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Sample records for cells utilizing immunocytochemistry

  1. Digital microfluidic immunocytochemistry in single cells

    PubMed Central

    Ng, Alphonsus H. C.; Chamberlain, M. Dean; 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. [Distribution and accumulation of antibiotics in cells and tissues and toxicity studies by immunocytochemistry].

    PubMed

    Fujiwara, Kunio

    2011-01-01

    No true immunocytochemistry (ICC) for drugs nor its application to pharmacokinetic studies is available. Recently, our studies have shown that ICC for drugs is extremely useful for such studies by utilizing easy and safe techniques, and gives direct evidence of drug localization. We have therefore developed antibodies and a series of pretreatment conditions for the immunodetection of drugs and have localized sites of drug uptake or accumulation in several tissues of rats following the administration of drugs. This review describes preparation of anti-drug antibody, specificity of antibody, fixation of drug in situ in rat tissues and cells, treatment of paraffin section specimens prior to immunoreaction, precision, and their application to a variety of types of antibiotics anti-cancer anthracyclines daunorubicin, doxorubicin, and epirubicin, bleomycin analog peplomycin, antimicrobial agents gentamicin, and amoxicillin. ICC for the anti-cancer anthracyclines demonstrated that the drug accumulates in a characteristic pattern in the heart, liver, kidney, gastrointestinal tract, and hair follicles, which represent the sites targeted by the drug toxicity. Some, but not all, of these drug accumulations are associated with the induction of apoptosis. It was also noted that there are striking differences in accumulation among the anthracyclines in rat tissues, maybe contributing the mechanisms of the differences in anti-tumor activities of the anthracyclines. Both ICCs for gentamicin and peplomycin identified characteristic necrotic-like cells in the specific sites of the kidney, suggesting the sites are readily affected by some chemotherapeutic agents. ICC for amoxicillin demonstrated that the sites of the drug accumulation in small intestine, liver and kidney are closely correlated with the specific sites in which certain transporter systems for penicillin occur. Thus, an ICC method is a potential new tool for pharmacokinetic studies of wide variety types of drugs

  3. Covalent affinity labeling, radioautography, and immunocytochemistry localize the glucocorticoid receptor in rat testicular Leydig cells

    SciTech Connect

    Stalker, A.; Hermo, L.; Antakly, T. )

    1989-12-01

    The presence and distribution of glucocorticoid receptors in the rat testis were examined by using 2 approaches: in vivo quantitative radioautography and immunocytochemistry. Radioautographic localization was made possible through the availability of a glucocorticoid receptor affinity label, dexamethasone 21-mesylate, which binds covalently to the glucocorticoid receptor, thereby preventing dissociation of the steroid-receptor complex. Adrenalectomized adult rats were injected with a tritiated (3H) form of this steroid into the testis and the tissue was processed for light-microscope radioautography. Silver grains were observed primarily over the Leydig cells of the interstitial space and to a lesser extent, over the cellular layers which make up the seminiferous epithelium, with no one cell type showing preferential labeling. To determine the specificity of the labeling, a 25- or 50-fold excess of unlabeled dexamethasone was injected simultaneously with the same dose of (3H)-dexamethasone 21-mesylate. In these control experiments, a marked reduction in label intensity was noted over the Leydig as well as tubular cells. Endocytic macrophages of the interstitium were non-specifically labeled, indicating uptake of the ligand possibly by fluid-phase endocytosis. A quantitative analysis of the label confirmed the presence of statistically significant numbers of specific binding sites for glucocorticoids in both Leydig cells and the cellular layers of the seminiferous epithelium; 86% of the label was found over Leydig cells, and only 14% over the cells of the seminiferous epithelium. These binding data were confirmed by light-microscope immunocytochemistry using a monoclonal antibody to the glucocorticoid receptor.

  4. Peritoneal fluid immunocytochemistry used for the diagnosis of a possible case of equine gastrointestinal B-cell lymphoma

    PubMed Central

    Duran, Maria Carolina; Starrak, Gregory; Dickinson, Ryan; Montgomery, Julia

    2016-01-01

    After physical examination, ultrasonographic evaluation of thorax and abdomen, and peritoneal fluid analysis, gastrointestinal neoplasia with suspected diffuse peritoneal metastasis was diagnosed in a 17-year-old Arabian gelding. The owner elected euthanasia and declined postmortem examination. Immunocytochemistry analysis of the peritoneal fluid resulted in a diagnosis of B-cell lymphoma. PMID:27247458

  5. Organization of ventrolateral periolivary cells of the cat superior olive as revealed by PEP-19 immunocytochemistry and Nissl stain.

    PubMed

    Spirou, G A; Berrebi, A S

    1996-04-22

    Ventrolateral periolivary cell groups, through their descending projections to the cochlear nucleus (CN) and local projections to principal nuclei of the superior olive, may participate in brainstem mechanisms mediating such tasks as signal detection in noisy environments and sound localization. Understanding the function of these cell groups can be improved by increased knowledge of the organization of their synaptic inputs in relation to their cellular characteristics. Immunocytochemistry for PEP-19 (a putative calcium binding protein) reveals four patterns of immunolabeling within the ventrolateral periolivary region. Three of the patterns, which have distinct fiber and punctate labeling characteristics, help to define three subdivisions of the lateral nucleus of the trapezoid body (LNTB). The fourth pattern defines two other nuclei, the anterolateral periolivary nucleus (rostral) and the posterior periolivary nucleus (caudal), which display many immunoreactive cell bodies but little fiber and punctate labeling. One of the subdivisions of the LNTB contains large PEP-19 immunolabeled puncta arranged in pericellular nests. Analysis of Nissl-stained sections reveals a neuronal population that resembles globular cells of the ventral cochlear nucleus (VCN) and which colocalizes with pericellular nests of large immunolabeled puncta. Cell counts reveal that roughly 10,000 neurons constitute the cat ventrolateral periolivary region, 9,000 of which are found in the LNTB. Three-dimensional reconstructions of auditory brainstem nuclei clarify the complex spatial relationships among these structures.

  6. Liquid-based cytology and cell block immunocytochemistry in veterinary medicine: comparison with standard cytology for the evaluation of canine lymphoid samples.

    PubMed

    Fernandes, N C C A; Guerra, J M; Réssio, R A; Wasques, D G; Etlinger-Colonelli, D; Lorente, S; Nogueira, E; Dagli, M L Z

    2016-08-01

    Liquid-based Cytology (LBC) consists of immediate wet cell fixation with automated slide preparation. We applied LBC, cell block (CB) and immunocytochemistry to diagnose canine lymphoma and compare results with conventional cytology. Samples from enlarged lymph nodes of 18 dogs were collected and fixed in preservative solution for automated slide preparation (LBC), CB inclusion and immunophenotyping. Two CB techniques were tested: fixed sediment method (FSM) and agar method (AM). Anti-CD79a, anti-Pax5, anti-CD3 and anti-Ki67 were used in immunocytochemistry. LBC smears showed better nuclear and nucleolar definition, without cell superposition, but presented smaller cell size and worse cytoplasmic definition. FSM showed consistent cellular groups and were employed for immunocytochemistry, while AM CBs presented sparse groups of lymphocytes, with compromised analysis. Anti-Pax-5 allowed B-cell identification, both in reactive and neoplastic lymph nodes. Our preliminary report suggests that LBC and FSM together may be promising tools to improve lymphoma diagnosis through fine-needle aspiration.

