproductive isolation among conspecific populations.
Chemically synthesized peptide libraries as a new source of BBB shuttles. Use of mass spectrometry for peptide identification.Thursday, July 21, 2016
Guixer B, Arroyo X, Belda I, Sabidó E, Teixidó M, Giralt E,
Journal of peptide science : an official publication of the European Peptide Society. 20-Jul-2016
The blood-brain barrier (BBB) is a biological barrier that protects the brain from neurotoxic agents and regulates the influx and efflux of molecules required for its correct function. This stringent regulation hampers the passage of brain parenchyma-targeting drugs across the BBB. BBB shuttles have been proposed as a way to overcome this hurdle because these peptides can not only cross the BBB but also carry molecules which would otherwise be unable to cross the barrier unaided. Here we developed a new high-throughput screening methodology to identify new peptide BBB shuttles in a broadly unexplored chemical space. By introducing d-amino acids, this approach screens only protease-resistant peptides. This methodology combines combinatorial chemistry for peptide library synthesis, in vitro models mimicking the BBB for library evaluation and state-of-the-art mass spectrometry techniques to identify those peptides able to cross the in vitro assays. BBB shuttle synthesis was performed by the mix-and-split technique to generate a library based on the following: Ac-d-Arg-XXXXX-NH2 , where X were: d-Ala (a), d-Arg (r), d-Ile (i), d-Glu (e), d-Ser (s), d-Trp (w) or d-Pro (p). The assays used comprised the in vitro cell-based BBB assay (mimicking both active and passive transport) and the PAMPA (mimicking only passive diffusion). The identification of candidates was determined using a two-step mass spectrometry approach combining LTQ-Orbitrap and Q-trap mass spectrometers. Identified sequences were postulated to cross the BBB models. We hypothesized that some sequences cross the BBB through passive diffusion mechanisms and others through other mechanisms, including paracellular flux and active transport. These results provide a new set of BBB shuttle peptide families. Furthermore, the methodology described is proposed as a consistent approach to search for protease-resistant therapeutic peptides. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.
Optimizing a method for the quantification by quantitative real time PCR of host cell DNA in plasmid vector batches used in human gene therapy.Thursday, July 21, 2016
Ferro S, Fabre I, Chenivesse X,
Human gene therapy methods. 20-Jul-2016
Gene therapy products are very complex advanced therapy medicinal products (ATMP) produced using different processes that require many chemical and biological reagents and production intermediates, such as producing cells. The quantification of residual impurities in gene therapy vectors is a major quality control step when these vectors are used for therapeutic purposes, whether they are derived or not from viruses. Indeed, in non-viral gene therapy products, particularly plasmid vectors used to transfer genetic material, the presence of host-cell DNA (HCDNA) from the bacterial cells used for the vector production is an important concern due to the risk of immunogenicity and insertional mutagenesis. Several methods have been developed to quantify residual HCDNA, but real time quantitative polymerase chain reaction (qPCR) seems to be most suitable because it allows detecting traces of "contaminating" DNA. The French National Agency for Medicines and Health Products Safety (ANSM) ensures the quality and safety of gene transfer medicinal products and must be able to quantify, in its own laboratories, the amount of HCDNA present in plasmid vector batches. Therefore, we developed and validated a qPCR method to quantify at the femtogram level the presence of Escherichia coli residual DNA in plasmid vectors. This approach uses the capillary-based LightCycler® 1.5 System (Roche) with SYBR Green I, a primer pair against the E. coli 23S ribosomal RNA gene and different concentrations of a linearized plasmid that contains the 23S target sequence, as standard. This qPCR method is linear on an 8-decade logarithmic scale, accurate, reproducible and sensitive (quantification of up to 10 copies of 23S target sequence per reaction, or 1.4 E. coli genome, or 7 fg of bacterial DNA). This technique allows ensuring that batches of plasmid vectors to be used in clinical trials comply with the specifications on HCDNA content.
