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Sample records for cells utilizing immunocytochemistry

  1. Digital microfluidic immunocytochemistry in single cells

    PubMed Central

    Ng, Alphonsus H. C.; Dean Chamberlain, M.; Situ, Haozhong; Lee, Victor; Wheeler, Aaron R.

    2015-01-01

    We report a new technique called Digital microfluidic Immunocytochemistry in Single Cells (DISC). DISC automates protocols for cell culture, stimulation and immunocytochemistry, enabling the interrogation of protein phosphorylation on pulsing with stimulus for as little as 3 s. DISC was used to probe the phosphorylation states of platelet-derived growth factor receptor (PDGFR) and the downstream signalling protein, Akt, to evaluate concentration- and time-dependent effects of stimulation. The high time resolution of the technique allowed for surprising new observations—for example, a 10 s pulse stimulus of a low concentration of PDGF is sufficient to cause >30% of adherent fibroblasts to commit to Akt activation. With the ability to quantitatively probe signalling events with high time resolution at the single-cell level, we propose that DISC may be an important new technique for a wide range of applications, especially for screening signalling responses of a heterogeneous cell population. PMID:26104298

  2. Localization of motilin-immunopositive cells in the rat intestine by light microscopic immunocytochemistry.

    PubMed

    Sakai, T; Satoh, M; Koyama, H; Iesaki, K; Umahara, M; Fujikura, K; Itoh, Z

    1994-01-01

    Motilin-immunopositive cells (Mo cells) are known to exist in the upper small intestine of many species including man. However, the possible presence of Mo cells in the rat gastrointestine has remained obscure because antiserum against it raised in rabbit was found not to cross-react with motilin in the rat gastrointestine. The present study was designed to investigate the distribution of Mo cells in the rat gastrointestine by the peroxidase-conjugated second antibody method using newly raised chicken anti-motilin serum (CPV3). This antiserum was suggested to recognize the N-terminal region of the motilin molecule by enzyme-linked immunosorbent assays and immunocytochemical absorption test. Mo cells detected in the rat gastrointestine by immunocytochemistry were found to be distributed in the duodenum (1.5 cells/mm2), jejunum (2.2 cells/mm2), and ileum (0.028 cells/mm2), and no positive cells were found in the gastric body, gastric antrum, cecum, colon, or pancreas. The immunopositive cells in the rat intestine were spindle shaped or polygonal, scattered throughout the epithelium of the villi and crypts, and similar to those commonly observed in the upper small intestine of other species. These results indicate for the first time that motilin-immunopositive cells do exist in the rat intestine.

  3. Methods for simultaneous interphase in situ hybridization and nuclear antigen immunocytochemistry in T47-D cells.

    PubMed

    Mialhe, A; Cassanelli, S; Louis, J; Seigneurin, D

    1996-02-01

    Procedures that combine immunocytochemistry (ICC) and in situ hybridization (ISH) techniques are now used to investigate phenotype/genotype relationships in the same cells. In this report we describe three rapid procedures for simultaneous detection of a nuclear antigen, progesterone receptors (PR), and the centromeric region of chromosome 11 (to which the human PR gene has been assigned) in T47-D cells. Proteins were stained by precipitates of horseradish peroxidase-diaminobenzidine (PO-DAB, brown color), alkaline phosphatase-Fast Red (APase-Fast Red, red color) or alkaline phosphatase-nitroblue tetrazolium-X-phosphate (APase-NBT-X-Phosphate, blue color) respectively. To obtain a suitable contrast for the two labels, we detected DNA on PO-DAB and APase-NBT-X-phosphate-immunostained cells with interphasic fluorescent in situ hybridization (FISH). By contrast, we combined the APase-Fast Red ICC with an immunocytochemical ISH using alkaline phosphatase-NBT-X-phosphate detection. Only the procedure combining APase-NBT-X-phosphate ICC and FISH ensures optimal visualization of both the PR content and the number of chromosome 11. This method easily provides simultaneous localization of DNA and protein targets in the same cells and should be applicable to many other situations. PMID:8609377

  4. Human medullary thyroid carcinoma: cell cultures and xenotransplants in nude mice. Immunocytochemistry and calcitonin secretion.

    PubMed

    Andry, G; Lothaire, P; Vico, P; Dumont, P; Libert, A; Degeyter, M; Larsimont, D; Saigo, P E; Body, J J; Atassi, G

    1989-12-01

    Occult primary and recurrent medullary thyroid carcinomas (MTC) detected only by elevated calcitonin levels in the peripheral blood, generally after pentagastrin-test stimulation, are difficult to localize. Some new imaging procedures with radionuclide tracers or radiolabelled monoclonal antibodies against carcinoembryonic antigen seem to bring some potentially therapeutic benefits. We report our results with cell cultures and xenotransplants of human MTC with the intention of establishing reproducible models in vitro and in vivo. Cell cultures secrete calcitonin at up to 1200 pg/ml for periods ranging from 3 to 13 weeks. Immunocytochemistry detects cytoplasmic granules positive for calcitonin in polygonal epithelioid cells with dendritic processes. Xenotransplants in nude mice fare better in the subcutaneous axilla than in the subrenal capsule assay. In the former location the tumor-take is good and calcitonin is detected in the blood of the tumor-bearing animals, at levels ranging from 286 to more than 20,000 pg/ml. These models would be potentially usable as targets for radionuclide tracers and/or radiolabelled monoclonal antibodies. PMID:2689238

  5. Detection of neuroblastoma cells in CD34+ selected peripheral stem cells using a combination of tyrosine hydroxylase nested RT-PCR and anti-ganglioside GD2 immunocytochemistry.

    PubMed

    Lode, H N; Handgretinger, R; Schuermann, U; Seitz, G; Klingebiel, T; Niethammer, D; Beck, J

    1997-10-01

    A sensitive assay was developed for the detection of neuroblastoma cell contamination in CD34+ selected and unseparated peripheral blood stem cells (PBSC) used for autologous transplantation in stage 4 neuroblastoma patients. Specifically, we established a non-radioactive nested cDNA-PCR (nPCR) for detection of tyrosine hydroxylase (TH) gene expression combined with anti-disialoganglioside GD2 immunocytochemistry with the murine monoclonal antibody (MAb) 14G2a. Sensitivities of TH nPCR determined with a number of neuroblastoma cell lines and PBSCs correlated to cell line dependent basal TH gene expression levels and ranged from 1:10(4) to 1:10(6). The sensitivity obtained by immunocytochemistry was 1:10(5). We observed the highest PBSC contamination rate of 47% (18/38) among 38 PBSC specimens exclusively obtained from stage 4 neuroblastoma patients by using TH nPCR and GD2 immunocytochemistry in combination. Furthermore, a clinically applied purging method, CD34+ selection by immunoabsorption (CD34+ purity 42.4%), was used on 16 PBSCs. 10/16 (63%) preparations were contaminated prior to CD34+ selection and 56% (9/16) remained contaminated. A significant reduction of neuroblastoma cell contamination by CD34+ selection was not detectable, but the absolute amount of re-infused tumour cells was decreased due to 100-fold smaller cell counts of CD34+ selected grafts used for transplantation. 22 PBSC preparations were used for transplantation. A Kaplan-Meier analysis showed an event-free survival probability of 0.56 +/- 0.22 (n = 9) in the group with contaminated PBSCs versus 0.88 +/- 0.12 (n = 8) with no detectable neuroblastoma-cell contamination. Our data suggest that the combined use of TH nPCR and GD2 immunocytochemistry is optimal to detect contamination and monitor purging strategies. PMID:9516847

  6. Immunoelectron microscopy and immunocytochemistry in pathology, with special reference to immunoglobulin-producing cells.

    PubMed

    Gourdin, M F; Reyes, F; Laurent, G; Gorius, J B

    1979-08-01

    The advantage of immunoelectron microscopy (immuno-EM) is that it allows the simultaneous detection of surface and internal cell components. The immunoperoxidase method is often more suitable than immunoferritin. There are no major difficulties in staining surface antigens by immuno-EM, provided sufficient amounts of pure antibodies are available for coupling to peroxidase. Prior fixation of cells, with faxatives that preserve the antigenicity of surface components, avoids ligand-induced alterations of the surface components. It is believed that, unlike the surface, intracellular antigens are difficult to stain by immuno-EM because of the poor penetration of conjugates into fixed cells; thus, various technical approaches have been proposed by workers involved in tissue immuno-EM. In fact, the method that we initially devised for the surface staining of fixed cell suspensions has proved to detect specifically intracellular immunoglobulins in B cells obtained from patients with proliferative diseases. Thus, conjugates do penetrate into fixed cells, although by an unknown mechanism. On this basis, it is possible to study both surface and intracellular immunoglobulins at the EM level and to determine the precise localization synthesis. PMID:383647

  7. Histochemical demonstration of methamphetamine by immunocytochemistry.

    PubMed

    Ishiyama, I; Mukaida, M; Yoshii, T; Suyama, H

    1987-05-01

    A method for the demonstration of methamphetamine (MA) by immunocytochemistry was established. The tissues of intoxicated mice, administered various amounts of MA in single doses of from 0.01 to 1 mg of MA-HCl, were fixed in glutaraldehyde-containing fixatives. Cryostat and paraffin slices gave a positive reaction of MA localization by staining the brain, liver, kidney, lung, stomach, spleen, and so forth, with the aid of the indirect immunoperoxidase technique. Those of animals administered a single dose of 0.1 mg or more (over 3 to 4 mg/kg--the usual dose of MA in acute intoxication death in forensic medicine), in particular, gave a strong strong reaction, so that the diagnosis of MA intoxication can be performed by macroscopic observation of stained slices. The histochemical diagnosis of MA intoxication in clinical toxicology and pathology might be regarded as a useful tool, especially in forensic pathology. The following cells gave a strong positive reaction: nerve cells and myelin sheaths, hepatocytes, epithelial cells of the distal part of the renal tubule and of the collecting tubule, alveolar and bronchial epithelial cells of the lung, chief and parietal cells of the gastric gland, capillaries of the renal glomerulus, macrophages in the blood and tissues, and striated muscle cells including cardiocytes. The morphological evidence of the pharmacodynamics and pharmacokinetics of MA can be determined at the cellular level by immunocytochemistry.

  8. Three Independent Techniques Localize Expression of Transcript afp-11 and Its Bioactive Peptide Products to the Paired AVK Neurons in Ascaris suum: In Situ Hybridization, Immunocytochemistry, and Single Cell Mass Spectrometry

    PubMed Central

    2012-01-01

    We utilized three independent techniques, immunocytochemistry (ICC), single cell mass spectrometry (MS), and in situ hybridization (ISH), to localize neuropeptides and their transcripts in the nervous system of the nematode Ascaris suum. AF11 (SDIGISEPNFLRFa) is an endogenous peptide with potent paralytic effects on A. suum locomotory behavior. A highly specific antibody to AF11 showed robust immunostaining for AF11 in the paired AVK neurons in the ventral ganglion. We traced the processes from the AVK neurons into the ventral nerve cord and identified them as ventral cord interneurons. MS and MS/MS of single dissected AVKs detected AF11, two previously characterized peptides (AF25 and AF26), seven novel sequence-related peptides, including several sharing a PNFLRFamide C-terminus, and peptide NY, a peptide with an unrelated sequence. Also present in a subset of AVKs was AF2, a peptide encoded by the afp-4 transcript. By sequencing the afp-11 transcript, we discovered that it encodes AF11, all the AF11-related peptides detected by MS in AVK, and peptide NY. ISH detected the afp-11 transcript in AVK neurons, consistent with other techniques. ISH did not detect afp-11 in the ALA neuron, although both ICC and MS found AF11 in ca. 30% of ALAs. All 10 AF11-related peptides reduced acetylcholine-induced muscle contraction, but they differed in their rate of reversal of inhibition after removal of the peptide. PMID:23509978

  9. Three independent techniques localize expression of transcript afp-11 and its bioactive peptide products to the paired AVK neurons in Ascaris suum: in situ hybridization, immunocytochemistry, and single cell mass spectrometry.

    PubMed

    Jarecki, Jessica L; Viola, India R; Andersen, Kari M; Miller, Andrew H; Ramaker, Megan A; Vestling, Martha M; Stretton, Antony O

    2013-03-20

    We utilized three independent techniques, immunocytochemistry (ICC), single cell mass spectrometry (MS), and in situ hybridization (ISH), to localize neuropeptides and their transcripts in the nervous system of the nematode Ascaris suum . AF11 (SDIGISEPNFLRFa) is an endogenous peptide with potent paralytic effects on A. suum locomotory behavior. A highly specific antibody to AF11 showed robust immunostaining for AF11 in the paired AVK neurons in the ventral ganglion. We traced the processes from the AVK neurons into the ventral nerve cord and identified them as ventral cord interneurons. MS and MS/MS of single dissected AVKs detected AF11, two previously characterized peptides (AF25 and AF26), seven novel sequence-related peptides, including several sharing a PNFLRFamide C-terminus, and peptide NY, a peptide with an unrelated sequence. Also present in a subset of AVKs was AF2, a peptide encoded by the afp-4 transcript. By sequencing the afp-11 transcript, we discovered that it encodes AF11, all the AF11-related peptides detected by MS in AVK, and peptide NY. ISH detected the afp-11 transcript in AVK neurons, consistent with other techniques. ISH did not detect afp-11 in the ALA neuron, although both ICC and MS found AF11 in ca. 30% of ALAs. All 10 AF11-related peptides reduced acetylcholine-induced muscle contraction, but they differed in their rate of reversal of inhibition after removal of the peptide.

  10. The Relevance of CD117-Immunocytochemistry Staining Patterns to Mutational Exon-11 in c-kit Detected by PCR from Fine-Needle Aspirated Canine Mast Cell Tumor Cells

    PubMed Central

    Sailasuta, A.; Ketpun, D.; Piyaviriyakul, P.; Theerawatanasirikul, S.; Theewasutrakul, P.; Rungsipipat, A.

    2014-01-01

    Canine cutaneous mast cell tumors (MCT) are the lethal skin tumors. The biological behavior of the MCT cells is quite varied and unpredictable. Almost MCT dogs usually require a rapid diagnosis and therapy. However, MCT diagnosis and prognosis are still dependent on histopathology which is rather inconvenient, time-consuming, painful, and harmful for some cases. Indeed, MCT can be easily accessible using fine-needle aspiration (FNA). In this study, our biopsy specimens were classified as low- and high-grade MCT based on the novel 2-tier histopathologic grading system. We have demonstrated the usage of fine-needle aspirated MCT cells (FNA-MCT cells) from these specimens as a primary cell source to study the distribution of CD117-immunocytochemistry (CD117-ICC) staining patterns and the frequency of internal tandem duplication- (ITD-) mutant exon-11 of c-kit. The result has substantially shown that there were three staining patterns identified in the cells. Only paranuclear pattern was significantly increased in the cells from high-grade MCT. Altogether, the ITD-mutant exon-11 was also detectable only in these cells. Therefore, the result has supported our hypothesis that there was an increased opportunity to observe a higher CD117-ICC staining pattern and exon-11 mutation in high-grade MCT; even these two parameters may not precisely indicate a histopathological grade. PMID:24701365

  11. Amiodarone pulmonary toxicity: cytopathology, ultrastructure, and immunocytochemistry.

    PubMed

    Bedrossian, C W; Warren, C J; Ohar, J; Bhan, R

    1997-10-01

    One hundred ninety cardiac patients were prospectively enrolled in an amiodarone protocol. Over a 10-year period, 16 patients developed new or progressive respiratory symptoms while taking amiodarone. These symptoms included dyspnea associated with abnormal chest radiographs or new or worsening abnormalities on pulmonary function testing. Specimens for microscopic examination were obtained by fiberoptic bronchoscopy with transbronchial lung biopsy (TBB), bronchoalveolar lavage (BAL), open lung biopsy (OLB), or autopsy. Large foamy macrophages with characteristic lamellated cytoplasmic inclusions were noted in all specimens, regardless of other evidence of pulmonary toxicity, suggesting that foamy macrophages represent a routine drug effect. Foamy macrophages were not present in BAL specimens from 53 normal controls and were rarely seen in specimens from 27 patients who had other interstitial lung diseases. When present, the foamy macrophages were less prominent than those seen in specimens from patients receiving amiodarone. Fibrosis was noted in 11 of 16 histological specimens, whereas type II-cell-hyperplasia was observed in 7 of the 16 specimens. Four of the 16 patients with respiratory symptoms died, and their autopsy revealed a combination of foamy macrophages with fibrosis and type II cell hyperplasia reflective of amiodarone pulmonary toxicity. Hyperplastic type II cells were not found in the absence of fibrosis. Immunocytochemistry allowed differentiation between foamy macrophages and type II cells and represents a useful tool for future investigations of the pathogenesis of amiodarone-induced pulmonary disease.

  12. Bilateral renal T-cell lymphoma with hepatic infiltration and secondary polycythemia in a dog: Utility of cytology slides

    PubMed Central

    Froment, Rémi; Gara-Boivin, Carolyn

    2015-01-01

    This is a case of bilateral renal T-cell lymphoma associated with secondary erythrocytosis in a dog. This case is distinctive in using clonality combined with immunocytochemistry to support the diagnosis, thus emphasizing the utility of cytology slides when histology is unavailable. This combination may be a unique canine lymphoma entity. PMID:26663927

  13. Association of Myosin Va and Schwann cells-derived RNA in mammal myelinated axons, analyzed by immunocytochemistry and confocal FRET microscopy.

    PubMed

    Canclini, Lucía; Wallrabe, Horst; Di Paolo, Andrés; Kun, Alejandra; Calliari, Aldo; Sotelo-Silveira, José Roberto; Sotelo, José Roberto

    2014-03-15

    Evidence from multiple sources supports the hypothesis that Schwann cells in the peripheral nervous system transfer messenger RNA and ribosomes to the axons they ensheath. Several technical and methodological difficulties exist for investigators to unravel this process in myelinated axons - a complex two-cell unit. We present an experimental design to demonstrate that newly synthesized RNA is transferred from Schwann cells to axons in association with Myosin Va. The use of quantitative confocal FRET microscopy to track newly-synthesized RNA and determine the molecular association with Myosin Va, is described in detail.

  14. Production and characterization of an antibody against the human bone GLA protein (BGP/osteocalcin) propeptide and its use in immunocytochemistry of bone cells.

    PubMed

    Kasai, R; Bianco, P; Robey, P G; Kahn, A J

    1994-06-01

    We have generated and characterized an antibody that recognizes the C-terminal sequence of the propeptide of human bone GLA protein (BGP/osteocalcin)(amino acid -26 to -1, with +1 being the amino terminus of the mature protein). The range of sensitivity of the antibody, as determined by enzyme-linked immunosorbent assay (ELISA), was 0.5-250 ng/ml. The antibody effectively recognized pro-BGP in cell layer extracts of transformed cells (KT-005), but did not recognize mature, propeptide-less BGP in the medium from the same cultures. Strong labelling was obtained using this antibody in immunoperoxidase staining or immunofluorescence of both transformed and normal human bone cells in vitro. Monensin significantly altered the intracellular pattern of labelling in immunofluorescence studies, indicating that the recognized antigen was associated with the cellular secretory pathway. We also obtained a specific and strong staining of cells in tissue sections of human fetal bone. Antibodies against the mature protein strongly stained the mineralization front, but did not stain cells to any appreciable level. Newly embedded osteocytes were the predominant cell type stained in such material, suggesting that they may represent the major of BGP in the intact tissue. These observations indicate that BGP synthesis is a late event in osteoblastic development and that antibodies generated against the propeptide sequence are a potentially powerful tool in the analysis of bone tumors and evaluation of osteoblastic differentiation.

  15. Effects of spaceflight in the adductor longus muscle of rats flown in the Soviet Biosatellite COSMOS 2044. A study employing neural cell adhesion molecule (N-CAM) immunocytochemistry and conventional morphological techniques (light and electron microscopy)

    NASA Technical Reports Server (NTRS)

    D'Amelio, F.; Daunton, N. G.

    1992-01-01

    The effects of spaceflight upon the "slow" muscle adductor longus were examined in rats flown in the Soviet Biosatellite COSMOS 2044. The techniques employed included standard methods for light microscopy, neural cell adhesion molecule (N-CAM) immunocytochemistry and electron microscopy. Light microscopic observations revealed myofiber atrophy and segmental necrosis accompanied by cellular infiltrates composed of macrophages, leukocytes and mononuclear cells. Neural cell adhesion molecule immunoreactivity (N-CAM-IR) was seen on the myofiber surface and in regenerating myofibers. Ultrastructural alterations included Z band streaming, disorganization of myofibrillar architecture, sarcoplasmic degradation, extensive segmental necrosis with apparent preservation of the basement membrane, degenerative phenomena of the capillary endothelium and cellular invasion of necrotic areas. Regenerating myofibers were identified by the presence of increased amounts of ribosomal aggregates and chains of polyribosomes associated with myofilaments. The principal electron microscopic changes of the neuromuscular junctions showed axon terminals with a decrease or absence of synaptic vesicles replaced by microtubules and neurofilaments, degeneration of axon terminals, vacant axonal spaces and changes suggestive of axonal sprouting. The present observations suggest that alterations such as myofibrillar disruption and necrosis, muscle regeneration and denervation and synaptic remodeling at the level of the neuromuscular junction may take place during spaceflight.

  16. Canine bicavitary carcinomatosis with transient needle tract metastasis diagnosed by multiplex immunocytochemistry.

    PubMed

    Moore, A Russell; Coffey, Emily; Leavell, Sarah E; Krafsur, Greta; Duncan, Colleen; Dowers, Kristy; Santangelo, Kelly S

    2016-09-01

    A 6-year-old, male castrated, mixed-breed dog was referred to the James L. Voss Veterinary Teaching Hospital at Colorado State University for bicavitary effusion. On examination, the dog was tachycardic and tachypneic with bilaterally decreased lung sounds. Thoracic and abdominal ultrasonic examination revealed pleural and peritoneal effusions, which were aspirated and submitted for fluid analysis and cytology. Both cavity fluids were classified as exudates with a large population of vacuolated mononuclear cells. Multiplex immunocytochemistry (ICC) for cytokeratin and vimentin demonstrated exclusively cytokeratin expression, indicating these cells were of epithelial origin. A full diagnostic evaluation was performed, including CBC, clinical chemistry, a pet-side test for heartworm disease, ehrlichiosis, Lyme disease, and anaplasmosis, imaging modalities of thorax, abdomen, and heart, urinalysis, and fine-needle aspirations of spleen, liver, and popliteal lymph nodes. The dog was diagnosed with pleural and peritoneal carcinoma with presumed carcinomatosis. A single dose of intracavitary carboplatin was administered before discharge, and over a period of 2 weeks, 5 thoracocenteses were performed. A subcutaneous mass was noted at a thoracocentesis site one week after initial presentation. Cytology of the mass was consistent with carcinoma, and neoplastic seeding of the tumor cells from the thoracocentesis was suspected. The dog was euthanized 15 days after the first visit, and a necropsy was performed. Findings were consistent with carcinomatosis secondary to anaplastic pulmonary carcinoma with transient subcutaneous seeding of neoplastic cells during routine thoracocentesis. This case demonstrates the utility of multiplex ICC in the clinical setting. PMID:27571570

  17. Rapid triple-labeling method combining in situ hybridization and double immunocytochemistry.

    PubMed

    Lopez-Sanchez, Carmen; Garcia-Martinez, Virginio; Lawson, Aaron; Chapman, Susan C; Schoenwolf, Gary C

    2004-06-01

    A new, rapid method is described for combining in situ hybridization and immunocytochemistry to define cell populations and to map three-dimensional movements of groups of labeled cells within developing chick embryos. The method allows fluorescently labeled cells to be followed in living embryos and subsequently detected as a permanent reaction product for detailed three-dimensional analysis by immunocytochemistry in histological serial sections. Cell identity can be ascertained using a specific riboprobe and in situ hybridization. With this approach, the movements of two groups of cells can be mapped simultaneously (using two different fluorescent trackers and, subsequently, two different chromogens for immunocytochemistry) to analyze relative movements within an embryo, and when combined with in situ hybridization with a specific riboprobe for cell identity, allows fate mapping studies to be conducted using molecular criteria, rather than solely at morphological/positional criteria. The improved method enables the investigator to extract substantially more information from individual embryos, maximizing the results obtained from labor-intensive fate mapping studies.

  18. Localization of a phytohormone using immunocytochemistry

    PubMed Central

    1983-01-01

    The localization of cytokinins in corn root tips was investigated using antibodies or antibody fragments directed against dihydrozeatin riboside and labeled with rhodamine or colloidal gold. Roots were sectioned at -30 degrees to -40 degrees for immunofluorescence or freeze-substituted in ethanol or acetone and embedded in plastic for electron microscopy. Meristematic cells surrounding the quiescent center as well as root cap cells were specifically labeled using direct immunofluorescence techniques, whereas cells of the quiescent center did not bind label. Tissue sections treated with colloidal gold-labeled antibody fragments had gold particles widely distributed in the cytoplasm. The results show that the quiescent center is not the major site of cytokinin localization in root tips. PMID:6352710

  19. The Rate of Oxygen Utilization by Cells

    PubMed Central

    Wagner, Brett A.; Venkataraman, Sujatha; Buettner, Garry R.

    2011-01-01

    The discovery of oxygen is considered by some to be the most important scientific discovery of all time – from both physical-chemical/astrophysics and biology/evolution viewpoints. One of the major developments during evolution is the ability to capture dioxygen in the environment and deliver it to each cell in the multicellular, complex mammalian body -- on demand, i.e. just-in-time. Humans use oxygen to extract approximately 2550 Calories (10.4 MJ) from food to meet daily energy requirements. This combustion requires about 22 moles of dioxygen per day, or 2.5 × 10-4 mol s-1. This is an average rate of oxygen utilization of 2.5 × 10-18 mol cell-1 s-1, i.e. 2.5 amol cell-1 s-1. Cells have a wide range of oxygen utilization, depending on cell type, function, and biological status. Measured rates of oxygen utilization by mammalian cells in culture range from <1 to >350 amol cell-1 s-1. There is a loose positive linear correlation of the rate of oxygen consumption (OCR) by mammalian cells in culture with cell volume and cell protein. The use of oxygen by cells and tissues is an essential aspect of the basic redox biology of cells and tissues. This type of quantitative information is fundamental to investigations in quantitative redox biology, especially redox systems biology. PMID:21664270

  20. Gas concentration cells for utilizing energy

    DOEpatents

    Salomon, Robert E.

    1987-01-01

    An apparatus and method for utilizing energy, in which the apparatus may be used for generating electricity or as a heat pump. When used as an electrical generator, two gas concentration cells are connected in a closed gas circuit. The first gas concentration cell is heated and generates electricity. The second gas concentration cell repressurizes the gas which travels between the cells. The electrical energy which is generated by the first cell drives the second cell as well as an electrical load. When used as a heat pump, two gas concentration cells are connected in a closed gas circuit. The first cell is supplied with electrical energy from a direct current source and releases heat. The second cell absorbs heat. The apparatus has no moving parts and thus approximates a heat engine.

  1. Gas concentration cells for utilizing energy

    DOEpatents

    Salomon, R.E.

    1987-06-30

    An apparatus and method are disclosed for utilizing energy, in which the apparatus may be used for generating electricity or as a heat pump. When used as an electrical generator, two gas concentration cells are connected in a closed gas circuit. The first gas concentration cell is heated and generates electricity. The second gas concentration cell repressurizes the gas which travels between the cells. The electrical energy which is generated by the first cell drives the second cell as well as an electrical load. When used as a heat pump, two gas concentration cells are connected in a closed gas circuit. The first cell is supplied with electrical energy from a direct current source and releases heat. The second cell absorbs heat. The apparatus has no moving parts and thus approximates a heat engine. 4 figs.

  2. T-cell prolymphocytic leukemia (T-PLL) with overlapping cytomorphological features with T-CLL and T-ALL: a case initially diagnosed by fine-needle aspiration cytology and immunocytochemistry.

    PubMed

    Das, Dilip K; Pathan, Shahed K; Joneja, Munish; Al-Musawi, Fatma A; John, Bency; Mirza, Kamran R

    2013-04-01

    T-cell prolymphocytic leukemia (T-PLL) is a very unusual form of chronic lymphoproliferative disorder, which has rarely been diagnosed by fine needle aspiration (FNA) cytology. We report one such case with some overlapping cytomorphological features with chronic lymphocytic leukemia and acute lymphoblastic leukemia. A 91-year-old man presented with generalized lymphadenopathy, pleural effusion, ascites, and an ulcerated growth in rectum. FNA smears from the left cervical lymph node showed a monotonous population of small lymphoid cells having small but distinct nucleoli that was initially diagnosed as chronic lymphocytic leukemia (CLL). Smears from the left axillary lymph node contained both small and medium-sized lymphoid cells with frequent hand-mirror cell appearance, which has been described in acute lymphoblatic leukemia (ALL). Immunocyto/histochemical stainings on smears and cell block preparations of the aspirate showed the following immunophenotype: CD3+, CD4+, CD5+, CD7+, CD8-, CD20-, CD23-, and Tdt-. Total peripheral blood leukocyte count was 26.4 × 10(9) /L and total lymphocyte count, 8.3 × 10(9) /L with predominance of small lymphocytes. T-cell nature of the neoplasm was confirmed by biopsies from the cervical lymph node (T-cell lymphoma), bone marrow (T-cell lymphoid neoplasm/chronic lymphocytic leukemia), and the ulcerated rectal lesion (atypical T-cell lymphoproliferative disorder). The patient developed deep vein thrombosis, heparin-induced thrombocytopenia and bleeding from duodenal ulcer. By the time the reports of all the investigations were ready, the patient succumbed to bronchopneumonia. To the best of our knowledge, this T-CLL/T-PLL which was diagnosed initially by FNA cytology with immunocytochemical support is first of its kind to be reported.

  3. Experiment K-7-18: Effects of Spaceflight in the Muscle Adductor Longus of Rats Flown in the Soviet Biosatellite Cosmos 2044. Part 1; A Study Employing Neural Cell Adhesion Molecules (N-CAM) Immunocytochemistry and Conventional Morphological Techniques (Light and Electron Microscopy)

    NASA Technical Reports Server (NTRS)

    Daunton, N. G.; DAmelio, F.; Wu, L.; Ilyina-Kakueva, E. I.; Krasnov, I. B.; Hyde, T. M.; Sigworth, S. K.

    1994-01-01

    The effects of spaceflight upon the 'slow' muscle adductor longus was examined in rats flown in the Soviet Biosatellite COSMOS 2044. Three groups - synchronous, vivarium and basal served as controls. The techniques employed included standard methods for light microscopy, N-CAM immunocytochemistry and electron microscopy. Light microscopic observations revealed myofiber atrophy, contraction bands and segmental necrosis accompanied by cellular infiltrates composed of macrophages, leucocytes and mononuclear cells. N-CAM immunoreactivity was seen (N-CAM-IR) on the myofiber surface, satellite cells and in regenerating myofibers reminiscent of myotubes. Ultrastructural alterations included Z band streaming, disorganization of myofibrillar architecture, sarcoplasmic degradation, extensive segmental necrosis with preservation of the basement membrane, degenerative phenomena of the capillary endothelium and cellular invasion of necrotic areas. Regenerating myofibers were identified by the presence of increased amounts of ribosomal aggregates and chains of polyribosomes associated with myofilaments that displayed varied distributive patterns. The principal electron microscopic changes of the neuromuscular junctions consisted of a decrease or absence of synaptic vesicles, degeneration of axon terminals, increased number of microtubules, vacant axonal spaces and axonal sprouting. The present observations indicate that major alterations such as myofibrillar disruption and necrosis, muscle regeneration and denervation and synaptic remodeling at the level of the neuromuscular junction may take place during spaceflight.

  4. Fuel cell power system for utility vehicle

    SciTech Connect

    Graham, M.; Barbir, F.; Marken, F.; Nadal, M.

    1996-12-31

    Based on the experience of designing and building the Green Car, a fuel cell/battery hybrid vehicle, and Genesis, a hydrogen/oxygen fuel cell powered transporter, Energy Partners has developed a fuel cell power system for propulsion of an off-road utility vehicle. A 10 kW hydrogen/air fuel cell stack has been developed as a prototype for future mass production. The main features of this stack are discussed in this paper. Design considerations and selection criteria for the main components of the vehicular fuel cell system, such as traction motor, air compressor and compressor motor, hydrogen storage and delivery, water and heat management, power conditioning, and control and monitoring subsystem are discussed in detail.

  5. Fine structure and immunocytochemistry of a new chemosensory system in the Chiton larva (Mollusca: Polyplacophora).

    PubMed

    Haszprunar, Gerhard; Friedrich, Stefan; Wanninger, Andreas; Ruthensteiner, Bernhard

    2002-02-01

    Combined electron microscopy and immunocytochemistry of the larvae of several polyplacophoran species (Chiton olivaceus, Lepidochitona aff. corrugata, Mopalia muscosa) revealed a sensory system new to science, a so-called "ampullary system." The cells of the "ampullary system" are arranged in four symmetrically situated pairs lying dorsolaterally and ventrolaterally in the pretrochal part of the trochophore-like larva and they send axons into the cerebral commissure. They are lost at metamorphosis. The fine structure of these cells strongly resembles that of so-called "ampullary cells" known from various sensory organs of other molluscs, such as the apical complex of gastropod and bivalve larvae, osphradia of vetigastropods, and olfactory organs of cephalopods, and nuchal organs of certain polychaetes. The ampullary cells and their nerves are densely stained by anti-FMRF-amide fluorescence dyes, whereas antiserotonin staining is only weak. While cytological homology of the ampullary cells with those of other organs is probable, the ampullary system as a whole is regarded as a synapomorphy of the Polyplacophora or Chitonida.

  6. [Inhibin B immunocytochemistry for the prognosis assessment of undescended testis damage in children].

    PubMed

    Nicòtina, P A; Arena, F; Romeo, C; Ferlazzo, G; Arena, S; Basile, G; Romeo, G

    2001-01-01

    Inhibin B immunocytochemistry of both the alpha- and beta-subunits was studied in testicular biopsies from 18 prepubertal and postpubertal patients, with unilateral or bilateral cryptorchism. The present investigation was carried out to seek any prognostic significance for the expected fertility of such subjects in adulthood. All samples were also evaluated by histological and morphometric assessments, according a 1-6 grading sy-stem. In this way, the individual testicular changes were scored by quantitating tubular and germ cell hypoplasia, Sertoli cell hyperplasia, and peritubular fibrosis, where present. The results showed that in bilateral maldescended testes an unexpected expression often occurred of inhibin B beta-subunit in Sertoli cells, while inhibin B alpha-subunit there did not, denoting an early developmental arrest of the testis. It co-related with the high grade testicular damages, as a poor predictor of spermatogenesis. Unlike, unilateral retained testes mainly expressed inhibin B alpha-subunit, irrespective of tubular changes. In the latter instance, different pathogenetic factors of imbalanced testicular regulation can be perspected, other than the Inhibin-Activin system.

  7. Proliferation indices and p53-immunocytochemistry in uterine mixed mullerian tumors.

    PubMed

    Nicòtina, P A; Ferlazzo, G; Vincelli, A M

    1997-10-01

    Mixed mullerian tumor (MMT) is a biphasic malignancy of elderly women. It, including both a carcinomatous and a sarcomatous component (CC and SC), is regarded as a female genital tract carcinosarcoma (FGTCS). Since current methods to grade CC and SC are not still univocal, the authors estimate mitotic index (MI) and MIB 1-immunolabeling index (MIB 1-LI) as common prognostic indices for the MMT components. They also compare above prognostic indices with p-53 immunocytochemistry, in MMTs. The present study thus points out that: (a) MI of CC and SC areas is consistent with the respective conventional tumor grades; (b) MI averages of CC are higher than those observed in the SC areas; (c) MI and MIB 1-LI of the CC-tumor cells correlate reciprocally in a very significant fashion; (d) A diffuse strong p53 nuclear immunostaining (> 50% cells) is often patent where the highest MI and MIB 1-LI are found. In conclusion, the authors propose MI and MIB 1-LI as two complementary useful indices to assess prognosis of MMTs. They also suggest p53 nuclear immunolabeling should be regarded as an independent biomarker of unfavourable MMT behaviour.

  8. A novel triple-color detection procedure for brightfield microscopy, combining in situ hybridization with immunocytochemistry.

    PubMed

    Speel, E J; Jansen, M P; Ramaekers, F C; Hopman, A H

    1994-10-01

    We describe a fast light microscopic procedure for the simultaneous enzyme cytochemical detection of three different DNA target sequences in contrasting colors in both interphase and metaphase cell preparations. Chromosome-specific DNA probes labeled with either biotin, digoxygenin, or fluorescein were hybridized as a mixture and detected clearly and accurately by precipitates of the horseradish peroxidase-diaminobenzidine (PO-DAB, brown color), alkaline phosphatase-Fast Red (APase-Fast Red, red color), or horseradish peroxidase-tetramethylbenzidine (PO-TMB, green color) reaction, respectively. The PO-TMB reaction product was stabilized effectively by the addition of sodium tungstate to the reaction mixture, thus making the PO-TMB reaction now generally applicable to in situ hybridization (ISH). To avoid mixing of the precipitates of the two PO reactions used in the triple-color ISH method, the first detected PO activity was always completely inactivated by a mild acid treatment before the second one was applied. Finally, the cell preparations were embedded in a thin protein layer cross-linked by formaldehyde to ensure permanent stabilization of the enzyme reaction products and optimal visualization of color contrast. The triple-color ISH detection procedure could be combined with beta-galactosidase-5-bromo-4-chloro-3-indolyl-beta- D-galactoside (beta-Gal-BCIG) immunocytochemistry (ICC), leading to the simultaneous localization of multiple DNA targets and a protein target in the same cell. The described procedure may therefore be a valuable tool in the areas of cytogenetics, cell biology, and molecular pathology. PMID:7930513

  9. The interest of actin immunocytochemistry in diagnostic histopathology.

    PubMed

    Bussolati, G; Papotti, M; Foschini, M P; Eusebi, V

    1987-01-01

    The immunohistochemical detection of different isoforms of actin provides useful data in the histopathological diagnosis of human tumours. The marked resistance of this protein to fixation and embedding procedures makes it a practical alternative to the ultrastructural search of microfilaments in routine tissue sections. Staining with monoclonal antibodies against alpha smooth muscle actin is helpful in the diagnosis of benign and malignant smooth muscle tumours and allows the differentiation of various types of spindle cell sarcomas. These antibodies are also useful in tracing the myoepithelial cells in normal and various pathological conditions of the breast and salivary glands. Skeletal muscle actin is a reliable marker in the diagnosis of rhabdomyosarcomas. Its use should be combined and complemented with other markers (i.e. desmin; foetal, slow and fast myosins; myoglobin) to monitor the degree of rhabdomyoblastic differentiation of the neoplastic cells in single cases, a parameter of potential prognostic value.

  10. Ultrastructural characteristics and DNA immunocytochemistry in human immunodeficiency virus and zidovudine-associated myopathies.

    PubMed

    Pezeshkpour, G; Illa, I; Dalakas, M C

    1991-12-01

    Electron microscopic features of muscle biopsies from 13 human immunodeficiency (HIV)-positive patients who had myopathy while receiving zidovudine (AZT) were compared with biopsies from five patients with HIV-induced myopathy who were not treated with AZT. All specimens showed disorganization of the myofibrillar structures, along with a varying degree of nemaline (rod) bodies, vacuolization, inflammation, and endothelial tubuloreticular profiles. One untreated and all AZT-treated patients had cytoplasmic bodies, which in the latter were abundant, large, and irregular. Two untreated patients had a peculiar osmiophilic destruction of the muscle fibers, with numerous tubuloreticular profiles in the endothelial cells and brisk inflammation that included lymphoplasmatoid cells. The AZT-treated group had ubiquitous abnormal mitochondria that complemented the presence of ragged red fibers seen by light microscopy. There was subsarcolemmal proliferation of mitochondria, with marked variation in size and shape and proliferation or disorganization of their cristae. Paracrystalline inclusions were seen in one patient. Blind re-examination of the electron micrographs showed abnormal mitochondria that readily distinguished patients with AZT-associated myopathy from those with untreated HIV-induced myopathy. Immunocytochemistry using antibodies to single- and double-stranded DNA revealed severe reduction of mitochondrial DNA compared with the normal nuclear DNA. Although the myopathies associated with HIV and AZT share common myopathologic features, the mitochondrial abnormalities are unique to the AZT-treated patients. Since mitochondrial DNA is specifically reduced, the structural changes noted on electron microscopy are probably associated with mitochondrial dysfunction. Zidovudine, a DNA chain terminator that inhibits the mitochondrial gamma-DNA polymerase, is toxic to muscle mitochondria.

  11. Enhanced methanol utilization in direct methanol fuel cell

    DOEpatents

    Ren, Xiaoming; Gottesfeld, Shimshon

    2001-10-02

    The fuel utilization of a direct methanol fuel cell is enhanced for improved cell efficiency. Distribution plates at the anode and cathode of the fuel cell are configured to distribute reactants vertically and laterally uniformly over a catalyzed membrane surface of the fuel cell. A conductive sheet between the anode distribution plate and the anodic membrane surface forms a mass transport barrier to the methanol fuel that is large relative to a mass transport barrier for a gaseous hydrogen fuel cell. In a preferred embodiment, the distribution plate is a perforated corrugated sheet. The mass transport barrier may be conveniently increased by increasing the thickness of an anode conductive sheet adjacent the membrane surface of the fuel cell.

  12. Fuel utilization and fuel sensitivity of solid oxide fuel cells

    NASA Astrophysics Data System (ADS)

    Huang, Kevin

    2011-03-01

    Fuel utilization and fuel sensitivity are two important process variables widely used in operation of SOFC cells, stacks, and generators. To illustrate the technical values, the definitions of these two variables as well as practical examples are particularly given in this paper. It is explicitly shown that the oxygen-leakage has a substantial effect on the actual fuel utilization, fuel sensitivity and V-I characteristics. An underestimation of the leakage flux could potentially results in overly consuming fuel and oxidizing Ni-based anode. A fuel sensitivity model is also proposed to help extract the leakage flux information from a fuel sensitivity curve. Finally, the "bending-over" phenomenon observed in the low-current range of a V-I curve measured at constant fuel-utilization is quantitatively coupled with leakage flux.

  13. Electric utility acid fuel cell stack technology advancement

    NASA Technical Reports Server (NTRS)

    Congdon, J. V.; Goller, G. J.; Greising, G. J.; Obrien, J. J.; Randall, S. A.; Sandelli, G. J.; Breault, R. D.; Austin, G. W.; Bopse, S.; Coykendall, R. D.

    1984-01-01

    The principal effort under this program was directed at the fuel cell stack technology required to accomplish the initial feasibility demonstrations of increased cell stack operating pressures and temperatures, increased cell active area, incorporation of the ribbed substrate cell configuration at the bove conditions, and the introduction of higher performance electrocatalysts. The program results were successful with the primary accomplishments being: (1) fabrication of 10 sq ft ribbed substrate, cell components including higher performing electrocatalysts; (2) assembly of a 10 sq ft, 30-cell short stack; and (3) initial test of this stack at 120 psia and 405 F. These accomplishments demonstrate the feasibility of fabricating and handling large area cells using materials and processes that are oriented to low cost manufacture. An additional accomplishment under the program was the testing of two 3.7 sq ft short stacks at 12 psia/405 F to 5400 and 4500 hours respectively. These tests demonstrate the durability of the components and the cell stack configuration to a nominal 5000 hours at the higher pressure and temperature condition planned for the next electric utility power plant.

  14. Immunocytochemistry of the neuromuscular systems of Loxosomella vivipara and L. parguerensis (Entoprocta: Loxosomatidae).

    PubMed

    Fuchs, Judith; Bright, Monika; Funch, Peter; Wanninger, Andreas

    2006-07-01

    Little detailed information exists on the anatomy of the nervous system and the musculature of Entoprocta. Herein we describe the distribution of the neurotransmitters RFamide and serotonin as well as the myo-anatomy of adults and asexually produced budding stages of the solitary entoproct species Loxosomella vivipara and L. parguerensis using immunocytochemistry and epifluorescence as well as confocal microscopy. The development of the RFamidergic and serotonergic nervous system starts in early budding stages. In the adults, RFamide is present in the bilateral symmetric cerebral ganglion, a pair of oral nerves that innervate two pairs of nerve cell clusters in the heel of the foot, a pair of aboral nerves, the paired lateral nerves, the calyx nerves, the atrial ring nerve, the tentacle nerves, the stomach nerves, and the rectal nerves. Serotonin is only found in the cerebral ganglion, the oral nerves, and in the tentacle nerves. Some differences in the distribution of both neurotransmitters were found between L. vivipara and L. parguerensis and are most obvious in the differing number of large serotonergic perikarya associated with the oral nerves. Nerves arising from the cerebral ganglion and running in a ventral direction have not been described for Entoprocta before, and the homology of these to the ventral nerve cords of other Spiralia is considered possible. The body musculature of both Loxosomella species comprises longitudinal and diagonal muscles in the foot, the stalk, and the calyx. We found several circular muscles in the calyx. The stalk and parts of the foot and the calyx are surrounded by a fine outer layer of ring muscles. In addition to the congruent details regarding the myo-anatomy of both species, species-specific muscle structures could be revealed. The comparison of our data with recent findings of the myo-anatomy of two Loxosoma species indicates that longitudinal and diagonal body muscles, atrial ring muscles, tentacle muscles, esophageal

  15. Fuel cells for electric utility and transportation applications

    NASA Astrophysics Data System (ADS)

    Srinivasan, S.

    1980-04-01

    The status of fuel cell development is reviewed. For terrestrial electric utility applications, the most promising are phosphoric acid, molten carbonate and solid electrolyte fuel cells. The first will be coupled with a reformer (to convert natural gas, petroleum derived and biomass fuels to hydrogen) while the second and third with a coal gasifier. As ground transportation power sources, the promising systems are phosphoric (or alternate acid) and alkaline electrolyte fuel cells. In the first case, methanol is most attractive while in the second, it will be hydrogen stored as a compressed gas or as a hydride. A technoeconomic assessment of a 'Regenerative Hydrogen-Halogen Energy Storage System' demonstrates the prospects of its use for load leveling when coupled with nuclear, solar or wind power plants.

  16. Fuel Cells Utilizing Oxygen From Air at Low Pressures

    NASA Technical Reports Server (NTRS)

    Cisar, Alan; Boyer, Chris; Greenwald, Charles

    2006-01-01

    A fuel cell stack has been developed to supply power for a high-altitude aircraft with a minimum of air handling. The fuel cell is capable of utilizing oxygen from ambient air at low pressure with no need for compression. For such an application, it is advantageous to take oxygen from the air (in contradistinction to carrying a supply of oxygen onboard), but it is a challenging problem to design a fuel-cell stack of reasonable weight that can generate sufficient power while operating at reduced pressures. The present fuel-cell design is a response to this challenge. The design features a novel bipolar plate structure in combination with a gas-diffusion structure based on a conductive metal core and a carbon gas-diffusion matrix. This combination makes it possible for the flow fields in the stack to have a large open fraction (ratio between open volume and total volume) to permit large volumes of air to flow through with exceptionally low backpressure. Operations at reduced pressure require a corresponding increase in the volume of air that must be handled to deliver the same number of moles of oxygen to the anodes. Moreover, the increase in the open fraction, relative to that of a comparable prior fuel-cell design, reduces the mass of the stack. The fuel cell has been demonstrated to operate at a power density as high as 105 W/cm2 at an air pressure as low as 2 psia (absolute pressure 14 kPa), which is the atmospheric pressure at an altitude of about 50,000 ft ( 15.2 km). The improvements in the design of this fuel cell could be incorporated into designs of other fuel cells to make them lighter in weight and effective at altitudes higher than those of prior designs. Potential commercial applications for these improvements include most applications now under consideration for fuel cells.

  17. Induction of cytochrome P-450 in cultured rat hepatocytes. The heterogeneous localization of specific isoenzymes using immunocytochemistry.

    PubMed

    Bars, R G; Mitchell, A M; Wolf, C R; Elcombe, C R

    1989-08-15

    Primary cultures of rat hepatocytes were exposed to phenobarbitone, clofibric acid, beta-naphthoflavone, isosafrole or dexamethasone for 3 days, and the induction of several cytochrome P-450 isoenzymes was demonstrated by increased catalytic activity, by Western blotting and by immunocytochemistry. The profiles of isoenzymes induced in vitro were compared with those induced in liver microsomes of rats dosed with the same agents. Clofibric acid, an agent which has not been thoroughly investigated previously, was shown to induce both in vivo and in vitro several P-450 isoenzymes normally inducible by phenobarbitone (PB1a, PB3a and PB3b) or steroids (PB2c). Immunocytochemical studies demonstrated that the inducible isoenzymes of cytochrome P-450 are not distributed evenly throughout the hepatocyte population, and increasing concentrations of phenobarbitone or beta-naphthoflavone in the medium results in an increasing proportion of 'induced' cells. However, whereas maximal concentrations of beta-naphthoflavone resulted in virtually all cells containing induced levels of MC1b, a maximal concentration of phenobarbitone resulted in only 30% of the cells containing induced levels of PB3a/PB3b. These results are discussed in relation to the heterogeneous distribution and induction of cytochrome P-450 in the intact liver.

  18. Correlation between SD-OCT, immunocytochemistry and functional findings in an animal model of retinal degeneration

    PubMed Central

    Cuenca, Nicolás; Fernández-Sánchez, Laura; Sauvé, Yves; Segura, Francisco J.; Martínez-Navarrete, Gema; Tamarit, José Manuel; Fuentes-Broto, Lorena; Sanchez-Cano, Ana; Pinilla, Isabel

    2014-01-01

    Purpose: The P23H rhodopsin mutation is an autosomal dominant cause of retinitis pigmentosa (RP). The degeneration can be tracked using different anatomical and functional methods. In our case, we evaluated the anatomical changes using Spectral-Domain Optical Coherence Tomography (SD-OCT) and correlated the findings with retinal thickness values determined by immunocytochemistry.Methods: Pigmented rats heterozygous for the P23H mutation, with ages between P18 and P180 were studied. Function was assessed by means of optomotor testing and ERGs. Retinal thicknesses measurements, autofluorescence and fluorescein angiography were performed using Spectralis OCT. Retinas were studied by means of immunohistochemistry. Results: Between P30 and P180, visual acuity decreased from 0.500 to 0.182 cycles per degree (cyc/deg) and contrast sensitivity decreased from 54.56 to 2.98 for a spatial frequency of 0.089 cyc/deg. Only cone-driven b-wave responses reached developmental maturity. Flicker fusions were also comparable at P29 (42 Hz). Double flash-isolated rod-driven responses were already affected at P29. Photopic responses revealed deterioration after P29.A reduction in retinal thicknesses and morphological modifications were seen in OCT sections. Statistically significant differences were found in all evaluated thicknesses. Autofluorescence was seen in P23H rats as sparse dots. Immunocytochemistry showed a progressive decrease in the outer nuclear layer (ONL), and morphological changes. Although anatomical thickness measures were significantly lower than OCT values, there was a very strong correlation between the values measured by both techniques.Conclusions: In pigmented P23H rats, a progressive deterioration occurs in both retinal function and anatomy. Anatomical changes can be effectively evaluated using SD-OCT and immunocytochemistry, with a good correlation between their values, thus making SD-OCT an important tool for research in retinal degeneration. PMID:25565976

  19. Lung adenocarcinoma and its thyroid metastasis characterized on fine-needle aspirates by cytomorphology, immunocytochemistry, and next-generation sequencing.

    PubMed

    Bellevicine, Claudio; Vigliar, Elena; Malapelle, Umberto; Carelli, Emanuele; Fiorelli, Alfonso; Vicidomini, Giovanni; Cappabianca, Salvatore; Santini, Mario; Troncone, Giancarlo

    2015-07-01

    Lung adenocarcinoma and papillary thyroid carcinoma (PTC) share a number of microscopic and immunophenotypical features. Thus, patients presenting with thyroid and lung synchronous neoplasms may be difficult on fine-needle aspiration (FNA) samples to define the site of origin of the malignancy. In the case reported here, inherent to a 57-years-old man presenting with a right lung mass and a large (44 mm) thyroid nodule, an integrated cytological, immunocytochemical and molecular approach enabled to clarify the primary nature of the neoplasm. FNA cytology showed in both sites papillary structures and nuclear changes reminiscent of PTC. The lung origin of the neoplasm was suggested on cell-block immunocytochemistry showing thyroid transcription factor-1 positive and PAX8 and TGB negative neoplastic cells. Next generation sequencing performed on the Ion Torrent platforms by the Ion Ampliseq Colon and Lung Cancer panel showed a similar genomic profile in both neoplastic sites with a concurrent KRAS G12C mutation. An integrated approach on FNA biospecimen is safe, cost effective, and may be coupled effectively with modern ancillary molecular techniques that may be useful, besides their predictive value, as a adjunctive diagnostic tool when the synchronous occurrence of lesions featuring overlapping morphologies challenge the cytopathologist.

  20. Immunocytochemistry of formalin-fixed human brain tissues: microwave irradiation of free-floating sections.

    PubMed

    Shiurba, R A; Spooner, E T; Ishiguro, K; Takahashi, M; Yoshida, R; Wheelock, T R; Imahori, K; Cataldo, A M; Nixon, R A

    1998-01-01

    Formalin fixation, the chemical process in which formaldehyde binds to cells and tissues, is widely used to preserve human brain specimens from autolytic decomposition. Ultrastructure of cellular and mitochondrial membranes is markedly altered by vesiculation, but this does not interfere with diagnostic evaluation of neurohistology by light microscopy. Serious difficulties are encountered, however, when immunocytochemical staining is attempted. Antigens that are immunoreactive in unfixed frozen sections and protein extracts appear to be concealed or destroyed in formalin-fixed tissues. In dilute aqueous solution, formaldehyde is in equilibrium with methylene glycol and its polymeric hydrates, the balance by far in favor of methylene glyco. Carbonylic formaldehyde is a reactive electrophilic species well known for crosslinking functional groups in tissue proteins, nucleic acids, and polysaccharides. Some of its methylene crosslinks are readily hydrolyzed. Others are stable and irreversible. During immunostaining reactions, intra- and inter-molecular links between macromolecules limit antibody permeation of tissue sections, alter protein secondary structure, and reduce accessibility of antigenic determinants . Accordingly, immunoreactivity is diminished for many antigens. Tissues are rapidly penetrated by methylene glycol, but formaldehyde binding to cellular constituents is relatively slow, increasing progressively until equilibrium is reached. In addition, prolonged storage in formalin may result in acidification of human brain specimens. Low pH favors dissociation of methylene glycol into formaldehyde, further reducing both classical staining and antigen detectability. Various procedures have been devised to counter the antigen masking effects of formaldehyde. Examples include pretreatment of tissue sections with proteases, formic acid, or ultrasound. Recently, heating of mounted sections in ionic salt solution by microwave energy was found to restore many

  1. Indirect myosin immunocytochemistry for the identification of fibre types in equine skeletal muscle

    NASA Technical Reports Server (NTRS)

    Sinha, A. K.; Rose, R. J.; Pozgaj, I.; Hoh, J. F.

    1992-01-01

    The histochemical ATPase method for muscle fibre typing was first described by Brooke and Kaiser in 1970. However, problems have been found with the subdivision of type II fibres using this technique. To determine whether indirect myosin immunocytochemistry using anti-slow (5-4D), anti-fast (1A10) and anti-fast red (5-2B) monoclonal antibodies with cross reactivity for type I, II and IIa fibres, respectively, in a number of species, could identify three fibre types in equine skeletal muscle, data on fibre type composition and fibre size obtained using the two different techniques were compared. Results indicate that different myosin heavy chains can coexist in single equine muscle fibres. Type I and type II fibres were identified by immunocytochemistry, but subdivision of type II fibres was not possible. Although the percentage of type I and type II fibres was not significantly different for the two techniques, a few fibres reacted with both the 1A10 and 5-4D antibodies.

  2. The standard scrapie cell assay: development, utility and prospects.

    PubMed

    van der Merwe, Jacques; Aiken, Judd; Westaway, David; McKenzie, Debbie

    2015-01-16

    Prion diseases are a family of fatal neurodegenerative diseases that involve the misfolding of a host protein, PrPC. Measuring prion infectivity is necessary for determining efficacy of a treatment or infectivity of a prion purification procedure; animal bioassays are, however, very expensive and time consuming. The Standard Scrapie Cell Assay (SSCA) provides an alternative approach. The SSCA facilitates quantitative in vitro analysis of prion strains, titres and biological properties. Given its robust nature and potential for high throughput, the SSCA has substantial utility for in vitro characterization of prions and can be deployed in a number of settings. Here we provide an overview on establishing the SSCA, its use in studies of disease dissemination and pathogenesis, potential pitfalls and a number of remaining challenges.

  3. The Standard Scrapie Cell Assay: Development, Utility and Prospects

    PubMed Central

    van der Merwe, Jacques; Aiken, Judd; Westaway, David; McKenzie, Debbie

    2015-01-01

    Prion diseases are a family of fatal neurodegenerative diseases that involve the misfolding of a host protein, PrPC. Measuring prion infectivity is necessary for determining efficacy of a treatment or infectivity of a prion purification procedure; animal bioassays are, however, very expensive and time consuming. The Standard Scrapie Cell Assay (SSCA) provides an alternative approach. The SSCA facilitates quantitative in vitro analysis of prion strains, titres and biological properties. Given its robust nature and potential for high throughput, the SSCA has substantial utility for in vitro characterization of prions and can be deployed in a number of settings. Here we provide an overview on establishing the SSCA, its use in studies of disease dissemination and pathogenesis, potential pitfalls and a number of remaining challenges. PMID:25602372

  4. Biomarker utility of circulating tumor cells in metastatic cutaneous melanoma.

    PubMed

    Khoja, Leila; Lorigan, Paul; Zhou, Cong; Lancashire, Matthew; Booth, Jessica; Cummings, Jeff; Califano, Raffaele; Clack, Glen; Hughes, Andrew; Dive, Caroline

    2013-06-01

    The incidence of melanoma is increasing worldwide. Advances in targeted agents and immunotherapy have improved outcomes in metastatic disease, but biomarkers are required to optimize treatment. We determined the prevalence of circulating tumor cells (CTCs) and explored their utility as prognostic and pharmacodynamic biomarkers. A total of 101 patients with metastatic cutaneous melanoma were recruited prospectively. CTC number was determined using the CellSearch platform and melanoma kits in samples taken at baseline and serially during treatment. CTC numbers ranged between 0 and 36 per 7.5 ml blood; 26% of patients had ≥ 2 CTCs. Baseline CTC number was prognostic for median overall survival (OS) in univariate analysis (2.6 vs. 7.2 months (P<0.011) for patients with ≥ 2 CTCs vs. <2 CTCs, respectively). In multivariate analysis, CTC number was an independent prognostic biomarker of OS (hazard ratio (HR) 2.403, 95% confidence interval (CI) 1.303-4.430, P=0.005). Patients receiving treatment in whom CTC number remained ≥ 2 CTCs during treatment had shorter median OS than those who maintained <2 CTCs (7 vs. 10 months, HR 0.34, 95% CI 0.14-0.81, log-rank test P=0.015). In conclusion, CTC number in metastatic cutaneous melanoma patients is prognostic for OS with a cutoff of 2 CTCs per 7.5 ml blood. CTC number measured before and throughout treatment provided additional prognostic information. Larger studies are warranted to confirm CTC biomarker utility in melanoma patients. PMID:23223143

  5. Direct Carbon Fuel Cell System Utilizing Solid Carbonaceous Fuels

    SciTech Connect

    Turgut Gur

    2010-04-30

    This 1-year project has achieved most of its objective and successfully demonstrated the viability of the fluidized bed direct carbon fuel cell (FB-DCFC) approach under development by Direct Carbon technologies, LLC, that utilizes solid carbonaceous fuels for power generation. This unique electrochemical technology offers high conversion efficiencies, produces proportionately less CO{sub 2} in capture-ready form, and does not consume or require water for gasification. FB-DCFC employs a specialized solid oxide fuel cell (SOFC) arrangement coupled to a Boudouard gasifier where the solid fuel particles are fluidized and reacted by the anode recycle gas CO{sub 2}. The resulting CO is electrochemically oxidized at the anode. Anode supported SOFC structures employed a porous Ni cermet anode layer, a dense yttria stabilized zirconia membrane, and a mixed conducting porous perovskite cathode film. Several kinds of untreated solid fuels (carbon and coal) were tested in bench scale FBDCFC prototypes for electrochemical performance and stability testing. Single cells of tubular geometry with active areas up to 24 cm{sup 2} were fabricated. The cells achieved high power densities up to 450 mW/cm{sup 2} at 850 C using a low sulfur Alaska coal char. This represents the highest power density reported in the open literature for coal based DCFC. Similarly, power densities up to 175 mW/cm{sup 2} at 850 C were demonstrated with carbon. Electrical conversion efficiencies for coal char were experimentally determined to be 48%. Long-term stability of cell performance was measured under galvanostatic conditions for 375 hours in CO with no degradation whatsoever, indicating that carbon deposition (or coking) does not pose any problems. Similar cell stability results were obtained in coal char tested for 24 hours under galvanostatic conditions with no sign of sulfur poisoning. Moreover, a 50-cell planar stack targeted for 1 kW output was fabricated and tested in 95% CO (balance CO{sub 2

  6. Brain c-Fos immunocytochemistry and cytochrome oxidase histochemistry after a fear conditioning task.

    PubMed

    Conejo, Nélida M; González Pardo, Héctor; López, Matías; Cantora, Raúl; Arias, Jorge L

    2007-05-01

    The involvement of the basolateral and the medial amygdala in fear conditioning was evaluated using different markers of neuronal activation. The method described here is a combination of cytochrome oxidase (CO) histochemistry and c-Fos immunocytochemistry on fresh frozen brain sections. Freezing behavior was used as an index of auditory and contextual fear conditioning. As expected, freezing scores were significantly higher in rats exposed to tone-shock pairings in a distinctive environment (conditioned; COND), as compared to rats that did not receive any shocks (UNCD). CO labeling was increased in the basolateral and medial amygdala of the COND group. Conversely, c-Fos expression in the basolateral and medial amygdala was lower in the COND group as compared to the UNCD group. Furthermore, c-Fos expression was particularly high in the medial amygdala of the UNCD group. The data provided by both techniques indicate that these amygdalar nuclei could play different roles on auditory and contextual fear conditioning. PMID:17425902

  7. GROWTH AND METABOLISM OF INDIVIDUAL BACTERIAL CELLS UTILIZING NANOSIMS

    SciTech Connect

    NEALSON, H. K.

    2007-08-03

    This work involved the use of the Nano-SIMS Instrument at Lawrence Livermore Laboratory, in an effort to utilize this unique tool for experiments in Biology. The work consisted primarily of experiments to measure in real time, C and N fixation in cyanobacteria. The work revealed a number of the difficulties in using the nano-SIMS approach with biological material, but with collaboration from a number of individuals at USC and LLNL, major progress was made. The collaborators from LLNL were from the Chemistry Group (Dr. Peter Weber), and the Biology Group (Dr. Jennifer Pett-Ridge). In addition, there were a number of other scientists involved from LLNL. The USC group consisted of Dr. K.H. Nealson, the PI on the grant, Dr. R. Popa, a postdoctoral fellow and research associate at USC, Professor Douglas Capone, and Juliet Finze, a graduate student in biology. Two major experiments were done, both of which yielded new and exciting data. (1) We studied nitrogen and carbon fixation in Anabaena, demonstrating that fixation ofN occurred rapidly in the heterocysts, and that the fixed N was transported rapidly and completely to the vegetative cells. C fixation occurred in the vegetative cells, with labeled C remaining in these cells in support of their growth and metabolism. This work was accepted in the ISME Journal (Nature Publication), and published last month. (2) We studied nitrogen and carbon fixation in Trichodesmium, a non-heterocystous cyanobacterium that also fixes nitrogen. Interestingly, the nitrogen fixation was confined to regions within the filaments that seem to be identical to the so-called cyanophycaen granules. The fixed N is then transported to other parts of the cyanobacterium, as judged by movement of the heavy N throughout the filaments. On the basis of these very exciting results, we have applied for funding from the NSF to continue the collaboration with LLNL. The results of both studies were presented in the summer of 2007 at the Gordon Research

  8. Utilizing FUCCI reporters to understand pluripotent stem cell biology.

    PubMed

    Singh, Amar M; Trost, Robert; Boward, Benjamin; Dalton, Stephen

    2016-05-15

    The fluorescence ubiquitination cell cycle indicator (FUCCI) system provides a powerful method to evaluate cell cycle mechanisms associated with stem cell self-renewal and cell fate specification. By integrating the FUCCI system into human pluripotent stem cells (hPSCs) it is possible to isolate homogeneous fractions of viable cells representative of all cell cycle phases. This method avoids problems associated with traditional tools used for cell cycle analysis such as synchronizing drugs, elutriation and temperature sensitive mutants. Importantly, FUCCI reporters allow cell cycle events in dynamic systems, such as differentiation, to be evaluated. Initial reports on the FUCCI system focused on its strengths in reporting spatio-temporal aspects of cell cycle events in living cells and developmental models. In this report, we describe approaches that broaden the application of FUCCI reporters in PSCs through incorporation of FACS. This approach allows molecular analysis of the cell cycle in stem cell systems that were not previously possible.

  9. Immunocytochemistry versus nucleic acid amplification in fine needle aspirates and tissues of extrapulmonary tuberculosis

    PubMed Central

    Goel, Madhu Mati; Budhwar, Puja; Jain, Amita

    2012-01-01

    Background: Immunocytochemistry (ICC) is an established routine diagnostic adjunct to cytology and histology for tumor diagnosis but has received little attention for diagnosis of tuberculosis. Aims: To have an objective method of direct visualization of mycobacteria or their products in clinical extrapulmonary tuberculosis (EPTB) specimens, immunocytochemical localization of M. tuberculosis antigen by staining with species specific monoclonal antibody to 38-kDa antigen of Mycobacterium tuberculosis complex. Materials and Methods: Immunostaining with specific monoclonal antibody to 38-kDa antigen of Mycobacterium tuberculosis complex was done in fresh and archival fine needle aspirates and tissue granulomata of 302 cases of extrapulmonary tuberculosis and was compared with the molecular diagnostic i.e., nucleic amplification and conventional [Cytomorphology, Ziehl Neelsen (ZN) staining and culture] tests and 386 controls. Results: Diagnostic indices by Bayesian analysis for all types of archival and fresh material varied from 64 to 76% in nucleic acid amplification (NAA) and 96 to 98% in ICC. There was no significant difference in the diagnostic indices of ZN staining and/ or ICC in fresh or archival material whereas the sensitivity of NAA differed significantly in fresh versus archival material both in cytology (71.4% vs 52.1%) and histology (51.1% vs 38.8%). ICC can be easily used on archival smears and formalin-fixed paraffin-embedded tissue sections with almost equal sensitivity and specificity as with fresh material, in contrast to NAA which showed significant difference in test results on archival and fresh material. Conclusions: Low detection sensitivity of MTB DNA in archival material from known tuberculous cases showed the limitation of in-house NAA-based molecular diagnosis. ICC was found to be sensitive, specific and a better technique than NAA and can be used as an adjunct to conventional morphology and ZN staining for the diagnosis of EPTB in tissue

  10. Utilizing Chimeric Antigen Receptors to Direct Natural Killer Cell Activity

    PubMed Central

    Hermanson, David L.; Kaufman, Dan S.

    2015-01-01

    Natural killer (NK) cells represent an attractive lymphocyte population for cancer immunotherapy due to their ability to lyse tumor targets without prior sensitization and without need for human leukocyte antigens-matching. Chimeric antigen receptors (CARs) are able to enhance lymphocyte targeting and activation toward diverse malignancies. CARs consist of an external recognition domain (typically a small chain variable fragment) directed at a specific tumor antigen that is linked with one or more intracellular signaling domains that mediate lymphocyte activation. Most CAR studies have focused on their expression in T cells. However, use of CARs in NK cells is starting to gain traction because they provide a method to redirect these cells more specifically to target refractory cancers. CAR-mediated anti-tumor activity has been demonstrated using NK cell lines, as well as NK cells isolated from peripheral blood, and NK cells produced from human pluripotent stem cells. This review will outline the CAR constructs that have been reported in NK cells with a focus on comparing the use of different signaling domains in combination with other co-activating domains. PMID:25972867

  11. Sendai virus utilizes specific sialyloligosaccharides as host cell receptor determinants.

    PubMed Central

    Markwell, M A; Paulson, J C

    1980-01-01

    Purified sialyltransferases (CMP-N-acetyl-neuraminate:D-galactosyl-glycoprotein N-acetylneuraminyl-transferase, EC 2.4.99.1) in conjunction with neuraminidase (acylneuraminyl hydrolase, EC 3.2.1.18) were used to produce cell surface sialyloligosaccharides of defined sequence to investigate their role in paramyxovirus infection of host cells. Infection of Madin-Darby bovine kidney cells by Sendai virus was monitored by hemagglutination titer of the virus produced and by changes in morphological characteristics. By either criterion, treatment of the cells with Vibrio cholerae neuraminidase to remove cell surface sialic acids rendered them resistant to infection by Sendai virus. Endogenous replacement of receptors by the cell occurred slowly but supported maximal levels of infection within 6 hr. In contrast, sialylation during a 20-min incubation with CMP-sialic acid and beta-galactoside alpha 2,3-sialytransferase restored full susceptibility to infection. This enzyme elaborates the NeuAc alpha 2,3Gal beta 1,3GalNAc (NeuAc, N-acetylneuraminic acid) sequence on glycoproteins and glycolipids. No restoration of infectivity was observed when neuraminidase-treated cells were sialylated by using beta-galactoside alpha 2,6-sialytransferase, which elaborates the NeuAc-alpha 2,6Gal beta 1,4GlcNAc sequence. These results suggest that sialyloligosaccharide receptor determinants of defined sequence are required for Sendai virus infection of host cells. Images PMID:6255459

  12. Electrochemical cell utilizing molten alkali metal electrode-reactant

    DOEpatents

    Virkar, Anil V.; Miller, Gerald R.

    1983-11-04

    An improved electrochemical cell comprising an additive-modified molten alkali metal electrode-reactant and/or electrolyte is disclosed. Various electrochemical cells employing a molten alkali metal, e.g., sodium, electrode in contact with a cationically conductive ceramic membrane experience a lower resistance and a lower temperature coefficient of resistance whenever small amounts of selenium are present at the interface of the electrolyte and the molten alkali metal. Further, cells having small amounts of selenium present at the electrolyte-molten metal interface exhibit less degradation of the electrolyte under long term cycling conditions.

  13. Inverted amorphous silicon solar cell utilizing cermet layers

    DOEpatents

    Hanak, Joseph J.

    1979-01-01

    An amorphous silicon solar cell incorporating a transparent high work function metal cermet incident to solar radiation and a thick film cermet contacting the amorphous silicon opposite to said incident surface.

  14. Microarray Dot Electrodes Utilizing Dielectrophoresis for Cell Characterization

    PubMed Central

    Yafouz, Bashar; Kadri, Nahrizul Adib; Ibrahim, Fatimah

    2013-01-01

    During the last three decades; dielectrophoresis (DEP) has become a vital tool for cell manipulation and characterization due to its non-invasiveness. It is very useful in the trend towards point-of-care systems. Currently, most efforts are focused on using DEP in biomedical applications, such as the spatial manipulation of cells, the selective separation or enrichment of target cells, high-throughput molecular screening, biosensors and immunoassays. A significant amount of research on DEP has produced a wide range of microelectrode configurations. In this paper; we describe the microarray dot electrode, a promising electrode geometry to characterize and manipulate cells via DEP. The advantages offered by this type of microelectrode are also reviewed. The protocol for fabricating planar microelectrodes using photolithography is documented to demonstrate the fast and cost-effective fabrication process. Additionally; different state-of-the-art Lab-on-a-Chip (LOC) devices that have been proposed for DEP applications in the literature are reviewed. We also present our recently designed LOC device, which uses an improved microarray dot electrode configuration to address the challenges facing other devices. This type of LOC system has the capability to boost the implementation of DEP technology in practical settings such as clinical cell sorting, infection diagnosis, and enrichment of particle populations for drug development. PMID:23857266

  15. Direct Utilization of Coal Syngas in High Temperature Fuel Cells

    SciTech Connect

    Celik, Ismail B.

    2014-10-30

    This EPSCoR project had two primary goals: (i) to build infrastructure and work force at WVU to support long-term research in the area of fuel cells and related sciences; (ii) study effects of various impurities found in coal-syngas on performance of Solid Oxide Fuel Cells (SOFC). As detailed in this report the WVU research team has made significant accomplishments in both of these areas. What follows is a brief summary of these accomplishments: State-of-the-art test facilities and diagnostic tools have been built and put into use. These include cell manufacturing, half-cell and full-cell test benches, XPS, XRD, TEM, Raman, EDAX, SEM, EIS, and ESEM equipment, unique in-situ measurement techniques and test benches (Environmental EM, Transient Mass-Spectrometer-MS, and IR Optical Temperature measurements). In addition, computational capabilities have been developed culminating in a multi-scale multi-physics fuel cell simulation code, DREAM-SOFC, as well as a Beowulf cluster with 64 CPU units. We have trained 16 graduate students, 10 postdoctoral fellows, and recruited 4 new young faculty members who have actively participated in the EPSCoR project. All four of these faculty members have already been promoted to the tenured associate professor level. With the help of these faculty and students, we were able to secure 14 research awards/contracts amounting to a total of circa $5.0 Million external funding in closely related areas of research. Using the facilities mentioned above, the effects of PH3, HCl, Cl2, and H2S on cell performance have been studied in detail, mechanisms have been identified, and also remedies have been proposed and demonstrated in the laboratory. For example, it has been determined that PH3 reacts rapidly with Ni to from secondary compounds which may become softer or even melt at high temperature and then induce Ni migration to the surface of the cell changing the material and micro-structural properties of the cell drastically. It is found that

  16. Panel fabrication utilizing GaAs solar cells

    NASA Technical Reports Server (NTRS)

    Mardesich, N.

    1984-01-01

    The development of the GaAs solar cells for space applications is described. The activities in the fabrication of GaAs solar panels are outlined. Panels were fabricated while introducing improved quality control, soldering laydown and testing procedures. These panels include LIPS II, San Marco Satellite, and a low concentration panel for Rockwells' evaluation. The panels and their present status are discussed.

  17. Primary cells utilize halogen-organic charge transfer complex

    NASA Technical Reports Server (NTRS)

    Gutmann, F.; Hermann, A. M.; Rembaum, A.

    1966-01-01

    Electrochemical cells with solid state components employ charge transfer complexes or donor-acceptor complexes in which the donor component is an organic compound and the acceptor component is a halogen. A minor proportion of graphite added to these composition helps reduce the resistivity.

  18. A bioassay for metals utilizing a human cell line.

    PubMed

    Shea, J; Moran, T; Dehn, P F

    2008-06-01

    The purpose of this study was to assess the ability of the HepG2 cell line to function as a bioassay for metal contamination in sediments, using metallothionein (MT) as a biomarker of exposure. Sediments were collected from the eastern and western ends of Lake Erie, extracted using EPA method 200.7, and analyzed for cadmium (Cd), mercury (Hg) and lead (Pb) levels using ICP-AES. Sediment extracts were neutralized then used at a 2.5% final concentration in the exposure medium. MT levels were measured using the cadmium-hemoglobin affinity assay after a 48 h exposure. Fortified blanks from the ICP protocol served as positive controls. Also, HepG2 cells were exposed to Cd, Pb or combinations of Cd and Pb to determine whether or not induction of MT observed in cells exposed to sediment extracts was due to a single metal, combinations of metals, pH, or some other factor. Additionally, cells were exposed to a range of Cd concentrations approximating the levels found in the extracts (0.0005-0.1mg/L) to determine if a concentration-response occurred. Total metal levels ranged from 527 to 33.5mg/kg with lead the predominant metal, accounting for 100-88.9% of the total quantifiable metals in the sediments. The biomarker response (MT induction) was strongly correlated (r2=0.9919, r2=0.990) with total metal and lead levels in the sediments, respectively, which supports recent field studies indicating the biomarker can discern differences in the strength of the inducing agent. Statistically significant MT induction was associated with sediments which contained measurable Cd concentrations and no significant differences were observed when comparing Cd only and Cd+Pb exposed cells indicating no interactions between Cd and Pb were occurring and supporting our finding that Cd was the main inducing agent in sediment extracts. MT levels also increased significantly in a concentration-dependent manner when cells were exposed only to Cd. Results suggest this human bioassay and the MT

  19. Potential benefits and limitations of utilizing chondroprogenitors in cell-based cartilage therapy

    PubMed Central

    Jayasuriya, Chathuraka T.

    2016-01-01

    Chondroprogenitor cells are a subpopulation of multipotent progenitors that are primed for chondrogenesis. They are believed to have the biological repertoire to be ideal for cell-based cartilage therapy. In addition to summarizing recent advances in chondroprogenitor cell characterization, this review discusses the projected pros and cons of utilizing chondroprogenitors in regenerative medicine and compares them to that of preexisting methods, including autologous chondrocyte implantation (ACI) and the utilization of bone marrow derived mesenchymal stem cells (MSCs) for the purpose of cartilage tissue repair. PMID:26075411

  20. Potential benefits and limitations of utilizing chondroprogenitors in cell-based cartilage therapy.

    PubMed

    Jayasuriya, Chathuraka T; Chen, Qian

    2015-01-01

    Chondroprogenitor cells are a subpopulation of multipotent progenitors that are primed for chondrogenesis. They are believed to have the biological repertoire to be ideal for cell-based cartilage therapy. In addition to summarizing recent advances in chondroprogenitor cell characterization, this review discusses the projected pros and cons of utilizing chondroprogenitors in regenerative medicine and compares them with that of pre-existing methods, including autologous chondrocyte implantation (ACI) and the utilization of bone marrow derived mesenchymal stem cells (MSCs) for the purpose of cartilage tissue repair.

  1. Electrochemical cell utilizing selenium as an electrode-reactant

    SciTech Connect

    Virkar, A.V.; Miller, G.R.; Rasmussen, J.R.

    1990-01-23

    This patent describes an electrochemical cell. It comprises: an anolyte containing substantially a molten alkali metal; a solid beta-alumina electrolyte possessing mobile alkali metal ions of the same alkali metal as is present in the anolyte; and a catholyte comprising a mixture of molten selenium and molten sulfur in a molar ration of about 3:1 to about 30:1 selenium to sulfur, wherein at least a portion of the selenium and sulfur is present in elemental form.

  2. Utilizing stem cells for three-dimensional neural tissue engineering.

    PubMed

    Knowlton, Stephanie; Cho, Yongku; Li, Xue-Jun; Khademhosseini, Ali; Tasoglu, Savas

    2016-05-26

    Three-dimensional neural tissue engineering has made great strides in developing neural disease models and replacement tissues for patients. However, the need for biomimetic tissue models and effective patient therapies remains unmet. The recent push to expand 2D neural tissue engineering into the third dimension shows great potential to advance the field. Another area which has much to offer to neural tissue engineering is stem cell research. Stem cells are well known for their self-renewal and differentiation potential and have been shown to give rise to tissues with structural and functional properties mimicking natural organs. Application of these capabilities to 3D neural tissue engineering may be highly useful for basic research on neural tissue structure and function, engineering disease models, designing tissues for drug development, and generating replacement tissues with a patient's genetic makeup. Here, we discuss the vast potential, as well as the current challenges, unique to integration of 3D fabrication strategies and stem cells into neural tissue engineering. We also present some of the most significant recent achievements, including nerve guidance conduits to facilitate better healing of nerve injuries, functional 3D biomimetic neural tissue models, physiologically relevant disease models for research purposes, and rapid and effective screening of potential drugs.