  7. [Comparative characteristic of the methods of immunocytochemistry and immunomagnetic separation for detection of cytokeratin-positive cells in bone marrow and lymph nodes].

    PubMed

    Lebedinskaia, O V; Shubina, I Zh; Donenko, F; Pirogov, A V; Kiselevskiĭ, M V

    2007-01-01

    Comparative study of cytokeratin-positive (CK+) cells, isolated from bone marrow and lymph node micrometastases in patients with lung and esophageal cancer, was performed using the methods of immunocytochemistry and immunomagnetic separation. It was found that immunocytochemical analysis permitted the detection of CK+ cells in the smears of bone marrow of oncologic patients. All the samples of bone marrow with immunocytochemically demonstrated CK+ cells, contained also the cellular complexes marked by magnetic spheres. The method of positive immunomagnetic separation has some advantages for the intraoperative diagnosis. However, when either method was applied, among CK+ cells in bone marrow and lymph nodes some false-positive, questionable cellular forms and hemopoietic cells expressing epithelial markers were found. Theses results suggest that for verification of micrometastases in these organs, an additional study using the standard staining of the slides, is required.

  8. Exocytosis sensitivity to growth hormone-releasing hormone in subsets of GH cells in rats under different corticosterone conditions. Ultrastructural study using microwave irradiation for fixation and immunocytochemistry.

    PubMed

    Ozawa, Hitoshi; Han, Fang; Kawata, Mitsuhiro

    2004-12-01

    Growth hormone (GH) cells in the rat anterior pituitary have been morphologically classified into three subtypes: type I (mature) containing large secretory granules about 350 nm in diameter, type II (intermediate) containing a mixture of large and small granules, and type III (immature) containing small granules about 150 nm in diameter. However, the functional implications of morphological heterogeneity, especially the different sensitivities to growth hormone-releasing hormone (GRH) under different corticosteroid conditions have not been elucidated to date. In the present study, by application of microwave irradiation (MWI) for fixation and immunocytochemistry, new findings of the exocytotic response have been revealed among the subsets of GH cells following adrenalectomy (ADX), corticosterone treatment and/or GRH treatment. The MWI gave effective results for fixation, especially for the permeability of the fixative, and showed good results for immunoelectron microscopy using the protein-A gold method. Moreover, the use of MWI greatly shortened the fixation, processing and immunolabeling times without compromising the quality of ultrastructural preservation and the specificity of labeling. The number of exocytotic figures was low in all subtypes of GH cells in the sham-operated control rats. GRH treatment induced a significant increase in exocytosis in each subtype of GH cells, particularly in type I (mature) and type II (intermediate) GH cells in the control rats. GRH injection to rats for 4 days after ADX also showed an increase in exocytosis, but the degree was significantly less in comparison with the GRH injection in the control group. Corticosterone replacement given to ADX rats induced a clear recovery of the exocytotic response to GRH to the control level. Serum GH content measured by radioimmunoassay correlated with these morphological results. These results suggest that the secretion of GH stimulated by GRH is closely related to corticosteroids, and

  9. Cell-specific expression of epithelial sodium channel alpha, beta, and gamma subunits in aldosterone-responsive epithelia from the rat: localization by in situ hybridization and immunocytochemistry

    PubMed Central

    1994-01-01

    A highly selective, amiloride-sensitive, epithelial sodium channel from rat colon (rENaC), composed of three homologous subunits termed alpha, beta, and gamma rENaC, has been cloned by functional expression and was proposed to mediate electrogenic sodium reabsorption in aldosterone- responsive epithelia. To determine whether rENaC could account for sodium absorption in vivo, we studied the cellular localization of the sodium channel messenger RNA subunits by in situ hybridization and their cellular and subcellular distribution by immunocytochemistry in the kidney, colon, salivary, and sweat glands of the rat. In the kidney, we show that the three subunit mRNAs are specifically co- expressed in the renal distal convoluted tubules (DCT), connecting tubules (CNT), cortical collecting ducts (CCD), and outer medullary collecting ducts (OMCD), but not in the inner medullary collecting ducts (IMCD). We demonstrate co-localization of alpha, beta, and gamma subunit proteins in the apical membrane of a majority of cells of CCD and OMCD. Our data indicate that alpha, beta, and gamma subunit mRNAs and proteins are co-expressed in the distal nephron (excepting IMCD), a localization that correlates with the previously described physiological expression of amiloride-sensitive electrogenic sodium transport. Our data, however, suggest that another sodium transport protein mediates electrogenic amiloride-sensitive sodium reabsorption in IMCD. We also localized rENaC to the surface epithelial cells of the distal colon and to the secretory ducts of the salivary gland and sweat gland, providing further evidence consistent with the hypothesis that the highly selective, amiloride-sensitive sodium channel is physiologically expressed in aldosterone-responsive cells. PMID:7806569

  10. 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

  11. Effective application of the methanol-based PreservCyt(™) fixative and the Cellient(™) automated cell block processor to diagnostic cytopathology, immunocytochemistry, and molecular biology.

    PubMed

    van Hemel, Bettien M; Suurmeijer, Albert J H

    2013-08-01

    We studied the feasibility of immunocytochemistry (ICC), in situ hybridization (ISH), and polymerase chain reaction (PCR) after Cellient(™) automated cell block processing, and tested whether methanol-based PreservCyt(™) fixation could replace formalin fixation, in an attempt to eliminate toxic formaldehyde vapors. Immunostaining with 30 different antibodies was performed on cell blocks from 73 FNA specimens and 42 body cavity fluid specimens prepared by Cellient(™) automated processing that uses the methanol-based fixative (PreservCyt(™) ). For each antibody we evaluated ICC in at least three different cell block specimens and compared it with immunohistochemistry (IHC) in formalin-fixed, paraffin-embedded (FFPE) histological sections from the corresponding tumors. The quality of DNA and RNA in Cellient(™) blocks was analyzed by ISH, applying a SYT gene break-apart assay and EBER probes, respectively. Moreover, DNA quality was analyzed by PCR by using primer sets for DNA products of 100, 200, 300, 400, 500, and 600 base pairs, and evaluated by gel electrophoresis. When compared with IHC results in corresponding FFPE tumor tissue from the same patient, 24 out of 30 antibodies showed concordant ICC results. With FISH, distinctive hybridization signals were observed for SYT DNA sequences and EB virus RNA sequences. With PCR, DNA products, up to 600 base pairs in size, were readily observed after gel electrophoresis. The antibodies that showed concordant immunostaining in Cellient(™) blocks could be applied to diagnostic algorithms that proved to be helpful in the discrimination of major tumor types (carcinoma, lymphoma, melanoma, and germ cell tumors), discrimination of carcinoma subtypes, and determination of primary tumor site in cases of metastatic carcinoma. In a separate study, we found that the application of ICC to this cell block technique provided additional diagnostic and clinically important information in 24% of 100 consecutive cases. The high

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

    PubMed

    Froment, Rémi; Gara-Boivin, Carolyn

    2015-12-01

    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.