Mid-upper arm circumference is associated with biochemically determined nutritional status indicators among adolescent girls in Central Mozambique.Thursday, July 21, 2016
Kulathinal S, Freese R, Korkalo L, Ismael C, Mutanen M,
Nutrition research (New York, N.Y.). 24-Apr-2016
Biochemically determined nutritional status measurements in low-income countries are often too expensive. Therefore, we hypothesized that some anthropometrical or functional measurements (handgrip) could reflect nutritional status measured by specific biochemical indicators. We did a population-based study from 1 urban area and 2 rural districts in Zambézia Province of Mozambique. The participants (n=386) were non-pregnant adolescent girls between 15 and 18 years of age. 96% had a normal BMI-for-age score. Weight and mid-upper arm circumference (MUAC) were highly correlated (r>0.8) with each other and with total body muscle mass, body mass index (BMI), and with waist circumference, as well as with skinfolds (r>0.6). Upper and total arm lengths were correlated (r>0.7) with height and with each other, and right and left handgrip were correlated only with each other, as were triceps and subscapular skinfolds (r>0.7). Serum albumin correlated negatively with waist circumference (P<.001) and positively with MUAC (P=.007). Stepwise regressions showed that waist circumference, MUAC, weight, and handgrip were important nutritional status indicators in the models using hemoglobin, serum albumin, ferritin, zinc, and plasma retinol concentrations as dependent variables. MUAC could be a valuable anthropometric marker of the overall nutritional status of adolescent girls in low-income countries. When nutrition transition proceeds, waist circumference together with MUAC could form tools for the prediction of worsening of nutritional status.
Acute consumption of organic and conventional tropical grape juices (Vitis labrusca L.) increases antioxidants in plasma and erythrocytes, but not glucose and uric acid levels, in healthy individuals.Thursday, July 21, 2016
Toaldo IM, Cruz FA, da Silva EL, Bordignon-Luiz MT,
Nutrition research (New York, N.Y.). 29-Apr-2016
Bioactive polyphenols in grapes are influenced by grape variety and cultivation conditions. The Vitis labrusca L. varieties are cultivated in tropical regions and used for grape juice production. We hypothesized that polyphenols from tropical grape juices would beneficially affect redox homeostasis in humans. Therefore, the effects of acute consumption of organic and conventional grape juices from V labrusca L. on antioxidants biomarkers were investigated in healthy individuals. In a controlled, randomized, crossover, intervention trial, 24 individuals were assigned to drink 400 mL of conventional juice, organic juice, or water. Each intervention was followed by a 14-day washout period. Blood samples were obtained before and 1 hour after acute intake and analyzed for erythrocyte reduced glutathione, serum total antioxidant capacity, antioxidant enzymes in erythrocytes, and glucose and uric acid in serum. The ingestion of both grape juices resulted in elevated levels of reduced glutathione (P< .001) and serum total antioxidant capacity (P< .05) and increased activity of catalase (P< .001), superoxide dismutase (P< .001), and glutathione peroxidase (P< .05) compared with the control intervention, with no significant differences between grape juices (P< .05). The intake of juices did not affect significantly the concentrations of glucose or uric acid. Grape juice polyphenols were associated with increased antioxidants, and the chemical differences between organic and conventional juices were not predictive of the observed responses. The results suggest a bioactive potential of V labrusca L. juices to improve redox homeostasis, which is involved in defense against oxidative stress in humans.
Rational Design of Fluorescent Phthalazinone Derivatives for One- and Two-Photon Imaging.Thursday, July 21, 2016
Yang L, Zhu Y, Shui M, Zhou T, Cai Y, Wang W, Xu F, Niu Y, Wang C, Zhang JL, Xu P, Yuan L, Liang L,
Chemistry (Weinheim an der Bergstrasse, Germany). 21-Jul-2016
Phthalazinone derivatives were designed as optical probes for one- and two-photon fluorescence microscopy imaging. The design strategy involves stepwise extension and modification of pyridazinone by 1) expansion of pyridazinone to phthalazinone, a larger conjugated system, as the electron acceptor, 2) coupling of electron-donating aromatic groups such as N,N-diethylaminophenyl, thienyl, naphthyl, and quinolyl to the phthalazinone, and 3) anchoring of an alkyl chain to the phthalazinone with various terminal substituents such as triphenylphosphonio, morpholino, triethylammonio, N-methylimidazolio, pyrrolidino, and piperidino. Theoretical calculations were utilized to verify the initial design. The desired fluorescent probes were synthesized by two different routes in considerable yields. Twenty-two phthalazinone derivatives were synthesized and their photophysical properties were measured. Selected compounds were applied in cell imaging, and valuable information was obtained. Furthermore, the designed compounds showed excellent performance in two-photon microscopic imaging of mouse brain slices.