  3. Clinical Utility of Circulating Tumor Cells in ALK-Positive Non-Small-Cell Lung Cancer

    PubMed Central

    Faugeroux, Vincent; Pailler, Emma; Auger, Nathalie; Taylor, Melissa; Farace, Françoise

    2014-01-01

    The advent of rationally targeted therapies such as small-molecule tyrosine kinase inhibitors (TKIs) has considerably transformed the therapeutic management of a subset of patients with non-small-cell lung cancer (NSCLC) harboring defined molecular abnormalities. When such genetic molecular alterations are detected the use of specific TKI has demonstrated better results (overall response rate, progression free survival) compared to systemic therapy. However, the detection of such molecular abnormalities is complicated by the difficulty in obtaining sufficient tumor material, in terms of quantity and quality, from a biopsy. Here, we described how circulating tumor cells (CTCs) can have a clinical utility in anaplastic lymphoma kinase (ALK) positive NSCLC patients to diagnose ALK-EML4 gene rearrangement and to guide therapeutic management of these patients. The ability to detect genetic abnormalities such ALK rearrangement in CTCs shows that these cells could offer new perspectives both for the diagnosis and the monitoring of ALK-positive patients eligible for treatment with ALK inhibitors. PMID:25414829

  4. Clinical Utility of Circulating Tumor Cells in ALK-Positive Non-Small-Cell Lung Cancer.

    PubMed

    Faugeroux, Vincent; Pailler, Emma; Auger, Nathalie; Taylor, Melissa; Farace, Françoise

    2014-01-01

    The advent of rationally targeted therapies such as small-molecule tyrosine kinase inhibitors (TKIs) has considerably transformed the therapeutic management of a subset of patients with non-small-cell lung cancer (NSCLC) harboring defined molecular abnormalities. When such genetic molecular alterations are detected the use of specific TKI has demonstrated better results (overall response rate, progression free survival) compared to systemic therapy. However, the detection of such molecular abnormalities is complicated by the difficulty in obtaining sufficient tumor material, in terms of quantity and quality, from a biopsy. Here, we described how circulating tumor cells (CTCs) can have a clinical utility in anaplastic lymphoma kinase (ALK) positive NSCLC patients to diagnose ALK-EML4 gene rearrangement and to guide therapeutic management of these patients. The ability to detect genetic abnormalities such ALK rearrangement in CTCs shows that these cells could offer new perspectives both for the diagnosis and the monitoring of ALK-positive patients eligible for treatment with ALK inhibitors. PMID:25414829

  5. Application of fuel cells with heat recovery for integrated utility systems

    NASA Technical Reports Server (NTRS)

    Shields, V.; King, J. M., Jr.

    1975-01-01

    This paper presents the results of a study of fuel cell powerplants with heat recovery for use in an integrated utility system. Such a design provides for a low pollution, noise-free, highly efficient integrated utility. Use of the waste heat from the fuel cell powerplant in an integrated utility system for the village center complex of a new community results in a reduction in resource consumption of 42 percent compared to conventional methods. In addition, the system has the potential of operating on fuels produced from waste materials (pyrolysis and digester gases); this would provide further reduction in energy consumption.

  6. Differential Utilization of Dietary Fatty Acids in Benign and Malignant Cells of the Prostate.

    PubMed

    Dueregger, Andrea; Schöpf, Bernd; Eder, Theresa; Höfer, Julia; Gnaiger, Erich; Aufinger, Astrid; Kenner, Lukas; Perktold, Bernhard; Ramoner, Reinhold; Klocker, Helmut; Eder, Iris E

    2015-01-01

    Tumor cells adapt via metabolic reprogramming to meet elevated energy demands due to continuous proliferation, for example by switching to alternative energy sources. Nutrients such as glucose, fatty acids, ketone bodies and amino acids may be utilized as preferred substrates to fulfill increased energy requirements. In this study we investigated the metabolic characteristics of benign and cancer cells of the prostate with respect to their utilization of medium chain (MCTs) and long chain triglycerides (LCTs) under standard and glucose-starved culture conditions by assessing cell viability, glycolytic activity, mitochondrial respiration, the expression of genes encoding key metabolic enzymes as well as mitochondrial mass and mtDNA content. We report that BE prostate cells (RWPE-1) have a higher competence to utilize fatty acids as energy source than PCa cells (LNCaP, ABL, PC3) as shown not only by increased cell viability upon fatty acid supplementation but also by an increased ß-oxidation of fatty acids, although the base-line respiration was 2-fold higher in prostate cancer cells. Moreover, BE RWPE-1 cells were found to compensate for glucose starvation in the presence of fatty acids. Of notice, these findings were confirmed in vivo by showing that PCa tissue has a lower capacity in oxidizing fatty acids than benign prostate. Collectively, these metabolic differences between benign and prostate cancer cells and especially their differential utilization of fatty acids could be exploited to establish novel diagnostic and therapeutic strategies. PMID:26285134

  7. Differential Utilization of Dietary Fatty Acids in Benign and Malignant Cells of the Prostate.

    PubMed

    Dueregger, Andrea; Schöpf, Bernd; Eder, Theresa; Höfer, Julia; Gnaiger, Erich; Aufinger, Astrid; Kenner, Lukas; Perktold, Bernhard; Ramoner, Reinhold; Klocker, Helmut; Eder, Iris E

    2015-01-01

    Tumor cells adapt via metabolic reprogramming to meet elevated energy demands due to continuous proliferation, for example by switching to alternative energy sources. Nutrients such as glucose, fatty acids, ketone bodies and amino acids may be utilized as preferred substrates to fulfill increased energy requirements. In this study we investigated the metabolic characteristics of benign and cancer cells of the prostate with respect to their utilization of medium chain (MCTs) and long chain triglycerides (LCTs) under standard and glucose-starved culture conditions by assessing cell viability, glycolytic activity, mitochondrial respiration, the expression of genes encoding key metabolic enzymes as well as mitochondrial mass and mtDNA content. We report that BE prostate cells (RWPE-1) have a higher competence to utilize fatty acids as energy source than PCa cells (LNCaP, ABL, PC3) as shown not only by increased cell viability upon fatty acid supplementation but also by an increased ß-oxidation of fatty acids, although the base-line respiration was 2-fold higher in prostate cancer cells. Moreover, BE RWPE-1 cells were found to compensate for glucose starvation in the presence of fatty acids. Of notice, these findings were confirmed in vivo by showing that PCa tissue has a lower capacity in oxidizing fatty acids than benign prostate. Collectively, these metabolic differences between benign and prostate cancer cells and especially their differential utilization of fatty acids could be exploited to establish novel diagnostic and therapeutic strategies.

  8. Differential Utilization of Dietary Fatty Acids in Benign and Malignant Cells of the Prostate

    PubMed Central

    Eder, Theresa; Höfer, Julia; Gnaiger, Erich; Aufinger, Astrid; Kenner, Lukas; Perktold, Bernhard; Ramoner, Reinhold; Klocker, Helmut; Eder, Iris E.

    2015-01-01

    Tumor cells adapt via metabolic reprogramming to meet elevated energy demands due to continuous proliferation, for example by switching to alternative energy sources. Nutrients such as glucose, fatty acids, ketone bodies and amino acids may be utilized as preferred substrates to fulfill increased energy requirements. In this study we investigated the metabolic characteristics of benign and cancer cells of the prostate with respect to their utilization of medium chain (MCTs) and long chain triglycerides (LCTs) under standard and glucose-starved culture conditions by assessing cell viability, glycolytic activity, mitochondrial respiration, the expression of genes encoding key metabolic enzymes as well as mitochondrial mass and mtDNA content. We report that BE prostate cells (RWPE-1) have a higher competence to utilize fatty acids as energy source than PCa cells (LNCaP, ABL, PC3) as shown not only by increased cell viability upon fatty acid supplementation but also by an increased ß-oxidation of fatty acids, although the base-line respiration was 2-fold higher in prostate cancer cells. Moreover, BE RWPE-1 cells were found to compensate for glucose starvation in the presence of fatty acids. Of notice, these findings were confirmed in vivo by showing that PCa tissue has a lower capacity in oxidizing fatty acids than benign prostate. Collectively, these metabolic differences between benign and prostate cancer cells and especially their differential utilization of fatty acids could be exploited to establish novel diagnostic and therapeutic strategies. PMID:26285134

  9. Utilizing cell-matrix interactions to modulate gene transfer to stem cells inside hyaluronic acid hydrogels.

    PubMed

    Gojgini, Shiva; Tokatlian, Talar; Segura, Tatiana

    2011-10-01

    The effective delivery of DNA locally would increase the applicability of gene therapy in tissue regeneration, where diseased tissue is to be repaired in situ. One promising approach is to use hydrogel scaffolds to encapsulate and deliver plasmid DNA in the form of nanoparticles to the diseased tissue, so that cells infiltrating the scaffold are transfected to induce regeneration. This study focuses on the design of a DNA nanoparticle-loaded hydrogel scaffold. In particular, this study focuses on understanding how cell-matrix interactions affect gene transfer to adult stem cells cultured inside matrix metalloproteinase (MMP) degradable hyaluronic acid (HA) hydrogel scaffolds. HA was cross-linked to form a hydrogel material using a MMP degradable peptide and Michael addition chemistry. Gene transfer inside these hydrogel materials was assessed as a function of polyplex nitrogen to phosphate ratio (N/P = 5 to 12), matrix stiffness (100-1700 Pa), RGD (Arg-Gly-Asp) concentration (10-400 μM), and RGD presentation (0.2-4.7 RGDs per HA molecule). All variables were found to affect gene transfer to mouse mensenchymal stem cells culture inside the DNA loaded hydrogels. As expected, higher N/P ratios lead to higher gene transfer efficiency but also higher toxicity; softer hydrogels resulted in higher transgene expression than stiffer hydrogels, and an intermediate RGD concentration and RGD clustering resulted in higher transgene expression. We believe that the knowledge gained through this in vitro model can be utilized to design better scaffold-mediated gene delivery for local gene therapy.

  10. The utility of Caco-2 cells in isolation of enteroviruses from environmental and clinical material.

    PubMed

    Wieczorek, Magdalena; Ciaćka, Agnieszka; Witek, Agnieszka; Litwińska, Bogumiła

    2014-01-01

    The work presented here demonstrates the utility of Caco-2 cells in the isolation of enteroviruses (EVs) from environmental and clinical materials. Thirty-two samples of cerebrospinal fluid positive in Pan-entero RT-PCR were taken for EV strain isolation in cell culture. Out of the 32 samples analysed, 22 (68.75%) were positive for enteroviruses by isolation in Caco-2 cells, and 10 (31.25%) were positive by isolation in RD cells. High viral titre in clinical specimens resulted in rate increase for isolation in Caco-2 cells and RD cells (87.5% and 50%, respectively). Also, the probability of isolation of enteroviruses from sewage in Caco-2 cells was 20 times higher that in RD cells. We proved that Caco-2 cells were more effective than RD cells in enterovirus isolation, irrespective of the material used in the inoculation process.

  11. Utility of Thin-Film Solar Cells on Flexible Substrates for Space Power

    NASA Technical Reports Server (NTRS)

    Dickman, J. E.; Hepp, A. F.; Morel, D. L.; Ferekides, C. S.; Tuttle, J. R.; Hoffman, D. J.; Dhere, N. G.

    2004-01-01

    The thin-film solar cell program at NASA GRC is developing solar cell technologies for space applications which address two critical metrics: specific power (power per unit mass) and launch stowed volume. To be competitive for many space applications, an array using thin film solar cells must significantly increase specific power while reducing stowed volume when compared to the present baseline technology utilizing crystalline solar cells. The NASA GRC program is developing two approaches. Since the vast majority of the mass of a thin film solar cell is in the substrate, a thin film solar cell on a very lightweight flexible substrate (polymer or metal films) is being developed as the first approach. The second approach is the development of multijunction thin film solar cells. Total cell efficiency can be increased by stacking multiple cells having bandgaps tuned to convert the spectrum passing through the upper cells to the lower cells. Once developed, the two approaches will be merged to yield a multijunction, thin film solar cell on a very lightweight, flexible substrate. The ultimate utility of such solar cells in space require the development of monolithic interconnections, lightweight array structures, and ultra-lightweight support and deployment techniques.

  12. Utilization and quality of cryopreserved red blood cells in transfusion medicine.

    PubMed

    Henkelman, S; Noorman, F; Badloe, J F; Lagerberg, J W M

    2015-02-01

    Cryopreserved (frozen) red blood cells have been used in transfusion medicine since the Vietnam war. The main method to freeze the red blood cells is by usage of glycerol. Although the usage of cryopreserved red blood cells was promising due to the prolonged storage time and the limited cellular deterioration at subzero temperatures, its usage have been hampered due to the more complex and labour intensive procedure and the limited shelf life of thawed products. Since the FDA approval of a closed (de) glycerolization procedure in 2002, allowing a prolonged postthaw storage of red blood cells up to 21 days at 2-6°C, cryopreserved red blood cells have become a more utilized blood product. Currently, cryopreserved red blood cells are mainly used in military operations and to stock red blood cells with rare phenotypes. Yet, cryopreserved red blood cells could also be useful to replenish temporary blood shortages, to prolong storage time before autologous transfusion and for IgA-deficient patients. This review describes the main methods to cryopreserve red blood cells, explores the quality of this blood product and highlights clinical settings in which cryopreserved red blood cells are or could be utilized. PMID:25471135

  13. Methods for advanced hepatocyte cell culture in microwells utilizing air bubbles.

    PubMed

    Goral, Vasiliy N; Au, Sam H; Faris, Ronald A; Yuen, Po Ki

    2015-02-21

    Flat, two-dimensional (2D) cell culture substrates are simple to use but offer little control over cell morphologies and behavior. In this article, we present a number of novel and unique methods for advanced cell culture in microwells utilizing air bubbles as a way to seed cells in order to provide substantial control over cellular microenvironments and organization to achieve specific cell-based applications. These cell culture methods enable controlled formation of stable air bubbles in the microwells that spontaneously formed when polar solvents such as cell culture media are loaded. The presence of air bubbles (air bubble masking) enables highly controllable cell patterning and organization of seeded cells as well as cell co-culture in microwells. In addition, these cell culture methods are simple to use and implement, yet versatile, and have the potential to provide a wide range of microenvironments to improve in vivo-like behavior for a number of cell types and applications. The air bubble masking technique can also be used to produce a micron thick layer of collagen film suspended on top of the microwells. These collagen film enclosed microwells could provide an easy way for high throughput drug screening and cytotoxicity assays as different drug compounds could be pre-loaded and dried in selected microwells and then released during cell culture.

  14. L-baclofen-sensitive GABAB binding sites in the medial vestibular nucleus localized by immunocytochemistry

    NASA Technical Reports Server (NTRS)

    Holstein, G. R.; Martinelli, G. P.; Cohen, B.

    1992-01-01

    L-Baclofen-sensitive GABAB binding sites in the medial vestibular nucleus (MVN) were identified immunocytochemically and visualized ultrastructurally in L-baclofen-preinjected rats and monkeys, using a mouse monoclonal antibody with specificity for the p-chlorophenyl moiety of baclofen. Saline-preinjected animals showed no immunostain. In drug-injected animals, there was evidence for both pre- and postsynaptic GABAergic inhibition in MVN mediated by GABAB receptors. These neural elements could be utilized in control of velocity storage in the vestibulo-ocular reflex.

  15. Fructose-induced ROS generation impairs glucose utilization in L6 skeletal muscle cells.

    PubMed

    Jaiswal, N; Maurya, C K; Pandey, J; Rai, A K; Tamrakar, A K

    2015-01-01

    High fructose consumption has implicated in insulin resistance and metabolic syndrome. Fructose is a highly lipogenic sugar that has intense metabolic effects in liver. Recent evidences suggest that fructose exposure to other tissues has substantial and profound metabolic consequences predisposing toward chronic conditions such as type 2 diabetes. Since skeletal muscle is the major site for glucose utilization, in the present study we define the effects of fructose exposure on glucose utilization in skeletal muscle cells. Upon fructose exposure, the L6 skeletal muscle cells displayed diminished glucose uptake, glucose transporter type 4 (GLUT4) translocation, and impaired insulin signaling. The exposure to fructose elevated reactive oxygen species (ROS) production in L6 myotubes, accompanied by activation of the stress/inflammation markers c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (ERK1/2), and degradation of inhibitor of NF-κB (IκBα). We found that fructose caused impairment of glucose utilization and insulin signaling through ROS-mediated activation of JNK and ERK1/2 pathways, which was prevented in the presence of antioxidants. In conclusion, our data demonstrate that exposure to fructose induces cell-autonomous oxidative response through ROS production leading to impaired insulin signaling and attenuated glucose utilization in skeletal muscle cells.

  16. P-glycoprotein expression in primary breast cancer detected by immunocytochemistry with two monoclonal antibodies.

    PubMed Central

    Wishart, G. C.; Plumb, J. A.; Going, J. J.; McNicol, A. M.; McArdle, C. S.; Tsuruo, T.; Kaye, S. B.

    1990-01-01

    We have investigated P-glycoprotein (P-gp) expression in samples of primary breast cancer from 29 patients before therapy. We employed immunohistochemical techniques using two monoclonal antibodies (C219 and MRK16) and an indirect alkaline phosphatase method. Heterogeneous expression in epithelial cells was detected with both C219 (21 of 29) and MRK16 (16 of 29). A surprising finding was P-glycoprotein expression in stromal cells with both C219 (26 of 29) and MRK16 (12 of 29). Our results suggest that significant levels of P-glycoprotein expression may be present in breast cancer before exposure to drugs associated with multidrug resistance. Images Figure 1 PMID:1978783

  17. Utilities for High-Throughput Analysis of B-Cell Clonal Lineages

    PubMed Central

    Lees, William D.; Shepherd, Adrian J.

    2015-01-01

    There are at present few tools available to assist with the determination and analysis of B-cell lineage trees from next-generation sequencing data. Here we present two utilities that support automated large-scale analysis and the creation of publication-quality results. The tools are available on the web and are also available for download so that they can be integrated into an automated pipeline. Critically, and in contrast to previously published tools, these utilities can be used with any suitable phylogenetic inference method and with any antibody germline library and hence are species-independent. PMID:26527585

  18. Utilities for High-Throughput Analysis of B-Cell Clonal Lineages.

    PubMed

    Lees, William D; Shepherd, Adrian J

    2015-01-01

    There are at present few tools available to assist with the determination and analysis of B-cell lineage trees from next-generation sequencing data. Here we present two utilities that support automated large-scale analysis and the creation of publication-quality results. The tools are available on the web and are also available for download so that they can be integrated into an automated pipeline. Critically, and in contrast to previously published tools, these utilities can be used with any suitable phylogenetic inference method and with any antibody germline library and hence are species-independent.

  19. Measurement of estrogen receptor status by immunocytochemistry in paraffin wax sections.

    PubMed

    Jasani, Bharat; Douglas-Jones, Anthony; Rhodes, Anthony; Wozniak, Susan; Barrett-Lee, Peter J; Gee, Julie; Nicholson, Robert

    2006-01-01

    The estrogen receptor (ER) status and, to a lesser extent, progesterone receptor status have been recommended by recently published guidelines as important for routine prognostic and predictive evaluation of breast cancer. Although the clinical utility of ER status has been largely validated using biochemical ligand-binding assays such as the dextran-coated, charcoal ligand-binding assay, there has been the need to develop the ER immunocytochemical assay as a more accurate and practical alternative. In particular, ER status as determined on paraffin sections by immunocytochemical assay has been shown to be superior to the ligand-binding assay for predicting response to adjuvant endocrine therapy in breast cancer. The success of the paraffin-section assay is founded on two principles. The first relates to the advent of the heat-mediated, antigen-retrieval technique capable of restoring ER and progesterone receptor antigenicity in routinely prepared diagnostic formalin-fixed, paraffin-embedded tissue sections. The second is associated with the capacity for this substrate to provide more reliable and reproducible semiquantitative assessment of ER status in morphologically better-preserved tissue used routinely for histopathological diagnosis. The aim of this chapter is to describe the methodology currently used for optimal reproducible demonstration, scoring, and assessment of ER status in paraffin wax-embedded tissue sections in relation to the management of breast cancer in a routine or clinical-trial setting.

  20. Enhanced amino acid utilization sustains growth of cells lacking Snf1/AMPK.

    PubMed

    Nicastro, Raffaele; Tripodi, Farida; Guzzi, Cinzia; Reghellin, Veronica; Khoomrung, Sakda; Capusoni, Claudia; Compagno, Concetta; Airoldi, Cristina; Nielsen, Jens; Alberghina, Lilia; Coccetti, Paola

    2015-07-01

    The metabolism of proliferating cells shows common features even in evolutionary distant organisms such as mammals and yeasts, for example the requirement for anabolic processes under tight control of signaling pathways. Analysis of the rewiring of metabolism, which occurs following the dysregulation of signaling pathways, provides new knowledge about the mechanisms underlying cell proliferation. The key energy regulator in yeast Snf1 and its mammalian ortholog AMPK have earlier been shown to have similar functions at glucose limited conditions and here we show that they also have analogies when grown with glucose excess. We show that loss of Snf1 in cells growing in 2% glucose induces an extensive transcriptional reprogramming, enhances glycolytic activity, fatty acid accumulation and reliance on amino acid utilization for growth. Strikingly, we demonstrate that Snf1/AMPK-deficient cells remodel their metabolism fueling mitochondria and show glucose and amino acids addiction, a typical hallmark of cancer cells.

  1. Peptide Ligand Structure and I-Aq Binding Avidity Influence T Cell Signaling Pathway Utilization

    PubMed Central

    Myers, Linda K; Cullins, David L; Park, Jeoung-Eun; Yi, Ae-Kyung; Brand, David D; Rosloniec, Edward F; Stuart, John M; Kang, Andrew H

    2015-01-01

    Factors that drive T cells to signal through differing pathways remain unclear. We have shown that an altered peptide ligand (A9) activates T cells to utilize an alternate signaling pathway which is dependent upon FcRγ and Syk. However, it remains unknown whether the affinity of peptide binding to MHC drives this selection. To answer this question we developed a panel of peptides designed so that amino acids interacting with the p6 and p9 predicted MHC binding pockets were altered. Analogs were tested for binding to I-Aq using a competitive binding assay and selected analogs were administered to arthritic mice. Using the collagen-induced arthritis (CIA) model, arthritis severity was correlated with T cell cytokine production and molecular T cell signaling responses. We establish that reduced affinity of interaction with the MHC correlates with T cell signaling through the alternative pathway, leading ultimately to secretion of suppressive cytokine and attenuation of arthritis. PMID:25982319

  2. Online quantitative phase imaging of vascular endothelial cells under fluid shear stress utilizing digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Odenthal-Schnittler, Maria; Schnittler, Hans Joachim; Kemper, Björn

    2016-03-01

    We have explored the utilization of quantitative phase imaging with digital holographic microscopy (DHM) as a novel tool for quantifying the dynamics of morphologic parameters (morphodynamics) of confluent endothelial cell layers under fluid shear stress conditions. Human umbilical vein endothelial cells (HUVECs) were exposed to fluid shear stress in a transparent cone/plate flow device (BioTech-Flow-System) and imaged with a modular setup for quantitative DHM phase imaging for up to 48 h. The resulting series of quantitative phase image sequences were analyzed for the average surface roughness of the cell layers and cell alignment. Our results demonstrate that quantitative phase imaging is a powerful and reliable tool to quantify the dynamics of morphological adaptation of endothelial cells to fluid shear stress.

  3. Growth factor and small molecule influence on urological tissue regeneration utilizing cell seeded scaffolds.

    PubMed

    Sharma, Arun K; Cheng, Earl Y

    2015-03-01

    Regenerative medicine strategies combine various attributes from multiple disciplines including stem cell biology, chemistry, materials science and medicine. The junction at which these disciplines intersect provides a means to address unmet medical needs in an assortment of pathologies with the goal of creating sustainable, functional replacement tissues. Tissue damage caused by trauma for example, requires rapid responses in order to mitigate further tissue deterioration. Cell/scaffold composites have been utilized to initiate and stabilize regenerative responses in vivo with the hope that functional tissue can be attained. Along with the gross reconfiguration of regenerating tissues, small molecules and growth factors also play a pivotal role in tissue regeneration. Several regenerative studies targeting a variety of urological tissues demonstrate the utility of these small molecules or growth factors in an in vivo setting.

  4. Effect of Substrate and Cell Surface Hydrophobicity on Phosphate Utilization in Bacteria

    PubMed Central

    Lemke, M. J.; Churchill, P. F.; Wetzel, R. G.

    1995-01-01

    We measured the rates of utilization of hydrophobic and hydrophilic phosphate compounds in gram-negative bacteria with different surface hydrophobicities, isolated from wetland habitats. Three hydrophobic and two hydrophilic bacterial species were selected for study by measuring cell adherence to hydrocarbons. The bacteria were grown under phosphorus-limited conditions with P(infi), hydrophilic (beta)-glycerophosphate, or hydrophobic phosphatidic acid as the phosphate source. Hydrophilic bacteria grew most rapidly on P(infi), followed by (beta)-glycerophosphate. Phosphatidic acid did not support growth or did so at a much later time (40 h) than did the other phosphate treatments. Although all hydrophobic species grew well on these substrates, the rate of growth of two Acinetobacter baumannii isolates on phosphatidic acid exceeded the rate of growth on phosphate or (beta)-glycerophosphate. A membrane phospholipid and lipopolysaccharide were used as a source of phosphorus by hydrophobic species, whereas hydrophilic species could not use the membrane phospholipids and used lipopolysaccharide to a lesser extent. Besides hydrophobic interaction between cells and substrate, phosphatase activity, which was cell bound in hydrophilic species but 30 to 50% unbound in hydrophobic species, affected cell growth. Dialyzed culture supernatant containing phosphatase from hydrophobic species increased the phosphate availability to hydrophilic species. Additionally, cellular extracts from a hydrophilic species, when added to hydrophilic cells, permitted growth on hydrophobic phosphate sources. Naturally occurring amphiphilic humic acids affected the utilization of P(infi) and (beta)-glycerophosphate in bacteria with hydrophilic surfaces but did not affect hydrophobic bacteria. Our results indicate that hydrophobic phosphate sources can be used by bacteria isolated from aquatic environments as the sole phosphorus source for growth. This utilization, in part, appears to be related to

  5. Deficit in sustained attention following selective cholinergic lesion of the pedunculopontine tegmental nucleus in rat, as measured with both post-mortem immunocytochemistry and in vivo PET imaging with [¹⁸F]fluoroethoxybenzovesamicol.

    PubMed

    Cyr, Marilyn; Parent, Maxime J; Mechawar, Naguib; Rosa-Neto, Pedro; Soucy, Jean-Paul; Clark, Stewart D; Aghourian, Meghmik; Bedard, Marc-Andre

    2015-02-01

    Cholinergic neurons of the pedunculopontine tegmental nucleus (PPTg) are thought to be involved in cognitive functions such as sustained attention, and lesions of these cells have been documented in patients showing fluctuations of attention such as in Parkinson's disease or dementia with Lewy Body. Animal studies have been conducted to support the role of these cells in attention, but the lesions induced in these animals were not specific to the cholinergic PPTg system, and were assessed by post-mortem methods remotely performed from the in vivo behavioral assessments. Moreover, sustained attention have not been directly assessed in these studies, but rather deduced from indirect measurements. In the present study, rats were assessed on the 5-Choice Serial Reaction Time Task (5-CSRTT), and a specific measure of variability in response latency was created. Animals were observed both before and after selective lesion of the PPTg cholinergic neurons. Brain cholinergic denervation was assessed both in vivo and ex vivo, using PET imaging with [(18)F]fluoroethoxybenzovesamicol ([(18)F]FEOBV) and immunocytochemistry respectively. Results showed that the number of correct responses and variability in response latency in the 5-CSRTT were the only behavioral measures affected following the lesions. These measures were found to correlate significantly with the number of PPTg cholinergic cells, as measured with both [(18)F]FEOBV and immunocytochemistry. This suggests the primary role of the PPTg cholinergic cells in sustained attention. It also allows to reliably use the PET imaging with [(18)F]FEOBV for the purpose of assessing the relationship between behavior and cholinergic innervation in living animals. PMID:25257103

  6. Characteristics of a Nonvolatile SRAM Memory Cell Utilizing a Ferroelectric Transistor

    NASA Technical Reports Server (NTRS)

    Mitchell, Cody; Laws, Crystal; MacLeod, Todd C.; Ho, Fat D.

    2011-01-01

    The SRAM cell circuit is a standard for volatile data storage. When utilizing one or more ferroelectric transistors, the hysteresis characteristics give unique properties to the SRAM circuit, providing for investigation into the development of a nonvolatile memory cell. This paper discusses various formations of the SRAM circuit, using ferroelectric transistors, n-channel and p-channel MOSFETs, and resistive loads. With varied source and supply voltages, the effects on the timing and retention characteristics are investigated, including retention times of up to 24 hours.

  7. Process Developed for Fabricating Engineered Pore Structures for High- Fuel-Utilization Solid Oxide Fuel Cells

    NASA Technical Reports Server (NTRS)

    Sofie, Stephen W.; Cable, Thomas L.; Salamone, Sam M.

    2005-01-01

    Solid oxide fuel cells (SOFCs) have tremendous commercial potential because of their high efficiency, high energy density, and flexible fuel capability (ability to use fossil fuels). The drive for high-power-utilizing, ultrathin electrolytes (less than 10 microns), has placed an increased demand on the anode to provide structural support, yet allow sufficient fuel entry for sustained power generation. Concentration polarization, a condition where the fuel demand exceeds the supply, is evident in all commercial-based anode-supported cells, and it presents a significant roadblock to SOFC commercialization.

  8. Quantitative studies of chicken somatotrophs during growth and development by morphometry, immunocytochemistry, and flow cytometry.

    PubMed

    Malamed, S; Deaver, D; Perez, F; Radecki, S; Gibney, J; Scanes, C G

    1997-10-01

    Changes in the male chicken somatotroph during growth and maturation have been examined by morphometric and immunocytochemical (ICC) analysis of serial sections of the anterior pituitary gland and by flow cytometry of dispersed anterior pituitary cells. ICC showed that somatotrophs are confined to the middle and caudal thirds of the anterior pituitary gland at all ages from 5 to 26 weeks. At a given age somatotrophs are of equal size at all positions along the cephalocaudal axis of the anterior pituitary gland. However, there are age-related changes: from 5 to 11 weeks rises occur in both the mean total somatotroph volume per gland (64%) and the mean number of somatotrophs (78%), while the mean volume of the single somatotroph is unchanged. From 11 to 18 weeks the mean volume of the single somatotroph decreases 41%. From 18 to 26 weeks the mean volume of the somatotroph, the mean total somatotroph volume, and the mean number per gland do not change. Flow cytometry studies suggested that somatotrophs from adults have less growth hormone (GH) than somatotrophs from young birds. The increases in total somatotroph volume and number from 5 to 11 weeks are consistent with the rise in anterior pituitary GH reported previously. Basic quantitative morphological information about age-related changes in somatotrophs is reported here. When combined with additional facts from future work, they may explain the well-documented sharp decline in circulating GH from 5 to 11 weeks.

  9. Utilization of corn cob biochar in a direct carbon fuel cell

    NASA Astrophysics Data System (ADS)

    Yu, Jinshuai; Zhao, Yicheng; Li, Yongdan

    2014-12-01

    Biochar obtained from the pyrolysis of corn cob is used as the fuel of a direct carbon fuel cell (DCFC) employing a composite electrolyte composed of a samarium doped ceria (SDC) and a eutectic carbonate phase. An anode layer made of NiO and SDC is utilized to suppress the cathode corrosion by the molten carbonate and improves the whole cell stability. The anode off-gas of the fuel cell is analyzed with a gas chromatograph. The effect of working temperature on the cell resistance and power output is examined. The maximum power output achieves 185 mW cm-2 at a current density of 340 mA cm-2 and 750 °C. An anode reaction scheme including the Boudouard reaction is proposed.

  10. Cell compressibility studies utilizing noncontact hydrostatic pressure measurements on single living cells in a microchamber

    NASA Astrophysics Data System (ADS)

    Lin, L. A. G.; Liu, A. Q.; Yu, Y. F.; Zhang, C.; Lim, C. S.; Ng, S. H.; Yap, P. H.; Gao, H. J.

    2008-06-01

    A micro-optical-fluidic system (MOFS), which integrates a force generating device and an optical detector, is designed to measure the bulk modulus of a single living cell in real time under a controlled hydrostatic pressure. In this design, the accuracy of the bulk modulus measurement is improved because neither the force generating device nor the optical detector needs to be in contact with the cells. The MOFS device has been used to investigate the mechanotransduction of THP-1 human acute monocytic leukemia cells and the effects of the toxin lipopolysaccharide and colchicine on various properties of these cells.

  11. Rapid immunocytochemistry based on alternating current electric field using squash smear preparation of central nervous system tumors.

    PubMed

    Moriya, Jun; Tanino, Mishie Ann; Takenami, Tomoko; Endoh, Tomoko; Urushido, Masana; Kato, Yasutaka; Wang, Lei; Kimura, Taichi; Tsuda, Masumi; Nishihara, Hiroshi; Tanaka, Shinya

    2016-01-01

    The role of intraoperative pathological diagnosis for central nervous system (CNS) tumors is crucial for neurosurgery when determining the surgical procedure. Especially, treatment of carmustine (BCNU) wafers requires a conclusive diagnosis of high-grade glioma proven by intraoperative diagnosis. Recently, we demonstrated the usefulness of rapid immunohistochemistry (R-IHC) that facilitates antigen-antibody reaction under alternative current (AC) electric field in the intraoperative diagnosis of CNS tumors; however, a higher proportion of water and lipid in the brain parenchyma sometimes leads to freezing artifacts, resulting in poor quality of frozen sections. On the other hand, squash smear preparation of CNS tumors for cytology does not affect the frozen artifacts, and the importance of smear preparation is now being re-recognized as being better than that of the tissue sections. In this study, we established the rapid immunocytochemistry (R-ICC) protocol for squash smears of CNS tumors using AC electric field that takes only 22 min, and demonstrated its usefulness for semi-quantitative Ki-67/MIB-1 labeling index and CD 20 by R-ICC for intraoperative diagnosis. R-ICC by AC electric field may become a substantial tool for compensating R-IHC and will be applied for broad antibodies in the future.

  12. Harvest and utilization of chemical energy in wastes by microbial fuel cells.

    PubMed

    Sun, Min; Zhai, Lin-Feng; Li, Wen-Wei; Yu, Han-Qing

    2016-05-21

    Organic wastes are now increasingly viewed as a resource of energy that can be harvested by suitable biotechnologies. One promising technology is microbial fuel cells (MFC), which can generate electricity from the degradation of organic pollutants. While the environmental benefits of MFC in waste treatment have been recognized, their potential as an energy producer is not fully understood. Although progresses in material and engineering have greatly improved the power output from MFC, how to efficiently utilize the MFC's energy in real-world scenario remains a challenge. In this review, fundamental understandings on the energy-generating capacity of MFC from real waste treatment are provided and the challenges and opportunities are discussed. The limiting factors restricting the energy output and impairing the long-term reliability of MFC are also analyzed. Several energy storage and in situ utilization strategies for the management of MFC's energy are proposed, and future research needs for real-world application of this approach are explored.

  13. Subcutaneous Adipose Tissue–Derived Stem Cell Utility Is Independent of Anatomical Harvest Site

    PubMed Central

    Choudhery, Mahmood S.; Badowski, Michael; Muise, Angela; Pierce, John; Harris, David T.

    2015-01-01

    Abstract One of the challenges for tissue engineering and regenerative medicine is to obtain suitably large cell numbers for therapy. Mesenchymal stem cells (MSCs) can easily be expanded in vitro to obtain large numbers of cells, but this approach may induce cellular senescence. The characteristics of cells are dependent on variables like age, body mass index (BMI), and disease conditions, however, and in the case of adipose tissue–derived stem cells (ASCs), anatomical harvest site is also an important variable that can affect the regenerative potential of isolated cells. We therefore had kept the parameters (age, BMI, disease conditions) constant in this study to specifically assess influence of anatomical sites of individual donors on utility of ASCs. Adipose tissue was obtained from multiple anatomical sites in individual donors, and viability and nucleated cell yield were determined. MSC frequency was enumerated using colony forming unit assay and cells were characterized by flow cytometry. Growth characteristics were determined by long-term population doubling analysis of each sample. Finally, MSCs were induced to undergo adipogenic, osteogenic, and chondrogenic differentiation. To validate the findings, these results were compared with similar single harvest sites from multiple individual patients. The results of the current study indicated that MSCs obtained from multiple harvest sites in a single donor have similar morphology and phenotype. All adipose depots in a single donor exhibited similar MSC yield, viability, frequency, and growth characteristics. Equivalent differentiation capacity into osteocytes, adipocytes, and chondrocytes was also observed. On the basis of results, we conclude that it is acceptable to combine MSCs obtained from various anatomical locations in a single donor to obtain suitably large cell numbers required for therapy, avoiding in vitro senescence and lengthy and expensive in vitro culturing and expansion steps. PMID:26309790

  14. Production of Solar Cells in Space from Non Specific Ores by Utilization of Electronically Enhanced Sputtering

    NASA Technical Reports Server (NTRS)

    Curreri, Peter A.

    2009-01-01

    An ideal method of construction in space would utilize some form of the Universal Differentiator and Universal Constructor as described by Von Neumann (1). The Universal Differentiator is an idealized non ore specific extractive device which is capable of breaking any ore into its constituent elements, and the Universal Constructor can utilize these elements to build any device with controllability to the nanometer scale. During the Human Exploration Initiative program in the early 1990s a conceptual study was done (2) to understand whether such devices were feasible with near term technology for the utilization of space resources and energy. A candidate system was proposed which would utilize electronically enhanced sputtering as the differentiator. Highly ionized ions would be accelerated to a kinetic energy at which the interaction between them and the lattice elections in the ore would be at a maximum. Experiments have shown that the maximum disintegration of raw material occurs at an ion kinetic energy of about 5 MeV, regardless of the composition and structure of the raw material. Devices that could produce charged ion beams in this energy range in space were being tested in the early 1990s. At this energy, for example an ion in a beam of fluorine ions yields about 8 uranium ions from uranium fluoride, 1,400 hydrogen and oxygen atoms from ice, or 7,000 atoms from sulfur dioxide ice. The ions from the disintegrated ore would then be driven by an electrical field into a discriminator in the form of a mass spectrometer, where the magnetic field would divert the ions into collectors for future use or used directly in molecular beam construction techniques. The process would require 10-7 Torr vacuum which would be available in space or on the moon. If the process were used to make thin film silicon solar cells (ignoring any energy inefficiency for beam production), then energy break even for solar cells in space would occur after 14 days.

  15. Bacterial sugar utilization gives rise to distinct single-cell behaviors

    PubMed Central

    Kaznessis, Yiannis; Beisel, Chase L.

    2014-01-01

    Inducible utilization pathways reflect widespread microbial strategies to uptake and consume sugars from the environment. Despite their broad importance and extensive characterization, little is known how these pathways naturally respond to their inducing sugar in individual cells. Here, we performed single-cell analyses to probe the behavior of representative pathways in the model bacterium Escherichia coli. We observed diverse single-cell behaviors, including uniform responses (D-lactose, D-galactose, N-acetylglucosamine, N-acetylneuraminic acid), “all-or-none” responses (D-xylose, L-rhamnose), and complex combinations thereof (L-arabinose, D-gluconate). Mathematical modeling and probing of genetically modified pathways revealed that the simple framework underlying these pathways—inducible transport and inducible catabolism—could give rise to most of these behaviors. Sugar catabolism was also an important feature, as disruption of catabolism eliminated tunable induction as well as enhanced memory of previous conditions. For instance, disruption of catabolism in pathways that respond to endogenously synthesized sugars led to full pathway induction even in the absence of exogenous sugar. Our findings demonstrate the remarkable flexibility of this simple biological framework, with direct implications for environmental adaptation and the engineering of synthetic utilization pathways as titratable expression systems and for metabolic engineering. PMID:24976172

  16. Oncogenic Herpesvirus Utilizes Stress-Induced Cell Cycle Checkpoints for Efficient Lytic Replication.

    PubMed

    Balistreri, Giuseppe; Viiliäinen, Johanna; Turunen, Mikko; Diaz, Raquel; Lyly, Lauri; Pekkonen, Pirita; Rantala, Juha; Ojala, Krista; Sarek, Grzegorz; Teesalu, Mari; Denisova, Oxana; Peltonen, Karita; Julkunen, Ilkka; Varjosalo, Markku; Kainov, Denis; Kallioniemi, Olli; Laiho, Marikki; Taipale, Jussi; Hautaniemi, Sampsa; Ojala, Päivi M

    2016-02-01

    Kaposi's sarcoma herpesvirus (KSHV) causes Kaposi's sarcoma and certain lymphoproliferative malignancies. Latent infection is established in the majority of tumor cells, whereas lytic replication is reactivated in a small fraction of cells, which is important for both virus spread and disease progression. A siRNA screen for novel regulators of KSHV reactivation identified the E3 ubiquitin ligase MDM2 as a negative regulator of viral reactivation. Depletion of MDM2, a repressor of p53, favored efficient activation of the viral lytic transcription program and viral reactivation. During lytic replication cells activated a p53 response, accumulated DNA damage and arrested at G2-phase. Depletion of p21, a p53 target gene, restored cell cycle progression and thereby impaired the virus reactivation cascade delaying the onset of virus replication induced cytopathic effect. Herpesviruses are known to reactivate in response to different kinds of stress, and our study now highlights the molecular events in the stressed host cell that KSHV has evolved to utilize to ensure efficient viral lytic replication.

  17. Diagnostic utility of the cell block method versus the conventional smear study in pleural fluid cytology

    PubMed Central

    Shivakumarswamy, Udasimath; Arakeri, Surekha U; Karigowdar, Mahesh H; Yelikar, BR

    2012-01-01

    Background: The cytological examinations of serous effusions have been well-accepted, and a positive diagnosis is often considered as a definitive diagnosis. It helps in staging, prognosis and management of the patients in malignancies and also gives information about various inflammatory and non-inflammatory lesions. Diagnostic problems arise in everyday practice to differentiate reactive atypical mesothelial cells and malignant cells by the routine conventional smear (CS) method. Aims: To compare the morphological features of the CS method with those of the cell block (CB) method and also to assess the utility and sensitivity of the CB method in the cytodiagnosis of pleural effusions. Materials and Methods: The study was conducted in the cytology section of the Department of Pathology. Sixty pleural fluid samples were subjected to diagnostic evaluation for over a period of 20 months. Along with the conventional smears, cell blocks were prepared by using 10% alcohol–formalin as a fixative agent. Statistical analysis with the ‘z test’ was performed to identify the cellularity, using the CS and CB methods. Mc. Naemer's χ2test was used to identify the additional yield for malignancy by the CB method. Results: Cellularity and additional yield for malignancy was 15% more by the CB method. Conclusions: The CB method provides high cellularity, better architectural patterns, morphological features and an additional yield of malignant cells, and thereby, increases the sensitivity of the cytodiagnosis when compared with the CS method. PMID:22438610

  18. Prognostic utility of routine chimerism testing at 2 to 6 months after allogeneic hematopoietic cell transplantation.

    PubMed

    Mossallam, Ghada I; Kamel, Azza M; Storer, Barry; Martin, Paul J

    2009-03-01

    The utility of routine chimerism analysis as a prognostic indicator of subsequent outcomes after allogeneic hematopoietic cell transplantation (HCT) with myeloablative conditioning regimens remains controversial. To address this controversy, routine chimerism test results at 2 to 6 months after HCT with myeloablative conditioning regimens were evaluated for association with subsequent risk of chronic graft-versus-host disease (GVHD), nonrelapse mortality (NRM), relapse, and overall mortality. Only 70 of 1304 patients (5%) had < 95% donor-derived cells in the marrow. Low donor chimerism in the marrow occurred more often in patients with low-risk diseases compared with those with higher-risk diseases and was significantly associated with a reduced risk of chronic GVHD. Among 673 patients evaluated, 164 (24%) had < 85% donor-derived T cells in the blood. Low donor T cell chimerism was more frequent in patients with low-risk diseases compared with those with higher-risk diseases, in those who received conditioning with busulfan compared with those who received conditioning with total body irradiation, and in those with lower-grade acute GVHD. Low donor T cell chimerism in the blood was significantly associated with a reduced risk of chronic GVHD but not with a reduced risk of relapse, NRM, or overall mortality. Routine testing of chimerism in the marrow and blood at 2 to 6 months after HCT with myeloablative conditioning regimens may be helpful in documenting engraftment in clinical trials, but provides only limited prognostic information in clinical practice.

  19. Early development of adipose cell lipogenesis and glycerol utilization in Zucker obese rats.

    PubMed

    Martin, R J; Lamprey, P M

    1975-05-01

    A study of adipose cell metabolism was made at ages 5, 7, 10, and 14 wk of age in genetically obese Zucker rats. Adipose samples were surgically removed and used for in vitro adipose cell incubations and for characterization of enzyme patterns. Lipogenic capacity from glucose and enzymes normally associated with lipogenesis (malic enzyme, citrate cleavage enzyme and glucose-6-PO4 dehydrogenase) followed the same pattern of development. At 5 wk of age, the adipose cells of obese animals had a greater capacity for fat synthesis than the lean rats. At all other ages lipogenic activity and enzyme levels were either similar or less than the pair-fed lean littermates. Glycerol utilization by isolated fat cells was similar; however, adipose tissue glycerokinase was elevated in obese rats at 14 wk of age. It was concluded that there was no apparent change in specific lipogenic capacity of fat cells from the obese rat when compared to its lean littermate. It was also concluded that increased adipose glycerokinase activity in obese rats represented a secondary shift in metabolism.

  20. Oncogenic Herpesvirus Utilizes Stress-Induced Cell Cycle Checkpoints for Efficient Lytic Replication.

    PubMed

    Balistreri, Giuseppe; Viiliäinen, Johanna; Turunen, Mikko; Diaz, Raquel; Lyly, Lauri; Pekkonen, Pirita; Rantala, Juha; Ojala, Krista; Sarek, Grzegorz; Teesalu, Mari; Denisova, Oxana; Peltonen, Karita; Julkunen, Ilkka; Varjosalo, Markku; Kainov, Denis; Kallioniemi, Olli; Laiho, Marikki; Taipale, Jussi; Hautaniemi, Sampsa; Ojala, Päivi M

    2016-02-01

    Kaposi's sarcoma herpesvirus (KSHV) causes Kaposi's sarcoma and certain lymphoproliferative malignancies. Latent infection is established in the majority of tumor cells, whereas lytic replication is reactivated in a small fraction of cells, which is important for both virus spread and disease progression. A siRNA screen for novel regulators of KSHV reactivation identified the E3 ubiquitin ligase MDM2 as a negative regulator of viral reactivation. Depletion of MDM2, a repressor of p53, favored efficient activation of the viral lytic transcription program and viral reactivation. During lytic replication cells activated a p53 response, accumulated DNA damage and arrested at G2-phase. Depletion of p21, a p53 target gene, restored cell cycle progression and thereby impaired the virus reactivation cascade delaying the onset of virus replication induced cytopathic effect. Herpesviruses are known to reactivate in response to different kinds of stress, and our study now highlights the molecular events in the stressed host cell that KSHV has evolved to utilize to ensure efficient viral lytic replication. PMID:26891221

  1. Oncogenic Herpesvirus Utilizes Stress-Induced Cell Cycle Checkpoints for Efficient Lytic Replication

    PubMed Central

    Turunen, Mikko; Diaz, Raquel; Lyly, Lauri; Pekkonen, Pirita; Rantala, Juha; Ojala, Krista; Sarek, Grzegorz; Teesalu, Mari; Denisova, Oxana; Peltonen, Karita; Julkunen, Ilkka; Varjosalo, Markku; Kainov, Denis; Kallioniemi, Olli; Laiho, Marikki; Taipale, Jussi; Hautaniemi, Sampsa; Ojala, Päivi M.

    2016-01-01

    Kaposi’s sarcoma herpesvirus (KSHV) causes Kaposi’s sarcoma and certain lymphoproliferative malignancies. Latent infection is established in the majority of tumor cells, whereas lytic replication is reactivated in a small fraction of cells, which is important for both virus spread and disease progression. A siRNA screen for novel regulators of KSHV reactivation identified the E3 ubiquitin ligase MDM2 as a negative regulator of viral reactivation. Depletion of MDM2, a repressor of p53, favored efficient activation of the viral lytic transcription program and viral reactivation. During lytic replication cells activated a p53 response, accumulated DNA damage and arrested at G2-phase. Depletion of p21, a p53 target gene, restored cell cycle progression and thereby impaired the virus reactivation cascade delaying the onset of virus replication induced cytopathic effect. Herpesviruses are known to reactivate in response to different kinds of stress, and our study now highlights the molecular events in the stressed host cell that KSHV has evolved to utilize to ensure efficient viral lytic replication. PMID:26891221

  2. Spontaneous calcification process in primary renal cells from a medullary sponge kidney patient harbouring a GDNF mutation.

    PubMed

    Mezzabotta, Federica; Cristofaro, Rosalba; Ceol, Monica; Del Prete, Dorella; Priante, Giovanna; Familiari, Alessandra; Fabris, Antonia; D'Angelo, Angela; Gambaro, Giovanni; Anglani, Franca

    2015-04-01

    Medullary nephrocalcinosis is a hallmark of medullary sponge kidney (MSK). We had the opportunity to study a spontaneous calcification process in vitro by utilizing the renal cells of a patient with MSK who was heterozygous for the c.-27 + 18G>A variant in the GDNF gene encoding glial cell-derived neurotrophic factor. The cells were obtained by collagenase digestion of papillary tissues from the MSK patient and from two patients who had no MSK or nephrocalcinosis. These cells were typed by immunocytochemistry, and the presence of mineral deposits was studied using von Kossa staining, scanning electron microscopy analysis and an ALP assay. Osteoblastic lineage markers were studied using immunocytochemistry and RT-PCR. Staminality markers were also analysed using flow cytometry, magnetic cell separation technology, immunocytochemistry and RT-PCR. Starting from p2, MSK and control cells formed nodules with a behaviour similar to that of calcifying pericytes; however, Ca₂PO₄ was only found in the MSK cultures. The MSK cells had morphologies and immunophenotypes resembling those of pericytes or stromal stem cells and were positive for vimentin, ZO1, αSMA and CD146. In addition, the MSK cells expressed osteocalcin and osteonectin, indicating an osteoblast-like phenotype. In contrast to the control cells, GDNF was down-regulated in the MSK cells. Stable GDNF knockdown was established in the HK2 cell line and was found to promote Ca₂PO₄ deposition when the cells were incubated with calcifying medium by regulating the osteonectin/osteopontin ratio in favour of osteonectin. Our data indicate that the human papilla may be a perivascular niche in which pericyte/stromal-like cells can undergo osteogenic differentiation under particular conditions and suggest that GDNF down-regulation may have influenced the observed phenomenon.

  3. Spontaneous calcification process in primary renal cells from a medullary sponge kidney patient harbouring a GDNF mutation

    PubMed Central

    Mezzabotta, Federica; Cristofaro, Rosalba; Ceol, Monica; Del Prete, Dorella; Priante, Giovanna; Familiari, Alessandra; Fabris, Antonia; D'Angelo, Angela; Gambaro, Giovanni; Anglani, Franca

    2015-01-01

    Medullary nephrocalcinosis is a hallmark of medullary sponge kidney (MSK). We had the opportunity to study a spontaneous calcification process in vitro by utilizing the renal cells of a patient with MSK who was heterozygous for the c.-27 + 18G>A variant in the GDNF gene encoding glial cell-derived neurotrophic factor. The cells were obtained by collagenase digestion of papillary tissues from the MSK patient and from two patients who had no MSK or nephrocalcinosis. These cells were typed by immunocytochemistry, and the presence of mineral deposits was studied using von Kossa staining, scanning electron microscopy analysis and an ALP assay. Osteoblastic lineage markers were studied using immunocytochemistry and RT-PCR. Staminality markers were also analysed using flow cytometry, magnetic cell separation technology, immunocytochemistry and RT-PCR. Starting from p2, MSK and control cells formed nodules with a behaviour similar to that of calcifying pericytes; however, Ca2PO4 was only found in the MSK cultures. The MSK cells had morphologies and immunophenotypes resembling those of pericytes or stromal stem cells and were positive for vimentin, ZO1, αSMA and CD146. In addition, the MSK cells expressed osteocalcin and osteonectin, indicating an osteoblast-like phenotype. In contrast to the control cells, GDNF was down-regulated in the MSK cells. Stable GDNF knockdown was established in the HK2 cell line and was found to promote Ca2PO4 deposition when the cells were incubated with calcifying medium by regulating the osteonectin/osteopontin ratio in favour of osteonectin. Our data indicate that the human papilla may be a perivascular niche in which pericyte/stromal-like cells can undergo osteogenic differentiation under particular conditions and suggest that GDNF down-regulation may have influenced the observed phenomenon. PMID:25692823

  4. The Establishment of a Production-ready Manufacturing Process Utilizing Thin Silicon Substrates for Solar Cells

    NASA Technical Reports Server (NTRS)

    Pryor, R. A.

    1979-01-01

    During the months of February and March, work towards the goals of the contract were started as scheduled. The first shipment of thin substrates were received and wafer processing was initiated. The objective of the contract is to investigate, develop and characterize the methods for establishing a production-ready manufacturing process which utilizes thin silicon substrates for solar cells. The thin substrates to be manufactured are three inches diameter, p-type Czochralski wafers of approximately 1 Omega cm resistivity. The wafers are prepared by sawing directly to thickness of 8 mils and 5 mils. To ensure removal of residual saw damage, most substrates are chemically etched to final thicknesses of 7 mils and 4 mils. The thin substrates are used to fabricate solar cells by standard processing techniques.

  5. Electrodes and electrochemical storage cells utilizing tin-modified active materials

    DOEpatents

    Anani, Anaba; Johnson, John; Lim, Hong S.; Reilly, James; Schwarz, Ricardo; Srinivasan, Supramaniam

    1995-01-01

    An electrode has a substrate and a finely divided active material on the substrate. The active material is ANi.sub.x-y-z Co.sub.y Sn.sub.z, wherein A is a mischmetal or La.sub.1-w M.sub.w, M is Ce, Nd, or Zr, w is from about 0.05 to about 1.0, x is from about 4.5 to about 5.5, y is from 0 to about 3.0, and z is from about 0.05 to about 0.5. An electrochemical storage cell utilizes such an electrode as the anode. The storage cell further has a cathode, a separator between the cathode and the anode, and an electrolyte.

  6. Single cell kinetics of phenotypic switching in the arabinose utilization system of E. coli.

    PubMed

    Fritz, Georg; Megerle, Judith A; Westermayer, Sonja A; Brick, Delia; Heermann, Ralf; Jung, Kirsten; Rädler, Joachim O; Gerland, Ulrich

    2014-01-01

    Inducible switching between phenotypes is a common strategy of bacteria to adapt to fluctuating environments. Here, we analyze the switching kinetics of a paradigmatic inducible system, the arabinose utilization system in E. coli. Using time-lapse fluorescence microscopy of microcolonies in a microfluidic chamber, which permits sudden up- and down-shifts in the inducer arabinose, we characterize the single-cell gene expression dynamics of the araBAD operon responsible for arabinose degradation. While there is significant, inducer-dependent cell-to-cell variation in the timing of the on-switching, the off-switching triggered by sudden removal of arabinose is homogeneous and rapid. We find that rapid off-switching does not depend on internal arabinose degradation. Because the system is regulated via the internal arabinose level sensed by AraC, internal arabinose must be rapidly depleted by leakage or export from the cell, or by degradation via a non-canonical pathway. We explored whether the poorly characterized membrane protein AraJ, which is part of the arabinose regulon and has been annotated as a possible arabinose efflux protein, is responsible for rapid depletion. However, we find that AraJ is not essential for rapid switching to the off-state. We develop a mathematical model for the arabinose system, which quantitatively describes both the heterogeneous on-switching and the homogeneous off-switching. The model also predicts that mutations which disrupt the positive feedback of internal arabinose on the production of arabinose uptake proteins change the heterogeneous on-switching behavior into a homogeneous, graded response. We construct such a mutant and confirm the graded response experimentally. Taken together, our results indicate that the physiological switching behavior of this sugar utilization system is asymmetric, such that off-switching is always rapid and homogeneous, while on-switching is slow and heterogeneously timed at sub-saturating inducer

  7. Hsp90 Maintains Proteostasis of the Galactose Utilization Pathway To Prevent Cell Lethality

    PubMed Central

    Gopinath, Rajaneesh Karimpurath

    2016-01-01

    Hsp90 is a molecular chaperone that aids in the folding of its metastable client proteins. Past studies have shown that it can exert a strong impact on some cellular pathways by controlling key regulators. However, it is unknown whether several components of a single pathway are collectively regulated by Hsp90. Here, we observe that Hsp90 influences the protein abundance of multiple Gal proteins and the efficiency of galactose utilization even after the galactose utilization pathway (GAL pathway) is fully induced. The effect of Hsp90 on Gal proteins is not at the transcriptional level. Moreover, Gal1 is found to physically interact with Hsp90, and its stability is reduced in low-Hsp90 cells. When Hsp90 is compromised, several Gal proteins form protein aggregates that colocalize with the disaggregase Hsp104. These results suggest that Gal1 and other Gal proteins are probably the clients of Hsp90. An unbalanced GAL pathway has been known to cause fatal growth arrest due to accumulation of toxic galactose metabolic intermediates. It is likely that Hsp90 chaperones multiple Gal proteins to maintain proteostasis and prevent cell lethality especially in a fluctuating environment. PMID:26951197

  8. Making the grid the backup: Utility applications for fuel cell power

    SciTech Connect

    Eklof, S.L.

    1996-12-31

    Fuel cells are recognized as a versatile power generation option and accepted component of SMUD`s ART Program. SMUD has received wide support and recognition for promoting and implementing fuel cell power plants, as well as other innovative generation, based primarily on technological factors. Current economic and technical realities in the electric generation market highlight other important factors, such as the cost involved to develop a slate of such resources. The goal now is to develop only those select quality resources most likely to become commercially viable in the near future. The challenge becomes the identification of candidate technologies with the greatest potential, and then matching the technologies with the applications that will help to make them successful. Utility participation in this development is critical so as to provide the industry with case examples of advanced technologies that can be applied in a way beneficial to both the utility and its customers. The ART resource acquisitions provide the experience base upon which to guide this selection process, and should bring about the cost reductions and reliability improvements sought.

  9. Developing RCM Strategy for Hydrogen Fuel Cells Utilizing On Line E-Condition Monitoring

    NASA Astrophysics Data System (ADS)

    Baglee, D.; Knowles, M. J.

    2012-05-01

    Fuel cell vehicles are considered to be a viable solution to problems such as carbon emissions and fuel shortages for road transport. Proton Exchange Membrane (PEM) Fuel Cells are mainly used in this purpose because they can run at low temperatures and have a simple structure. Yet high maintenance costs and the inherent dangers of maintaining equipment using hydrogen are two main issues which need to be addressed. The development of appropriate and efficient strategies is currently lacking with regard to fuel cell maintenance. A Reliability Centered Maintenance (RCM) approach offers considerable benefit to the management of fuel cell maintenance since it includes an identification and consideration of the impact of critical components. Technological developments in e-maintenance systems, radio-frequency identification (RFID) and personal digital assistants (PDAs) have proven to satisfy the increasing demand for improved reliability, efficiency and safety. RFID technology is used to store and remotely retrieve electronic maintenance data in order to provide instant access to up-to-date, accurate and detailed information. The aim is to support fuel cell maintenance decisions by developing and applying a blend of leading-edge communications and sensor technology including RFID. The purpose of this paper is to review and present the state of the art in fuel cell condition monitoring and maintenance utilizing RCM and RFID technologies. Using an RCM analysis critical components and fault modes are identified. RFID tags are used to store the critical information, possible faults and their cause and effect. The relationship between causes, faults, symptoms and long term implications of fault conditions are summarized. Finally conclusions are drawn regarding suggested maintenance strategies and the optimal structure for an integrated, cost effective condition monitoring and maintenance management system.

  10. Quantification of cell viability and rapid screening anti-cancer drug utilizing nanomechanical fluctuation.

    PubMed

    Wu, Shangquan; Liu, Xiaoli; Zhou, Xiarong; Liang, Xin M; Gao, Dayong; Liu, Hong; Zhao, Gang; Zhang, Qingchuan; Wu, Xiaoping

    2016-03-15

    Cancer is a serious threat to human health. Although numerous anti-cancer drugs are available clinically, many have shown toxic side effects due to poor tumor-selectivity, and reduced effectiveness due to cancers rapid development of resistance to treatment. The development of new highly efficient and practical methods to quantify cell viability and its change under drug treatment is thus of significant importance in both understanding of anti-cancer mechanism and anti-cancer drug screening. Here, we present an approach of utilizing a nanomechanical fluctuation based highly sensitive microcantilever sensor, which is capable of characterizing the viability of cells and quantitatively screening (within tens of minutes) their responses to a drug with the obvious advantages of a rapid, label-free, quantitative, noninvasive, real-time and in-situ assay. The microcantilever sensor operated in fluctuation mode was used in evaluating the paclitaxel effectiveness on breast cancer cell line MCF-7. This study demonstrated that the nanomechanical fluctuations of the microcantilever sensor are sensitive enough to detect the dynamic variation in cellular force which is provided by the cytoskeleton, using cell metabolism as its energy source, and the dynamic instability of microtubules plays an important role in the generation of the force. We propose that cell viability consists of two parts: biological viability and mechanical viability. Our experimental results suggest that paclitaxel has little effect on biological viability, but has a significant effect on mechanical viability. This new method provides a new concept and strategy for the evaluation of cell viability and the screening of anti-cancer drugs.

  11. Quantification of cell viability and rapid screening anti-cancer drug utilizing nanomechanical fluctuation.

    PubMed

    Wu, Shangquan; Liu, Xiaoli; Zhou, Xiarong; Liang, Xin M; Gao, Dayong; Liu, Hong; Zhao, Gang; Zhang, Qingchuan; Wu, Xiaoping

    2016-03-15

    Cancer is a serious threat to human health. Although numerous anti-cancer drugs are available clinically, many have shown toxic side effects due to poor tumor-selectivity, and reduced effectiveness due to cancers rapid development of resistance to treatment. The development of new highly efficient and practical methods to quantify cell viability and its change under drug treatment is thus of significant importance in both understanding of anti-cancer mechanism and anti-cancer drug screening. Here, we present an approach of utilizing a nanomechanical fluctuation based highly sensitive microcantilever sensor, which is capable of characterizing the viability of cells and quantitatively screening (within tens of minutes) their responses to a drug with the obvious advantages of a rapid, label-free, quantitative, noninvasive, real-time and in-situ assay. The microcantilever sensor operated in fluctuation mode was used in evaluating the paclitaxel effectiveness on breast cancer cell line MCF-7. This study demonstrated that the nanomechanical fluctuations of the microcantilever sensor are sensitive enough to detect the dynamic variation in cellular force which is provided by the cytoskeleton, using cell metabolism as its energy source, and the dynamic instability of microtubules plays an important role in the generation of the force. We propose that cell viability consists of two parts: biological viability and mechanical viability. Our experimental results suggest that paclitaxel has little effect on biological viability, but has a significant effect on mechanical viability. This new method provides a new concept and strategy for the evaluation of cell viability and the screening of anti-cancer drugs. PMID:26406457

  12. Ceruloplasmin copper induces oxidant damage by a redox process utilizing cell-derived superoxide as reductant

    NASA Technical Reports Server (NTRS)

    Mukhopadhyay, C. K.; Fox, P. L.

    1998-01-01

    Oxidative damage by transition metals bound to proteins may be an important pathogenic mechanism. Ceruloplasmin (Cp) is a Cu-containing plasma protein thought to be involved in oxidative modification of lipoproteins. We have previously shown that Cp increased cell-mediated low-density lipoprotein (LDL) oxidation by a process requiring cell-derived superoxide, but the underlying chemical mechanism(s) is (are) unknown. We now show that superoxide reduction of Cp Cu is a critical reaction in cellular LDL oxidation. By bathocuproine disulfonate (BCS) binding and by superoxide utilization, we showed that exogenous superoxide reduces a single Cp Cu atom, the same Cu required for LDL oxidation. The Cu atom remained bound to Cp during the redox cycle. Three avenues of evidence showed that vascular cells reduce Cp Cu by a superoxide-dependent process. The 2-fold higher rate of Cp Cu reduction by smooth muscle cells (SMC) compared to endothelial cells (EC) was consistent with their relative rates of superoxide release. Furthermore, Cp Cu reduction by cells was blocked by Cu,Zn superoxide dismutase (SOD1). Finally, the level of superoxide produced by EC and SMC was sufficient to cause the amount of Cu reduction observed. An important role of Cp Cu reduction in LDL oxidation was suggested by results showing that SOD1 inhibited Cp Cu reduction and LDL oxidation by SMC with equal potency, while tumor necrosis factor-alpha stimulated both processes. In summary, these results show that superoxide is a critical cellular reductant of divalent transition metals involved in oxidation, and that protein-bound Cu is a substrate for this reaction. The role of these mechanisms in oxidative processes in vivo has yet to be defined.

  13. Clinical utility of circulating tumor cell counting through CellSearch®: the dilemma of a concept suspended in Limbo

    PubMed Central

    Raimondi, Cristina; Gradilone, Angela; Naso, Giuseppe; Cortesi, Enrico; Gazzaniga, Paola

    2014-01-01

    To date, 10 years after the first demonstration of circulating tumor cells (CTCs), prognostic significance in metastatic breast cancer using the US Food and Drug Administration–cleared system CellSearch®, the potential utility of CTCs in early clinical development of drugs, their role as a surrogate marker of response to therapy, and their molecular analysis for patient stratification for targeted therapies are still major unsolved questions. Great expectations are pinned on the ongoing interventional trials aimed to demonstrate that CTCs might be of value for guiding treatment of patients and predicting cancer progression. To fill the gap between theory and practice with regard to the clinical utility of CTCs, a bridge is needed, taking into account innovative design for clinical trials, a revised definition of traditional CTCs, next-generation CTC technology, the potential clinical application of CTC analysis in non-validated settings of disease, and finally, expanding the number of patients enrolled in the studies. In this regard, the results of the first European pooled analysis definitely validated the independent prognostic value of CTC counting in metastatic breast cancer patients. PMID:24790460

  14. Optical Frequency Optimization of a High Intensity Laser Power Beaming System Utilizing VMJ Photovoltaic Cells

    NASA Technical Reports Server (NTRS)

    Raible, Daniel E.; Dinca, Dragos; Nayfeh, Taysir H.

    2012-01-01

    An effective form of wireless power transmission (WPT) has been developed to enable extended mission durations, increased coverage and added capabilities for both space and terrestrial applications that may benefit from optically delivered electrical energy. The high intensity laser power beaming (HILPB) system enables long range optical 'refueling" of electric platforms such as micro unmanned aerial vehicles (MUAV), airships, robotic exploration missions and spacecraft platforms. To further advance the HILPB technology, the focus of this investigation is to determine the optimal laser wavelength to be used with the HILPB receiver, which utilizes vertical multi-junction (VMJ) photovoltaic cells. Frequency optimization of the laser system is necessary in order to maximize the conversion efficiency at continuous high intensities, and thus increase the delivered power density of the HILPB system. Initial spectral characterizations of the device performed at the NASA Glenn Research Center (GRC) indicate the approximate range of peak optical-to-electrical conversion efficiencies, but these data sets represent transient conditions under lower levels of illumination. Extending these results to high levels of steady state illumination, with attention given to the compatibility of available commercial off-the-shelf semiconductor laser sources and atmospheric transmission constraints is the primary focus of this paper. Experimental hardware results utilizing high power continuous wave (CW) semiconductor lasers at four different operational frequencies near the indicated band gap of the photovoltaic VMJ cells are presented and discussed. In addition, the highest receiver power density achieved to date is demonstrated using a single photovoltaic VMJ cell, which provided an exceptionally high electrical output of 13.6 W/sq cm at an optical-to-electrical conversion efficiency of 24 percent. These results are very promising and scalable, as a potential 1.0 sq m HILPB receiver of

  15. Utility of Lymphoblastoid Cell Lines for Induced Pluripotent Stem Cell Generation

    PubMed Central

    Kumar, Satish; Curran, Joanne E.; Glahn, David C.; Blangero, John

    2016-01-01

    A large number of EBV immortalized LCLs have been generated and maintained in genetic/epidemiological studies as a perpetual source of DNA and as a surrogate in vitro cell model. Recent successes in reprograming LCLs into iPSCs have paved the way for generating more relevant in vitro disease models using this existing bioresource. However, the overall reprogramming efficiency and success rate remain poor and very little is known about the mechanistic changes that take place at the transcriptome and cellular functional level during LCL-to-iPSC reprogramming. Here, we report a new optimized LCL-to-iPSC reprogramming protocol using episomal plasmids encoding pluripotency transcription factors and mouse p53DD (p53 carboxy-terminal dominant-negative fragment) and commercially available reprogramming media. We achieved a consistently high reprogramming efficiency and 100% success rate using this optimized protocol. Further, we investigated the transcriptional changes in mRNA and miRNA levels, using FC-abs ≥ 2.0 and FDR ≤ 0.05 cutoffs; 5,228 mRNAs and 77 miRNAs were differentially expressed during LCL-to-iPSC reprogramming. The functional enrichment analysis of the upregulated genes and activation of human pluripotency pathways in the reprogrammed iPSCs showed that the generated iPSCs possess transcriptional and functional profiles very similar to those of human ESCs. PMID:27375745

  16. p16INK4a immunocytochemistry versus human papillomavirus testing for triage of women with minor cytologic abnormalities: a systematic review and meta-analysis.

    PubMed

    Roelens, Jolien; Reuschenbach, Miriam; von Knebel Doeberitz, Magnus; Wentzensen, Nicolas; Bergeron, Christine; Arbyn, Marc

    2012-10-25

    The best method for identifying women who have minor cervical lesions that require diagnostic workup remains unclear. The authors of this report performed a meta-analysis to assess the accuracy of cyclin-dependent kinase inhibitor 2A (p16(INK4a)) immunocytochemistry compared with high-risk human papillomavirus DNA testing with Hybrid Capture 2 (HC2) to detect grade 2 or greater cervical intraepithelial neoplasia (CIN2+) and CIN3+ among women who had cervical cytology indicating atypical squamous cells of undetermined significance (ASC-US) or low-grade cervical lesions (LSIL). A literature search was performed in 3 electronic databases to identify studies that were eligible for this meta-analysis. Seventeen studies were included in the meta-analysis. The pooled sensitivity of p16(INK4a) to detect CIN2+ was 83.2% (95% confidence interval [CI], 76.8%-88.2%) and 83.8% (95% CI, 73.5%-90.6%) in ASC-US and LSIL cervical cytology, respectively, and the pooled specificities were 71% (95% CI, 65%-76.4%) and 65.7% (95% CI, 54.2%-75.6%), respectively. Eight studies provided both HC2 and p16(INK4a) triage data. p16(INK4a) and HC2 had similar sensitivity, and p16(INK4a) has significantly higher specificity in the triage of women with ASC-US (relative sensitivity, 0.95 [95% CI, 0.89-1.01]; relative specificity, 1.82 [95% CI, 1.57-2.12]). In the triage of LSIL, p16(INK4a) had significantly lower sensitivity but higher specificity compared with HC2 (relative sensitivity, 0.87 [95% CI, 0.81-0.94]; relative specificity, 2.74 [95% CI, 1.99-3.76]). The published literature indicated the improved accuracy of p16(INK4a) compared with HC2 testing in the triage of women with ASC-US. In LSIL triage, p16(INK4a) was more specific but less sensitive.

  17. p16INK4a immunocytochemistry versus HPV testing for triage of women with minor cytological abnormalities: A systematic review and meta-analysis

    PubMed Central

    Roelens, Jolien; Reuschenbach, Miriam; von Knebel-Doeberitz, Magnus; Wentzensen, Nicolas; Bergeron, Christine; Arbyn, Marc

    2014-01-01

    Background The best method to identify women with minor cervical lesions that require diagnostic work-up remains unclear. We performed a meta-analysis to assess the accuracy of p16INK4a immunocytochemistry compared to hrHPV DNA testing with hybrid capture II (HC2) to detect cervical intraepithelial neoplasia (CIN2+ and CIN3+) in women with a cervical cytology showing atypical squamous cells of undetermined significance (ASC-US) or low-grade cervical lesions (LSIL). Methods A literature search was performed in three electronic databases to identify studies eligible for this meta-analysis. Results Seventeen studies were included in the meta-analysis. The pooled sensitivity of p16INK4a to detect CIN2+ was 83.2% (95%CI: 76.8–88.2%) and 83.8% (95%CI: 73.5–90.6%) in ASC-US and LSIL cervical cytology respectively; pooled specificities were 71.0% (95%CI: 65.0–76.4%) and 65.7% (95%CI: 54.2–75.6%). Eight studies provided both HC2 and p16INK4a triage data. p16INK4a and HC2 have a similar sensitivity and p16INK4a has significantly higher specificity in the triage of women with ASC-US (relative sensitivity: 0.95 (95%CI: 0.89–1.01); relative specificity: 1.82 (95%CI: 1.57–2.12)). In the triage of LSIL, p16INK4a has a significantly lower sensitivity but higher specificity compared to HC2 (relative sensitivity: 0.87 (95%CI: 0.81–0.94); relative specificity: 2.74 (1.99–3.76)). Conclusion The published literature indicates an improved accuracy of p16INK4a compared to HC2 testing in the triage of ASC-US. In LSIL triage p16INK4a is more specific but less sensitive. PMID:22700382

  18. Hydrogen Storage in Diamond Powder Utilizing Plasma NaF Surface Treatment for Fuel Cell Applications

    SciTech Connect

    Leal, David A.; Leal-Quiros, E.; Velez, Angel; Prelas, Mark A.; Gosh, Tushar

    2006-12-04

    Hydrogen Fuel Cells offer the vital solution to the world's socio-political dependence on oil. Due to existing difficulty in safe and efficient hydrogen storage for fuel cells, storing the hydrogen in hydrocarbon compounds such as artificial diamond is a realistic solution. By treating the surface of the diamond powder with a Sodium Fluoride plasma exposure, the surface of the diamond is cleaned of unwanted molecules. Due to fluorine's electro negativity, the diamond powder is activated and ready for hydrogen absorption. These diamond powder pellets are then placed on a graphite platform that is heated by conduction in a high voltage circuit made of tungsten wire. Then, the injection of hydrogen gas into chamber allows the storage of the Hydrogen on the surface of the diamond powder. By neutron bombardment in the nuclear reactor, or Prompt Gamma Neutron Activation Analysis, the samples are examined for parts per million amounts of hydrogen in the sample. Sodium Fluoride surface treatment allows for higher mass percentage of stored hydrogen in a reliable, resistant structure, such as diamond for fuel cells and permanently alters the diamonds terminal bonds for re-use in the effective storage of hydrogen. The highest stored amount utilizing the NaF plasma surface treatment was 22229 parts per million of hydrogen in the diamond powder which amounts to 2.2229% mass increase.

  19. Harvest and utilization of chemical energy in wastes by microbial fuel cells.

    PubMed

    Sun, Min; Zhai, Lin-Feng; Li, Wen-Wei; Yu, Han-Qing

    2016-05-21

    Organic wastes are now increasingly viewed as a resource of energy that can be harvested by suitable biotechnologies. One promising technology is microbial fuel cells (MFC), which can generate electricity from the degradation of organic pollutants. While the environmental benefits of MFC in waste treatment have been recognized, their potential as an energy producer is not fully understood. Although progresses in material and engineering have greatly improved the power output from MFC, how to efficiently utilize the MFC's energy in real-world scenario remains a challenge. In this review, fundamental understandings on the energy-generating capacity of MFC from real waste treatment are provided and the challenges and opportunities are discussed. The limiting factors restricting the energy output and impairing the long-term reliability of MFC are also analyzed. Several energy storage and in situ utilization strategies for the management of MFC's energy are proposed, and future research needs for real-world application of this approach are explored. PMID:26936021

  20. Assessment of soft tissue hemangiomas in children utilizing Tc-99m labelled red blood cells

    SciTech Connect

    Miller, J.H.

    1984-01-01

    Hemangiomas may present in infancy as soft tissue masses. Occasionally these lesions may be extensive or may not be clinically recognized as a hemangioma, often causing concern for the presence of a malignant lesion. In later childhood these lesions, which may be occult, may cause overgrowth of an extremity. Evaluation of soft tissue masses suspected of being a hemangioma utilizing Technetium 99m labelled red blood cells has been very valuable. This method allows a dynamic evaluation of first pass blood flow. Subsequent static scintiphotos allow an assessment of the lesion itself. These scintiphotos may be obtained sequentially to evaluate therapy. Twenty patients were evaluated by this method ranging in age from two months to eleven years. There were 13 females and seven males. Lesions evaluated by this method include six hemangiomas of the head and neck: parotic region (2), facial (3), and tongue (1). Extremity lesions were evaluated in six children including both upper extremity (1) and lower extremity (5). Torso lesions evaluated include chest wall (2), abdominal wall (2), and one hemangioma of the gut. This procedure is quickly performed on an outpatient basis, has high anatomic resolution, provides and assessment of these lesions in a manner not available by any other imaging procedure and usually requires no sedation. The radiation exposure for this procedure is low (approximately, a 400mR total body dose) and has been well tolerated by both patients and their parents. Scintigraphic evaluation should be the first diagnostic method utilized in the evaluation of these lesions.

  1. Do Circulating Tumor Cells, Exosomes, and Circulating Tumor Nucleic Acids Have Clinical Utility?

    PubMed Central

    Gold, Bert; Cankovic, Milena; Furtado, Larissa V.; Meier, Frederick; Gocke, Christopher D.

    2016-01-01

    Diagnosing and screening for tumors through noninvasive means represent an important paradigm shift in precision medicine. In contrast to tissue biopsy, detection of circulating tumor cells (CTCs) and circulating tumor nucleic acids provides a minimally invasive method for predictive and prognostic marker detection. This allows early and serial assessment of metastatic disease, including follow-up during remission, characterization of treatment effects, and clonal evolution. Isolation and characterization of CTCs and circulating tumor DNA (ctDNA) are likely to improve cancer diagnosis, treatment, and minimal residual disease monitoring. However, more trials are required to validate the clinical utility of precise molecular markers for a variety of tumor types. This review focuses on the clinical utility of CTCs and ctDNA testing in patients with solid tumors, including somatic and epigenetic alterations that can be detected. A comparison of methods used to isolate and detect CTCs and some of the intricacies of the characterization of the ctDNA are also provided. PMID:25908243

  2. Using circulating tumor cells to inform on prostate cancer biology and clinical utility

    PubMed Central

    Li, Jing; Gregory, Simon G.; Garcia-Blanco, Mariano A.; Armstrong, Andrew J.

    2016-01-01

    Substantial advances in the molecular biology of prostate cancer have led to the approval of multiple new systemic agents to treat men with metastatic castration-resistant prostate cancer (mCRPC). These treatments encompass androgen receptor directed therapies, immunotherapies, bone targeting radiopharmaceuticals and cytotoxic chemotherapies. There is, however, great heterogeneity in the degree of patient benefit with these agents, thus fueling the need to develop predictive biomarkers that are able to rationally guide therapy. Circulating tumor cells (CTCs) have the potential to provide an assessment of tumor-specific biomarkers through a non-invasive, repeatable “liquid biopsy” of a patient’s cancer at a given point in time. CTCs have been extensively studied in men with mCRPC, where CTC enumeration using the Cellsearch® method has been validated and FDA approved to be used in conjunction with other clinical parameters as a prognostic biomarker in metastatic prostate cancer. In addition to enumeration, more sophisticated molecular profiling of CTCs is now feasible and may provide more clinical utility as it may reflect tumor evolution within an individual particularly under the pressure of systemic therapies. Here, we review technologies used to detect and characterize CTCs, and the potential biological and clinical utility of CTC molecular profiling in men with metastatic prostate cancer. PMID:26079252

  3. Fabrication of polymer electrolyte membrane fuel cell MEAs utilizing inkjet print technology

    NASA Astrophysics Data System (ADS)

    Towne, Silas; Viswanathan, Vish; Holbery, James; Rieke, Peter

    Utilizing drop-on-demand technology, we have successfully fabricated hydrogen-air polymer electrolyte membrane fuel cells (PEMFC), demonstrated some of the processing advantages of this technology and have demonstrated that the performance is comparable to conventionally fabricated membrane electrode assemblies (MEAs). Commercial desktop inkjet printers were used to deposit the active catalyst electrode layer directly from print cartridges onto Nafion ® polymer membranes in the hydrogen form. The layers were well-adhered and withstood simple tape peel, bending and abrasion tests and did so without any post-deposition hot press step. The elimination of this processing step suggests that inkjet-based fabrication or similar processing technologies may provide a route to less expensive large-scale fabrication of PEMFCs. When tested in our experimental apparatus, open circuit voltages up to 0.87 V and power densities of up to 155 mW cm -2 were obtained with a catalyst loading of 0.20 mg Pt cm -2. A commercially available membrane under identical, albeit not optimized test conditions, showed about 7% greater power density. The objective of this work was to demonstrate some of the processing advantages of drop-on-demand technology for fabrication of MEAs. It remains to be determined if inkjet fabrication offers performance advantages or leads to more efficient utilization of expensive catalyst materials.

  4. Timing and Dynamics of Single Cell Gene Expression in the Arabinose Utilization System

    NASA Astrophysics Data System (ADS)

    Megerle, J.; Fritz, G.; Gerland, U.; Jung, K.; Radler, J.