  13. 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

  14. Improved method for combination of immunocytochemistry and Nissl staining.

    PubMed

    Kádár, Andrea; Wittmann, Gábor; Liposits, Zsolt; Fekete, Csaba

    2009-10-30

    Nissl staining is a widely used method to study morphology and pathology of neural tissue. After standard immunocytochemistry, the Nissl staining labels only the nucleus of neurons and the characteristic staining of the neuronal perikarya is absent or very weak. We hypothesized that the RNA degradation during the immunocytochemical treatment results in the loss of cytoplasmic staining with Nissl-dyes. To test this hypothesis, we used RNAse-free conditions for all steps of immunostaining. To further prevent the RNA-degradation by RNAse contaminations, the RNAse inhibitor heparin was added to all antibody-containing solutions. The efficiency of Nissl staining after standard and RNAse-free double-labeling immunocytochemistry was compared using antibodies against c-Fos and neuropeptide Y (NPY) on tissues of rats refed after 3 days of fasting. After standard immunocytochemistry, the Nissl-staining labeled the nuclei of neurons and only very faintly the cytoplasm of these cells. The RNAse-free treatment did not alter the distribution of immunoreaction signal, but preserved the staining of neuronal perikarya by the Nissl-dyes. In conclusion, the RNAse-free conditions during immunocytochemistry allow the labeling of neuronal perikarya by Nissl-dyes. The described method facilitates the mapping of immunocytochemical signals and makes possible the light microscopic examination of the innervation of neurons identified by their nuclear protein content.

  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. 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.

  17. Peripheral cell loss related to calcium binding protein immunocytochemistry in the dorsal cochlear nucleus in CBA/CaJ mice during aging.

    PubMed

    Idrizbegovic, E; Viberg, A; Bogdanovic, N; Canlon, B

    2001-01-01

    The influence of cochlear hair cell and spiral ganglia neuron loss on calcium binding protein immunoreactivity (calretinin, parvalbumin and calbindin) in the dorsal and posteroventral cochlear nuclei (DCN and PVCN) in CBA/CaJ (CBA) mice during aging (1-39 months) was determined. Since calcium binding proteins have buffering properties against calcium overload, they may have a protective role during aging. It is shown that the percentage of calretinin- and parvalbumin-immunopositive neurons in the DCN showed a statistically significant positive correlation with inner hair cell loss, outer hair cell loss, and spiral ganglion cell loss. A correlation was also found between aging and the auditory periphery, and calcium binding proteins in the DCN. These findings imply that the pathophysiological state of the auditory periphery may influence the neuronal homeostasis in the dorsal cochlear nucleus.

  18. p53, PCNA and Ki-67 expression in oral squamous cell carcinomas: the vagaries of fixation and microwave enhancement of immunocytochemistry.

    PubMed

    Allison, R T; Best, T

    1998-10-01

    Proliferation markers are widely used as indicators of tumour progression and aggression. Fixation and antigen retrieval methods may enhance the immunocytochemical sensitivity of these markers but may also lead to loss of specificity. As these methods are often used quantitatively, standardisation of internal and external methodology is paramount. This study aimed to compare the effects of alcohol and formalin fixation and of microwaving on the immunocytochemical demonstration of p53, PCNA and Ki-67 in oral squamous cell carcinoma using duplicate tissue blocks from 24 cases. Both qualitative and quantitative differences in antigen expression were revealed. Whilst alcohol fixation alone at least maintained and usually increased the strength of positive staining, microwaving alcohol-fixed sections often gave rise to non-specific staining. p53 staining following microwave enhancement of alcohol-fixed tissue showed a significant incidence of conversion of negative results to positive and of positive staining in unexpected tissue components. Alcohol fixation increased the sensitivity of PCNA detection with a far less dramatic loss of specificity. The results emphasise the need for careful standardisation of immunocytochemical methods, particularly when used quantitatively and for inter-laboratory comparisons.

  19. Fuel cells: A utilities perspective

    NASA Astrophysics Data System (ADS)

    Hessenius, Chris A.; Ang, Amos; Hamilton, Stephanie

    Southern California Edison (SCE) is actively assessing how to maximize the benefits from fuel cell power systems and other distributed generation (DG) technologies deployed along existing distribution level circuits. From a utility perspective, the viability of DG fuel cell systems increase as the technology matures and more "value-added" features are incorporated. As the number of DG projects grows in SCE's service territory and optimism increases about the potential uses, so does the need to better understand the impact wide-scale deployment may have on the performance of California's energy system. Understanding how DG technologies affect distribution level circuits and devising effective deployment strategies is essential for the technology to gain widespread acceptance and become an integral part of SCE's Transmission and Distribution (T&D) system planning. Simulation results are presented in this paper that indicate fuel cell systems combined with electronically switched power inverters capable or providing reactive power (a.k.a. VAR) support are more advantageous than fuel cell systems without such inverter features. In fact, for the SCE circuit analyzed, a strategically placed 2.5 MW fuel cell system with VAR support capabilities has a greater affect on circuit performance than a 3 MW fuel cell system without VAR support. Even though the 2.5 MW fuel cell system with VAR support inverter possesses 16.7% less power rating than the 3 MW system without VAR support, it was more effective in reducing circuit current flows, reducing distribution line losses, and maintaining circuit voltage within ±5% of 12.47 kilovolts (kV).

  20. Immunocytochemistry: an indispensable technique in routine cytology.

    PubMed

    Skoog, L; Tani, E

    2011-08-01

    Immunocytology is today accepted as an indispensable adjunct to cytomorphology. It has led to a dramatic increase in diagnostic accuracy and also allowed the identification of markers both for prognosis and targeted therapies. Most commercially available antibodies will perform in a reproducible and reliable way provided that the cytological specimen has been prepared and fixed properly. In this review various aspects of immunocytochemistry such as preparation of cytological specimens, fixation and choice of antibodies will be discussed. The specificity of the most commonly used antibodies is summarized and staining panels for various tumours are suggested. In addition, the use of markers for targeted therapy and theranostics is discussed, as well as a brief section on the identification of infectious agents.