Mobility and toxicity of heavy metal(loid)s arising from contaminated wood ash application to a pasture grassland soil.Thursday, July 21, 2016
Mollon LC, Norton GJ, Trakal L, Moreno-Jimenez E, Elouali FZ, Hough RL, Beesley L,
Environmental pollution (Barking, Essex : 1987). 17-Jul-2016
Heavy metal(loid) rich ash (≤10,000 mg kg(-1) total As, Cr, Cu and Zn) originating from the combustion of contaminated wood was subjected to several experimental procedures involving its incorporation into an upland pasture soil. Ash was added to soil that had been prior amended with local cattle manure, replicating practices employed at the farm scale. Metal(loid) concentrations were measured in soil pore water and ryegrass grown on soil/manure plus ash mixtures (0.1-3.0% vol. ash) in a pot experiment; toxicity evaluation was performed on the same pore water samples by means of a bacterial luminescence biosensor assay. Thereafter a sequential extraction procedure was carried out on selected soil, manure and ash mixtures to elucidate the geochemical association of ash derived metal(loid)s with soil constituents. Predictive modelling was applied to selected data from the pot experiment to determine the risk of transfer of As to meat and milk products in cattle grazing pasture amended with ash. The inclusion of manure to soils receiving ash reduced phyto-toxicity and increased ryegrass biomass yields, compared to soil with ash, but without manure. Elevated As and Cu concentrations in pore water and ryegrass tissue resulting from ash additions were reduced furthest by the inclusion of manure due to an increase in their geochemical association with organic matter. Zinc was the only measured metal(loid) to remain uniformly soluble and bioavailable regardless of the addition of ash and manure. Risk modelling on pot experimental data highlighted that an ash addition of >1% (vol.) to this pasture soil could result in As concentrations in milk and meat products exceeding acceptable limits. The results of this study therefore suggest that even singular low doses of ash applied to soil increase the risk of leaching of metal(loid)s and intensify the risk of As transfer in the food chain.
Alcohol Consumption and Cardiac Biomarkers: The Atherosclerosis Risk in Communities (ARIC) Study.Thursday, July 21, 2016
Lazo M, Chen Y, McEvoy JW, Ndumele C, Konety S, Ballantyne CM, Richey Sharrett A, Selvin E,
Clinical chemistry. 20-Jul-2016
In this community-based study of middle-aged adults without a history of cardiovascular disease, moderate drinking was associated with lower concentrations of hs-cTnT, a marker of chronic subclinical myocardial damage, and positively associated with NT-proBNP, a biomarker of cardiac wall stress. Our results suggest that the cardiac effects of alcohol are complex. Cardiac biomarkers may help improve our understanding of the full cardiovascular effects of alcohol consumption.
Serum Cortisol: An Up-To-Date Assessment of Routine Assay Performance.Thursday, July 21, 2016
Hawley JM, Owen LJ, Lockhart SJ, Monaghan PJ, Armston A, Chadwick CA, Wilshaw H, Freire M, Perry L, Keevil BG,
Clinical chemistry. 20-Jul-2016
Despite the clinical importance of serum cortisol, the performance of routine immunoassays remains highly variable. Accurate quantification is compromised by both matrix effects and antibody specificity. Underpinning this study with a cRMP has highlighted the deficiencies in standardization across routine cortisol immunoassays.
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