    2008-08-01

    The arabinose utilization system of E. coli displays a stochastic "all or nothing" response at intermediate levels of arabinose, where the population divides into a fraction catabolizing the sugar at a high rate (ON state) and a fraction not utilizing arabinose (OFF state). Here we study this decision process in individual cells, focusing on the dynamics of the transition from the OFF to the ON state. Using quantitative time-lapse microscopy, we determine the time delay between inducer addition and fluorescence onset of a GFP reporter. Through independent characterization of the GFP maturation process, we can separate the lag time caused by the reporter from the intrinsic activation time of the arabinose system. The resulting distribution of intrinsic time delays scales inversely with the external arabinose concentration, and is compatible with a simple stochastic model for arabinose uptake. Our findings support the idea that the heterogeneous timing of gene induction is causally related to a broad distribution of uptake proteins at the time of sugar addition.

  5. Copper catalysis for enhancement of cobalt leaching and acid utilization efficiency in microbial fuel cells.

    PubMed

    Liu, Yaxuan; Shen, Jingya; Huang, Liping; Wu, Dan

    2013-11-15

    Enhancement of both cobalt leaching from LiCoO2 and acid utilization efficiency (AUE) in microbial fuel cells (MFCs) was successfully achieved by the addition of Cu(II). A dosage of 10mg/L Cu(II) improved both cobalt leaching up to 308% and AUE of 171% compared to the controls with no presence of Cu(II). The apparent activation energy of cobalt leaching catalyzed by Cu(II) in MFCs was only 11.8 kJ/mol. These results demonstrate cobalt leaching in MFCs using Cu(II) as a catalyst may be an effective strategy for cobalt recovery and recycle of spent Li-ion batteries, and the evidence of influence factors including solid/liquid ratio, temperature, and pH and solution conductivity can contribute to improving understanding of and optimizing cobalt leaching catalyzed by Cu(II) in MFCs.

  6. Copper catalysis for enhancement of cobalt leaching and acid utilization efficiency in microbial fuel cells.

    PubMed

    Liu, Yaxuan; Shen, Jingya; Huang, Liping; Wu, Dan

    2013-11-15

    Enhancement of both cobalt leaching from LiCoO2 and acid utilization efficiency (AUE) in microbial fuel cells (MFCs) was successfully achieved by the addition of Cu(II). A dosage of 10mg/L Cu(II) improved both cobalt leaching up to 308% and AUE of 171% compared to the controls with no presence of Cu(II). The apparent activation energy of cobalt leaching catalyzed by Cu(II) in MFCs was only 11.8 kJ/mol. These results demonstrate cobalt leaching in MFCs using Cu(II) as a catalyst may be an effective strategy for cobalt recovery and recycle of spent Li-ion batteries, and the evidence of influence factors including solid/liquid ratio, temperature, and pH and solution conductivity can contribute to improving understanding of and optimizing cobalt leaching catalyzed by Cu(II) in MFCs. PMID:24007993

  7. Utilization of hydrolysate from lignocellulosic biomass pretreatment to generate electricity by enzymatic fuel cell system.

    PubMed

    Kim, Sung Bong; Kim, Dong Sup; Yang, Ji Hyun; Lee, Junyoung; Kim, Seung Wook

    2016-04-01

    The waste hydrolysate after dilute acid pretreatment (DAP) of lignocellulosic biomass was utilized to generate electricity using an enzymatic fuel cell (EFC) system. During DAP, the components of biomass containing hemicellulose and other compounds are hydrolyzed, and glucose is solubilized into the dilute acid solution, called as the hydrolysate liquid. Glucose oxidase (GOD) and laccase (Lac) were assembled on the electrode of the anode and cathode, respectively. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were measured, and the maximum power density was found to be 1.254×10(3) μW/cm(2). The results indicate that the hydrolysate from DAP is a reliable electrolyte containing the fuel of EFC. Moreover, the impurities in the hydrolysate such as phenols and furans slightly affected the charge transfer on the surface of the electrode, but did not affect the power generation of the EFC system in principal. PMID:26920478

  8. Engineering aptazyme switches for conditional gene expression in mammalian cells utilizing an in vivo screening approach.

    PubMed

    Rehm, Charlotte; Klauser, Benedikt; Hartig, Jörg S

    2015-01-01

    Artificial RNA switches are an emerging class of genetic controllers suitable for synthetic biology applications. Aptazymes are fusions composed of an aptamer domain and a self-cleaving ribozyme. The utilization of aptazymes for conditional gene expression displays several advantages over employing conventional transcription factor-based techniques as aptazymes require minimal genomic space, fulfill their function without the need of protein cofactors, and most importantly are reprogrammable with respect to ligand selectivity and the RNA function to be regulated. Technologies that enable the generation of aptazymes to defined input ligands are of interest for the construction of biocomputing devices and biosensing applications. In this chapter we present a method that facilitates the in vivo screening of randomized pools of aptazymes in mammalian cells. PMID:25967058

  9. Production of cytidine 5'-diphosphorylcholine with high utilization of ATP by whole cells of Saccharomyces cerevisiae.

    PubMed

    Tang, Jiapeng; Chen, Yong; Chen, Xiaochun; Yao, Yuelan; Ying, Hanjie; Xiong, Jian; Bai, Jianxin

    2010-11-01

    Cytidine 5'-diphosphorylcholine (CDP-choline) was produced using a high efficiency ATP regeneration system and the Kennedy pathway in whole cells of Saccharomyces cerevisiae As 2.398. Out of eight variables, KH(2)PO(4), glycerol and (NH(4))(2)SO(4) were considered to be the most significant factors by response surface methodology including a Plackett-Burman design, path of steepest accent and central composite design. The optimum levels of the three variables were 20.13g/L KH(2)PO(4), 12.35g/L glycerol and 0.49g/L (NH(4))(2)SO(4), respectively. Energy utilization efficiency increased from 10.59% to 16.72% and choline chloride conversion yields increased from 12.35% to 42.78%. A high efficiency ATP regeneration system improves CDP-choline production.

  10. SPOUTED BED ELECTRODES (SBE) FOR DIRECT UTILIZATION OF CARBON IN FUEL CELLS

    SciTech Connect

    J.M. Calo

    2004-12-01

    This Phase I project was focused on an investigation of spouted bed particulate electrodes for the direct utilization of solid carbon in fuel cells. This approach involves the use of a circulating carbon particle/molten carbonate slurry in the cell that provides a few critical functions: it (1) fuels the cell continuously with entrained carbon particles; (2) brings particles to the anode surfaces hydrodynamically; (3) removes ash from the anode surfaces and the cell hydrodynamically; (4) provides a facile means of cell temperature control due to its large thermal capacitance; (5) provides for electrolyte maintenance and control in the electrode separator(s); and (6) can (potentially) improve carbon conversion rates by ''pre-activating'' carbon particle surfaces via formation of intermediate oxygen surface complexes in the bulk molten carbonate. The approach of this scoping project was twofold: (1) adaptation and application of a CFD code, originally developed to simulate particle circulation in spouted bed electrolytic reactors, to carbon particle circulation in DCFC systems; and (2) experimental investigation of the hydrodynamics of carbon slurry circulation in DCFC systems using simulated slurry mixtures. The CFD model results demonstrated that slurry recirculation can be used to hydrodynamically feed carbon particles to anode surfaces. Variations of internal configurations were investigated in order to explore effects on contacting. It was shown that good contacting with inclined surfaces could be achieved even when the particles are of the same density as the molten carbonate. The use of CO{sub 2} product gas from the fuel cell as a ''lift-gas'' to circulate the slurry was also investigated with the model. The results showed that this is an effective method of slurry circulation; it entrains carbon particles more effectively in the draft duct and produces a somewhat slower recirculation rate, and thus higher residence times on anode surfaces, and can be

  11. Hospital utilization patterns and costs for adult sickle cell patients in Illinois.

    PubMed Central

    Woods, K; Karrison, T; Koshy, M; Patel, A; Friedmann, P; Cassel, C

    1997-01-01

    OBJECTIVES: To determine population size, demographic characteristics, hospital utilization patterns, the specialties of physicians providing care, and costs for hospitalized adult sickle cell patients in Illinois. METHODS: A statewide, administrative dataset for the two-year period from january 1992 through December 1993 was analyzed retrospectively. RESULTS: There were 8403 admissions among 1189 individual sickle cell patients for the two-year period. Eighty-five percent of patients resided in the Chicago metropolitan area. The median age of the 1189 patients was 29; two-thirds had Medicaid or Medicare coverage. Emergency departments were the primary source of admissions (85.7%). The most common admitting diagnosis was painful crisis (97.4%), and average length of stay was four days. The median number of admissions per patient was three; most patients (85%) used only one or two hospitals. A small group used more than four hospitals and accounted for 23% of statewide admissions. Primary care physicians cared for most patients, and total hospitalization charges were more than $59 million. CONCLUSIONS: In Illinois the adult sickle cell population is concentrated in major urban centers, primarily the Chicago metropolitan area. These patients accounted for approximately 8400 admissions and more than $59 million in hospital charges during the two-year study period. A small group of patients used multiple hospitals and accounted for more than 23% of total hospitalization charges. This study shows the necessity of and provides a useful framework for developing targeted programs for adult sickle cell patients as well as for training physicians to efficiently provide comprehensive health care services for this population. PMID:9018288

  12. Differential utilization of binding loop flexibility in T cell receptor ligand selection and cross-reactivity

    PubMed Central

    Ayres, Cory M.; Scott, Daniel R.; Corcelli, Steven A.; Baker, Brian M.

    2016-01-01

    Complementarity determining region (CDR) loop flexibility has been suggested to play an important role in the selection and binding of ligands by T cell receptors (TCRs) of the cellular immune system. However, questions remain regarding the role of loop motion in TCR binding, and crystallographic structures have raised questions about the extent to which generalizations can be made. Here we studied the flexibility of two structurally well characterized αβ TCRs, A6 and DMF5. We found that the two receptors utilize loop motion very differently in ligand binding and cross-reactivity. While the loops of A6 move rapidly in an uncorrelated fashion, those of DMF5 are substantially less mobile. Accordingly, the mechanisms of binding and cross-reactivity are very different between the two TCRs: whereas A6 relies on conformational selection to select and bind different ligands, DMF5 uses a more rigid, permissive architecture with greater reliance on slower motions or induced-fit. In addition to binding site flexibility, we also explored whether ligand-binding resulted in common dynamical changes in A6 and DMF5 that could contribute to TCR triggering. Although binding-linked motional changes propagated throughout both receptors, no common features were observed, suggesting that changes in nanosecond-level TCR structural dynamics do not contribute to T cell signaling. PMID:27118724

  13. Titration of KATP channel expression in mammalian cells utilizing recombinant baculovirus transduction.

    PubMed

    Pfohl, Jeffrey L; Worley, Jennings F; Condreay, J Patrick; An, Gang; Apolito, Christopher J; Kost, Tom A; Truax, James F

    2002-01-01

    A variety of transfection approaches have been used to deliver plasmids encoding ion channel genes into cells. We have used the baculovirus transduction system, BacMam, to demonstrate transient expression of multi-subunit KATP channels in CHO-K1 and HEK-293 EBNA cells using sulfonylurea receptor 1 (SUR), SUR2A, SUR2B, and KIR 6.2 genes. [3H]-glyburide binding, patch clamp, and DiBAC4(3) measurements of membrane potential changes were used to monitor channel expression. BacMam delivery of each SUR isoform with KIR6.2 was demonstrated based on its pharmacological profiles. Expression levels of SUR1 and KIR6.2 were titrated by varying the viral concentration or time of virus addition, with functional activity measured in as little as 4-6 hours posttransduction. Further increases in BacMam virus induced sufficient KATP expression to dominate membrane potential without pharmacological opening of the channel. Independently altering treatment with virus containing either the SUR1 or KIR6.2 gene revealed interactions among subunits during formation of functional channels in the plasma membrane. This study demonstrates the utility and versatility of BacMam as a valuable gene delivery tool for the study of ion channel function.

  14. A 16 Month Survey of Cyclosporine Utilization Evaluation in Allogeneic Hematopoietic Stem Cell Transplant Recipients

    PubMed Central

    Tavakoli Ardakani, Maria; Tafazoli, Ali; Mehdizadeh, Mahshid; Hajifathali, Abbas; Dadashzadeh, Simin

    2016-01-01

    Objectives: Graft versus host disease (GVHD) is a life threatening reaction in the stem cell transplantation process. Nowadays Cyclosporine is the most commonly utilized agent for GVHD prophylaxis and it has a major role in successful transplantation. Cyclosporine has been applied for many years in this field but it could be stated that currently no general consensus is available for its optimal method of administration. Conditions related to cyclosporine administration and possible related adverse reactions observed closely in our patients with the aim of constructing a comprehensive practice guideline in the future. Patients and Methods: Allogeneic stem cell transplant recipients who have been taking cyclosporine were monitored during and after their hospitalization while recording all observations on predefined questionnaires on the basis of periodic clinical and laboratory examinations for a 16 month period. Results: Mean recorded duration of infusions was 1.44 ± 0.68 h and by twice daily administration, means intravenous and oral dose was 101.85 ± 22.03 mg and 219.28 ± 63.9 mg, respectively. A mean CsA trough level after about 12 h of specified unique doses was 223 ± 65 ng/mL. We found hypertension, nephrotoxicity, neurotoxicity, hypertension, and dyslipidemia in about 14, 20, 48, and 94 percent of patients. Conclusions: This study proposed that permanent guidance of healthcare team according to a fixed and standard method of cyclosporine administration routine with using efficient facilities and protocols would be helpful considerably for an optimal pharmacotherapy.

  15. A 16 Month Survey of Cyclosporine Utilization Evaluation in Allogeneic Hematopoietic Stem Cell Transplant Recipients

    PubMed Central

    Tavakoli Ardakani, Maria; Tafazoli, Ali; Mehdizadeh, Mahshid; Hajifathali, Abbas; Dadashzadeh, Simin

    2016-01-01

    Objectives: Graft versus host disease (GVHD) is a life threatening reaction in the stem cell transplantation process. Nowadays Cyclosporine is the most commonly utilized agent for GVHD prophylaxis and it has a major role in successful transplantation. Cyclosporine has been applied for many years in this field but it could be stated that currently no general consensus is available for its optimal method of administration. Conditions related to cyclosporine administration and possible related adverse reactions observed closely in our patients with the aim of constructing a comprehensive practice guideline in the future. Patients and Methods: Allogeneic stem cell transplant recipients who have been taking cyclosporine were monitored during and after their hospitalization while recording all observations on predefined questionnaires on the basis of periodic clinical and laboratory examinations for a 16 month period. Results: Mean recorded duration of infusions was 1.44 ± 0.68 h and by twice daily administration, means intravenous and oral dose was 101.85 ± 22.03 mg and 219.28 ± 63.9 mg, respectively. A mean CsA trough level after about 12 h of specified unique doses was 223 ± 65 ng/mL. We found hypertension, nephrotoxicity, neurotoxicity, hypertension, and dyslipidemia in about 14, 20, 48, and 94 percent of patients. Conclusions: This study proposed that permanent guidance of healthcare team according to a fixed and standard method of cyclosporine administration routine with using efficient facilities and protocols would be helpful considerably for an optimal pharmacotherapy. PMID:27610174

  16. Errors of utilizing carbon monoxide uptake to determine red cell mass

    SciTech Connect

    McNeil, J.; Mundie, T.; Ripple, G.; Jackson, B.; Martin, D. )

    1991-03-11

    Carbon monoxide (CO) has been used to approximate effective red cell mass (E{sub RCM}) utilizing the assumption that CO has a simple volume of distribution as carboxyhemoglobin (COHb). However, CO is known to bind to other heme molecules in vitro, and the extent of binding in vivo is unknown. E{sub RCM} was determined for six adult female splenectomized sheep using a CO containing re-breathing exposure system which allowed the animal to rapidly attain a steady state of COHb. Carbon dioxide was removed from the system and O{sub 2} was added as needed to maintain a constant system volume and O{sub 2} concentration. E{sub RCM} was calculated from measurements using a low, medium and high CO concentration. Results were compared with measurements of RCM done using the chromium 51 dilution method. The mean RCM ({plus minus} S.D.) was 1.87 {plus minus} 0.27 liters; while the mean E{sub RCM} ({plus minus} S.D.) was 2.56 {plus minus} 0.40. The E{sub RCM} was found to be 26.3% greater than the RCM suggesting significant tissue binding and that CO should not be used to estimate Red Cell Mass due to this binding.

  17. Circulating tumour cells: their utility in cancer management and predicting outcomes

    PubMed Central

    Krebs, Matthew G.; Hou, Jian-Mei; Ward, Tim H.; Blackhall, Fiona H.; Dive, Caroline

    2010-01-01

    Recent advances in technology now permit robust and reproducible detection of circulating tumour cells (CTCs) from a simple blood test. Standardization in methodology has been instrumental in facilitating multicentre trials with the purpose of evaluating the clinical utility of CTCs. We review the current body of evidence supporting the prognostic value of CTC enumeration in breast, prostate and colorectal cancer, using standardized approaches, and studies evaluating the correlation of CTC number with radiological outcome. The exploitation of CTCs in cancer management, however, is now extending beyond prognostication. As technologies emerge to characterize CTCs at the molecular level, biological information can be obtained in real time, with the promise of serving as a ‘surrogate tumour biopsy’. Current studies illuminate the potential of CTCs as pharmacodynamic and predictive biomarkers and potentially their use in revealing drug resistance in real time. Approaches for CTC characterization are summarized and the potential of CTCs in cancer patient management exemplified via the detection of epidermal growth factor receptor mutations from CTCs in patients with non-small cell lung cancer. The opportunity to learn more about the biology of metastasis through CTC analysis is now being realized with the horizon of CTC-guided development of novel anticancer therapies. PMID:21789147

  18. Utility of KRAS mutation detection using circulating cell-free DNA from patients with colorectal cancer.

    PubMed

    Yamada, Takeshi; Iwai, Takuma; Takahashi, Goro; Kan, Hayato; Koizumi, Michihiro; Matsuda, Akihisa; Shinji, Seiichi; Yamagishi, Aya; Yokoyama, Yasuyuki; Tatsuguchi, Atsushi; Kawagoe, Tatsuro; Kitano, Shiro; Nakayama, Masato; Matsumoto, Satoshi; Uchida, Eiji

    2016-07-01

    In this study, we evaluated the clinical utility of detecting KRAS mutations in circulating cell-free (ccf)DNA of metastatic colorectal cancer patients. We prospectively recruited 94 metastatic colorectal cancer patients. Circulating cell-free DNA was extracted from plasma samples and analyzed for the presence of seven KRAS point mutations. Using the Invader Plus assay with peptide nucleic acid clamping method and digital PCR, KRAS mutations were detected in the ccfDNA in 35 of 39 patients previously determined to have primary tumors containing KRAS mutations using the Luminex method, and in 5 of 55 patients with tumors containing wild-type KRAS. Curative resection was undertaken in 7 of 34 patients with primary and ccfDNA KRAS mutations, resulting in the disappearance of the mutation from the cell-free DNA in five of seven patients. Three of these patients had tumor recurrence and KRAS mutations in their ccfDNA reappeared. Epidermal growth factor receptor blockade was administered to 24 of the KRAS tumor wild-type patients. Of the 24 patients with wild-type KRAS in their primary tumors, three patients had KRAS mutations in their ccfDNA and did not respond to treatment with epidermal growth factor receptor (EGFR) blockade. We also detected a new KRAS mutation in five patients during chemotherapy with EGFR blockade, before disease progression was detectable with imaging. The detection of KRAS mutations in ccfDNA is an attractive approach for predicting both treatment response and acquired resistance to EGFR blockade, and for detecting disease recurrence. PMID:27116474

  19. Osteogenic cell cultures cannot utilize exogenous sources of synthetic polyphosphate for mineralization.

    PubMed

    Ariganello, Marianne B; Omelon, Sidney; Variola, Fabio; Wazen, Rima M; Moffatt, Pierre; Nanci, Antonio

    2014-12-01

    Phosphate is critical for mineralization and deficiencies in the regulation of free phosphate lead to disease. Inorganic polyphosphates (polyPs) may represent a physiological source of phosphate because they can be hydrolyzed by biological phosphatases. To investigate whether exogenous polyP could be utilized for mineral formation, mineralization was evaluated in two osteogenic cell lines, Saos-2 and MC3T3, expressing different levels of tissue non-specific alkaline phosphatase (tnALP). The role of tnALP was further explored by lentiviral-mediated overexpression in MC3T3 cells. When cells were cultured in the presence of three different phosphate sources, there was a strong mineralization response with β-glycerophosphate (βGP) and orthophosphate (Pi) but none of the cultures sustained mineralization in the presence of polyP (neither chain length 17-Pi nor 42-Pi). Even in the presence of mineralizing levels of phosphate, low concentrations of polyP (50 μM) were sufficient to inhibit mineral formation. Energy-dispersive X-ray spectroscopy confirmed the presence of apatite-like mineral deposits in MC3T3 cultures supplemented with βGP, but not in those with polyP. While von Kossa staining was consistent with the presence or absence of mineral, an unusual Alizarin staining was obtained in polyP-treated MC3T3 cultures. This staining pattern combined with low Ca:P ratios suggests the persistence of Ca-polyP complexes, even with high residual ALP activity. In conclusion, under standard culture conditions, exogenous polyP does not promote mineral deposition. This is not due to a lack of active ALP, and unless conditions that favor significant processing of polyP are achieved, its mineral inhibitory capacity predominates.

  20. Screening of aptamers specific to colorectal cancer cells and stem cells by utilizing On-chip Cell-SELEX

    PubMed Central

    Hung, Lien-Yu; Wang, Chih-Hung; Che, Yu-Jui; Fu, Chien-Yu; Chang, Hwan-You; Wang, Kuan; Lee, Gwo-Bin

    2015-01-01

    Colorectal cancer (CRC) is the most frequently diagnosed cancer around the world, causing about 700,000 deaths every year. It is clear now that a small fraction of CRC, named colorectal cancer stem cells (CSCs) exhibiting self-renewal and extensive proliferative activities, are hard to be eradicated. Unfortunately, highly specific biomarkers for colorectal CSC (CR-CSCs) are lacking that prohibits the development of effective therapeutic strategies. This study designed and manufactured a novel microfluidic system capable of performing a fully automated cell-based, systematic evolution of ligands by exponential enrichment (SELEX) process. Eight CR-CSC/CRC-specific aptamers were successfully selected using the microfluidic chip. Three of the aptamers showed high affinities towards their respective target cells with a dissociation constant of 27.4, 28.5 and 12.3 nM, which are comparable to that of antibodies. PMID:25999049

  1. Compact fuel cell system utilizing a combination of hydrogen storage materials for optimized performance.

    SciTech Connect

    Chan, Jennifer P.; Dedrick, Daniel E.; Gross, Karl J.; Ng, Greg L.

    2004-12-01

    An entirely new class of light-weight reversible hydrides was recently discovered (the Ti-doped alanates)[1]. These NaAIH{sub 4}-based materials have demonstrated reversible hydrogen storage capacities of up to 5 wt%, nearly 4 times the gravimetrically density of commercial metal hydrides. For this reason, they have been considered a breakthrough for hydrogen storage in fuel cell vehicles. This project is the first to publish the use of alanates for the generation of electrical power and the first demonstration of a hydride-fueled elevated-temperature PEM Fuel Cell. Because the kinetics of hydrogen uptake and release by the alanate improves with elevated temperatures, novel concepts were tested for the purpose of developing a highly efficient stand-alone power system. A major focus of this work was on the modeling, design, construction and testing of an integrated fuel cell stack and hydrogen storage system that eliminates the need of complicated heat transfer systems and media. After extensive modeling efforts, a proof-of-concept system was built that employs an integrated fuel cell stack and hydride beds that balancing the generation of fuel cell waste heat with the endothermic release of hydrogen from the alanates. Our demonstration unit was capable of greater than one hour of operation on a single charge of hydrogen from the integrated 173 gram alanate bed. In addition, composite hydride materials with synergistic reaction heats were evaluated and tested to enhance the operational performance of the alanates. The composites provide a unique opportunity to utilize the heat produced from hydriding classic metal hydrides to improve both absorption and desorption rates of the alanates. A particular focus of the mixed storage materials work was to balance the thermodynamics and kinetics of the hydrides for start-up conditions. Modeling of the sorption properties proved invaluable in evaluating the optimum composition of hydrides. The modeling efforts were followed

  2. Acetate is a superior substrate for microbial fuel cell initiation preceding bioethanol effluent utilization.

    PubMed

    Sun, Guotao; Thygesen, Anders; Meyer, Anne S

    2015-06-01

    This study assessed cell voltage development, electricity recovery, and microbial community composition in response to initial substrate including acetate, xylose, acetate/xylose 1:1 mixture (ace/xyl), and bioethanol effluent (BE) during microbial fuel cell (MFC) operation at 1000 Ω external resistance. The BE mainly contained 20.5 g/L xylose, 1.8 g/L arabinose, and 2.5 g/L propionic acid. The MFCs initially fed with acetate showed shorter initiation time (1 day), higher average cell voltage (634 ± 9 mV), and higher coulombic efficiency (31.5 ± 0.5 %) than those initially fed with ace/xyl or xylose. However, BE-initiated MFCs only generated 162 ± 1 mV. The acetate-initiated MFCs exhibited longer adaptation time (21 h) and lower cell voltage (645 ± 10 mV) when the substrate was switched to xylose, whereas substrate switching to BE produced the highest voltage (656 mV), maximum power density (362 ± 27 mW/m(2)), maximum current density (709 ± 27 mA/m(2)), and coulombic efficiency (25 ± 0.5 %) in the acetate-initiated MFCs. The microbial community in acetate-initiated MFCs was less diverse and contained more electrogenic bacteria (13.9 ± 0.4 %) including Geobacter sulfurreducens and Desulfuromonas acetexigen than the MFCs initially fed with ace/xyl, xylose, and BE. After switching the substrate to xylose and subsequently to BE, the microbial community in the acetate-initiated MFCs became more diverse, while no significant changes were observed in ace/xyl-, xylose-, and BE-initiated MFCs. The results showed that initial substrate affected the power generation and the capability to adapt to the substrate alteration in MFCs. Acetate-initiated MFCs showed best performance in utilizing BE. PMID:25794875

  3. Promoting remyelination: utilizing a viral model of demyelination to assess cell-based therapies

    PubMed Central

    Marro, Brett S; Blanc, Caroline A; Loring, Jeanne F; Cahalan, Michael D; Lane, Thomas E

    2014-01-01

    Multiple sclerosis (MS) is a chronic inflammatory disease of the CNS. While a broad range of therapeutics effectively reduce the incidence of focal white matter inflammation and plaque formation for patients with relapse-remitting forms of MS, a challenge within the field is to develop therapies that allow for axonal protection and remyelination. In the last decade, growing interest has focused on utilizing neural precursor cells (NPCs) to promote remyelination. To understand how NPCs function in chronic demyelinating environments, several excellent pre-clinical mouse models have been developed. One well accepted model is infection of susceptible mice with neurotropic variants of mouse hepatitis virus (MHV) that undergo chronic demyelination exhibiting clinical and histopathologic similarities to MS patients. Combined with the possibility that an environmental agent such as a virus could trigger MS, the MHV model of demyelination presents a relevant mouse model to assess the therapeutic potential of NPCs transplanted into an environment in which inflammatory-mediated demyelination is established. PMID:25245576

  4. Fabrication and Performance of Zirconia Electrolysis Cells for Carbon Dioxide Reduction for Mars In Situ Resource Utilization Applications

    NASA Technical Reports Server (NTRS)

    Minh, N. Q.; Chung, B. W.; Doshi, R.; Lear, G. R.; Montgomery, K.; Ong, E. T.

    1999-01-01

    The use of the Martian atmosphere (95% CO2) to produce oxygen (for propellant and life support) can significantly lower the required launch mass and dramatically reduce the total cost for Mars missions. Zirconia electrolysis cells are one of the technologies being considered for oxygen generation from carbon dioxide in Mars In Situ Resource Utilization (ISRU) production plants. The attractive features of the zirconia cell for this application include simple operation and lightweight, low volume system.

  5. Pancreatic polypeptide - a postulated new hormone: identification of its cellular storage site by light and electron microscopic immunocytochemistry.

    PubMed

    Larsson, L I; Sundler, F; Håkanson, R

    1976-07-01

    A peptide, referred to as pancreatic polypeptide (PP), has recently been isolated from the pancreas of chicken and of several mammals. PP is thought to be a pancreatic hormone. By the use of specific antisera we have demonstrated PP immunoreactivity in the pancreas of a number of mammals. The immunoreactivity was localized to a population of endocrine cells, distinct from the A, B and D cells. In most species the PP cells occurred in islets as well as in exocrine parenchyma; they often predominated in the pancreatic portion adjacent to the duodenum. In opossum and dog, PP cells were found also in the gastric mucosa. In opossum, the PP cells displayed formaldehyde - induced fluorescence typical of dopamine, whereas no formaldehyde-induced fluorescence was detected in the PP cells of mouse, rat and guinea pig. Also in these latter species, however, PP cells appear to possess amine-handling properties, a feature common to many peptide hormone-producing cells. The ultrastructure of the PP cells was defined by combining immunohistochemistry of semi-thin plastic sections with electron microscopy of adjacent ultrathin sections. PP cells show the ultrastructural features of peptide hormone-secreting cells. The PP cells of cat and dog contain fairly large, rather electron-lucent granules, and are probably identical with the previously described F cells. The PP cells of rat, guinea-pig, chinchilla and man contain small, fairly electron-dense granules. In these latter species no F cells are found. By immunoperoxidase staining of ultrathin sections, the PP immunoreactivity was found to be localized to the cytoplasmic granules. These observations provide support for the view that PP is a true pancreatic hormone. PMID:782992

  6. A model of the Capital Cost of a natural gas-fired fuel cell based Central Utilities Plant

    SciTech Connect

    Not Available

    1993-06-30

    This model defines the methods used to estimate the cost associated with acquisition and installation of capital equipment of the fuel cell systems defined by the central utility plant model. The capital cost model estimates the cost of acquiring and installing the fuel cell unit, and all auxiliary equipment such as a boiler, air conditioning, hot water storage, and pumps. The model provides a means to adjust initial cost estimates to consider learning associated with the projected level of production and installation of fuel cell systems. The capital cost estimate is an input to the cost of ownership analysis where it is combined with operating cost and revenue model estimates.

  7. A fiber optics system for monitoring utilization of ZnO adsorbent beds during desulfurization for logistic fuel cell applications

    NASA Astrophysics Data System (ADS)

    Sujan, Achintya; Yang, Hongyun; Dimick, Paul; Tatarchuk, Bruce J.

    2016-05-01

    An in-situ fiber optic based technique for direct measurement of capacity utilization of ZnO adsorbent beds by monitoring bed color changes during desulfurization for fuel cell systems is presented. Adsorbents composed of bulk metal oxides (ZnO) and supported metal oxides (ZnO/SiO2 and Cusbnd ZnO/SiO2) for H2S removal at 22 °C are examined. Adsorbent bed utilization at breakthrough is determined by the optical sensor as the maximum derivative of area under UV-vis spectrum from 250 to 800 nm observed as a function of service time. Since the response time of the sensor due to bed color change is close to bed breakthrough time, a series of probes along the bed predicts utilization of the portion of bed prior to H2S breakthrough. The efficacy of the optical sensor is evaluated as a function of inlet H2S concentration, H2S flow rate and desulfurization in presence of CO, CO2 and moisture in feed. A 6 mm optical probe is employed to measure utilization of a 3/16 inch ZnO extrudate bed for H2S removal. It is envisioned that with the application of the optical sensor, desulfurization can be carried out at high adsorbent utilization and low operational costs during on-board miniaturized fuel processing for logistic fuel cell power systems.

  8. Ultrastructural examination of the corticocollicular pathway in the guinea pig: a study using electron microscopy, neural tracers, and GABA immunocytochemistry.

    PubMed

    Nakamoto, Kyle T; Mellott, Jeffrey G; Killius, Jeanette; Storey-Workley, Megan E; Sowick, Colleen S; Schofield, Brett R

    2013-01-01

    Projections from auditory cortex (AC) can alter the responses of cells in the inferior colliculus (IC) to sounds. Most IC cells show excitation and inhibition after stimulation of the AC. AC axons release glutamate and excite their targets, so inhibition is presumed to result from cortical activation of GABAergic IC cells that inhibit other IC cells via local projections. However, it is not known whether cortical axons contact GABAergic IC cells directly. We labeled corticocollicular axons by injecting fluorescent dextrans into the AC in guinea pigs. We visualized the tracer with diaminobenzidine and processed the tissue for electron microscopy. We identified presumptive GABAergic profiles with post-embedding anti-GABA immunogold histochemistry on ultrathin sections. We identified dextran-labeled cortical boutons in the IC and identified their postsynaptic targets according to morphology (e.g., spine, dendrite) and GABA-reactivity. Cortical synapses were observed in all IC subdivisions, but were comparatively rare in the central nucleus. Cortical boutons contain round vesicles and few mitochondria. They form asymmetric synapses with spines (most frequently), dendritic shafts and, least often, with cell bodies. Excitatory boutons in the IC can be classified as large, medium or small; most cortical boutons belong to the small excitatory class, while a minority (~14%) belong to the medium excitatory class. Approximately 4% of the cortical targets were GABA-positive; these included dendritic shafts, spines, and cell bodies. We conclude that the majority of cortical boutons contact non-GABAergic (i.e., excitatory) IC cells and a small proportion (4%) contact GABAergic cells. Given that most IC cells show inhibition (as well as excitation) after cortical stimulation, it is likely that the majority of cortically-driven inhibition in the IC results from cortical activation of a relatively small number of IC GABAergic cells that have extensive local axons. PMID:23734104

  9. Utilization of cellulose microcapillary tubes as a model system for culturing and viral infection of mammalian cells.

    PubMed

    Venter, Eudri; Van Der Merwe, Christiaan F; Van Staden, Vida

    2012-10-01

    Cryofixation by high-pressure freezing (HPF) and freeze substitution (FS) gives excellent preservation of intracellular membranous structures, ideal for ultrastructural investigations of virus infected cells. Conventional sample preparation methods of tissue cultured cells can however disrupt the association between neighboring cells or of viruses with the plasma membrane, which impacts upon the effectiveness whereby virus release from cells can be studied. We established a system for virus infection and transmission electron microscopy preparation of mammalian cells that allowed optimal visualization of membrane release events. African horse sickness virus (AHSV) is a nonenveloped virus that employs two different release mechanisms from mammalian cells, i.e., lytic release through a disrupted plasma membrane and a nonlytic budding-type release. Cellulose microcapillary tubes were used as support layer for culturing Vero cells. The cells grew to a confluent monolayer along the inside of the tubes and could readily be infected with AHSV. Sections of the microcapillary tubes proved easy to manipulate during the HPF procedure, showed no distortion or compression, and yielded well preserved cells in their native state. There was ample cell surface area available for visualization, which allowed detection of both types of virus release at the plasma membrane at a significantly higher frequency than when utilizing other methods. The consecutive culturing, virus infection and processing of cells within microcapillary tubes therefore represent a novel model system for monitoring intracellular virus life cycle and membrane release events, specifically suited to viruses that do not grow to high titers in tissue culture. PMID:22865476

  10. Fabrication and Performance of Zirconia Electrolysis Cells for Cabon Dioxide Reduction for Mars In Situ Resource Utilization Applications

    NASA Technical Reports Server (NTRS)

    Minh, N. Q.; Chung, B. W.; Doshi, R.; Lear, G. R.; Montgomery, K.; Ong, E. T.

    1999-01-01

    Use of the Martian atmosphere (95% CO2) to produce oxygen (for propellant and life support) can significantly lower the required launch mass and dramatically reduce the total cost for Mars missions. Zirconia electrolysis cells are one of the technologies being considered for oxygen generation from carbon dioxide in Mars In Situ Resource Utilization (ISRU) production plants. The attractive features of the zirconia cell for this application include simple operation and lightweight, low volume system. A zirconia electrolysis cell is an all-solid state device, based on oxygen-ion conducting zirconia electrolytes, that electrochemically reduces carbon dioxide to oxygen and carbon monoxide. The cell consists of two porous electrodes (the anode and cathode) separated by a dense zirconia electrolyte. Typical zirconia cells contain an electrolyte layer which is 200 to 400 micrometer thick. The electrical conductivity requirement for the electrolyte necessitates an operating temperature of 9000 to 10000C. Recently, the fabrication of zirconia cells by the tape calendering has been evaluated. This fabrication process provides a simple means of making cells having very thin electrolytes (5 to 30 micrometers). Thin zirconia electrolytes reduce cell ohmic losses, permitting efficient operation at lower temperatures (8000C or below). Thus, tape-calendered cells provides not only the potential of low temperature operation but also the flexibility in operating temperatures. This paper describes the fabrication of zirconia cells by the tape calendering method and discusses the performance results obtained to date.

  11. Exploring transduction mechanisms of protein transduction domains (PTDs) in living cells utilizing single-quantum dot tracking (SQT) technology.

    PubMed

    Suzuki, Yasuhiro

    2012-01-01

    Specific protein domains known as protein transduction domains (PTDs) can permeate cell membranes and deliver proteins or bioactive materials into living cells. Various approaches have been applied for improving their transduction efficacy. It is, therefore, crucial to clarify the entry mechanisms and to identify the rate-limiting steps. Because of technical limitations for imaging PTD behavior on cells with conventional fluorescent-dyes, how PTDs enter the cells has been a topic of much debate. Utilizing quantum dots (QDs), we recently tracked the behavior of PTD that was derived from HIV-1 Tat (TatP) in living cells at the single-molecule level with 7-nm special precision. In this review article, we initially summarize the controversy on TatP entry mechanisms; thereafter, we will focus on our recent findings on single-TatP-QD tracking (SQT), to identify the major sequential steps of intracellular delivery in living cells and to discuss how SQT can easily provide direct information on TatP entry mechanisms. As a primer for SQT study, we also discuss the latest findings on single particle tracking of various molecules on the plasma membrane. Finally, we discuss the problems of QDs and the challenges for the future in utilizing currently available QD probes for SQT. In conclusion, direct identification of the rate-limiting steps of PTD entry with SQT should dramatically improve the methods for enhancing transduction efficiency. PMID:22368485

  12. Diagnostic utility of hepatocyte nuclear factor 1-beta immunoreactivity in endometrial carcinomas: lack of specificity for endometrial clear cell carcinoma.

    PubMed

    Fadare, Oluwole; Liang, Sharon X

    2012-12-01

    Hepatocyte nuclear factor 1-beta (HNF1β) has recently emerged as a relatively sensitive and specific marker for ovarian clear cell carcinoma. The purpose of this study is to assess the diagnostic utility of this marker for endometrial clear cell carcinoma. Immunohistochemical analysis was performed on 75 endometrial tissues using a goat polyclonal antibody raised against a peptide mapping at the C-terminus of human HNF1β protein. The 75 cases included 15 clear cell carcinomas, 20 endometrioid carcinomas, 15 endometrial serous carcinomas/uterine papillary serous carcinomas, 20 cases of normal endometrium, 2 cases of clear cell metaplasia, and 3 cases of Arias Stella reaction. Staining interpretations were based on a semiquantitative scoring system, a 0 to 12+ continuous numerical scale that was derived by multiplying the extent of staining (0 to 4+ scale) by the intensity of staining (0 to 3+ scale) for each case. HNF1β expression was found to be present in a wide spectrum of tissues. Twenty-seven (54%) of the 50 carcinomas displayed at least focal nuclear HNF1β expression, including 11 (73%) of 15, 9 (60%) of 15, and 7 (35%) of 20 clear cell, serous, and endometrioid carcinomas, respectively. The average nuclear staining scores for clear cell carcinomas, endometrioid carcinomas, and serous carcinomas were 5.2, 1.4, and 4.1, respectively. Clear cell carcinomas and endometrioid carcinomas displayed statistically significant differences regarding their nuclear staining scores (P = 0.0027), but clear cell carcinomas and endometrial serous carcinomas did not (P = 0.45). The calculated sensitivity of any nuclear HNF1β expression in classifying a carcinoma as being of the clear cell histotype was 73%, whereas the specificity was 54%. Nineteen of 20 normal endometrium samples displayed at least focal nuclear expression of HNF1β, and this expression was often diffuse. The 5 cases of benign histologic mimics of clear cell carcinomas (Arias Stella reaction and clear

  13. Disparities in utilization of autologous hematopoietic cell transplantation for treatment of multiple myeloma

    PubMed Central

    Costa, Luciano J.; Huang, Jia-Xing; Hari, Parameswaran N.

    2015-01-01

    Background AHCT is an established therapy for multiple myeloma (MM) with impact in quality of remissions and survival. We analyzed the role of race-ethnicity, sex and age-disparities in AHCT utilization in US. Methods We combined MM incidence derived from SEER-18 with transplant activity reported to the CIBMTR for the period of 2005–2009 to assess the impact of disparities in AHCT. Utilization (number of transplants/new cases) was compared between groups using relative utilization ratio (RUR), defined as [utilization for a given category]/[utilization for the entire population]. Results Data was obtained from 22,462 actual MM cases and 13,311 AHCT. Age-adjusted RUR was 1.17 (1.15–1.19) among non-Hispanic Whites (NHW), higher than in non-Hispanic Blacks (NHB) [0.69 (0.67–0.72), P<0.0002], Hispanics [0.64 (0.60–0.69), p<0.002] and Asians [0.65 (0.58–0.73), P<0.0002]. There was higher AHCT utilization in men than in women among Hispanics (age-adjusted RUR 0.72 vs. 0.56, P=0.007), but not among NHW, NHB or Asians. Sex disparity prevents 1.3% of potential AHCTs in MM (10.4% among Hispanics). Racial-ethnic disparities prevent 13.8% of AHCTs (44.7% in Hispanic and Asians, 39.9% in NHBs). Conclusions Race-ethnicity disparity greatly affects AHCT utilization in MM. Sex disparity plays a lesser role, except among Hispanics. The ongoing decrease in age disparity will continue to drive major increase of AHCT activity. A 2-year and 5-year increase in age of the AHCT population would result in respectively 12% and 32% increase in volume of AHCT. PMID:25555447

  14. Enhanced Electron Lifetimes in Dye-Sensitized Solar Cells Using a Dichromophoric Porphyrin: The Utility of Intermolecular Forces.

    PubMed

    Zhao, Long; Wagner, Pawel; van der Salm, Holly; Gordon, Keith C; Mori, Shogo; Mozer, Attila J

    2015-10-01

    Electron lifetimes in dye-sensitized solar cells employing a porphyrin dye, an organic dye, a 1:1 mixture of the two dyes, and a dichromophoric dye design consisting of the two dyes using a nonconjugated linker were measured, suggesting that the dispersion force of the organic dyes has a significant detrimental effect on the electron lifetime and that the dichromophoric design can be utilized to control the effect of the dispersion force.

  15. Understanding photoreceptor outer segment phagocytosis: use and utility of RPE cells in culture.

    PubMed

    Mazzoni, Francesca; Safa, Hussein; Finnemann, Silvia C

    2014-09-01

    RPE cells are the most actively phagocytic cells in the human body. In the eye, RPE cells face rod and cone photoreceptor outer segments at all times but contribute to shedding and clearance phagocytosis of distal outer segment tips only once a day. Analysis of RPE phagocytosis in situ has succeeded in identifying key players of the RPE phagocytic mechanism. Phagocytic processes comprise three distinct phases, recognition/binding, internalization, and digestion, each of which is regulated separately by phagocytes. Studies of phagocytosis by RPE cells in culture allow specifically analyzing and manipulating these distinct phases to identify their molecular mechanisms. Here, we compare similarities and differences of primary, immortalized, and stem cell-derived RPE cells in culture to RPE cells in situ with respect to phagocytic function. We discuss in particular potential pitfalls of RPE cell culture phagocytosis assays. Finally, we point out considerations for phagocytosis assay development for future studies.

  16. Variable Responses to Carbon Utilization between Planktonic and Biofilm Cells of a Human Carrier Strain of Salmonella enterica Serovar Typhi.

    PubMed

    Kalai Chelvam, Kalaivani; Yap, Kien Pong; Chai, Lay Ching; Thong, Kwai Lin

    2015-01-01

    Salmonella enterica serovar Typhi (S. Typhi) is a foodborne pathogen that causes typhoid fever and infects only humans. The ability of S. Typhi to survive outside the human host remains unclear, particularly in human carrier strains. In this study, we have investigated the catabolic activity of a human carrier S. Typhi strain in both planktonic and biofilm cells using the high-throughput Biolog Phenotype MicroArray, Minimum Biofilm Eradication Concentration (MBEC) biofilm inoculator (96-well peg lid) and whole genome sequence data. Additional strains of S. Typhi were tested to further validate the variation of catabolism in selected carbon substrates in the different bacterial growth phases. The analyzes of the carbon utilization data indicated that planktonic cells of the carrier strain, S. Typhi CR0044 could utilize a broader range of carbon substrates compared to biofilm cells. Pyruvic acid and succinic acid which are related to energy metabolism were actively catabolised in the planktonic stage compared to biofilm stage. On the other hand, glycerol, L-fucose, L-rhamnose (carbohydrates) and D-threonine (amino acid) were more actively catabolised by biofilm cells compared to planktonic cells. Notably, dextrin and pectin could induce strong biofilm formation in the human carrier strain of S. Typhi. However, pectin could not induce formation of biofilm in the other S. Typhi strains. Phenome data showed the utilization of certain carbon substrates which was supported by the presence of the catabolism-associated genes in S. Typhi CR0044. In conclusion, the findings showed the differential carbon utilization between planktonic and biofilm cells of a S. Typhi human carrier strain. The differences found in the carbon utilization profiles suggested that S. Typhi uses substrates mainly found in the human biliary mucus glycoprotein, gallbladder, liver and cortex of the kidney of the human host. The observed diversity in the carbon catabolism profiles among different S

  17. Clinical utility of natural killer cells in cancer therapy and transplantation

    PubMed Central

    Knorr, David; Bachanova, Veronika; Verneris, Michael R.; Miller, Jeffrey S.

    2014-01-01

    Natural killer (NK) cells recognize deranged cells that display stress receptors or loss of major histocompatibility complex (MHC) class I. During development, NK cells become “licensed” only after they encounter cognate human leucocyte antigen (HLA) class I, leading to the acquisition of effector function. NK cells can be exploited for cancer therapy in several ways. These include targeting within monoclonal antibodies alone or combined with ex vivo and in vivo NK cell activation to facilitate adoptive immunotherapy using donor-derived NK cell products to induce graft-vs-tumor effects. In the adoptive transfer setting, persistence and in vivo expansion requires lymphodepleting chemotherapy to prevent rejection and provide homeostatic cytokines (such as IL-15) that activate NK cells. IL-15 has the advantage of avoiding regulatory T-cell expansion. Clinical applications are currently being tested. To enhance in vivo expansion, IL-2 has been used at low doses. However, low dose administration also leads to the stimulation of regulatory T cells. Monoclonal antibodies and bispecific killer engagers (BiKEs) may enhance specificity by targeting CD16 on NK cells to tumor antigens. Inhibition of CD16 shedding may also promote enhanced cytotoxicity. Future strategies include exploiting favorable donor immunogenetics or ex vivo expansion of NK cells from blood, progenitors, or pluripotent cells. Comparative clinical trials are needed to test these approaches. PMID:24618042

  18. Utility of Induced Pluripotent Stem Cells for the Study and Treatment of Genetic Diseases: Focus on Childhood Neurological Disorders.

    PubMed

    Barral, Serena; Kurian, Manju A

    2016-01-01

    The study of neurological disorders often presents with significant challenges due to the inaccessibility of human neuronal cells for further investigation. Advances in cellular reprogramming techniques, have however provided a new source of human cells for laboratory-based research. Patient-derived induced pluripotent stem cells (iPSCs) can now be robustly differentiated into specific neural subtypes, including dopaminergic, inhibitory GABAergic, motorneurons and cortical neurons. These neurons can then be utilized for in vitro studies to elucidate molecular causes underpinning neurological disease. Although human iPSC-derived neuronal models are increasingly regarded as a useful tool in cell biology, there are a number of limitations, including the relatively early, fetal stage of differentiated cells and the mainly two dimensional, simple nature of the in vitro system. Furthermore, clonal variation is a well-described phenomenon in iPSC lines. In order to account for this, robust baseline data from multiple control lines is necessary to determine whether a particular gene defect leads to a specific cellular phenotype. Over the last few years patient-derived neural cells have proven very useful in addressing several mechanistic questions related to central nervous system diseases, including early-onset neurological disorders of childhood. Many studies report the clinical utility of human-derived neural cells for testing known drugs with repurposing potential, novel compounds and gene therapies, which then can be translated to clinical reality. iPSCs derived neural cells, therefore provide great promise and potential to gain insight into, and treat early-onset neurological disorders. PMID:27656126

  19. Utility of Induced Pluripotent Stem Cells for the Study and Treatment of Genetic Diseases: Focus on Childhood Neurological Disorders

    PubMed Central

    Barral, Serena; Kurian, Manju A.

    2016-01-01

    The study of neurological disorders often presents with significant challenges due to the inaccessibility of human neuronal cells for further investigation. Advances in cellular reprogramming techniques, have however provided a new source of human cells for laboratory-based research. Patient-derived induced pluripotent stem cells (iPSCs) can now be robustly differentiated into specific neural subtypes, including dopaminergic, inhibitory GABAergic, motorneurons and cortical neurons. These neurons can then be utilized for in vitro studies to elucidate molecular causes underpinning neurological disease. Although human iPSC-derived neuronal models are increasingly regarded as a useful tool in cell biology, there are a number of limitations, including the relatively early, fetal stage of differentiated cells and the mainly two dimensional, simple nature of the in vitro system. Furthermore, clonal variation is a well-described phenomenon in iPSC lines. In order to account for this, robust baseline data from multiple control lines is necessary to determine whether a particular gene defect leads to a specific cellular phenotype. Over the last few years patient-derived neural cells have proven very useful in addressing several mechanistic questions related to central nervous system diseases, including early-onset neurological disorders of childhood. Many studies report the clinical utility of human-derived neural cells for testing known drugs with repurposing potential, novel compounds and gene therapies, which then can be translated to clinical reality. iPSCs derived neural cells, therefore provide great promise and potential to gain insight into, and treat early-onset neurological disorders. PMID:27656126

  20. A model utilizing adult murine stem cells for creation of personalized islets for transplantation.

    PubMed

    Wang, J; Song, L J; Gerber, D A; Fair, J H; Rice, L; LaPaglia, M; Andreoni, K A

    2004-05-01

    Clinical islet cell transplantation has demonstrated great promise for diabetes treatment. Two major obstacles are the organ donor shortage and the immunoresponse. The purpose of this study was to create a model using the patient's own adult stem cell sources, possibly in combination with non-self cells, such as pancreatic, hepatic, or embryonic stem cells, to create "personalized" islets. We hypothesize that the reconstructed islets have the normal capability to produce insulin and glucagon with reduced immunoresponses after transplantation. Stem cells are a proliferating population of master cells that have the ability for self-renewal and multilineage differentiation. The recently developed photolithograph-based, biologic, microelectromechanic system (BioMEMS) technique supplies a useful tool for biomedical applications. Our lab has developed a novel method that integrates the adult stem cell and BioMEMS to reconstruct personalized islets. We selected islet-derived progenitor cells (IPC) for repairing and reconstructing STZ-diabetic islets. A6(+)/PYY(+) or A6(+)/ngn3(+) cells were selected to manipulate the neoislets. After 3 to 4 weeks in culture, the reconstructed cells formed islet-like clusters containing insulin or glucagon producing cells. The pilot results showed the ability of these reconstructed islets to correct hyperglycemia when transplanted into a STZ-diabetic isograft mouse model. Although several technical problems remain with the mouse model, namely, the difficulty to collect enough islets from a single mouse because of animal size, the mouse isograft model is suitable for personalized islet development.

  1. Utilization of GaN:Eu 3+ nanocrystals for the detection of programmed cell death

    NASA Astrophysics Data System (ADS)

    Bilyy, R.; Podhorodecki, A.; Nyk, M.; Stoika, R.; Zaichenko, A.; Zatryb, G.; Misiewicz, J.; Strek, W.

    2008-04-01

    In the current study we propose to use a new system for labeling biological processes. Gallium nitride nanocrystals doped by europium ions (nc-GaN:Eu 3+) have been obtained and used to identify the cells undergoing process of programmed cell death. Obtained by combustion method, GaN:Eu 3+ fluorescent nanocrystals have been covered with the polymeric envelope, bearing epoxy groups. Carbohydrate-binding protein-lectin-specifically recognizing cells undergoing programmed cell death was conjugated to the envelope of nanoparticles. Incubation of alive and dead cells with nanoparticles suspension and subsequent analysis using fluorescent and phase-contrast microscopy revealed predominate binding of nanoparticles to dead cells, while intact cell did not bind nanoparticles under the same conditions.

  2. Numerical analysis of acoustic impedance microscope utilizing acoustic lens transducer to examine cultured cells.

    PubMed

    Gunawan, Agus Indra; Hozumi, Naohiro; Takahashi, Kenta; Yoshida, Sachiko; Saijo, Yoshifumi; Kobayashi, Kazuto; Yamamoto, Seiji

    2015-12-01

    A new technique is proposed for non-contact quantitative cell observation using focused ultrasonic waves. This technique interprets acoustic reflection intensity into the characteristic acoustic impedance of the biological cell. The cells are cultured on a plastic film substrate. A focused acoustic beam is transmitted through the substrate to its interface with the cell. A two-dimensional (2-D) reflection intensity profile is obtained by scanning the focal point along the interface. A reference substance is observed under the same conditions. These two reflections are compared and interpreted into the characteristic acoustic impedance of the cell based on a calibration curve that was created prior to the observation. To create the calibration curve, a numerical analysis of the sound field is performed using Fourier Transforms and is verified using several saline solutions. Because the cells are suspended by two plastic films, no contamination is introduced during the observation. In a practical observation, a sapphire lens transducer with a center frequency of 300 MHz was employed using ZnO thin film. The objects studied were co-cultured rat-derived glial (astrocyte) cells and glioma cells. The result was the clear observation of the internal structure of the cells. The acoustic impedance of the cells was spreading between 1.62 and 1.72 MNs/m(3). Cytoskeleton was indicated by high acoustic impedance. The introduction of cytochalasin-B led to a significant reduction in the acoustic impedance of the glioma cells; its effect on the glial cells was less significant. It is believed that this non-contact observation method will be useful for continuous cell inspections.

  3. Aspartate Rescues S-phase Arrest Caused by Suppression of Glutamine Utilization in KRas-driven Cancer Cells.

    PubMed

    Patel, Deven; Menon, Deepak; Bernfeld, Elyssa; Mroz, Victoria; Kalan, Sampada; Loayza, Diego; Foster, David A

    2016-04-22

    During G1-phase of the cell cycle, normal cells respond first to growth factors that indicate that it is appropriate to divide and then later in G1 to the presence of nutrients that indicate sufficient raw material to generate two daughter cells. Dividing cells rely on the "conditionally essential" amino acid glutamine (Q) as an anaplerotic carbon source for TCA cycle intermediates and as a nitrogen source for nucleotide biosynthesis. We previously reported that while non-transformed cells arrest in the latter portion of G1 upon Q deprivation, mutant KRas-driven cancer cells bypass the G1 checkpoint, and instead, arrest in S-phase. In this study, we report that the arrest of KRas-driven cancer cells in S-phase upon Q deprivation is due to the lack of deoxynucleotides needed for DNA synthesis. The lack of deoxynucleotides causes replicative stress leading to activation of the ataxia telangiectasia and Rad3-related protein (ATR)-mediated DNA damage pathway, which arrests cells in S-phase. The key metabolite generated from Q utilization was aspartate, which is generated from a transaminase reaction whereby Q-derived glutamate is converted to α-ketoglutarate with the concomitant conversion of oxaloacetate to aspartate. Aspartate is a critical metabolite for both purine and pyrimidine nucleotide biosynthesis. This study identifies the molecular basis for the S-phase arrest caused by Q deprivation in KRas-driven cancer cells that arrest in S-phase in response to Q deprivation. Given that arresting cells in S-phase sensitizes cells to apoptotic insult, this study suggests novel therapeutic approaches to KRas-driven cancers.

  4. Fuel cell and system for supplying electrolyte thereto utilizing cascade feed

    DOEpatents

    Feigenbaum, Haim

    1984-01-01

    An electrolyte distribution supply system for use with a fuel cell having a wicking medium for drawing electrolyte therein is formed by a set of containers of electrolyte joined to respective fuel cells or groups thereof in a stack of such cells. The electrolyte is separately stored so as to provide for electrical isolation between electrolytes of the individual cells or groups of cells of the stack. Individual storage compartments are coupled by individual tubes, the ends of the respective tubes terminating on the wicking medium in each of the respective fuel cells. The individual compartments are filled with electrolyte by allowing the compartments to overflow such as in a cascading fashion thereby maintaining the requisite depth of electrolyte in each of the storage compartments. The individual compartments can also contain packed carbon fibers to provide a three stage electrolyte distribution system.

  5. Armored CAR T-cells: utilizing cytokines and pro-inflammatory ligands to enhance CAR T-cell anti-tumour efficacy.

    PubMed

    Yeku, Oladapo O; Brentjens, Renier J

    2016-04-15

    Chimaeric antigen receptor (CAR) T-cells are T-cells that have been genetically modified to express an artificial construct consisting of a synthetic T-cell receptor (TCR) targeted to a predetermined antigen expressed on a tumour. Coupling the T-cell receptor to a CD3ζ signalling domain paved the way for first generation CAR T-cells that were efficacious against cluster of differentiation (CD)19-expressing B-cell malignancies. Optimization with additional signalling domains such as CD28 or 4-1BB in addition to CD3ζ provided T-cell activation signal 2 and further improved the efficacy and persistence of these second generation CAR T-cells. Third generation CAR T-cells which utilize two tandem costimulatory domains have also been reported. In this review, we discuss a different approach to optimization of CAR T-cells. Through additional genetic modifications, these resultant armored CAR T-cells are typically modified second generation CAR T-cells that have been further optimized to inducibly or constitutively secrete active cytokines or express ligands that further armor CAR T-cells to improve efficacy and persistence. The choice of the 'armor' agent is based on knowledge of the tumour microenvironment and the roles of other elements of the innate and adaptive immune system. Although there are several variants of armored CAR T-cells under investigation, here we focus on three unique approaches using interleukin-12 (IL-12), CD40L and 4-1BBL. These agents have been shown to further enhance CAR T-cell efficacy and persistence in the face of a hostile tumour microenvironment via different mechanisms.

  6. The establishment of a production-ready manufacturing process utilizing thin silicon substrates for solar cells

    NASA Technical Reports Server (NTRS)

    Pryor, R. A.

    1980-01-01

    Three inch diameter Czochralski silicon substrates sliced directly to 5 mil, 8 mil, and 27 mil thicknesses with wire saw techniques were procured. Processing sequences incorporating either diffusion or ion implantation technologies were employed to produce n+p or n+pp+ solar cell structures. These cells were evaluated for performance, ease of fabrication, and cost effectiveness. It was determined that the use of 7 mil or even 4 mil wafers would provide near term cost reductions for solar cell manufacturers.

  7. Effect of Black Soybean Koji Extract on Glucose Utilization and Adipocyte Differentiation in 3T3-L1 Cells

    PubMed Central

    Huang, Chi-Chang; Huang, Wen-Ching; Hou, Chien-Wen; Chi, Yu-Wei; Huang, Hui-Yu

    2014-01-01

    Adipocyte differentiation and the extent of subsequent fat accumulation are closely related to the occurrence and progression of diseases such as insulin resistance and obesity. Black soybean koji (BSK) is produced by the fermentation of black soybean with Aspergilllus awamori. Previous study indicated that BSK extract has antioxidative and multifunctional bioactivities, however, the role of BSK in the regulation of energy metabolism is still unclear. We aimed to investigate the effect of glucose utilization on insulin-resistant 3T3-L1 preadipocytes and adipogenesis-related protein expression in differentiated adipocytes with BSK treatment. Cytoxicity assay revealed that BSK did not adversely affect cell viability at levels up to 200 μg/mL. The potential for glucose utilization was increased by increased glucose transporter 1 (GLUT1), GLUT4 and protein kinase B (AKT) protein expression in insulin-resistant 3T3-L1 cells in response to BSK treatment. Simultaneously, BSK inhibited lipid droplet accumulation in differentiated 3T3-L1 cells. The inhibitory effect of adipogenesis was associated with downregulated peroxisome proliferator-activated receptor γ (PPARγ) level and upregulated Acrp30 protein expression. Our results suggest that BSK extract could improve glucose uptake by modulating GLUT1 and GLUT4 expression in a 3T3-L1 insulin-resistance cell model. In addition, BSK suppressed differentiation and lipid accumulation in mature 3T3-L1 adipocytes, which may suggest its potential for food supplementation to prevent obesity and related metabolic abnormalities. PMID:24821545

  8. Efficient immobilization of mushroom tyrosinase utilizing whole cells from Agaricus bisporus and its application for degradation of bisphenol A.

    PubMed

    Kampmann, Markus; Boll, Stefan; Kossuch, Jan; Bielecki, Julia; Uhl, Stefan; Kleiner, Beatrice; Wichmann, Rolf

    2014-06-15

    A simple and efficient procedure for preparation and immobilization of tyrosinase enzyme was developed utilizing whole cells from the edible mushroom Agaricus bisporus, without the need for enzyme purification. Tyrosinase activity in the cell preparation remained constant during storage at 21 °C for at least six months. The cells were entrapped in chitosan and alginate matrix capsules and characterized with respect to their resulting tyrosinase activity. A modification of the alginate with colloidal silica enhanced the activity due to retention of both cells and tyrosinase from fractured cells, which otherwise leached from matrix capsules. The observed activity was similar to the activity that was obtained with immobilized isolated tyrosinase in the same material. Mushroom cells in water were susceptible to rapid inactivation, whereas the immobilized cells maintained 73% of their initial activity after 30 days of storage in water. Application in repeated batch experiments resulted in almost 100% conversion of endocrine disrupting bisphenol A (BPA) for 11 days, under stirring conditions, and 50-60% conversion after 20 days, without stirring under continuous usage. The results represent the longest yet reported application of immobilized tyrosinase for degradation of BPA in environmental water samples.

  9. Robust optimization of a mathematical model to design a dynamic cell formation problem considering labor utilization

    NASA Astrophysics Data System (ADS)

    Vafaeinezhad, Moghadaseh; Kia, Reza; Shahnazari-Shahrezaei, Parisa

    2016-11-01

    Cell formation (CF) problem is one of the most important decision problems in designing a cellular manufacturing system includes grouping machines into machine cells and parts into part families. Several factors should be considered in a cell formation problem. In this work, robust optimization of a mathematical model of a dynamic cell formation problem integrating CF, production planning and worker assignment is implemented with uncertain scenario-based data. The robust approach is used to reduce the effects of fluctuations of the uncertain parameters with regards to all possible future scenarios. In this research, miscellaneous cost parameters of the cell formation and demand fluctuations are subject to uncertainty and a mixed-integer nonlinear programming model is developed to formulate the related robust dynamic cell formation problem. The objective function seeks to minimize total costs including machine constant, machine procurement, machine relocation, machine operation, inter-cell and intra-cell movement, overtime, shifting labors between cells and inventory holding. Finally, a case study is carried out to display the robustness and effectiveness of the proposed model. The tradeoff between solution robustness and model robustness is also analyzed in the obtained results.

  10. The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells

    PubMed Central

    Rao, Sudheendra; Morales, Alejo A.; Pearse, Damien D.

    2015-01-01

    The introduction of genes into glial cells for mechanistic studies of cell function and as a therapeutic for gene delivery is an expanding field. Though viral vector based systems do exhibit good delivery efficiency and long-term production of the transgene, the need for transient gene expression, broad and rapid gene setup methodologies, and safety concerns regarding in vivo application still incentivize research into the use of nonviral gene delivery methods. In the current study, aviral gene delivery vectors based upon cationic lipid (Lipofectamine 3000) lipoplex or polyethylenimine (Viromer RED) polyplex technologies were examined in cell lines and primary glial cells for their transfection efficiencies, gene expression levels, and toxicity. The transfection efficiencies of polyplex and lipoplex agents were found to be comparable in a limited, yet similar, transfection setting, with or without serum across a number of cell types. However, differential effects on cell-specific transgene expression and reduced viability with cargo loaded polyplex were observed. Overall, our data suggests that polyplex technology could perform comparably to the market dominant lipoplex technology in transfecting various cells lines including glial cells but also stress a need for further refinement of polyplex reagents to minimize their effects on cell viability. PMID:26539498

  11. Utility-Scale Power Router: Dynamic Control of Grid Assets Using Direct AC Converter Cells

    SciTech Connect

    2010-09-01

    ADEPT Project: Georgia Tech is developing a cost-effective, utility-scale power router that uses an enhanced transformer to more efficiently direct power on the grid. Existing power routing technologies are too expensive for widespread use, but the ability to route grid power to match real-time demand and power outages would significantly reduce energy costs for utilities, municipalities, and consumers. Georgia Tech is adding a power converter to an existing grid transformer to better control power flows at about 1/10th the cost of existing power routing solutions. Transformers convert the high-voltage electricity that is transmitted through the grid into the low-voltage electricity that is used by homes and businesses. The added converter uses fewer steps to convert some types of power and eliminates unnecessary power storage, among other improvements. The enhanced transformer is more efficient, and it would still work even if the converter fails, ensuring grid reliability.

  12. Differentiation-Dependent Energy Production and Metabolite Utilization: A Comparative Study on Neural Stem Cells, Neurons, and Astrocytes.

    PubMed

    Jády, Attila Gy; Nagy, Ádám M; Kőhidi, Tímea; Ferenczi, Szilamér; Tretter, László; Madarász, Emília

    2016-07-01

    While it is evident that the metabolic machinery of stem cells should be fairly different from that of differentiated neurons, the basic energy production pathways in neural stem cells (NSCs) or in neurons are far from clear. Using the model of in vitro neuron production by NE-4C NSCs, this study focused on the metabolic changes taking place during the in vitro neuronal differentiation. O2 consumption, H(+) production, and metabolic responses to single metabolites were measured in cultures of NSCs and in their neuronal derivatives, as well as in primary neuronal and astroglial cultures. In metabolite-free solutions, NSCs consumed little O2 and displayed a higher level of mitochondrial proton leak than neurons. In stem cells, glycolysis was the main source of energy for the survival of a 2.5-h period of metabolite deprivation. In contrast, stem cell-derived or primary neurons sustained a high-level oxidative phosphorylation during metabolite deprivation, indicating the consumption of own cellular material for energy production. The stem cells increased O2 consumption and mitochondrial ATP production in response to single metabolites (with the exception of glucose), showing rapid adaptation of the metabolic machinery to the available resources. In contrast, single metabolites did not increase the O2 consumption of neurons or astrocytes. In "starving" neurons, neither lactate nor pyruvate was utilized for mitochondrial ATP production. Gene expression studies also suggested that aerobic glycolysis and rapid metabolic adaptation characterize the NE-4C NSCs, while autophagy and alternative glucose utilization play important roles in the metabolism of stem cell-derived neurons.

  13. Differentiation-Dependent Energy Production and Metabolite Utilization: A Comparative Study on Neural Stem Cells, Neurons, and Astrocytes

    PubMed Central

    Jády, Attila Gy.; Nagy, Ádám M.; Kőhidi, Tímea; Ferenczi, Szilamér; Tretter, László

    2016-01-01

    While it is evident that the metabolic machinery of stem cells should be fairly different from that of differentiated neurons, the basic energy production pathways in neural stem cells (NSCs) or in neurons are far from clear. Using the model of in vitro neuron production by NE-4C NSCs, this study focused on the metabolic changes taking place during the in vitro neuronal differentiation. O2 consumption, H+ production, and metabolic responses to single metabolites were measured in cultures of NSCs and in their neuronal derivatives, as well as in primary neuronal and astroglial cultures. In metabolite-free solutions, NSCs consumed little O2 and displayed a higher level of mitochondrial proton leak than neurons. In stem cells, glycolysis was the main source of energy for the survival of a 2.5-h period of metabolite deprivation. In contrast, stem cell-derived or primary neurons sustained a high-level oxidative phosphorylation during metabolite deprivation, indicating the consumption of own cellular material for energy production. The stem cells increased O2 consumption and mitochondrial ATP production in response to single metabolites (with the exception of glucose), showing rapid adaptation of the metabolic machinery to the available resources. In contrast, single metabolites did not increase the O2 consumption of neurons or astrocytes. In “starving” neurons, neither lactate nor pyruvate was utilized for mitochondrial ATP production. Gene expression studies also suggested that aerobic glycolysis and rapid metabolic adaptation characterize the NE-4C NSCs, while autophagy and alternative glucose utilization play important roles in the metabolism of stem cell-derived neurons. PMID:27116891

  14. How Shigella Utilizes Ca(2+) Jagged Edge Signals during Invasion of Epithelial Cells.

    PubMed

    Bonnet, Mariette; Tran Van Nhieu, Guy

    2016-01-01

    Shigella, the causative agent of bacillary dysentery invades intestinal epithelial cells using a type III secretion system (T3SS). Through the injection of type III effectors, Shigella manipulates the actin cytoskeleton to induce its internalization in epithelial cells. At early invasion stages, Shigella induces atypical Ca(2+) responses confined at entry sites allowing local cytoskeletal remodeling for bacteria engulfment. Global Ca(2+) increase in the cell triggers the opening of connexin hemichannels at the plasma membrane that releases ATP in the extracellular milieu, favoring Shigella invasion and spreading through purinergic receptor signaling. During intracellular replication, Shigella regulates inflammatory and death pathways to disseminate within the epithelium. At later stages of infection, Shigella downregulates hemichannel opening and the release of extracellular ATP to dampen inflammatory signals. To avoid premature cell death, Shigella activates cell survival by upregulating the PI3K/Akt pathway and downregulating the levels of p53. Furthermore, Shigella interferes with pro-apoptotic caspases, and orients infected cells toward a slow necrotic cell death linked to mitochondrial Ca(2+) overload. In this review, we will focus on the role of Ca(2+) responses and their regulation by Shigella during the different stages of bacterial infection. PMID:26904514

  15. Functionality and antidiabetic utility of β- and L-cell containing pseudoislets.

    PubMed

    Green, Alastair D; Vasu, Srividya; Flatt, Peter R

    2016-06-10

    Unavailability of tissue and poor engraftment remain significant obstacles to clinical islet transplantation. Here, the therapeutic potential of pseudoislets generated from the insulin and GLP-1 releasing cell-lines MIN6 and GLUTag was investigated. Glucose and other secretagogues evoked 1.3-5.7 fold increases in insulin secretion from both pseudoislet types. Secretion expressed in relation to basal values did not greatly differ between configurations. Exposure of both types of pseudoislets to ninhydrin, H2O2, streptozotocin or cytokine cocktails decreased viability and increased apoptosis. However, combined pseudoislets exhibited enhanced resistance (1.2-1.7 fold increased LD50, 1.2-1.4 fold decreased apoptosis). Implantation of pseudoislets into streptozotocin-diabetic SCID mice precipitated cell masses containing immunoreactive insulin and GLP-1. Implantation of both pseudoislet types was associated with significant reductions in blood glucose, increased plasma insulin, greater bodyweight, decreased polydipsia and improved glucose tolerance. These changes greatly exaggerated in MIN6 pseudoislet recipients, with mice becoming severely hypoglycaemic. In contract, combined pseudoislet recipients achieved tempered restoration of normoglycaemia and exhibited increased plasma GLP-1, decreased plasma and pancreatic glucagon, increased pancreatic insulin and enhancements in islet β:α cells and the ratio of Ki67: TUNEL positive β-cells. MIN6 pseudoislet implantation increased islet β:α cell ratio but did not affect β-cell proliferation or hormone content. Our observations highlight the potential of combining insulin and GLP-1 cell therapy using heterotypic pseudoislets. PMID:27068377

  16. Utilization of physiological and taxonomic fluorescent probes to study Lactobacilli cells and response to pH challenge.

    PubMed

    Olszewska, Magdalena A; Kocot, Aleksandra M; Nynca, Anna; Łaniewska-Trokenheim, Łucja

    2016-11-01

    pH stress is recognized as an important feature for Lactobacillus in relation to lifestyle and commercial utility. Hence, this study aims to investigate the cell function of Lactobacilli cells subjected to pHs between 7.0 and 2.0. For this purpose, the Lactobacilli isolates of vegetable origin were first hybridized with fluorescent oligonucleotide rRNA probes for detecting Lactobacillus species. Then, cells were exposed to pH stress and labelled with fluorescent probes, carboxyfluorescein diacetate (CFDA) and propidium iodine (PI), which provided the insight into esterase activity and membrane integrity of cells. Among isolates, fluorescence in situ hybridization (FISH) enabled us to specifically detect L. plantarum and L. brevis. Interestingly, FCM analysis revealed that at pHs between 7.0 and 4.0 the cell membrane was intact, while after the exposure at pH 3.0, and 2.0 became perturbed or impaired. Finally, L. brevis and L. plantarum differed from each other in fluorescence labeling behaviour and culturability. However, the results showed that the same standard protocol for labeling enables discrimination of subpopulations of tested species. Depending on the species, the substantial culturability loss was observed at pH 3.0 and 2.0. These results suggest that the taxonomic and physiological fluorescent probes could be suitable for in situ detection of specific bacteria and rapid assessment of the physiological status of cells. PMID:27664742

  17. Utilization of physiological and taxonomic fluorescent probes to study Lactobacilli cells and response to pH challenge.

    PubMed

    Olszewska, Magdalena A; Kocot, Aleksandra M; Nynca, Anna; Łaniewska-Trokenheim, Łucja

    2016-11-01

    pH stress is recognized as an important feature for Lactobacillus in relation to lifestyle and commercial utility. Hence, this study aims to investigate the cell function of Lactobacilli cells subjected to pHs between 7.0 and 2.0. For this purpose, the Lactobacilli isolates of vegetable origin were first hybridized with fluorescent oligonucleotide rRNA probes for detecting Lactobacillus species. Then, cells were exposed to pH stress and labelled with fluorescent probes, carboxyfluorescein diacetate (CFDA) and propidium iodine (PI), which provided the insight into esterase activity and membrane integrity of cells. Among isolates, fluorescence in situ hybridization (FISH) enabled us to specifically detect L. plantarum and L. brevis. Interestingly, FCM analysis revealed that at pHs between 7.0 and 4.0 the cell membrane was intact, while after the exposure at pH 3.0, and 2.0 became perturbed or impaired. Finally, L. brevis and L. plantarum differed from each other in fluorescence labeling behaviour and culturability. However, the results showed that the same standard protocol for labeling enables discrimination of subpopulations of tested species. Depending on the species, the substantial culturability loss was observed at pH 3.0 and 2.0. These results suggest that the taxonomic and physiological fluorescent probes could be suitable for in situ detection of specific bacteria and rapid assessment of the physiological status of cells.

  18. Measuring relative utilization of aerobic glycolysis in breast cancer cells by positional isotopic discrimination.

    PubMed

    Yang, Da-Qing; Freund, Dana M; Harris, Benjamin R E; Wang, Defeng; Cleary, Margot P; Hegeman, Adrian D

    2016-09-01

    The ability of cancer cells to produce lactate through aerobic glycolysis is a hallmark of cancer. In this study, we established a positional isotopic labeling and LC-MS-based method that can specifically measure the conversion of glucose to lactate in glycolysis. We show that the rate of aerobic glycolysis is closely correlated with glucose uptake and lactate production in breast cancer cells. We also found that the production of [3-(13) C]lactate is significantly elevated in metastatic breast cancer cells and in early stage metastatic mammary tumors in mice. Our findings may enable the development of a biomarker for the diagnosis of aggressive breast cancer. PMID:27531463

  19. Light-splitting photovoltaic system utilizing two dual-junction solar cells

    SciTech Connect

    Xiong, Kanglin; Yang, Hui; Lu, Shulong; Dong, Jianrong; Zhou, Taofei; Wang, Rongxin; Jiang, Desheng

    2010-12-15

    There are many difficulties limiting the further development of monolithic multi-junction solar cells, such as the growth of lattice-mismatched material and the current matching constraint. As an alternative approach, the light-splitting photovoltaic system is investigated intensively in different aspects, including the energy loss mechanism and the choice of energy bandgaps of solar cells. Based on the investigation, a two-dual junction system has been implemented employing lattice-matched GaInP/GaAs and InGaAsP/InGaAs cells grown epitaxially on GaAs and InP substrates, respectively. (author)

  20. Alterations in cyclin-dependent protein kinase 5 (CDK5) protein levels, activity and immunocytochemistry in canine motor neuron disease.

    PubMed

    Green, S L; Vulliet, P R; Pinter, M J; Cork, L C

    1998-11-01

    Hereditary canine spinal muscular atrophy (HCSMA) is a dominantly inherited motor neuron disease in Brittany spaniels that is clinically characterized by progressive muscle weakness leading to paralysis. Histopathologically, degeneration is confined to motor neurons with accumulation of phosphorylated neurofilaments in axonal internodes. Cyclin-dependent kinase 5 (CDK5), a kinase related to the cell cycle kinase cdc2, phosphorylates neurofilaments and regulates neurofilament dynamics. We examined CDK5 activity, protein levels, and cellular immunoreactivity in nervous tissue from dogs with HCSMA, from closely age-matched controls and from dogs with other neurological diseases. On immunoblot analysis, CDK5 protein levels were increased in the HCSMA dogs (by approximately 1.5-fold in both the cytosolic and the particulate fractions). CDK5 activity was significantly increased (by approximately 3-fold) in the particulate fractions in the HCSMA dogs compared to all controls. The finding that CDK5 activity was increased in the young HCSMA homozygotes with the accelerated form of the disease, who do not show axonal swellings histologically, suggests that alterations in CDK5 occurs early in the pathogenesis, prior to the development of significant neurofilament pathology. Immunocytochemically, there was strong CDK5 staining of the nuclei, cytoplasm and axonal processes of the motor neurons in both control dogs and dogs with HCSMA. Further immunocytochemical studies demonstrated CDK5 staining where neurofilaments accumulated, in axonal swellings in the dogs with HCSMA. Our observations suggest phosphorylation-dependent events mediated by CDK5 occur in canine motor neuron disease.

  1. Epigallocatechin-3-gallate directly suppresses T cell proliferation through impaired IL-2 utilization and cell cycle progression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Epigallocatechin-3-gallate (EGCG), a bioactive component of green tea, has a variety of health impact. Previously we demonstrated that in vitro EGCG supplementation inhibited T cell response in mouse spleen cells. In the present study, we first extended our in vitro observation to in vivo and confir...

  2. Evolution of oxygen utilization in multicellular organisms and implications for cell signalling in tissue engineering

    PubMed Central

    Stamati, Katerina; Mudera, Vivek

    2011-01-01

    Oxygen is one of the critically defining elements resulting in the existence of eukaryotic life on this planet. The rise and fall of this element can be tracked through time and corresponds with the evolution of diverse life forms, development of efficient energy production (oxidative phosphorylation) in single cell organisms, the evolution of multicellular organisms and the regulation of complex cell phenotypes. By understanding these events, we can plot the effect of oxygen on evolution and its direct influence on different forms of life today, from the whole organism to specific cells within multicellular organisms. In the emerging field of tissue engineering, understanding the role of different levels of oxygen for normal cell function as well as control of complex signalling cascades is paramount to effectively build 3D tissues in vitro and their subsequent survival when implanted. PMID:22292107

  3. Assessing the utility of bipolar membranes for use in photoelectrochemical water-splitting cells.

    PubMed

    Vargas-Barbosa, Nella M; Geise, Geoffrey M; Hickner, Michael A; Mallouk, Thomas E

    2014-11-01

    Membranes are important in water-splitting solar cells because they prevent crossover of hydrogen and oxygen. Here, bipolar membranes (BPMs) were tested as separators in water electrolysis cells. Steady-state membrane and solution resistances, electrode overpotentials, and pH gradients were measured at current densities relevant to solar photoelectrolysis. Under forward bias conditions, electrodialysis of phosphate buffer ions creates a pH gradient across a BPM. Under reverse bias, the BPM can maintain a constant buffer pH on both sides of the cell, but a large membrane potential develops. Thus, the BPM does not present a viable solution for electrolysis in buffered electrolytes. However, the membrane potential is minimized when the anode and cathode compartments of the cell contain strongly basic and acidic electrolytes, respectively.

  4. Enhancing cellulose utilization for fuels and chemicals by genetic modification of plant cell wall architecture.

    PubMed

    Vermerris, Wilfred; Abril, Alejandra

    2015-04-01

    Cellulose from plant biomass can serve as a sustainable feedstock for fuels, chemicals and polymers that are currently produced from petroleum. In order to enhance economic feasibility, the efficiency of cell wall deconstruction needs to be enhanced. With the use of genetic and biotechnological approaches cell wall composition can be modified in such a way that interactions between the major cell wall polymers—cellulose, hemicellulosic polysaccharides and lignin—are altered. Some of the resulting plants are compromised in their growth and development, but this may be caused in part by the plant's overcompensation for metabolic perturbances. In other cases novel structures have been introduced in the cell wall without negative effects. The first field studies with engineered bioenergy crops look promising, while detailed structural analyses of cellulose synthase offer new opportunities to modify cellulose itself.