  1. Neurochemical anatomy of the zebrafish retina as determined by immunocytochemistry.

    PubMed

    Yazulla, S; Studholme, K M

    2001-07-01

    The zebrafish retina is rapidly becoming a major preparation for the study of molecular genetic mechanisms underlying neural development and visual behavior. Studies utilizing retinal mutants would benefit by the availability of a data base on the distribution of neurotransmitter systems in the wild-type fish. To this end, the neurochemical anatomy of the zebrafish retina was surveyed by light microscopic immunocytochemistry. An extensive series of 60 separate antibodies were used to describe the distribution of major transmitter systems and a variety of neuron-associated membrane channels and proteins. These include markers (i.e., antibodies against enzymes, receptors, transporters) for transmitters: GABA, glycine, glutamate, biogenic amines, acetylcholine, cannabinoids and neuropeptides; as well as a sample of voltage-gated channels and synapse associated membrane proteins. Discussion of the comparative localization of these antibodies is restricted to other teleost fishes, particularly goldfish. Overall, there was great similarity in the distribution of the various markers, as might be expected. However, there were some notable differences, including several antibodies that did not label zebrafish at all, even though goldfish retinas that were processed in parallel, labeled beautifully. This survey is extensive, but not exhaustive, and hopefully will serve as a valuable resource for future studies of the zebrafish retina.

  2. 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

  3. Documentation of immunocytochemistry controls in the cytopathologic literature: a meta-analysis of 100 journal articles.

    PubMed

    Colasacco, Carol; Mount, Sharon; Leiman, Gladwyn

    2011-04-01

    Although a detailed description of the procedure and tissue used as controls is considered a necessary component in surgical pathology articles in which immunohistochemistry is utilized, such documentation seems less stringent in the cytopathologic literature. A comprehensive literature search was done for articles published in English within the last 15 years on nine of the most widely used antibodies in cytopathology. Individual case reports were excluded. Of the 100 articles reviewed, 13 articles were review articles or commentaries and hence not included in the analysis. Only 11 (13%) of the remaining 87 articles described positive and negative controls run on identically prepared samples. Forty-seven articles (54%) either did not mention controls or did not run controls as separate specimens. Sixteen articles (18%) included a vague statement about controls. Twelve (14%) commented only on the negative control, included only histology tissue controls, or included cell block controls, but the study also included other types of preparations, such as cytospins. One article (1%) did not include controls because of insufficient material. The College of American Pathologists recognizes the impracticality of maintaining separate positive control samples for every possible combination of fixation, processing, and specimen type. However, more stringent documentation of procedure and use of controls in the cytopathologic literature will ensure that immunocytochemistry results in diagnostic cytopathology as well as in research are valid and reproducible.

  4. 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.

  5. 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.

  6. Monoclonal antibodies against human BAP31 for immunocytochemistry.

    PubMed

    Song, Chaojun; Wang, Fuli; Xu, Zhuwei; Hu, Jintao; Tao, Haiqiang; Yang, Angang; Yang, Kun; Jin, Boquan

    2009-06-01

    Human BAP31 is a 28 kDa polytopic integral protein of the ER and part of a large BAP hetero-oligomeric complex that includes the related BAP29 protein and connections to actomyosin. BAP31 interacts with mIgD, cellubrevin, major histocompatibility complex class I, and BCL-2/BCL-X(L), and plays an important role in regulating the egress of these proteins and in apoptosis. Northern blot analyses have revealed BAP31 RNA transcripts in many tissues, including thymus, spleen, brain, kidney, testis, liver, and lung. However, prominent BAP31 protein expression analyzed by immunohistochemistry is restricted to a minority of cells in normal human tissue. Further studies should be made to verify the expression profiles of BAP31 in the protein level. Production of high affinity MAbs suitable for immunohistochemical staining has lagged. Here we generate a set of MAbs that could be used in Western blot, immunoprecipitation, and immunocytochemistry, providing a new powerful tool for investigation of expression profile of BAP31 protein and furthers the study of BAP31 functions.

  7. 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.

  8. Use of antibody fragments (Fv) in immunocytochemistry.

    PubMed

    Kleymann, G; Ostermeier, C; Heitmann, K; Haase, W; Michel, H

    1995-06-01

    We developed a novel antibody fragment (Fv) technique for localization and determination of the surface topology of membrane protein complexes by immunogold electron microscopy. Several hybridoma cell lines producing murine monoclonal antibodies (MAbs) raised against bacterial membrane proteins were established. The cDNAs coding for the variable domains of the MAbs were cloned and expressed in Escherichia coli. The engineered Fv fragments served as trifunctional adapter molecules. The Fv fragment binds to the epitope of the membrane protein. The Strep tag fused to the VH chain was used for one-step affinity purification of the Fv fragments. Immunological detection of the membrane protein-bound Fv fragments in electron microscopy was accomplished either via the Strep tag with colloidal gold-labeled streptavidin or via the c-myc tag, which was fused to the VL chain, in combination with the c-myc tag-specific antibody 9E10 and a colloidal gold-labeled secondary antibody. We examined four Fv fragments directed against the cytochrome c oxidase or the ubiquinol-cytochrome c oxidoreductase of Paracoccus denitrificans and bacteriorhodopsin of Halobacterium halobium to show that this method is generally applicable. In all cases the Fv fragments showed the same results as their corresponding parent antibodies in electron microscopic immunostaining and other applications.

  9. Medium temperature epoxy resin for immunocytochemistry: Quetol 651 with water.

    PubMed

    Abad, A R

    1992-02-01

    The addition of 1% water to the epoxy resin Quetol increased the labeling intensity of the sample. The significant decrease of the curing temperature of the epoxy resin may assist in preservation of antigens. Water may also reduce the cross-linkage of the resin allowing more antigen to be available to the antibodies. The modified Quetol resin is an option for use in immunocytochemistry studies.

  10. 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.

  11. Immunocytochemistry of the AfaE adhesin and AfaD invasin produced by pathogenic Escherichia coli strains during interaction of the bacteria with HeLa cells by high-resolution scanning electron microscopy.

    PubMed

    Gounon, P; Jouve, M; Le Bouguénec, C

    2000-04-01

    We used a recent scanning electron microscope equipped with field emission gun and highly sensitive detectors to develop a fast and simple protocol for double immunogold staining using 10- and 15-nm gold particles. We used this approach to analyse the afimbrial adhesive sheath produced by pathogenic Escherichia coli interacting with the surface of epithelial cells. We demonstrated that AfaE adhesin and AfaD invasin were exposed at the bacterial surface during the interaction. This method could be easily and widely extended to the study of the early invasion process of many bacterial and viral pathogens, by immunocytochemical probing.