  5. Utilizing Core–Shell Fibrous Collagen-Alginate Hydrogel Cell Delivery System for Bone Tissue Engineering

    PubMed Central

    Perez, Roman A.; Kim, Meeju; Kim, Tae-Hyun; Kim, Joong-Hyun; Lee, Jae Ho; Park, Jeong-Hui; Knowles, Jonathan C.

    2014-01-01

    Three-dimensional matrices that encapsulate and deliver stem cells with defect-tuned formulations are promising for bone tissue engineering. In this study, we designed a novel stem cell delivery system composed of collagen and alginate as the core and shell, respectively. Mesenchymal stem cells (MSCs) were loaded into the collagen solution and then deposited directly into a fibrous structure while simultaneously sheathing with alginate using a newly designed core–shell nozzle. Alginate encapsulation was achieved by the crosslinking within an adjusted calcium-containing solution that effectively preserved the continuous fibrous structure of the inner cell-collagen part. The constructed hydrogel carriers showed a continuous fiber with a diameter of ∼700–1000 μm for the core and 200–500 μm for the shell area, which was largely dependent on the alginate concentration (2%–5%) as well as the injection rate (20–80 mL/h). The water uptake capacity of the core–shell carriers was as high as 98%, which could act as a pore channel to supply nutrients and oxygen to the cells. Degradation of the scaffolds showed a weight loss of ∼22% at 7 days and ∼43% at 14 days, suggesting a possible role as a degradable tissue-engineered construct. The MSCs encapsulated within the collagen core showed excellent viability, exhibiting significant cellular proliferation up to 21 days with levels comparable to those observed in the pure collagen gel matrix used as a control. A live/dead cell assay also confirmed similar percentages of live cells within the core–shell carrier compared to those in the pure collagen gel, suggesting the carrier was cell compatible and was effective for maintaining a cell population. Cells allowed to differentiate under osteogenic conditions expressed high levels of bone-related genes, including osteocalcin, bone sialoprotein, and osteopontin. Further, when the core–shell fibrous carriers were implanted in a rat calvarium defect, the bone

  6. Controllable delivery of small-molecule compounds to targeted cells utilizing carbon nanotubes.

    PubMed

    Su, Zhengding; Zhu, Shuihan; Donkor, Apraku D; Tzoganakis, Costas; Honek, John F

    2011-05-11

    Carbon nanotubes (CNTs) have emerged as a new alternative and efficient tool for transporting molecules with biotechnological and biomedical applications, because of their remarkable physicochemical properties. Encapsulation of functional molecules into the hollow chambers of CNTs can not only stabilize encapsulated molecules but also generate new nanodevices. In this work, we have demonstrated that CNTs can function as controllable carriers to transport small-molecule compounds (SMCs) loaded inside their hollow tunnels onto targeted cells. Using indole as model compound, CNTs can protect indole molecules during transportation. Labeling indole-loaded CNTs (indole@CNTs) with EphB4-binding peptides generates cell-homing indole@CNTs (CIDs). CIDs can selectively target EphB4-expressing cells and release indole onto cell surfaces by near-infrared (NIR) irradiation. Released indole molecules exhibit significant cell-killing effects without causing local overheating. This establishes CNTs as excellent near-infrared controllable delivery vehicles for SMCs as selective cell-killing agents.

  7. Conceptual designs for utility load-leveling battery with Li/FeS cells

    SciTech Connect

    Zivi, S. M.; Kacinskas, H.; Pollack, I.; Chilenskas, A. A.; Grieve, W.; McFarland, B. L.; Sudar, S.

    1980-07-01

    In 1978, a conceptual design of a 100 MW-h load-leveling battery system having Li alloy/FeS cells was developed as a result of a joint effort between ANL and Rockwell International. In this conceptual design, the submodule, which was the basic replaceable unit for the system, had a capacity of 240 kW-h and consisted of ninety-six 2.5 kW-h cells. However, a study by Rockwell indicated that the cost for battery hardware, $60 to 80/kW-h (cells and converters not included), was too high. Most of this cost was contributed by the submodule structure and the charge equalization scheme, which was the same as that developed for electric-vehicle batteries. In 1979, subsequent efforts were concentrated on lowering these hardware costs and resulted in the development of three modified designs, which are presented in this report. The first, developed at ANL, consisted of a 30 kW-h cell/submodule and the electric-vehicle equalization scheme. The hardware cost for this modified design was quite low, about $25/kW-h; however, this design was eventually rejected owing to the apparent impracticality of such a large cell. The two other modified designs had more conservative cell designs. One of them, developed at ANL, consisted of a 120 kW-h submodule consisting of one hundred 1.2 kW-h cells; the other, developed at Rockwell, consisted of a 1020 kW-h submodule consisting of four hundred and eight 2.5 kW-h cells. For both of these designs, an alternative method of equalization, in which fixed resistance shunts are used on each cell, was proposed; this equalization method adds little equipment cost to the system and only sacrifices about 4% of the coulombic and energy efficiencies. The cost of battery hardware for these two designs was estimated to be acceptable, about $22 to 60/kW-h. Some questions remain on the assumed capabilities of the cells and the feasibility of the battery hardware.

  8. Enhanced Antiproliferative Effect of Carboplatin in Cervical Cancer Cells Utilizing Folate-Grafted Polymeric Nanoparticles

    NASA Astrophysics Data System (ADS)

    Ji, Jing; Zuo, Ping; Wang, Yue-Ling

    2015-11-01

    Carboplatin (CRB) possesses superior anticancer effect in cervical cancer cells with lower incidence of side effects compared to that of cisplatin. However, CRB suffers from severe side effects due to undesirable tissue distributions which contribute to the low therapeutic efficacy. Here, we report a unique folic acid-conjugated chitosan-coated poly( d- l-lactideco-glycolide) (PLGA) nanoparticles (FPCC) prepared for the selective delivery of carboplatin to the cervical cancer cells. The particles were nanosized and spherical shaped with size less than <200 nm. The presence of protective chitosan layer controlled the overall release rate of CRB from chitosan-coated PLGA nanoparticles (PCC) and FPCC. FPCC displayed a higher cellular uptake capacity in HeLa cells than compared to non-targeted nanoparticles. Selective uptake of FPCC was due to an interaction of folic acid (FA) with the folate receptors alpha (FRs-α) which is overexpressed on the HeLa and promoted active targeting. These results indicated that FPCC had a specific affinity for the cancerous, HeLa cells owing to ligand-receptor (FA-FR-α) recognition. Consistently, FPCC showed superior cytotoxic effect than any other formulations. The IC50 (concentration of the drug required to kill 50 % of the cells) value of FPCC was 0.65 μg/ml while it was 1.08, 1.56, and 2.35 μg/ml for PCC, PLGA NP, and free CRB, respectively. Consistent with the cytotoxicity assay, FPCC induced higher fraction of early as well as late apoptosis cells. Especially, FPCC induced nearly 45 % of early apoptosis cells and more than 35 % in late apoptosis. Therefore, we propose that folate-conjugated nanoparticles might have potential applications in cervical cancer therapy.

  9. Involvement of microtubules in lipoprotein degradation and utilization for steroidogenesis in cultured rat luteal cells

    SciTech Connect

    Rajan, V.P.; Menon, K.M.

    1985-12-01

    Cells isolated from superovulated rat ovaries metabolize low density lipoprotein (LDL) and high density lipoprotein (HDL) of human or rat origin and use the lipoprotein-derived cholesterol as a precursor for progesterone production. Under in vitro conditions, both lipoproteins are internalized and degraded in the lysosomes, although degradation of HDL is of lower magnitude than that of LDL. In this report we have examined the role of cellular microtubules in the internalization and degradation of human LDL and HDL in cultured rat luteal cells. The microtubule depolymerizing agents colchicine, podophyllotoxin, vinblastine, and nocodazole as well as taxol, deuterium oxide, and dimethyl sulfoxide, which are known to rapidly polymerize cellular tubulin into microtubules, were used to block the function of microtubules. When these antimicrotubule agents were included in the incubations, degradation of the apolipoproteins of (/sup 125/I)iodo-LDL and (/sup 125/I)iodo-HDL by the luteal cells was inhibited by 50-85% compared to untreated control values. Maximum inhibitory effects were observed when the cells were preincubated with the inhibitor for at least 4 h at 37 C before treatment with the labeled lipoprotein. Lipoprotein-stimulated progesterone production by luteal cells was also inhibited by 50% or more in the presence of antimicrotubule agents. However, basal and hCG-stimulated progesterone production were unaffected by these inhibitors. The binding of (/sup 125/I)iodo-LDL and (/sup 125/I)iodo-HDL to luteal cell plasma membrane receptors was not affected by the microtubule inhibitors. Although binding was unaffected and degradation was impaired in the presence of the inhibitors, there was no detectable accumulation of undegraded lipoprotein within the cells during the 24 h of study.

  10. Efficient preparation of graphene liquid cell utilizing direct transfer with large-area well-stitched graphene

    NASA Astrophysics Data System (ADS)

    Sasaki, Yuki; Kitaura, Ryo; Yuk, Jong Min; Zettl, Alex; Shinohara, Hisanori

    2016-04-01

    By utilizing graphene-sandwiched structures recently developed in this laboratory, we are able to visualize small droplets of liquids in nanometer scale. We have found that small water droplets as small as several tens of nanometers sandwiched by two single-layer graphene are frequently observed by TEM. Due to the electron beam irradiation during the TEM observation, these sandwiched droplets are frequently moving from one place to another and are subjected to create small bubbles inside. The synthesis of a large area single-domain graphene of high-quality is essential to prepare the graphene sandwiched cell which safely encapsulates the droplets in nanometer size.

  11. Investigation of the effects of external current systems on the MAGSAT data utilizing grid cell modeling techniques

    NASA Technical Reports Server (NTRS)

    Klumpar, D. M. (Principal Investigator)

    1982-01-01

    The feasibility of modeling magnetic fields due to certain electrical currents flowing in the Earth's ionosphere and magnetosphere was investigated. A method was devised to carry out forward modeling of the magnetic perturbations that arise from space currents. The procedure utilizes a linear current element representation of the distributed electrical currents. The finite thickness elements are combined into loops which are in turn combined into cells having their base in the ionosphere. In addition to the extensive field modeling, additional software was developed for the reduction and analysis of the MAGSAT data in terms of the external current effects. Direct comparisons between the models and the MAGSAT data are possible.

  12. Androgen and FSH synergistically stimulate lipoprotein degradation and utilization by ovary granulosa cells

    SciTech Connect

    Schreiber, J.R.; Nakamura, K.; Schmit, V.; Weinstein, D.B.

    1984-01-01

    Androgen can directly modulate the induction of steroidogenic enzymes by FSH (follicle stimulating hormone) in ovary granulosa cells. In studies of its mechanism of action, the authors examined the androgen effect on granulosa cell interaction with lipoproteins, the physiologic source of cholesterol. After granulosa cells were cultured for 48 hours with and without androgen and/or FSH, the cells were incubated for 24 hours with /sup 125/I-lipoproteins (human high density lipoprotein (HDL), rat HDL, or human low density lipoprotein (LDL)). The media were then analyzed for lipoprotein protein coat degradation products (mainly /sup 125/I-monoiodotyrosine) and progestin (mainly 20 alpha-dihydroprogesterone (20 alpha-DHP)). In the absence of FSH and androgen, 2 X 10(5) granulosa cells degraded basal levels of all three lipoproteins, but produced no measurable 20 alpha-DHP. The addition of 10(-7) M androstenedione (A), testosterone (T), or 5 alpha-dihydrotestosterone (DHT) had no effect on lipoprotein protein degradation or 20 alpha-DHP production. FSH alone stimulated lipoprotein protein degradation by 50 to 300% while the addition of androgen synergistically augmented the FSH-stimulated 20 alpha-DHP production as well as protein coat degradation of all three lipoproteins. DHT and T were both effective, indicating that androgens themselves, and not estrogen products, were responsible for the effect on lipoprotein protein degradation and 20 alpha-DHP production.

  13. Utilization of Putrescine in Tobacco Cell Lines Resistant to Inhibitors of Polyamine Synthesis 1

    PubMed Central

    Hiatt, Andrew; Malmberg, Russell L.

    1988-01-01

    Three tobacco cell lines have been analyzed which are resistant to lethal inhibitors of either putrescine production or conversion of putrescine into polyamines. Free and conjugated putrescine pools, the enzymic activities (arginine, ornithine, and S-adenosylmethionine decarboxylases), and the growth characteristics during acidic stress were measured in suspension cultures of each cell line. One cell line, resistant to difluoromethylornithine (Dfr1) had a very low level of ornithine decarboxylase activity which was half insensitive to the inhibitor in vitro. Intracellular free putrescine in Dfr1 was elevated 10-fold which was apparently due to a 20-fold increase in the arginine decarboxylase activity. The increased free putrescine titer was not reflected in an increased level of spermidine, spermine, or putrescine conjugation. Dfr1 cultures survived acidic stress at molarities which were lethal to wild type cultures. Two other mutants, resistant to methylglyoxal bis(guanylhydrazone) (Mgr3, Mgr12), had near normal levels of the three decarboxylases and normal titers of free putrescine, spermidine, and spermine. Both mutants however had elevated levels of conjugated putrescine. Mgr12 had an increased sensitivity to acidic medium. These results suggest that increased levels of free putrescine production may enhance the ability of tobacco cells to survive acid stress. This was supported by the observation that cytotoxic effects of inhibiting arginine decarboxylase in wild type cell lines were dependent on the acidity of the medium. Images Fig. 1 PMID:16665927

  14. Evaluation of Gas-Cooled Pressurized Phosphoric Acid Fuel Cells for Electric Utility Power Generation

    NASA Technical Reports Server (NTRS)

    Faroque, M.

    1983-01-01

    Gas cooling is a more reliable, less expensive and a more simple alternative to conventional liquid cooling for heat removal from the phosphoric acid fuel cell (PAFC). The feasibility of gas-cooling was already demonstrated in atmospheric pressure stacks. Theoretical and experimental investigations of gas-cooling for pressurized PAFC are presented. Two approaches to gas cooling, Distributed Gas-Cooling (DIGAS) and Separated Gas-Cooling (SGC) were considered, and a theoretical comparison on the basis of cell performance indicated SGC to be superior to DIGAS. The feasibility of SGC was experimentally demonstrated by operating a 45-cell stack for 700 hours at pressure, and determining thermal response and the effect of other related parameters.

  15. Electrochemical electron paramagnetic resonance utilizing loop gap resonators and micro-electrochemical cells.

    PubMed

    Tamski, Mika A; Macpherson, Julie V; Unwin, Patrick R; Newton, Mark E

    2015-09-28

    A miniaturised electrochemical cell design for Electron Paramagnetic Resonance (EPR) studies is reported. The cell incorporates a Loop Gap Resonator (LGR) for EPR investigation of electrochemically generated radicals in aqueous (and other large dielectric loss) samples and achieves accurate potential control for electrochemistry by using micro-wires as working electrodes. The electrochemical behaviour of the cell is analysed with COMSOL finite element models and the EPR sensitivity compared to a commercial TE011 cavity resonator using 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL) as a reference. The electrochemical EPR performance is demonstrated using the reduction of methyl viologen as a redox probe in both water and acetonitrile. The data reported herein suggest that sub-micromolar concentrations of radical species can be detected in aqueous samples with accurate potential control, and that subtle solution processes coupled to electron transfer, such as comproportionation reactions, can be studied quantitatively using EPR. PMID:26291423

  16. Electrochemical electron paramagnetic resonance utilizing loop gap resonators and micro-electrochemical cells.

    PubMed

    Tamski, Mika A; Macpherson, Julie V; Unwin, Patrick R; Newton, Mark E

    2015-09-28

    A miniaturised electrochemical cell design for Electron Paramagnetic Resonance (EPR) studies is reported. The cell incorporates a Loop Gap Resonator (LGR) for EPR investigation of electrochemically generated radicals in aqueous (and other large dielectric loss) samples and achieves accurate potential control for electrochemistry by using micro-wires as working electrodes. The electrochemical behaviour of the cell is analysed with COMSOL finite element models and the EPR sensitivity compared to a commercial TE011 cavity resonator using 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL) as a reference. The electrochemical EPR performance is demonstrated using the reduction of methyl viologen as a redox probe in both water and acetonitrile. The data reported herein suggest that sub-micromolar concentrations of radical species can be detected in aqueous samples with accurate potential control, and that subtle solution processes coupled to electron transfer, such as comproportionation reactions, can be studied quantitatively using EPR.

  17. Three-Dimensional Spheroid Cell Culture Model for Target Identification Utilizing High-Throughput RNAi Screens.

    PubMed

    Iles, LaKesla R; Bartholomeusz, Geoffrey A

    2016-01-01

    The intrinsic limitations of 2D monolayer cell culture models have prompted the development of 3D cell culture model systems for in vitro studies. Multicellular tumor spheroid (MCTS) models closely simulate the pathophysiological milieu of solid tumors and are providing new insights into tumor biology as well as differentiation, tissue organization, and homeostasis. They are straightforward to apply in high-throughput screens and there is a great need for the development of reliable and robust 3D spheroid-based assays for high-throughput RNAi screening for target identification and cell signaling studies highlighting their potential in cancer research and treatment. In this chapter we describe a stringent standard operating procedure for the use of MCTS for high-throughput RNAi screens. PMID:27581289

  18. Utility of tricalcium phosphate and osteogenic matrix cell sheet constructs for bone defect reconstruction

    PubMed Central

    Ueha, Tomoyuki; Akahane, Manabu; Shimizu, Takamasa; Uchihara, Yoshinobu; Morita, Yusuke; Nitta, Naoya; Kido, Akira; Inagaki, Yusuke; Kawate, Kenji; Tanaka, Yasuhito

    2015-01-01

    AIM: To determine the effects of transplanting osteogenic matrix cell sheets and beta-tricalcium phosphate (TCP) constructs on bone formation in bone defects. METHODS: Osteogenic matrix cell sheets were prepared from bone marrow stromal cells (BMSCs), and a porous TCP ceramic was used as a scaffold. Three experimental groups were prepared, comprised of TCP scaffolds (1) seeded with BMSCs; (2) wrapped with osteogenic matrix cell sheets; or (3) both. Constructs were implanted into a femoral defect model in rats and bone growth was evaluated by radiography, histology, biochemistry, and mechanical testing after 8 wk. RESULTS: In bone defects, constructs implanted with cell sheets showed callus formation with segmental or continuous bone formation at 8 wk, in contrast to TCP seeded with BMSCs, which resulted in bone non-union. Wrapping TCP constructs with osteogenic matrix cell sheets increased their osteogenic potential and resulting bone formation, compared with conventional bone tissue engineering TCP scaffolds seeded with BMSCs. The compressive stiffness (mean ± SD) values were 225.0 ± 95.7, 30.0 ± 11.5, and 26.3 ± 10.6 MPa for BMSC/TCP/Sheet constructs with continuous bone formation, BMSC/TCP/Sheet constructs with segmental bone formation, and BMSC/TCP constructs, respectively. The compressive stiffness of BMSC/TCP/Sheet constructs with continuous bone formation was significantly higher than those with segmental bone formation and BMSC/TCP constructs. CONCLUSION: This technique is an improvement over current methods, such as TCP substitution, and is useful for hard tissue reconstruction and inducing earlier bone union in defects. PMID:26131318

  19. Human Coronavirus NL63 Utilizes Heparan Sulfate Proteoglycans for Attachment to Target Cells

    PubMed Central

    Milewska, Aleksandra; Zarebski, Miroslaw; Nowak, Paulina; Stozek, Karol; Potempa, Jan

    2014-01-01

    ABSTRACT Human coronavirus NL63 (HCoV-NL63) is an alphacoronavirus that was first identified in 2004 in the nasopharyngeal aspirate from a 7-month-old patient with a respiratory tract infection. Previous studies showed that HCoV-NL63 and the genetically distant severe acute respiratory syndrome (SARS)-CoV employ the same receptor for host cell entry, angiotensin-converting enzyme 2 (ACE2), but it is largely unclear whether ACE2 interactions are sufficient to allow HCoV-NL63 binding to cells. The present study showed that directed expression of angiotensin-converting enzyme 2 (ACE2) on cells previously resistant to HCoV-NL63 renders them susceptible, showing that ACE2 protein acts as a functional receptor and that its expression is required for infection. However, comparative analysis showed that directed expression or selective scission of the ACE2 protein had no measurable effect on virus adhesion. In contrast, binding of HCoV-NL63 to heparan sulfates was required for viral attachment and infection of target cells, showing that these molecules serve as attachment receptors for HCoV-NL63. IMPORTANCE ACE2 protein was proposed as a receptor for HCoV-NL63 already in 2005, but an in-depth analysis of early events during virus infection had not been performed thus far. Here, we show that the ACE2 protein is required for viral entry but that it is not the primary binding site on the cell surface. Conducted research showed that heparan sulfate proteoglycans function as adhesion molecules, increasing the virus density on cell surface and possibly facilitating the interaction between HCoV-NL63 and its receptor. Obtained results show that the initial events during HCoV-NL63 infection are more complex than anticipated and that a newly described interaction may be essential for understanding the infection process and, possibly, also assist in drug design. PMID:25187545

  20. Nano-photonic organic solar cell architecture for advanced light management utilizing dual photonic crystals

    NASA Astrophysics Data System (ADS)

    Peer, Akshit; Biswas, Rana

    2015-09-01

    Organic solar cells have rapidly increasing efficiencies, but typically absorb less than half of the incident solar spectrum. To increase broadband light absorption, we rigorously design experimentally realizable solar cell architectures based on dual photonic crystals. Our optimized architecture consists of a polymer microlens at the air-glass interface, coupled with a photonic-plasmonic crystal at the metal cathode. The microlens focuses light on the periodic nanostructure that generates strong light diffraction. Waveguiding modes and surface plasmon modes together enhance long wavelength absorption in P3HT-PCBM. The architecture has a period of 500 nm, with absorption and photocurrent enhancement of 49% and 58%, respectively.

  1. Utilization of Microgravity Bioreactor for Differentiation and Growth of Human Vascular Endothelial Cells

    NASA Technical Reports Server (NTRS)

    Chen, Chu-Huang; Pellis, Neal R.

    1997-01-01

    The goal was to delineate mechanisms of genetic responses to angiogenic stimulation of human coronary arterial and dermal microvascular endothelial cells during exposure to microgravity. The NASA-designed rotating-wall vessel was used to create a three-dimensional culture environment with low shear-stress and microgravity simulating that in space. The primary specific aim was to determine whether simulated microgravity enhances endothelial cell growth and whether the growth enhancement is associated by augmented expression of Basic Fibroblast Growth Factor (BFGF) and c-fos, an immediate early gene and component of the transcription factor AP-1.

  2. PANTOTHENIC ACID AND THE UTILIZATION OF GLUCOSE BY LIVING AND CELL-FREE SYSTEMS.

    PubMed

    Teague, P C; Williams, R J

    1942-07-20

    1. Added pantothenic acid was found to have no appreciable effect on the fermentation of glucose when used in conjunction with preparations of dialyzed yeast maceration juice or acetone-precipitated yeast maceration juice. 2. Addition of pantothenic acid failed to affect the rate of phosphorylation of glucose or the rate of decarboxylation of pyruvic acid by yeast maceration juice. 3. Pantothenic acid showed no effect on the rate of glycolysis by homogenized deficient chick tissues. 4. The accelerating effect of pantothenic acid on fermentation by deficient yeast cells was found to be accompanied by a "binding" of pantothenic acid by the yeast cells.

  3. Fabrication and characterization of protonic-ceramic fuel cells and electrolysis cells utilizing infiltrated lanthanum nickelate electrodes

    NASA Astrophysics Data System (ADS)

    Babiniec, Sean M.

    High-temperature protonic ceramics (HTPCs) have gained interest as fuel cell and electrolysis cell electrolytes, as well as hydrogen separation membranes. The transport of hydrogen as opposed to oxygen results in several benefits and applications, including higher fuel efficiency, dehydrogenation of fuel streams, and hydrogen-based chemical synthesis. However, limited work has been done in the development of air/steam electrodes for these devices. This work presents the characterization of lanthanum nickelate, La 2NiO4+delta (LN), as a potential air/steam electrode material for use with BaCe0.2Zr0.7Y0.1O3-delta (BCZY27) HTPC electrolytes fabricated by the solid-state reactive sintering technique. Two types of devices were made; a symmetric cell used for electrode characterization, and a full fuel cell/electrolysis cell used for device performance characterization. The symmetric cell consists of a 1 mm thick BCZY27 substrate with identical air/steam electrodes on both sides. Air/steam electrodes were made by infiltrating ˜ 50 nm lanthanum nickelate nanoparticles into a BCZY27 porous backbone. The fuel cell/electrolysis cell consists of a 1mm thick Ni/BCZY27 anode support, a 25 mum thick BCZY27 electrolyte, and a 50 mum thick porous BCZY27 backbone infiltrated with lanthanum nickelate. Through symmetric cell testing, it was found that the electrode polarization resistance decreases with increasing oxygen content, indicating good oxygen reduction reaction characteristics. A minimum polarization resistance was found as 2.58 Ohm-cm2 in 3% humidied oxygen at 700 °C. Full cell testing revealed a peak power density of 27 mW-cm-2 at 700 °C. Hydrogen flux measurements were also taken in the both galvanic/post-galvanic and electrolytic operation. Galvanic/post-galvanic fluxes exhibit a very high faradaic efficiency. However, electrolytic hydrogen fluxes were much lower than the calculated hydrogen faradaic flux, indicating a different charge carrier other than protons is

  4. [Effect of 5-FU on the utilization of purine and pyrimidine by human gastric cancer cells (KATO III)].

    PubMed

    Usami, M; Wang, J; Yasuda, I; Saitoh, Y; Yumisashi, T; Abe, K

    1995-05-01

    Effect of utilization of purine and pyrimidine in the culture medium by human gastric cancer cells (KATO III) was evaluated. Nucleosides mixture solution (OG-VI), consisting of inosine, guanosine 5' monophosphate (5'GMP), cytidine, uridine and thymidine (4: 4: 4: 3: 1 in molar ratio) was used and their levels in the culture medium was measured by HPLC after 3 day culture. Purine, inosine and 5' GMP, in the medium almost decreased and purine base, xanthine and hypoxanthine levels increased, but changes in pyrimidine level were minimal. 5-FU decreased purine and increased pyrimidine consumption. Addition of nucleosides mixture did not enhance the cellular proliferation, but inhibited growth when given in higher concentrations. Nucleoside mixture solution enhanced growth inhibition by 5-FU and it is a potential biochemical modulator of 5-FU metabolism in human cancer cells. PMID:7755382

  5. Expression and purification of 15 kDa granulysin utilizing an insect cell secretion system.

    PubMed

    Finn, Michael W; Clayberger, Carol; Krensky, Alan M

    2011-01-01

    Granulysin is an antimicrobial and proinflammatory protein expressed in activated human T cells and natural killer cells. A single mRNA produces the 15 kDa isoform which is then cleaved at the amino and carboxy termini to produce the 9 kDa isoform. Recombinant 9 kDa granulysin has been studied in detail but little is known about the function of the 15 kDa isoform, and no protocol has been published describing expression and purification of this form. Two commercially available preparations of the recombinant 15 kDa granulysin contain tags that may affect function. Here we describe for the first time a method to produce 15 kDa granulysin as a secreted protein from insect cells. The 15 kDa granulysin is purified using a HiTrap Heparin column and a Resource S column. A typical a yield of purified 15 kDa granulysin is 0.6 mg/L of insect cell supernatant.

  6. Utilization of Lymphoblastoid Cell Lines as a System for the Molecular Modeling of Autism

    ERIC Educational Resources Information Center

    Baron, Colin A.; Liu, Stephenie Y.; Hicks, Chindo; Gregg, Jeffrey P.

    2006-01-01

    In order to provide an alternative approach for understanding the biology and genetics of autism, we performed statistical analysis of gene expression profiles of lymphoblastoid cell lines derived from children with autism and their families. The goal was to assess the feasibility of using this model in identifying autism-associated genes.…

  7. Utilizing the green alga Chlamydomonas reinhardtii for microbial electricity generation: a living solar cell.

    PubMed

    Rosenbaum, Miriam; Schröder, Uwe; Scholz, Fritz

    2005-10-01

    By employing living cells of the green alga Chlamydomonas reinhardtii, we demonstrate the possibility of direct electricity generation from microbial photosynthetic activity. The presented concept is based on an in situ oxidative depletion of hydrogen, photosynthetically produced by C. reinhardtii under sulfur-deprived conditions, by polymer-coated electrocatalytic electrodes. PMID:15696280

  8. Utility of Dexrazoxane for the Attenuation of Epirubicin-Induced Genetic Alterations in Mouse Germ Cells

    PubMed Central

    Ahmad, Sheikh F.; Ansaria, Mushtaq A.; Nadeem, Ahmed; Al-Shabanah, Othman A.; Al-Harbi, Mohammed M.; Bakheet, Saleh A.

    2016-01-01

    Dexrazoxane has been approved to treat anthracycline-induced cardiomyopathy and extravasation. However, the effect of dexrazoxane on epirubicin-induced genetic alterations in germ cells has not yet been reported. Thus, the aim of this study was to determine whether dexrazoxane modulates epirubicin-induced genetic damage in the germ cells of male mice. Our results show that dexrazoxane was not genotoxic at the tested doses. Furthermore, it protected mouse germ cells against epirubicin-induced genetic alterations as detected by the reduction in disomic and diploid sperm, spermatogonial chromosomal aberrations, and abnormal sperm heads. The attenuating effect of dexrazoxane was greater at higher dose, indicating a dose-dependent effect. Moreover, sperm motility and count were ameliorated by dexrazoxane pretreatment. Epirubicin induced marked biochemical changes characteristic of oxidative DNA damage including elevated 8-hydroxy-2ʹ-deoxyguanosine levels and reduction in reduced glutathione. Pretreatment of mice with dexrazoxane before epirubicin challenge restored these altered endpoints. We conclude that dexrazoxane may efficiently mitigate the epirubicin insult in male germ cells, and prevent the enhanced risk of abnormal reproductive outcomes and associated health risks. Thus, pretreating patients with dexrazoxane prior to epirubicin may efficiently preserve not only sperm quality but also prevent the transmission of genetic damage to future generations. PMID:27690233

  9. Utility of magnetic cell separation as a molecular sperm preparation technique.

    PubMed

    Said, Tamer M; Agarwal, Ashok; Zborowski, Maciej; Grunewald, Sonja; Glander, Hans-Juergen; Paasch, Uwe

    2008-01-01

    Assisted reproductive techniques (ARTs) have become the treatment of choice in many cases of infertility; however, the current success rates of these procedures remain suboptimal. Programmed cell death (apoptosis) most likely contributes to failed ART and to the decrease in sperm quality after cryopreservation. There is a likelihood that some sperm selected for ART will display features of apoptosis despite their normal appearance, which may be partially responsible for the low fertilization and implantation rates seen with ART. One of the features of apoptosis is the externalization of phosphatidylserine (PS) residues, which are normally present on the inner leaflet of the sperm plasma membrane. Colloidal superparamagnetic microbeads ( approximately 50 nm in diameter) conjugated with annexin V bind to PS and are used to separate dead and apoptotic spermatozoa by magnetic-activated cell sorting (MACS). Cells with externalized PS will bind to these microbeads, whereas nonapoptotic cells with intact membranes do not bind and could be used during ARTs. We have conducted a series of experiments to investigate whether the MACS technology could be used to improve ART outcomes. Our results clearly indicate that integrating MACS as a part of sperm preparation techniques will improve semen quality and cryosurvival rates by eliminating apoptotic sperm. Nonapoptotic spermatozoa prepared by MACS display higher quality in terms of routine sperm parameters and apoptosis markers. The higher sperm quality is represented by an increased oocyte penetration potential and cryosurvival rates. Thus, the selection of nonapoptotic spermatozoa by MACS should be considered to enhance ART success rates. PMID:18077822

  10. Dendritic Cell Based Vaccines that Utilize Myeloid Rather than Plasmacytoid Cells Offer a Superior Survival Advantage in Malignant Glioma

    PubMed Central

    Dey, Mahua; Chang, Alan L.; Miska, Jason; Wainwright, Derek A.; Ahmed, Atique U.; Balyasnikova, Irina V.; Pytel, Peter; Han, Yu; Tobias, Alex; Zhang, Lingjiao; Qiao, Jian; Lesniak, Maciej S.

    2015-01-01

    Dendritic cells (DC) are professional antigen presenting cells (APC) that are traditionally divided into two distinct subsets: myeloid DC (mDCs) and plasmacytoid DC (pDCs). pDCs are known for their ability to secrete large amount of IFN-α. Apart from IFN-α production, pDCs can also process antigen and induce T-cell immunity or tolerance. In several solid tumors, pDCs have been shown to play a critical role in promoting tumor immunosuppression. We investigated the role of pDCs in the process of glioma progression in the syngeneic murine model of glioma. We show that glioma-infiltrating pDCs are the major APC in glioma and are deficient in IFN-α secretion (p < 0.05). pDC depletion leads to increased survival of the mice bearing intracranial tumor by decreasing the number of regulatory T-cells (Treg) and by decreasing the suppressive capabilities of Tregs. We subsequently compared the ability of mDCs and pDCs to generate effective anti-glioma immunity in a GL261-OVA mouse model of glioma. Our data suggest that mature pDCs and mDCs isolated from naïve mice can be effectively activated and loaded with SIINFEKL antigen in vitro. Upon intra-dermal injection in the hind leg, a fraction of both types of DCs migrate to the brain and lymph nodes.. Compared to mice vaccinated with pDC or control mice, mice vaccinated with mDCs generated a robust Th1 type immune response, characterized by high frequency of CD4+Tbet+ T-cells and CD8+Siinfekel+ T-cells. This robust anti-tumor T-cell response resulted in tumor eradication and long-term survival in 60% of the animals (p<0.001). PMID:26026061

  11. Essential Domains of Anaplasma phagocytophilum Invasins Utilized to Infect Mammalian Host Cells

    PubMed Central

    Seidman, David; Hebert, Kathryn S.; Truchan, Hilary K.; Miller, Daniel P.; Tegels, Brittney K.; Marconi, Richard T.; Carlyon, Jason A.

    2015-01-01

    Anaplasma phagocytophilum causes granulocytic anaplasmosis, an emerging disease of humans and domestic animals. The obligate intracellular bacterium uses its invasins OmpA, Asp14, and AipA to infect myeloid and non-phagocytic cells. Identifying the domains of these proteins that mediate binding and entry, and determining the molecular basis of their interactions with host cell receptors would significantly advance understanding of A. phagocytophilum infection. Here, we identified the OmpA binding domain as residues 59 to 74. Polyclonal antibody generated against a peptide spanning OmpA residues 59 to 74 inhibited A. phagocytophilum infection of host cells and binding to its receptor, sialyl Lewis x (sLex-capped P-selectin glycoprotein ligand 1. Molecular docking analyses predicted that OmpA residues G61 and K64 interact with the two sLex sugars that are important for infection, α2,3-sialic acid and α1,3-fucose. Amino acid substitution analyses demonstrated that K64 was necessary, and G61 was contributory, for recombinant OmpA to bind to host cells and competitively inhibit A. phagocytophilum infection. Adherence of OmpA to RF/6A endothelial cells, which express little to no sLex but express the structurally similar glycan, 6-sulfo-sLex, required α2,3-sialic acid and α1,3-fucose and was antagonized by 6-sulfo-sLex antibody. Binding and uptake of OmpA-coated latex beads by myeloid cells was sensitive to sialidase, fucosidase, and sLex antibody. The Asp14 binding domain was also defined, as antibody specific for residues 113 to 124 inhibited infection. Because OmpA, Asp14, and AipA each contribute to the infection process, it was rationalized that the most effective blocking approach would target all three. An antibody cocktail targeting the OmpA, Asp14, and AipA binding domains neutralized A. phagocytophilum binding and infection of host cells. This study dissects OmpA-receptor interactions and demonstrates the effectiveness of binding domain-specific antibodies

  12. Hydrogen Fuel Cells and Storage Technology: Fundamental Research for Optimization of Hydrogen Storage and Utilization

    SciTech Connect

    Perret, Bob; Heske, Clemens; Nadavalath, Balakrishnan; Cornelius, Andrew; Hatchett, David; Bae, Chusung; Pang, Tao; Kim, Eunja; Hemmers, Oliver

    2011-03-28

    Design and development of improved low-cost hydrogen fuel cell catalytic materials and high-capacity hydrogenn storage media are paramount to enabling the hydrogen economy. Presently, effective and durable catalysts are mostly precious metals in pure or alloyed form and their high cost inhibits fuel cell applications. Similarly, materials that meet on-board hydrogen storage targets within total mass and volumetric constraints are yet to be found. Both hydrogen storage performance and cost-effective fuel cell designs are intimately linked to the electronic structure, morphology and cost of the chosen materials. The FCAST Project combined theoretical and experimental studies of electronic structure, chemical bonding, and hydrogen adsorption/desorption characteristics of a number of different nanomaterials and metal clusters to develop better fundamental understanding of hydrogen storage in solid state matrices. Additional experimental studies quantified the hydrogen storage properties of synthesized polyaniline(PANI)/Pd composites. Such conducting polymers are especially interesting because of their high intrinsic electron density and the ability to dope the materials with protons, anions, and metal species. Earlier work produced contradictory results: one study reported 7% to 8% hydrogen uptake while a second study reported zero hydrogen uptake. Cost and durability of fuel cell systems are crucial factors in their affordability. Limits on operating temperature, loss of catalytic reactivity and degradation of proton exchange membranes are factors that affect system durability and contribute to operational costs. More cost effective fuel cell components were sought through studies of the physical and chemical nature of catalyst performance, characterization of oxidation and reduction processes on system surfaces. Additional development effort resulted in a new hydrocarbon-based high-performance sulfonated proton exchange membrane (PEM) that can be manufactured at low

  13. Freezing without Ice Crystal Damage: Semithin and Ultrathin Frozen Sections of Ethanol-Infiltrated Tissue for Microscopy, with Applications to Immunocytochemistry

    NASA Astrophysics Data System (ADS)

    Christensen, A. Kent; Lowry, Terry B.

    1995-10-01

    Ethanol (ethyl alcohol) has long been a standard reagent used in preparing tissues for light and electron microscopy. After fixation, tissues are usually dehydrated with ethanol before being embedded in paraffin or plastic. In this study we show that the ethanol-infiltrated tissue can be frozen and sectioned directly without embedding. When tissue impregnated with ethanol is cooled below about [minus sign]117°C with liquid nitrogen, the ethanol solidifies without appreciable crystallization. The frozen tissue can then be sectioned in a commercial cryoultramicrotome that is set at [minus sign]155 to [minus sign]170°C to produce semithin frozen sections (0.5 to 3 [mu]m thick) for light microscopy or ultrathin frozen sections (50 to 100 nm thick) for electron microscopy. Sections are picked up and mounted on glass slides or EM grids by means that are in current use for ice ultrathin frozen sectioning. Because there is no apparent freezing damage, the morphology in these ethanol frozen sections of unembedded tissue appears generally quite good, often resembling that obtained by conventional EM techniques. Examples are provided that illustrate the use of this material for immunocytochemistry at the light and electron microscope levels.

  14. Group 2 innate lymphoid cells utilize the IRF4-IL-9 module to coordinate epithelial cell maintenance of lung homeostasis.

    PubMed

    Mohapatra, A; Van Dyken, S J; Schneider, C; Nussbaum, J C; Liang, H-E; Locksley, R M

    2016-01-01

    Group 2 innate lymphoid cells (ILC2s) have an important role in acute allergic lung inflammation. Given their distribution and function, lung ILC2s are hypothesized to coordinate epithelial responses to the external environment; however, how barrier surveillance is linked to ILC2 activation remains unclear. Here, we demonstrate that alveolar type II cells are the main source of interleukin (IL)-33 and thymic stromal lymphopoietin (TSLP) generated in response to chitin or migratory helminths. IL-33 and TSLP synergistically induce an interferon regulatory factor 4 (IRF4)-IL-9 program in ILC2s, and autocrine IL-9 promotes rapid IL-5 and IL-13 production required for optimal epithelial responses in the conducting airways. Thus, ILC2s link alveolar function to regulation of airway flow, revealing a key interaction between resident lymphoid and structural cells that might underlie similar organizational hierarchies in other organs.

  15. Utility of measuring serum or red blood cell folate in the era of folate fortification of flour.

    PubMed

    Gilfix, Brian M

    2014-05-01

    Folic acid is an essential nutrient involved in one-carbon metabolism. Insufficient folate can result in megaloblastic anemia and an increased risk of neural tube defects. In response to the latter, some governments have mandated the fortification of flour with folate. This had resulted in a documented rise in the serum and red blood cell folate levels in the population. This has impacted the potential utility of folate measurements to detect folate deficiency in the clinical context. Folate measurements, whether done in serum or red blood cells, are subject to analytical variation, especially the latter, which also affects the utility of such measurements. Examining the literature reveals that in clinical situations, generally <1% of the subjects will have folate deficiency regardless of potentially pre-disposing factors (e.g. anemia, anti-folate agents, inflammatory bowel disease). Data from our center for both pediatric and adult populations is presented that supports this observation. Consequently, there exists very few indications for folate determinations (unexplained macrocytosis, inborn errors of metabolism) and it may be more efficient to simply treat suspected cases. PMID:24486651

  16. Dendritic Cell-Based Vaccines that Utilize Myeloid Rather than Plasmacytoid Cells Offer a Superior Survival Advantage in Malignant Glioma.

    PubMed

    Dey, Mahua; Chang, Alan L; Miska, Jason; Wainwright, Derek A; Ahmed, Atique U; Balyasnikova, Irina V; Pytel, Peter; Han, Yu; Tobias, Alex; Zhang, Lingjiao; Qiao, Jian; Lesniak, Maciej S

    2015-07-01

    Dendritic cells (DCs) are professional APCs that are traditionally divided into two distinct subsets, myeloid DC (mDCs) and plasmacytoid DC (pDCs). pDCs are known for their ability to secrete large amounts of IFN-α. Apart from IFN-α production, pDCs can also process Ag and induce T cell immunity or tolerance. In several solid tumors, pDCs have been shown to play a critical role in promoting tumor immunosuppression. We investigated the role of pDCs in the process of glioma progression in the syngeneic murine model of glioma. We show that glioma-infiltrating pDCs are the major APC in glioma and are deficient in IFN-α secretion (p < 0.05). pDC depletion leads to increased survival of the mice bearing intracranial tumor by decreasing the number of regulatory T cells (Tregs) and by decreasing the suppressive capabilities of Tregs. We subsequently compared the ability of mDCs and pDCs to generate effective antiglioma immunity in a GL261-OVA mouse model of glioma. Our data suggest that mature pDCs and mDCs isolated from naive mice can be effectively activated and loaded with SIINFEKL Ag in vitro. Upon intradermal injection in the hindleg, a fraction of both types of DCs migrate to the brain and lymph nodes. Compared to mice vaccinated with pDC or control mice, mice vaccinated with mDCs generate a robust Th1 type immune response, characterized by high frequency of CD4(+)T-bet(+) T cells and CD8(+)SIINFEKEL(+) T cells. This robust antitumor T cell response results in tumor eradication and long-term survival in 60% of the animals (p < 0.001). PMID:26026061

  17. Dendritic Cell-Based Vaccines that Utilize Myeloid Rather than Plasmacytoid Cells Offer a Superior Survival Advantage in Malignant Glioma.

    PubMed

    Dey, Mahua; Chang, Alan L; Miska, Jason; Wainwright, Derek A; Ahmed, Atique U; Balyasnikova, Irina V; Pytel, Peter; Han, Yu; Tobias, Alex; Zhang, Lingjiao; Qiao, Jian; Lesniak, Maciej S

    2015-07-01

    Dendritic cells (DCs) are professional APCs that are traditionally divided into two distinct subsets, myeloid DC (mDCs) and plasmacytoid DC (pDCs). pDCs are known for their ability to secrete large amounts of IFN-α. Apart from IFN-α production, pDCs can also process Ag and induce T cell immunity or tolerance. In several solid tumors, pDCs have been shown to play a critical role in promoting tumor immunosuppression. We investigated the role of pDCs in the process of glioma progression in the syngeneic murine model of glioma. We show that glioma-infiltrating pDCs are the major APC in glioma and are deficient in IFN-α secretion (p < 0.05). pDC depletion leads to increased survival of the mice bearing intracranial tumor by decreasing the number of regulatory T cells (Tregs) and by decreasing the suppressive capabilities of Tregs. We subsequently compared the ability of mDCs and pDCs to generate effective antiglioma immunity in a GL261-OVA mouse model of glioma. Our data suggest that mature pDCs and mDCs isolated from naive mice can be effectively activated and loaded with SIINFEKL Ag in vitro. Upon intradermal injection in the hindleg, a fraction of both types of DCs migrate to the brain and lymph nodes. Compared to mice vaccinated with pDC or control mice, mice vaccinated with mDCs generate a robust Th1 type immune response, characterized by high frequency of CD4(+)T-bet(+) T cells and CD8(+)SIINFEKEL(+) T cells. This robust antitumor T cell response results in tumor eradication and long-term survival in 60% of the animals (p < 0.001).

  18. BdCESA7, BdCESA8, and BdPMT Utility Promoter Constructs for Targeted Expression to Secondary Cell-Wall-Forming Cells of Grasses

    PubMed Central

    Petrik, Deborah L.; Cass, Cynthia L.; Padmakshan, Dharshana; Foster, Cliff E.; Vogel, John P.; Karlen, Steven D.; Ralph, John; Sedbrook, John C.

    2016-01-01

    Utility vectors with promoters that confer desired spatial and temporal expression patterns are useful tools for studying gene and cellular function and for industrial applications. To target the expression of DNA sequences of interest to cells forming plant secondary cell walls, which generate most of the vegetative biomass, upstream regulatory sequences of the Brachypodium distachyon lignin biosynthetic gene BdPMT and the cellulose synthase genes BdCESA7 and BdCESA8 were isolated and cloned into binary vectors designed for Agrobacterium-mediated transformation of monocots. Expression patterns were assessed using the β-glucuronidase gene GUSPlus and X-glucuronide staining. All three promoters showed strong expression levels in stem tissue at the base of internodes where cell wall deposition is most active, in both vascular bundle xylem vessels and tracheids, and in interfascicular tissues, with expression less pronounced in developmentally older tissues. In leaves, BdCESA7 and BdCESA8 promoter-driven expression was strongest in leaf veins, leaf margins, and trichomes; relatively weaker and patchy expression was observed in the epidermis. BdPMT promoter-driven expression was similar to the BdCESA promoters expression patterns, including strong expression in trichomes. The intensity and extent of GUS staining varied considerably between transgenic lines, suggesting that positional effects influenced promoter activity. Introducing the BdPMT and BdCESA8 Open Reading Frames into BdPMT and BdCESA8 utility promoter binary vectors, respectively, and transforming those constructs into Brachypodium pmt and cesa8 loss-of-function mutants resulted in rescue of the corresponding mutant phenotypes. This work therefore validates the functionality of these utility promoter binary vectors for use in Brachypodium and likely other grass species. The identification, in Bdcesa8-1 T-DNA mutant stems, of an 80% reduction in crystalline cellulose levels confirms that the BdCESA8 gene is

  19. Solar cells utilizing pulsed-energy crystallized microcrystalline/polycrystalline silicon

    DOEpatents

    Kaschmitter, J.L.; Sigmon, T.W.

    1995-10-10

    A process for producing multi-terminal devices such as solar cells wherein a pulsed high energy source is used to melt and crystallize amorphous silicon deposited on a substrate which is intolerant to high processing temperatures, whereby the amorphous silicon is converted into a microcrystalline/polycrystalline phase. Dopant and hydrogenation can be added during the fabrication process which provides for fabrication of extremely planar, ultra shallow contacts which results in reduction of non-current collecting contact volume. The use of the pulsed energy beams results in the ability to fabricate high efficiency microcrystalline/polycrystalline solar cells on the so-called low-temperature, inexpensive plastic substrates which are intolerant to high processing temperatures.

  20. Low-cost, high-efficiency solar cells utilizing GaAs-on-Si technology

    SciTech Connect

    Vernon, S.M. )

    1993-04-01

    This report describes work to develop technology to deposit GaAs on Si using a nucleation layer of atomic-layer-epitaxy-grown GaAs or AlAs on Si. This ensures two-dimensional nucleation and should lead to fewer defects in the final GaAs layer. As an alternative, we also developed technology for depositing GaAs on sawtooth-patterned Si. Preliminary studies showed that this material can have a very low defect density, [approximately] 1 [times] 10[sup 5] cm[sup [minus]5], as opposed to our conventionally grown GaAs on SL which has a typical defect density of over 1 [times]10[sup 7] cm[sup [minus]2]. Using these two now methods of GaAs-on-Si material growth, we made solar cells that are expected to show higher efficiencies than those of previous cells.

  1. Solar cells utilizing pulsed-energy crystallized microcrystalline/polycrystalline silicon

    DOEpatents

    Kaschmitter, James L.; Sigmon, Thomas W.

    1995-01-01

    A process for producing multi-terminal devices such as solar cells wherein a pulsed high energy source is used to melt and crystallize amorphous silicon deposited on a substrate which is intolerant to high processing temperatures, whereby to amorphous silicon is converted into a microcrystalline/polycrystalline phase. Dopant and hydrogenization can be added during the fabrication process which provides for fabrication of extremely planar, ultra shallow contacts which results in reduction of non-current collecting contact volume. The use of the pulsed energy beams results in the ability to fabricate high efficiency microcrystalline/polycrystalline solar cells on the so-called low-temperature, inexpensive plastic substrates which are intolerant to high processing temperatures.

  2. Nanowires improved charge separation and light utilization in metal-oxide solar cells

    NASA Astrophysics Data System (ADS)

    Cheng, Wei-Yun; Lin, Yi-Feng; Lu, Shih-Yuan

    2011-08-01

    The power conversion efficiencies of electrodeposited Cu2O/ZnO p-n junction based solar cells are significantly improved by sandwiching a layer of spin-coated CdS nanowires (NWs) in between the electrochemically deposited Cu2O and ZnO layers. With the inclusion of the CdS NWs, there is observed a 5 fold improvement in the conversion efficiency, from 0.12% to 0.6%, as compared with that of the plain Cu2O/ZnO cell. The improvement is attributed to the enlarged p-n interface area and enhanced light harvesting, charge separation, and electron transport made possible by incorporating the single crystalline, relatively low band gap CdS NWs.

  3. Functional Genomics Screening Utilizing Mutant Mouse Embryonic Stem Cells Identifies Novel Radiation-Response Genes

    PubMed Central

    Loesch, Kimberly; Galaviz, Stacy; Hamoui, Zaher; Clanton, Ryan; Akabani, Gamal; Deveau, Michael; DeJesus, Michael; Ioerger, Thomas; Sacchettini, James C.; Wallis, Deeann

    2015-01-01

    Elucidating the genetic determinants of radiation response is crucial to optimizing and individualizing radiotherapy for cancer patients. In order to identify genes that are involved in enhanced sensitivity or resistance to radiation, a library of stable mutant murine embryonic stem cells (ESCs), each with a defined mutation, was screened for cell viability and gene expression in response to radiation exposure. We focused on a cancer-relevant subset of over 500 mutant ESC lines. We identified 13 genes; 7 genes that have been previously implicated in radiation response and 6 other genes that have never been implicated in radiation response. After screening, proteomic analysis showed enrichment for genes involved in cellular component disassembly (e.g. Dstn and Pex14) and regulation of growth (e.g. Adnp2, Epc1, and Ing4). Overall, the best targets with the highest potential for sensitizing cancer cells to radiation were Dstn and Map2k6, and the best targets for enhancing resistance to radiation were Iqgap and Vcan. Hence, we provide compelling evidence that screening mutant ESCs is a powerful approach to identify genes that alter radiation response. Ultimately, this knowledge can be used to define genetic variants or therapeutic targets that will enhance clinical therapy. PMID:25853515

  4. Utilization of fuel cells to beneficially use coal mine methane. Final report

    SciTech Connect

    Brown, J.T.; O`Brien, D.G.; Miller, A.R.; Atkins, R.; Sanders, M.

    1996-03-01

    DOE has been given the responsibility to encourage industry to recover and use methane that is currently being released to the atmosphere. At this time the only method being employed at the Left Fork Mine to remove methane is the mine ventilation system. The methane content was measured at one one-hundredth of a percent. To prevent this methane from being vented to the atmosphere, degasification wells are proposed. To use the coal mine methane, it is proposed to use phosphoric-acid fuel cells to convert methane to electric power. These fuel cells contain (1) a steam reformer to convert the methane to hydrogen (and carbon dioxide), (2) the fuel cell stack, and (3) a power conditioner that provides 200 kW of 60 Hz alternating current output. The environmental impacts and benefits of using this technology ware summarized in the report. The study indicates the methane emission reduction that could be achieved on a national and Global level. The important point being that this technology is economically viable as is demonstrated in the report.

  5. Technical Advance: Transcription factor, promoter, and enhancer utilization in human myeloid cells

    PubMed Central

    Joshi, Anagha; Pooley, Christopher; Freeman, Tom C.; Lennartsson, Andreas; Babina, Magda; Schmidl, Christian; Geijtenbeek, Teunis; Michoel, Tom; Severin, Jessica; Itoh, Masayoshi; Lassmann, Timo; Kawaji, Hideya; Hayashizaki, Yoshihide; Carninci, Piero; Forrest, Alistair R. R.; Rehli, Michael; Hume, David A.

    2015-01-01

    The generation of myeloid cells from their progenitors is regulated at the level of transcription by combinatorial control of key transcription factors influencing cell-fate choice. To unravel the global dynamics of this process at the transcript level, we generated transcription profiles for 91 human cell types of myeloid origin by use of CAGE profiling. The CAGE sequencing of these samples has allowed us to investigate diverse aspects of transcription control during myelopoiesis, such as identification of novel transcription factors, miRNAs, and noncoding RNAs specific to the myeloid lineage. We further reconstructed a transcription regulatory network by clustering coexpressed transcripts and associating them with enriched cis-regulatory motifs. With the use of the bidirectional expression as a proxy for enhancers, we predicted over 2000 novel enhancers, including an enhancer 38 kb downstream of IRF8 and an intronic enhancer in the KIT gene locus. Finally, we highlighted relevance of these data to dissect transcription dynamics during progressive maturation of granulocyte precursors. A multifaceted analysis of the myeloid transcriptome is made available (www.myeloidome.roslin.ed.ac.uk). This high-quality dataset provides a powerful resource to study transcriptional regulation during myelopoiesis and to infer the likely functions of unannotated genes in human innate immunity. PMID:25717144

  6. High Fuel Utilization in Solid Oxide Fuel Cells: Experimental Characterization and Data Analysis with Continuous Wavelet Transform

    NASA Astrophysics Data System (ADS)

    Esposito, Angelo; Russo, Luigi; Kändler, Christoph; Pianese, Cesare; Ludwig, Bastian; Steiner, Nadia Yousfi

    2016-06-01

    The on-line diagnostics of Solid Oxide Fuel Cells (SOFCs) is a critical tool to achieve optimal performance and extend the lifetime. The Continuous Wavelet Transform (CWT) methodology was applied to the SOFC voltage signal to detect signatures that reveal the presence of a fault in the cell/stack. The selected fault was anode re-oxidation caused by high Fuel Utilization (FU) (higher then nominal). To experimentally emulate the high FU faults, a standard test procedure was developed, which was used to characterize a μ-CHP system at high FU operation. To complete the analysis, data collected on Single Cells were exploited too. The CWT was applied to the voltage signal for each FU level to verify the qualitative difference (signature) between the signals at different FU's within the same tests as well as the correspondence between the same conditions over different tests. A statistical study was performed to quantify the observed differences and to determine the correspondence between CWT coefficients and operating conditions. The approach proves to be suitable to diagnose high FU in SOFC, showing a successful detection rate above 76%. The results show the good potential of using the CWT methodology as diagnostic tools for SOFCs from cell to stack level.

  7. [The presence of an endogenous peroxidase activity in hairy cell leukemia cells].

    PubMed

    Reyes, F; Gourdin, M F; Farcet, J P; Dreyfus, B; Breton-Gorius, J

    1977-02-01

    Mononuclear cells from hairy cell leukemia have been studied in three cases by ultrastructural immunocytochemistry. Cells have fairly detectable surface immunoglobulins, without monoclonal distribution however. In addition these cells have a peroxidatic activity which is revealed in the perinuclear space and strands of endoplasmic reticulum. PMID:404081

  8. Tissue engineered pre-vascularized buccal mucosa equivalents utilizing a primary triculture of epithelial cells, endothelial cells and fibroblasts.

    PubMed

    Heller, M; Frerick-Ochs, E V; Bauer, H-K; Schiegnitz, E; Flesch, D; Brieger, J; Stein, R; Al-Nawas, B; Brochhausen, C; Thüroff, J W; Unger, R E; Brenner, W

    2016-01-01

    Artificial generated buccal mucosa equivalents are a promising approach for the reconstruction of urethral defects. Limiting in this approach is a poor blood vessel supply after transplantation, resulting in increased morbidity and necrosis. We generated a pre-vascularized buccal mucosa equivalent in a tri-culture of primary buccal epithelial cells, fibroblasts and microvascular endothelial cells, using a native collagen membrane as a scaffold. A successful pre-vascularization and dense formation of capillary-like structures at superficial areas was demonstrated. The lumen size of pre-formed blood vessels corresponded to the capillary size in vivo (10-30 μm). Comparing native with a highly cross-linked collagen membrane we found a distinct higher formation of capillary-like structures on the native membrane, apparently caused by higher secretion of angiogenic factors such as PDGF, IL-8 and angiopoietin by the cells. These capillary-like structures became functional blood vessels through anastomosis with the host vasculature after implantation in nude mice. This in vitro method should result in an accelerated blood supply to the biomaterial with cells after transplantation and increase the succes rates of the implant material.

  9. Epstein-Barr Virus Utilizes Ikaros in Regulating Its Latent-Lytic Switch in B Cells

    PubMed Central

    Iempridee, Tawin; Reusch, Jessica A.; Riching, Andrew; Johannsen, Eric C.; Dovat, Sinisa; Kenney, Shannon C.

    2014-01-01

    ABSTRACT Ikaros is a zinc finger DNA-binding protein that regulates chromatin remodeling and the expression of genes involved in the cell cycle, apoptosis, and Notch signaling. It is a master regulator of lymphocyte differentiation and functions as a tumor suppressor in acute lymphoblastic leukemia. Nevertheless, no previous reports described effects of Ikaros on the life cycle of any human lymphotropic virus. Here, we demonstrate that full-length Ikaros (IK-1) functions as a major factor in the maintenance of viral latency in Epstein-Barr virus (EBV)-positive Burkitt's lymphoma Sal and MutuI cell lines. Either silencing of Ikaros expression by small hairpin RNA (shRNA) knockdown or ectopic expression of a non-DNA-binding isoform induced lytic gene expression. These effects synergized with other lytic inducers of EBV, including transforming growth factor β (TGF-β) and the hypoxia mimic desferrioxamine. Data from chromatin immunoprecipitation (ChIP)-quantitative PCR (qPCR) and ChIP-sequencing (ChIP-seq) analyses indicated that Ikaros did not bind to either of the EBV immediate early genes BZLF1 and BRLF1. Rather, Ikaros affected the expression of Oct-2 and Bcl-6, other transcription factors that directly inhibit EBV reactivation and plasma cell differentiation, respectively. IK-1 also complexed with the EBV immediate early R protein in coimmunoprecipitation assays and partially colocalized with R within cells. The presence of R alleviated IK-1-mediated transcriptional repression, with IK-1 then cooperating with Z and R to enhance lytic gene expression. Thus, we conclude that Ikaros plays distinct roles at different stages of EBV's life cycle: it contributes to maintaining latency via indirect mechanisms, and it may also synergize with Z and R to enhance lytic replication through direct association with R and/or R-induced alterations in Ikaros' functional activities via cellular signaling pathways. IMPORTANCE This is the first report showing that the cellular

  10. Benefits of utilizing CellProfiler as a characterization tool for U–10Mo nuclear fuel

    SciTech Connect

    Collette, R.; Douglas, J.; Patterson, L.; Bahun, G.; King, J.; Keiser, D.; Schulthess, J.

    2015-07-15

    Automated image processing techniques have the potential to aid in the performance evaluation of nuclear fuels by eliminating judgment calls that may vary from person-to-person or sample-to-sample. Analysis of in-core fuel performance is required for design and safety evaluations related to almost every aspect of the nuclear fuel cycle. This study presents a methodology for assessing the quality of uranium–molybdenum fuel images and describes image analysis routines designed for the characterization of several important microstructural properties. The analyses are performed in CellProfiler, an open-source program designed to enable biologists without training in computer vision or programming to automatically extract cellular measurements from large image sets. The quality metric scores an image based on three parameters: the illumination gradient across the image, the overall focus of the image, and the fraction of the image that contains scratches. The metric presents the user with the ability to ‘pass’ or ‘fail’ an image based on a reproducible quality score. Passable images may then be characterized through a separate CellProfiler pipeline, which enlists a variety of common image analysis techniques. The results demonstrate the ability to reliably pass or fail images based on the illumination, focus, and scratch fraction of the image, followed by automatic extraction of morphological data with respect to fission gas voids, interaction layers, and grain boundaries. - Graphical abstract: Display Omitted - Highlights: • A technique is developed to score U–10Mo FIB-SEM image quality using CellProfiler. • The pass/fail metric is based on image illumination, focus, and area scratched. • Automated image analysis is performed in pipeline fashion to characterize images. • Fission gas void, interaction layer, and grain boundary coverage data is extracted. • Preliminary characterization results demonstrate consistency of the algorithm.

  11. Utilizing Energy Transfer in Binary and Ternary Bulk Heterojunction Organic Solar Cells.

    PubMed

    Feron, Krishna; Cave, James M; Thameel, Mahir N; O'Sullivan, Connor; Kroon, Renee; Andersson, Mats R; Zhou, Xiaojing; Fell, Christopher J; Belcher, Warwick J; Walker, Alison B; Dastoor, Paul C

    2016-08-17

    Energy transfer has been identified as an important process in ternary organic solar cells. Here, we develop kinetic Monte Carlo (KMC) models to assess the impact of energy transfer in ternary and binary bulk heterojunction systems. We used fluorescence and absorption spectroscopy to determine the energy disorder and Förster radii for poly(3-hexylthiophene-2,5-diyl), [6,6]-phenyl-C61-butyric acid methyl ester, 4-bis[4-(N,N-diisobutylamino)-2,6-dihydroxyphenyl]squaraine (DIBSq), and poly(2,5-thiophene-alt-4,9-bis(2-hexyldecyl)-4,9-dihydrodithieno[3,2-c:3',2'-h][1,5]naphthyridine-5,10-dione). Heterogeneous energy transfer is found to be crucial in the exciton dissociation process of both binary and ternary organic semiconductor systems. Circumstances favoring energy transfer across interfaces allow relaxation of the electronic energy level requirements, meaning that a cascade structure is not required for efficient ternary organic solar cells. We explain how energy transfer can be exploited to eliminate additional energy losses in ternary bulk heterojunction solar cells, thus increasing their open-circuit voltage without loss in short-circuit current. In particular, we show that it is important that the DIBSq is located at the electron donor-acceptor interface; otherwise charge carriers will be trapped in the DIBSq domain or excitons in the DIBSq domains will not be able to dissociate efficiently at an interface. KMC modeling shows that only small amounts of DIBSq (<5% by weight) are needed to achieve substantial performance improvements due to long-range energy transfer. PMID:27456294

  12. Utilizing Energy Transfer in Binary and Ternary Bulk Heterojunction Organic Solar Cells.

    PubMed

    Feron, Krishna; Cave, James M; Thameel, Mahir N; O'Sullivan, Connor; Kroon, Renee; Andersson, Mats R; Zhou, Xiaojing; Fell, Christopher J; Belcher, Warwick J; Walker, Alison B; Dastoor, Paul C

    2016-08-17

    Energy transfer has been identified as an important process in ternary organic solar cells. Here, we develop kinetic Monte Carlo (KMC) models to assess the impact of energy transfer in ternary and binary bulk heterojunction systems. We used fluorescence and absorption spectroscopy to determine the energy disorder and Förster radii for poly(3-hexylthiophene-2,5-diyl), [6,6]-phenyl-C61-butyric acid methyl ester, 4-bis[4-(N,N-diisobutylamino)-2,6-dihydroxyphenyl]squaraine (DIBSq), and poly(2,5-thiophene-alt-4,9-bis(2-hexyldecyl)-4,9-dihydrodithieno[3,2-c:3',2'-h][1,5]naphthyridine-5,10-dione). Heterogeneous energy transfer is found to be crucial in the exciton dissociation process of both binary and ternary organic semiconductor systems. Circumstances favoring energy transfer across interfaces allow relaxation of the electronic energy level requirements, meaning that a cascade structure is not required for efficient ternary organic solar cells. We explain how energy transfer can be exploited to eliminate additional energy losses in ternary bulk heterojunction solar cells, thus increasing their open-circuit voltage without loss in short-circuit current. In particular, we show that it is important that the DIBSq is located at the electron donor-acceptor interface; otherwise charge carriers will be trapped in the DIBSq domain or excitons in the DIBSq domains will not be able to dissociate efficiently at an interface. KMC modeling shows that only small amounts of DIBSq (<5% by weight) are needed to achieve substantial performance improvements due to long-range energy transfer.

  13. Phase III Advanced Anodes and Cathodes Utilized in Energy Efficient Aluminum Production Cells

    SciTech Connect

    R.A. Christini; R.K. Dawless; S.P. Ray; D.A. Weirauch, Jr.

    2001-11-05

    During Phase I of the present program, Alcoa developed a commercial cell concept that has been estimated to save 30% of the energy required for aluminum smelting. Phase ii involved the construction of a pilot facility and operation of two pilots. Phase iii of the Advanced Anodes and Cathodes Program was aimed at bench experiments to permit the resolution of certain questions to be followed by three pilot cells. All of the milestones related to materials, in particular metal purity, were attained with distinct improvements over work in previous phases of the program. NiO additions to the ceramic phase and Ag additions to the Cu metal phase of the cermet improved corrosion resistance sufficiently that the bench scale pencil anodes met the purity milestones. Some excellent metal purity results have been obtained with anodes of the following composition: Further improvements in anode material composition appear to be dependent on a better understanding of oxide solubilities in molten cryolite. For that reason, work was commissioned with an outside consultant to model the MeO - cryolite systems. That work has led to a better understanding of which oxides can be used to substitute into the NiO-Fe2O3 ceramic phase to stabilize the ferrites and reduce their solubility in molten cryolite. An extensive number of vertical plate bench electrolysis cells were run to try to find conditions where high current efficiencies could be attained. TiB2-G plates were very inconsistent and led to poor wetting and drainage. Pure TiB2 did produce good current efficiencies at small overlaps (shadowing) between the anodes and cathodes. This bench work with vertical plate anodes and cathodes reinforced the importance of good cathode wetting to attain high current efficiencies. Because of those conclusions, new wetting work was commissioned and became a major component of the research during the third year of Phase III. While significant progress was made in several areas, much work needs to be

  14. Circulating Tumor Cells and Circulating Tumor DNA: Challenges and Opportunities on the Path to Clinical Utility.

    PubMed

    Ignatiadis, Michail; Lee, Mark; Jeffrey, Stefanie S

    2015-11-01

    Recent technological advances have enabled the detection and detailed characterization of circulating tumor cells (CTC) and circulating tumor DNA (ctDNA) in blood samples from patients with cancer. Often referred to as a "liquid biopsy," CTCs and ctDNA are expected to provide real-time monitoring of tumor evolution and therapeutic efficacy, with the potential for improved cancer diagnosis and treatment. In this review, we focus on these opportunities as well as the challenges that should be addressed so that these tools may eventually be implemented into routine clinical care.

  15. Benefits of utilizing CellProfiler as a characterization tool for U-10Mo nuclear fuel

    DOE PAGESBeta

    Collette, R.; Douglas, J.; Patterson, L.; Bahun, G.; King, J.; Keiser, D.; Schulthess, J.

    2015-05-01

    Automated image processing techniques have the potential to aid in the performance evaluation of nuclear fuels by eliminating judgment calls that may vary from person-to-person or sample-to-sample. Analysis of in-core fuel performance is required for design and safety evaluations related to almost every aspect of the nuclear fuel cycle. This study presents a methodology for assessing the quality of uranium-molybdenum fuel images and describes image analysis routines designed for the characterization of several important microstructural properties. The analyses are performed in CellProfiler, an open-source program designed to enable biologists without training in computer vision or programming to automatically extract cellularmore » measurements from large image sets. The quality metric scores an image based on three parameters: the illumination gradient across the image, the overall focus of the image, and the fraction of the image that contains scratches. The metric presents the user with the ability to ‘pass’ or ‘fail’ an image based on a reproducible quality score. Passable images may then be characterized through a separate CellProfiler pipeline, which enlists a variety of common image analysis techniques. The results demonstrate the ability to reliably pass or fail images based on the illumination, focus, and scratch fraction of the image, followed by automatic extraction of morphological data with respect to fission gas voids, interaction layers, and grain boundaries.« less

  16. Benefits of utilizing CellProfiler as a characterization tool for U-10Mo nuclear fuel

    SciTech Connect

    Collette, R.; Douglas, J.; Patterson, L.; Bahun, G.; King, J.; Keiser, D.; Schulthess, J.

    2015-05-01

    Automated image processing techniques have the potential to aid in the performance evaluation of nuclear fuels by eliminating judgment calls that may vary from person-to-person or sample-to-sample. Analysis of in-core fuel performance is required for design and safety evaluations related to almost every aspect of the nuclear fuel cycle. This study presents a methodology for assessing the quality of uranium-molybdenum fuel images and describes image analysis routines designed for the characterization of several important microstructural properties. The analyses are performed in CellProfiler, an open-source program designed to enable biologists without training in computer vision or programming to automatically extract cellular measurements from large image sets. The quality metric scores an image based on three parameters: the illumination gradient across the image, the overall focus of the image, and the fraction of the image that contains scratches. The metric presents the user with the ability to ‘pass’ or ‘fail’ an image based on a reproducible quality score. Passable images may then be characterized through a separate CellProfiler pipeline, which enlists a variety of common image analysis techniques. The results demonstrate the ability to reliably pass or fail images based on the illumination, focus, and scratch fraction of the image, followed by automatic extraction of morphological data with respect to fission gas voids, interaction layers, and grain boundaries.

  17. Anode catalysts for direct ethanol fuel cells utilizing directly solar light illumination.

    PubMed

    Chu, Daobao; Wang, Shuxi; Zheng, Peng; Wang, Jian; Zha, Longwu; Hou, Yuanyuan; He, Jianguo; Xiao, Ying; Lin, Huashui; Tian, Zhaowu

    2009-01-01

    Shine a light: A PtNiRu/TiO(2) anode catalyst for direct ethanol fuel cells shows photocatalytic activity. The peak current density for ethanol oxidation under solar light illumination is 2-3 times greater than that in the absence of solar light. Ethanol is oxidized by light-generated holes, and the electrons are collected by the TiO(2) support to generate the oxidation current.Novel PtNiRu/TiO(2) anode catalysts for direct ethanol fuel cells (DEFCs) were prepared from PtNiRu nanoparticles (1:1:1 atomic ratios) and a nanoporous TiO(2) film by a sol-gel and electrodeposition method. The performances of the catalysts for ethanol oxidation were investigated by cyclic voltammetry, chronoamperometry and electrochemical impedance spectroscopy. The results indicate a remarkable enhancement of activity for ethanol oxidation under solar light illumination. Under solar light illumination, the generated oxidation peak current density is 24.6 mA cm(-2), which is about 2.5 times higher than that observed without solar light (9.9 mA cm(-2)). The high catalytic activity of the PtNiRu/TiO(2) complex catalyst for the electrooxidation of ethanol may be attributed to the modified metal/nanoporous TiO(2) film, and the enhanced electrooxidation of ethanol under solar light may be due to the photogeneration of holes in the modified nanoporous TiO(2) film.

  18. Enhanced conversion efficiency in perovskite solar cells by effectively utilizing near infrared light.

    PubMed

    Que, Meidan; Que, Wenxiu; Yin, Xingtian; Chen, Peng; Yang, Yawei; Hu, Jiaxing; Yu, Boyan; Du, Yaping

    2016-08-14

    Up-conversion β-NaYF4:Yb(3+),Tm(3+)/NaYF4 core-shell nanoparticles (NYF NPs) with a high luminous intensity in the visible light region were synthesized by a hydrothermal reaction process. Photocurrent densities of the mesoscopic perovskite solar cells fabricated by incorporating up-conversion NYF NPs into the electron transporting layer are effectively enhanced. The effects of the thicknesses of the electron transporting layer and the weight ratio of up-conversion NYF NPs/TiO2 on the power conversion efficiency (PCE) of the as-fabricated devices were also investigated. The results indicate that the PCE of the optimized device achieves 16.9%, which is 20% higher than that of the device without introducing NYF NPs, and the steady-state PCE of the as-fabricated devices is close to its transient-state PCE. The up-conversion effect of NYF NPs is conducive to higher device performance rather than the nanoparticles as scattering centers to increase possible light absorption of the perovskite film or the electronic effect of the NaYF4 shell surface. These results can be further confirmed by finite-difference time-domain simulation. Photoluminescence results suggest that the multiphonon-assistance can accelerate the nonradiative recombination process at a lower temperature. Incorporating NYF NPs into the electron transporting layer opens a new approach to a promising family of electron transporting materials for mesoscopic perovskite solar cells. PMID:27406678

  19. Platform for a hydrocarbon exhaust gas sensor utilizing a pumping cell and a conductometric sensor.

    PubMed

    Biskupski, Diana; Geupel, Andrea; Wiesner, Kerstin; Fleischer, Maximilian; Moos, Ralf

    2009-01-01

    Very often, high-temperature operated gas sensors are cross-sensitive to oxygen and/or they cannot be operated in oxygen-deficient (rich) atmospheres. For instance, some metal oxides like Ga(2)O(3) or doped SrTiO(3) are excellent materials for conductometric hydrocarbon detection in the rough atmosphere of automotive exhausts, but have to be operated preferably at a constant oxygen concentration. We propose a modular sensor platform that combines a conductometric two-sensor-setup with an electrochemical pumping cell made of YSZ to establish a constant oxygen concentration in the ambient of the conductometric sensor film. In this paper, the platform is introduced, the two-sensor-setup is integrated into this new design, and sensing performance is characterized. Such a platform can be used for other sensor principles as well. PMID:22423212

  20. Enhanced conversion efficiency in perovskite solar cells by effectively utilizing near infrared light

    NASA Astrophysics Data System (ADS)

    Que, Meidan; Que, Wenxiu; Yin, Xingtian; Chen, Peng; Yang, Yawei; Hu, Jiaxing; Yu, Boyan; Du, Yaping

    2016-07-01

    Up-conversion β-NaYF4:Yb3+,Tm3+/NaYF4 core-shell nanoparticles (NYF NPs) with a high luminous intensity in the visible light region were synthesized by a hydrothermal reaction process. Photocurrent densities of the mesoscopic perovskite solar cells fabricated by incorporating up-conversion NYF NPs into the electron transporting layer are effectively enhanced. The effects of the thicknesses of the electron transporting layer and the weight ratio of up-conversion NYF NPs/TiO2 on the power conversion efficiency (PCE) of the as-fabricated devices were also investigated. The results indicate that the PCE of the optimized device achieves 16.9%, which is 20% higher than that of the device without introducing NYF NPs, and the steady-state PCE of the as-fabricated devices is close to its transient-state PCE. The up-conversion effect of NYF NPs is conducive to higher device performance rather than the nanoparticles as scattering centers to increase possible light absorption of the perovskite film or the electronic effect of the NaYF4 shell surface. These results can be further confirmed by finite-difference time-domain simulation. Photoluminescence results suggest that the multiphonon-assistance can accelerate the nonradiative recombination process at a lower temperature. Incorporating NYF NPs into the electron transporting layer opens a new approach to a promising family of electron transporting materials for mesoscopic perovskite solar cells.Up-conversion β-NaYF4:Yb3+,Tm3+/NaYF4 core-shell nanoparticles (NYF NPs) with a high luminous intensity in the visible light region were synthesized by a hydrothermal reaction process. Photocurrent densities of the mesoscopic perovskite solar cells fabricated by incorporating up-conversion NYF NPs into the electron transporting layer are effectively enhanced. The effects of the thicknesses of the electron transporting layer and the weight ratio of up-conversion NYF NPs/TiO2 on the power conversion efficiency (PCE) of the as

  1. Platform for a Hydrocarbon Exhaust Gas Sensor Utilizing a Pumping Cell and a Conductometric Sensor

    PubMed Central

    Biskupski, Diana; Geupel, Andrea; Wiesner, Kerstin; Fleischer, Maximilian; Moos, Ralf

    2009-01-01

    Very often, high-temperature operated gas sensors are cross-sensitive to oxygen and/or they cannot be operated in oxygen-deficient (rich) atmospheres. For instance, some metal oxides like Ga2O3 or doped SrTiO3 are excellent materials for conductometric hydrocarbon detection in the rough atmosphere of automotive exhausts, but have to be operated preferably at a constant oxygen concentration. We propose a modular sensor platform that combines a conductometric two-sensor-setup with an electrochemical pumping cell made of YSZ to establish a constant oxygen concentration in the ambient of the conductometric sensor film. In this paper, the platform is introduced, the two-sensor-setup is integrated into this new design, and sensing performance is characterized. Such a platform can be used for other sensor principles as well. PMID:22423212

  2. Development of a Solid Oxide Fuel Cell for the utilization of coal mine gas

    NASA Astrophysics Data System (ADS)

    Groß, B.; Blum, L.; de Haart, L. G. J.; Dengel, A.

    Apart from natural gas there is another important natural source of methane. The so-called coal mine gas is a by-product of the geochemical process of the carbonization of sediments from marsh woods of the Earth's Carboniferous Period. Methane evaporates from the coal and has to be removed out of the active mines where it represents one of the main safety risks. Methane also evaporates in abandoned coal mines. In the federal state Saarland in Germany exists above ground a more than 110 km pipeline for the drained coal mine gas from 12 different sources. The content of methane varies between 25 and 90%, the oxygen content (from air) is in the range up to 10%. This wide range or variation, respectively, of fuel and oxygen content, causes a lot of problems for the use in conventional engines. Therefore the company Evonik New Energies GmbH is interested in using SOFC with coal mine gas as efficient as possible to produce electric power. For that purpose at Forschungszentrum Jülich the available SOFC technology was adapted to the use with coal mine gas and a test facility was designed to operate an SOFC stack (approximately 2 kW electrical power output) together with a pre-reformer. This paper presents the results of the coal mine gas analysis and the effect on the pre-reformer and the fuel cell. The composition of the coal mine gas was determined by means of micro-gas chromatography. The results obtained from preliminary tests using synthetic and real coal mine gas on the pre-reformer and on the fuel cell are discussed.

  3. Perspectives on utilizing unique features of microfluidics technology for particle and cell sorting

    PubMed Central

    Adams, Jonathan D.; Tom Soh, H.

    2009-01-01

    Sample preparation is often the most tedious and demanding step in an assay, but it also plays an essential role in determining the quality of results. As biological questions and analytical methods become increasingly sophisticated, there is a rapidly growing need for systems that can reliably and reproducibly separate cells and particles with high purity, throughput and recovery. Microfluidics technology represents a compelling approach in this regard, allowing precise control of separation forces for high performance separation in inexpensive, or even disposable, devices. In addition, microfluidics technology enables the fabrication of arrayed and integrated systems that operate either in parallel or in tandem, in a capacity that would be difficult to achieve in macro-scale systems. In this report, we use recent examples from our work to illustrate the potential of microfluidic cell- and particle-sorting devices. We demonstrate the potential of chip-based high-gradient magnetophoresis that enable high-purity separation through reversible trapping of target particles paired with high-stringency washing with minimal loss. We also describe our work in the development of devices that perform simultaneous multi-target sorting, either through precise control of magnetic and fluidic forces or through the integration of multiple actuation forces into a single monolithic device. We believe that such devices may serve as a powerful “front-end” module of highly integrated analytical platforms capable of providing actionable diagnostic information directly from crude, unprocessed samples - the success of such systems may hold the key to advancing point-of-care diagnostics and personalized medicine. PMID:20161387

  4. Ceria coated Ni as anodes for direct utilization of methane in low-temperature solid oxide fuel cells

    NASA Astrophysics Data System (ADS)

    Zhu, Wei; Xia, Changrong; Fan, Jue; Peng, Ranran; Meng, Guangyao

    A new type of anode, a Ni framework coated with Sm-doped ceria (SDC), was developed for direct utilization of methane fuel in low-temperature solid oxide fuel cells (SOFCs) with thin-film SDC electrolytes. The coated SDC was prepared with an ion impregnating method and the electrolyte films were fabricated with a co-pressing and co-firing technique. The impregnating process produced an ideal anode microstructure where nickel particles were effectively connected and uniformly covered with nanosized SDC. This anode microstructure was believed to enlarge the triple-phase boundaries and therefore enhance the anode performance. The cell performance was much higher than that of a conventional fuel cell with a Ni-SDC composite anode. In addition, the performance increased with impregnated SDC loading up to a maximum at 20 mg cm -2, indicating that the coated SDC is the contributing factor for the enhanced fuel cell performance. Power density as high as 571 and 353 mW cm -2 were obtained at 600 °C when humidified hydrogen and methane were used as fuels, respectively. The stability of the cell also increased with the SDC loading. No significant degradation was observed for anodes coated with 20 and 25 mg cm -2 SDC. This verifies that the coated SDC electrodes are very effective in suppressing catalytic carbon formation by blocking methane from approaching the Ni, which is catalytically active towards methane pyrolysis. The high performance of this anode shows high promise in the developing field of direct hydrocarbon SOFCs.

  5. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions

    PubMed Central

    Fraley, Stephanie I.; Wu, Pei-hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D.; Wirtz, Denis

    2015-01-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility. PMID:26423227

  6. Strategies for the production of cell wall-deconstructing enzymes in lignocellulosic biomass and their utilization for biofuel production.

    PubMed

    Park, Sang-Hyuck; Ong, Rebecca Garlock; Sticklen, Mariam

    2016-06-01

    Microbial cell wall-deconstructing enzymes are widely used in the food, wine, pulp and paper, textile, and detergent industries and will be heavily utilized by cellulosic biorefineries in the production of fuels and chemicals. Due to their ability to use freely available solar energy, genetically engineered bioenergy crops provide an attractive alternative to microbial bioreactors for the production of cell wall-deconstructing enzymes. This review article summarizes the efforts made within the last decade on the production of cell wall-deconstructing enzymes in planta for use in the deconstruction of lignocellulosic biomass. A number of strategies have been employed to increase enzyme yields and limit negative impacts on plant growth and development including targeting heterologous enzymes into specific subcellular compartments using signal peptides, using tissue-specific or inducible promoters to limit the expression of enzymes to certain portions of the plant or certain times, and fusion of amplification sequences upstream of the coding region to enhance expression. We also summarize methods that have been used to access and maintain activity of plant-generated enzymes when used in conjunction with thermochemical pretreatments for the production of lignocellulosic biofuels. PMID:26627868

  7. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions.

    PubMed

    Fraley, Stephanie I; Wu, Pei-Hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D; Wirtz, Denis

    2015-01-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility. PMID:26423227

  8. Cell Phone Utilization among Foreign-born Latinos: a Promising Tool for Dissemination of Health and HIV Information

    PubMed Central

    Leite, Lorena; Buresh, Megan; Rios, Naomi; Conley, Anna; Flys, Tamara; Page, Kathleen R.

    2013-01-01

    Latinos in the U.S. are disproportionately affected by HIV and are at risk for late presentation to care. Between June 2011 and January 2012, we conducted a cross-sectional survey of 209 Baltimore Latinos at community-based venues to evaluate the feasibility of using communication technology (ICT)-based interventions to improve access to HIV testing and education within the Spanish-speaking community in Baltimore. Participants had a median age of 33 years (IQR 28-42), 51.7% were male, and 95.7% were foreign-born. Approximately two-thirds (63.2%) had been in the U.S. less than 10 years and 70.1% had been previously tested for HIV. Cell phone (92.3%) and text messaging (74.2%) was used more than Internet (52.2%) or e-mail (42.8%) (p<0.01). In multivariate analysis, older age and lower education were associated with less utilization of Internet, e-mail and text messaging, but not cell phones. Interest was high for receiving health education (73.1%), HIV education (70.2%), and test results (68.8%) via text messaging. Innovative cell phone-based communication interventions have the potential to link Latino migrants to HIV prevention, testing and treatment services. PMID:23440452

  9. Strategies for the production of cell wall-deconstructing enzymes in lignocellulosic biomass and their utilization for biofuel production.

    PubMed

    Park, Sang-Hyuck; Ong, Rebecca Garlock; Sticklen, Mariam

    2016-06-01

    Microbial cell wall-deconstructing enzymes are widely used in the food, wine, pulp and paper, textile, and detergent industries and will be heavily utilized by cellulosic biorefineries in the production of fuels and chemicals. Due to their ability to use freely available solar energy, genetically engineered bioenergy crops provide an attractive alternative to microbial bioreactors for the production of cell wall-deconstructing enzymes. This review article summarizes the efforts made within the last decade on the production of cell wall-deconstructing enzymes in planta for use in the deconstruction of lignocellulosic biomass. A number of strategies have been employed to increase enzyme yields and limit negative impacts on plant growth and development including targeting heterologous enzymes into specific subcellular compartments using signal peptides, using tissue-specific or inducible promoters to limit the expression of enzymes to certain portions of the plant or certain times, and fusion of amplification sequences upstream of the coding region to enhance expression. We also summarize methods that have been used to access and maintain activity of plant-generated enzymes when used in conjunction with thermochemical pretreatments for the production of lignocellulosic biofuels.

  10. Cell phone utilization among foreign-born Latinos: a promising tool for dissemination of health and HIV information.

    PubMed

    Leite, Lorena; Buresh, Megan; Rios, Naomi; Conley, Anna; Flys, Tamara; Page, Kathleen R

    2014-08-01

    Latinos in the US are disproportionately affected by HIV and are at risk for late presentation to care. Between June 2011 and January 2012, we conducted a cross-sectional survey of 209 Baltimore Latinos at community-based venues to evaluate the feasibility of using information communication technology-based interventions to improve access to HIV testing and education within the Spanish-speaking community in Baltimore. Participants had a median age of 33 years interquartile range (IQR) (IQR 28-42), 51.7 % were male, and 95.7 % were foreign-born. Approximately two-thirds (63.2 %) had been in the US less than 10 years and 70.1 % had been previously tested for HIV. Cell phone (92.3 %) and text messaging (74.2 %) was used more than Internet (52.2 %) or e-mail (42.8 %) (p < 0.01). In multivariate analysis, older age and lower education were associated with less utilization of Internet, e-mail and text messaging, but not cell phones. Interest was high for receiving health education (73.1 %), HIV education (70.2 %), and test results (68.8 %) via text messaging. Innovative cell phone-based communication interventions have the potential to link Latino migrants to HIV prevention, testing and treatment services.

  11. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions

    NASA Astrophysics Data System (ADS)

    Fraley, Stephanie I.; Wu, Pei-Hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D.; Wirtz, Denis

    2015-10-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility.

  12. Utility of Iron Staining in Identifying the Cause of Renal Allograft Dysfunction in Patients with Sickle Cell Disease.

    PubMed

    Wang, Yingchun; Doshi, Mona; Khan, Salman; Li, Wei; Zhang, Ping L

    2015-01-01

    Sickle cell nephropathy (SCN) is associated with iron/heme deposition in proximal renal tubules and related acute tubular injury (ATI). Here we report the utility of iron staining in differentiating causes of renal allograft dysfunction in patients with a history of sickle cell disease. Case 1: the patient developed acute allograft dysfunction two years after renal transplant. Her renal biopsy showed ATI, supported by patchy loss of brush border and positive staining of kidney injury molecule-1 in proximal tubular epithelial cells, where diffuse increase in iron staining (2+) was present. This indicated that ATI likely resulted from iron/heme toxicity to proximal tubules. Electron microscope confirmed aggregated sickle RBCs in glomeruli, indicating a recurrent SCN. Case 2: four years after renal transplant, the patient developed acute allograft dysfunction and became positive for serum donor-specific antibody. His renal biopsy revealed thrombotic microangiopathy (TMA) and diffuse positive C4d stain in peritubular capillaries. Iron staining was negative in the renal tubules, implying that TMA was likely associated with acute antibody-mediated rejection (AAMR, type 2) rather than recurrent SCN. These case reports imply that iron staining is an inexpensive but effective method in distinguishing SCN-associated renal injury in allograft kidney from other etiologies. PMID:26697257

  13. Membrane/mediator-free rechargeable enzymatic biofuel cell utilizing graphene/single-wall carbon nanotube cogel electrodes.

    PubMed

    Campbell, Alan S; Jeong, Yeon Joo; Geier, Steven M; Koepsel, Richard R; Russell, Alan J; Islam, Mohammad F

    2015-02-25

    Enzymatic biofuel cells (EBFCs) utilize enzymes to convert chemical energy present in renewable biofuels into electrical energy and have shown much promise in the continuous powering of implantable devices. Currently, however, EBFCs are greatly limited in terms of power and operational stability with a majority of reported improvements requiring the inclusion of potentially toxic and unstable electron transfer mediators or multicompartment systems separated by a semipermeable membrane resulting in complicated setups. We report on the development of a simple, membrane/mediator-free EBFC utilizing novel electrodes of graphene and single-wall carbon nanotube cogel. These cogel electrodes had large surface area (∼ 800 m(2) g(-1)) that enabled high enzyme loading, large porosity for unhindered glucose transport and moderate electrical conductivity (∼ 0.2 S cm(-1)) for efficient charge collection. Glucose oxidase and bilirubin oxidase were physically adsorbed onto these electrodes to form anodes and cathodes, respectively, and the EBFC produced power densities up to 0.19 mW cm(-2) that correlated to 0.65 mW mL(-1) or 140 mW g(-1) of GOX with an open circuit voltage of 0.61 V. Further, the electrodes were rejuvenated by a simple wash and reloading procedure. We postulate these porous and ultrahigh surface area electrodes will be useful for biosensing applications, and will allow reuse of EBFCs.

  14. Utilization of the Generalized Method of Cells to Analyze the Deformation Response of Laminated Ceramic Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.

    2012-01-01

    In order to practically utilize ceramic matrix composites in aircraft engine components, robust analysis tools are required that can simulate the material response in a computationally efficient manner. The MAC/GMC software developed at NASA Glenn Research Center, based on the Generalized Method of Cells micromechanics method, has the potential to meet this need. Utilizing MAC/GMC, the effective stiffness properties, proportional limit stress and ultimate strength can be predicted based on the properties and response of the individual constituents. In this paper, the effective stiffness and strength properties for a representative laminated ceramic matrix composite with a large diameter fiber are predicted for a variety of fiber orientation angles and laminate orientations. As part of the analytical study, methods to determine the in-situ stiffness and strength properties of the constituents required to appropriately simulate the effective composite response are developed. The stiffness properties of the representative composite have been adequately predicted for all of the fiber orientations and laminate configurations examined in this study. The proportional limit stresses and strains and ultimate stresses and strains were predicted with varying levels of accuracy, depending on the laminate orientation. However, for the cases where the predictions did not have the desired level of accuracy, the specific issues related to the micromechanics theory were identified which could lead to difficulties that were encountered that could be addressed in future work.

  15. The insect endosymbiont Sodalis glossinidius utilizes a type III secretion system for cell invasion.

    PubMed

    Dale, C; Young, S A; Haydon, D T; Welburn, S C

    2001-02-13

    Sodalis glossinidius is a maternally transmitted secondary endosymbiont residing intracellularly in tissues of the tsetse flies, Glossina spp. In this study, we have used Tn5 mutagenesis and a negative selection procedure to derive a S. glossinidius mutant that is incapable of invading insect cells in vitro and is aposymbiotic when microinjected into tsetse. This mutant strain harbors Tn5 integrated into a chromosomal gene sharing high sequence identity with a type III secretion system invasion gene (invC) previously identified in Salmonella enterica. With the use of degenerate PCR, we have amplified a further six Sodalis inv/spa genes sharing high sequence identity with type III secretion system genes encoded by Salmonella pathogenicity island 1. Phylogenetic reconstructions based on the inv/spa genes of Sodalis and other members of the family Enterobacteriaceae have consistently identified a well-supported clade containing Sodalis and the enteric pathogens Shigella and Salmonella. These results suggest that Sodalis may have evolved from an ancestor with a parasitic intracellular lifestyle, possibly a latter-day entomopathogen. These observations lend credence to a hypothesis suggesting that vertically transmitted mutualistic endosymbionts evolve from horizontally transmitted parasites through a parasitism-mutualism continuum.

  16. Heat removal from high temperature tubular solid oxide fuel cells utilizing product gas from coal gasifiers.

    SciTech Connect

    Parkinson, W. J. ,

    2003-01-01

    In this work we describe the results of a computer study used to investigate the practicality of several heat exchanger configurations that could be used to extract heat from tubular solid oxide fuel cells (SOFCs) . Two SOFC feed gas compositions were used in this study. They represent product gases from two different coal gasifier designs from the Zero Emission Coal study at Los Alamos National Laboratory . Both plant designs rely on the efficient use of the heat produced by the SOFCs . Both feed streams are relatively rich in hydrogen with a very small hydrocarbon content . One feed stream has a significant carbon monoxide content with a bit less hydrogen . Since neither stream has a significant hydrocarbon content, the common use of the endothermic reforming reaction to reduce the process heat is not possible for these feed streams . The process, the method, the computer code, and the results are presented as well as a discussion of the pros and cons of each configuration for each process .

  17. Potential therapeutic utility of mesenchymal stem cells in inflammatory bowel disease in mice.

    PubMed

    Abdel Salam, Ahmed G; Ata, Hazem M; Salman, Tarek M; Rashed, Laila A; Sabry, Dina; Schaalan, Mona F

    2014-10-01

    Mesenchymal stem cells (MSCs) were found to provide an effective therapeutic role in inflammatory diseases by modulating inflammatory responses and tissue regeneration by their differentiation ability. The present work sought to demonstrate the potential therapeutic use of MSCs in treating chronic inflammatory bowel disease (IBD) in mice. A new model to induce chronic IBD based on alternative administration periods of Dextran Sodium Sulfate (DSS) was established. Mice were divided into 2 groups; one was treated with MSCs and the other was treated with phosphate-buffered saline (PBS). Assessment of therapeutic efficacy of MSCs was by measuring weight, stool scoring, histopathological examination, and measuring the gene expression of inflammatory markers: Interleukin-23 (IL-23), Tumor necrosis factor-α (TNF-α), Interferon-γ (IFN-γ), and Intercellular adhesion molecule-1 (ICAM-1). The results showed that DSS administration causes bloody and watery stool, weight loss, and altered histopathologic picture. MSC treated mice showed a significant improvement in stool condition, weight gain, and normal histopathologic picture compared to the PBS treated mice. Moreover, gene expressions of inflammatory markers in the intestines of the MSC treated mice were also significantly lower than those of the PBS treated mice. In conclusion, the data here showed that MSCs have a clear potential efficacy in the treatment for IBD, as their immune modulation effects include inhibition in the expression of key inflammatory markers that each plays an important role in the pathogenesis of IBD.

  18. The insect endosymbiont Sodalis glossinidius utilizes a type III secretion system for cell invasion

    PubMed Central

    Dale, Colin; Young, Simon A.; Haydon, Daniel T.; Welburn, Susan C.

    2001-01-01

    Sodalis glossinidius is a maternally transmitted secondary endosymbiont residing intracellularly in tissues of the tsetse flies, Glossina spp. In this study, we have used Tn5 mutagenesis and a negative selection procedure to derive a S. glossinidius mutant that is incapable of invading insect cells in vitro and is aposymbiotic when microinjected into tsetse. This mutant strain harbors Tn5 integrated into a chromosomal gene sharing high sequence identity with a type III secretion system invasion gene (invC) previously identified in Salmonella enterica. With the use of degenerate PCR, we have amplified a further six Sodalis inv/spa genes sharing high sequence identity with type III secretion system genes encoded by Salmonella pathogenicity island 1. Phylogenetic reconstructions based on the inv/spa genes of Sodalis and other members of the family Enterobacteriaceae have consistently identified a well-supported clade containing Sodalis and the enteric pathogens Shigella and Salmonella. These results suggest that Sodalis may have evolved from an ancestor with a parasitic intracellular lifestyle, possibly a latter-day entomopathogen. These observations lend credence to a hypothesis suggesting that vertically transmitted mutualistic endosymbionts evolve from horizontally transmitted parasites through a parasitism–mutualism continuum. PMID:11172045

  19. Hematopoietic stem and progenitor cell harvesting: technical advances and clinical utility

    PubMed Central

    Hequet, Olivier

    2015-01-01

    Hematopoietic stem and progenitor cell (HSPC) transplantations require prior harvesting of allogeneic or autologous HSPCs. HSPCs are usually present in bone marrow (BM) during the entire life, in cord blood (CB) at birth, or in peripheral blood (PB) under particular circumstances. HSPCs were first harvested in BM and later in CB and PB, as studies showed interesting features of such grafts. All harvesting methods were in use throughout the years, except BM harvesting for HSPC autologous transplantation, which was replaced by PB harvesting. BM, CB, and PB harvesting methods have been developed, and materials and devices technically improved to increase the number of HSPCs harvested. In parallel, knowing the features of the donors or patients associated with successful numbers of HSPCs allows the adaptation of appropriate harvesting methods. Moreover, it is important to ensure the safety of donors or patients while harvesting. This review describes the methods used for harvesting based on recent studies or developments around these methods, and more particularly, the means developed to increase the numbers of HSPCs harvested in each method. It also explains briefly the influence of technical improvements in HSPC harvesting on potential changes in HSPC graft composition. PMID:25733943

  20. Utility of saliva and hair follicles in donor selection for hematopoietic stem cell transplantation and chimerism monitoring.

    PubMed

    Kaur, Gurvinder; Kumar, Neeraj; Nandakumar, Ramya; Rapthap, Chowphi C; Sharma, Gaurav; Neolia, Shekhar; Kumra, Heena; Mahalwar, Prateek; Garg, Abhinav; Kumar, Sunil; Kaur, Jasmeet; Hakim, Mrinali; Kumar, Lalit; Mehra, Narinder K

    2012-01-01

    Selection of an HLA identical donor is a critical pre-requisite for successful hematopoietic stem cell transplantation (HSCT). Most transplant centers utilize blood as the most common source of DNA for HLA testing. However, obtaining blood through phlebotomy is often challenging in patients with conditions like severe leucopenia or hemophilia, pediatric and elderly patients. We have used a simple in-house protocol and shown that HLA genotypes obtained on DNA extracted from saliva or hair are concordant with blood and hence can be used for selection of donors for HSCT or organ transplantation. Similarly, for post-HSCT chimerism monitoring, non-availability of pre-transplant DNA samples poses a major limitation of reference STR fingerprints. This study shows that DNA obtained post-HSCT from hair follicles can be used to generate pre-transplant patient specific fingerprints while the STR profiles obtained in saliva samples cannot as these display a mixed state of chimerism.

  1. Highly efficient colorimetric detection of target cancer cells utilizing superior catalytic activity of graphene oxide-magnetic-platinum nanohybrids

    NASA Astrophysics Data System (ADS)

    Kim, Moon Il; Kim, Min Su; Woo, Min-Ah; Ye, Youngjin; Kang, Kyoung Suk; Lee, Jinwoo; Park, Hyun Gyu

    2014-01-01

    Enzyme-linked immunosorbent assays (ELISAs) have most widely been applied in immunoassays for several decades. However, several unavoidable limitations (e.g., instability caused by structural unfolding) of natural enzymes have hindered their widespread applications. Here, we describe a new nanohybrid consisting of Fe3O4 magnetic nanoparticles (MNPs) and platinum nanoparticles (Pt NPs), simultaneously immobilized on the surface of graphene oxide (GO). By synergistically integrating highly catalytically active Pt NPs and MNPs on GO whose frameworks possess high substrate affinity, the nanohybrid is able to achieve up to a 30-fold higher maximal reaction velocity (Vmax) compared to that of free GO for the colorimetric reaction of the peroxidase substrate, 3,3',5,5'-tetramethylbenzidine (TMB), and enable rapid detection of target cancer cells. Specifically, using this new assay system, clinically important breast cancer cells are detected in a 5 min time period at room temperature with high specificity and sensitivity. The remarkably high capability to catalyze oxidation reactions could allow the nanohybrid to replace conventional peroxidase-based immunoassay systems as part of new, rapid, robust and convenient assay systems which can be widely utilized for the identification of important target molecules.Enzyme-linked immunosorbent assays (ELISAs) have most widely been applied in immunoassays for several decades. However, several unavoidable limitations (e.g., instability caused by structural unfolding) of natural enzymes have hindered their widespread applications. Here, we describe a new nanohybrid consisting of Fe3O4 magnetic nanoparticles (MNPs) and platinum nanoparticles (Pt NPs), simultaneously immobilized on the surface of graphene oxide (GO). By synergistically integrating highly catalytically active Pt NPs and MNPs on GO whose frameworks possess high substrate affinity, the nanohybrid is able to achieve up to a 30-fold higher maximal reaction velocity (Vmax

  2. Utility of Red Cell Distribution Width as a Prognostic Factor in Young Breast Cancer Patients

    PubMed Central

    Huang, Du-Ping; Ma, Rui-Min; Xiang, You-Qun

    2016-01-01

    Abstract The prognosis of breast cancer occurs in young women is usually poor. Red cell distribution width (RDW), 1 of many routinely examined parameters, has recently been proposed as a prognostic marker in solid tumors. The aim of our study was to assess the predictive value of RDW for survival in young women with breast cancer. We reviewed 203 consecutive young female patients (under 40) with invasive breast cancer diagnosed at the First Affiliated Hospital of Wenzhou Medical University between January 2008 and December 2012. Preoperational RDW, clinicopathological information, and prognostic data were collected. RDW levels were divided into 2 groups: 161 patients with low RDW (≤13.75%) and 42 patients with high RDW (>13.75%). Clinicopathological differences between the 2 groups were calculated by chi-squared test and Wilcoxon rank-sum test. Kaplan–Meier survival analysis and Cox proportional hazard regression analyses were used to examine the effect of RDW on survival. We found that high RDW was significantly associated with larger tumor size (P = 0.002), positive lymph node metastases (P = 0.011), and advanced stages (P = 0.004). Patients with high RDW showed significantly lower disease-free survival (DFS; P < 0.001) and lower overall survival (OS) rate (P < 0.001) than patients with low RDW. Moreover, the Cox regression multivariate analysis revealed that high pretreatment DRW was independently correlated with poor DFS and OS, with hazard ratio 4.819 (95% confidence interval [CI] 2.291–10.138, P < 0.001) and 5.887 (95% CI 1.666–20.802, P = 0.006), respectively. In conclusion, our study demonstrated that pretreatment RDW may be associated with DFS and OS in young women with breast cancer. Further validation and feasibility studies are required before the result of our study can be considered for clinical practice. PMID:27124030

  3. [Yes to research, no to utilization? Medical, pharmacological and toxicological utilization of human embryonic stem cells from an ethical point of view].

    PubMed

    Kress, H

    2008-09-01

    In exceptional cases, the German Stem Cell Act allows research on human embryonic stem cells. However, it does not allow the implementation of the research results if this in turn requires the use of further embryonic stem cell lines. It has, in the meantime, transpired that such research results could be of concrete use. Thus, in the distant future, it could be used in the clinical treatment of patients. Already in the nearer future the use of human embryonic stem cell lines can be envisaged for both the development and testing of medicines as well as in the field of toxicology. To this end, research concerning embryo toxicity and neurotoxicity is ground-breaking. The toxicological and pharmacological use of human embryonic stem cell lines should serve the protection of human health as well as the safe and reliable use of medicines. In addition, animal experiments could be reduced, which is desirable from a point of view of animal protection ethics. Since research on human embryonic stem cell lines is actually permitted in Germany, the use of the respective research results should be allowed all the more. This follows from the basic human right to health protection and health care. Legal ambiguities, which still exist in this respect, should be removed.

  4. BdCESA7, BdCESA8, and BdPMT utility promoter constructs for targeted expression to secondary cell-wall-forming cells of grasses

    DOE PAGESBeta

    Petrik, Deborah L.; Cass, Cynthia L.; Padmakshan, Dharshana; Foster, Cliff E.; Vogel, John P.; Karlen, Steven D.; Ralph, John; Sedbrook, John C.

    2016-02-04

    Utility vectors with promoters that confer desired spatial and temporal expression patterns are useful tools for studying gene and cellular function and for industrial applications. To target the expression of DNA sequences of interest to cells forming plant secondary cell walls, which generate most of the vegetative biomass, upstream regulatory sequences of the Brachypodium distachyon lignin biosynthetic gene BdPMT and the cellulose synthase genes BdCESA7 and BdCESA8 were isolated and cloned into binary vectors designed for Agrobacterium-mediated transformation of monocots. Expression patterns were assessed using the β-glucuronidase gene GUSPlus and X-glucuronide staining. All three promoters showed strong expression levels inmore » stem tissue at the base of internodes where cell wall deposition is most active, in both vascular bundle xylem vessels and tracheids, and in interfascicular tissues, with expression less pronounced in developmentally older tissues. In leaves, BdCESA7 and BdCESA8 promoter-driven expression was strongest in leaf veins, leaf margins, and trichomes; relatively weaker and patchy expression was observed in the epidermis. BdPMT promoter-driven expression was similar to the BdCESA promoters expression patterns, including strong expression in trichomes. The intensity and extent of GUS staining varied considerably between transgenic lines, suggesting that positional effects influenced promoter activity. Introducing the BdPMT and BdCESA8 Open Reading Frames into BdPMT and BdCESA8 utility promoter binary vectors, respectively, and transforming those constructs into Brachypodium pmt and cesa8 loss-of-function mutants resulted in rescue of the corresponding mutant phenotypes. This work therefore validates the functionality of these utility promoter binary vectors for use in Brachypodium and likely other grass species. Lastly, the identification, in Bdcesa8-1 T-DNA mutant stems, of an 80% reduction in crystalline cellulose levels confirms that the

  5. High-Resolution Microfluidic Single-Cell Transcriptional Profiling Reveals Clinically Relevant Subtypes among Human Stem Cell Populations Commonly Utilized in Cell-Based Therapies.

    PubMed

    Rennert, Robert C; Schäfer, Richard; Bliss, Tonya; Januszyk, Michael; Sorkin, Michael; Achrol, Achal S; Rodrigues, Melanie; Maan, Zeshaan N; Kluba, Torsten; Steinberg, Gary K; Gurtner, Geoffrey C

    2016-01-01

    Stem cell therapies can promote neural repair and regeneration, yet controversy regarding optimal cell source and mechanism of action has slowed clinical translation, potentially due to undefined cellular heterogeneity. Single-cell resolution is needed to identify clinically relevant subpopulations with the highest therapeutic relevance. We combine single-cell microfluidic analysis with advanced computational modeling to study for the first time two common sources for cell-based therapies, human NSCs and MSCs. This methodology has the potential to logically inform cell source decisions for any clinical application. PMID:27047447

  6. Highly efficient colorimetric detection of target cancer cells utilizing superior catalytic activity of graphene oxide-magnetic-platinum nanohybrids.

    PubMed

    Kim, Moon Il; Kim, Min Su; Woo, Min-Ah; Ye, Youngjin; Kang, Kyoung Suk; Lee, Jinwoo; Park, Hyun Gyu

    2014-01-01

    Enzyme-linked immunosorbent assays (ELISAs) have most widely been applied in immunoassays for several decades. However, several unavoidable limitations (e.g., instability caused by structural unfolding) of natural enzymes have hindered their widespread applications. Here, we describe a new nanohybrid consisting of Fe₃O₄ magnetic nanoparticles (MNPs) and platinum nanoparticles (Pt NPs), simultaneously immobilized on the surface of graphene oxide (GO). By synergistically integrating highly catalytically active Pt NPs and MNPs on GO whose frameworks possess high substrate affinity, the nanohybrid is able to achieve up to a 30-fold higher maximal reaction velocity (V(max)) compared to that of free GO for the colorimetric reaction of the peroxidase substrate, 3,3',5,5'-tetramethylbenzidine (TMB), and enable rapid detection of target cancer cells. Specifically, using this new assay system, clinically important breast cancer cells are detected in a 5 min time period at room temperature with high specificity and sensitivity. The remarkably high capability to catalyze oxidation reactions could allow the nanohybrid to replace conventional peroxidase-based immunoassay systems as part of new, rapid, robust and convenient assay systems which can be widely utilized for the identification of important target molecules.

  7. Combined energy production and waste management in manned spacecraft utilizing on-demand hydrogen production and fuel cells

    NASA Astrophysics Data System (ADS)

    Elitzur, Shani; Rosenband, Valery; Gany, Alon

    2016-11-01

    Energy supply and waste management are among the most significant challenges in human spacecraft. Great efforts are invested in managing solid waste, recycling grey water and urine, cleaning the atmosphere, removing CO2, generating and saving energy, and making further use of components and products. This paper describes and investigates a concept for managing waste water and urine to simultaneously produce electric and heat energies as well as fresh water. It utilizes an original technique for aluminum activation to react spontaneously with water at room temperature to produce hydrogen on-site and on-demand. This reaction has further been proven to be effective also when using waste water and urine. Applying the hydrogen produced in a fuel cell, one obtains electric energy as well as fresh (drinking) water. The method was compared to the traditional energy production technology of the Space Shuttle, which is based on storing the fuel cell reactants, hydrogen and oxygen, in cryogenic tanks. It is shown that the alternative concept presented here may provide improved safety, compactness (reduction of more than one half of the volume of the hydrogen storage system), and management of waste liquids for energy generation and drinking water production. Nevertheless, it adds mass compared to the cryogenic hydrogen technology. It is concluded that the proposed method may be used as an emergency and backup power system as well as an additional hydrogen source for extended missions in human spacecraft.

  8. Immunocytochemistry and Image Analysis of Beta-Catenin Redistribution in Normal Human Colon Cell Cultures Treated with Disinfection By-Products.

    EPA Science Inventory

    Epidemiological studies have shown an association between the consumption of chlorinated drinking water and increased risk for colon cancer. In vivo studies proved that rodents exposed to chlorination disinfection byproducts (DBPs) developed aberrant crypt foci (ACF) in t...

  9. Utilization of modified surfactant-associated protein B for delivery of DNA to airway cells in culture.

    PubMed Central

    Baatz, J E; Bruno, M D; Ciraolo, P J; Glasser, S W; Stripp, B R; Smyth, K L; Korfhagen, T R

    1994-01-01

    Pulmonary surfactant lines the airway epithelium and creates a potential barrier to successful transfection of the epithelium in vivo. Based on the functional properties of pulmonary surfactant protein B (SP-B) and the fact that this protein is neither toxic nor immunogenic in the airway, we hypothesized that SP-B could be modified to deliver DNA to airway cells. We have modified native bovine SP-B by the covalent linkage of poly(lysine) (average molecular mass of 3.3 or 10 kDa) to the N terminus of SP-B and formed complexes between a test plasmid and the modified SP-B. Transfection efficiency was determined by transfection of pulmonary adenocarcinoma cells (H441) in culture with the test plasmid pCPA-RSV followed by measurement of activity of the reporter gene encoding chloramphenicol acetyltransferase (CAT). Transfections were performed with DNA.protein complexes using poly(lysine)10kDa-SP-B ([Lys]10kDa-SP-B) or poly(lysine)3.3kDa-SP-B ([Lys]3.3kDa-SP-B), and results were compared with transfections using unmodified poly(lysine).DNA, unmodified SP-B.DNA, or DNA only. For [Lys]10kDa-SP-B.pCPA-RSV preparations, CAT activity was readily detectable above the background of [Lys]3.3kDa-SP-B or unmodified SP-B. The SP-B-poly(lysine) conjugates were effective over a broad range of protein-to-DNA molar ratios, although they were optimal at approximately 500:1-1000:1. Transfection efficiency varied with the tested cell line but was not specific to airway cells. Addition of replication-defective adenovirus to the [Lys]10kDa-SP-B.pCPA-RSV complex enhanced CAT activity about 30-fold with respect to that produced by the [Lys]10kDa-SP-B.pCPA-RSV complex alone. This increase suggests routing of the adenoviral.[Lys]10kDa-SP-B.pCPA-RSV complex through an endosomal pathway. Effects of covalent modification on the secondary structure of SP-B were examined by Fourier transform infrared spectrometry (FTIR). Results of FTIR indicated that the conformation of [Lys]10kDa-SP-B was

  10. Utilization of a photoactivatable antigen system to examine B-cell probing termination and the B-cell receptor sorting mechanisms during B-cell activation

    PubMed Central

    Wang, Jing; Tang, Shan; Wan, Zhengpeng; Gao, Yiren; Cao, Yiyun; Yi, Junyang; Si, Yanyan; Zhang, Haowen; Liu, Lei; Liu, Wanli

    2016-01-01

    Antigen binding to the B-cell receptor (BCR) induces several responses, resulting in B-cell activation, proliferation, and differentiation. However, it has been difficult to study these responses due to their dynamic, fast, and transient nature. Here, we attempted to solve this problem by developing a controllable trigger point for BCR and antigen recognition through the construction of a photoactivatable antigen, caged 4-hydroxy-3-nitrophenyl acetyl (caged-NP). This photoactivatable antigen system in combination with live cell and single molecule imaging techniques enabled us to illuminate the previously unidentified B-cell probing termination behaviors and the precise BCR sorting mechanisms during B-cell activation. B cells in contact with caged-NP exhibited probing behaviors as defined by the unceasing extension of membrane pseudopods in random directions. Further analyses showed that such probing behaviors are cell intrinsic with strict dependence on F-actin remodeling but not on tonic BCR signaling. B-cell probing behaviors were terminated within 4 s after photoactivation, suggesting that this response was sensitive and specific to BCR engagement. The termination of B-cell probing was concomitant with the accumulation response of the BCRs into the BCR microclusters. We also determined the Brownian diffusion coefficient of BCRs from the same B cells before and after BCR engagement. The analysis of temporally segregated single molecule images of both BCR and major histocompatibility complex class I (MHC-I) demonstrated that antigen binding induced trapping of BCRs into the BCR microclusters is a fundamental mechanism for B cells to acquire antigens. PMID:26764382

  11. Biological Effects of Particles with Very High Energy Deposition on Mammalian Cells Utilizing the Brookhaven Tandem Van de Graaff Accelerator

    NASA Technical Reports Server (NTRS)

    Saha, Janapriya; Cucinotta, Francis A.; Wang, Minli

    2013-01-01

    High LET radiation from GCR (Galactic Cosmic Rays) consisting mainly of high charge and energy (HZE) nuclei and secondary protons and neutrons, and secondaries from protons in SPE (Solar Particle Event) pose a major health risk to astronauts due to induction of DNA damage and oxidative stress. Experiments with high energy particles mimicking the space environment for estimation of radiation risk are being performed at NASA Space Radiation Laboratory at BNL. Experiments with low energy particles comparing to high energy particles of similar LET are of interest for investigation of the role of track structure on biological effects. For this purpose, we report results utilizing the Tandem Van de Graaff accelerator at BNL. The primary objective of our studies is to elucidate the influence of high vs low energy deposition on track structure, delta ray contribution and resulting biological responses. These low energy ions are of special relevance as these energies may occur following absorption through the spacecraft and shielding materials in human tissues and nuclear fragments produced in tissues by high energy protons and neutrons. This study will help to verify the efficiency of these low energy particles and better understand how various cell types respond to them.

  12. Cytometric comparisons between circulating tumor cells from prostate cancer patients and the prostate tumor derived LNCaP cell line

    PubMed Central

    Lazar, Daniel C.; Cho, Edward H.; Luttgen, Madelyn S.; Metzner, Thomas J.; Uson, Maria Loressa; Torrey, Melissa; Gross, Mitchell E.; Kuhn, Peter

    2012-01-01

    Many important experiments in cancer research are initiated with cell line data analysis due to the ease of accessibility and utilization. Recently, the ability to capture and characterize circulating tumor cells (CTCs) has become more prevalent in the research setting. This ability to detect, isolate, and analyze CTCs allows us to directly compare specific protein expression levels found in patient CTCs to cell lines. In this study, we use immunocytochemistry to compare the protein expression levels of total cytokeratin (CK) and androgen receptor (AR) in CTCs and cell lines from patients with prostate cancer to determine what translational insights might be gained through the use of cell line data. A non-enrichment CTC detection assay enables us to compare cytometric features and relative expression levels of CK and AR by indirect immunofluorescence from prostate cancer patients against the prostate cancer cell line LNCaP. We measured physical characteristics of these two groups and observed significant differences in cell size, fluorescence intensity, and nuclear to cytoplasmic (N/C) ratio. We hope that these experiments will initiate a foundation to allow cell line data to be compared against characteristics of primary cells from patients. PMID:22306736

  13. Application of a novel population of multipotent stem cells derived from skin fibroblasts as donor cells in bovine SCNT.

    PubMed

    Pan, Shaohui; Chen, Wuju; Liu, Xu; Xiao, Jiajia; Wang, Yanqin; Liu, Jun; Du, Yue; Wang, Yongsheng; Zhang, Yong

    2015-01-01

    Undifferentiated stem cells are better donor cells for somatic cell nuclear transfer (SCNT), resulting in more offspring than more differentiated cells. While various stem cell populations have been confirmed to exist in the skin, progress has been restricted due to the lack of a suitable marker for their prospective isolation. To address this fundamental issue, a marker is required that could unambiguously prove the differentiation state of the donor cells. We therefore utilized magnetic activated cell sorting (MACS) to separate a homogeneous population of small SSEA-4(+) cells from a heterogeneous population of bovine embryonic skin fibroblasts (BEF). SSEA-4(+) cells were 8-10 μm in diameter and positive for alkaline phosphatase (AP). The percentage of SSEA-4(+) cells within the cultured BEF population was low (2-3%). Immunocytochemistry and PCR analyses revealed that SSEA-4(+) cells expressed pluripotency-related markers, and could differentiate into cells comprising all three germ layers in vitro. They remained undifferentiated over 20 passages in suspension culture. In addition, cloned embryos derived from SSEA-4 cells showed significant differences in cleavage rate and blastocyst development when compared with those from BEF and SSEA-4(-) cells. Moreover, blastocysts derived from SSEA-4(+) cells showed a higher total cell number and lower apoptotic index as compared to BEF and SSEA-4(-) derived cells. It is well known that nuclei from pluripotent stem cells yield a higher cloning efficiency than those from adult somatic cells, however, pluripotent stem cells are relatively difficult to obtain from bovine. The SSEA-4(+) cells described in the current study provide an attractive candidate for SCNT and a promising platform for the generation of transgenic cattle.

  14. Utility of Cell-Block of Bronchial Washings in Diagnosis of Lung Cancer- A Comparative Analysis with Conventional Smear Cytology

    PubMed Central

    Vadala, Rohit; Mandrekar, Suresh

    2016-01-01

    Introduction Bronchoscopy is a safe & effective means of diagnosing bronchogenic carcinoma with a varying diagnostic yield of different bronchoscopic procedures. Cell-Block (CB) preparation of cytology specimen has been shown to increase the diagnostic yield further. To the authors’ knowledge, the diagnostic value of CB as an adjunct to conventional smear cytology (CS) of bronchial washing specimens in the detection of bronchogenic carcinoma has not been well evaluated. Aim The present study was aimed to evaluate the diagnostic utility of CB of bronchial washings when compared with CS. Materials and Methods A total of 104 patients of suspected bronchogenic carcinoma were subjected to bronchoscopy as per British Thoracic Society (BTS) protocol. Bronchial biopsy, brushings and washings were collected. Smears were prepared immediately of bronchial washings and another aliquot was subjected to CB preparation and further processing by paraffin embedding and H&E staining. Results Out of 104 patients, 92 were diagnosed by bronchoscopy with a cumulative diagnostic yield of all sampling techniques being 88.46%. Yield of CB of bronchial washings (44.23%) was higher than Bronchial washings – conventional smears (36.53%). CB detected additional 8 cases of malignancy where corresponding bronchial washings-conventional smears were negative. Exclusive diagnosis by CB was obtained in 2 cases. Brushings and biopsy confirmed malignancy in 49.03% and 57.69% cases. Conclusion CB of bronchial washings had a higher yield as compared to corresponding conventional smears. Increase in yield was also noted when CB of bronchial washings was combined with biopsy and compared to bronchial washings- conventional smears combined with biopsy. In limited resource settings, CB preparation is a simple method that increases diagnostic yield of flexible bronchoscopy, is cost effective & hence can be routinely used. The immunohistochemical and molecular studies are possible with CB only, which is a

  15. Prognostic Utility of Cell Cycle Progression Score in Men With Prostate Cancer After Primary External Beam Radiation Therapy

    SciTech Connect

    Freedland, Stephen J.; Gerber, Leah; Reid, Julia; Welbourn, William; Tikishvili, Eliso; Park, Jimmy; Younus, Adib; Gutin, Alexander; Sangale, Zaina; Lanchbury, Jerry S.; Salama, Joseph K.; Stone, Steven

    2013-08-01

    Purpose: To evaluate the prognostic utility of the cell cycle progression (CCP) score, a RNA signature based on the average expression level of 31 CCP genes, for predicting biochemical recurrence (BCR) in men with prostate cancer treated with external beam radiation therapy (EBRT) as their primary curative therapy. Methods and Materials: The CCP score was derived retrospectively from diagnostic biopsy specimens of men diagnosed with prostate cancer from 1991 to 2006 (n=141). All patients were treated with definitive EBRT; approximately half of the cohort was African American. Outcome was time from EBRT to BCR using the Phoenix definition. Median follow-up for patients without BCR was 4.8 years. Association with outcome was evaluated by Cox proportional hazards survival analysis and likelihood ratio tests. Results: Of 141 patients, 19 (13%) had BCR. The median CCP score for patient samples was 0.12. In univariable analysis, CCP score significantly predicted BCR (P=.0017). The hazard ratio for BCR was 2.55 for 1-unit increase in CCP score (equivalent to a doubling of gene expression). In a multivariable analysis that included Gleason score, prostate-specific antigen, percent positive cores, and androgen deprivation therapy, the hazard ratio for CCP changed only marginally and remained significant (P=.034), indicating that CCP provides prognostic information that is not provided by standard clinical parameters. With 10-year censoring, the CCP score was associated with prostate cancer-specific mortality (P=.013). There was no evidence for interaction between CCP and any clinical variable, including ethnicity. Conclusions: Among men treated with EBRT, the CCP score significantly predicted outcome and provided greater prognostic information than was available with clinical parameters. If validated in a larger cohort, CCP score could identify high-risk men undergoing EBRT who may need more aggressive therapy.

  16. Pt skin on AuCu intermetallic substrate: a strategy to maximize Pt utilization for fuel cells.

    PubMed

    Wang, Gongwei; Huang, Bing; Xiao, Li; Ren, Zhandong; Chen, Hao; Wang, Deli; Abruña, Héctor D; Lu, Juntao; Zhuang, Lin

    2014-07-01

    The dependence on Pt catalysts has been a major issue of proton-exchange membrane (PEM) fuel cells. Strategies to maximize the Pt utilization in catalysts include two main approaches: to put Pt atoms only at the catalyst surface and to further enhance the surface-specific catalytic activity (SA) of Pt. Thus far there has been no practical design that combines these two features into one single catalyst. Here we report a combined computational and experimental study on the design and implementation of Pt-skin catalysts with significantly improved SA toward the oxygen reduction reaction (ORR). Through screening, using density functional theory (DFT) calculations, a Pt-skin structure on AuCu(111) substrate, consisting of 1.5 monolayers of Pt, is found to have an appropriately weakened oxygen affinity, in comparison to that on Pt(111), which would be ideal for ORR catalysis. Such a structure is then realized by substituting the Cu atoms in three surface layers of AuCu intermetallic nanoparticles (AuCu iNPs) with Pt. The resulting Pt-skinned catalyst (denoted as Pt(S)AuCu iNPs) has been characterized in depth using synchrotron XRD, XPS, HRTEM, and HAADF-STEM/EDX, such that the Pt-skin structure is unambiguously identified. The thickness of the Pt skin was determined to be less than two atomic layers. Finally the catalytic activity of Pt(S)AuCu iNPs toward the ORR was measured via rotating disk electrode (RDE) voltammetry through which it was established that the SA was more than 2 times that of a commercial Pt/C catalyst. Taking into account the ultralow Pt loading in Pt(S)AuCu iNPs, the mass-specific catalytic activity (MA) was determined to be 0.56 A/mg(Pt)@0.9 V, a value that is well beyond the DOE 2017 target for ORR catalysts (0.44 A/mg(Pt)@0.9 V). These findings provide a strategic design and a realizable approach to high-performance and Pt-efficient catalysts for fuel cells.

  17. Fabrication of solid state dye sensitized solar cells utilizing vapor phase polymerized poly(3,4-ethylenedioxythiophene) hole conducting layer

    NASA Astrophysics Data System (ADS)

    Skorenko, Kenneth H.

    There is a need for sustainable and renewable energy sources that can be used in both grid and off-grid structured systems. Photovoltaic devices have been used to generate electrical energy by capturing and converting photons from the sun. Dye sensitized solar cells (DSSC) have gained attention due to their consistent energy generation during indirect sunlight. Furthermore, DSSC can be applied as a flexible device and gain benefits from the low cost roll to roll manufacturing. With this in mind, we have taken steps toward optimizing a DSSC device for use as a solid state solar cell using conducting polymers. Typically DSSC use a liquid electrolyte as a hole conducting layer used to direct the separation of electron -- hole pairs. This liquid electrolyte comes with problems that can be subverted using conducting polymers. Poly(3,4 -- ethylenedioxythiophene) (PEDOT), is a conducting thiophene that is tailored to have enhanced conductivity. We show that a vapor phase polymerization (VPP) of PEDOT can be used as a hole conducting layer in a solid state DSSC device. To this end we have investigated the electrical properties of the VPP PEDOT films in order to understand how the morphology and conductive domains relate to a polymers conductivity. Using 4 point probe we have measure the sheet resistance of the film, as well as how the films resistance is altered during stress tests. Scanning electron microscopy has been utilized to compare morphologies of different PEDOT films and see how surface morphology impacts the conductance measured. Using conductive atomic force microscopy we can look at the conductive domains between VPP PEDOT and PEDOT:PSS films. We saw that conductive domains of the VPP PEDOT are not only more conductive but also much larger in size and widespread throughout the film. We show that there is formation of PEDOT through optical spectroscopy and structural characterization such as UV/Vis and Raman spectroscopy as well as X-ray diffraction. When

  18. Giving cell phones to pregnant women and improving services may increase primary health facility utilization: a case–control study of a Nigerian project

    PubMed Central

    2014-01-01

    Background Worldwide, about 287 000 women die each year from mostly preventable complications related to pregnancy and childbirth. A disproportionately high number of these deaths occur in sub-Saharan Africa. The Abiye (‘Safe Motherhood’) project in the Ifedore Local Government Area (LGA) of Ondo-State of Nigeria aimed at improving facility utilization and maternal health through the use of cell phones and generally improved health care services for pregnant women, including Health Rangers, renovated Health Centres, and improved means of transportation. Methods A one-year sample of retrospective data was collected from hospital records and patients’ case files from Ifedore (the project area) and Idanre (control area) and was analyzed to determine healthcare facility utilization rates in each location. Semi-structured questionnaires were used to generate supplemental data. Results The total facility utilization rate of pregnant women was significantly higher in Ifedore than in Idanre. The facility utilization rate of the primary health care centres was significantly higher in Ifedore than in Idanre. The number of recorded cases of the five major causes of maternal death in the two LGAs was not significantly different, possibly because the project was new. Conclusions Giving cell phones to pregnant women and generally improving services could increase their utilization of the primary healthcare system. PMID:24438150

  19. Labview utilities

    2011-09-30

    The software package provides several utilities written in LabView. These utilities don't form independent programs, but rather can be used as a library or controls in other labview programs. The utilities include several new controls (xcontrols), VIs for input and output routines, as well as other 'helper'-functions not provided in the standard LabView environment.

  20. Utilization of a fuel cell power plant for the capture and conversion of gob well gas. Final report, June--December, 1995

    SciTech Connect

    Przybylic, A.R.; Haynes, C.D.; Haskew, T.A.; Boyer, C.M. II; Lasseter, E.L.

    1995-12-01

    A preliminary study has been made to determine if a 200 kW fuel cell power plant operating on variable quality coalbed methane can be placed and successfully operated at the Jim Walter Resources No. 4 mine located in Tuscaloosa County, Alabama. The purpose of the demonstration is to investigate the effects of variable quality (50 to 98% methane) gob gas on the output and efficiency of the power plant. To date, very little detail has been provided concerning the operation of fuel cells in this environment. The fuel cell power plant will be located adjacent to the No. 4 mine thermal drying facility rated at 152 M British thermal units per hour. The dryer burns fuel at a rate of 75,000 cubic feet per day of methane and 132 tons per day of powdered coal. The fuel cell power plant will provide 700,000 British thermal units per hour of waste heat that can be utilized directly in the dryer, offsetting coal utilization by approximately 0.66 tons per day and providing an avoided cost of approximately $20 per day. The 200 kilowatt electrical power output of the unit will provide a utility cost reduction of approximately $3,296 each month. The demonstration will be completely instrumented and monitored in terms of gas input and quality, electrical power output, and British thermal unit output. Additionally, real-time power pricing schedules will be applied to optimize cost savings. 28 refs., 35 figs., 13 tabs.

  1. p53 directly regulates the transcription of the human frataxin gene and its lack of regulation in tumor cells decreases the utilization of mitochondrial iron.

    PubMed

    Shimizu, Rina; Lan, Nguyen Ngoc; Tai, Tran Tien; Adachi, Yuka; Kawazoe, Asako; Mu, Anfeng; Taketani, Shigeru

    2014-11-01

    Mitochondrial frataxin functions in iron homeostasis, biogenesis of iron-sulfur clusters, protection from oxidative stress and apoptosis, and as a tumor suppressor protein. We examined regulation of the expression of the human frataxin by p53. Pifithrin-α, an inhibitor of p53 function, and knockdown of p53 decreased the level of frataxin mRNA in human kidney HEK 293T cells. The transcriptional activity of the human frataxin gene is enhanced by the proximal promoter containing the p53-responsive element (p53RE) on the gene. Chromatin immunoprecipitation assay and electrophoretic mobility shift assay confirmed the binding of p53 to the human frataxin p53RE. The expression of wild-type p53 in human cancer HeLa cells increased the reporter activity carrying p53RE at the region of -209 to -200bp of the frataxin promoter. Finally, when the HeLa cells overexpressing frataxin were treated with 5-aminolevulinic acid (ALA), there was less accumulation of protoporphyrin than HeLa control cells, and it was sharply decreased by the addition of iron citrate, suggesting that the utilization of mitochondrial iron for heme biosynthesis can be dependent on the level of frataxin. Alternatively, the low expression of frataxin not regulated by p53 in tumor cells lowers the utilization of iron in mitochondria, causing the tumor-specific ALA-induced accumulation of protoporphyrin.

  2. Transcriptome Sequencing (RNAseq) Enables Utilization of Formalin-Fixed, Paraffin-Embedded Biopsies with Clear Cell Renal Cell Carcinoma for Exploration of Disease Biology and Biomarker Development.

    PubMed

    Eikrem, Oystein; Beisland, Christian; Hjelle, Karin; Flatberg, Arnar; Scherer, Andreas; Landolt, Lea; Skogstrand, Trude; Leh, Sabine; Beisvag, Vidar; Marti, Hans-Peter

    2016-01-01

    Formalin-fixed, paraffin-embedded (FFPE) tissues are an underused resource for molecular analyses. This proof of concept study aimed to compare RNAseq results from FFPE biopsies with the corresponding RNAlater® (Qiagen, Germany) stored samples from clear cell renal cell carcinoma (ccRCC) patients to investigate feasibility of RNAseq in archival tissue. From each of 16 patients undergoing partial or full nephrectomy, four core biopsies, such as two specimens with ccRCC and two specimens of adjacent normal tissue, were obtained with a 16g needle. One normal and one ccRCC tissue specimen per patient was stored either in FFPE or RNAlater®. RNA sequencing libraries were generated applying the new Illumina TruSeq® Access library preparation protocol. Comparative analysis was done using voom/Limma R-package. The analysis of the FFPE and RNAlater® datasets yielded similar numbers of detected genes, differentially expressed transcripts and affected pathways. The FFPE and RNAlater datasets shared 80% (n = 1106) differentially expressed genes. The average expression and the log2 fold changes of these transcripts correlated with R2 = 0.97, and R2 = 0.96, respectively. Among transcripts with the highest fold changes in both datasets were carbonic anhydrase 9 (CA9), neuronal pentraxin-2 (NPTX2) and uromodulin (UMOD) that were confirmed by immunohistochemistry. IPA revealed the presence of gene signatures of cancer and nephrotoxicity, renal damage and immune response. To simulate the feasibility of clinical biomarker studies with FFPE samples, a classifier model was developed for the FFPE dataset: expression data for CA9 alone had an accuracy, specificity and sensitivity of 94%, respectively, and achieved similar performance in the RNAlater dataset. Transforming growth factor-ß1 (TGFB1)-regulated genes, epithelial to mesenchymal transition (EMT) and NOTCH signaling cascade may support novel therapeutic strategies. In conclusion, in this proof of concept study, RNAseq data

  3. Transcriptome Sequencing (RNAseq) Enables Utilization of Formalin-Fixed, Paraffin-Embedded Biopsies with Clear Cell Renal Cell Carcinoma for Exploration of Disease Biology and Biomarker Development

    PubMed Central

    Eikrem, Oystein; Beisland, Christian; Hjelle, Karin; Flatberg, Arnar; Scherer, Andreas; Landolt, Lea; Skogstrand, Trude; Leh, Sabine; Beisvag, Vidar; Marti, Hans-Peter

    2016-01-01

    Formalin-fixed, paraffin-embedded (FFPE) tissues are an underused resource for molecular analyses. This proof of concept study aimed to compare RNAseq results from FFPE biopsies with the corresponding RNAlater® (Qiagen, Germany) stored samples from clear cell renal cell carcinoma (ccRCC) patients to investigate feasibility of RNAseq in archival tissue. From each of 16 patients undergoing partial or full nephrectomy, four core biopsies, such as two specimens with ccRCC and two specimens of adjacent normal tissue, were obtained with a 16g needle. One normal and one ccRCC tissue specimen per patient was stored either in FFPE or RNAlater®. RNA sequencing libraries were generated applying the new Illumina TruSeq® Access library preparation protocol. Comparative analysis was done using voom/Limma R-package. The analysis of the FFPE and RNAlater® datasets yielded similar numbers of detected genes, differentially expressed transcripts and affected pathways. The FFPE and RNAlater datasets shared 80% (n = 1106) differentially expressed genes. The average expression and the log2 fold changes of these transcripts correlated with R2 = 0.97, and R2 = 0.96, respectively. Among transcripts with the highest fold changes in both datasets were carbonic anhydrase 9 (CA9), neuronal pentraxin-2 (NPTX2) and uromodulin (UMOD) that were confirmed by immunohistochemistry. IPA revealed the presence of gene signatures of cancer and nephrotoxicity, renal damage and immune response. To simulate the feasibility of clinical biomarker studies with FFPE samples, a classifier model was developed for the FFPE dataset: expression data for CA9 alone had an accuracy, specificity and sensitivity of 94%, respectively, and achieved similar performance in the RNAlater dataset. Transforming growth factor-ß1 (TGFB1)-regulated genes, epithelial to mesenchymal transition (EMT) and NOTCH signaling cascade may support novel therapeutic strategies. In conclusion, in this proof of concept study, RNAseq data

  4. Utility of Induced Pluripotent Stem Cells for the Study and Treatment of Genetic Diseases: Focus on Childhood Neurological Disorders

    PubMed Central

    Barral, Serena; Kurian, Manju A.

    2016-01-01

    The study of neurological disorders often presents with significant challenges due to the inaccessibility of human neuronal cells for further investigation. Advances in cellular reprogramming techniques, have however provided a new source of human cells for laboratory-based research. Patient-derived induced pluripotent stem cells (iPSCs) can now be robustly differentiated into specific neural subtypes, including dopaminergic, inhibitory GABAergic, motorneurons and cortical neurons. These neurons can then be utilized for in vitro studies to elucidate molecular causes underpinning neurological disease. Although human iPSC-derived neuronal models are increasingly regarded as a useful tool in cell biology, there are a number of limitations, including the relatively early, fetal stage of differentiated cells and the mainly two dimensional, simple nature of the in vitro system. Furthermore, clonal variation is a well-described phenomenon in iPSC lines. In order to account for this, robust baseline data from multiple control lines is necessary to determine whether a particular gene defect leads to a specific cellular phenotype. Over the last few years patient-derived neural cells have proven very useful in addressing several mechanistic questions related to central nervous system diseases, including early-onset neurological disorders of childhood. Many studies report the clinical utility of human-derived neural cells for testing known drugs with repurposing potential, novel compounds and gene therapies, which then can be translated to clinical reality. iPSCs derived neural cells, therefore provide great promise and potential to gain insight into, and treat early-onset neurological disorders. PMID:27656126

  5. Transfer of two oligosaccharides to protein in a Chinese hamster ovary cell B211 which utilizes polyprenol for its N-linked glycosylation intermediates.

    PubMed

    Kaiden, A; Rosenwald, A G; Cacan, R; Verbert, A; Krag, S S

    1998-10-15

    B211, a glycosylation mutant isolated from Chinese hamster ovary cells, synthesizes 10- to 15-fold less Glc3Man9GlcNAc2-P-P-lipid, the substrate used by the oligosaccharide transferase in the synthesis of asparagine-linked glycoproteins. B211 cells are also 10- to 15-fold deficient in the glucosylation of oligosaccharide-lipid. Despite these properties, protein glycosylation in B211 cells proceeds at a level similar to (50% of) parental cells. We asked whether the near wild-type level of glycosylation was due to the transfer of alternative oligosaccharide structures to protein in B211 cells. The aberrant size of [35S]methionine-labeled VSV G protein and the increased percentage of endoglycosidase H-resistant tryptic peptides as compared to parental cells supported this hypothesis. B211 cells were labeled with [2-3H]mannose either for 1 min or for 1 h in the presence of glycoprotein-processing inhibitors so that the oligosaccharides initially transferred to protein could be analyzed. In addition to Glc3Man9GlcNAc2, a second, endoglycosidase H-resistant oligosaccharide was transferred whose structure was determined by alpha-mannosidase digestion, gel filtration chromatography, and HPLC to be Glc0,1Man5GlcNAc2. Finally, since the synthesis of reduced amounts of Glc3Man9GlcNAc2-P-P-lipid was also a phenotype seen in another glycosylation mutant, Lec9, we analyzed the long-chain prenol in B211 cells. B211 cells synthesized and utilized polyprenol rather than dolichol for all N-linked glycosylation intermediates as determined by HPLC analysis of [3H]mevalonate-labeled lipids. Cell fusions analyzed by similar techniques indicated that B211, originally isolated as a concanavalin A-resistant cell line, is in the Lec9 complementation group.

  6. An economic feasibility analysis of distributed electric power generation based upon the natural gas-fired fuel cell: a model of a central utility plant.

    SciTech Connect

    Not Available

    1993-06-30

    This central utilities plant model details the major elements of a central utilities plant for several classes of users. The model enables the analyst to select optional, cost effective, plant features that are appropriate to a fuel cell application. These features permit the future plant owner to exploit all of the energy produced by the fuel cell, thereby reducing the total cost of ownership. The model further affords the analyst an opportunity to identify avoided costs of the fuel cell-based power plant. This definition establishes the performance and capacity information, appropriate to the class of user, to support the capital cost model and the feasibility analysis. It is detailed only to the depth required to identify the major elements of a fuel cell-based system. The model permits the choice of system features that would be suitable for a large condominium complex or a residential institution such as a hotel, boarding school or prison. The user may also select large office buildings that are characterized by 12 to 16 hours per day of operation or industrial users with a steady demand for thermal and electrical energy around the clock.

  7. PLAG1 gene alterations in salivary gland pleomorphic adenoma and carcinoma ex-pleomorphic adenoma: a combined study using chromosome banding, in situ hybridization and immunocytochemistry.

    PubMed

    Martins, Carmo; Fonseca, Isabel; Roque, Lúcia; Pereira, Teresa; Ribeiro, Catarina; Bullerdiek, Jörn; Soares, Jorge

    2005-08-01

    Pleomorphic adenoma is the most common benign tumor of the salivary glands. It has marked histological diversity with epithelial, myoepithelial and mesenchymal-type cells arranged in a variety of architectural and differentiation patterns. Pleomorphic adenoma gene 1 (PLAG1), shown to be consistently rearranged in pleomorphic adenomas, is activated by chromosomal translocations involving 8q12, the chromosome region that is most frequently affected in these tumors. In this study, we evaluated PLAG1 involvement in salivary gland tumorigenesis by determining the frequency of its alterations in a selected group of 20 salivary gland tumors: 16 pleomorphic adenomas and four carcinomas ex-pleomorphic adenoma, having in common the presence of karyotypic chromosome 8 deviations, either structural, with 8q12 rearrangements, or numerical, with gain of chromosome 8. PLAG1 status was analyzed using in situ hybridization techniques, on metaphase cells, by fluorescence detection and/or interphase cells in paraffin sections, by chromogenic detection. Except for one pleomorphic adenoma case (5%) that lacked PLAG1 involvement, 17 tumors (85%), (14 pleomorphic adenomas and three carcinomas ex-pleomorphic adenoma) showed intragenic rearrangements of PLAG1 and the remaining two cases (10%), (one pleomorphic adenoma and one carcinoma ex-pleomorphic adenoma), had chromosome trisomy 8 only. To further investigate the role of PLAG1 on pleomorphic adenomas tumorigenesis, as well as the putative morphogenesis mechanism, we attempted to identify the cell types (epithelial vs myoepithelial) carrying 8q12/PLAG1 abnormalities by a combined phenotypic/genotypic analysis in four cases (three pleomorphic adenoma and one carcinoma ex-pleomorphic adenoma) characterized by 8q12 translocations and PLAG1 rearrangement. In these cases, both cells populations carried PLAG1 rearrangements. This finding further supports the pluripotent single-cell theory, which postulates that the tumor-initiated, modified

  8. Comparative Analysis of UV Irradiation Effects on Escherichia coli and Pseudomonas aeruginosa Bacterial Cells Utilizing Biological and Computational Approaches.

    PubMed

    Margaryan, A; Badalyan, H; Trchounian, A

    2016-09-01

    Microorganisms have a large number of tools to withstand different, and sometimes strong, environmental stresses, including irradiation, but this ability should be further evaluated for certain applications. Growth inhibition and morphological alterations of Escherichia coli M-17 and Pseudomonas aeruginosa GRP3 wild-type cells caused by UV-A irradiation have been detected in the present study. Comparative analysis was carried out using well-established microbiological methods (determination of specific growth rate, growth lag phase duration, and colony-forming unit number-CFU) and computational approaches, employing light microscopy and digital image analysis to evaluate bacterial cell morphology. Decreases in the specific growth rate, prolonged lag-phases, and lowered CFUs were observed after 5 and 10 min of UV irradiation (approx. 40 Gy) compared to the control (nonirradiated) cells. Accordingly, two computational parameters-the average bacterial cell surface area and the bacterial cell perimeter (i.e., of the 2D projection of bacterial cells in microscopy image)-were reduced. The ratio of bacterial cell surface area (S) to the square of the perimeter (p (2) ) was reduced after 5 min of irradiation, but after 10 min of irradiation the studied bacterial cells became flat cylinders. The revealed findings are concluded to be highly useful in developing new, rapid analysis methods to monitor environmental and UV irradiation effects on bacteria and to detect bacterial cell morphology alterations. PMID:27334536

  9. Method and apparatus for fabricating a thin-film solar cell utilizing a hot wire chemical vapor deposition technique

    DOEpatents

    Wang, Qi; Iwaniczko, Eugene

    2006-10-17

    A thin-film solar cell is provided. The thin-film solar cell comprises an a-SiGe:H (1.6 eV) n-i-p solar cell having a deposition rate of at least ten (10) .ANG./second for the a-SiGe:H intrinsic layer by hot wire chemical vapor deposition. A method for fabricating a thin film solar cell is also provided. The method comprises depositing a n-i-p layer at a deposition rate of at least ten (10) .ANG./second for the a-SiGe:H intrinsic layer.

  10. Nonphotochemical Hole-Burning Imaging Studies of in vitro Carcinoma and Normal Cells Utilizing a Mitochondrial Specific Dye

    SciTech Connect

    Richard Joseph Walsh

    2002-06-27

    Low temperature Nonphotochemical Hole Burning (NPHB) Spectroscopy of the dye rhodamine 800 (MF680) was applied for the purpose of discerning differences between cultured normal and carcinoma ovarian surface epithelial (OSE) cells. Both the cell lines were developed and characterized at the Mayo Clinic (Rochester, MN), with the normal cell line having been transfected with a strain of temperature sensitive Simian Virus 40 Large T Antigen (SV40) for the purpose of extending the life of the cell culture without inducing permanent changes in the characteristics of the cell line. The cationic lipophilic fluorophore rhodamine 800 preferentially locates in in situ mitochondria due to the high lipid composition of mitochondria and the generation of a large negative membrane potential (relative to the cellular cytoplasm) for oxidative phosphorylation. Results presented for NPHB of MF680 located in the cells show significant differences between the two cell lines. The results are interpreted on the basis of the NPHB mechanism and characteristic interactions between the host (cellular mitochondrial) and the guest (MF680) in the burning of spectral holes, thus providing an image of the cellular ultrastructure. Hole growth kinetics (HGK) were found to differ markedly between the two cell lines, with the carcinoma cell line burning at a faster average rate for the same exposure fluence. Theoretical fits to the data suggest a lower degree of structural heterogeneity in the carcinoma cell line relative to the normal cell line. Measurement of changes in the permanent dipole moment (f{Delta}{mu}) were accomplished by measurement of changes in hole width in response to the application of an external electric field (the Stark effect), and found that {Delta}{mu} values for the carcinoma line were 1.5x greater than those of the SV40 antigen-free normal analogs. These findings are interpreted in terms of effects from the mitochondrial membrane potential. Results for HGK on the scale of

  11. Nonphotochemical Hole-Burning Imaging Studies of In Vitro Carcinoma and Normal Cells Utilizing a Mitochondrial Specific Dye

    SciTech Connect

    Richard Joseph Walsh

    2002-08-01

    Low temperature Nonphotochemical Hole Burning (NPHB) Spectroscopy of the dye rhodamine 800 (MF680) was applied for the purpose of discerning differences between cultured normal and carcinoma ovarian surface epithelial (OSE) cells. Both the cell lines were developed and characterized at the Mayo Clinic (Rochester, MN), with the normal cell line having been transfected with a strain of temperature sensitive Simian Virus 40 Large T Antigen (SV40) for the purpose of extending the life of the cell culture without inducing permanent changes in the characteristics of the cell line. The cationic lipophilic fluorophore rhodamine 800 preferentially locates in in situ mitochondria due to the high lipid composition of mitochondria and the generation of a large negative membrane potential (relative to the cellular cytoplasm) for oxidative phosphorylation. Results presented for NPHB of MF680 located in the cells show significant differences between the two cell lines. The results are interpreted on the basis of the NPHB mechanism and characteristic interactions between the host (cellular mitochondrial) and the guest (MF680) in the burning of spectral holes, thus providing an image of the cellular ultrastructure. Hole growth kinetics (HGK) were found to differ markedly between the two cell lines, with the carcinoma cell line burning at a faster average rate for the same exposure fluence. Theoretical fits to the data suggest a lower degree of structural heterogeneity in the carcinoma cell line relative to the normal cell line. Measurement of changes in the permanent dipole moment (f{Delta}{mu})were accomplished by measurement of changes in hole width in response to the application of an external electric field (the Stark effect), and found that {Delta}{mu} values for the carcinoma line were 1.5x greater than those of the SV40 antigen-free normal analogs. These findings are interpreted in terms of effects from the mitochondrial membrane potential. Results for HGK on the scale of

  12. The Prospective Collection, Storage and Reporting of Data on Patients Undergoing Hematopoietic Stem Cell Transplantation Utilizing a Standard Preparative Regimen

    ClinicalTrials.gov

    2016-07-26

    Acute Myelogenous Leukemia; Acute Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Non-Hodgkin's Lymphoma; Hodgkin's Disease; Multiple Myeloma; Germ Cell Neoplasms; Myelodysplastic Syndromes; Chronic Lymphocytic Leukemia; Immunodeficiency Diseases

  13. Differential utilization of ketone bodies by neurons and glioma cell lines: a rationale for ketogenic diet as experimental glioma therapy

    PubMed Central

    2011-01-01

    Background Even in the presence of oxygen, malignant cells often highly depend on glycolysis for energy generation, a phenomenon known as the Warburg effect. One strategy targeting this metabolic phenotype is glucose restriction by administration of a high-fat, low-carbohydrate (ketogenic) diet. Under these conditions, ketone bodies are generated serving as an important energy source at least for non-transformed cells. Methods To investigate whether a ketogenic diet might selectively impair energy metabolism in tumor cells, we characterized in vitro effects of the principle ketone body 3-hydroxybutyrate in rat hippocampal neurons and five glioma cell lines. In vivo, a non-calorie-restricted ketogenic diet was examined in an orthotopic xenograft glioma mouse model. Results The ketone body metabolizing enzymes 3-hydroxybutyrate dehydrogenase 1 and 2 (BDH1 and 2), 3-oxoacid-CoA transferase 1 (OXCT1) and acetyl-CoA acetyltransferase 1 (ACAT1) were expressed at the mRNA and protein level in all glioma cell lines. However, no activation of the hypoxia-inducible factor-1α (HIF-1α) pathway was observed in glioma cells, consistent with the absence of substantial 3-hydroxybutyrate metabolism and subsequent accumulation of succinate. Further, 3-hydroxybutyrate rescued hippocampal neurons from glucose withdrawal-induced cell death but did not protect glioma cell lines. In hypoxia, mRNA expression of OXCT1, ACAT1, BDH1 and 2 was downregulated. In vivo, the ketogenic diet led to a robust increase of blood 3-hydroxybutyrate, but did not alter blood glucose levels or improve survival. Conclusion In summary, glioma cells are incapable of compensating for glucose restriction by metabolizing ketone bodies in vitro, suggesting a potential disadvantage of tumor cells compared to normal cells under a carbohydrate-restricted ketogenic diet. Further investigations are necessary to identify co-treatment modalities, e.g. glycolysis inhibitors or antiangiogenic agents that efficiently

  14. Bacterial Heat Shock Protein GroEL (Hsp64) Exerts Immunoregulatory Effects on T Cells by Utilizing Apoptosis

    PubMed Central

    Nalbant, Ayten; Kant, Melis

    2016-01-01

    Aggregatibacter actinomycetemcomitans (Aa) expresses a 64-kDa GroEL protein belonging to the heat shock family of proteins. This protein has been shown to influence human host cells, but the apoptotic capacity of the GroEL protein regarding T cells is not yet known. The purpose of this study was to investigate the ability of A. actinomycetemcomitans GroEL (AaGroEL) protein to induce human peripheral blood T-cell apoptosis. Endogenous, purified AaGroEL protein was used as an antigen. In AaGroEL-treated T cells, the data indicated that phosphatidylserine exposure, an early apoptotic event, was dose- and time-dependent. The AaGroEL-treated T cells were also positive for active caspase-3 in a dose-dependent manner. The rate of AaGroEL-induced apoptosis was suppressed by the addition of the general caspase inhibitor Z-VAD-FMK. Furthermore, cleaved caspase-8 bands (40/36 kDa and 23 kDa) were identified in cells responding to AaGroEL. DNA fragmentation was also detected in the AaGroEL-treated T cells. Overall, we demonstrated that the endogenous GroEL from A. actinomycetemcomitans has the capacity to induce T-cell apoptosis. PMID:27736933

  15. Membrane electrode assembly with enhanced platinum utilization for high temperature proton exchange membrane fuel cell prepared by catalyst coating membrane method

    NASA Astrophysics Data System (ADS)

    Liang, Huagen; Su, Huaneng; Pollet, Bruno G.; Linkov, Vladimir; Pasupathi, Sivakumar

    2014-11-01

    In this work, membrane electrode assemblies (MEAs) prepared by catalyst coating membrane (CCM) method are investigated for reduced platinum (Pt) loading and improved Pt utilization of high temperature proton exchange membrane fuel cell (PEMFC) based on phosphoric acid (PA)-doped poly(2,5-benzimidazole) (AB-PBI) membrane. The results show that CCM method exhibits significantly higher cell performance and Pt-specific power density than that of MEAs prepared with conventional gas diffusion electrode (GDE) under a low Pt loading level. In-suit cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) show that the MEAs prepared by the CCM method have a higher electrochemical surface area (ECSA), low cell ohmic resistance and low charge transfer resistance as compared to those prepared with GDEs at the same Pt loading.

  16. Safety, efficacy and utility of methods of transferring adhesive and cohesive Escherichia coli cells to microplates to avoid aerosols

    PubMed Central

    Ericksen, Bryan

    2015-01-01

    The virtual colony count (VCC) microbiological assay has been utilized for over a decade to measure the antimicrobial activity of peptides such as defensins and LL-37 against biosafety level (BSL)-1 and BSL-2 bacteria including Escherichia coli, Staphylococcus aureus, Bacillus cereus, and Enterobacter aerogenes.  In addition, a modified pipetting technique was presented in a 2011 study of defensin activity against the BSL-3 pathogen Bacillus anthracis.  Both studies were published in the journal Antimicrobial Agents and Chemotherapy.  Here I report that the method can also detect cross-contamination caused by aerosols utilizing the VCC method of data analysis by quantitative growth kinetics (QGK).  The QGK threshold time, or T t, equivalent to the cycle time C t reported in 1996 by Heid et al., precisely identifies when wells were inoculated. PMID:25671086

  17. Investigation of the effects of external current systems on the MAGSAT data utilizing grid cell modeling techniques

    NASA Technical Reports Server (NTRS)

    Klumpar, D. M. (Principal Investigator)

    1981-01-01

    Efforts devoted to reading MAGSAT data tapes in preparation for further analysis of the MAGSAT data are discussed. A modeling procedure developed to compute the magnetic fields at satellite orbit due to hypothesized current distributions in the ionosphere and magnetosphere is described. This technique utilizes a linear current element representation of the large-scale space-current system. Several examples of the model field perturbations computed along hypothetical satellite orbits are shown.

  18. Do circulating tumor cells, exosomes, and circulating tumor nucleic acids have clinical utility? A report of the association for molecular pathology.

    PubMed

    Gold, Bert; Cankovic, Milena; Furtado, Larissa V; Meier, Frederick; Gocke, Christopher D

    2015-05-01

    Diagnosing and screening for tumors through noninvasive means represent an important paradigm shift in precision medicine. In contrast to tissue biopsy, detection of circulating tumor cells (CTCs) and circulating tumor nucleic acids provides a minimally invasive method for predictive and prognostic marker detection. This allows early and serial assessment of metastatic disease, including follow-up during remission, characterization of treatment effects, and clonal evolution. Isolation and characterization of CTCs and circulating tumor DNA (ctDNA) are likely to improve cancer diagnosis, treatment, and minimal residual disease monitoring. However, more trials are required to validate the clinical utility of precise molecular markers for a variety of tumor types. This review focuses on the clinical utility of CTCs and ctDNA testing in patients with solid tumors, including somatic and epigenetic alterations that can be detected. A comparison of methods used to isolate and detect CTCs and some of the intricacies of the characterization of the ctDNA are also provided.

  19. Registration procedure for spatial correlation of physical energy deposition of particle irradiation and cellular response utilizing cell-fluorescent ion track hybrid detectors.

    PubMed

    Niklas, M; Zimmermann, F; Schlegel, J; Schwager, C; Debus, J; Jäkel, O; Abdollahi, A; Greilich, S

    2016-09-01

    The hybrid technology cell-fluorescent ion track hybrid detector (Cell-Fit-HD) enables the investigation of radiation-related cellular events along single ion tracks on the subcellular scale in clinical ion beams. The Cell-Fit-HD comprises a fluorescent nuclear track detector (FNTD, the physical compartment), a device for individual particle detection and a substrate for viable cell-coating, i.e. the biological compartment. To date both compartments have been imaged sequentially in situ by confocal laser scanning microscopy (CLSM). This is yet in conflict with a functional read-out of the Cell-Fit-HD utilizing a fast live-cell imaging of the biological compartment with low phototoxicity on greater time scales. The read-out of the biological from the physical compartment was uncoupled. A read-out procedure was developed to image the cell layer by conventional widefield microscopy whereas the FNTD was imaged by CLSM. Point mapping registration of the confocal and widefield imaging data was performed. Non-fluorescent crystal defects (spinels) visible in both read-outs were used as control point pairs. The accuracy achieved was on the sub-µm scale. The read-out procedure by widefield microscopy does not impair the unique ability of spatial correlation by the Cell-Fit-HD. The uncoupling will enlarge the application potential of the hybrid technology significantly. The registration allows for an ultimate correlation of microscopic physical beam parameters and cell kinetics on greater time scales. The method reported herein will be instrumental for the introduction of a novel generation of compact detectors facilitating biodosimetric research towards high-throughput analysis.

  20. Registration procedure for spatial correlation of physical energy deposition of particle irradiation and cellular response utilizing cell-fluorescent ion track hybrid detectors

    NASA Astrophysics Data System (ADS)

    Niklas, M.; Zimmermann, F.; Schlegel, J.; Schwager, C.; Debus, J.; Jäkel, O.; Abdollahi, A.; Greilich, S.

    2016-09-01

    The hybrid technology cell-fluorescent ion track hybrid detector (Cell-Fit-HD) enables the investigation of radiation-related cellular events along single ion tracks on the subcellular scale in clinical ion beams. The Cell-Fit-HD comprises a fluorescent nuclear track detector (FNTD, the physical compartment), a device for individual particle detection and a substrate for viable cell-coating, i.e. the biological compartment. To date both compartments have been imaged sequentially in situ by confocal laser scanning microscopy (CLSM). This is yet in conflict with a functional read-out of the Cell-Fit-HD utilizing a fast live-cell imaging of the biological compartment with low phototoxicity on greater time scales. The read-out of the biological from the physical compartment was uncoupled. A read-out procedure was developed to image the cell layer by conventional widefield microscopy whereas the FNTD was imaged by CLSM. Point mapping registration of the confocal and widefield imaging data was performed. Non-fluorescent crystal defects (spinels) visible in both read-outs were used as control point pairs. The accuracy achieved was on the sub-µm scale. The read-out procedure by widefield microscopy does not impair the unique ability of spatial correlation by the Cell-Fit-HD. The uncoupling will enlarge the application potential of the hybrid technology significantly. The registration allows for an ultimate correlation of microscopic physical beam parameters and cell kinetics on greater time scales. The method reported herein will be instrumental for the introduction of a novel generation of compact detectors facilitating biodosimetric research towards high-throughput analysis.

  1. Registration procedure for spatial correlation of physical energy deposition of particle irradiation and cellular response utilizing cell-fluorescent ion track hybrid detectors.

    PubMed

    Niklas, M; Zimmermann, F; Schlegel, J; Schwager, C; Debus, J; Jäkel, O; Abdollahi, A; Greilich, S

    2016-09-01

    The hybrid technology cell-fluorescent ion track hybrid detector (Cell-Fit-HD) enables the investigation of radiation-related cellular events along single ion tracks on the subcellular scale in clinical ion beams. The Cell-Fit-HD comprises a fluorescent nuclear track detector (FNTD, the physical compartment), a device for individual particle detection and a substrate for viable cell-coating, i.e. the biological compartment. To date both compartments have been imaged sequentially in situ by confocal laser scanning microscopy (CLSM). This is yet in conflict with a functional read-out of the Cell-Fit-HD utilizing a fast live-cell imaging of the biological compartment with low phototoxicity on greater time scales. The read-out of the biological from the physical compartment was uncoupled. A read-out procedure was developed to image the cell layer by conventional widefield microscopy whereas the FNTD was imaged by CLSM. Point mapping registration of the confocal and widefield imaging data was performed. Non-fluorescent crystal defects (spinels) visible in both read-outs were used as control point pairs. The accuracy achieved was on the sub-µm scale. The read-out procedure by widefield microscopy does not impair the unique ability of spatial correlation by the Cell-Fit-HD. The uncoupling will enlarge the application potential of the hybrid technology significantly. The registration allows for an ultimate correlation of microscopic physical beam parameters and cell kinetics on greater time scales. The method reported herein will be instrumental for the introduction of a novel generation of compact detectors facilitating biodosimetric research towards high-throughput analysis. PMID:27499388

  2. Improved optical transmission and current matching of a triple-junction solar cell utilizing sub-wavelength structures.