  12. Differential diagnosis between mesothelioma and adenocarcinoma: a multimodal approach based on ultrastructure and immunocytochemistry

    SciTech Connect

    Bedrossian, C.W.; Bonsib, S.; Moran, C. )

    1992-05-01

    Most compensations for asbestos-related deaths secondary to cancer center around mesothelioma and bronchogenic carcinoma. The differential diagnosis between mesothelioma and adenocarcinoma is a common and troublesome one, necessitating the correlation between clinical history, radiographic findings, and pathologic examination of tissues and cells. We describe a multimodal approach based on the use of routine and special stains, immunocytochemistry, and electron microscopy for distinguishing between mesothelioma and adenocarcinoma. Once a malignant diagnosis is arrived at by careful pathological examination, the tumor is classified as mesothelioma if mesothelial cells are identified as the constituent cells of the neoplasm. Mesothelial cells are recognized by (1) their main ultrastructural features: slender and elongated microvilli, abundant intermediate filaments, and lacking secretory granules; and (2) their characteristic immunocytochemical reactivity: positivity for cytokeratin, EMA, and vimentin, and negativity for carcinoembryonic antigen (CEA), B72-3, Leu-M1, and other gland-cell markers. A variety of methods have been attempted in an effort to distinguish between reactive and malignant mesothelial cells. In practice, however, such distinction depends more on experience and expertise than in any fool-proof ancillary tests. A number of these tests are discussed along with the illustration of classical and unusual examples of mesothelioma and other pleural tumors.

  13. 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.

  14. [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.

  15. The naris muscles in tiger salamander. II. Innervation as revealed by enzyme histochemistry and immunocytochemistry.

    PubMed

    Wirsig-Wiechmann, Celeste R; Ebadifar, Bahareh

    2002-06-01

    The naris muscles control the aperature of the external naris in tiger salamanders, Ambystoma tigrinum, and may contribute to glandular secretion. Autonomic neurons of the palatine ganglion and possibly neurons associated with the nervus terminalis innervate these muscles. To elucidate the neural control of the naris muscles, neurotransmitters in nerve fibers supplying the naris muscles and in neurons of the palatine ganglion were examined using acetylcholinesterase enzyme histochemistry and immunocytochemistry to visualize possible peptide candidates for muscle innervation. The naris muscles, autonomic neurons, and associated nerve fascicles demonstrated strong acetylcholinesterase labeling, and the muscles were innervated by substance P fibers passing through the palatine ganglion from the trigeminal ganglion. Gonadotropin-releasing hormone and molluscan cardioexcitatory peptide-like immunoreactivities were found in secretory cell bodies and/or fibers in the palatine ganglion, and gonadotropin-releasing hormone was found in fiber projection pathways into the muscles. Vasoactive intestinal peptide was found in cell bodies and fibers of the palatine ganglion but appeared to provide a sparse innervation to the naris dilator muscle only. These findings suggest a typical autonomic cholinergic and sensory innervation of the naris muscles with some variations in peptide innervation. The presence of gonadotropin-releasing hormone in palatine ganglion and naris constrictor muscle suggests a potential modulation of autonomic neurons and perhaps even muscle fibers by this neuropeptide. We hypothesize that this reproductive hormone may modulate the activity of the naris constrictor muscle during reproductively appropriate events in order to provide access of pheromones to the vomeronasal organ.

  16. 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.

  17. The market for utility-scale fuel cell plants

    NASA Astrophysics Data System (ADS)

    Watanabe, Yasuo; Matsumoto, Masaru; Takasu, Kazuhiko

    This paper is devoted to a survey of the current technology and future market for utility-scale fuel cell plants. The phosphoric acid fuel cell (PAFC) is entering into the stage where it is practically available for use with natural gas. Large capacity plants such as 11, 5 and 1 MW have been installed and operated in Italy and Japan. Their efficiency ranges from 36 to 42%. The molten carbonate fuel cell (MCFC) is in the demonstrating stage, both the fuel cell and the balance-of-plant (BOP) for natural gas. Demonstration plants of 2 and 1 MW have been under construction in the USA and Japan. Their efficiency will range from 40 to 50%. The solid oxide fuel cell (SOFC) is in the experimental stage around 100 kW for co-generation. Its conceptual system design has been conducted for both centralized and dispersed power plant in a cooperation with Westinghouse and NEDO. A market survey is now considered on the basis that future fuel cells will run for around 40 000 h in a stable manner with competitive performance. The market for fuel cells will be roughly at 2000 MW in Japan by the year 2010. Half of them will be installed for electric companies on the utility scale. The market will be shared between PAFC and MCFC by 10 and 90%, respectively. Current technologies have not reached the stage to precisely forecast when fuel cells will be entering into the market on a utility scale. At the present time, it is worthwhile to consider how the technological and economic requirements will be definitely achieved. After achieving these requirements, fuel cells will be positively introduced and socially accepted as the best energy converting option to save energy and environmental impact. Further efforts will be devoted to meeting the market from the technological and economic aspects.

  18. 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.

  19. Keratin proteins in human lung carcinomas. Combined use of morphology, keratin immunocytochemistry, and keratin immunoprecipitation.

    PubMed Central

    Banks-Schlegel, S. P.; McDowell, E. M.; Wilson, T. S.; Trump, B. F.; Harris, C. C.

    1984-01-01

    Light-microscopic immunocytochemistry and electron microscopy demonstrated that adenocarcinomas (AC) and squamous cell (epidermoid) carcinomas (SCCs) of human lung contained keratin proteins in the form of tonofilament bundles. However, moderately differentiated (md) SCCs contained abundant keratin, whereas poorly differentiated (pd) SCCs and all ACs contained lesser amounts. Lung tumors with the diagnosis of AC or SCC, as defined by WHO criteria, were also analyzed by immunoprecipitation techniques for the presence of keratin proteins. Regardless of the degree of tumor differentiation, SCCs contained a 44 kd keratin which was lacking in ACs. Interestingly, normal bronchial epithelium also contained the same 44 kd keratin. In addition, as SCCs became more differentiated, they exhibited even greater differences in the profile of synthesized keratins. Specifically, the relative abundance of the intermediate-sized keratins (57 and 59 kd) was increased in the md SCCs. Although keratin protein patterns appear to be a valuable adjunct in distinguishing AC from SCC, their usefulness as a diagnostic tool will require survey of a larger number of poorly differentiated tumors. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 PMID:6198920

  20. 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.

  1. Utility of Transmission Electron Microscopy in Small Round Cell Tumors

    PubMed Central

    Kim, Na Rae; Ha, Seung Yeon; Cho, Hyun Yee

    2015-01-01

    Small round cell tumors (SRCTs) are a heterogeneous group of neoplasms composed of small, primitive, and undifferentiated cells sharing similar histology under light microscopy. SRCTs include Ewing sarcoma/peripheral neuroectodermal tumor family tumors, neuroblastoma, desmoplastic SRCT, rhabdomyosarcoma, poorly differentiated round cell synovial sarcoma, mesenchymal chondrosarcoma, small cell osteosarcoma, small cell malignant peripheral nerve sheath tumor, and small cell schwannoma. Non-Hodgkin’s malignant lymphoma, myeloid sarcoma, malignant melanoma, and gastrointestinal stromal tumor may also present as SRCT. The current shift towards immunohistochemistry and cytogenetic molecular techniques for SRCT may be inappropriate because of antigenic overlapping or inconclusive molecular results due to the lack of differentiation of primitive cells and unavailable genetic service or limited moleculocytogenetic experience. Although usage has declined, electron microscopy (EM) remains very useful and shows salient features for the diagnosis of SRCTs. Although EM is not always required, it provides reliability and validity in the diagnosis of SRCT. Here, the ultrastructural characteristics of SRCTs are reviewed and we suggest that EM would be utilized as one of the reliable modalities for the diagnosis of undifferentiated and poorly differentiated SRCTs. PMID:25812730

  2. 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.

  3. Immunocytochemistry of the amphibian embryo--from overview to ultrastructure.

    PubMed

    Kurth, Thomas

    2003-06-01

    Amphibian embryos are standard research objects to study pattern formation and morphogenesis. Due to their external development and robust nature, experimental manipulations such as microinjections or transplantations can be easily performed. However, most immunocytochemical approaches addressing the specific localization of proteins are hampered by the fragility of the large and yolky embryonic cells which render high resolution staining difficult. Immunocytochemical data are therefore often restricted to either overall patterns in whole embryo preparations or to immunofluorescent localization with limited resolution on sections. High resolution or ultrastructural protein localization data are rare and can be achieved only with time consuming procedures. Here, a comparative study of immunocytochemical methods suitable for light and electron microscopy using different kinds of plastic resins is presented. Three main approaches are described: preembedding staining of whole embryos, postembedding staining of ultrathin sections and preembedding staining of vibratome sections. All the procedures are designed to study protein expression in early amphibian embryos en gros as well as en detail and the described techniques are suitable to combine two or three levels of resolution on the very same biological specimen. Examples are presented and advantages and disadvantages of the different protocols are discussed.

  4. 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.

  5. Magnetic activated cell sorting (MACS): utility in assisted reproduction.

    PubMed

    Makker, Kartikeya; Agarwal, Ashok; Sharma, Rakesh K

    2008-07-01

    Assisted reproductive techniques (ART) have now been extensively incorporated in the management of infertile couples. But even after rapid methodological and technological advances the success rates of these procedures have been below expectations. This has led to development of many sperm preparation protocols to obtain an ideal semen sample for artificial reproduction. Sperm apoptosis has been heavily linked to failures in reproductive techniques. One of the earliest changes shown by apoptotic spermatozoa is externalization of phosphatidyl serine. Magnetic activated cell sorting (MACS) is a novel sperm preparation technique that separates apoptotic and non-apoptotic spermatozoa based on the expression of phosphatidylserine. This has led to the incorporation of MACS as a sperm preparation technique. The review highlights the principle and mechanism of this novel technique and enumerates its advantages as a sperm preparation technique. Its utility in ART as an efficient tool for sperm recovery and its application in cryopreservation of semen samples is also explained.

  6. 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.

  7. 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

  8. 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

  9. Live cell quality control and utility of real-time cell electronic sensing for assay development.

    PubMed

    Kirstein, Shelli L; Atienza, Josephine M; Xi, Biao; Zhu, Jenny; Yu, Naichen; Wang, Xiaobo; Xu, Xiao; Abassi, Yama A

    2006-10-01

    In this paper we have explored the utility of the real-time cell electronic sensing (RTCES, ACEA Biosciences Inc., San Diego, CA) system for monitoring the quality of live cells in cell-based assays as well as for assay development. We have demonstrated that each cell type displays unique growth kinetic profiles that provide a quantitative account of cell behavior and can be used as a diagnostic tool for cellular quality control. The utility of the specific signature patterns was shown by demonstrating the significant differences in primary cell behavior depending on the supplier. In addition, the RT-CES system was able to differentiate cell behavior depending on the passage stage of the cells. The utility of the RT-CES system as an assay development tool was demonstrated in cytotoxicity assays. The RT-CES system not only provides information regarding the potency of cytotoxic compounds, but in addition relates potency to the rate of the response for each concentration of the compound tested, which is important for understanding the mechanism of compound action. Moreover, real-time display of cytotoxicity data by the RT-CES system allows for calculation of real-time 50% inhibitory concentration (IC50) values or determination of optimal IC(50) value. In summary, the RT-CES system provides high content and information-rich data that are beyond the scope of single-point assays.

  10. Decisive role of immunocytochemistry in aspiration cytology of chordoma of the clivus: a case report with review of the literature.

    PubMed

    Gherardi, G; Marveggio, C; Cola, C; Redaelli, G

    1994-05-01

    Immunocytochemistry (ICC) proved to be an essential adjunct in the fine-needle aspiration (FNA) cytological diagnosis of chordoma of the clivus in a 62-year-old woman. The cytological picture in routinely stained smears was not entirely diagnostic for chordoma due to the paucity of typical 'physalipherous' cells. To exclude other primary or metastatic neoplasms of the skull base possibly sharing the same cytological picture, additional direct smears were immunostained with antibodies specific for cytokeratin (CK), vimentin (VIM), S100 protein (S100P), carcinoembrionic antigen (CEA), epithelial membrane antigen (EMA), glial fibrillary acidic protein (GFAP), CD68 antigen (KP1) and with the 'panepithelial' antibodies B72.3 and Ber-EP4. Chordoma cells showed the following immunoprofile: CK+/VIM+/S100P+/CEA-/EMA+/GFAP-/B72.3-/Ber-EP4-/CD68+. The pattern of immunoreactivity for CK, S100P and CEA confirms previously reported data, while the B72.3-/Ber-EP4-/CD68+ staining profile represents a novel observation. The detection of a CK+/S100+/CEA-/B72.3-/Ber-EP4- immunocytological profile of chordoma cells in aspirates is a basic requirement to exclude pertinent diagnostic differentials, such as metastatic carcinoma, ependymoma and sarcoma, and permits a reliable pre-operative diagnosis of the tumour by aspiration cytology.

  11. The effect of the small amount of formaldehyde in the SurePath liquid when establishing protocols for immunocytochemistry

    PubMed Central

    Bjønness-Jacobsen, Ellen Cathrine; Eriksen, Anne Kristine Kallevåg; Hagen, Vidar Nyløkken; Østbye, Kirsten Margrethe; Wittersø, Anna; Pedersen, Mette Kristin; Sauer, Torill

    2016-01-01

    Background: SurePath® is an ethanol-based liquid fixative. In addition to ethanol, it also contains a small amount of formaldehyde (<0.2%). The aim of this study was to investigate the immunoreactivity of cells stored for different lengths of time in the SurePath liquid. Materials and Methods: Rest material from one malignant and three benign effusions were fixed in SurePath for 1–12 days. Cytospins were incubated with cytokeratin 7 antibody (AB) to evaluate the staining intensity of carcinoma cells and benign, reactive mesothelial cells. Protocols varied as to pretreatment and AB incubation time. Results: Reduced immunostaining intensity was seen within 5 days of storage in the SurePath liquid. It was restored when the pretreatment time was prolonged. Conclusions: The small amount of formaldehyde in the SurePath liquid seems to affect the immunoreactivity. Local immunocytochemistry protocols in the cytology laboratories should consider this when optimizing their procedures. Postfixation with formalin should be omitted. PMID:27994636

  12. Fuel Cells for Electric Utility and Transportation Applications

    SciTech Connect

    Srinivasan, S.