    PubMed

    Chiu, M-Y; Chang, C-H; Tsai, M-A; Chang, F-Y; Yu, Peichen

    2010-09-13

    Sub-wavelength antireflective structures are fabricated on a silicon nitride passivation layer of a Ga₀.₅In₀.₅P/GaAs/Ge triple-junction solar cell using polystyrene nanosphere lithography followed by anisotropic etching. The fabricated structures enhance optical transmission in the ultraviolet wavelength range, compared to a conventional single-layer antireflective coating (ARC). The transmission improvement contributes to an enhanced photocurrent, which is also verified by the external quantum efficiency characterization of the fabricated solar cells. Under one-sun illumination, the short-circuit current of a cell with sub-wavelength structures is enhanced by 46.1% and 3.4% due to much improved optical transmission and current matching, compared to cells without an ARC and with a conventional SiN(x) ARC, respectively. Further optimizations of the sub-wavelength structures including the periodicity and etching depth are conducted by performing comprehensive calculations based on a rigorous couple-wave analysis method.

  3. Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein.

    PubMed

    Krom, Y D; Gras, J C E; Frants, R R; Havekes, L M; van Berkel, T J; Biessen, E A L; van Dijk, K Willems

    2005-12-16

    Vascular smooth muscle (VSMC) and endothelial cells (EC) are particularly resistant to infection by type 5 adenovirus (Ad) vectors. To overcome this limitation and target Ad vectors to ubiquitously expressed alpha(V)beta(3/5) integrins, we have generated a linker protein consisting of the extracellular domain of the coxsackie adenovirus receptor (CAR) connected via avidin to a biotinylated cyclic (c) RGD peptide. After optimization of CAR to cRGD and to Ad coupling, infection of mouse heart endothelial cells (H5V) could be augmented significantly, as demonstrated by 600-fold increased transgene expression levels. In EOMAs, a hemangioendothelioma-derived cell line, the fraction of infected cells was enhanced 4- to 6-fold. Furthermore, the fraction of infected primary mouse VSMC was increased from virtually 0% to 25%. Finally, in human umbilical vein endothelial cells, the number of GFP positive cells was enhanced from 2% to 75%. In conclusion, CAR-cRGD is a versatile and highly efficient construct to target Ad vectors to both transformed and primary VSMC and EC.

  4. The Neisseria meningitidis ZnuD Zinc Receptor Contributes to Interactions with Epithelial Cells and Supports Heme Utilization when Expressed in Escherichia coli

    PubMed Central

    Kumar, Pradeep; Sannigrahi, Soma

    2012-01-01

    Neisseria meningitidis employs redundant heme acquisition mechanisms, including TonB receptor-dependent and receptor-independent uptakes. The TonB-dependent zinc receptor ZnuD shares significant sequence similarity to HumA, a heme receptor of Moraxella catarrhalis, and contains conserved motifs found in many heme utilization proteins. We present data showing that, when expressed in Escherichia coli, ZnuD allowed heme capture on the cell surface and supported the heme-dependent growth of an E. coli hemA strain. Heme agarose captured ZnuD in enriched outer membrane fractions, and this binding was inhibited by excess free heme, supporting ZnuD's specific interaction with heme. However, no heme utilization defect was detected in the meningococcal znuD mutant, likely due to unknown redundant TonB-independent heme uptake mechanisms. Meningococcal replication within epithelial cells requires a functional TonB, and we found that both the znuD and tonB mutants were defective not only in survival within epithelial cells but also in adherence to and invasion of epithelial cells. Ectopic complementation rescued these phenotypes. Interestingly, while znuD expression was repressed by Zur with zinc as a cofactor, it also was induced by iron in a Zur-independent manner. A specific interaction of meningococcal Fur protein with the znuD promoter was demonstrated by electrophoretic mobility shift assay (EMSA). Thus, the meningococcal ZnuD receptor likely participates in both zinc and heme acquisition, is regulated by both Zur and Fur, and is important for meningococcal interaction with epithelial cells. PMID:22083713

  5. Investigation of the effects of external current systems on the MAGSAT data utilizing grid cell modeling techniques

    NASA Technical Reports Server (NTRS)

    Klumpar, D. M. (Principal Investigator)

    1982-01-01

    Progress made in reducing MAGSAT data and displaying magnetic field perturbations caused primarily by external currents is reported. A periodic and repeatable perturbation pattern is described that arises from external current effects but appears as unique signatures associated with upper middle latitudes on the Earth's surface. Initial testing of the modeling procedure that was developed to compute the magnetic fields at satellite orbit due to current distributions in the ionosphere and magnetosphere is also discussed. The modeling technique utilizes a linear current element representation of the large scale space current system.

  6. Utilizing induced pluripotent stem cells (iPSCs) to understand the actions of estrogens in human neurons.

    PubMed

    Shum, Carole; Macedo, Sara C; Warre-Cornish, Katherine; Cocks, Graham; Price, Jack; Srivastava, Deepak P

    2015-08-01

    This article is part of a Special Issue "Estradiol and Cognition". Over recent years tremendous progress has been made towards understanding the molecular and cellular mechanism by which estrogens exert enhancing effects on cognition, and how they act as a neuroprotective or neurotrophic agent in disease. Currently, much of this work has been carried out in animal models with only a limited number of studies using native human tissue or cells. Recent advances in stem cell technology now make it possible to reprogram somatic cells from humans into induced pluripotent stem cells (iPSCs), which can subsequently be differentiated into neurons of specific lineages. Importantly, the reprogramming of cells allows for the generation of iPSCs that retain the genetic "makeup" of the donor. Therefore, it is possible to generate iPSC-derived neurons from patients diagnosed with specific diseases, that harbor the complex genetic background associated with the disorder. Here, we review the iPSC technology and how it's currently being used to model neural development and neurological diseases. Furthermore, we explore whether this cellular system could be used to understand the role of estrogens in human neurons, and present preliminary data in support of this. We further suggest that the use of iPSC technology offers a novel system to not only further understand estrogens' effects in human cells, but also to investigate the mechanism by which estrogens are beneficial in disease. Developing a greater understanding of these mechanisms in native human cells will also aid in the development of safer and more effective estrogen-based therapeutics. PMID:26143621

  7. Utilization of Cheese Whey Using Synergistic Immobilization of β-Galactosidase and Saccharomyces cerevisiae Cells in Dual Matrices.

    PubMed

    Kokkiligadda, Anusha; Beniwal, Arun; Saini, Priyanka; Vij, Shilpa

    2016-08-01

    Whey is a byproduct of the dairy industry, which has prospects of using as a source for production of various valuable compounds. The lactose present in whey is considered as an environmental pollutant and its utilization for enzyme and fuel production, may be effective for whey bioremediation. The dairy yeast Kluyveromyces marxianus have the ability to utilize lactose sharply as the major carbon source for the production of the enzyme. Five strains were tested for the production of the β-galactosidase using whey. The maximum β-galactosidase activity of 1.74 IU/mg dry weight was achieved in whey using K. marxianus MTCC 1389. The biocatalyst was further immobilized on chitosan macroparticles and exhibited excellent functional activity at 35 °C. Almost 89 % lactose hydrolysis was attained for concentrated whey (100 g/L) and retained 89 % catalytic activity after 15 cycles of reuse. Finally, β-galactosidase was immobilized on chitosan and Saccharomyces cerevisiae on calcium alginate, and both were used together for the production of ethanol from concentrated whey. Maximal ethanol titer of 28.9 g/L was achieved during fermentation at 35 °C. The conclusions generated by employing two different matrices will be beneficial for the future modeling using engineered S. cerevisiae in scale-up studies. PMID:27059625

  8. Utilization of Cheese Whey Using Synergistic Immobilization of β-Galactosidase and Saccharomyces cerevisiae Cells in Dual Matrices.

    PubMed

    Kokkiligadda, Anusha; Beniwal, Arun; Saini, Priyanka; Vij, Shilpa

    2016-08-01

    Whey is a byproduct of the dairy industry, which has prospects of using as a source for production of various valuable compounds. The lactose present in whey is considered as an environmental pollutant and its utilization for enzyme and fuel production, may be effective for whey bioremediation. The dairy yeast Kluyveromyces marxianus have the ability to utilize lactose sharply as the major carbon source for the production of the enzyme. Five strains were tested for the production of the β-galactosidase using whey. The maximum β-galactosidase activity of 1.74 IU/mg dry weight was achieved in whey using K. marxianus MTCC 1389. The biocatalyst was further immobilized on chitosan macroparticles and exhibited excellent functional activity at 35 °C. Almost 89 % lactose hydrolysis was attained for concentrated whey (100 g/L) and retained 89 % catalytic activity after 15 cycles of reuse. Finally, β-galactosidase was immobilized on chitosan and Saccharomyces cerevisiae on calcium alginate, and both were used together for the production of ethanol from concentrated whey. Maximal ethanol titer of 28.9 g/L was achieved during fermentation at 35 °C. The conclusions generated by employing two different matrices will be beneficial for the future modeling using engineered S. cerevisiae in scale-up studies.

  9. A morphological study of the pacemaker cells of the aganglionic intestine in Hirschsprung's disease utilizing ls/ls model mice.

    PubMed

    Taniguchi, Kan; Matsuura, Kimio; Matsuoka, Takanori; Nakatani, Hajime; Nakano, Takumi; Furuya, Yasuo; Sugimoto, Takeki; Kobayashi, Michiya; Araki, Keijiro

    2005-06-01

    Hirschsprung's disease is a congenital aganglionic neural disorder of the segmental distal intestine characterized by unsettled pathogenesis. The relationship between Hirschsprung's disease and pacemaker cells (PMC), which almost corresponds to that of the interstitial cells of Cajal (ICC), was morphologically observed at the level of the intermuscular layer corresponding to Auerbach's plexus using ls/ls mice. These mice are an ideal model because of their large intestinal aganglionosis and gene abnormalities, which are similar to the human form of the disease. Immunostaining using anti-c-kit receptor antibody (ACK2), a marker of PMC, applied to whole-mount muscle-layer specimens, revealed the presence of c-kit immunopositive multipolar cells with many cytoplasmic processes in normal mice. For ls/ls mice, however, there were significantly fewer processes. The average number of processes per positive cell of 2.5 for the aganglionic large intestine was fewer than 3.5 for the large and small intestine of normal mice, indicating the inability to form connections between nerves and PMC in the aganglionic intestine. For normal mice with an Auerbach's plexus, the process attachment of ICC to the Auerbach's plexus was observed by scanning electron microscopy. However, for ls/ls mice no attachment to the intermuscular nerve without Auerbach's plexus was found, although transmission electron microscopy showed no difference in the cell structure and organelles of the c-kit immunopositive cells between the normal and ls/ls mice. These findings suggest that in the aganglionic intestine of Hirschsprung's disease, aplasia of enteric ganglia induces secondary disturbances during the normal development of intestinal PMC.

  10. Utilization of arsenic trioxide as a treatment of cisplatin-resistant non-small cell lung cancer PC-9/CDDP and PC-14/CDDP cells

    PubMed Central

    SUZUKI, TOSHIHIRO; ISHIBASHI, KENICHI; YUMOTO, ATSUSHI; NISHIO, KAZUTO; OGASAWARA, YUKI

    2015-01-01

    Cisplatin is a commonly used drug in combination chemotherapy. However, various malignant tumors frequently acquire resistance to cisplatin. Arsenic trioxide (ATO) has been approved as a chemotherapeutic drug for the treatment of acute promyelocytic leukemia, and the combination of ATO and cisplatin has been revealed to demonstrate synergistic effects in ovarian and small cell lung cancer cells. Thus, it was hypothesized that ATO may also be active against cisplatin-resistant non-small cell lung cancer (NSCLC) PC-9/CDDP and PC-14/CDDP cells. The present study also evaluated the effects of ATO on the cisplatin-sensitive NSCLC PC-9 and PC-14 cell lines. Notably, ATO demonstrated a markedly decreased IC50 in the cisplatin-resistant PC-9/CDDP and PC-14/CDDP cells compared with the IC50 in the cisplatin-sensitive parental PC-9 and PC-14 cells. Additionally, it was found that arsenite accumulation in the PC-9 cell line was affected through the downregulation of GS-X pump systems. Although it is likely that cisplatin resistance in PC-9 cells does not depend on the GS-X pump systems, ATO was effective against cisplatin-resistant NSCLC PC-9/CDDP and PC-14/CDDP cells in combination chemotherapy. PMID:26622574

  11. Catalytic modification of Ni-Sm-doped ceria anodes with copper for direct utilization of dry methane in low-temperature solid oxide fuel cells

    NASA Astrophysics Data System (ADS)

    Wang, Zhicheng; Weng, Wenjian; Cheng, Kui; Du, Piyi; Shen, Ge; Han, Gaorong

    2008-05-01

    A Cu/Ni/Sm-doped ceria (SDC) anode has been designed for direct utilization of dry methane in low-temperature anode-supported solid oxide fuel cells. The anode is prepared by the impregnation method, whereby a small amount of Cu is incorporated into the previously prepared Ni/SDC porous matrix. After reduction, Cu nanoparticles adhere to and are uniformly distributed on the surface of the Ni/SDC matrix. For the resulting Cu/Ni/SDC anode-supported cell, maximum power density of 317 mW cm-2 is achieved at 600 °C. The power density shows only ∼2% loss after 12-h operation. The results demonstrate that the Cu/Ni/SDC anode effectively suppresses carbon deposition by decreasing the Ni surface area available and the level of carbon monoxide disproportionation. This combination of effects results in very low-power density loss over the operating time.

  12. Efficient biological conversion of carbon monoxide (CO) to carbon dioxide (CO2) and for utilization in bioplastic production by Ralstonia eutropha through the display of an enzyme complex on the cell surface.

    PubMed

    Hyeon, Jeong Eun; Kim, Seung Wook; Park, Chulhwan; Han, Sung Ok

    2015-06-25

    An enzyme complex for biological conversion of CO to CO2 was anchored on the cell surface of the CO2-utilizing Ralstonia eutropha and successfully resulted in a 3.3-fold increase in conversion efficiency. These results suggest that this complexed system may be a promising strategy for CO2 utilization as a biological tool for the production of bioplastics. PMID:26017299

  13. Efficient biological conversion of carbon monoxide (CO) to carbon dioxide (CO2) and for utilization in bioplastic production by Ralstonia eutropha through the display of an enzyme complex on the cell surface.

    PubMed

    Hyeon, Jeong Eun; Kim, Seung Wook; Park, Chulhwan; Han, Sung Ok

    2015-06-25

    An enzyme complex for biological conversion of CO to CO2 was anchored on the cell surface of the CO2-utilizing Ralstonia eutropha and successfully resulted in a 3.3-fold increase in conversion efficiency. These results suggest that this complexed system may be a promising strategy for CO2 utilization as a biological tool for the production of bioplastics.

  14. Utilizing Viral Nanoparticle/Dendron Hybrid Conjugates in Photodynamic Therapy for Dual Delivery to Macrophages and Cancer Cells.

    PubMed

    Wen, Amy M; Lee, Karin L; Cao, Pengfei; Pangilinan, Katrina; Carpenter, Bradley L; Lam, Patricia; Veliz, Frank A; Ghiladi, Reza A; Advincula, Rigoberto C; Steinmetz, Nicole F

    2016-05-18

    Photodynamic therapy (PDT) is a promising avenue for greater treatment efficacy of highly resistant and aggressive melanoma. Through photosensitizer attachment to nanoparticles, specificity of delivery can be conferred to further reduce potential side effects. While the main focus of PDT is the destruction of cancer cells, additional targeting of tumor-associated macrophages also present in the tumor microenvironment could further enhance treatment by eliminating their role in processes such as invasion, metastasis, and immunosuppression. In this study, we investigated PDT of macrophages and tumor cells through delivery using the natural noninfectious nanoparticle cowpea mosaic virus (CPMV), which has been shown to have specificity for the immunosuppressive subpopulation of macrophages and also targets cancer cells. We further explored conjugation of CPMV/dendron hybrids in order to improve the drug loading capacity of the nanocarrier. Overall, we demonstrated effective elimination of both macrophage and tumor cells at low micromolar concentrations of the photosensitizer when delivered with the CPMV bioconjugate, thereby potentially improving melanoma treatment. PMID:27077475

  15. Using biomimetic cell wall models to identify new plant lignin bioengineering targets for improving forage and biomass utilization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bioengineering of lignin to contain atypical components derived from other metabolic pathways is increasingly being pursued to custom design lignified cell walls that are inherently more digestible by livestock or more easily pretreated and saccharified for biofuel production. Because plants produce...

  16. Identifying new lignin bioengineering targets for improving biomass and forage utilization: a review of biomimetic studies with maize cell walls

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bioengineering of lignin to contain atypical components derived from other metabolic pathways is increasingly being pursued to custom design lignified cell walls that are more readily pretreated and saccharified for biofuel production or easily digested by livestock. Because plants produce such a di...

  17. Limitations and relative utility of screening assays to assess engineered nanoparticle toxicity in a human cell line

    SciTech Connect

    Monteiro-Riviere, N.A.; Inman, A.O.; Zhang, L.W.

    2009-01-15

    Single-walled carbon nanotubes (SWCNT), fullerenes (C{sub 60}), carbon black (CB), nC{sub 60}, and quantum dots (QD) have been studied in vitro to determine their toxicity in a number of cell types. Here, we report that classical dye-based assays such as MTT and neutral red (NR) that determine cell viability produce invalid results with some NM (nanomaterials) due to NM/dye interactions and/or NM adsorption of the dye/dye products. In this study, human epidermal keratinocytes (HEK) were exposed in vitro to CB, SWCNT, C{sub 60}, nC{sub 60}, and QD to assess viability with calcein AM (CAM), Live/Dead (LD), NR, MTT, Celltiter 96 AQueous One (96 AQ), alamar Blue (aB), Celltiter-Blue (CTB), CytoTox One{sup TM} (CTO), and flow cytometry. In addition, trypan blue (TB) was quantitated by light microscopy. Assay linearity (R{sup 2} value) was determined with HEK plated at concentrations from 0 to 25,000 cells per well in 96-well plates. HEK were treated with serial dilutions of each NM for 24 h and assessed with each of the viability assays. TB, CAM and LD assays, which depend on direct staining of living and/or dead cells, were difficult to interpret due to physical interference of the NM with cells. Results of the dye-based assays varied a great deal, depending on the interactions of the dye/dye product with the carbon nanomaterials (CNM). Results show the optimal high throughput assay for use with carbon and noncarbon NM was 96 AQ. This study shows that, unlike small molecules, CNM interact with assay markers to cause variable results with classical toxicology assays and may not be suitable for assessing nanoparticle cytotoxicity. Therefore, more than one assay may be required when determining nanoparticle toxicity for risk assessment.

  18. Enhanced plasmid DNA utilization in transiently transfected CHO-DG44 cells in the presence of polar solvents.

    PubMed

    Rajendra, Yashas; Balasubramanian, Sowmya; Kiseljak, Divor; Baldi, Lucia; Wurm, Florian M; Hacker, David L

    2015-01-01

    Although the protein yields from transient gene expression (TGE) with Chinese hamster ovary (CHO) cells have recently improved, the amount of plasmid DNA (pDNA) needed for transfection remains relatively high. We describe a strategy to reduce the pDNA amount by transfecting CHO-DG44 cells with 0.06 μg pDNA/10(6) cells (10% of the optimal amount) in the presence of nonspecific (filler) DNA and various polar solvents including dimethylsufoxide, dimethyl formamide, acetonitrile, dimethyl acetamide (DMA), and hexamethyl phosphoramide (HMP). All of the polar solvents with the exception of HMP increased the production of a recombinant antibody in comparison to the untreated control transfection. In the presence of 0.25% DMA, the antibody yield in a 7-day batch culture was 500 mg/L. This was fourfold higher than the yield from the untreated control transfection. Mechanistic studies revealed that the polar solvents did not affect polyethylenimine-mediated pDNA delivery into cells or nuclei. The steady-state transgene mRNA level was elevated in the presence of each of the polar solvents tested, while the transgene mRNA half-life remained the same. These results indicated that the polar solvents enhanced transgene transcription. When screening a panel of recombinant antibodies and Fc-fusion proteins for production in the presence of the polar solvents, the highest increase in yield was observed following DMA addition for 11 of the 12 proteins. These results are expected to enhance the applicability of high-yielding TGE processes with CHO-DG44 cells by decreasing the amount of pDNA required for transfection.

  19. Utility of large-scale transiently transfected cells for cell-based high-throughput screens to identify transient receptor potential channel A1 (TRPA1) antagonists.

    PubMed

    Chen, Jun; Lake, Marc R; Sabet, Reza S; Niforatos, Wende; Pratt, Steve D; Cassar, Steven C; Xu, Jing; Gopalakrishnan, Sujatha; Pereda-Lopez, Ana; Gopalakrishnan, Murali; Holzman, Thomas F; Moreland, Robert B; Walter, Karl A; Faltynek, Connie R; Warrior, Usha; Scott, Victoria E

    2007-02-01

    Despite increasing use of cell-based assays in high-throughput screening (HTS) and lead optimization, one challenge is the adequate supply of high-quality cells expressing the target of interest. To this end, cell lines stably expressing targets are often established, maintained, and scaled up by cell culture. These steps require large investments of time and resources. Moreover, significant variability invariably occurs in cell yield, viability, expression levels, and target activities. In particular, stable expression of targets such as transient receptor potential A1 (TRPA1) causes toxicity, cell line degeneration, and loss of functional activity. Therefore, in an effort to identify TRPA1 antagonists, the authors used large-scale transiently transfected (LSTT) cells, enabling rapid establishment of assays suitable for HTS. LSTT cells, which could- be stored frozen for a long period of time (e.g., at least 42 weeks), retained TRPA1 protein expression and could be easily revived to produce robust and consistent signals in calcium influx and electrophysiological assays. Using cells from a single transfection, a chemical library of 700,000 compounds was screened, and TRPA1 antagonists were identified. The use of LSTT circumvented issues associated with stable TRPA1 expression, increased flexibility and consistency, and greatly reduced labor and cost. This approach will also be applicable to other pharmaceutical targets.

  20. Energy filtering transmission electron microscopy immunocytochemistry and antigen retrieval of surface layer proteins from Tannerella forsythensis using microwave or autoclave heating with citraconic anhydride.

    PubMed

    Moriguchi, K; Mitamura, Y; Iwami, J; Hasegawa, Y; Higuchi, N; Murakami, Y; Maeda, H; Yoshimura, F; Nakamura, H; Ohno, N

    2012-11-01

    Tannerella forsythensis (Bacteroides forsythus), an anaerobic Gram-negative species of bacteria that plays a role in the progression of periodontal disease, has a unique bacterial protein profile. It is characterized by two unique protein bands with molecular weights of more than 200 kDa. It also is known to have a typical surface layer (S-layer) consisting of regularly arrayed subunits outside the outer membrane. We examined the relationship between high molecular weight proteins and the S-layer using electron microscopic immunolabeling with chemical fixation and an antigen retrieval procedure consisting of heating in a microwave oven or autoclave with citraconic anhydride. Immunogold particles were localized clearly at the outermost cell surface. We also used energy-filtering transmission electron microscopy (EFTEM) to visualize 3, 3'-diaminobenzidine tetrahydrochloride (DAB) reaction products after microwave antigen retrieval with 1% citraconic anhydride. The three-window method for electron spectroscopic images (ESI) of nitrogen by the EFTEM reflected the presence of moieties demonstrated by the DAB reaction with horseradish peroxidase (HRP)-conjugated secondary antibodies instead of immunogold particles. The mapping patterns of net nitrogen were restricted to the outermost cell surface.

  1. Improved performance of P3HT:PCBM solar cells by both anode modification and short-wavelength energy utilization using Tb(aca)3phen

    NASA Astrophysics Data System (ADS)

    Zhuo, Zu-Liang; Wang, Yong-Sheng; He, Da-Wei; Fu, Ming

    2014-09-01

    The performance of P3HT:PCBM solar cells was improved by anode modification using spin-coated Tb(aca)3phen ultrathin films. The modification of the Tb(aca)3phen ultrathin film between the indium tin oxide (ITO) anode and the PE-DOT:PSS layer resulted in a maximum power conversion efficiency (PCE) of 2.99% compared to 2.66% for the reference device, which was due to the increase in the short-circuit current density (Jsc). The PCE improvement could be attributed to the short-wavelength energy utilization and the optimized morphology of the active layers. Tb(aca)3phen with its strong down-conversion luminescence properties is suitable for the P3HT:PCBM blend active layer, and the absorption region of the ternary blend films is extended into the near ultraviolet region. Furthermore, the crystallization and the surface morphology of P3HT:PCBM films were improved with the Tb(aca)3phen ultrathin film. The ultraviolent—visible absorption spectra, atomic force microscope (AFM), and X-ray diffraction (XRD) of the films were investigated. Both anode modification and short-wavelength energy utilization using Tb(aca)3phen in P3HT:PCBM solar cells led to about a 12% PCE increase.

  2. Coordinated regulation of nitrogen supply mode and initial cell density for energy storage compounds production with economized nitrogen utilization in a marine microalga Isochrysis zhangjiangensis.

    PubMed

    Chi, Lei; Yao, Changhong; Cao, Xupeng; Xue, Song

    2016-01-01

    Lipids and carbohydrates are main energy storage compounds (ESC) of microalgae under stressed conditions and they are potential feedstock for biofuel production. Yet, the sustainable and commercially successful production of ESC in microalgae needs to consider nitrogen utilization efficiency. Here the impact of different initial cell densities (ICDs) on ESC accumulation in Isochrysis zhangjiangensis under two nitrogen supply modes (an initially equal concentration of nitrogen per-cell in the medium (N1) and an equal total concentration of nitrogen in the culture system (N2)) were investigated. The results demonstrated that the highest ESC yield (1.36gL(-1)) at N1, which included a maximal nitrogen supply in the cultivation system, and the highest ESC content (66.5%) and ESC productivity per mass of nitrogen (3.28gg(-1) (N) day(-1)) at N2, were all obtained under a high ICD of 8.0×10(6)cellsmL(-1). Therefore I. zhangjiangensis qualifies for ESC-enriched biomass production with economized nitrogen utilization.

  3. Utilization of Pt/Ru catalysts in MEA for fuel cell application by breathing process of proton exchange membrane

    NASA Astrophysics Data System (ADS)

    Yu, Kyung-Chul; Kim, Woo-Jae; Chung, Chan-Hwa

    Small direct-methanol fuel cells (DMFCs) have recently been highlighted as possible power sources for applications ranging from cellular phones and wireless digital devices to autonomous sensors and micro-electro mechanical devices. One of the key issues in commercializing miniaturized DMFCs for portable applications is to improve the electrochemical performance of the cells with a small quantity of catalysts. Up to now, the spraying or brush method has been used to fabricate a catalyst layer, which uses a slurry of nano-sized Pt or Pt/Ru catalysts. However, these methods produce a poor electrochemical interface that reduces the catalytic activity and the reproducibility of their performance tests. In this study, a unique process known as a "breathing process" was used to fabricate a catalytic electrode layer in a membrane-electrode-assembly (MEA) of DMFCs. The Pt/Ru nano-particles were loaded directly onto a proton exchange membrane using this breathing process. This process consisted of the following three steps: (1) the electrolyte membrane was fully swollen in water; (2) the swollen membrane was placed into an aprotic solvent, which induced the shrinkage of the membrane by driving the water out ("breathing out"); (3) the shrunken membrane was placed in an aqueous solution containing a suspension of Pt/Ru nano-particles. This induces the swelling of the membrane, and the suspended Pt/Ru nano-particles penetrate the membrane during this process ("breathing in"). It is possible to control the amount of catalysts loaded in the MEA by controlling the number of the cycles of such breathing processes. Compared with the fuel cell adopting the MEA fabricated by a conventional spraying method with the same amount of catalysts, the performance of this novel fuel cell was enhanced by approximately 4.5 mW cm -2 in case of the passive-type fuel cell and by 9.0 mW cm -2 in case of the active-type. This paper details the optimized process conditions along with other advanced

  4. Evaluation of the Paratrend Multi-Analyte Sensor for Potential Utilization in Long-Duration Automated Cell Culture Monitoring

    NASA Technical Reports Server (NTRS)

    Hwang, Emma Y.; Pappas, Dimitri; Jeevarajan, Antony S.; Anderson, Melody M.

    2004-01-01

    BACKGROUND: Compact and automated sensors are desired for assessing the health of cell cultures in biotechnology experiments. While several single-analyte sensors exist to measure culture health, a multi-analyte sensor would simplify the cell culture system. One such multi-analyte sensor, the Paratrend 7 manufactured by Diametrics Medical, consists of three optical fibers for measuring pH, dissolved carbon dioxide (pCO(2)), dissolved oxygen (pO(2)), and a thermocouple to measure temperature. The sensor bundle was designed for intra-vascular measurements in clinical settings, and can be used in bioreactors operated both on the ground and in NASA's Space Shuttle and International Space Station (ISS) experiments. METHODS: A Paratrend 7 sensor was placed at the outlet of a bioreactor inoculated with BHK-21 (baby hamster kidney) cells. The pH, pCO(2), pO(2), and temperature data were transferred continuously to an external computer. Cell culture medium, manually extracted from the bioreactor through a sampling port, was also assayed using a bench top blood gas analyzer (BGA). RESULTS: Two Paratrend 7 sensors were used over a single cell culture experiment (64 days). When compared to the manually obtained BGA samples, the sensor had good agreement for pH, pCO(2), and pO(2) with bias (and precision) 0.005(0.024), 8.0 mmHg (4.4 mmHg), and 11 mmHg (17 mmHg), respectively for the first two sensors. A third Paratrend sensor (operated for 141 days) had similar agreement (0.02+/-0.15 for pH, -4+/-8 mm Hg for pCO(2), and 24+/-18 mmHg for pO(2)). CONCLUSION: The resulting biases and precisions are com- parable to Paratrend sensor clinical results. Although the pO(2) differences may be acceptable for clinically relevant measurement ranges, the O(2) sensor in this bundle may not be reliable enough for the ranges of pO(2) in these cell culture studies without periodic calibration.

  5. Cost utility analysis of reduced intensity hematopoietic stem cell transplantation in adolescence and young adult with severe thalassemia compared to hypertransfusion and iron chelation program

    PubMed Central

    2013-01-01

    Background Hematopoieticic stem cell transplantation is the only therapeutic option that can cure thalassemia disease. Reduced intensity hematopoietic stem cell transplantation (RI-HSCT) has demonstrated a high cure rate with minimal complications compared to other options. Because RI-HSCT is very costly, economic justification for its value is needed. This study aimed to estimate the cost-utility of RI-HSCT compared with blood transfusions combined with iron chelating therapy (BT-ICT) for adolescent and young adult with severe thalassemia in Thailand. Methods A Markov model was used to estimate the relevant costs and health outcomes over the patients’ lifetimes using a societal perspective. All future costs and outcomes were discounted at a rate of 3% per annum. The efficacy of RI-HSCT was based a clinical trial including a total of 18 thalassemia patients. Utility values were derived directly from all patients using EQ-5D and SF-6D. Primary outcomes of interest were lifetime costs, quality adjusted life-years (QALYs) gained, and the incremental cost-effectiveness ratio (ICER) in US ($) per QALY gained. One-way and probabilistic sensitivity analyses (PSA) were conducted to investigate the effect of parameter uncertainty. Results In base case analysis, the RI-HSCT group had a better clinical outcomes and higher lifetime costs. The incremental cost per QALY gained was US $ 3,236 per QALY. The acceptability curve showed that the probability of RI-HSCT being cost-effective was 71% at the willingness to pay of 1 time of Thai Gross domestic product per capita (GDP per capita), approximately US $ 4,210 per QALY gained. The most sensitive parameter was utility of severe thalassemia patients without cardiac complication patients. Conclusion At a societal willingness to pay of 1 GDP per capita, RI-HSCT was a cost-effective treatment for adolescent and young adult with severe thalassemia in Thailand compared to BT-ICT. PMID:23379888

  6. Campylobacter jejuni carbon starvation protein A (CstA) is involved in peptide utilization, motility and agglutination, and has a role in stimulation of dendritic cells.

    PubMed

    Rasmussen, J J; Vegge, C S; Frøkiær, H; Howlett, R M; Krogfelt, K A; Kelly, D J; Ingmer, H

    2013-08-01

    Campylobacter jejuni is the most frequent cause of severe gastroenteritis in the developed world. The major symptom of campylobacteriosis is inflammatory diarrhoea. The molecular mechanisms of this infection are poorly understood compared to those of less frequent disease-causing pathogens. In a previous study, we identified C. jejuni proteins that antibodies in human campylobacteriosis patients reacted with. One of the immunogenic proteins identified (Cj0917) displays homology to carbon starvation protein A (CstA) from Escherichia coli, where this protein is involved in the starvation response and peptide uptake. In contrast to many bacteria, C. jejuni relies on amino acids and organic acids for energy, but in vivo it is highly likely that peptides are also utilized, although their mechanisms of uptake are unknown. In this study, Biolog phenotype microarrays have been used to show that a ΔcstA mutant has a reduced ability to utilize a number of di- and tri-peptides as nitrogen sources. This phenotype was restored through genetic complementation, suggesting CstA is a peptide uptake system in C. jejuni. Furthermore, the ΔcstA mutant also displayed reduced motility and reduced agglutination compared to WT bacteria; these phenotypes were also restored through complementation. Murine dendritic cells exposed to UV-killed bacteria showed a reduced IL-12 production, but the same IL-10 response when encountering C. jejuni ΔcstA compared to the WT strain. The greater Th1 stimulation elicited by the WT as compared to ΔcstA mutant cells indicates an altered antigenic presentation on the surface, and thus an altered recognition of the mutant. Thus, we conclude that C. jejuni CstA is important not only for peptide utilization, but also it may influence host-pathogen interactions. PMID:23682166

  7. Lighting Utilization.

    ERIC Educational Resources Information Center

    Crank, Ron

    This instructional unit is one of 10 developed by students on various energy-related areas that deals specifically with lighting utilization. Its objective is for the student to be able to outline the development of lighting use and conservation and identify major types and operating characteristics of lamps used in electric lighting. Some topics…

  8. Selection of Variants Utilizing Heparin Sulphate For Cell Entry When South African Territories Foot-and-Mouth Disease Virus is Adapted for Growth on Cell Culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Foot-and-mouth disease virus (FMDV) attains entry to epithelial cells by affinity for at least four members of the integrin family of receptors. Adaptation of field isolates to grow in cultured cells is an essential step towards development of vaccines against new outbreak strains. This is made poss...

  9. Method of fabricating conducting oxide-silicon solar cells utilizing electron beam sublimation and deposition of the oxide

    DOEpatents

    Feng, Tom; Ghosh, Amal K.

    1979-01-01

    In preparing tin oxide and indium tin oxide-silicon heterojunction solar cells by electron beam sublimation of the oxide and subsequent deposition thereof on the silicon, the engineering efficiency of the resultant cell is enhanced by depositing the oxide at a predetermined favorable angle of incidence. Typically the angle of incidence is between 40.degree. and 70.degree. and preferably between 55.degree. and 65.degree. when the oxide is tin oxide and between 40.degree. and 70.degree. when the oxide deposited is indium tin oxide. gi The Government of the United States of America has rights in this invention pursuant to Department of Energy Contract No. EY-76-C-03-1283.

  10. Acetyl-CoA Synthetase 2 Promotes Acetate Utilization and Maintains Cancer Cell Growth under Metabolic Stress

    PubMed Central

    Schug, Zachary T.; Peck, Barrie; Jones, Dylan T.; Zhang, Qifeng; Grosskurth, Shaun; Alam, Israt S.; Goodwin, Louise M.; Smethurst, Elizabeth; Mason, Susan; Blyth, Karen; McGarry, Lynn; James, Daniel; Shanks, Emma; Kalna, Gabriela; Saunders, Rebecca E.; Jiang, Ming; Howell, Michael; Lassailly, Francois; Thin, May Zaw; Spencer-Dene, Bradley; Stamp, Gordon; van den Broek, Niels J.F.; Mackay, Gillian; Bulusu, Vinay; Kamphorst, Jurre J.; Tardito, Saverio; Strachan, David; Harris, Adrian L.; Aboagye, Eric O.; Critchlow, Susan E.; Wakelam, Michael J.O.; Schulze, Almut; Gottlieb, Eyal

    2015-01-01

    Summary A functional genomics study revealed that the activity of acetyl-CoA synthetase 2 (ACSS2) contributes to cancer cell growth under low-oxygen and lipid-depleted conditions. Comparative metabolomics and lipidomics demonstrated that acetate is used as a nutritional source by cancer cells in an ACSS2-dependent manner, and supplied a significant fraction of the carbon within the fatty acid and phospholipid pools. ACSS2 expression is upregulated under metabolically stressed conditions and ACSS2 silencing reduced the growth of tumor xenografts. ACSS2 exhibits copy-number gain in human breast tumors, and ACSS2 expression correlates with disease progression. These results signify a critical role for acetate consumption in the production of lipid biomass within the harsh tumor microenvironment. PMID:25584894

  11. Spatial and Temporal Analysis of Human Movements and Applications for Disaster Response Management Utilizing Cell Phone Usage Data

    NASA Astrophysics Data System (ADS)

    Yasumiishi, M.; Renschler, C. S.; Bittner, T. E.

    2015-07-01

    As cell phone usage becomes a norm in our daily lives, analysis and application of the data has become part of various research fields. This study focuses on the application of cell phone usage data to disaster response management. Cell phones work as a communication link between emergency responders and victims during and after a major disaster. This study recognizes that there are two kinds of disasters, one with an advance warning, and one without an advance warning. Different movement distance between a day with a blizzard (advanced warning) and a normal weather day was identified. In the scenario of a day with an extreme event without advanced warning (earthquake), factors that alter the phone users' movements were analyzed. Lastly, combining both cases, a conceptual model of human movement factors is proposed. Human movements consist of four factors that are push factors, movement-altering factors, derived attributes and constraint factors. Considering each category of factors in case of emergency, it should be necessary that we prepare different kinds of emergency response plans depending on the characteristics of a disaster.

  12. Microbial surface displayed enzymes based biofuel cell utilizing degradation products of lignocellulosic biomass for direct electrical energy.

    PubMed

    Fan, Shuqin; Hou, Chuantao; Liang, Bo; Feng, Ruirui; Liu, Aihua

    2015-09-01

    In this work, a bacterial surface displaying enzyme based two-compartment biofuel cell for the direct electrical energy conversion from degradation products of lignocellulosic biomass is reported. Considering that the main degradation products of the lignocellulose are glucose and xylose, xylose dehydrogenase (XDH) displayed bacteria (XDH-bacteria) and glucose dehydrogenase (GDH) displayed bacteria (GDH-bacteria) were used as anode catalysts in anode chamber with methylene blue as electron transfer mediator. While the cathode chamber was constructed with laccase/multi-walled-carbon nanotube/glassy-carbon-electrode. XDH-bacteria exhibited 1.75 times higher catalytic efficiency than GDH-bacteria. This assembled enzymatic fuel cell exhibited a high open-circuit potential of 0.80 V, acceptable stability and energy conversion efficiency. Moreover, the maximum power density of the cell could reach 53 μW cm(-2) when fueled with degradation products of corn stalk. Thus, this finding holds great potential to directly convert degradation products of biomass into electrical energy. PMID:26051524

  13. Microbial surface displayed enzymes based biofuel cell utilizing degradation products of lignocellulosic biomass for direct electrical energy.

    PubMed

    Fan, Shuqin; Hou, Chuantao; Liang, Bo; Feng, Ruirui; Liu, Aihua

    2015-09-01

    In this work, a bacterial surface displaying enzyme based two-compartment biofuel cell for the direct electrical energy conversion from degradation products of lignocellulosic biomass is reported. Considering that the main degradation products of the lignocellulose are glucose and xylose, xylose dehydrogenase (XDH) displayed bacteria (XDH-bacteria) and glucose dehydrogenase (GDH) displayed bacteria (GDH-bacteria) were used as anode catalysts in anode chamber with methylene blue as electron transfer mediator. While the cathode chamber was constructed with laccase/multi-walled-carbon nanotube/glassy-carbon-electrode. XDH-bacteria exhibited 1.75 times higher catalytic efficiency than GDH-bacteria. This assembled enzymatic fuel cell exhibited a high open-circuit potential of 0.80 V, acceptable stability and energy conversion efficiency. Moreover, the maximum power density of the cell could reach 53 μW cm(-2) when fueled with degradation products of corn stalk. Thus, this finding holds great potential to directly convert degradation products of biomass into electrical energy.

  14. Utilization of Whole-Cell MALDI-TOF Mass Spectrometry to Differentiate Burkholderia pseudomallei Wild-Type and Constructed Mutants.

    PubMed

    Niyompanich, Suthamat; Srisanga, Kitima; Jaresitthikunchai, Janthima; Roytrakul, Sittiruk; Tungpradabkul, Sumalee

    2015-01-01

    Whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (whole-cell MALDI-TOF MS) has been widely adopted as a useful technology in the identification and typing of microorganisms. This study employed the whole-cell MALDI-TOF MS to identify and differentiate wild-type and mutants containing constructed single gene mutations of Burkholderia pseudomallei, a pathogenic bacterium causing melioidosis disease in both humans and animals. Candidate biomarkers for the B. pseudomallei mutants, including rpoS, ppk, and bpsI isolates, were determined. Taxon-specific and clinical isolate-specific biomarkers of B. pseudomallei were consistently found and conserved across all average mass spectra. Cluster analysis of MALDI spectra of all isolates exhibited separate distribution. A total of twelve potential mass peaks discriminating between wild-type and mutant isolates were identified using ClinProTools analysis. Two peaks (m/z 2721 and 2748 Da) were specific for the rpoS isolate, three (m/z 3150, 3378, and 7994 Da) for ppk, and seven (m/z 3420, 3520, 3587, 3688, 4623, 4708, and 5450 Da) for bpsI. Our findings demonstrated that the rapid, accurate, and reproducible mass profiling technology could have new implications in laboratory-based rapid differentiation of extensive libraries of genetically altered bacteria. PMID:26656930

  15. Mitogen-Activated Protein Kinase Phosphorylation of Splicing Factor 45 (SPF45) Regulates SPF45 Alternative Splicing Site Utilization, Proliferation, and Cell Adhesion

    PubMed Central

    Al-Ayoubi, Adnan M.; Zheng, Hui; Liu, Yuying; Bai, Tao

    2012-01-01

    The regulation of alternative mRNA splicing factors by extracellular cues and signal transduction cascades is poorly understood. Using an engineered extracellular signal-regulated kinase 2 (ERK2) that can utilize ATP analogs, we have identified the alternative mRNA splicing factor 45 (SPF45), which is overexpressed in cancer, as a novel coimmunoprecipitating ERK2 substrate. ERK2 phosphorylated SPF45 on Thr71 and Ser222 in vitro and in cells in response to H-RasV12, B-RAF-V600E, and activated MEK1. Jun N-terminal kinase 1 (JNK1) and p38α also phosphorylated SPF45 in vitro and associated with SPF45 in cells. SPF45 was differentially phosphorylated in cells by all three mitogen-activated protein (MAP) kinases in response to phorbol myristate acid (PMA), H2O2, UV, and anisomycin stimulation. ERK and p38 activation decreased SPF45-dependent exon 6 exclusion from fas mRNA in a minigene assay in cells. Stable overexpression of SPF45 in SKOV-3 cells dramatically inhibited cell proliferation in a phosphorylation-dependent manner through inhibition of ErbB2 expression. SPF45 overexpression also induced EDA inclusion into fibronectin transcripts and fibronectin expression in a phosphorylation-dependent and -independent manner, respectively, specifically affecting cellular adhesion to a fibronectin matrix. These data identify SPF45 as the first splicing factor regulated by multiple MAP kinase pathways and show effects of both SPF45 overexpression and phosphorylation. PMID:22615491

  16. Ultrastructural analysis of central serotoninergic neurons immunolabeled by silver-gold-intensified diaminobenzidine chromogen. Completion of immunocytochemistry with X-ray microanalysis

    SciTech Connect

    Liposits, Z.; Goercs, T.T.; Trombitas, K.

    1985-06-01

    Serotonin immunoreactive structures of the rat central nervous system (CNS) were detected by the recently developed silver-intensified peroxidase-antiperoxidase complex (SI-PAP) method at both the light and electron microscopic levels. The silver postintensification of the diaminobenzidine (DAB) chromogen increased the sensitivity of the original PAP method, resulting in a very Golgi-like appearance of serotonin-immunopositive neuronal elements. The metallic silver and gold deposited onto DAB-labeled organelles, filling out the whole immunoreactive neuron, assures the easy tracing of thin neuronal processes far from the cell body. At the ultrastructural level, metallic grains were seen over immunolabeled structures only, proving the specificity of the silver method. In neurons of the dorsal raphe nucleus, free ribosomes, endoplasmic reticulum, and granules (80-100 nm in diameter) were labeled. Immunoreactive, e.g., serotoninergic, dendrites were seen to receive afferent terminals. The increased electron density of the intensified immunolabel facilitates the ultrastructural recognition of even weakly labeled profiles, while its metallic components (Ag and Au) provide a base for X-ray analysis of the immunolabeled biological specimen.

  17. Delayed positive gastrointestinal bleeding studies with technetium-99m-red blood cells: Utility of a second injection

    SciTech Connect

    Jacobson, A.F. )

    1991-02-01

    Two patients studied with technetium-99m-labeled red blood cells (RBCs) for gastrointestinal bleeding had positive findings only on 24-hr delayed images, at which time the site of bleeding could not be ascertained. In each instance, when additional delayed images suggested that active bleeding was occurring, a second aliquot of RBCs was labeled and injected. Sites of active hemorrhage were identified following further imaging in both patients. When delayed GI bleeding images are positive, further views should be obtained to ascertain if the pattern of intraluminal activity changes. If renewed active hemorrhage is suspected, reinjection with a second dose of labeled RBCs may identify the bleeding site.

  18. miRNA in situ hybridization in circulating tumor cells - MishCTC

    PubMed Central

    Ortega, Francisco G.; Lorente, Jose A.; Garcia Puche, Jose L.; Ruiz, Maria P.; Sanchez-Martin, Rosario M.; de Miguel-Pérez, Diego; Diaz-Mochon, Juan J.; Serrano, Maria J.

    2015-01-01

    Circulating tumor cells (CTCs) must be phenotypically and genetically characterized before they can be utilized in clinical applications. Here, we present the first protocol for the detection of miRNAs in CTCs using in situ hybridization (ISH) combined with immunomagnetic selection based on cytokeratin (CK) expression and immunocytochemistry. Locked-Nucleic Acid (LNA) probes associated with an enzyme-labeled fluorescence (ELF) signal amplification approach were used to detect miRNA-21 in CTCs. This protocol was optimized using both epithelial tumor (MDA-MB468) and epithelial non-tumor (MCF-10A) cell lines, and miRNA-21 was selected as the target miRNA because of its known role as an onco-miRNA. Hematopoietic cells do not express miRNA-21; thus, miRNA-21 is an ideal marker for detecting CTCs. Peripheral blood samples were taken from 25 cancer patients and these samples were analyzed using our developed protocol. Of the 25 samples, 11 contained CTCs. For all 11 CTC-positive samples, the isolated CTCs expressed both CK and miRNA-21. Finally, the protocol was applied to monitor miRNA-21 expression in epithelial to mesenchymal transition (EMT)-induced MCF-7 cells, an epithelial tumor cell line. CK expression was lost in these cells, whereas miRNA-21 was still expressed, suggesting that miRNA-21 might be a good marker for detecting CTCs with an EMT phenotype. PMID:25777797

  19. Enhancing the efficiency of flexible dye-sensitized solar cells utilizing natural dye extracted from Azadirachta indica

    NASA Astrophysics Data System (ADS)

    Sahare, Sanjay; Veldurthi, Naresh; Singh, Ranbir; Swarnkar, A. K.; Salunkhe, Manauti; Bhave, Tejashree

    2015-10-01

    The natural dye extracted from Azadirechta indica (neem) was used as a sensitizer in flexible dye-sensitized solar cells (DSSCs). The fabricated DSSC exhibited open circuit voltage of 0.538 V with 2.81% power conversion efficiency (η) in back-illuminated mode which is higher than that reported in the literature. In order to understand the characteristics of DSSC, systematic study of solar cell component materials was carried out. Anatase TiO2 (30-40 nm) nanoparticles were synthesized by DC arc plasma method and deposited electrophoretically on a flexible titanium (Ti) substrate. A platinum-coated polyethylene terephthalate (PET) substrate was used as a counter electrode to construct flexible DSSC. The structural and optical behavior of neem-dye sensitized TiO2 thin film has been studied using x-ray diffraction, field emission scanning electron microscopy, high resolution transmission electron microscopy and UV-visible spectroscopy. We have observed that the neem dye gives a very good sensitization effect. In addition, the dye has good prospects as a low-cost and environmental friendly alternative to ruthenium-based sensitizers which are normally used in DSSCs.

  20. Reduction in promotor methylation utilizing EGCG (epigallocatechin-3-gallate) restores RXRα expression in human colon cancer cells.

    PubMed

    Morris, Jay; Moseley, Vondina R; Cabang, April B; Coleman, Katie; Wei, Wei; Garrett-Mayer, Elizabeth; Wargovich, Michael J

    2016-06-01

    Silencing of regulatory genes through hypermethylation of CpG islands is an important mechanism in tumorigenesis. In colon cancer, RXRα, an important dimerization partner with other nuclear transcription factors, is silenced through this mechanism. We previously found that colon tumors in ApcMin/+ mice had diminished levels of RXRα protein and expression levels of this gene were restored by treatment with a green tea intervention, due to reduced promoter methylation of RXRα. We hypothesized that CIMP+ cell lines, which epigenetically silence key regulatory genes would also evidence silencing of RXRα and EGCG treatment would restore its expression. We indeed found EGCG to restore RXRα activity levels in the human cell lines, in a dose dependent manner and reduced RXRα promoter methylation. EGCG induced methylation changes in several other colon cancer related genes but did not cause a decrease in global methylation. Numerous epidemiological reports have shown the benefits of green tea consumption in reducing colon cancer risk but to date no studies have shown that the risk reduction may be related to the epigenetic restoration by tea polyphenols. Our results show that EGCG modulates the reversal of gene silencing involved in colon carcinogenesis providing a possible avenue for colon cancer prevention and treatment. PMID:27167203

  1. Utility of Ochrobactrum anthropi YC152 in a Microbial Fuel Cell as an Early Warning Device for Hexavalent Chromium Determination

    PubMed Central

    Wang, Guey-Horng; Cheng, Chiu-Yu; Liu, Man-Hai; Chen, Tzu-Yu; Hsieh, Min-Chi; Chung, Ying-Chien

    2016-01-01

    Fast hexavalent chromium (Cr(VI)) determination is important for environmental risk and health-related considerations. We used a microbial fuel cell-based biosensor inoculated with a facultatively anaerobic, Cr(VI)-reducing, and exoelectrogenic Ochrobactrum anthropi YC152 to determine the Cr(VI) concentration in water. The results indicated that O. anthropi YC152 exhibited high adaptability to pH, temperature, salinity, and water quality under anaerobic conditions. The stable performance of the microbial fuel cell (MFC)-based biosensor indicated its potential as a reliable biosensor system. The MFC voltage decreased as the Cr(VI) concentration in the MFC increased. Two satisfactory linear relationships were observed between the Cr(VI) concentration and voltage output for various Cr(VI) concentration ranges (0.0125–0.3 mg/L and 0.3–5 mg/L). The MFC biosensor is a simple device that can accurately measure Cr(VI) concentrations in drinking water, groundwater, and electroplating wastewater in 45 min with low deviations (<10%). The use of the biosensor can help in preventing the violation of effluent regulations and the maximum allowable concentration of Cr(VI) in water. Thus, the developed MFC biosensor has potential as an early warning detection device for Cr(VI) determination even if O. anthropi YC152 is a possible opportunistic pathogen. PMID:27537887

  2. Reduction in promotor methylation utilizing EGCG (epigallocatechin-3-gallate) restores RXRα expression in human colon cancer cells

    PubMed Central

    Morris, Jay; Moseley, Vondina R.; Cabang, April B.; Coleman, Katie; Wei, Wei; Garrett-Mayer, Elizabeth; Wargovich, Michael J.

    2016-01-01

    Silencing of regulatory genes through hypermethylation of CpG islands is an important mechanism in tumorigenesis. In colon cancer, RXRα, an important dimerization partner with other nuclear transcription factors, is silenced through this mechanism. We previously found that colon tumors in ApcMin/+ mice had diminished levels of RXRα protein and expression levels of this gene were restored by treatment with a green tea intervention, due to reduced promoter methylation of RXRα. We hypothesized that CIMP+ cell lines, which epigenetically silence key regulatory genes would also evidence silencing of RXRα and EGCG treatment would restore its expression. We indeed found EGCG to restore RXRα activity levels in the human cell lines, in a dose dependent manner and reduced RXRα promoter methylation. EGCG induced methylation changes in several other colon cancer related genes but did not cause a decrease in global methylation. Numerous epidemiological reports have shown the benefits of green tea consumption in reducing colon cancer risk but to date no studies have shown that the risk reduction may be related to the epigenetic restoration by tea polyphenols. Our results show that EGCG modulates the reversal of gene silencing involved in colon carcinogenesis providing a possible avenue for colon cancer prevention and treatment. PMID:27167203

  3. Utility of Ochrobactrum anthropi YC152 in a Microbial Fuel Cell as an Early Warning Device for Hexavalent Chromium Determination.

    PubMed

    Wang, Guey-Horng; Cheng, Chiu-Yu; Liu, Man-Hai; Chen, Tzu-Yu; Hsieh, Min-Chi; Chung, Ying-Chien

    2016-01-01

    Fast hexavalent chromium (Cr(VI)) determination is important for environmental risk and health-related considerations. We used a microbial fuel cell-based biosensor inoculated with a facultatively anaerobic, Cr(VI)-reducing, and exoelectrogenic Ochrobactrum anthropi YC152 to determine the Cr(VI) concentration in water. The results indicated that O. anthropi YC152 exhibited high adaptability to pH, temperature, salinity, and water quality under anaerobic conditions. The stable performance of the microbial fuel cell (MFC)-based biosensor indicated its potential as a reliable biosensor system. The MFC voltage decreased as the Cr(VI) concentration in the MFC increased. Two satisfactory linear relationships were observed between the Cr(VI) concentration and voltage output for various Cr(VI) concentration ranges (0.0125-0.3 mg/L and 0.3-5 mg/L). The MFC biosensor is a simple device that can accurately measure Cr(VI) concentrations in drinking water, groundwater, and electroplating wastewater in 45 min with low deviations (<10%). The use of the biosensor can help in preventing the violation of effluent regulations and the maximum allowable concentration of Cr(VI) in water. Thus, the developed MFC biosensor has potential as an early warning detection device for Cr(VI) determination even if O. anthropi YC152 is a possible opportunistic pathogen. PMID:27537887

  4. Utility of Ochrobactrum anthropi YC152 in a Microbial Fuel Cell as an Early Warning Device for Hexavalent Chromium Determination.

    PubMed

    Wang, Guey-Horng; Cheng, Chiu-Yu; Liu, Man-Hai; Chen, Tzu-Yu; Hsieh, Min-Chi; Chung, Ying-Chien

    2016-01-01

    Fast hexavalent chromium (Cr(VI)) determination is important for environmental risk and health-related considerations. We used a microbial fuel cell-based biosensor inoculated with a facultatively anaerobic, Cr(VI)-reducing, and exoelectrogenic Ochrobactrum anthropi YC152 to determine the Cr(VI) concentration in water. The results indicated that O. anthropi YC152 exhibited high adaptability to pH, temperature, salinity, and water quality under anaerobic conditions. The stable performance of the microbial fuel cell (MFC)-based biosensor indicated its potential as a reliable biosensor system. The MFC voltage decreased as the Cr(VI) concentration in the MFC increased. Two satisfactory linear relationships were observed between the Cr(VI) concentration and voltage output for various Cr(VI) concentration ranges (0.0125-0.3 mg/L and 0.3-5 mg/L). The MFC biosensor is a simple device that can accurately measure Cr(VI) concentrations in drinking water, groundwater, and electroplating wastewater in 45 min with low deviations (<10%). The use of the biosensor can help in preventing the violation of effluent regulations and the maximum allowable concentration of Cr(VI) in water. Thus, the developed MFC biosensor has potential as an early warning detection device for Cr(VI) determination even if O. anthropi YC152 is a possible opportunistic pathogen.

  5. Assessing the potential clinical utility of transplantations of neural and mesenchymal stem cells for treating neurodegenerative diseases.

    PubMed

    Lescaudron, Laurent; Boyer, C; Bonnamain, Virginie; Fink, K D; Lévêque, X; Rossignol, J; Nerrière-Daguin, V; Malouet, A C; Lelan, F; Dey, N D; Michel-Monigadon, D; Lu, M; Neveu, I; von Hörsten, S; Naveilhan, P; Dunbar, G L

    2012-01-01

    Treatments for neurodegenerative diseases have little impact on the long-term patient health. However, cellular transplants of neuroblasts derived from the aborted embryonic brain tissue in animal models of neurodegenerative disorders and in patients have demonstrated survival and functionality in the brain. However, ethical and functional problems due to the use of this fetal tissue stopped most of the clinical trials. Therefore, new cell sources were needed, and scientists focused on neural (NSCs) and mesenchymal stem cells (MSCs). When transplanted in the brain of animals with Parkinson's or Huntington's disease, NSCs and MSCs were able to induce partial functional recovery by promoting neuroprotection and immunomodulation. MSCs are more readily accessible than NSCs due to sources such as the bone marrow. However, MSCs are not capable of differentiating into neurons in vivo where NSCs are. Thus, transplantation of NSCs and MSCs is interesting for brain regenerative medicine. In this chapter, we detail the methods for NSCs and MSCs isolation as well as the transplantation procedures used to treat rodent models of neurodegenerative damage.

  6. Design of a titering assay for lentiviral vectors utilizing direct extraction of DNA from transduced cells in microtiter plates

    PubMed Central

    Murphy, Michele E; Vin, Chintan D; Slough, Megan M; Gombotz, Wayne R; Kelley-Clarke, Brenna

    2016-01-01

    Using lentiviral vector products in clinical applications requires an accurate method for measuring transduction titer. For vectors lacking a marker gene, quantitative polymerase chain reaction is used to evaluate the number of vector DNA copies in transduced target cells, from which a transduction titer is calculated. Immune Design previously described an integration-deficient lentiviral vector pseudotyped with a modified Sindbis virus envelope for use in cancer immunotherapy (VP02, of the ZVex platform). Standard protocols for titering integration-competent lentiviral vectors employ commercial spin columns to purify vector DNA from transduced cells, but such columns are not optimized for isolation of extrachromosomal (nonintegrated) DNA. Here, we describe a 96-well transduction titer assay in which DNA extraction is performed in situ in the transduction plate, yielding quantitative recovery of extrachromosomal DNA. Vector titers measured by this method were higher than when commercial spin columns were used for DNA isolation. Evaluation of the method’s specificity, linear range, and precision demonstrate that it is suitable for use as a lot release assay to support clinical trials with VP02. Finally, the method is compatible with titering both integrating and nonintegrating lentiviral vectors, suggesting that it may be used to evaluate the transduction titer for any lentiviral vector. PMID:26942209

  7. Immunocytochemistry of the acellular slime mold Physarum polycephalum. III. Distribution of myosin and the actin-modulating protein (fragmin) in sandwiched plasmodia.

    PubMed

    Osborn, M; Weber, K; Naib-Majani, W; Hinssen, H; Stockem, W; Wohlfarth-Bottermann, K E

    1983-01-01

    The acellular slime mold Physarum forms very thin plasmodia when sandwiched between two agar sheets. After extraction with glycerol-containing buffers, suitable objects for immunofluorescence microscopy are obtained, and an analysis of the cytoskeletal and contractile system of Physarum becomes possible. Plasmodia were stained with antibodies against myosin and fragmin, a protein factor involved in actin filament length regulation. The microanatomy and topography of cellular structures containing these proteins were investigated at the light and electron microscopic levels. The patterns obtained with the two antibodies are closely related to those obtained with actin antibody [25]. In both cases the complex system of cytoplasmic fibrils is stained selectively. The fibrils form a more or less regular network in the advancing front zone with the fibrils being interconnected by focal nodes. In the posterior region of the plasmodium, where endoplasmic pathways and protoplasmic veins are differentiated, larger fibrils are detected, running obliquely or longitudinally to the veins. With both antibodies the fluorescent pattern of the fibrils is continuous without indications of periodic interruptions or striations, which would be expected in the case of sarcomere-like subunits. With anti-myosin unstained patches are frequently seen at or close to the nodes of the fibrillar network in the anterior region. The small lobopodia, which are rich in actin, are apparently not stained by the myosin antibody, a result similar to the situation in "ruffling edges¿ of cultured vertebrate cells. Electron microscopic investigations of antibody-labeled fibrils in embedded and sectioned plasmodia allow the identification of antibody molecules at specific sites along the fibrils with a different distribution pattern for each of the two antibodies. PMID:6339244

  8. The utility of lactate dehydrogenase in the follow up of patients with diffuse large B-cell lymphoma

    PubMed Central

    William, Basem Magdy; Bongu, Navneeth Rao; Bast, Martin; Bociek, Robert Gregory; Bierman, Philip Jay; Vose, Julie Marie; Armitage, James Olen

    2013-01-01

    Background Serum lactate dehydrogenase is a non-specific marker for lymphoma whose prognostic significance is well established for both indolent and aggressive lymphomas at the time of diagnosis. The performance characteristics of this enzyme in predicting relapse in patients with diffuse large B-cell lymphoma has not been well studied. Methods This study compared serum lactate dehydrogenase levels in 27 patients with diffuse large B-cell lymphoma who relapsed after sustaining a complete response versus 87 patients who did not relapse. For relapsed patients, the serum lactate dehydrogenase level at relapse was compared with the level three months before (considered baseline). For non-relapsed patients, the last two levels during follow-up were compared. For statistical analysis the T-test was used to compare differences in mean values between groups. The sensitivity, specificity, positive and negative predictive values for serum lactate dehydrogenase in detecting relapse compared to confirmatory imaging were calculated. Results At relapse, only 33% patients had increases in serum lactate dehydrogenase above the upper limit of normal. The mean increase was 1.2-fold above the upper limit of normal for relapsed vs. 0.83 for those who did not relapse (p-value = 0.59). The mean increase in serum lactate dehydrogenase, from baseline, was 1.1-fold in non-relapsed vs. 1.3 in relapsed patients (p-value = 0.3). The likelihood ratio of relapse was 4.65 for patients who had 1.5-fold increases in serum lactate dehydrogenase above baseline (p-value = 0.03). The sensitivity, specificity, positive and negative predictive values of 1.5-fold increases for detecting relapse, compared to clinical and imaging findings were 0.18, 0.95, 0.55, and 0.79, respectively. Conclusion A 1.5-fold increase in serum lactate dehydrogenase, over a period of 3 months, is associated with increased likelihood of relapse from diffuse large B-cell lymphoma. PMID:23904809

  9. Potential utilization of waste sweetpotato vines hydrolysate as a new source for single cell oils production by Trichosporon fermentans.

    PubMed

    Zhan, Jumei; Lin, Hui; Shen, Qi; Zhou, Qifa; Zhao, Yuhua

    2013-05-01

    The enzymatic hydrolysate of sweetpotato vines (SVH) characterized as an effective nutrients supplier with low nitrogen availability was firstly used as a substrate by Trichosporon fermentans for single cell oils (SCOs) production. Batch-fermentation experiments on various SVH based media suggested that co-fermentation of SVH and some high-sugar content substrates would be much more efficient and less-cost for SCOs production. A lipid yield of 9.6 g l(-1) with a lipid content of 35.6% was achieved on the SVH without any addition, while 27.6 and 17.7 g l(-1) lipid were respectively obtained on the fructose supplemented SVH media and the SVH mixed with acid treated wheat straw hydrolysate (WSH). The positive effect of SVH on the lipid production of T. fermentans was further demonstrated with a kinetic investigation revealing that SVH had a remarkable promoting effect on the biomass formation and the substrate uptake. PMID:22985824

  10. Wrinkled silica/titania nanoparticles with tunable interwrinkle distances for efficient utilization of photons in dye-sensitized solar cells

    PubMed Central

    Kang, Jin Soo; Lim, Joohyun; Rho, Won-Yeop; Kim, Jin; Moon, Doo-Sik; Jeong, Juwon; Jung, Dongwook; Choi, Jung-Woo; Lee, Jin-Kyu; Sung, Yung-Eun

    2016-01-01

    Efficient light harvesting is essential for the realization of high energy conversion efficiency in dye-sensitized solar cells (DSCs). State-of-the-art mesoporous TiO2 photoanodes fall short for collection of long-wavelength visible light photons, and thus there have been efforts on introduction of scattering nanoparticles. Herein, we report the synthesis of wrinkled silica/titania nanoparticles with tunable interwrinkle distances as scattering materials for enhanced light harvesting in DSCs. These particles with more than 20 times larger specific surface area (>400 m2/g) compared to the spherical scattering particles (<20 m2/g) of the similar sizes gave rise to the dye-loading amounts, causing significant improvements in photocurrent density and efficiency. Moreover, dependence of spectral scattering properties of wrinkled particles on interwrinkle distances, which was originated from difference in overall refractive indices, was observed. PMID:27488465

  11. Wrinkled silica/titania nanoparticles with tunable interwrinkle distances for efficient utilization of photons in dye-sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Kang, Jin Soo; Lim, Joohyun; Rho, Won-Yeop; Kim, Jin; Moon, Doo-Sik; Jeong, Juwon; Jung, Dongwook; Choi, Jung-Woo; Lee, Jin-Kyu; Sung, Yung-Eun

    2016-08-01

    Efficient light harvesting is essential for the realization of high energy conversion efficiency in dye-sensitized solar cells (DSCs). State-of-the-art mesoporous TiO2 photoanodes fall short for collection of long-wavelength visible light photons, and thus there have been efforts on introduction of scattering nanoparticles. Herein, we report the synthesis of wrinkled silica/titania nanoparticles with tunable interwrinkle distances as scattering materials for enhanced light harvesting in DSCs. These particles with more than 20 times larger specific surface area (>400 m2/g) compared to the spherical scattering particles (<20 m2/g) of the similar sizes gave rise to the dye-loading amounts, causing significant improvements in photocurrent density and efficiency. Moreover, dependence of spectral scattering properties of wrinkled particles on interwrinkle distances, which was originated from difference in overall refractive indices, was observed.

  12. Wrinkled silica/titania nanoparticles with tunable interwrinkle distances for efficient utilization of photons in dye-sensitized solar cells.

    PubMed

    Kang, Jin Soo; Lim, Joohyun; Rho, Won-Yeop; Kim, Jin; Moon, Doo-Sik; Jeong, Juwon; Jung, Dongwook; Choi, Jung-Woo; Lee, Jin-Kyu; Sung, Yung-Eun

    2016-01-01

    Efficient light harvesting is essential for the realization of high energy conversion efficiency in dye-sensitized solar cells (DSCs). State-of-the-art mesoporous TiO2 photoanodes fall short for collection of long-wavelength visible light photons, and thus there have been efforts on introduction of scattering nanoparticles. Herein, we report the synthesis of wrinkled silica/titania nanoparticles with tunable interwrinkle distances as scattering materials for enhanced light harvesting in DSCs. These particles with more than 20 times larger specific surface area (>400 m(2)/g) compared to the spherical scattering particles (<20 m(2)/g) of the similar sizes gave rise to the dye-loading amounts, causing significant improvements in photocurrent density and efficiency. Moreover, dependence of spectral scattering properties of wrinkled particles on interwrinkle distances, which was originated from difference in overall refractive indices, was observed. PMID:27488465

  13. Degradation of cell wall polysaccharides by combinations of carbohydrase enzymes and their effect on nutrient utilization and broiler chicken performance.

    PubMed

    Meng, X; Slominski, B A; Nyachoti, C M; Campbell, L D; Guenter, W

    2005-01-01

    In vitro incubation studies were carried out to determine if various carbohydrase preparations contained appropriate activities to target nonstarch polysaccharides (NSP) of wheat, soybean meal (SBM), canola meal, and peas. Triplicate samples (0.1 g) were incubated with a number of carbohydrase preparations (i.e., cellulase, pectinase, xylanase, glucanase, galactanase, and mannanase) or their combinations at 45 degrees C and pH 5.2. A more pronounced degradation of NSP was achieved when the enzyme preparations were used in concert. When compared with the control (nonenzyme treatment), the highest degree of NSP degradation reached was 37% for wheat, and 36, 26, and 28% for canola meal, SBM, and peas, respectively. Four enzyme combinations were studied further in a 2-wk (5 to 18 d of age) growth performance and nutrient digestibility trial with broiler chickens. All enzyme combinations were effective in improving (P < 0.05) weight gain, feed-to-gain ratio, AMEn, apparent ileal digestibilities of starch and protein, and apparent total tract digestibility of NSP in birds fed a wheat, wheat screening, SBM, canola meal, and peas-based diet. The most complex enzyme combination was found to be superior (P < 0.05) to others in improving ileal protein digestibility and feed-to-gain ratio. The effectiveness of this combination in elimination of the nutrient-encapsulating effect of cell walls was further evaluated in a balance study with adult roosters fed a conventionally ground full-fat canola seed. Enzyme addition increased (P < 0.05) NSP digestibility from 11.1 to 30.1%, which, in turn, resulted in a marked increase (P < 0.05) in TMEn value (4.176 vs 4.744 Mcal/kg) of the seed. It is evident from the present studies that the addition of an appropriate combination of carbohydrase enzymes to target cell wall polysaccharide structures could further improve enzyme efficacy in practical wheat, SBM, canola meal, and peas-based broiler diets.

  14. Prognostic Utility of Squamous Cell Carcinoma Antigen in Carcinoma of the Cervix: Association With Pre- and Posttreatment FDG-PET

    SciTech Connect

    Olsen, Jeffrey R.; Dehdashti, Farrokh; Siegel, Barry A.; Zighelboim, Israel; Grigsby, Perry W.; Schwarz, Julie K.

    2011-11-01

    Purpose: Squamous cell carcinoma antigen (SCC Ag) is a serum biomarker for squamous cell carcinoma (SCC) of the cervix. We investigated the prognostic significance of SCC Ag levels before and at the completion of chemoradiotherapy and compared these levels with the results of pre- and posttreatment positron emission tomography/computed tomography (PET/CT) using [{sup 18}F]fluorodeoxyglucose (FDG). Methods and Materials: The records of 63 women who underwent definitive chemoradiotherapy for SCC of the cervix were reviewed. SCC Ag levels were obtained before and at the completion of radiotherapy. Patients were divided into two groups on the basis of their pretreatment SCC Ag level (>30 ng/mL vs. {<=}30 ng/mL). Pre- and posttreatment FDG-PET/CT characteristics and progression-free survival (PFS) were analyzed according to SCC Ag groups. Results: Median follow-up was 12 months. Women with SCC Ag >30 ng/mL at diagnosis had more advanced lymph node disease on pretreatment FDG-PET/CT than those with SCC Ag {<=}30 ng/mL (p = .002). Women whose SCC Ag normalized at the completion of chemoradiotherapy were more likely to have a complete metabolic response on their 3-month posttreatment FDG-PET/CT than those whose SCC Ag did not normalize (p = .006). The 2-year PFS was 73% for patients with a SCC Ag level {<=}30 ng/mL at diagnosis compared with 0% for those with a SCC Ag level >30 ng/mL at diagnosis (p < .0001). The 2-year PFS was 62% for patients whose SCC Ag normalized at the completion of chemoradiotherapy compared with 0% for those whose SCC Ag did not normalize (p = .0004). Conclusion: Elevated SCC Ag at diagnosis and failure of the SCC Ag to normalize at the completion of treatment are associated with incomplete metabolic response and decreased PFS.

  15. Quantitative, real-time analysis of base excision repair activity in cell lysates utilizing lesion-specific molecular beacons.

    PubMed

    Svilar, David; Vens, Conchita; Sobol, Robert W

    2012-01-01

    We describe a method for the quantitative, real-time measurement of DNA glycosylase and AP endonuclease activities in cell nuclear lysates using base excision repair (BER) molecular beacons. The substrate (beacon) is comprised of a deoxyoligonucleotide containing a single base lesion with a 6-Carboxyfluorescein (6-FAM) moiety conjugated to the 5'end and a Dabcyl moiety conjugated to the 3' end of the oligonucleotide. The BER molecular beacon is 43 bases in length and the sequence is designed to promote the formation of a stem-loop structure with 13 nucleotides in the loop and 15 base pairs in the stem. When folded in this configuration the 6-FAM moiety is quenched by Dabcyl in a non-fluorescent manner via Förster Resonance Energy Transfer (FRET). The lesion is positioned such that following base lesion removal and strand scission the remaining 5 base oligonucleotide containing the 6-FAM moiety is released from the stem. Release and detachment from the quencher (Dabcyl) results in an increase of fluorescence that is proportionate to the level of DNA repair. By collecting multiple reads of the fluorescence values, real-time assessment of BER activity is possible. The use of standard quantitative real-time PCR instruments allows the simultaneous analysis of numerous samples. The design of these BER molecular beacons, with a single base lesion, is amenable to kinetic analyses, BER quantification and inhibitor validation and is adaptable for quantification of DNA Repair activity in tissue and tumor cell lysates or with purified proteins. The analysis of BER activity in tumor lysates or tissue aspirates using these molecular beacons may be applicable to functional biomarker measurements. Further, the analysis of BER activity with purified proteins using this quantitative assay provides a rapid, high-throughput method for the discovery and validation of BER inhibitors.

  16. Rapamycin-mediated inhibition of mammalian target of rapamycin in skeletal muscle cells reduces glucose utilization and increases fatty acid oxidation.

    PubMed

    Sipula, Ian J; Brown, Nicholas F; Perdomo, German

    2006-12-01

    The mammalian target of rapamycin (mTOR) is a serine/threonine kinase that plays an important role in cell growth and metabolism. mTOR has been postulated as a nutrient sensor, but its role in the regulation of fatty acid and glucose metabolism is poorly understood. For the first time, we show that mTOR inhibition in skeletal muscle cells has pronounced effects on intermediary metabolism. Rapamycin, a uniquely specific mTOR inhibitor with clinical applications, increased fatty acid oxidation by 60% accompanied by increased activities of carnitine palmitoyltransferases I and II, the former believed to be the primary intracellular regulatory enzyme of the fatty acid oxidation pathway. Furthermore, glucose transport capacity, glycogen synthesis, and glycolysis were reduced by approximately 40% under the same conditions. In addition, in the presence of rapamycin, hyperinsulinemic conditions (100 nmol/L insulin, 24 hours) were unable to suppress fatty acid oxidation in L6 myotubes. Rapamycin treatment also decreased baseline phosphorylation of mTOR residues S2448 and S2481 by 30% and almost completely abolished p70 S6 kinase phosphorylation. These results show that rapamycin causes a metabolic shift from glucose utilization to fatty acid oxidation in model muscle cells in the presence of nutrient abundance and underline the importance of mTOR as a key regulator in glucose and lipid metabolism. PMID:17142137

  17. Identification of small molecule agonists of human relaxin family receptor 1 (RXFP1) by utilizing a homogenous cell-based cAMP assay

    PubMed Central

    Chen, Catherine Z.; Southall, Noel; Xiao, Jingbo; Marugan, Juan J.; Ferrer, Marc; Hu, Xin; Jones, Raisa E.; Feng, Shu; Agoulnik, Irina U.

    2016-01-01

    The relaxin hormone is involved in a variety of biological functions including female reproduction and parturition, regulation of cardiovascular, renal, pulmonary, and hepatic functions. It regulates extracellular matrix remodeling, cell invasiveness, proliferation, differentiation, and overall tissue homeostasis. The G protein-coupled receptor (GPCR) RXFP1, relaxin family receptor 1, is a cognate relaxin receptor that mainly signals through cyclic AMP second messenger. While agonists of the receptor could have a wide range of pharmacological utility, up to date, there are no reported small molecule agonists for relaxin receptors. Here, we report the development of quantitative high-throughput platform for RXFP1 agonist screen based on homogenous cell-based HTRF cAMP assay technology. Two small molecules of similar structure were independently identified from a screen of more than 365,677 compounds. Neither compound showed activity in a counter screen with HEK293T cells transfected with an unrelated GPCR vasopressin 1b receptor. These small molecule agonists also demonstrated selectivity against the RXFP2 receptor, providing a basis for future medicinal chemistry optimization of selective relaxin receptor agonists. PMID:23212924

  18. Effects of Dietary Supplementation of Barodon, an Anionic Alkali Mineral Complex, on Growth Performance, Feed Utilization, Innate Immunity, Goblet Cell and Digestibility in Olive Flounder (Paralichthys olivaceus)

    PubMed Central

    Shin, Chang-Hoon; Cha, Ji-Hoon; Rahimnejad, Samad; Jeong, Joon-Bum; Yoo, Byung-Woo; Lee, Bo-Kyeun; Ahn, Hyung-Jin; Choi, Soo-Il; Choi, Yun-Jeong; Park, Yong-Ho; Kim, Jeong-Dae; Lee, Kyeong-Jun

    2014-01-01

    A 15-wk feeding trial was conducted to examine the supplemental effects of Barodon on growth performance, gastrointestinal histology, feed digestibility and innate immunity in olive founder. A basal commercial diet was used as a control and two other diets were prepared by spraying 0.1% or 0.2% of Barodon. Triplicate groups of fish (BW, 145 g) were fed one of the test diets to apparent satiation twice daily. At the end of the feeding trial, fish growth performance was not significantly affected by dietary treatments; however, feed utilization was significantly improved (linear and quadratic, p<0.05) by Barodon supplementation. Significantly higher (p<0.05) survival rates were obtained in fish fed Barodon containing diets. Hepatosomatic index increased significantly in Barodon treated groups. Also, the use of Barodon resulted in significant increase (linear and quadratic, p<0.05) of intestine length and number of goblet cells. Significantly higher (Quadratic, p<0.05) apparent digestibility coefficient of DM was obtained by supplementation of Barodon. Lysozyme and myeloperoxidase activities increased quadratically and linearly, respectively, in Barodon treated fish. Also, significantly higher (linear and quadratic, p<0.05) superoxide dismutase activity was found in Barodon fed fish. The findings in this study show that inclusion of Barodon in diets for olive flounder improves feed utilization and digestibility, and positively affects digestive tract histology and innate immunity. PMID:25049965

  19. Enhancing the power generation in microbial fuel cells with effective utilization of goethite recovered from mining mud as anodic catalyst.

    PubMed

    Jadhav, Dipak A; Ghadge, Anil N; Ghangrekar, Makarand M

    2015-09-01

    Catalytic effect of goethite recovered from iron-ore mining mud was studied in microbial fuel cells (MFCs). Characterization of material recovered from mining mud confirms the recovery of iron oxide as goethite. Heat treated goethite (550 °C) and untreated raw goethite were coated on stainless-steel anode of MFC-1 and MFC-2, respectively; whereas, unmodified stainless-steel anode was used in MFC-3 (control). Fivefold increment in power was obtained in MFC-1 (17.1 W/m(3) at 20 Ω) than MFC-3 (3.5 W/m(3)). MFC with raw goethite coated anode also showed enhanced power (11 W/m(3)). Higher Coulombic efficiency (34%) was achieved in MFC-1 than control MFC-3 (13%). Decrease in mass-transport losses and higher redox current during electrochemical analyses support improved electron transfer with the use of goethite on anode. Cheaper goethite coating kinetically accelerates the electron transfer between bacteria and anode, proving to be a novel approach for enhancing the electricity generation along with organic matter removal in MFC. PMID:25983229

  20. A techno-economic comparison of fuel processors utilizing diesel for solid oxide fuel cell auxiliary power units

    NASA Astrophysics Data System (ADS)

    Nehter, Pedro; Hansen, John Bøgild; Larsen, Peter Koch

    Ultra-low sulphur diesel (ULSD) is the preferred fuel for mobile auxiliary power units (APU). The commercial available technologies in the kW-range are combustion engine based gensets, achieving system efficiencies about 20%. Solid oxide fuel cells (SOFC) promise improvements with respect to efficiency and emission, particularly for the low power range. Fuel processing methods i.e., catalytic partial oxidation, autothermal reforming and steam reforming have been demonstrated to operate on diesel with various sulphur contents. The choice of fuel processing method strongly affects the SOFC's system efficiency and power density. This paper investigates the impact of fuel processing methods on the economical potential in SOFC APUs, taking variable and capital cost into account. Autonomous concepts without any external water supply are compared with anode recycle configurations. The cost of electricity is very sensitive on the choice of the O/C ratio and the temperature conditions of the fuel processor. A sensitivity analysis is applied to identify the most cost effective concept for different economic boundary conditions. The favourite concepts are discussed with respect to technical challenges and requirements operating in the presence of sulphur.

  1. Enhancing the power generation in microbial fuel cells with effective utilization of goethite recovered from mining mud as anodic catalyst.

    PubMed

    Jadhav, Dipak A; Ghadge, Anil N; Ghangrekar, Makarand M

    2015-09-01

    Catalytic effect of goethite recovered from iron-ore mining mud was studied in microbial fuel cells (MFCs). Characterization of material recovered from mining mud confirms the recovery of iron oxide as goethite. Heat treated goethite (550 °C) and untreated raw goethite were coated on stainless-steel anode of MFC-1 and MFC-2, respectively; whereas, unmodified stainless-steel anode was used in MFC-3 (control). Fivefold increment in power was obtained in MFC-1 (17.1 W/m(3) at 20 Ω) than MFC-3 (3.5 W/m(3)). MFC with raw goethite coated anode also showed enhanced power (11 W/m(3)). Higher Coulombic efficiency (34%) was achieved in MFC-1 than control MFC-3 (13%). Decrease in mass-transport losses and higher redox current during electrochemical analyses support improved electron transfer with the use of goethite on anode. Cheaper goethite coating kinetically accelerates the electron transfer between bacteria and anode, proving to be a novel approach for enhancing the electricity generation along with organic matter removal in MFC.

  2. Novel brachytherapy treatment planning system utilizing dose rate dependent average cell survival, CT-simulator, and dose-volume histogram

    SciTech Connect

    Mayer, R.; Fong, W.; Frankel, T.

    1995-12-31

    This report describes a new brachytherapy planning program that provides an evaluation of a given low or high dose rate treatment taking into account spatial dose heterogeneity and cell response to radiation. This brachytherapy scheme uses the images from a CT-Simulator (AcQSim, Picker International, Cleveland, Ohio) to simultaneously localize the seed positions and to axially scan the patient. This procedure helps to ensure accurate registration of the putative seed positions with the patient tissues and organs. The seed positions are determined by back-projecting positions of seeds or dummy seeds from the CT-Simulator setup scout images. Physicians delineate the tissues of interest on the axial slices. Dose is computed after assigning activity (low dose rate) of dwell times (high dose rate) to the Ir{sup 192} or I{sup 125} seed. The planar isodose distribution is superimposed onto axial cuts of the tissues and onto coronal or sagital views of the tissues following image reconstruction. Areal or volumetric calculations of the dose distribution within a given tissue are computed from the tissue outlines. The treatment plan computes (1) volume differential and cummulative dose histograms of the dose delivered to individual tissues, (2) the average, standard deviation, and coefficient of skewness of the dose distribution delivered to the individual tissues, (3) the average survival probability for a given radiation treatment.

  3. The Utility of Positron Emission Tomography in the Treatment Planning of Image-Guided Radiotherapy for Non-Small Cell Lung Cancer

    PubMed Central

    Chi, Alexander; Nguyen, Nam P.

    2014-01-01

    In the thorax, the extent of tumor may be more accurately defined with the addition of 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET) to computed tomography (CT). This led to the increased utility of FDG-PET or PET/CT in the treatment planning of radiotherapy for non-small cell lung cancer (NSCLC). The inclusion of FDG-PET information in target volume delineation not only improves tumor localization but also decreases the amount of normal tissue included in the planning target volume (PTV) in selected patients. Therefore, it has a critical role in image-guided radiotherapy (IGRT) for NSCLC. In this review, the impact of FDG-PET on target volume delineation in radiotherapy for NSCLC, which may increase the possibility of safe dose escalation with IGRT, the commonly used methods for tumor target volume delineation FDG-PET for NSCLC, and its impact on clinical outcome will be discussed. PMID:25340040

  4. Nanowire-based hierarchical tin oxide/zinc stannate hollow microspheres: Enhanced solar energy utilization efficiency for dye-sensitized solar cells and photocatalytic degradation of dyes

    NASA Astrophysics Data System (ADS)

    Li, Zhengdao; Zhou, Yong; Mao, Wutao; Zou, Zhigang

    2015-01-01

    Nanowire-based SnO2/Zn2SnO4 hollow microspheres are synthesized using a facile one-pot method for solar energy conversion and environment cleaning. The micrometer-sized hollow spheres possess a hierarchical structure with the shell consisting of nanowires. With the hybrid SnO2/Zn2SnO4 microspheres as photoanodes, the dye-sensitized solar cells (DSSCs) with an overall 4.72% photoconversion efficiency is obtained, nearly 240% improvement over the DSSCs that uses nanorod-based hierarchical SnO2 microspheres. The hybrid microspheres are also determined to be high-performance photocatalyst with a better recyclability for the photodegradation of dyes under simulated sunlight irradiation. These improvements of solar energy utilization are ascribed to the formation of the heterojunctions between SnO2 and Zn2SnO4 to enhance electron transport and charge-separation efficiencies.

  5. An energy-harvesting scheme utilizing Ga-rich CuIn(1-x)GaxSe2 quantum dots for dye-sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Lin, Chin-An; Huang, K. P.; Ho, S. T.; Huang, Mei-Wen; He-Hau, Jr.

    2012-09-01

    Ga-rich CuIn(1-x)GaxSe2 (CIGS) quantum dots (QDs) with a wide bandgap of 1.58 eV were utilized in dye-sensitized solar cells for energy harvesting. Ga-rich CIGS QDs at TiO2 photoanodes afford the recombination reduction and thus suppress the dark current, leading to the increase of short-circuit current from 14.47 to 15.27 mA.cm-2 and open-circuit voltage from 751 to 762 mV. This is due to well-adjusted conduction band minimum of Ga-rich CIGS QDs between that of TiO2 and excited state oxidation potential of N719, enhancing the photoelectron collection and suppressing electron back-transfer from TiO2 to oxidized redox species in the electrolyte.

  6. Utilization of collaborative practice agreements between physicians and pharmacists as a mechanism to increase capacity to care for hematopoietic stem cell transplant recipients.

    PubMed

    Merten, Julianna A; Shapiro, Jamie F; Gulbis, Alison M; Rao, Kamakshi V; Bubalo, Joseph; Lanum, Scott; Engemann, Ashley Morris; Shayani, Sepideh; Williams, Casey; Leather, Helen; Walsh-Chocolaad, Tracey

    2013-04-01

    Survival after hematopoietic stem cell transplantation (HSCT) has improved and the number of allogeneic HSCTs performed annually in the United States is expected to reach 10,000 by 2015. The National Marrow Donor Program created the System Capacity Initiative to formulate mechanisms to care for the growing number of HSCT recipients. One proposed method to increase capacity is utilization of pharmacists to manage drug therapy via collaborative practice agreements (CPAs). Pharmacists have managed drug therapy in oncology patients with CPAs for decades; however, there are limited HSCT centers that employ this practice. Engaging in collaborative practice and billing agreements with credentialed pharmacists to manage therapeutic drug monitoring, chronic medical conditions, and supportive care in HSCT recipients may be cost-effective and enable physicians to spend more time on new or more complex patients. The goal of this paper is to provide a framework for implementation of a CPA and address how it may improve HSCT program capacity. PMID:23419976

  7. 13C NMR studies of gluconeogenesis in rat liver cells: Utilization of labeled glycerol by cells from euthyroid and hyperthyroid rats

    PubMed Central

    Cohen, S. M.; Ogawa, S.; Shulman, R. G.

    1979-01-01

    The gluconeogenic pathway from [2-13C]glycerol and [1,3-13C]glycerol has been followed in suspensions of isolated rat hepatocytes at 25°C by 13C NMR at 90.5 MHz. The flow of label through the major pathway from glycerol to L-glycerol 3-phosphate and into glucose was followed in cells from control and triiodothyronine-treated rats. Treatment increased the rates of glucose formation and glycerol consumption 2-fold and decreased the αGP level to 40%. We calculate that ≈60% of the flux is through the mitochondrial glycerol phosphate dehydrogenase in cells from triiodothyronine-treated rats, compared with ≈15% in cells from the controls. Equal distribution of label between the trioses of glucose was obtained and, because the C3-C4 spin-spin coupling gives the distribution of labeled carbons in the same molecule, it was possible to measure the amount of triose from unlabeled fructose incorporated into the glucose labeled at carbons 1, 3, 4, and 6. About 10% of the hexoses had flowed through the pentose cycle and back into the hexose pathway in cells from fasted rats. From the distribution of label at glucose carbons not labeled via the major pathway and from the carbon spin-spin splitting patterns observed, we conclude that transketolase is reversible whereas transaldolase is essentially irreversible in the nonoxidative pentose branch. PMID:287001

  8. Utility and translatability of mathematical modeling, cell culture and small and large animal models in magnetic nanoparticle hyperthermia cancer treatment research

    NASA Astrophysics Data System (ADS)

    Hoopes, P. J.; Petryk, Alicia A.; Misra, Adwiteeya; Kastner, Elliot J.; Pearce, John A.; Ryan, Thomas P.

    2015-03-01

    For more than 50 years, hyperthermia-based cancer researchers have utilized mathematical models, cell culture studies and animal models to better understand, develop and validate potential new treatments. It has been, and remains, unclear how and to what degree these research techniques depend on, complement and, ultimately, translate accurately to a successful clinical treatment. In the past, when mathematical models have not proven accurate in a clinical treatment situation, the initiating quantitative scientists (engineers, mathematicians and physicists) have tended to believe the biomedical parameters provided to them were inaccurately determined or reported. In a similar manner, experienced biomedical scientists often tend to question the value of mathematical models and cell culture results since those data typically lack the level of biologic and medical variability and complexity that are essential to accurately study and predict complex diseases and subsequent treatments. Such quantitative and biomedical interdependence, variability, diversity and promise have never been greater than they are within magnetic nanoparticle hyperthermia cancer treatment. The use of hyperthermia to treat cancer is well studied and has utilized numerous delivery techniques, including microwaves, radio frequency, focused ultrasound, induction heating, infrared radiation, warmed perfusion liquids (combined with chemotherapy), and, recently, metallic nanoparticles (NP) activated by near infrared radiation (NIR) and alternating magnetic field (AMF) based platforms. The goal of this paper is to use proven concepts and current research to address the potential pathobiology, modeling and quantification of the effects of treatment as pertaining to the similarities and differences in energy delivered by known external delivery techniques and iron oxide nanoparticles.

  9. Utility of interim and end-of-treatment PET/CT in peripheral T-cell lymphomas: A review of 124 patients.

    PubMed

    El-Galaly, Tarec Christoffer; Pedersen, Martin Bjerregård; Hutchings, Martin; Mylam, Karen Juul; Madsen, Jakob; Gang, Anne Ortved; Bøgsted, Martin; de Nully Brown, Peter; Loft, Annika; Nielsen, Anne Lerberg; Hendel, Helle Westergreen; Iyer, Victor; Gormsen, Lars Christian

    2015-11-01

    According to the updated guidelines for imaging in lymphoma, 18F-FDG positron emission tomography/computed tomography (PET/CT) is recommended for staging and evaluation of treatment response in FDG-avid lymphomas. The purpose of the study was to evaluate the utility of PET/CT in nodal peripheral T-cell lymphomas (PTCL). Patients with newly diagnosed nodal PTCL (peripheral T-cell lymphoma NOS, anaplastic large-cell lymphoma, or angioimmunoblastic T-cell lymphoma) seen at five Danish hematology centers during the period 2006 to 2012 were included, if they had been pretherapeutically staged with PET/CT. Medical records were reviewed for baseline clinical and follow-up information. Staging, interim (I-PET), and end-of-treatment PET/CT (E-PET) studies were centrally reviewed, and reported using the Deauville 5-point score (DS). A total of 124 patients fulfilled the inclusion criteria. The median age was 58 years, and 88% received CHOP/CHOP-like therapy. Five years PFS and OS of the study population was 36.8% (95% CI 27.3-46.4) and 49.7% (95% CI 38.9-59.6), respectively. The presence of PET/CT-ascertained lung and/or liver involvement was associated with a worse outcome. The sensitivity of PET/CT for detecting biopsy-defined bone marrow involvement was only 18% (95% CI 4-43). An interim DS >3 was not prognostic for worse OS and PFS among CHOP/CHOP-like treated patients in uni- or multivariate analyses. A DS >3 after treatment predicted a worse prognosis. In conclusion, I-PET was not predictive of outcome in CHOP/CHOP-like treated PTCL patients when using the DS. Prospective studies are needed to determine the optimal use of PET/CT in PTCL including the role of quantitative PET/CT analysis.

  10. Metanephric adenoma: the utility of immunohistochemical and cytogenetic analyses in differential diagnosis, including solid variant papillary renal cell carcinoma and epithelial-predominant nephroblastoma.

    PubMed

    Kinney, Stephanie N; Eble, John N; Hes, Ondrej; Williamson, Sean R; Grignon, David J; Wang, Mingsheng; Zhang, Shaobo; Baldrige, Lee Ann; Martignoni, Guido; Brunelli, Matteo; Wang, Lisha; Comperat, Eva; Fan, Rong; Montironi, Rodolfo; MacLennan, Gregory T; Cheng, Liang

    2015-09-01

    Metanephric adenoma is a benign renal neoplasm that overlaps in morphology with the solid variant of papillary renal cell carcinoma and epithelial-predominant nephroblastoma. To aid in resolving this differential diagnosis, we investigated the utility of immunohistochemical and molecular analyses in distinguishing between these entities; the first study, to our knowledge, to use a combined approach in analyzing all three tumors. We analyzed 37 tumors originally diagnosed as metanephric adenomas (2 of which we reclassified as papillary renal cell carcinomas), 13 solid variant papillary renal cell carcinomas, and 20 epithelial-predominant nephroblastomas using a combination of immunohistochemistry and fluorescence in situ hybridization (FISH) assessing for trisomy of chromosomes 7 and 17 and loss of Y. Immunohistochemical staining was performed for CK7, AMACR, WT1, and CD57. The combination of CK7-, AMACR-, WT1+, and CD57+ was considered characteristic of metanephric adenoma. Most of the tumors originally diagnosed as metanephric adenomas (31/37) showed the expected staining pattern of metanephric adenoma (CK7-, AMACR-, WT1+, and CD57+). Of the six tumors with discordant immunophenotype, two tumors were reclassified as papillary renal cell carcinoma after cytogenetic workup. It is recommended that all adult cases histologically resembling metanephric adenoma have WT1, CD57, CK7, and AMACR immunohistochemical staining performed. If the staining pattern is characteristic for metanephric adenoma (CK7-, AMACR-, WT1+, and CD57+, including membranous staining), then no other diagnostic tests are indicated. However, if there is a different immunostaining pattern, then we recommend FISH analysis.

  11. Hereditary leiomyomatosis and renal cell carcinoma syndrome-associated renal cancer: recognition of the syndrome by pathologic features and the utility of detecting aberrant succination by immunohistochemistry.

    PubMed

    Chen, Ying-Bei; Brannon, A Rose; Toubaji, Antoun; Dudas, Maria E; Won, Helen H; Al-Ahmadie, Hikmat A; Fine, Samson W; Gopalan, Anuradha; Frizzell, Norma; Voss, Martin H; Russo, Paul; Berger, Michael F; Tickoo, Satish K; Reuter, Victor E

    2014-05-01

    Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) syndrome is an autosomal dominant disorder in which germline mutations of fumarate hydratase (FH) gene confer an increased risk of cutaneous and uterine leiomyomas and renal cancer. HLRCC-associated renal cancer is highly aggressive and frequently presents as a solitary mass. We reviewed the clinicopathologic features of 9 patients with renal tumors presenting as sporadic cases but who were later proven to have FH germline mutations. Histologically, all tumors showed mixed architectural patterns, with papillary as the dominant pattern in only 3 cases. Besides papillary, tubular, tubulopapillary, solid, and cystic elements, 6 of 9 tumors contained collecting duct carcinoma-like areas with infiltrating tubules, nests, or individual cells surrounded by desmoplastic stroma. Prominent tubulocystic carcinoma-like component and sarcomatoid differentiation were identified. Although all tumors exhibited the proposed hallmark of HLRCC (large eosinophilic nucleolus surrounded by a clear halo), this feature was often not uniformly present throughout the tumor. Prior studies have shown that a high level of fumarate accumulated in HLRCC tumor cells causes aberrant succination of cellular proteins by forming a stable chemical modification, S-(2-succino)-cysteine (2SC), which can be detected by immunohistochemistry. We thus explored the utility of detecting 2SC by immunohistochemistry in the differential diagnosis of HLRCC tumors and other high-grade renal tumors and investigated the correlation between 2SC staining and FH molecular alterations. All confirmed HLRCC tumors demonstrated diffuse and strong nuclear and cytoplasmic 2SC staining, whereas all clear cell (184/184, 100%), most high-grade unclassified (93/97, 96%), and the large majority of "type 2" papillary (35/45, 78%) renal cell carcinoma cases showed no 2SC immunoreactivity. A subset of papillary (22%) and rare unclassified (4%) tumors showed patchy or diffuse

  12. Utility of endobronchial ultrasound-guided-fine-needle aspiration and additional value of cell block in the diagnosis of mediastinal granulomatous lymphadenopathy

    PubMed Central

    Zaidi, Shaesta Naseem; Raddaoui, Emad

    2015-01-01

    Background: Endobronchial ultrasound-guided transbronchial fine-needle aspiration is a minimally invasive technique for diagnosis of mediastinal lesions. Although most studies have reported the utility of EBUS-FNA in malignancy, its use has been extended to the benign conditions as well. Objective: To evaluate the diagnostic yield and cytologic accuracy of endobronchial ultrasound-guided transbronchial fine-needle aspiration (EBUS-FNA) in cases of clinically and radiologically suspected granulomatous diseases. Patients and Method: From May 2010 to April 2015, 43 of 115 patients who underwent EBUS-FNA at one center for radiologically and clinically suspicious granulomatous lesions, and with no definite histological diagnosis, were included in this retrospective study. Results: When the histological diagnosis was taken as the gold standard, the sensitivity of EBUS-FNA was 85% and specificity was 100% with the positive predictive value of 100. The combined diagnostic sensitivity of EBUS-FNA and transbronchial lung biopsy was 100%. In 4 cases, cell block provided an exclusive morphological diagnosis of sarcoidosis which was noncontributory by EBUS-FNA. Conclusion: Our study supports the use of EBUS-FNA, by virtue of being a safe, minimally invasive, and an outpatient procedure, in the diagnosis of granulomatous mediastinal lymphadenopathy, thereby obviating more invasive testing in a significant number of patients. Also, cell block provides additional data in the diagnosis in these benign mediastinal diseases. PMID:26445590

  13. The potential utility of acetyltanshinone IIA in the treatment of HER2-overexpressed breast cancer: Induction of cancer cell death by targeting apoptotic and metabolic signaling pathways

    PubMed Central

    Guerram, Mounia; Jiang, Zhen-Zhou; Yousef, Bashir Alsiddig; Hamdi, Aida Mejda; Hassan, Hozeifa Mohamed; Yuan, Zi-Qiao; Luo, Hou-Wei; Zhu, Xiong; Zhang, Lu-Yong

    2015-01-01

    Increased lipogenesis and protein synthesis is a hallmark of cancer cell proliferation, survival, and metastatic progression and is under intense investigation as a potential antineoplastic target. Acetyltanshinone IIA (ATA) is a compound that was obtained from chemical modifications of tanshinone IIA (TIIA), a potent anticancer agent extracted from the dried roots of the Chinese herbal medicine Salvia miltiorrhiza Bunge. A previous investigation indicated that ATA is more effective in inhibiting the growth of breast cancer especially cells with HER2 overexpression. However, the molecular mechanism(s) mediating this cytotoxic effect on HER2-positive breast cancer remained undefined. Studies described here report that ATA induced G1/S phase arrest and apoptosis in the HER2-positive MDA-MB-453, SK-BR-3, and BT-474 breast cancer cell lines. Mechanistic investigations revealed that the ATA-induced apoptosis effect is associated with remarkably down-regulation of receptor tyrosine kinases (RTKs) EGFR/HER2 and inhibition of their downstream pro-survival signaling pathways. Interestingly, ATA was found to trigger oxidative and endoplasmic reticulum (ER) stresses and to activate AMP activated protein kinase (AMPK) leading to inactivation of key enzymes involved in lipid and protein biogenesis. Intraperitoneal administration of ATA significantly inhibited the growth of MDA-MB-453 xenografts in athymic mice without causing weight loss and any other side effects. Additionally, transwell migration, invasion, and wound healing assays revealed that ATA could suppress tumor angiogenesis in vitro. Taken together, our data suggest that ATA may have broad utility in the treatment of HER2-overexpressed breast cancers. PMID:26068969

  14. An algorithm for utilizing peripheral blood CD34 count as a predictor of the need for plerixafor in autologous stem cell mobilization--cost-effectiveness analysis.

    PubMed

    Abusin, Ghada A; Abu-Arja, Rolla F; Gingrich, Roger D; Silverman, Margarida D; Zamba, Gideon K D; Schlueter, Annette J

    2013-08-01

    Certain patients who receive granulocyte colony-stimulating factor (GCSF) for autologous hematopoietic stem cell (AHSC) collection fail to mobilize well enough to proceed with transplant. When plerixafor is used with GCSF, the likelihood of achieving the CD34⁺ stem cell target in fewer collections is higher; plerixafor use in all patients is unlikely to be cost-effective. This study retrospectively evaluated the effectiveness of utilizing a peripheral blood CD34⁺ stem cell count (PBCD34) ≤8/µL on day 4 of GCSF-based AHSC mobilization as a threshold for plerixafor administration, and compared the efficacy of collection and cost analysis using historical controls. All patients in the study cohort reached their CD34⁺ targets in ≤3 collections. Significantly more patients who received plerixafor + GCSF versus GCSF alone reached their CD34⁺ target in one collection (P = 0.045); however, there were no significant differences in the number of collections or in cumulative product yields. The historical cohort had 10.3% mobilization failures; the number of collections per patient needed to reach the target was significantly higher in the historical cohort versus study cohort (P = 0.001) as was the number of patients requiring more than one collection to reach their target (P = 0.023). However, the average cost per patient was also significantly higher in the study cohort (P = 0.025). Further refinement of the algorithm may reduce the difference in cost between the two mobilization strategies.

  15. Prospective blinded study of somatic mutation detection in cell-free DNA utilizing a targeted 54-gene next generation sequencing panel in metastatic solid tumor patients.

    PubMed

    Kim, Seung Tae; Lee, Won-Suk; Lanman, Richard B; Mortimer, Stefanie; Zill, Oliver A; Kim, Kyoung-Mee; Jang, Kee Taek; Kim, Seok-Hyung; Park, Se Hoon; Park, Joon Oh; Park, Young Suk; Lim, Ho Yeong; Eltoukhy, Helmy; Kang, Won Ki; Lee, Woo Yong; Kim, Hee-Cheol; Park, Keunchil; Lee, Jeeyun; Talasaz, AmirAli

    2015-11-24

    Sequencing of the mutant allele fraction of circulating cell-free DNA (cfDNA) derived from tumors is increasingly utilized to detect actionable genomic alterations in cancer. We conducted a prospective blinded study of a comprehensive cfDNA sequencing panel with 54 cancer genes. To evaluate the concordance between cfDNA and tumor DNA (tDNA), sequencing results were compared between cfDNA from plasma and genomic tumor DNA (tDNA). Utilizing next generation digital sequencing technology (DST), we profiled approximately 78,000 bases encoding 512 complete exons in the targeted genes in cfDNA from plasma. Seventy-five patients were prospectively enrolled between February 2013 and March 2014, including 61 metastatic cancer patients and 14 clinical stage II CRC patients with matched plasma and tissue samples. Using the 54-gene panel, we detected at least one somatic mutation in 44 of 61 tDNA (72.1%) and 29 of 44 (65.9%) cfDNA. The overall concordance rate of cfDNA to tDNA was 85.9%, when all detected mutations were considered. We collected serial cfDNAs during cetuximab-based treatment in 2 metastatic KRAS wild-type CRC patients, one with acquired resistance and one with primary resistance. We demonstrate newly emerged KRAS mutation in cfDNA 1.5 months before radiologic progression. Another patient had a newly emerged PIK3CA H1047R mutation on cfDNA analysis at progression during cetuximab/irinotecan chemotherapy with gradual increase in allele frequency from 0.8 to 2.1%. This blinded, prospective study of a cfDNA sequencing showed high concordance to tDNA suggesting that the DST approach may be used as a non-invasive biopsy-free alternative to conventional sequencing using tumor biopsy.

  16. Biogas utilization

    SciTech Connect

    Moser, M.A.

    1995-11-01

    Options for successfully using biomass depend on project scale. Almost all biogas from anaerobic digesters must first go through a gas handling system that pressurizes, meters, and filters the biogas. Additional treatment, including hydrogen sulfide-mercaptan scrubbing, gas drying, and carbon dioxide removal may be necessary for specialized uses, but these are complex and expensive processes. Thus, they can be justified only for large-scale projects that require high-quality biogas. Small-scale projects (less than 65 cfm) generally use biogas (as produced) as a boiler fuel or for fueling internal combustion engine generators to produce electricity. If engines or boilers as selected properly, there should be no need to remove hydrogen sulfide. Small-scale combustion turbines, steam turbines, and fuel cells are not used because of their technical complexity and high capital cost. Biogas cleanup to pipeline or transportation fuel specification is very costly, and energy economics preclude this level of treatment.

  17. Biogas utilization

    SciTech Connect

    Moser, M.A.

    1996-01-01

    Options for successfully using biogas depend on project scale. Almost all biogas from anaerobic digesters must first go through a gas handling system that pressurizes, meters, and filters the biogas. Additional treatment, including hydrogen sulfide-mercaptan scrubbing, gas drying, and carbon dioxide removal may be necessary for specialized uses, but these are complex and expensive processes. Thus, they can be justified only for large-scale projects that require high-quality biogas. Small-scale projects (less than 65 cfm) generally use biogas (as produced) as a boiler fuel or for fueling internal combustion engine-generators to produce electricity. If engines or boilers are selected properly, there should be no need to remove hydrogen sulfide. Small-scale combustion turbines, steam turbines, and fuel cells are not used because of their technical complexity and high capital cost. Biogas cleanup to pipeline or transportation fuel specifications is very costly, and energy economics preclude this level of treatment.

  18. Life-cycle cost comparisons of advanced storage batteries and fuel cells for utility, stand-alone, and electric vehicle applications

    SciTech Connect

    Humphreys, K.K.; Brown, D.R.

    1990-01-01

    This report presents a comparison of battery and fuel cell economics for ten different technologies. To develop an equitable economic comparison, the technologies were evaluated on a life-cycle cost (LCC) basis. The LCC comparison involved normalizing source estimates to a standard set of assumptions and preparing a lifetime cost scenario for each technology, including the initial capital cost, replacement costs, operating and maintenance (O M) costs, auxiliary energy costs, costs due to system inefficiencies, the cost of energy stored, and salvage costs or credits. By considering all the costs associated with each technology over its respective lifetime, the technology that is most economical to operate over any given period of time can be determined. An analysis of this type indicates whether paying a high initial capital cost for a technology with low O M costs is more or less economical on a lifetime basis than purchasing a technology with a low initial capital cost and high O M costs. It is important to realize that while minimizing cost is important, the customer will not always purchase the least expensive technology. The customer may identify benefits associated with a more expensive option that make it the more attractive over all (e.g., reduced construction lead times, modularity, environmental benefits, spinning reserve, etc.). The LCC estimates presented in this report represent three end-use applications: utility load-leveling, stand-alone power systems, and electric vehicles.

  19. Optimization of the Energy Level Alignment between the Photoactive Layer and the Cathode Contact Utilizing Solution-Processed Hafnium Acetylacetonate as Buffer Layer for Efficient Polymer Solar Cells.

    PubMed

    Yu, Lu; Li, Qiuxiang; Shi, Zhenzhen; Liu, Hao; Wang, Yaping; Wang, Fuzhi; Zhang, Bing; Dai, Songyuan; Lin, Jun; Tan, Zhan'ao

    2016-01-13

    The insertion of an appropriate interfacial buffer layer between the photoactive layer and the contact electrodes makes a great impact on the performance of polymer solar cells (PSCs). Ideal interfacial buffer layers could minimize the interfacial traps and the interfacial barriers caused by the incompatibility between the photoactive layer and the electrodes. In this work, we utilized solution-processed hafnium(IV) acetylacetonate (Hf(acac)4) as an effective cathode buffer layer (CBL) in PSCs to optimize the energy level alignment between the photoactive layer and the cathode contact, with the short-circuit current density (Jsc), open-circuit voltage (Voc), and fill factor (FF) all simultaneously improved with Hf(acac)4 CBL, leading to enhanced power conversion efficiencies (PCEs). Ultraviolet photoemission spectroscopy (UPS) and scanning Kelvin probe microscopy (SKPM) were performed to confirm that the interfacial dipoles were formed with the same orientation direction as the built-in potential between the photoactive layer and Hf(acac)4 CBL, benefiting the exciton separation and electron transport/extraction. In addition, the optical characteristics and surface morphology of the Hf(acac)4 CBL were also investigated.

  20. Energetic-environmental-economic assessment of the biogas system with three utilization pathways: Combined heat and power, biomethane and fuel cell.

    PubMed

    Wu, Bin; Zhang, Xiangping; Shang, Dawei; Bao, Di; Zhang, Suojiang; Zheng, Tao

    2016-08-01

    A typical biogas system with three utilization pathways, i.e., biogas upgrading, biogas combined heat and power (CHP), biogas solid oxide fuel cells (SOFCs) were designed. It was assessed from the viewpoint of energy, environment and economy by using energy efficiency, green degree and net present value index respectively. The assessment considered the trade-off relationships among these indexes, which is more comprehensive than previous systematic evaluation work only included single or two of the pathway(s) by using one or two of the index(es). Assessment results indicated that biogas upgrading pathway has the highest systematic energy efficiency (46.5%) and shortest payback period (8.9year) with the green degree production is the lowest (9.29gd/day). While for biogas SOFC pathway, although the green degree production is the highest (21.77gd/day), the payback period is longer (14.5year) and the energy efficiency is 13.6% lower than the biogas upgrading pathway. PMID:27209454

  1. Sugar Utilization and Acid Production by Free and Entrapped Cells of Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis subsp. lactis in a Whey Permeate Medium

    PubMed Central

    Audet, Pascal; Paquin, Celine; Lacroix, Christophe

    1989-01-01

    Cells of Streptococcus salivarius subsp. thermophilus and Lactococcus lactis subsp. lactis entrapped in k-carrageenan-locust bean gum gel performed similarly to free cells in the conversion of lactose to lactic acid. Bead diameter influenced the fermentation rate. Cells entrapped in smaller beads (0.5 to 1.0 mm) showed higher release rates, higher lactose, glucose, and formic acid utilization, higher galactose accumulation, and higher lactic acid production than did cells entrapped in larger beads (1.0 to 2.0 mm). Values for smaller beads were comparable with those for free cells. Immobilization affected the fermentation rate of lactic acid bacteria, especially Lactobacillus delbrueckii subsp. bulgaricus. Entrapped cells of L. delbrueckii subsp. bulgaricus demonstrated a lower lactic acid production than did free cells in batch fermentation. The kinetics of the production of formic and pyruvic acids by L. lactis subsp. lactis and S. salivarius subsp. thermophilus are presented. PMID:16347822

  2. Human amniotic epithelial cells induce localized cell-mediated immune privilege in vitro: implications for pancreatic islet transplantation.

    PubMed

    Qureshi, Khalid M; Oliver, Robert J; Paget, Michelle B; Murray, Hilary E; Bailey, Clifford J; Downing, Richard

    2011-01-01

    Chronic systemic immunosuppression in cell replacement therapy restricts its clinical application. This study sought to explore the potential of cell-based immune modulation as an alternative to immunosuppressive drug therapy in the context of pancreatic islet transplantation. Human amniotic epithelial cells (AEC) possess innate anti-inflammatory and immunosuppressive properties that were utilized to create localized immune privilege in an in vitro islet cell culture system. Cellular constructs composed of human islets and AEC (islet/AEC) were bioengineered under defined rotational cell culture conditions. Insulin secretory capacity was validated by glucose challenge and immunomodulatory potential characterized using a peripheral blood lymphocyte (PBL) proliferation assay. Results were compared to control constructs composed of islets or AEC cultured alone. Studies employing AEC-conditioned medium examined the role of soluble factors, and fluorescence immunocytochemistry was used to identify putative mediators of the immunosuppressive response in isolated AEC monocultures. Sustained, physiologically appropriate insulin secretion was observed in both islets and islet/AEC constructs. Activation of resting PBL proliferation occurred on exposure to human islets alone but this response was significantly (p < 0.05) attenuated by the presence of AEC and AEC-conditioned medium. Mitogen (phytohaemagglutinin, 5 μg/ml)-induced PBL proliferation was sustained on contact with isolated islets but abrogated by AEC, conditioned medium, and the islet/AEC constructs. Immunocytochemical analysis of AEC monocultures identified a subpopulation of cells that expressed the proapoptosis protein Fas ligand. This study demonstrates that human islet/AEC constructs exhibit localized immunosuppressive properties with no impairment of β-cell function. The data suggest that transplanted islets may benefit from the immune privilege status conferred on them as a consequence of their close

  3. Myrtucommulone-A treatment decreases pluripotency- and multipotency-associated marker expression in bladder cancer cell line HTB-9.

    PubMed

    Iskender, Banu; Izgi, Kenan; Karaca, Halit; Canatan, Halit

    2015-10-01

    Cancer and stem cells exhibit similar features, including self-renewal, differentiation and immortality. The expression of stem-cell-related genes in cancer cells is demonstrated to be potentially correlated with cancer cell behaviour, affecting both drug response and tumor recurrence. There is an emerging body of evidence that subpopulations of tumors carry a distinct molecular sign and are selectively resistant to chemotherapy. Therefore, it is important to find novel therapeutic agents that could suppress the stem-like features of cancer cells while inhibiting their proliferation. Myrtucommulone-A (MC-A) is an active compound of a nonprenylated acylphloroglucinol isolated from the leaves of myrtle. Here we have investigated the potential of MC-A in inhibiting the expression of self-renewal regulatory factors and cancer stem cell markers in a bladder cancer cell line HTB-9. We used RT-PCR, immunocytochemistry, flow cytometry and western blotting to examine the expression of pluripotency- and multipotency-associated markers with or without treatment with MC-A. Treatment with MC-A not only decreased cancer cell viability and proliferation but also resulted in a decrease in the expression of pluripotency- and multipotency-associated markers such as NANOG, OCT-4, SOX-2, SSEA-4, TRA-1-60, CD90, CD73 and CD44. MC-A treatment was also observed to decrease the sphere-forming ability of HTB-9 cells. In summary, this study provides valuable information on the presence of stem-cell marker expression in HTB-9 cells and our results imply that MC-A could be utilized to target cancer cells with stem-like characteristics. PMID:26054707

  4. Clinical Genotyping of Non-Small Cell Lung Cancers Using Targeted Next-Generation Sequencing: Utility of Identifying Rare and Co-mutations in Oncogenic Driver Genes.

    PubMed

    Tafe, Laura J; Pierce, Kirsten J; Peterson, Jason D; de Abreu, Francine; Memoli, Vincent A; Black, Candice C; Pettus, Jason R; Marotti, Jonathan D; Gutmann, Edward J; Liu, Xiaoying; Shirai, Keisuke; Dragnev, Konstantin H; Amos, Christopher I; Tsongalis, Gregory J

    2016-09-01

    Detection of somatic mutations in non-small cell lung cancers (NSCLCs), especially adenocarcinomas, is important for directing patient care when targeted therapy is available. Here, we present our experience with genotyping NSCLC using the Ion Torrent Personal Genome Machine (PGM) and the AmpliSeq Cancer Hotspot Panel v2. We tested 453 NSCLC samples from 407 individual patients using the 50 gene AmpliSeq Cancer Hotspot Panel v2 from May 2013 to July 2015. Using 10 ng of DNA, up to 11 samples were simultaneously sequenced on the Ion Torrent PGM (316 and 318 chips). We identified variants with the Ion Torrent Variant Caller Plugin, and Golden Helix's SVS software was used for annotation and prediction of the significance of the variants. Three hundred ninety-eight samples were successfully sequenced (12.1% failure rate). In all, 633 variants in 41 genes were detected with a median of 2 (range of 0 to 7) variants per sample. Mutations detected in BRAF, EGFR, ERBB2, KRAS, NRAS, and PIK3CA were considered potentially actionable and were identified in 237 samples, most commonly in KRAS (37.9%), EGFR (11.1%), BRAF (4.8%), and PIK3CA (4.3%). In our patient population, all mutations in EGFR, KRAS, and BRAF were mutually exclusive. The Ion Torrent Ampliseq technology can be utilized on small biopsy and cytology specimens, requires very little input DNA, and can be applied in clinical laboratories for genotyping of NSCLC. This targeted next-generation sequencing approach allows for detection of common and also rare mutations that are clinically actionable in multiple patients simultaneously. PMID:27659017

  5. Proteinase production in Pseudomonas fluorescens ON2 is affected by carbon sources and allows surface-attached but not planktonic cells to utilize protein for growth in lake water.

    PubMed

    Nicolaisen, Mette H; Worm, Jakob; Jørgensen, Niels O G; Middelboe, Mathias; Nybroe, Ole

    2012-04-01

    Proteins may be an important carbon and nitrogen source to bacteria in aquatic habitats, yet knowledge on the actual utilization of this substrate by proteolytic bacteria is scarce. In this study, Pseudomonas fluorescens ON2 produced an alkaline proteinase (AprX) during growth, and there was no evidence for cell density-regulated or starvation-induced proteinase production. Proteinase was produced in the absence of an organic nitrogen source, and citrate had a negative while glucose had a positive effect on the production. Hence, P. fluorescens ON2 seems to exploit protein sources by expressing the proteinase during growth unless a preferred carbon source such as citrate is present. Lake water model systems were subsequently used to investigate the ability of proteolytic vs. nonproteolytic ON2 strains to utilize protein for growth at moderate cell densities. Only cells forming surface-attached microcolonies were able to utilize this resource, while planktonic cells were not. Our experiments are the first to experimentally support models predicting that production of extracellular enzymes in dilute environments may be a waste of resources, whereas it represents a favorable feeding strategy in organic matrices such as detritus, microcolonies, or biofilm. PMID:22224410

  6. Reinforced chitosan-based heart valve scaffold and utility of bone marrow-derived mesenchymal stem cells for cardiovascular tissue engineering

    NASA Astrophysics Data System (ADS)

    Albanna, Mohammad Zaki

    Recent research has demonstrated a strong correlation between the differentiation profile of mesenchymal stem cells (MSCs) and scaffold stiffness. Chitosan is being widely studied for tissue engineering applications due to its biocompatibility and biodegradability. However, its use in load-bearing applications is limited due to moderate to low mechanical properties. In this study, we investigated the effectiveness of a fiber reinforcement method for enhancing the mechanical properties of chitosan scaffolds. Chitosan fibers were fabricated using a solution extrusion and neutralization method and incorporated into porous chitosan scaffolds. The effects of different fiber/scaffold mass ratios, fiber mechanical properties and fiber lengths on scaffold mechanical properties were studied. The results showed that incorporating fibers improved scaffold strength and stiffness in proportion to the fiber/scaffold mass ratio. A fiber-reinforced heart valve leaflet scaffold achieved strength values comparable to the radial values of human pulmonary and aortic valves. Additionally, the effects of shorter fibers (2 mm) were found to be up to 3-fold greater than longer fibers (10 mm). Despite this reduction in fiber mechanical properties caused by heparin crosslinking, the heparin-modified fibers still improved the mechanical properties of the reinforced scaffolds, but to a lesser extent than the unmodified fibers. The results demonstrate that chitosan fiber-reinforcement can be used to generate tissue-matching mechanical properties in porous chitosan scaffolds and that fiber length and mechanical properties are important parameters in defining the degree of mechanical improvement. We further studied various chemical and physical treatments to improve the mechanical properties of chitosan fibers. With combination of chemical and physical treatments, fiber stiffness improved 40fold compared to unmodified fibers. We also isolated ovine bone marrow-derived MSCs and evaluated their

  7. Clinical utility of chimerism status assessed by lineage-specific short tandem repeat analysis: experience from four cases of allogeneic stem cell transplantation.

    PubMed

    Goh, Ri-Young; Cho, Sung-Suk; Song, Yoo-Jeong; Heo, Kyeong; Oh, Sung-Yong; Kim, Sung-Hyun; Kwon, Hyeok-Chan; Kim, Hyo-Jin; Han, Jin-Yeong

    2009-08-01

    Chimerism testing permits early prediction and documentation of successful engraftment, and also facilitates detection of impending graft rejection. In this study, we serially monitored chimerism status by short tandem repeat-based PCR in nucleated cells (NC), T cells and natural killer (NK) cells after myeloablative allogeneic stem cell transplantation (SCT). Four patients with myeloid malignancies showed discrepant chimerism results among those three fractions. Three patients had mixed chimerism (MC) of donor/host T cells at a time point around the onset of chronic graft-versus-host disease (GVHD). In two patients with disease relapse, MC of NK cells preceded a morphological relapse or NK cells showed a higher percentage of patient cells compared to NC. Therefore, our study shows that chimerism analysis in lineage-specific cells might be useful in predicting clinical outcome after allogeneic SCT in certain patients.

  8. 40 CFR Appendix A to Part 76 - Phase I Affected Coal-Fired Utility Units With Group 1 or Cell Burner Boilers

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Units With Group 1 or Cell Burner Boilers A Appendix A to Part 76 Protection of Environment... 1 or Cell Burner Boilers Table 1—Phase I Tangentially Fired Units State Plant Unit Operator ALABAMA... Vertically fired boiler. 2 Arch-fired boiler. Table 3—Phase I Cell Burner Technology Units State Plant...

  9. 40 CFR Appendix A to Part 76 - Phase I Affected Coal-Fired Utility Units With Group 1 or Cell Burner Boilers

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Units With Group 1 or Cell Burner Boilers A Appendix A to Part 76 Protection of Environment... 1 or Cell Burner Boilers Table 1—Phase I Tangentially Fired Units State Plant Unit Operator ALABAMA... Vertically fired boiler. 2 Arch-fired boiler. Table 3—Phase I Cell Burner Technology Units State Plant...

  10. 40 CFR Appendix A to Part 76 - Phase I Affected Coal-Fired Utility Units With Group 1 or Cell Burner Boilers

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Units With Group 1 or Cell Burner Boilers A Appendix A to Part 76 Protection of Environment... 1 or Cell Burner Boilers Table 1—Phase I Tangentially Fired Units State Plant Unit Operator ALABAMA... Vertically fired boiler. 2 Arch-fired boiler. Table 3—Phase I Cell Burner Technology Units State Plant...

  11. 40 CFR Appendix A to Part 76 - Phase I Affected Coal-Fired Utility Units With Group 1 or Cell Burner Boilers

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Units With Group 1 or Cell Burner Boilers A Appendix A to Part 76 Protection of Environment... 1 or Cell Burner Boilers Table 1—Phase I Tangentially Fired Units State Plant Unit Operator ALABAMA... Vertically fired boiler. 2 Arch-fired boiler. Table 3—Phase I Cell Burner Technology Units State Plant...

  12. 40 CFR Appendix A to Part 76 - Phase I Affected Coal-Fired Utility Units With Group 1 or Cell Burner Boilers

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Units With Group 1 or Cell Burner Boilers A Appendix A to Part 76 Protection of Environment... 1 or Cell Burner Boilers Table 1—Phase I Tangentially Fired Units State Plant Unit Operator ALABAMA... Vertically fired boiler. 2 Arch-fired boiler. Table 3—Phase I Cell Burner Technology Units State Plant...

  13. TFPI1 Mediates Resistance to Doxorubicin in Breast Cancer Cells by Inducing a Hypoxic-Like Response

    PubMed Central

    Davies, Gerald F.; Berg, Arnie; Postnikoff, Spike D. L.; Wilson, Heather L.; Arnason, Terra G.; Kusalik, Anthony; Harkness, Troy A. A.

    2014-01-01

    Thrombin and hypoxia are important players in breast cancer progression. Breast cancers often develop drug resistance, but mechanisms linking thrombin and hypoxia to drug resistance remain unresolved. Our studies using Doxorubicin (DOX) resistant MCF7 breast cancer cells reveals a mechanism linking DOX exposure with hypoxic induction of DOX resistance. Global expression changes between parental and DOX resistant MCF7 cells were examined. Westerns, Northerns and immunocytochemistry were used to validate drug resistance and differentially expressed genes. A cluster of genes involved in the anticoagulation pathway, with Tissue Factor Pathway Inhibitor 1 (TFPI1) the top hit, was identified. Plasmids overexpressing TFPI1 were utilized, and 1% O2 was used to test the effects of hypoxia on drug resistance. Lastly, microarray datasets from patients with drug resistant breast tumors were interrogated for TFPI1 expression levels. TFPI1 protein levels were found elevated in 3 additional DOX resistant cells lines, from humans and rats, indicating evolutionarily conservation of the effect. Elevated TFPI1 in DOX resistant cells was active, as thrombin protein levels were coincidentally low. We observed elevated HIF1α protein in DOX resistant cells, and in cells with forced expression of TFPI1, suggesting TFPI1 induces HIF1α. TFPI1 also induced c-MYC, c-SRC, and HDAC2 protein, as well as DOX resistance in parental cells. Growth of cells in 1% O2 induced elevated HIF1α, BCRP and MDR-1 protein, and these cells were resistant to DOX. Our in vitro results were consistent with in vivo patient datasets, as tumors harboring increased BCRP and MDR-1 expression also had increased TFPI1 expression. Our observations are clinically relevant indicating that DOX treatment induces an anticoagulation cascade, leading to inhibition of thrombin and the expression of HIF1α. This in turn activates a pathway leading to drug resistance. PMID:24489651

  14. A novel method to study insect olfactory receptor function using HEK293 cells.

    PubMed

    Corcoran, Jacob A; Jordan, Melissa D; Carraher, Colm; Newcomb, Richard D

    2014-11-01

    The development of rapid and reliable assays to characterize insect odorant receptors (ORs) and pheromone receptors (PRs) remains a challenge for the field. Typically ORs and PRs are functionally characterized either in vivo in transgenic Drosophila or in vitro through expression in Xenopus oocytes. While these approaches have succeeded, they are not well suited for high-throughput screening campaigns, primarily due to inherent characteristics that limit their ability to screen large quantities of compounds in a short period of time. The development of a practical, robust and consistent in vitro assay for functional studies on ORs and PRs would allow for high-throughput screening for ligands, as well as for compounds that could be used as novel olfactory-based pest management tools. Here we describe a novel method of utilizing human embryonic kidney cells (HEK293) transfected with inducible receptor constructs for the functional characterization of ORs in 96-well plates using a fluorescent spectrophotometer. Using EposOrco and EposOR3 from the pest moth, Epiphyas postvittana as an example, we generated HEK293 cell lines with robust and consistent responses to ligands in functional assays. Single-cell sorting of cell lines by FACS facilitated the selection of isogenic cell lines with maximal responses, and the addition of epitope tags on the N-termini allowed the detection of recombinant proteins in homogenates by western blot and in cells by immunocytochemistry. We thoroughly describe the methods used to generate these OR-expressing cell lines, demonstrating that they have all the necessary features required for use in high-throughput screening platforms.

  15. In vitro differentiation potential of human haematopoietic CD34(+) cells towards pancreatic β-cells.

    PubMed

    Sunitha, Manne Mudhu; Srikanth, Lokanathan; Santhosh Kumar, Pasupuleti; Chandrasekhar, Chodimella; Sarma, Potukuchi Venkata Gurunadha Krishna

    2016-10-01

    Haematopoietic stem cells (HSCs) possess multipotent ability to differentiate into various types of cells on providing appropriate niche. In the present study, the differentiating potential of human HSCs into β-cells of islets of langerhans was explored. Human HSCs were apheretically isolated from a donor and cultured. Phenotypic characterization of CD34 glycoprotein in the growing monolayer HSCs was confirmed by immunocytochemistry and flow cytometry techniques. HSCs were induced by selection with beta cell differentiating medium (BDM), which consists of epidermal growth factor (EGF), fibroblast growth factor (FGF), transferrin, Triiodo-l-Tyronine, nicotinamide and activin A. Distinct morphological changes of differentiated cells were observed on staining with dithizone (DTZ) and expression of PDX1, insulin and synaptophysin was confirmed by immunocytochemistry. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed distinct expression of specific β-cell markers, pancreatic and duodenal homeobox-1 (PDX1), glucose transporter-2 (GLUT-2), synaptophysin (SYP) and insulin (INS) in these differentiated cells compared to HSCs. Further, these cells exhibited elevated expression of INS gene at 10 mM glucose upon inducing with different glucose concentrations. The prominent feature of the obtained β-cells was the presence of glucose sensors, which was determined by glucokinase activity and high glucokinase activity compared with CD34(+) stem cells. These findings illustrate the differentiation of CD34(+) HSCs into β-cells of islets of langerhans. PMID:27514733

  16. Lunar construction utility vehicle

    NASA Technical Reports Server (NTRS)

    1989-01-01

    The lunar construction utility vehicle (LCUV) is an all-purpose construction vehicle which will aid in the robotic assembly of a lunar outpost. The LCUV will have the following capabilities: (1) must be self supporting including repairs; (2) must offload itself from a lunar lander; (3) must be telerobotic and semi-autonomous; (4) must be able to transport one space station common module; (5) must allow for man-rated operation; and (6) must be able to move lunar regolith for site preparation. This study recommends the use of an elastic tracked vehicle. Detailed material analyses of most of the LCUV components were accomplished. The body frame, made of pinned truss elements, was stress analyzed using NASTRAN. A track connection system was developed; however, kinematic and stress analyses are still required. This design recommends the use of hydrogen-oxygen fuel cells for power. Thermal control has proven to be a problem which may be the most challenging technically. A tentative solution has been proposed which utilizes an onboard and towable radiator. Detailed study of the heat dissipation requirements is needed to finalize radiator sizing. Preliminary work on a man-rated cabin has begun; however, this is not required during the first mission phase of the LCUV. Finally, still in the conceptual phases, are the communication, navigation and mechanical arm systems.

  17. Selective utilization of Toll-like receptor and MyD88 signaling in B cells for enhancement of the anti-viral germinal center response

    PubMed Central

    Hou, Baidong; Saudan, Philippe; Ott, Gary; Wheeler, Matthew L.; Ji, Ming; Kuzmich, Lili; Lee, Linda M.; Coffman, Robert L.; Bachmann, Martin F.; DeFranco, Anthony L.

    2011-01-01

    Summary The contribution of Toll-like receptor (TLR) signaling to T cell-dependent (TD) antibody responses was assessed by using mice lacking the TLR signaling adaptor MyD88 in individual cell types. When a soluble TLR9 ligand was used as adjuvant for a protein antigen, MyD88 was required in dendritic cells but not in B cells to enhance the TD antibody response, regardless of the inherent immunogenicity of the antigen. In contrast, a TLR9 ligand contained within a virus-like particle substantially augmented the TD germinal center IgG antibody response, and this augmentation required B cell MyD88. The ability of B cells to discriminate between antigens based the physical form of a TLR ligand likely reflects an adaptation to facilitate strong anti-viral antibody responses. PMID:21353603

  18. Utilization of human DC-SIGN and L-SIGN for entry and infection of host cells by the New World arenavirus, Junín virus

    PubMed Central

    Belouzard, Sandrine; Cordo, Sandra M.; Candurra, Nélida A.; Whittaker, Gary R.

    2014-01-01

    The target cell tropism of enveloped viruses is regulated by interactions between viral proteins and cellular receptors determining susceptibility at a host cell, tissue or species level. However, a number of additional cell-surface moieties can also bind viral envelope glycoproteins and could act as capture receptors, serving as attachment factors to concentrate virus particles on the cell surface, or to disseminate the virus infection to target organs or susceptible cells within the host. Here, we used Junín virus (JUNV) or JUNV glycoprotein complex (GPC)-pseudotyped particles to study their ability to be internalized by the human C-type lectins hDC- or hL-SIGN. Our results provide evidence that hDC- and hL-SIGN can mediate the entry of Junín virus into cells, and may play an important role in virus infection and dissemination in the host. PMID:24183720

  19. Controlling Your Utility Rates.

    ERIC Educational Resources Information Center

    Lucht, Ray; Dembowski, Frederick L.

    1985-01-01

    A cost-effective alternative to high utility bills for middle-sized and smaller utility users is the service of utility rate consultants. The consultants analyze utility invoices for the previous 12 months to locate available refunds or credits. (MLF)

  20. Broader utilization of origins of DNA replication in cancer cell lines along a 78 kb region of human chromosome 2q34.

    PubMed

    Valenzuela, Manuel S; Hu, Lan; Lueders, John; Walker, Robert; Meltzer, Paul S

    2012-01-01

    Human DNA replication depends on the activation of thousands of origins distributed within the genome. The actual distribution of origins is not known, nor whether this distribution is unique to a cell type, or if it changes with the proliferative state of the cell. In this study, we have employed a real-time PCR-based nascent strand DNA abundance assay, to determine the location of origins along a 78 kb region on Chr2q34. Preliminary studies using nascent DNA strands isolated from either HeLa and normal skin fibroblast cells showed that in both cell lines peaks of high origin activity mapped in similar locations. However, the overall origin profile in HeLa cells corresponded to broad origin activation zones, whereas in fibroblasts a more punctuated profile of origin activation was observed. To investigate the relevance of this differential origin profile, we compared the origin distribution profiles in breast cancer cell lines MDA-MB-231, BT-474, and MCF-7, to their normal counterpart MCF-10A. In addition, the CRL7250 cell line was also used as a normal control. Our results validated our earlier observation and showed that the origin profile in normal cell lines exhibited a punctuated pattern, in contrast to broader zone profiles observed in the cancer cell lines. A quantitative analysis of origin peaks revealed that the number of activated origins in cancer cells is statistically larger than that obtained in normal cells, suggesting that the flexibility of origin usage is significantly increased in cancer cells compared to their normal counterparts.

  1. Establishment of a production-ready manufacturing process utilizing thin silicon substrates for solar cells. Final report. Motorola report No. 2364/4

    SciTech Connect

    Pryor, R. A.

    1980-10-01

    Three inch diameter Czochralski silicon substrates sliced directly to 5 mil, 8 mil, and 27 mil thicknesses with wire saw techniques were procured. Processing sequences incorporating either diffusion or ion implantation technologies were employed to produce n+p or n+pp+ solar cell structures. These cells were evaluated for performance, ease of fabrication, and cost effectiveness. It was determined that the use of 7 mil or even 4 mil wafers would provide near term cost reductions for solar cell manufacturers.

  2. Improving gene transfer in human renal carcinoma cells: Utilization of adenovirus vectors containing chimeric type 5 and type 35 fiber proteins

    PubMed Central

    ACHARYA, BISHNU; TERAO, SHUJI; SUZUKI, TORU; NAOE, MICHIO; HAMADA, KATSUYUKI; MIZUGUCHI, HIROYUKI; GOTOH, AKINOBU

    2010-01-01

    The transduction efficacy of adenovirus serotype 5 (Ad5) vector in human renal carcinoma cells is generally low due to the down-regulated expression of Coxsackie and adenovirus receptor (CAR) in target cells. By contrast, the infectivity of adenovirus serotype 35 vectors depends on the binding rate to CD46 receptor, independent of CAR. In this study, we examined whether an adenovirus vector containing chimeric type 5 and type 35 fiber proteins (Ad5/F35) increases transduction efficiency compared to Ad5 vector in human renal carcinoma cells in vitro. The expression of CAR was much lower in the human renal carcinoma cells than in control HEK293 cells. By contrast, the expression of CD46 was similar and perhaps at a higher level in the human renal carcinoma cells than in the HEK293 cells. The transduction efficacy of Ad5/F35 vector was dramatically higher compared to that of Ad5 in human renal carcinoma cells, and was correlated to the expression of CD46. Thus, Ad5/35 vector may be useful for the development of novel gene therapy approaches to renal cell carcinoma. PMID:22993573

  3. Improving gene transfer in human renal carcinoma cells: Utilization of adenovirus vectors containing chimeric type 5 and type 35 fiber proteins.

    PubMed

    Acharya, Bishnu; Terao, Shuji; Suzuki, Toru; Naoe, Michio; Hamada, Katsuyuki; Mizuguchi, Hiroyuki; Gotoh, Akinobu

    2010-05-01

    The transduction efficacy of adenovirus serotype 5 (Ad5) vector in human renal carcinoma cells is generally low due to the down-regulated expression of Coxsackie and adenovirus receptor (CAR) in target cells. By contrast, the infectivity of adenovirus serotype 35 vectors depends on the binding rate to CD46 receptor, independent of CAR. In this study, we examined whether an adenovirus vector containing chimeric type 5 and type 35 fiber proteins (Ad5/F35) increases transduction efficiency compared to Ad5 vector in human renal carcinoma cells in vitro. The expression of CAR was much lower in the human renal carcinoma cells than in control HEK293 cells. By contrast, the expression of CD46 was similar and perhaps at a higher level in the human renal carcinoma cells than in the HEK293 cells. The transduction efficacy of Ad5/F35 vector was dramatically higher compared to that of Ad5 in human renal carcinoma cells, and was correlated to the expression of CD46. Thus, Ad5/35 vector may be useful for the development of novel gene therapy approaches to renal cell carcinoma.

  4. Characterization of amniotic stem cells.

    PubMed

    Koike, Chika; Zhou, Kaixuan; Takeda, Yuji; Fathy, Moustafa; Okabe, Motonori; Yoshida, Toshiko; Nakamura, Yukio; Kato, Yukio; Nikaido, Toshio

    2014-08-01

    The amnion membrane is developed from embryo-derived cells, and amniotic cells have been shown to exhibit multidifferentiation potential. These cells represent a desirable source for stem cells for a variety of reasons. However, to date very few molecular analyses of amnion-derived cells have been reported, and efficient markers for isolating the stem cells remain unclear. This paper assesses the characterization of amnion-derived cells as stem cells by examining stemness marker expressions for amnion-derived epithelial cells and mesenchymal cells by flow cytometry, immunocytochemistry, and quantitative PCR. Flow cytometry revealed that amnion epithelial cells expressed CD133, CD 271, and TRA-1-60, whereas mecenchymal cells expressed CD44, CD73, CD90, and CD105. Immunohistochemistry showed that both cells expressed the stemness markers Oct3/4, Sox2, Klf4, and SSEA4. Stemness genes' expression in amnion epithelial cells, mesenchymal cells, fibroblast, bone marrow-derived mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs) was compared by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Amnion-derived epithelial cells and mesenchymal cells expressed Oct3/4, Nanog, and Klf4 more than bone marrow-derived MSCs. The sorted TRA1-60-positive cells expressed Oct3/4, Nanog, and Klf4 more than unsorted cells or TRA1-60-negative cells. TRA1-60 can be a marker for isolating amnion epithelial stem cells.

  5. Transplantation with cultured stem cells derived from the human amniotic membrane for corneal alkali burns: an experimental study.

    PubMed

    Zeng, Wei; Li, Yanwei; Zeng, Guangwei; Yang, Bo; Zhu, Yu

    2014-01-01

    Amniotic membranes (AM) have been used in a wide range of clinical applications. We successfully extracted mesenchymal stem cells (MSCs) from human AM, but little is known about the use and efficacy of human amniotic membrane-derived mesenchymal stem cells (hAM-dMSCs) for the treatment of alkali burns. We utilized hAM-dMSCs transplantation, AM grafting, and their combined use in the treatment of alkali burns. An experimental model in rabbits was devised to analyze the use of these techniques with immunocytochemistry and ELISA. The survival and migration of hAM-dMSCs labeled by SPION in the host were assessed with Prussian blue staining. Compared with the control group, the treated groups demonstrated faster reconstruction of the corneal epithelium, and lower levels of corneal opacification and neovascularization within corneal alkali burns. Furthermore, dark blue-stained particles were detected in the limbus corneae at day 28. These results demonstrated the ability of hAM-dMSCs to enhance epithelial healing and reduce corneal opacification and neovascularization in corneal alkali wounds.

  6. T-independent and T-dependent B lymphoblasts: helper T cells prime for interleukin 2-induced growth and secretion of immunoglobulins that utilize downstream heavy chains.

    PubMed

    Forman, M S; Puré, E

    1991-03-01

    Resting B cells enlarge, enter the cell cycle, and change their surface phenotype when activated via the surface immunoglobulin (Ig) receptor, but subsequent cell growth and antibody production is relatively limited. To identify stimuli that might prime B cells for enhanced function in vitro, we have compared the effects of anti-Ig with helper T (Th) cells on the formation of B lymphoblasts and the subsequent ability of the blasts to grow and secrete Ig. The B blasts first were induced by either anti-Ig, anti-Ig plus T cell-derived lymphokines, or alloreactive T blasts. Each population of B blasts showed enhanced expression of cell surface adhesion molecules, interleukin 2 receptor (IL-2R) p55, and MHC products, as well as decreased expression of IgD. The allo-activated B blasts were distinctive in expressing low levels of Thy-1 and increased reactivity with peanut agglutinin, a marker of germinal center B blasts in situ. The function of the different populations of B blasts was also different. Whereas anti-Ig or anti-Ig plus lymphokines primed for enhanced responses to lipopolysaccharide (LPS), the B blasts induced by Th cells were insensitive to LPS. B lymphoblasts that had been activated in the presence of helper factors or Th cells responded vigorously to recombinant IL-2 with growth and Ig secretion, and this response was enhanced in the presence of anti-Ig. The B blasts activated directly by Th cells, but not by anti-Ig plus lymphokines, were primed to secrete high levels of IgG1 and IgA. Therefore, the phenotype and function of a B lymphoblast depends upon the manner in which it is primed. When primed by Th cells, IL-2 proves to be the predominant mediator of clonal expansion and antibody secretion.

  7. T lymphocytes and mast cells express messenger RNA for interleukin-4 in the nasal mucosa in allergen-induced rhinitis.

    PubMed

    Ying, S; Durham, S R; Jacobson, M R; Rak, S; Masuyama, K; Lowhagen, O; Kay, A B; Hamid, Q A

    1994-06-01

    We have investigated the phenotype of interleukin-4 (IL-4) mRNA+ cells in the nasal mucosa of six subjects with allergic rhinitis before and 24 hr after local allergen provocation with grass pollen extract. Serial cryostat sections were cut from paraformaldehyde-fixed snap-frozen nasal biopsies, and immunocytochemistry (APAAP) followed by in situ hybridization performed on the same sections. For immunocytochemistry, antibodies against CD3, tryptase, major basic protein (MBP) and CD68 were used to identify T cells, mast cells, eosinophils and macrophages, respectively. Hybridization studies were performed using a digoxigenin-labelled IL-4 riboprobe. Nitroblue tetrazolium (NBT) and X-phosphate-5-bromo-4-chloro-3-indoly phosphate (BCIP) served as chromogens to detect hybridization IL-4 mRNA signals. Significant increases in T lymphocytes and eosinophils and in the number of IL-4 mRNA+ cells were observed after allergen challenge. Double immunocytochemistry/in situ hybridization demonstrated that the majority of IL-4 mRNA+ cells after allergen challenge were CD3+ (73.7% +/- 1.6). Lower numbers of IL-4 mRNA hybridization signals were co-localized to tryptase+ cells (26.0% +/- 1.6). In contrast, no IL-4 mRNA hybridization signals were co-localized to either eosinophils or macrophages. These results indicate that after allergen challenge T cells are the principal cellular source of IL-4 mRNA transcripts during human late nasal responses, with a lesser contribution from mast cells.

  8. Clinical utility of neuronal cells directly converted from fibroblasts of patients for neuropsychiatric disorders: studies of lysosomal storage diseases and channelopathy.

    PubMed

    Kano, S; Yuan, M; Cardarelli, R A; Maegawa, G; Higurashi, N; Gaval-Cruz, M; Wilson, A M; Tristan, C; Kondo, M A; Chen, Y; Koga, M; Obie, C; Ishizuka, K; Seshadri, S; Srivastava, R; Kato, T A; Horiuchi, Y; Sedlak, T W; Lee, Y; Rapoport, J L; Hirose, S; Okano, H; Valle, D; O'Donnell, P; Sawa, A; Kai, M

    2015-01-01

    Methodologies for generating functional neuronal cells directly from human fibroblasts [induced neuronal (iN) cells] have been recently developed, but the research so far has only focused on technical refinements or recapitulation of known pathological phenotypes. A critical question is whether this novel technology will contribute to elucidation of novel disease mechanisms or evaluation of therapeutic strategies. Here we have addressed this question by studying Tay-Sachs disease, a representative lysosomal storage disease, and Dravet syndrome, a form of severe myoclonic epilepsy in infancy, using human iN cells with feature of immature postmitotic glutamatergic neuronal cells. In Tay-Sachs disease, we have successfully characterized canonical neuronal pathology, massive accumulation of GM2 ganglioside, and demonstrated the suitability of this novel cell culture for future drug screening. In Dravet syndrome, we have identified a novel functional phenotype that was not suggested by studies of classical mouse models and human autopsied brains. Taken together, the present study demonstrates that human iN cells are useful for translational neuroscience research to explore novel disease mechanisms and evaluate therapeutic compounds. In the future, research using human iN cells with well-characterized genomic landscape can be integrated into multidisciplinary patient-oriented research on neuropsychiatric disorders to address novel disease mechanisms and evaluate therapeutic strategies.

  9. Clinical utility of neuronal cells directly converted from fibroblasts of patients for neuropsychiatric disorders: studies of lysosomal storage diseases and channelopathy.

    PubMed

    Kano, S; Yuan, M; Cardarelli, R A; Maegawa, G; Higurashi, N; Gaval-Cruz, M; Wilson, A M; Tristan, C; Kondo, M A; Chen, Y; Koga, M; Obie, C; Ishizuka, K; Seshadri, S; Srivastava, R; Kato, T A; Horiuchi, Y; Sedlak, T W; Lee, Y; Rapoport, J L; Hirose, S; Okano, H; Valle, D; O'Donnell, P; Sawa, A; Kai, M

    2015-01-01

    Methodologies for generating functional neuronal cells directly from human fibroblasts [induced neuronal (iN) cells] have been recently developed, but the research so far has only focused on technical refinements or recapitulation of known pathological phenotypes. A critical question is whether this novel technology will contribute to elucidation of novel disease mechanisms or evaluation of therapeutic strategies. Here we have addressed this question by studying Tay-Sachs disease, a representative lysosomal storage disease, and Dravet syndrome, a form of severe myoclonic epilepsy in infancy, using human iN cells with feature of immature postmitotic glutamatergic neuronal cells. In Tay-Sachs disease, we have successfully characterized canonical neuronal pathology, massive accumulation of GM2 ganglioside, and demonstrated the suitability of this novel cell culture for future drug screening. In Dravet syndrome, we have identified a novel functional phenotype that was not suggested by studies of classical mouse models and human autopsied brains. Taken together, the present study demonstrates that human iN cells are useful for translational neuroscience research to explore novel disease mechanisms and evaluate therapeutic compounds. In the future, research using human iN cells with well-characterized genomic landscape can be integrated into multidisciplinary patient-oriented research on neuropsychiatric disorders to address novel disease mechanisms and evaluate therapeutic strategies. PMID:25732146

  10. Directed Differentiation of Human Embryonic Stem Cells into Neural Progenitors.

    PubMed

    Banda, Erin; Grabel, Laura

    2016-01-01

    A variety of protocols have been used to produce neural progenitors from human embryonic stem cells. We have focused on a monolayer culture approach that generates neural rosettes. To initiate differentiation, cells are plated in a serum-free nutrient-poor medium in the presence of a BMP inhibitor. Depending on the cell line used, additional growth factor inhibitors may be required to promote neural differentiation. Long-term culture and addition of the Notch inhibitor DAPT can promote terminal neuronal differentiation. Extent of differentiation is monitored using immunocytochemistry for cell type-specific markers.

  11. Amacrine cells in the ganglion cell layer of the cat retina.

    PubMed

    Wässle, H; Chun, M H; Müller, F

    1987-11-15

    Following transection of the optic nerve, ganglion cells in the cat retina undergo retrograde degeneration. However, many small profiles (less than or equal to 10 micron) survive in the ganglion cell layer. Previously considered to be neuroglia, there is now substantial evidence that they are displaced amacrine cells. Their density increases from approximately 1,000 cells/mm2 in peripheral retina to 7,000 cells/mm2 in the central area. Their total number was found to be 850,000, which is five times the number of ganglion cells and also five times the number of astrocytes. Uptake of 3H-muscimol followed by autoradiography labelled 75% of the displaced amacrine cells; hence, the majority seem to be GABAergic. Immunocytochemistry with an antibody directed against choline-acetyl-transferase labelled approximately 10% of the displaced amacrines in the peripheral retina and 17% in the central area. Uptake of serotonin (5-HT) followed by immunocytochemistry was found in 25-30% of displaced amacrines. NADPH diaphorase histochemistry labelled approximately 5% of displaced amacrine cells. The sum of the various percentages make colocalization likely. Intracellular injection of Lucifer Yellow under microscopic control revealed that displaced amacrine cells constitute several morphological types. PMID:3693612

  12. Human endometrial stem cells as a new source for programming to neural cells.

    PubMed

    Mobarakeh, Zahra Taherian; Ai, Jafar; Yazdani, Farzad; Sorkhabadi, Seyed Mahdi Rezayat; Ghanbari, Zinat; Javidan, Abbas Noroozi; Mortazavi-Tabatabaei, Seyed Abdolreza; Massumi, Mohammad; Barough, Somayeh Ebrahimi

    2012-04-26

    Human EnSC (endometrial-derived stem cell) is an abundant and easily available source for cell replacement therapy. Many investigations have shown the potency of the cells to differentiate into several mesoderm-derived cell lineages, including osteocytes and adipocytes. Here, the potency of EnSC in neural differentiation has been investigated. Flow cytometric analysis showed that they were positive for CD90, CD105, OCT4, CD44 and negative for CD31, CD34, CD133. The characterized cells were induced into neural differentiation by bFGF (basic fibroblast growth factor), PDGF (platelet-derived growth factor) and EGF (epidermal growth factor) signalling molecules, respectively in a sequential protocol, and differentiated cells were analysed for expression of neuronal markers by RT-PCR (reverse transcription-PCR) and immunocytochemistry, including Nestin, GABA (γ-aminobutyric acid), MAP2 (microtubule-associated protein 2), β3-tub (class III β-tubulin) and NF-L (neurofilament-light) at the level of their mRNAs. The expression of MAP2, β3-tub and NF-L proteins in EnSC was confirmed 28 days PT (post-treatment) by immunocytochemistry. In conclusion, EnSC can respond to signalling molecules that are usually used as standards in neural differentiation and can programme neuronal cells, making these cells worth considering as a unique source for cell therapy in neurodegenerative disease.

  13. Metal insulator semiconductor solar cell devices based on a Cu{sub 2}O substrate utilizing h-BN as an insulating and passivating layer

    SciTech Connect

    Ergen, Onur; Gibb, Ashley; Vazquez-Mena, Oscar; Zettl, Alex; Regan, William Raymond

    2015-03-09

    We demonstrate cuprous oxide (Cu{sub 2}O) based metal insulator semiconductor Schottky (MIS-Schottky) solar cells with efficiency exceeding 3%. A unique direct growth technique is employed in the fabrication, and hexagonal boron nitride (h-BN) serves simultaneously as a passivation and insulation layer on the active Cu{sub 2}O layer. The devices are the most efficient of any Cu{sub 2}O based MIS-Schottky solar cells reported to date.

  14. Metal insulator semiconductor solar cell devices based on a Cu2O substrate utilizing h-BN as an insulating and passivating layer

    NASA Astrophysics Data System (ADS)

    Ergen, Onur; Gibb, Ashley; Vazquez-Mena, Oscar; Regan, William Raymond; Zettl, Alex

    2015-03-01

    We demonstrate cuprous oxide (Cu2O) based metal insulator semiconductor Schottky (MIS-Schottky) solar cells with efficiency exceeding 3%. A unique direct growth technique is employed in the fabrication, and hexagonal boron nitride (h-BN) serves simultaneously as a passivation and insulation layer on the active Cu2O layer. The devices are the most efficient of any Cu2O based MIS-Schottky solar cells reported to date.

  15. The clinical utility of PD-L1 testing in selecting non-small cell lung cancer patients for PD1/PD-L1-directed therapy.

    PubMed

    Villaruz, L C; Socinski, M A

    2016-09-01

    Lung cancer is the leading cause of cancer mortality in the United States and worldwide. Long thought to be nonimmunogenic, immunotherapy in lung cancer has historically been met with disappointing results. Programmed death-1 (PD-1), and the PD-1 ligand, PD-L1, are immune checkpoint proteins that fine-tune the antigen-specific T-cell response after stimulation of the T-cell receptor and are crucial for self-tolerance. This pathway in particular is co-opted by tumors through expression of PD-L1 on the tumor cell surface and within the tumor microenvironment, allowing for direct suppression of antitumor cytolytic T-cell activity by the tumor. Indeed, induction of the PD1/PD-L1 pathway represents an adaptive immune resistance mechanism exerted by tumor cells in response to endogenous antitumor activity. In 2015, the US Food and Drug Administration (FDA) approved two immuno-oncology agents, the PD-1 inhibitors nivolumab and pembrolizumab, for the treatment of previously treated advanced non-small cell lung cancer (NSCLC). Coincident with the clinical trials that led to these regulatory approvals has been the development of several immunohistochemistry (IHC) tests of PD-L1 expression, which may serve to select patients who will derive the most benefit from PD1- or PD-L1-directed therapy. The PD-L1 IHC assays are distinct in their methods and interpretation, which poses a challenge to clinicians selecting patients for these therapies.

  16. Fluorinated Fe(III) salophene complexes: optimization of tumor cell specific activity and utilization of fluorine labeling for in vitro analysis.

    PubMed

    Würtenberger, Irene; Follia, Valeria; Lerch, Fanni; Cwikla, Christiane; Fahrner, Nathalie; Kalchschmidt, Christina; Flögel, Brigitte; Kircher, Brigitte; Gust, Ronald

    2015-01-22

    Fluorine-substituted iron(III) salophene complexes (salophene = N,N'-bis(salicylidene)-1,2-phenylenediamine) were synthesized and evaluated for biological activity. All complexes showed growth inhibitory effects with IC50 values ranging from 0.05 to 2.45 μM against HT-29 colon carcinoma as well as MCF-7 and MDA-MB-231 mammary carcinoma cells (cisplatin: 5.75, 12.72, 5.81 μM, respectively). HR-CS MAS investigations revealed that the complexes were highly protein-bound already after an incubation period of 10 min and accumulated more effectively in tumor cells than cisplatin. Interestingly, the ligands were enriched in the cells too, indicating that the salophene moiety acts as a carrier ligand and mediates the uptake of the complexes. Furthermore, induction of apoptosis proved to be dependent on the substitution pattern as well as on the tumor cell line, as evidenced from the annexin V-FITC/PI assay. Most of the complexes, especially the highly active 5-Fe, showed tumor cell specific effects and no/less influence on the proliferation of T-cells generated from the peripheral blood of healthy individuals. PMID:25495038

  17. Fluorinated Fe(III) salophene complexes: optimization of tumor cell specific activity and utilization of fluorine labeling for in vitro analysis.

    PubMed

    Würtenberger, Irene; Follia, Valeria; Lerch, Fanni; Cwikla, Christiane; Fahrner, Nathalie; Kalchschmidt, Christina; Flögel, Brigitte; Kircher, Brigitte; Gust, Ronald

    2015-01-22

    Fluorine-substituted iron(III) salophene complexes (salophene = N,N'-bis(salicylidene)-1,2-phenylenediamine) were synthesized and evaluated for biological activity. All complexes showed growth inhibitory effects with IC50 values ranging from 0.05 to 2.45 μM against HT-29 colon carcinoma as well as MCF-7 and MDA-MB-231 mammary carcinoma cells (cisplatin: 5.75, 12.72, 5.81 μM, respectively). HR-CS MAS investigations revealed that the complexes were highly protein-bound already after an incubation period of 10 min and accumulated more effectively in tumor cells than cisplatin. Interestingly, the ligands were enriched in the cells too, indicating that the salophene moiety acts as a carrier ligand and mediates the uptake of the complexes. Furthermore, induction of apoptosis proved to be dependent on the substitution pattern as well as on the tumor cell line, as evidenced from the annexin V-FITC/PI assay. Most of the complexes, especially the highly active 5-Fe, showed tumor cell specific effects and no/less influence on the proliferation of T-cells generated from the peripheral blood of healthy individuals.

  18. Monitoring utilizations of amino acids and vitamins in culture media and Chinese hamster ovary cells by liquid chromatography tandem mass spectrometry.

    PubMed

    Qiu, Jinshu; Chan, Pik Kay; Bondarenko, Pavel V

    2016-01-01

    Monitoring amino acids and vitamins is important for understanding human health, food nutrition and the culture of mammalian cells used to produce therapeutic proteins in biotechnology. A method including ion pairing reversed-phase liquid chromatography with tandem mass spectrometry was developed and optimized to quantify 21 amino acids and 9 water-soluble vitamins in Chinese hamster ovary (CHO) cells and culture media. By optimizing the chromatographic separation, scan time, monitoring time window, and sample preparation procedure, and using isotopically labeled (13)C, (15)N and (2)H internal standards, low limits of quantitation (≤0.054 mg/L), good precision (<10%) and good accuracy (100±10%) were achieved for nearly all the 30 compounds. Applying this method to CHO cell extracts, statistically significant differences in the metabolite levels were measured between two cell lines originated from the same host, indicating differences in genetic makeup or metabolic activities and nutrient supply levels in the culture media. In a fed-batch process of manufacturing scale bioreactors, two distinguished trends for changes in amino acid concentrations were identified in response to feeding. Ten essential amino acids showed a zigzag pattern with maxima at the feeding days, and 9 non-essential amino acids displayed a smoothly changing profile as they were mainly products of cellular metabolism. Five of 9 vitamins accumulated continuously during the culture period, suggesting that they were fed in access. The method serves as an effective tool for the development and optimization of mammalian cell cultures. PMID:26355770

  19. Some personal and historical notes on the utility of "deep-etch" electron microscopy for making cell structure/function correlations.

    PubMed

    Heuser, John E

    2014-11-01

    This brief essay talks up the advantages of metal replicas for electron microscopy and explains why they are still the best way to image frozen cells in the electron microscope. Then it explains our approach to freezing, namely the Van Harreveld trick of "slamming" living cells onto a supercold block of metal sprayed with liquid helium at -269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment. This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most "true to life" views of cells out of them with a minimum of hassle. Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the "founding fathers" that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are "irresistible and half transparent … their meaning buried under only a few years of work," and "reasonable working hypotheses are already suggested by the ultrastructural organization itself."

  20. The Utilization of Plant Facilities on the International Space Station—The Composition, Growth, and Development of Plant Cell Walls under Microgravity Conditions

    PubMed Central

    Jost, Ann-Iren Kittang; Hoson, Takayuki; Iversen, Tor-Henning

    2015-01-01

    In the preparation for missions to Mars, basic knowledge of the mechanisms of growth and development of living plants under microgravity (micro-g) conditions is essential. Focus has centered on the g-effects on rigidity, including mechanisms of signal perception, transduction, and response in gravity resistance. These components of gravity resistance are linked to the evolution and acquisition of responses to various mechanical stresses. An overview is given both on the basic effect of hypergravity as well as of micro-g conditions in the cell wall changes. The review includes plant experiments in the US Space Shuttle and the effect of short space stays (8–14 days) on single cells (plant protoplasts). Regeneration of protoplasts is dependent on cortical microtubules to orient the nascent cellulose microfibrils in the cell wall. The space protoplast experiments demonstrated that the regeneration capacity of protoplasts was retarded. Two critical factors are the basis for longer space experiments: a. the effects of gravity on the molecular mechanisms for cell wall development, b. the availability of facilities and hardware for performing cell wall experiments in space and return of RNA/DNA back to the Earth. Linked to these aspects is a description of existing hardware functioning on the International Space Station. PMID:27135317

  1. The Utilization of Plant Facilities on the International Space Station-The Composition, Growth, and Development of Plant Cell Walls under Microgravity Conditions.

    PubMed

    Jost, Ann-Iren Kittang; Hoson, Takayuki; Iversen, Tor-Henning

    2015-01-20

    In the preparation for missions to Mars, basic knowledge of the mechanisms of growth and development of living plants under microgravity (micro-g) conditions is essential. Focus has centered on the g-effects on rigidity, including mechanisms of signal perception, transduction, and response in gravity resistance. These components of gravity resistance are linked to the evolution and acquisition of responses to various mechanical stresses. An overview is given both on the basic effect of hypergravity as well as of micro-g conditions in the cell wall changes. The review includes plant experiments in the US Space Shuttle and the effect of short space stays (8-14 days) on single cells (plant protoplasts). Regeneration of protoplasts is dependent on cortical microtubules to orient the nascent cellulose microfibrils in the cell wall. The space protoplast experiments demonstrated that the regeneration capacity of protoplasts was retarded. Two critical factors are the basis for longer space experiments: a. the effects of gravity on the molecular mechanisms for cell wall development, b. the availability of facilities and hardware for performing cell wall experiments in space and return of RNA/DNA back to the Earth. Linked to these aspects is a description of existing hardware functioning on the International Space Station.