    1980-01-01

    This review encompasses the following topics: (1) historical, (2) types of fuel cells, (3) thermodynamic and electrode kinetic aspects of fuel cells, (4) overview of present status of fuel cell research and development, (5) electrocatalysis of fuel cell reactions, (6) fuel cell/battery hybrid vehicles, and (7) regenerative hydrogen-halogen fuel cells for energy storage. (WHK)

  13. 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

  14. Cryoembedding and sectioning of cochleas for immunocytochemistry and in situ hybridization.

    PubMed

    Whitlon, D S; Szakaly, R; Greiner, M A

    2001-02-01

    Current emphasis on biochemical and molecular aspects of cochlear anatomy underscores the necessity for high quality cryostat sections of the inner ear. The large volume of fluid space within the cochlea makes cryoembedding and sectioning of the organ more problematic than that of other, more homogeneous tissues. Our method for cryoembedding of cochleas for immunocytochemistry and in situ hybridization uses slow infiltration with increasing concentrations of sucrose followed by degassed embedding medium before final orientation and freezing. This method permits high quality cryosections to be cut which preserve overall structure and cellular resolution.

  15. 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.

  16. Uveal melanoma cells utilize a novel route for transendothelial migration.

    PubMed

    Onken, Michael D; Li, Jinmei; Cooper, John A

    2014-01-01

    Uveal melanoma arises in the eye, and it spreads to distant organs in almost half of patients, leading to a fatal outcome. To metastasize, uveal melanoma cells must transmigrate into and out of the microvasculature, crossing the monolayer of endothelial cells that separates the vessel lumen from surrounding tissues. We investigated how human uveal melanoma cells cross the endothelial cell monolayer, using a cultured cell system with primary human endothelial cell monolayers on hydrogel substrates. We found that uveal melanoma cells transmigrate by a novel and unexpected mechanism. Uveal melanoma cells intercalate into the endothelial cell monolayer and flatten out, assuming a shape and geometry similar to those of endothelial cells in the monolayer. After an extended period of time in the intercalated state, the uveal melanoma cells round up and migrate underneath the monolayer. VCAM is present on endothelial cells, and anti-VCAM antibodies slowed the process of intercalation. Depletion of BAP1, a known suppressor of metastasis in patients, increased the amount of transmigration of uveal melanoma cells in transwell assays; but BAP1 depletion did not affect the rate of intercalation, based on movies of living cells. Our results reveal a novel route of transendothelial migration for uveal melanoma cells, and they provide insight into the mechanism by which loss of BAP1 promotes metastasis.

  17. Label-free Optofluidic Cell Classifier Utilizing Support Vector Machines.

    PubMed

    Wu, Tsung-Feng; Mei, Zhe; Lo, Yu-Hwa

    2013-09-01

    A unique optofluidic lab-on-a-chip device that can measure optically encoded forward scattering signals has been demonstrated. From the design of the spatial pattern, the position and velocity of each cell in the flow can be detected and then a spatial cell distribution over the cross section of the channel can be generated. According to the forward scattering intensity and position information of cells, a data-mining method, support vector machines (SVMs), is applied for cell classification. With the help of SVMs, the multi-dimensional analysis can be performed to significantly increase all figures of merit for cell classification.

  18. Downregulation of transgelin blocks interleukin-8 utilization and suppresses vasculogenic mimicry in breast cancer cells.

    PubMed

    Aikins, Anastasia R; Kim, MiJung; Raymundo, Bernardo; Kim, Chan-Wha

    2017-03-01

    Vasculogenic mimicry (VM) is a non-classical mechanism recently described in many tumors, whereby cancer cells, rather than endothelial cells, form blood vessels. Transgelin is an actin-binding protein that has been implicated in multiple stages of cancer development. In this study, we investigated the role of transgelin in VM and assessed its effect on the expression of endothelial and angiogenesis-related genes during VM in MDA-MB-231 breast cancer cells. We confirmed the ability of MDA-MB-231 cells to undergo VM through a tube formation assay. Flow cytometry analysis revealed an increase in the expression of the endothelial-related markers VE-cadherin and CD34 in cells that underwent VM, compared with those growing in a monolayer, which was confirmed by immunocytochemistry. We employed siRNA to silence transgelin, and knockdown efficiency was determined by western blot analyses. Downregulation of transgelin suppressed cell proliferation and tube formation, but increased IL-8 levels in Matrigel cultures. RT-PCR analyses revealed that the expression of IL-8, VE-cadherin, and CD34 was unaffected by transgelin knockdown, indicating that increased IL-8 expression was not due to enhanced transcriptional activity. More importantly, the inhibition of IL-8/CXCR2 signaling also resulted in suppression of VM with increased IL-8 levels, confirming that increased IL-8 levels after transgelin knockdown was due to inhibition of IL-8 uptake. Our findings indicate that transgelin regulates VM by enhancing IL uptake. These observations are relevant to the future development of efficient antivascular agents. Impact statement Vasculogenic mimicry (VM) is an angiogenic-independent mechanism of blood vessel formation whereby aggressive tumor cells undergo formation of capillary-like structures. Thus, interventions aimed at angiogenesis might not target the entire tumor vasculature. A more holistic approach is therefore needed in the development of improved antivascular agents

  19. 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

  20. 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.

  1. 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.

  2. Utilization of oil in soil and water by microbial cells

    SciTech Connect

    Surzhko, L.F.; Yankevich, M.I.; Yakovlev, V.I.

    1995-05-01

    Pure cultures and culture associations actively utilizing aliphatic and aromatic fractions of oil of the Romashkinskoe oil field were selected. Biopreparations based on vermiculite-immobilized active oil-degrading strains were developed. The ability of the preparations to decompose oil in soil and water was shown. The preparation containing an association of yeast and the bacterium Acinetobacter sp. 725 proved most efficient. In field lysimetric experiments, the introduction of the preparations accelerated oil decomposition by up to 65-78% compared to the control. 6 refs., 2 figs., 4 tabs.

  3. Identification, Characterization, and Utilization of Adult Meniscal Progenitor Cells

    DTIC Science & Technology

    2014-09-01

    year old mouse menisci. MSPCs grow as colonies, express stem cell and meniscal gene signature markers found in adult human meniscus, and can be...be collected from parallel cultures for measurement of meniscus signature genes , stem cell markers as well as markers that identify bone, cartilage...in control media from both 8wk and 6month old meniscal explants. We then used real time PCR to analyze gene expression. 0   1   2   3

  4. 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

  5. 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

  6. [Biomass energy utilization in microbial fuel cells: potentials and challenges].

    PubMed

    Huang, Liping; Cheng, Shaoan

    2010-07-01

    Microbial fuel cells (MFCs) that can harvest biomass energy from organic wastes through microbial catalysis have garnered more and more attention within the past decade due to its potential benefits to ecological environment. In this article, the updated progress in MFCs is reviewed, with a focus on frontier technologies such as chamber configurations, feedstock varieties and the integration of MFCs with microbial electrolysis cells for hydrogen production. And on the other hand, the challenges like development of cost-effective electrode materials, improvement of biomass energy recovery and power output, design and optimization of commercial MFC devices are presented.