  2. Some personal and historical notes on the utility of "deep-etch" electron microscopy for making cell structure/function correlations.

    PubMed

    Heuser, John E

    2014-11-01

    This brief essay talks up the advantages of metal replicas for electron microscopy and explains why they are still the best way to image frozen cells in the electron microscope. Then it explains our approach to freezing, namely the Van Harreveld trick of "slamming" living cells onto a supercold block of metal sprayed with liquid helium at -269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment. This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most "true to life" views of cells out of them with a minimum of hassle. Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the "founding fathers" that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are "irresistible and half transparent … their meaning buried under only a few years of work," and "reasonable working hypotheses are already suggested by the ultrastructural organization itself." PMID:25360049

  3. Monitoring utilizations of amino acids and vitamins in culture media and Chinese hamster ovary cells by liquid chromatography tandem mass spectrometry.

    PubMed

    Qiu, Jinshu; Chan, Pik Kay; Bondarenko, Pavel V

    2016-01-01

    Monitoring amino acids and vitamins is important for understanding human health, food nutrition and the culture of mammalian cells used to produce therapeutic proteins in biotechnology. A method including ion pairing reversed-phase liquid chromatography with tandem mass spectrometry was developed and optimized to quantify 21 amino acids and 9 water-soluble vitamins in Chinese hamster ovary (CHO) cells and culture media. By optimizing the chromatographic separation, scan time, monitoring time window, and sample preparation procedure, and using isotopically labeled (13)C, (15)N and (2)H internal standards, low limits of quantitation (≤0.054 mg/L), good precision (<10%) and good accuracy (100±10%) were achieved for nearly all the 30 compounds. Applying this method to CHO cell extracts, statistically significant differences in the metabolite levels were measured between two cell lines originated from the same host, indicating differences in genetic makeup or metabolic activities and nutrient supply levels in the culture media. In a fed-batch process of manufacturing scale bioreactors, two distinguished trends for changes in amino acid concentrations were identified in response to feeding. Ten essential amino acids showed a zigzag pattern with maxima at the feeding days, and 9 non-essential amino acids displayed a smoothly changing profile as they were mainly products of cellular metabolism. Five of 9 vitamins accumulated continuously during the culture period, suggesting that they were fed in access. The method serves as an effective tool for the development and optimization of mammalian cell cultures.

  4. The Utilization of Plant Facilities on the International Space Station-The Composition, Growth, and Development of Plant Cell Walls under Microgravity Conditions.

    PubMed

    Jost, Ann-Iren Kittang; Hoson, Takayuki; Iversen, Tor-Henning

    2015-01-01

    In the preparation for missions to Mars, basic knowledge of the mechanisms of growth and development of living plants under microgravity (micro-g) conditions is essential. Focus has centered on the g-effects on rigidity, including mechanisms of signal perception, transduction, and response in gravity resistance. These components of gravity resistance are linked to the evolution and acquisition of responses to various mechanical stresses. An overview is given both on the basic effect of hypergravity as well as of micro-g conditions in the cell wall changes. The review includes plant experiments in the US Space Shuttle and the effect of short space stays (8-14 days) on single cells (plant protoplasts). Regeneration of protoplasts is dependent on cortical microtubules to orient the nascent cellulose microfibrils in the cell wall. The space protoplast experiments demonstrated that the regeneration capacity of protoplasts was retarded. Two critical factors are the basis for longer space experiments: a. the effects of gravity on the molecular mechanisms for cell wall development, b. the availability of facilities and hardware for performing cell wall experiments in space and return of RNA/DNA back to the Earth. Linked to these aspects is a description of existing hardware functioning on the International Space Station. PMID:27135317

  5. A Global Information Utility.

    ERIC Educational Resources Information Center

    Block, Robert S.

    1984-01-01

    High-powered satellites, along with other existing technologies, make possible a world information utility that could distribute virtually limitless information to every point on earth. The utility could distribute information for business, government, education, and entertainment. How the utility would work is discussed. (RM)

  6. MIS Solar Cell Devices Based on a Cu2 O Substrate Utilizing h-BN as an Insulating and Passivating Layer

    NASA Astrophysics Data System (ADS)

    Ergen, Onur; Gibb, Ashley; Vazquez-Mena, Oscar; Regan, Will; Zettl, Alex

    2015-03-01

    We demonstrate Cu2O based metal insulator semiconductor Schottky (MIS-Schottky) solar cells with efficiency exceeding 3%. A unique direct growth technique is employed in the fabrication, and hexagonal boron nitride (h-BN) serves simultaneously as a passivation and insulation layer on the active cuprous oxide (Cu2O) layer. The devices are the most efficient of any Cu2O based MIS-Schottky solar cells reported to date. Department of Physics, University of California at Berkeley, Berkeley, California 94720, USA. Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA. Kavli Energy Nanosciences Institute at the University of Berkeley, CA, USA.

  7. Tissue-specific autoregulation of Drosophila suppressor of forked by alternative poly(A) site utilization leads to accumulation of the suppressor of forked protein in mitotically active cells.

    PubMed

    Juge, F; Audibert, A; Benoit, B; Simonelig, M

    2000-11-01

    The Suppressor of forked protein is the Drosophila homolog of the 77K subunit of human cleavage stimulation factor, a complex required for the first step of the mRNA 3'-end-processing reaction. We have shown previously that wild-type su(f) function is required for the accumulation of a truncated su(f) transcript polyadenylated in intron 4 of the gene. This led us to propose a model in which the Su(f) protein would negatively regulate its own accumulation by stimulating 3'-end formation of this truncated su(f) RNA. In this article, we demonstrate this model and show that su(f) autoregulation is tissue specific. The Su(f) protein accumulates at a high level in dividing tissues, but not in nondividing tissues. We show that this distribution of the Su(f) protein results from stimulation by Su(f) of the tissue-specific utilization of the su(f) intronic poly(A) site, leading to the accumulation of the truncated su(f) transcript in nondividing tissues. Utilization of this intronic poly(A) site is affected in a su(f) mutant and restored in the mutant with a transgene encoding wild-type Su(f) protein. These data provide an in vivo example of cell-type-specific regulation of a protein level by poly(A) site choice, and confirm the role of Su(f) in regulation of poly(A) site utilization.

  8. Utility of the Enzyme-Linked Immunospot Interferon-γ-Release Assay to Predict the Risk of Cytomegalovirus Infection in Hematopoietic Cell Transplant Recipients.

    PubMed

    Nesher, Lior; Shah, Dimpy P; Ariza-Heredia, Ella J; Azzi, Jacques M; Siddiqui, Hala K; Ghantoji, Shasank S; Marsh, Lisa Y; Michailidis, Lamprinos; Makedonas, George; Rezvani, Katy; Shpall, Elizabeth J; Chemaly, Roy F

    2016-06-01

    The ability to distinguish allogeneic hematopoietic cell transplant (allo-HCT) recipients at risk for cytomegalovirus (CMV) reactivation from those who are not is central for optimal CMV management strategies. Interferon γ (IFN-γ) produced by CMV-challenged T cells may serve as an immune marker differentiating these 2 populations. We prospectively monitored 63 CMV-seropositive allo-HCT recipients with a CMV-specific enzyme-linked immunospot (ELISPOT) assay and for CMV infection from the period before transplantation to day 100 after transplantation. Assay results above certain thresholds (50 spots per 250 000 cells for immediate early 1 or 100 spots per 250 000 cells for phosphoprotein 65) identified patients who were protected against CMV infection as long as they had no graft-versus-host disease and/or were not receiving systemic corticosteroids. Based on the multivariable Cox proportional hazards regression model, the only significant factor for preventing CMV reactivation was a CMV-specific ELISPOT response above the determined thresholds (adjusted hazard ratio, 0.21; 95% confidence interval, .05-.97; P = .046). Use of this assay as an additional tool for managing allo-HCT recipients at risk for CMV reactivation needs further validation in future studies. Application of this new approach may reduce the duration and intensity of CMV monitoring and the duration of prophylaxis or treatment with antiviral agents in those who have achieved CMV-specific immune reconstitution. PMID:26908740

  9. Investigating the utility of human embryonic stem cell-derived neurons to model ageing and neurodegenerative disease using whole-genome gene expression and splicing analysis

    PubMed Central

    Patani, Rickie; Lewis, Patrick A; Trabzuni, Daniah; Puddifoot, Clare A; Wyllie, David J A; Walker, Robert; Smith, Colin; Hardingham, Giles E; Weale, Michael; Hardy, John; Chandran, Siddharthan; Ryten, Mina

    2012-01-01

    A major goal in regenerative medicine is the predictable manipulation of human embryonic stem cells (hESCs) to defined cell fates that faithfully represent their somatic counterparts. Directed differentiation of hESCs into neuronal populations has galvanized much interest into their potential application in modelling neurodegenerative disease. However, neurodegenerative diseases are age-related, and therefore establishing the maturational comparability of hESC-derived neural derivatives is critical to generating accurate in vitro model systems. We address this issue by comparing genome-wide, exon-specific expression analyses of pluripotent hESCs, multipotent neural precursor cells and a terminally differentiated enriched neuronal population to expression data from post-mortem foetal and adult human brain samples. We show that hESC-derived neuronal cultures (using a midbrain differentiation protocol as a prototypic example of lineage restriction), while successful in generating physiologically functional neurons, are closer to foetal than adult human brain in terms of molecular maturation. These findings suggest that developmental stage has a more dominant influence on the cellular transcriptome than regional identity. In addition, we demonstrate that developmentally regulated gene splicing is common, and potentially a more sensitive measure of maturational state than gene expression profiling alone. In summary, this study highlights the value of genomic indices in refining and validating optimal cell populations appropriate for modelling ageing and neurodegeneration. PMID:22681703

  10. Bare and protein-conjugated Fe3O4 ferromagnetic nanoparticles for utilization in magnetically assisted hemodialysis: biocompatibility with human blood cells

    NASA Astrophysics Data System (ADS)

    Stamopoulos, D.; Manios, E.; Gogola, V.; Benaki, D.; Bouziotis, P.; Niarchos, D.; Pissas, M.

    2008-12-01

    Magnetically assisted hemodialysis is a development of conventional hemodialysis and is based on the circulation of ferromagnetic nanoparticle-targeted binding substance conjugates (FN-TBS Cs) in the bloodstream of the patient and their eventual removal by means of a 'magnetic dialyzer'. Presented here is an in vitro investigation on the biocompatibility of bare Fe3O4 FNs and Fe3O4-bovine serum albumin Cs with blood cells, namely red blood cells (RBCs), white blood cells (WBCs) and platelets (Plts). Atomic force microscopy (AFM) and optical microscopy (OM) enabled the examination of blood cells at the nanometer and micrometer level, respectively. The observations made on FN- and C-maturated blood samples are contrasted to those obtained on FN- and C-free reference blood samples subjected to exactly the same maturation procedure. Qualitatively, both AFM and OM revealed no changes in the overall shape of RBCs, WBCs and Plts. Incidents where bare FNs or Cs were bound onto the surface of RBCs or internalized by WBCs were very rare. Detailed examination by means of OM proved that impaired coagulation of Plts is not initiated/promoted either by FNs or Cs. Quantitatively, the statistical analysis of the obtained AFM images from RBC surfaces clearly revealed that the mean surface roughness of RBCs maturated with bare FNs or Cs was identical to the one of reference RBCs.

  11. Novel Antitumor Strategy Utilizing a Plasmid Expressing a Mycobacterium tuberculosis Antigen as a “Danger Signal” to Block Immune Escape of Tumor Cells

    PubMed Central

    Koyama, Yoshiyuki; Yoshihara, Chieko; Ito, Tomoko

    2015-01-01

    Immune escape of tumor cells is one of the main obstacles hindering the effectiveness of cancer immunotherapy. We developed a novel strategy to block immune escape by transfecting tumor cells in vivo with genes of pathogenic antigens from Mycobacterium tuberculosis (TB). This induces presentation of the TB antigen on tumor cell surfaces, which can be recognized by antigen presenting cells (APCs) as a “danger signal” to stimulate antitumor immune response. This strategy is also expected to amplify the immune response against tumor-associated antigens, and block immune escape of the tumor. DNA/PEI/chondroitin sulfate ternary complex is a highly effective non-viral gene vector system for in vivo transfection. A therapeutic complex was prepared using a plasmid encoding the TB antigen, early secretory antigenic target-6 (ESAT-6). This was injected intratumorally into syngeneic tumor-bearing mice, and induced significant tumor growth suppression comparable to or higher than similar complexes expressing cytokines such as interleukin-2 (IL-2) and interleukin-12 (IL-12). Co-transfection of the cytokine-genes and the ESAT-6-gene enhanced the antitumor efficacy of either treatment alone. In addition, complete tumor regression was achieved with the combination of ESAT-6 and IL-2 genes. PMID:26213962

  12. Novel Antitumor Strategy Utilizing a Plasmid Expressing a Mycobacterium tuberculosis Antigen as a "Danger Signal" to Block Immune Escape of Tumor Cells.

    PubMed

    Koyama, Yoshiyuki; Yoshihara, Chieko; Ito, Tomoko

    2015-07-24

    Immune escape of tumor cells is one of the main obstacles hindering the effectiveness of cancer immunotherapy. We developed a novel strategy to block immune escape by transfecting tumor cells in vivo with genes of pathogenic antigens from Mycobacterium tuberculosis (TB). This induces presentation of the TB antigen on tumor cell surfaces, which can be recognized by antigen presenting cells (APCs) as a "danger signal" to stimulate antitumor immune response. This strategy is also expected to amplify the immune response against tumor-associated antigens, and block immune escape of the tumor. DNA/PEI/chondroitin sulfate ternary complex is a highly effective non-viral gene vector system for in vivo transfection. A therapeutic complex was prepared using a plasmid encoding the TB antigen, early secretory antigenic target-6 (ESAT-6). This was injected intratumorally into syngeneic tumor-bearing mice, and induced significant tumor growth suppression comparable to or higher than similar complexes expressing cytokines such as interleukin-2 (IL-2) and interleukin-12 (IL-12). Co-transfection of the cytokine-genes and the ESAT-6-gene enhanced the antitumor efficacy of either treatment alone. In addition, complete tumor regression was achieved with the combination of ESAT-6 and IL-2 genes.

  13. Urinary bladder smooth muscle regeneration utilizing bone marrow derived mesenchymal stem cell seeded elastomeric poly(1,8-octanediol-co-citrate) based thin films.

    PubMed

    Sharma, Arun K; Hota, Partha V; Matoka, Derek J; Fuller, Natalie J; Jandali, Danny; Thaker, Hatim; Ameer, Guillermo A; Cheng, Earl Y

    2010-08-01

    Bladder regeneration studies have yielded inconclusive results possibly due to the use of unfavorable cells and primitive scaffold design. We hypothesized that human mesenchymal stem cells seeded onto poly(1,8-octanediol-co-citrate) elastomeric thin films would provide a suitable milieu for partial bladder regeneration. POCfs were created by reacting citric acid with 1,8-octanediol and seeded on opposing faces with human MSCs and urothelial cells; normal bladder smooth muscle cells and UCs, or unseeded POCfs. Partial cystectomized nude rats were augmented with the aforementioned POCfs, enveloped with omentum and sacrificed at 4 and 10 weeks. Isolated bladders were subjected to Trichrome and anti-human gamma-tubulin, calponin, caldesmon, smooth muscle gamma-actin, and elastin stainings. Mechanical testing of POCfs revealed a Young's modulus of 138 kPa with elongation 137% its initial length without permanent deformation demonstrating its high uniaxial elastic potential. Trichrome and immunofluorescent staining of MSC/UC POCf augmented bladders exhibited typical bladder architecture with muscle bundle formation and the expression and retention of bladder smooth muscle contractile proteins of human derivation. Quantitative morphometry of MSC/UC samples revealed muscle/collagen ratios approximately 1.75x greater than SMC/UC controls at 10 weeks. Data demonstrate MSC seeded POCfs support partial regeneration of bladder tissue in vivo.

  14. Utility of the Enzyme-Linked Immunospot Interferon-γ–Release Assay to Predict the Risk of Cytomegalovirus Infection in Hematopoietic Cell Transplant Recipients

    PubMed Central

    Nesher, Lior; Shah, Dimpy P.; Ariza-Heredia, Ella J.; Azzi, Jacques M.; Siddiqui, Hala K.; Ghantoji, Shasank S.; Marsh, Lisa Y.; Michailidis, Lamprinos; Makedonas, George; Rezvani, Katy; Shpall, Elizabeth J.; Chemaly, Roy F.

    2016-01-01

    The ability to distinguish allogeneic hematopoietic cell transplant (allo-HCT) recipients at risk for cytomegalovirus (CMV) reactivation from those who are not is central for optimal CMV management strategies. Interferon γ (IFN-γ) produced by CMV-challenged T cells may serve as an immune marker differentiating these 2 populations. We prospectively monitored 63 CMV-seropositive allo-HCT recipients with a CMV-specific enzyme-linked immunospot (ELISPOT) assay and for CMV infection from the period before transplantation to day 100 after transplantation. Assay results above certain thresholds (50 spots per 250 000 cells for immediate early 1 or 100 spots per 250 000 cells for phosphoprotein 65) identified patients who were protected against CMV infection as long as they had no graft-versus-host disease and/or were not receiving systemic corticosteroids. Based on the multivariable Cox proportional hazards regression model, the only significant factor for preventing CMV reactivation was a CMV-specific ELISPOT response above the determined thresholds (adjusted hazard ratio, 0.21; 95% confidence interval, .05–.97; P = .046). Use of this assay as an additional tool for managing allo-HCT recipients at risk for CMV reactivation needs further validation in future studies. Application of this new approach may reduce the duration and intensity of CMV monitoring and the duration of prophylaxis or treatment with antiviral agents in those who have achieved CMV-specific immune reconstitution. PMID:26908740

  15. Differential effects of "Advanced glycation endproducts" and beta-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y.

    PubMed

    Kuhla, B; Loske, C; Garcia De Arriba, S; Schinzel, R; Huber, J; Münch, G

    2004-03-01

    Beta-amyloid peptide (Abeta) and "Advanced glycation endproducts" (AGEs) are components of the senile plaques in Alzheimer's disease patients. It has been proposed that both AGEs and Abeta exert many of their effects, which include the upregulation of pro-inflammatory cytokines, through RAGE ("receptor for advanced glycation endproducts"). To investigate whether Abeta and AGEs cause similar or identical effects on cell survival and energy metabolism, we have compared the effects of a model-AGE and Abeta on cell viability, ATP level, glucose consumption and lactate production in the neuroblastoma cell line SH-SY5Y. The results show that AGEs and Abeta increase glucose consumption and decrease ATP levels in a dose dependent manner. Furthermore, both compounds decrease mitochondrial activity measured by the MTT assay. However, only AGEs decrease the number of cells and significantly increase lactate production. These data indicate that both AGEs and Abeta can cause differential disturbances in neuronal metabolism, which may contribute to the pathophysiological findings in Alzheimer's disease. However, their signalling pathways are apparently quite distinct, a fact which should stimulate a more detailed investigation in this field, e.g. for the purpose of a rational design of potential "neuroprotective" RAGE antagonists. PMID:14991463

  16. A robust one-compartment fuel cell with a polynuclear cyanide complex as a cathode for utilizing H2O2 as a sustainable fuel at ambient conditions.

    PubMed

    Yamada, Yusuke; Yoneda, Masaki; Fukuzumi, Shunichi

    2013-08-26

    A robust one-compartment H2O2 fuel cell, which operates without membranes at room temperature, has been constructed by using a series of polynuclear cyanide complexes that contain Fe, Co, Mn, and Cr as cathodes, in sharp contrast to conventional H2 and MeOH fuel cells, which require membranes and high temperatures. A high open-circuit potential of 0.68 V was achieved by using Fe3[{Co(III)(CN)6}2] on a carbon cloth as the cathode and a Ni mesh as the anode of a H2O2 fuel cell by using an aqueous solution of H2O2 (0.30  M, pH 3) with a maximum power density of 0.45 mW cm(-2). The open-circuit potential and maximum power density of the H2O2 fuel cell were further increased to 0.78 V and 1.2 mW cm(-2), respectively, by operation under these conditions at pH 1. No catalytic activity of Co3[{Co(III)(CN)6}2] and Co3[{Fe(III)(CN)6}2] towards H2O2 reduction suggests that the N-bound Fe ions are active species for H2O2 reduction. H2O2 fuel cells that used Fe3[{Mn(III)(CN)6}2] and Fe3[{Cr(III)(CN)6}2] as the cathode exhibited lower performance compared with that using Fe3[{Co(III)(CN)6}2] as a cathode, because ligand isomerization of Fe3[{M(III)(CN)6}2] into (FeM2)[{Fe(II)(CN)6}2] (M = Cr or Mn) occurred to form inactive Fe-C bonds under ambient conditions, whereas no ligand isomerization of Fe3[{Co(III)(CN)6}2] occurred under the same reaction conditions. The importance of stable Fe(2+)-N bonds was further indicated by the high performance of the H2O2 fuel cells with Fe3[{Ir(III)(CN)6}2] and Fe3[{Rh(III)(CN)6}2], which also contained stable Fe(2+)-N bonds. The stable Fe(2+)-N bonds in Fe3[{Co(III)(CN)6}2], which lead to high activity for the electrocatalytic reduction of H2O2, allow Fe3[{Co(III)(CN)6}2] to act as a superior cathode in one-compartment H2O2 fuel cells.

  17. A robust one-compartment fuel cell with a polynuclear cyanide complex as a cathode for utilizing H2O2 as a sustainable fuel at ambient conditions.

    PubMed

    Yamada, Yusuke; Yoneda, Masaki; Fukuzumi, Shunichi

    2013-08-26

    A robust one-compartment H2O2 fuel cell, which operates without membranes at room temperature, has been constructed by using a series of polynuclear cyanide complexes that contain Fe, Co, Mn, and Cr as cathodes, in sharp contrast to conventional H2 and MeOH fuel cells, which require membranes and high temperatures. A high open-circuit potential of 0.68 V was achieved by using Fe3[{Co(III)(CN)6}2] on a carbon cloth as the cathode and a Ni mesh as the anode of a H2O2 fuel cell by using an aqueous solution of H2O2 (0.30  M, pH 3) with a maximum power density of 0.45 mW cm(-2). The open-circuit potential and maximum power density of the H2O2 fuel cell were further increased to 0.78 V and 1.2 mW cm(-2), respectively, by operation under these conditions at pH 1. No catalytic activity of Co3[{Co(III)(CN)6}2] and Co3[{Fe(III)(CN)6}2] towards H2O2 reduction suggests that the N-bound Fe ions are active species for H2O2 reduction. H2O2 fuel cells that used Fe3[{Mn(III)(CN)6}2] and Fe3[{Cr(III)(CN)6}2] as the cathode exhibited lower performance compared with that using Fe3[{Co(III)(CN)6}2] as a cathode, because ligand isomerization of Fe3[{M(III)(CN)6}2] into (FeM2)[{Fe(II)(CN)6}2] (M = Cr or Mn) occurred to form inactive Fe-C bonds under ambient conditions, whereas no ligand isomerization of Fe3[{Co(III)(CN)6}2] occurred under the same reaction conditions. The importance of stable Fe(2+)-N bonds was further indicated by the high performance of the H2O2 fuel cells with Fe3[{Ir(III)(CN)6}2] and Fe3[{Rh(III)(CN)6}2], which also contained stable Fe(2+)-N bonds. The stable Fe(2+)-N bonds in Fe3[{Co(III)(CN)6}2], which lead to high activity for the electrocatalytic reduction of H2O2, allow Fe3[{Co(III)(CN)6}2] to act as a superior cathode in one-compartment H2O2 fuel cells. PMID:23868499

  18. Utilization of proteinaceous materials for power generation in a mediatorless microbial fuel cell by a new electrogenic bacteria Lysinibacillus sphaericus VA5.

    PubMed

    Nandy, Arpita; Kumar, Vikash; Kundu, Patit P

    2013-10-10

    In this study, a bacterial strain, Lysinibacillus sphaericus which is relatively new in the vast list of biocatalysts known to produce electricity has been tested for its potential in power production. It is cited from the literature that the organism is deficient in some sugar or polysaccharide processing enzymes and thus is tested for its ability to utilize substrates mainly rich in protein components like beef extract and with successive production of electricity. The particular species has been found to generate a maximum power density of 85mW/m(2) and current density of ≈270mA/m(2) using graphite felt as electrode. The maximum Open Circuit Voltage and current has been noted as 0.7Vand 0.8mA during these operational cycles. Cyclic voltammetry studies indicate the presence of some electroactive compounds which can facilitate electron transfer from bacteria to electrode. The number of electrogens able to generate electricity in mediator free conditions are few, and the study introduces more divergence to that population. Substrate specificity and electricity generation efficacy of the strain in treating wastewater, specially rich in protein content has been reported in the study. As the species has been found to be efficient in utilizing proteinaceous material, the technique can be useful to treat specific type of wastewaters like wastewater from slaughterhouses or from meat packaging industry. Treating them in a more economical way which generates electricity as a outcome must be preferred over the conventional aerobic treatments. Emphasizing on substrate specificity, the study introduces this novel Lysinibacillus strain as a potent biocatalyst and its sustainable role in MFC application for bioenergy generation. PMID:24034433

  19. Transforming growth factor-beta2 utilizes the canonical Smad-signaling pathway to regulate tissue transglutaminase expression in human trabecular meshwork cells.

    PubMed

    Tovar-Vidales, Tara; Clark, Abbot F; Wordinger, Robert J

    2011-10-01

    Transforming growth factor-beta2 (TGF-β2) is elevated in the aqueous humor of patients with glaucoma. This growth factor is known to increase extracellular matrix (ECM) deposition in the trabecular meshwork (TM) as well as increase intraocular pressure (IOP) in perfused human cultured anterior eye segments. In addition overexpression of TGF-β2 in the mouse TM leads to elevated IOP. Exogenous TGF-β2 also increases tissue transglutaminase (TGM2) protein levels and enzyme activity in TM cells. TGM2 is a calcium-dependent enzyme that mediates cross-linking of ECM proteins, thus making ECM proteins resistant to enzymatic degradation and physical breakdown. We have investigated the signaling pathway by which TGF-β2 induces TGM2 in human TM cells. Primary cultures of human TM cells (N = 6) were treated for 48 h with TGF-β2 (0-10 ng/ml) in serum-free medium. TGM2 enzyme activity differences between non-treated and TGF-β2 treated TM cells were studied using a biotin cadaverine assay. Endogenous TGF-β2 protein levels were examined in normal trabecular meshwork (NTM) and glaucomatous trabecular meshwork (GTM) cell strains. Immunohistochemistry was used to evaluate the expression and co-localization of TGF-β2 and TGM2 in NTM and GTM tissues. Activation of Smad3 signaling pathway was evaluated by western immunoblot analysis using phospho-specific antibodies following exogenous TGF-β2 treatment. Pharmacological specific inhibitor of Smad3 (SIS3) and short interfering (si)RNAs were used to suppress Smad3 activity and CTGF gene expression respectively. Endogenous TGF-β2 levels were significantly elevated in cultured GTM cells (p < 0.05) when compared to NTM cells. Immunohistochemistry studies also demonstrated elevated expression and co-localization of both TGF-β2 and TGM2 in glaucoma human TM tissues. Exogenous TGF-β2 increased both TGM2 protein levels and enzyme activity in TM cells. Phosphorylation of Smad3 was stimulated in TM cell strains by exogenous TGF

  20. Low-cost, high-efficiency solar cells utilizing GaAs-on-Si technology. Annual subcontract report, 1 August 1991--31 July 1992

    SciTech Connect

    Vernon, S.M.

    1993-04-01

    This report describes work to develop technology to deposit GaAs on Si using a nucleation layer of atomic-layer-epitaxy-grown GaAs or AlAs on Si. This ensures two-dimensional nucleation and should lead to fewer defects in the final GaAs layer. As an alternative, we also developed technology for depositing GaAs on sawtooth-patterned Si. Preliminary studies showed that this material can have a very low defect density, {approximately} 1 {times} 10{sup 5} cm{sup {minus}5}, as opposed to our conventionally grown GaAs on SL which has a typical defect density of over 1 {times}10{sup 7} cm{sup {minus}2}. Using these two now methods of GaAs-on-Si material growth, we made solar cells that are expected to show higher efficiencies than those of previous cells.

  1. Fabrication of cell penetration enhanced poly (l-lactic acid-co-ɛ-caprolactone)/silk vascular scaffolds utilizing air-impedance electrospinning.

    PubMed

    Yin, Anlin; Li, Jiukai; Bowlin, Gary L; Li, Dawei; Rodriguez, Isaac A; Wang, Jing; Wu, Tong; Ei-Hamshary, Hany A; Al-Deyab, Salem S; Mo, Xiumei

    2014-08-01

    In the vascular prosthetic field, the prevailing thought is that for clinical, long-term success, especially bioresorbable grafts, cellular migration and penetration into the prosthetic structure is required to promote neointima formation and vascular wall development. In this study, we fabricated poly (l-lactic acid-co-ɛ-caprolactone) P(LLA-CL)/silk fibroin (SF) vascular scaffolds through electrospinning using both perforated mandrel subjected to various intraluminal air pressures (0-300kPa), and solid mandrel. The scaffolds were evaluated the cellular infiltration in vitro and mechanical properties. Vascular scaffolds were seeded with smooth muscle cells (SMCs) to evaluate cellular infiltration at 1, 7, and 14 days. The results revealed that air-impedance scaffolds allowed significantly more cell infiltration as compared to the scaffolds fabricated with solid mandrel. Meanwhile, results showed that both mandrel model and applied air pressure determined the interfiber distance and the alignment of fibers in the enhanced porosity regions of the structure which influenced cell infiltration. Uniaxial tensile testing indicated that the air-impedance scaffolds have sufficient ultimate strength, suture retention strength, and burst pressure as well as compliance approximating a native artery. In conclusion, the air-impedance scaffolds improved cellular infiltration without compromising overall biomechanical properties. These results support the scaffold's potential for vascular grafting and in situ regeneration.

  2. Phosphorylation of the alternative mRNA splicing factor 45 (SPF45) by Clk1 regulates its splice site utilization, cell migration and invasion

    PubMed Central

    Liu, Yuying; Conaway, LaShardai; Rutherford Bethard, Jennifer; Al-Ayoubi, Adnan M.; Thompson Bradley, Amber; Zheng, Hui; Weed, Scott A.; Eblen, Scott T.

    2013-01-01

    Alternative mRNA splicing is a mechanism to regulate protein isoform expression and is regulated by alternative splicing factors. The alternative splicing factor 45 (SPF45) is overexpressed in cancer, although few biological effects of SPF45 are known, and few splicing targets have been identified. We previously showed that Extracellular Regulated Kinase 2 (ERK2) phosphorylation of SPF45 regulates cell proliferation and adhesion to fibronectin. In this work, we show that Cdc2-like kinase 1 (Clk1) phosphorylates SPF45 on eight serine residues. Clk1 expression enhanced, whereas Clk1 inhibition reduced, SPF45-induced exon 6 exclusion from Fas mRNA. Mutational analysis of the Clk1 phosphorylation sites on SPF45 showed both positive and negative regulation of splicing, with a net effect of inhibiting SPF45-induced exon 6 exclusion, correlating with reduced Fas mRNA binding. However, Clk1 enhanced SPF45 protein expression, but not mRNA expression, whereas inhibition of Clk1 increased SPF45 degradation through a proteasome-dependent pathway. Overexpression of SPF45 or a phospho-mimetic mutant, but not a phospho-inhibitory mutant, stimulated ovarian cancer cell migration and invasion, correlating with increased fibronectin expression, ERK activation and enhanced splicing and phosphorylation of full-length cortactin. Our results demonstrate for the first time that SPF45 overexpression enhances cell migration and invasion, dependent on biochemical regulation by Clk1. PMID:23519612

  3. Utilization of monoclonal-antibody-based assay (HemoCard in screening for and differentiating between genotypes of sickle cell disease and other hemoglobinopathies.

    PubMed

    Schultz, J C

    1995-01-01

    Sickle cell disease covers a group of conditions in which pathology may be attributed to the presence of sickle hemoglobin (HbS). The identification of HbS and other variants including those in combination with HbS is commonly achieved by cellulose acetate electrophoresis at alkaline pH. Because many hemoglobin variants with similar charges have similar electrophoretic migration patterns, they are difficult to differentiate by electrophoresis. The HemoCard assays address this concern through the use of monoclonal antibodies capable of specifically recognizing the unique amino acid substitution in the variant hemoglobin. The panel of HemoCard monoclonal antibodies confirms the absence and presence of HbA, HbC, HbE, HbS, and other sickling hemoglobin variants. The combination of alkaline cellulose acetate electrophoresis and HemoCard assays allows the technologist to reach a final conformation of both common and much less common sickle cell disease genotypes, combinations of HbS with other hemoglobins that ordinarily do not produce sickle cell disease, and other clinically important hemoglobinopathies including HbE/beta-thalassemia and hemoglobin C disease. PMID:8587004

  4. Utilizing muscle-derived stem cells to enhance long-term retention and aesthetic outcome of autologous fat grafting: pilot study in mice.

    PubMed

    Ma, Zhiqiang; Han, Duanyang; Zhang, Peipei; Yang, Jenny F; Wang, Yiqiang; Zhang, Yingbo; Yang, Daping; Liu, Jianyu

    2012-02-01

    Autologous fat grafting has been regarded as the ideal soft tissue filler for more than a century. Low long-term retention rate and unpredictability limit it from widespread clinical practice. Many theories for this have been proposed: lack of sufficient blood supply and subsequent necrosis is the most accepted. In this pilot study, we showed both macroscopically and microscopically the viability of muscle-derived stem cells (MDSCs) cotransplanted with fat placed intramuscularly for 3 months. MRI scanning showed a stronger fat signal in the MDSC-treated group than that of the control group. Moreover, histological evaluation exhibited well-preserved and intact fat cells in the MDSC-treated group. In contrast, the control group showed extensive fibrosis and fat graft loss. Furthermore, the MDSC-treated group possessed almost threefold greater capillary density than the control group. We conclude that cotransplantation of muscle-derived stem cells and autologous fat tissue improves the long-term survival of intramuscular fat transplants by promoting neovascularization. PMID:21607534

  5. High-performance inverted tandem polymer solar cells utilizing thieno[3,4-c]pyrrole-4,6-dione copolymer.

    PubMed

    Yusoff, Abd Rashid bin Mohd; Lee, Seung Joo; Kim, Jaeyeon; Shneider, Fabio Kurt; da Silva, Wilson Jose; Jang, Jin

    2014-08-13

    We demonstrated the inverted solution processed tandem polymer solar cells, in which transparent pH-neutral poly(3,4-ethylenedioxylenethiophene)-polystylene sulfonic acid (PEDOT:PSS) and lithium zinc oxide layers were used as a recombination layer. We have used poly(di(2-ethylhexyloxy)benzo[1,2-b:4,5-b']dithiophene-co-octylthieno[3,4-c]pyrrole-4,6-dione):[6,6]-phenyl-C61 butyric acid methyl ester (PBDTTPD:PC61BM) and poly[(4,4'-bis(2-ethylhexyl)dithieno[3,2-b:2',3'-d] silole)-2,6-diyl-alt-(2,1,3-benzothiadiazole)-4,7-diyl]:[6,6]-phenyl-C70 butyric acid methyl ester (PSBTBT:PC70BM) as the active layers for front and rear subcells, respectively. The pH-neutral PEDOT:PSS/LZO serves as an electron- and hole-collecting and recombination layer. Our tandem solar cells showed a high open circuit voltage (Voc) of 1.54 V, a short circuit current density (Jsc) of 7.55 mA/cm(2), and a fill factor (FF) of 64.79% along with the power conversion efficiency of 7.53%. The Voc value of our tandem solar cells is an ideal summation of Voc values from front and rear subcells. PMID:24967661

  6. Stability improvement of gel-state dye-sensitized solar cells by utilization the co-solvent effect of propionitrile/acetonitrile and 3-methoxypropionitrile/acetonitrile with poly(acrylonitrile-co-vinyl acetate)

    NASA Astrophysics Data System (ADS)

    Venkatesan, Shanmugam; Su, Song-Chuan; Kao, Shon-Chen; Teng, Hsisheng; Lee, Yuh-Lang

    2015-01-01

    Propionitrile (PPN) or 3-methoxypropionitrile (MPN) is mixed with acetonitrile (ACN) to prepare ACN/PPN and ACN/MPN co-solvents and used to fabricate polymer gel electrolytes (PGEs) of dye-sensitized solar cells (DSSCs), aiming at improving the stability of gel-state DSSCs. Co-solvents with various ratios are utilized to prepare PGEs using poly(acrylonitrile-co-vinyl acetate) (PAN-VA) as the gelator. The ratio effects of the co-solvents on the properties of PGEs and the performances of the corresponding DSSCs are studied. The results show that in-situ gelation of the gel-electrolytes can still be performed at the presence of 40% PPN or 30% MPN. However, increasing the composition of PPN and MPN in the co-solvents triggers a decrease in the diffusivity and conductivity of the PGEs, but an increase in the viscosity. Therefore, the energy conversion efficiencies of the cells decrease as a result. However, the introduction of PPN and MPN elevates the gel-to-liquid transition temperature (Tp) of the PGEs which significantly increases the stability of the gel-state DSSCs. Comparing between the effects of the two co-solvents, PPN and MPN have similar effect on elevation of Tp, but the conductivity of PGEs and the corresponding cell efficiency are higher for the ACN/PPN system, attributed to its lower viscosity compared with ACN/MPN system. By using the ACN/PPN (60/40) co-solvent at the presence of TiO2 fillers, gel-state cell with an efficiency of 8.3% can be achieved, which is even higher than that obtained by the liquid state cell (8%). After 500 h test at 60 °C, the cell can retain 95.4% of its initial efficiency.

  7. Distributed H2 Supply for Fuel Cell Utility Vehicles Year 6 - Activity 3.5 - Development fo a National Center for Hydrogen Technology

    SciTech Connect

    Almlie, Jay

    2012-04-15

    The Energy & Environmental Research Center (EERC) has developed a high-pressure hydrogen production system that reforms a liquid organic feedstock and water at operating pressures up to 800 bar (~12,000 psig). The advantages of this system include the elimination of energy-intensive hydrogen compression, a smaller process footprint, and the elimination of gaseous or liquid hydrogen transport. This system could also potentially enable distributed hydrogen production from centralized coal. Processes have been investigated to gasify coal and then convert the syngas into alcohol or alkanes. These alcohols and alkanes could then be easily transported in bulk to distributed high-pressure water-reforming (HPWR)-based systems to deliver hydrogen economically. The intent of this activity was to utilize the EERC’s existing HPWR hydrogen production process, previously designed and constructed in a prior project phase, as a basis to improve operational and production performance of an existing demonstration unit. Parameters to be pursued included higher hydrogen delivery pressure, higher hydrogen production rates, and the ability to refill within a 5-minute time frame.

  8. Thermodynamics of sulfur poisoning in solid oxide fuel cells revisited: The effect of H2S concentration, temperature, current density and fuel utilization

    NASA Astrophysics Data System (ADS)

    da Silva, Aline Lima; Heck, Nestor Cezar

    2015-11-01

    Thermodynamics of sulfur poisoning in SOFCs is revisited in the present study, aiming at contributing to the understanding of the effects of different operating parameters on deactivation by sulfur. Ni-S Gibbs energy diagram shows that, whenever sulfur chemical potential is increased, the catalyst poisoning becomes greater, due to increase in nickel sulfide activity, which is a strictly increasing function of sulfur coverage on Ni surface. For studying current density effect, simulations are carried out in the range of 0 (OCV) - 1 A cm-2, at 1123 K, considering methane as a fuel. At 10 ppm H2S, an increase in current density from 0 (OCV) to 0.5 A cm-2 results in a slight increase in the nickel sulfide activity from 3.0 × 10-6 to 1.2 × 10-5; however, at 1 A cm-2, nickel sulfide activity is 25.000 times higher than at 0.5 A cm-2. At 1 A cm-2, H2 and CO are almost entirely converted to H2O and CO2 by electrochemical reaction. Therefore, the effect of current density becomes remarkable when fuel utilization approaches 100%. These theoretical findings are corroborated by the recent experimental evidences related to the detrimental influence of current density on SOFC performance during sulfur poisoning.

  9. Monoclonal antibodies, carbohydrate-binding modules, and the detection of polysaccharides in plant cell walls.

    PubMed

    Hervé, Cécile; Marcus, Susan E; Knox, J Paul

    2011-01-01

    Plant cell walls are diverse composites of complex polysaccharides. Molecular probes such as monoclonal antibodies (MABs) and carbohydrate-binding modules (CBMs) are important tools to detect and dissect cell wall structures in plant materials. We provide an account of methods that can be used to detect cell wall polysaccharide structures (epitopes) in plant materials and also describe treatments that can provide information on the masking of sets of polysaccharides that may prevent detection. These masking -phenomena may indicate potential interactions between sets of cell wall polysaccharides, and methods to uncover them are an important aspect of cell wall immunocytochemistry.

  10. Construction of engineering adipose-like tissue in vivo utilizing human insulin gene-modified umbilical cord mesenchymal stromal cells with silk fibroin 3D scaffolds.

    PubMed

    Li, Shi-Long; Liu, Yi; Hui, Ling

    2015-12-01

    We evaluated the use of a combination of human insulin gene-modified umbilical cord mesenchymal stromal cells (hUMSCs) with silk fibroin 3D scaffolds for adipose tissue engineering. In this study hUMSCs were isolated and cultured. HUMSCs infected with Ade-insulin-EGFP were seeded in fibroin 3D scaffolds with uniform 50-60 µm pore size. Silk fibroin scaffolds with untransfected hUMSCs were used as control. They were cultured for 4 days in adipogenic medium and transplanted under the dorsal skins of female Wistar rats after the hUMSCs had been labelled with chloromethylbenzamido-1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (CM-Dil). Macroscopical impression, fluorescence observation, histology and SEM were used for assessment after transplantation at 8 and 12 weeks. Macroscopically, newly formed adipose tissue was observed in the experimental group and control group after 8 and 12 weeks. Fluorescence observation supported that the formed adipose tissue originated from seeded hUMSCs rather than from possible infiltrating perivascular tissue. Oil red O staining of newly formed tissue showed that there was substantially more tissue regeneration in the experimental group than in the control group. SEM showed that experimental group cells had more fat-like cells, whose volume was larger than that of the control group, and degradation of the silk fibroin scaffold was greater under SEM observation. This study provides significant evidence that hUMSCs transfected by adenovirus vector have good compatibility with silk fibroin scaffold, and adenoviral transfection of the human insulin gene can be used for the construction of tissue-engineered adipose.

  11. Utilities weather the storm

    SciTech Connect

    Lihach, N.

    1984-11-01

    Utilities must restore power to storm-damaged transmission and distribution systems, even if it means going out in ice storms or during lightning and hurricane conditions. Weather forecasting helps utilities plan for possible damage as well as alerting them to long-term trends. Storm planning includes having trained repair personnel available and adjusting the system so that less power imports are needed. Storm damage response requires teamwork and cooperation between utilities. Utilities can strengthen equipment in storm-prone or vulnerable areas, but good data are necessary to document the incidence of lighning strikes, hurricanes, etc. 2 references, 8 figures.

  12. Utilization of an automated pipetting system in the cell line-based screening of the activity of a DNA-damaging anti-tumour drug.

    PubMed

    Suchánková, T; Ovesná, P; Samadder, P; Souček, K

    2014-01-01

    The principles of large screening strategies, which are developed by industrial companies, have been recently adopted by researchers in the fields of molecular biology and oncology as invaluable tools for translational medicine. The declining costs of laboratory robotic machines have allowed high-throughput screening to become more available to academic centres with limited resources. Here, we describe how a robotic conventional liquid handling system could be used on a daily basis in laboratories to obtain consistent and reproducible results. Our approach allowed us to quickly screen a panel of more than 20 tumorigenic and non-tumorigenic cell lines for their responses to hydroxyurea, which is a DNA-damaging anticancer therapeutic drug. The format of 384-well microplates was used for manual cell seeding, and the effect of hydroxyurea was screened at multiple concentrations. The fluorescence-based CyQuant assay was employed as the readout method to analyse the cellular DNA content. The effectiveness of our approach was demonstrated in the experimental results.

  13. Utilization of Amygdalin during Seedling Development of Prunus serotina.

    PubMed

    Swain, E.; Poulton, J. E.

    1994-10-01

    Cotyledons of mature black cherry (Prunus serotina Ehrh.) seeds contain the cyanogenic diglucoside (R)-amygdalin. The levels of amygdalin, its corresponding monoglucoside (R)-prunasin, and the enzymes that metabolize these cyanoglycosides were measured during the course of seedling development. During the first 3 weeks following imbibition, cotyledonary amygdalin levels declined by more than 80%, but free hydrogen cyanide was not released to the atmosphere. Concomitantly, prunasin, which was not present in mature, ungerminated seeds, accumulated in the seedling epicotyls, hypocotyls, and cotyledons to levels approaching 4 [mu]mol per seedling. Whether this prunasin resulted from amygdalin hydrolysis remains unclear, however, because these organs also possess UDPG:mandelonitrile glucosyltransferase, which catalyzes de novo prunasin biosynthesis. The reduction in amygdalin levels was paralleled by declines in the levels of amygdalin hydrolase (AH), prunasin hydrolase (PH), mandelonitrile lyase (MDL), and [beta]-cyanoalanine synthase. At all stages of seedling development, AH and PH were localized by immunocytochemistry within the vascular tissues. In contrast, MDL occurred mostly in the cotyledonary parenchyma cells but was also present in the vascular tissues. Soon after imbibition, AH, PH, and MDL were found within protein bodies but were later detected in vacuoles derived from these organelles. PMID:12232341

  14. I-III-VI.sub.2 based solar cell utilizing the structure CuInGaSe.sub.2 CdZnS/ZnO

    DOEpatents

    Chen, Wen S.; Stewart, John M.

    1992-01-07

    A thin film I-III-VI.sub.2 based solar cell having a first layer of copper indium gallium selenide, a second layer of cadmium zinc sulfide, a double layer of zinc oxide, and a metallization structure comprised of a layer of nickel covered by a layer of aluminum. An optional antireflective coating may be placed on said metallization structure. The cadmium zinc sulfide layer is deposited by means of an aqueous solution growth deposition process and may actually consist of two layers: a low zinc content layer and a high zinc content layer. Photovoltaic efficiencies of 12.5% at Air Mass 1.5 illumination conditions and 10.4% under AMO illumination can be achieved.

  15. Utility of GATA3 immunohistochemistry in differentiating urothelial carcinoma from prostate adenocarcinoma and squamous cell carcinomas of the uterine cervix, anus, and lung.

    PubMed

    Chang, Alex; Amin, Ali; Gabrielson, Edward; Illei, Peter; Roden, Richard B; Sharma, Rajni; Epstein, Jonathan I

    2012-10-01

    Distinguishing invasive high-grade urothelial carcinoma (UC) from other carcinomas occurring in the genitourinary tract may be difficult. The differential diagnosis includes high-grade prostatic adenocarcinoma, spread from an anal squamous cell carcinoma (SCC), or spread from a uterine cervical SCC. In terms of metastatic UC, the most common problem is differentiating spread of UC to the lung from a primary pulmonary SCC. Immunohistochemical analysis (IHC) for GATA binding protein 3 (GATA3), thrombomodulin (THROMBO), and uroplakin III was performed on a tissue microarray (TMA) containing 35 cases of invasive high-grade UC. GATA3 IHC was also performed on TMAs containing 38 high-grade (Gleason score ≥8) prostatic adenocarcinomas, representative tissue sections from 15 invasive anal SCCs, representative tissue sections from 19 invasive cervical SCCs, and TMAs with 12 invasive cervical carcinomas of the cervix [SCC (n=10), SCC with neuroendocrine features (n=1), and adenosquamous carcinoma (n=1)]. In addition, GATA3 IHC was performed on representative tissue sections from 15 pulmonary UC metastases and a TMA with 25 SCCs of the lung and 5 pulmonary non-small cell carcinomas with squamous features. GATA3, THROMBO, and uroplakin III were positive in 28 (80%), 22 (63%), and 21 (60%) cases of high-grade UC, respectively. All cases of GATA3-positive staining were nonfocal; 25 (89%) cases demonstrated moderate to strong staining, and 3 (11%) demonstrated weak staining. Of the 7 cases that failed to express GATA3, 5 were positive for THROMBO and/or uroplakin III, whereas 2 were negative for all 3 markers. None of the 38 high-grade prostatic adenocarcinomas was positive for GATA3. Weak GATA3 staining was present in occasional basal cells of benign prostate glands, in a few benign atrophic glands, and in urothelial metaplasia. Of the 15 cases of anal SCCs, 2 (7%) cases showed focal weak staining, and 1 (3%) showed focal moderate staining. Weak staining was also rarely

  16. Reduced utilization of Man5GlcNAc2-P-P-lipid in a Lec9 mutant of Chinese hamster ovary cells: analysis of the steps in oligosaccharide-lipid assembly.

    PubMed

    Hall, C W; McLachlan, K R; Krag, S S; Robbins, A R

    1997-11-01

    Recently we reported that CHB11-1-3, a Chinese hamster ovary cell mutant defective in glycosylation of asparagine-linked proteins, is defective in the synthesis of dolichol [Quellhorst et al., 343:19-26, 1997: Arch Biochem Biophys]. CHB11-1-3 was found to be in the Lec9 complementation group, which synthesizes polyprenol rather than dolichol. In this paper, levels of various polyprenyl derivatives in CHB11-1-3 are compared to levels of the corresponding dolichyl derivatives in parental cells. CHB11-1-3 was found to maintain near normal levels of Man5GlcNAc2-P-P-polyprenol and mannosylphosphorylpolyprenol, despite reduced rates of synthesis, by utilizing those intermediates at a reduced rate. The Man5GlcNAc2 oligosaccharide attached to prenol in CHB11-1-3 cells and to dolichol in parental cells is the same structure, as determined by acetolysis. Man5GlcNAc2-P-P-polyprenol and Man5GlcNAc5-P-P-dolichol both appeared to be translocated efficiently in an in vitro reaction. Glycosylation of G protein was compared in vesicular stomatitus virus (VSV)-infected parent and mutant; although a portion of G protein was compared in vesicular stomatitus virus (VSV)-infected parent and mutant; although a portion of G protein was normally glycosylated in CHB11-1-3 cells, a large portion of G was underglycosylated, resulting in the addition of either one or no oligosaccharide to G. Addition of a single oligosaccharide occurred randomly rather than preferentially at one of the two sites.

  17. Instructional Facility Utilization.

    ERIC Educational Resources Information Center

    Kalamazoo Valley Community Coll., MI.

    Data describing campus facility use for instructional and related purposes for one week of activity in Fall 1978 were collected and evaluated at Kalamazoo Valley Community College. Four measures of space utilization were used: (1) percent of available time used; (2) percent of available space used; (3) percent of scheduled space utilized; and (4)…

  18. Teuchos Utility Package

    2004-03-01

    Teuchos is designed to provide portable, object-oriented tools for Trillnos developers and users. This includes templated wrappers to BLAS/LAPACK, a serial dense matrix class, a parameter list, XML parsing utilities, reference counted pointer (smart pointer) utilities, and more. These tools are designed to run on both serial and parallel computers.

  19. Tribal water utility management

    SciTech Connect

    Not Available

    1993-03-01

    Contents: primacy program (what is primacy, advantages and disadvantages, treatment as a state, grant applications and funding); safe drinking water act (sampling requirements, coliform standard, public notification, surface water treatment rule impacts, uic and wellhead protection programs, lead/copper rule); water utility management (how is the utility program evaluated, who's responsible, what is the board and tribal council role).

  20. Quantitative detection of the tumor-associated antigen large external antigen in colorectal cancer tissues and cells using quantum dot probe.

    PubMed

    Wang, Shuo; Li, Wanming; Yuan, Dezheng; Song, Jindan; Fang, Jin

    2016-01-01

    The large external antigen (LEA) is a cell surface glycoprotein that has been proven to be highly expressed in colorectal cancer (CRC) as a tumor-associated antigen. To evaluate and validate the relationship between LEA expression and clinical characteristics of CRC with high efficiency, LEA expression levels were detected in 85 tissue blocks from CRC patients by quantum dot-based immunohistochemistry (QD-IHC) combined with imaging quantitative analysis using quantum dots with a 605 nm emission wavelength (QD605) conjugated to an ND-1 monoclonal antibody against LEA as a probe. Conventional IHC was performed in parallel for comparison. Both QD-IHC and conventional IHC showed that LEA was specifically expressed in CRC, but not in non-CRC tissues, and high LEA expression was significantly associated with a more advanced T-stage (P<0.05), indicating that LEA is likely to serve as a CRC prognostic marker. Compared with conventional IHC, receiver operating characteristic analysis revealed that QD-IHC possessed higher sensitivity, resulting in an increased positive detection rate of CRC, from 70.1% to 89.6%. In addition, a simpler operation, objective analysis of results, and excellent repeatability make QD-IHC an attractive alternative to conventional IHC in clinical practice. Furthermore, to explore whether the QD probes can be utilized to quantitatively detect living cells or single cells, quantum dot-based immunocytochemistry (QD-ICC) combined with imaging quantitative analysis was developed to evaluate LEA expression in several CRC cell lines. It was demonstrated that QD-ICC could also predict the correlation between LEA expression and the T-stage characteristics of the cell lines, which was confirmed by flow cytometry. The results of this study indicate that QD-ICC has the potential to noninvasively detect rare circulating tumor cells in clinical samples in real clinical applications.

  1. NMDA receptors are expressed in human ovarian cancer tissues and human ovarian cancer cell lines

    PubMed Central

    North, William G; Liu, Fuli; Tian, Ruiyang; Abbasi, Hamza; Akerman, Bonnie

    2015-01-01

    We have earlier demonstrated that breast cancer and small-cell lung cancer express functional NMDA receptors that can be targeted to promote cancer cell death. Human ovarian cancer tissues and human ovarian cancer cell lines (SKOV3, A2008, and A2780) have now been shown to also express NMDA-receptor subunit 1 (GluN1) and subunit 2B (GluN2B). Seventeen ovarian cancers in two arrays were screened by immunohistochemistry using polyclonal antibodies that recognize an extracellular moiety on GluN1 and on GluN2B. These specimens comprised malignant tissue with pathology diagnoses of serous papillary cystadenocarcinoma, endometrioid adenocarcinoma, and clear-cell carcinoma. Additionally, archival tissues defined as ovarian adenocarcinoma from ten patients treated at this institute were also evaluated. All of the cancerous tissues demonstrated positive staining patterns with the NMDA-receptor antibodies, while no staining was found for tumor-adjacent normal tissues or sections of normal ovarian tissue. Human ovarian adenocarcinoma cell lines (A2008, A2780, SKOV3) were demonstrated to express GluN1 by Western blotting, but displayed different levels of expression. Through immunocytochemistry utilizing GluN1 antibodies and imaging using a confocal microscope, we were able to demonstrate that GluN1 protein is expressed on the surface of these cells. In addition to these findings, GluN2B protein was demonstrated to be expressed using polyclonal antibodies against this protein. Treatment of all ovarian cell lines with antibodies against GluN1 was found to result in decreased cell viability (P<0.001), with decreases to 10%–25% that of untreated cells. Treatment of control HEK293 cells with various dilutions of GluN1 antibodies had no effect on cell viability. The GluN1 antagonist MK-801 (dizocilpine maleate) and the GluN2B antagonist ifenprodil, like antibodies, dramatically decreased the viability of A2780 ovarian tumor cells (P<0.01). Treatment of A2780 tumor xenografts with

  2. NMDA receptors are expressed in human ovarian cancer tissues and human ovarian cancer cell lines.

    PubMed

    North, William G; Liu, Fuli; Tian, Ruiyang; Abbasi, Hamza; Akerman, Bonnie

    2015-01-01

    We have earlier demonstrated that breast cancer and small-cell lung cancer express functional NMDA receptors that can be targeted to promote cancer cell death. Human ovarian cancer tissues and human ovarian cancer cell lines (SKOV3, A2008, and A2780) have now been shown to also express NMDA-receptor subunit 1 (GluN1) and subunit 2B (GluN2B). Seventeen ovarian cancers in two arrays were screened by immunohistochemistry using polyclonal antibodies that recognize an extracellular moiety on GluN1 and on GluN2B. These specimens comprised malignant tissue with pathology diagnoses of serous papillary cystadenocarcinoma, endometrioid adenocarcinoma, and clear-cell carcinoma. Additionally, archival tissues defined as ovarian adenocarcinoma from ten patients treated at this institute were also evaluated. All of the cancerous tissues demonstrated positive staining patterns with the NMDA-receptor antibodies, while no staining was found for tumor-adjacent normal tissues or sections of normal ovarian tissue. Human ovarian adenocarcinoma cell lines (A2008, A2780, SKOV3) were demonstrated to express GluN1 by Western blotting, but displayed different levels of expression. Through immunocytochemistry utilizing GluN1 antibodies and imaging using a confocal microscope, we were able to demonstrate that GluN1 protein is expressed on the surface of these cells. In addition to these findings, GluN2B protein was demonstrated to be expressed using polyclonal antibodies against this protein. Treatment of all ovarian cell lines with antibodies against GluN1 was found to result in decreased cell viability (P<0.001), with decreases to 10%-25% that of untreated cells. Treatment of control HEK293 cells with various dilutions of GluN1 antibodies had no effect on cell viability. The GluN1 antagonist MK-801 (dizocilpine maleate) and the GluN2B antagonist ifenprodil, like antibodies, dramatically decreased the viability of A2780 ovarian tumor cells (P<0.01). Treatment of A2780 tumor xenografts with

  3. Highly Efficient Neural Differentiation of CD34-Positive Hair-Follicle-Associated Pluripotent Stem Cells Induced by Retinoic Acid and Serum-Free Medium.

    PubMed

    Sagha, Mohsen; Najafzadeh, Nowruz

    2016-01-01

    Neural differentiation of hair-follicle-associated pluripotent (HAP) stem cells residing in the bulge area is a promising autologous source for stem cell therapy. In the present chapter, we describe the identification and enrichment of CD34(+) HAP stem cells by magnetic-activated cell sorting (MACS), and induce them to differentiate into neuronal and glial cells using defined neural-induction media. The different neural cell populations arising during in vitro differentiation from HAP stem cells are characterized by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry assay. PMID:27431256

  4. Utility of the omentum in sacral reconstruction following total sacrectomy due to recurrent and irradiated giant cell tumour of the spine.

    PubMed

    Unal, Cigdem; Eren, Guler Gamze; Isil, Eda; Alponat, Ahmet; Sarlak, Ahmet

    2012-01-01

    Reconstruction of the lumbosacral region after surgical excision of irradiated and recurrent spinal giant cell tumours remains a challenging problem. In this case report, we describe the use of the pedicled omentum flap in reconstruction of an irradiated and infected wide sacral defect of a 19-year-old male patient. The patient had radiotherapy and subsequent wide surgical resection after recurrence of the tumour. A myocutaneous flap from the gluteal area had failed previously. Local flap options could not be used because of the recent radiotherapy to the gluteal area. Since the patient had a laparotomy for tumour resection and a colostomy, abdominal muscles were not considered reliable for reconstructive procedures. A pedicled omentum flap was chosen as a reconstructive option because of its rich blood supply, large surface area, and angiogenic capacity. This report aims to describe the use of the pedicled omentum flap for reconstruction of the lumbosacral area following surgical resection of a spinal tumour, when gluteal and abdominal flap options for reconstruction are jeopardised.

  5. The impact of brain metastasis on quality of life, resource utilization and survival in patients with non-small-cell lung cancer.

    PubMed

    Peters, Solange; Bexelius, Christin; Munk, Veronica; Leighl, Natasha

    2016-04-01

    This systematic review aims to improve understanding of the burden of disease associated with brain metastases from non-small-cell lung cancer (NSCLC) in terms of survival, quality of life (QoL) and economic impact. PubMed/MEDLINE, Cochrane collaboration and EMBASE databases were searched for articles published in English from 2000 to 2014. Of 3288 abstracts retrieved, 3156 were eliminated without a full-text review. Of the 132 articles that received a full-text review, a final set of 93 articles was included in an initial literature analysis. In order to homogenize the patient populations evaluated, we included entries that were either entirely composed of NSCLC patients or that had >50% of NSCLC patients in the total study population. From the studies identified in this systematic review, median OS and PFS varied based on the type of treatment received, although whole-brain radiotherapy (WBRT) was associated with the shortest OS and PFS durations. Regimens incorporating targeted therapy in molecularly selected patients were associated with the longest OS and PFS durations. QoL findings varied among studies, generally WBRT resulted in stable or worsening QoL scores rather than improvements. Healthcare costs were increased following diagnosis of brain metastases regardless of treatment. The findings from this review highlight the need for more effective treatments of brain metastases from NSCLC that improve survival function, QoL and potentially decrease costs. PMID:27019457

  6. The impact of brain metastasis on quality of life, resource utilization and survival in patients with non-small-cell lung cancer.

    PubMed

    Peters, Solange; Bexelius, Christin; Munk, Veronica; Leighl, Natasha

    2016-04-01

    This systematic review aims to improve understanding of the burden of disease associated with brain metastases from non-small-cell lung cancer (NSCLC) in terms of survival, quality of life (QoL) and economic impact. PubMed/MEDLINE, Cochrane collaboration and EMBASE databases were searched for articles published in English from 2000 to 2014. Of 3288 abstracts retrieved, 3156 were eliminated without a full-text review. Of the 132 articles that received a full-text review, a final set of 93 articles was included in an initial literature analysis. In order to homogenize the patient populations evaluated, we included entries that were either entirely composed of NSCLC patients or that had >50% of NSCLC patients in the total study population. From the studies identified in this systematic review, median OS and PFS varied based on the type of treatment received, although whole-brain radiotherapy (WBRT) was associated with the shortest OS and PFS durations. Regimens incorporating targeted therapy in molecularly selected patients were associated with the longest OS and PFS durations. QoL findings varied among studies, generally WBRT resulted in stable or worsening QoL scores rather than improvements. Healthcare costs were increased following diagnosis of brain metastases regardless of treatment. The findings from this review highlight the need for more effective treatments of brain metastases from NSCLC that improve survival function, QoL and potentially decrease costs.

  7. Exploring 1,4-dihydroxyanthraquinone as long-range emissive ratiometric fluorescent probe for signaling Zn(2+)/PO4(3-): Ensemble utilization for live cell imaging.

    PubMed

    Sinha, Sougata; Gaur, Pankaj; Mukherjee, Trinetra; Mukhopadhyay, Subhrakanti; Ghosh, Subrata

    2015-07-01

    Fluorescent 1,4-dihydroxyanthraquinone 1 was found to demonstrate its ratiometric signaling property upon interaction with divalent zinc (Zn(2+)). While the probe itself exhibited fluorescence emission in the yellow region (λem=544 nm and 567 nm), binding with Zn(2+) induced strong emission in the orange region (λem=600 nm) which was mainly due to a combination of CHEF and ICT mechanism. The probe was found to be highly sensitive toward the detection of zinc and the limit of detection (LOD) was calculated to be 9×10(-7) M. The possibility of using this probe for real-time analysis was strongly supported by the striking stability of fluorescence signal for more than five days with similar fluorescence intensity as observed during instant signaling. The present probe works within physiological pH range and is devoid of any interference caused by the same group elements such as Cd(2+)/Hg(2+). The probe possesses excellent excitation/emission wavelength profile and can penetrate cell membrane to image low concentration of zing inside living system. The in situ formed zinc-probe ensemble was further explored as ratiometric sensing platform for detecting another bio-relevant analyte phosphate anion through a zinc-displacement approach.

  8. Evaluation of the utility of subjective clinical parameters for estimating fecal egg counts and packed cell volume in Canadian sheep flocks.

    PubMed

    Mederos, A; Kelton, D; Peregrine, A S; VanLeeuwen, J; Fernández, S; LeBoeuf, A; Menzies, P; Martin, R

    2014-10-15

    A study was conducted in sheep on Canadian farms to describe the relationship between packed cell volume (PCV) or fecal egg counts (FEC) and subjective clinical parameters that may indicate the severity of parasitic gastroenteritis. Twenty-one farms in Ontario (ON) and 8 farms in Quebec (QC) were purposively selected and visited during April-May (spring) and August (summer) 2007. At each farm visit, blood and fecal samples were collected from 10 ewes and 10 female lambs; body condition score (BCS), dag score (DS), fecal consistency score (FCS) and FAMACHA score were recorded for all sampled sheep. Packed cell volume was determined for all blood samples, and FEC were performed for all fecal samples. Summary statistics and simple correlations were performed for the parameters recorded. Two mixed models with random effects at the farm level were developed; one using PCV as the response variable and another using the natural log of eggs per gram of feces (lnEPG). Finally, the residuals from both models were correlated to the covariates in the models. The mean PCV values during the spring were 29.7% and 36.7% for lambs, and 28.8% and 31.1% for ewes, in ON and QC, respectively. During the summer, the mean PCV was 32.0% and 32.8% for lambs, and 30.1% and 29.9% for ewes, in ON and QC, respectively. The arithmetic mean FEC per gram of feces (EPG) during the spring was 3 and 2 for lambs, and 1266 and 789 for ewes, in ON and QC, respectively, whereas during summer the arithmetic mean EPG was 907 and 237 for lambs, and 458 and 246 for ewes, in ON and QC, respectively. Results from simple correlations indicated that PCV was negatively correlated with lnEPG (r = -0.255; r(2) = 6.5%) and FAMACHA (r = -0.312; r(2) = 9.7%), and positively correlated with BCS (r = 0.317; r(2) = 10%). LnEPG was negatively correlated with BCS (r = -0.232; r(2) = 5.4%) and PCV (r = -0.255; r(2) = 6.5%), but positively correlated with FAMACHA (r = 0.178; r(2) = 3.2%) and DS (r = 0.086; r(2) = 0

  9. Utility of the tourniquet test and the white blood cell count to differentiate dengue among acute febrile illnesses in the emergency room.

    PubMed

    Gregory, Christopher J; Lorenzi, Olga D; Colón, Lisandra; García, Arleene Sepúlveda; Santiago, Luis M; Rivera, Ramón Cruz; Bermúdez, Liv Jossette Cuyar; Báez, Fernando Ortiz; Aponte, Delanor Vázquez; Tomashek, Kay M; Gutierrez, Jorge; Alvarado, Luisa

    2011-12-01

    Dengue often presents with non-specific clinical signs, and given the current paucity of accurate, rapid diagnostic laboratory tests, identifying easily obtainable bedside markers of dengue remains a priority. Previous studies in febrile Asian children have suggested that the combination of a positive tourniquet test (TT) and leucopenia can distinguish dengue from other febrile illnesses, but little data exists on the usefulness of these tests in adults or in the Americas. We evaluated the diagnostic accuracy of the TT and leucopenia (white blood cell count <5000/mm(3)) in identifying dengue as part of an acute febrile illness (AFI) surveillance study conducted in the Emergency Department of Saint Luke's Hospital in Ponce, Puerto Rico. From September to December 2009, 284 patients presenting to the ED with fever for 2-7 days and no identified source were enrolled. Participants were tested for influenza, dengue, leptospirosis and enteroviruses. Thirty-three (12%) patients were confirmed as having dengue; 2 had dengue co-infection with influenza and leptospirosis, respectively. An infectious etiology was determined for 141 others (136 influenza, 3 enterovirus, 2 urinary tract infections), and 110 patients had no infectious etiology identified. Fifty-two percent of laboratory-positive dengue cases had a positive TT versus 18% of patients without dengue (P<0.001), 87% of dengue cases compared to 28% of non-dengue cases had leucopenia (P<0.001). The presence of either a positive TT or leucopenia correctly identified 94% of dengue patients. The specificity and positive predictive values of these tests was significantly higher in the subset of patients without pandemic influenza A H1N1, suggesting improved discriminatory performance of these tests in the absence of concurrent dengue and influenza outbreaks. However, even during simultaneous AFI outbreaks, the absence of leucopenia combined with a negative tourniquet test may be useful to rule out dengue.

  10. Gender, Race, and Survival: A Study in Non-Small-Cell Lung Cancer Brain Metastases Patients Utilizing the Radiation Therapy Oncology Group Recursive Partitioning Analysis Classification

    SciTech Connect

    Videtic, Gregory M.M.; Reddy, Chandana A.; Chao, Samuel T.; Rice, Thomas W.; Adelstein, David J.; Barnett, Gene H.; Mekhail, Tarek M.; Vogelbaum, Michael A.; Suh, John H.

    2009-11-15

    Purpose: To explore whether gender and race influence survival in non-small-cell lung cancer (NSCLC) in patients with brain metastases, using our large single-institution brain tumor database and the Radiation Therapy Oncology Group recursive partitioning analysis (RPA) brain metastases classification. Methods and materials: A retrospective review of a single-institution brain metastasis database for the interval January 1982 to September 2004 yielded 835 NSCLC patients with brain metastases for analysis. Patient subsets based on combinations of gender, race, and RPA class were then analyzed for survival differences. Results: Median follow-up was 5.4 months (range, 0-122.9 months). There were 485 male patients (M) (58.4%) and 346 female patients (F) (41.6%). Of the 828 evaluable patients (99%), 143 (17%) were black/African American (B) and 685 (83%) were white/Caucasian (W). Median survival time (MST) from time of brain metastasis diagnosis for all patients was 5.8 months. Median survival time by gender (F vs. M) and race (W vs. B) was 6.3 months vs. 5.5 months (p = 0.013) and 6.0 months vs. 5.2 months (p = 0.08), respectively. For patients stratified by RPA class, gender, and race, MST significantly favored BFs over BMs in Class II: 11.2 months vs. 4.6 months (p = 0.021). On multivariable analysis, significant variables were gender (p = 0.041, relative risk [RR] 0.83) and RPA class (p < 0.0001, RR 0.28 for I vs. III; p < 0.0001, RR 0.51 for II vs. III) but not race. Conclusions: Gender significantly influences NSCLC brain metastasis survival. Race trended to significance in overall survival but was not significant on multivariable analysis. Multivariable analysis identified gender and RPA classification as significant variables with respect to survival.

  11. Utility of the Tourniquet Test and the White Blood Cell Count to Differentiate Dengue among Acute Febrile Illnesses in the Emergency Room

    PubMed Central

    Gregory, Christopher J.; Lorenzi, Olga D.; Colón, Lisandra; Sepúlveda García, Arleene; Santiago, Luis M.; Cruz Rivera, Ramón; Cuyar Bermúdez, Liv Jossette; Ortiz Báez, Fernando; Vázquez Aponte, Delanor; Tomashek, Kay M.; Gutierrez, Jorge; Alvarado, Luisa

    2011-01-01

    Dengue often presents with non-specific clinical signs, and given the current paucity of accurate, rapid diagnostic laboratory tests, identifying easily obtainable bedside markers of dengue remains a priority. Previous studies in febrile Asian children have suggested that the combination of a positive tourniquet test (TT) and leucopenia can distinguish dengue from other febrile illnesses, but little data exists on the usefulness of these tests in adults or in the Americas. We evaluated the diagnostic accuracy of the TT and leucopenia (white blood cell count <5000/mm3) in identifying dengue as part of an acute febrile illness (AFI) surveillance study conducted in the Emergency Department of Saint Luke's Hospital in Ponce, Puerto Rico. From September to December 2009, 284 patients presenting to the ED with fever for 2–7 days and no identified source were enrolled. Participants were tested for influenza, dengue, leptospirosis and enteroviruses. Thirty-three (12%) patients were confirmed as having dengue; 2 had dengue co-infection with influenza and leptospirosis, respectively. An infectious etiology was determined for 141 others (136 influenza, 3 enterovirus, 2 urinary tract infections), and 110 patients had no infectious etiology identified. Fifty-two percent of laboratory-positive dengue cases had a positive TT versus 18% of patients without dengue (P<0.001), 87% of dengue cases compared to 28% of non-dengue cases had leucopenia (P<0.001). The presence of either a positive TT or leucopenia correctly identified 94% of dengue patients. The specificity and positive predictive values of these tests was significantly higher in the subset of patients without pandemic influenza A H1N1, suggesting improved discriminatory performance of these tests in the absence of concurrent dengue and influenza outbreaks. However, even during simultaneous AFI outbreaks, the absence of leucopenia combined with a negative tourniquet test may be useful to rule out dengue. PMID:22163057

  12. Robert Feulgen Prize Lecture 1999. Detection and amplification systems for sensitive, multiple-target DNA and RNA in situ hybridization: looking inside cells with a spectrum of colors.

    PubMed

    Speel, E J

    1999-08-01

    In situ hybridization (ISH) is a powerful technique for localizing specific nucleic acid sequences (DNA, RNA) in microscopic preparations of tissues, cells, chromosomes, and linear DNA fibers. To date, a wide variety of research and diagnostic applications of ISH have been described, making the technique an integral part of studies concerning gene mapping, gene expression, RNA processing and transport, the three-dimensional organization of the nucleus, tumor genetics, microbial infections, and prenatal diagnosis. In this review, I first describe the ISH procedure in short and then focus on the currently available non-radioactive probe-labeling and cytochemical detection methodologies that are utilized to visualize one or multiple different nucleic acid targets in situ with different colors. Special emphasis is placed on the procedures applying fluorescence and brightfield microscopy, the simultaneous detection of nucleic acids and proteins by combined ISH and immunocytochemistry, and, in addition, on the recent progress that has been made with the introduction of signal amplification procedures to increase the detection sensitivity of ISH. Finally, a comparison of fluorescence, enzyme cytochemical, and colloidal gold silver probe detection systems will be presented, and possible future directions of in situ nucleic acid detection will be discussed.

  13. Robert Feulgen Prize Lecture 1999. Detection and amplification systems for sensitive, multiple-target DNA and RNA in situ hybridization: looking inside cells with a spectrum of colors.

    PubMed

    Speel, E J

    1999-08-01

    In situ hybridization (ISH) is a powerful technique for localizing specific nucleic acid sequences (DNA, RNA) in microscopic preparations of tissues, cells, chromosomes, and linear DNA fibers. To date, a wide variety of research and diagnostic applications of ISH have been described, making the technique an integral part of studies concerning gene mapping, gene expression, RNA processing and transport, the three-dimensional organization of the nucleus, tumor genetics, microbial infections, and prenatal diagnosis. In this review, I first describe the ISH procedure in short and then focus on the currently available non-radioactive probe-labeling and cytochemical detection methodologies that are utilized to visualize one or multiple different nucleic acid targets in situ with different colors. Special emphasis is placed on the procedures applying fluorescence and brightfield microscopy, the simultaneous detection of nucleic acids and proteins by combined ISH and immunocytochemistry, and, in addition, on the recent progress that has been made with the introduction of signal amplification procedures to increase the detection sensitivity of ISH. Finally, a comparison of fluorescence, enzyme cytochemical, and colloidal gold silver probe detection systems will be presented, and possible future directions of in situ nucleic acid detection will be discussed. PMID:10460463

  14. Benzo[ghi]perylene activates the AHR pathway to exert biological effects on the NL-20 human bronchial cell line.

    PubMed

    Zaragoza-Ojeda, Montserrat; Eguía-Aguilar, Pilar; Perezpeña-Díazconti, Mario; Arenas-Huertero, Francisco

    2016-08-10

    Polycyclic aromatic hydrocarbons (PAH) are produced by incomplete combustion of organic material. In the Mexico City atmosphere, the most abundant PAH is benzo[ghi]perylene (BghiP), a gasoline combustion marker. At present, there are no reports of the effects of BghiP on human bronchial cells, so the aim of the study was to evaluate the effects in vitro of BghiP on the NL-20 cell line. Results showed that BghiP induced the formation of small vesicles throughout the cytoplasm, with absence of nuclear fragmentation. At 48h exposition, damage in cell membrane increased significantly at 1.24μg/mL of BghiP (p<0.05). Immunocytochemistry revealed that BghiP provokes nuclear translocation of AhR receptor, which indicates that this compound can induce transcription of genes via receptor binding (AhR pathway activation). BghiP induced a two-fold increase (p<0.05) in the expression of AhR and CYP4B1 (a lung-specific pathway effector). In the presence of the receptor antagonist CH-223191, the loss of viability, the nuclear translocation and the overexpression of genes decreased, though this did not prevent the formation of vesicles. BghiP induced oxidative stress and in presence of the receptor antagonist this increased significantly. In conclusion, BghiP can activate the overexpression of AhR and CYP4B1, and the effects are abated by the AhR receptor antagonist. This is the first report to prove that BghiP utilizes the AhR pathway to exert its toxic effects on the NL-20 human bronchial cell line . PMID:27234499

  15. STARD-compliant article: The utility of red cell distribution width to predict mortality for septic patients visiting the emergency department.

    PubMed

    Chen, Chun-Kuei; Lin, Shen-Che; Wu, Chin-Chieh; Chen, Li-Min; Tzeng, I-Shiang; Chen, Kuan-Fu

    2016-06-01

    Sepsis is a common condition in the emergency department that is associated with high mortality. Red blood cell distribution width (RDW) has been used as a simple prognosis predictor for patients with community-acquired pneumonia, gram-negative bacteremia, and severe sepsis or septic shock. To evaluate the performance of RDW to predict in-hospital mortality among septic patients, we conducted a hospital-based retrospective cohort study in an emergency department of a tertiary teaching hospital. RDW was compared with other commonly used clinical prediction scores (Systemic Inflammatory Response Syndrome (SIRS), Mortality in Emergency Department Sepsis (MEDS) and the Confusion, Urea nitrogen, Respiratory rate, Blood pressure, 65 years of age and older (CURB65)). Of 6973 consecutive adult patients with a clinical diagnosis of sepsis and 2 sets of blood culture ordered by physicians, 477 (6.8%) died. The mortality group had higher RDW levels than the survival group (15.7% vs 13.8%). After dividing RDW into quartiles, the patients in the highest RDW quartile (RDW >15.6%; mortality, 16.7%) had more than twice the risk of in-hospital mortality compared with patients in the second highest quartile (RDW >14% and <15.6%; mortality, 7.3%), whereas the mortality rate in the lowest RDW quartile (<13.1%) was only 1.6%. The area under the receiver operating characteristic curve of RDW to predict mortality was 0.75 (95% confidence interval, 0.72-0.77), which is significantly higher than the areas under the curve of clinical prediction rules (SIRS, MEDS, and CURB65). After integrating RDW into these scores, all scores performed better in predicting mortality (0.73, 0.72, and 0.77, for SIRS, MEDS, and CURB65, respectively). RDW could be an independent predictor of mortality among septic patients. Clinicians could classify the septic patients into different risk groups according to RDW quartiles. For more accurate mortality prediction, RDW could be a potential parameter to be

  16. Device and Circuit Modeling and Development of a Non-Volatile Random Access Memory Cell, Utilizing AN Amorphous Silicon Thin-Film Floating-Gate Transistor Based Technology.

    NASA Astrophysics Data System (ADS)

    Riggio, Salvatore Richard, Jr.

    1994-01-01

    High density storage mechanisms are generally created using either magnetic or optical implementation techniques. Both of these techniques require mechanical transport of the medium and, therefore, have low reliability factors. These devices also generate unwanted low level ambient noise, which is of particular concern when considering modern quiet office standards. Additionally, optical techniques tend to be read-only in nature. Both mechanisms exhibit random access times that are measured in milli-seconds, rather than in micro-seconds. Therefore, the creation of a non-volatile random access memory as a replacement for the above mentioned storage techniques would be of great advantage in terms of access time, reliability, and ambient noise level. Described within are the device and circuit modeling and fabrication techniques used to develop a non-volatile random access memory cell from an amorphous silicon thin -film transistor based technology. Amorphous silicon thin-film transistors are fabricated by depositing the metal, the insulator and the semiconductor materials with a sputtering mechanism in a vacuum at 220 degrees centigrade, rather than by diffusion at 2000 degrees centigrade, as is done with crystalline silicon. By depositing a metal in the insulator, which is located between the gate and the channel, and by using an insulator material with extremely high resistivity, one can store charge in the gate region for a long period of time without external power. For example, this period of time can be as little as one week or as long as over one year. With a periodic refresh, one can extend the memory time of this storage mechanism indefinitely. Thin-film transistors can be deposited on a variety of materials such as glass, quartz or plastic by means of a stationary or continuous motion fabrication system. This material can be either rigid or flexible, and can be comparatively large in size. This allows for much greater circuit de