  7. 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.

  8. 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.

  9. 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.

  10. Identification, Characterization, and Utilization of Adult Meniscal Progenitor Cells

    DTIC Science & Technology

    2015-09-01

    key to derailing the connection between acute meniscal injury and post-traumatic knee OA. Here we combine mouse genetics with molecular and cell...effective indentation modulus when compared to controls. Specific Aim 3. Task 3: Production of meniscus tear. One week after injection of genetically ...unique platform to study synovial joint development and OA pathology due to its short lifespan, low cost of maintenance and availability for genetic

  11. Plasmonic nanocrystal solar cells utilizing strongly confined radiation.

    PubMed

    Kholmicheva, Natalia; Moroz, Pavel; Rijal, Upendra; Bastola, Ebin; Uprety, Prakash; Liyanage, Geethika; Razgoniaev, Anton; Ostrowski, Alexis D; Zamkov, Mikhail

    2014-12-23

    The ability of metal nanoparticles to concentrate light via the plasmon resonance represents a unique opportunity for funneling the solar energy in photovoltaic devices. The absorption enhancement in plasmonic solar cells is predicted to be particularly prominent when the size of metal features falls below 20 nm, causing the strong confinement of radiation modes. Unfortunately, the ultrashort lifetime of such near-field radiation makes harvesting the plasmon energy in small-diameter nanoparticles a challenging task. Here, we develop plasmonic solar cells that harness the near-field emission of 5 nm Au nanoparticles by transferring the plasmon energy to band gap transitions of PbS semiconductor nanocrystals. The interfaces of Au and PbS domains were designed to support a rapid energy transfer at rates that outpace the thermal dephasing of plasmon modes. We demonstrate that central to the device operation is the inorganic passivation of Au nanoparticles with a wide gap semiconductor, which reduces carrier scattering and simultaneously improves the stability of heat-prone plasmonic films. The contribution of the Au near-field emission toward the charge carrier generation was manifested through the observation of an enhanced short circuit current and improved power conversion efficiency of mixed (Au, PbS) solar cells, as measured relative to PbS-only devices.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. Analysis by immunocytochemistry and in situ hybridization of renin and its mRNA in kidney, testis, adrenal, and pituitary of the rat.

    PubMed Central

    Deschepper, C F; Mellon, S H; Cumin, F; Baxter, J D; Ganong, W F

    1986-01-01

    Renin gene expression in cells and tissues of the rat was examined by in situ hybridization histochemistry and immunocytochemistry. By using a mouse cDNA probe, hybridization histochemistry revealed renin mRNA in the renal juxtaglomerular cells, testicular Leydig cells, adrenal zona glomerulosa cells, the intermediate lobe of the pituitary, and scattered cells of the anterior lobe of the pituitary. With four separate antisera to mouse submaxillary renin, there was immunoreactivity in the renal juxtaglomerular cells. However, only one of the antisera stained the Leydig cells, a second stained the adrenal zona glomerulosa, a third stained the intermediate lobe of the pituitary, and a fourth stained scattered cells of the anterior lobe of the pituitary that were identified as gonadotrophs. The variations with the different antisera in detecting extrarenal renin are unexplained but could imply that posttranslational proteolysis or glycosylation of preprorenin varies in different tissues with consequent variations in immunoreactivity. The finding of renin mRNA and renin-like immunoreactivity in these tissues supports the notion that these tissues are sites for production of renin. Images PMID:3532116

  18. A balloon ozone measurement utilizing an optical absorption cell and an ejector air sampler

    NASA Technical Reports Server (NTRS)

    Hilsenrath, E.; Ashenfelter, T. E.

    1976-01-01

    Stratospheric ozone was measured from a balloon utilizing an ultraviolet absorption cell. The ambient air was sampled by means of an aspirator attached to the output end of the optical cell. A nominal ozone distribution was obtained from 16 km to the float altitude of 38 km.

  19. 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.

  20. Comparison of Immunocytochemistry and Immunohistochemistry on Breast Carcinoma: A Boon or a Bane?

    PubMed Central

    Geethamala, Kempula; Murthy, Venkataramappa Srinivasa; Vani, Bangalore Ramalingiah; Rao, Madireddi Sudha; Thejaswini, Malugnalli Uddappa; Padmaja, Krishnarajapet Padmanabha

    2017-01-01

    Introduction: Breast carcinoma is the most common cancer among women in the urban Indian population. Conventionally, immunohistochemistry (IHC) is done to determine the hormone receptor status of the tumor. Immunocytochemistry (ICC) on fine-needle aspiration cytology (FNAC) was carried out to determine the same hormone receptor status of the tumor. Objective: The study was undertaken to evaluate the diagnostic reliability of performing estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (Her2/neu) receptor status on FNAC by ICC and to compare the results with IHC. Materials and Methods: A 2 years 6 months prospective study conducted in the Department of Pathology, ESIC Medical College and PGIMSR and ESIC Model Hospital, Rajajinagar, Bengaluru, wherein 100 breast carcinoma patients' samples both cytology and histology were collected. IHC and ICC were done by peroxidase antiperoxidase technique. Validations of the receptor status were analyzed using sensitivity, specificity, positive and negative predictive values (PPV and NPV), and kappa statistics for agreements between ICC and IHC. Results: ICC was positive for ER, PR, and Her2/neu in 53, 50, and 22 cases, respectively. For ER, a cytohistologic correlation of 98%, with a sensitivity of 96.3%, specificity of 100%, and PPV and NPV being 100% and 95.7%. For PR, concordance of 97%, with a sensitivity of 94.3%, specificity of 100%, and PPV and NPV being 100% and 94%. Her2/neu had an agreement of 89%, with a sensitivity of 72%, specificity of 95.5%, and PPV and NPV being 85.7% and 90.1%. Conclusion: ICC has been a boon and can be a paramount diagnostic adjunct to the routine investigations. PMID:28042209

  1. The use, publication and future directions of immunocytochemistry in veterinary medicine: a consensus of the Oncology-Pathology Working Group.

    PubMed

    Priest, H L; Hume, K R; Killick, D; Kozicki, A; Rizzo, V L; Seelig, D; Snyder, L A; Springer, N L; Wright, Z M; Robat, C

    2016-03-22

    One of the primary objectives of the Oncology Pathology Working Group (OPWG), a joint initiative of the Veterinary Cancer Society and the American College of Veterinary Pathologists, is for oncologists and pathologists to collaboratively generate consensus documents to standardize aspects of and provide guidelines for oncologic pathology. Consensus is established through review of relevant peer-reviewed literature relative to a subgroup's particular focus. In this document, the authors provide descriptions of the literature reviewed, the review process, and a summary of the information gathered on immunocytochemistry. The intent of this publication is to help educate practitioners and pathologists on the process of immunocytochemistry and to provide a guide for the use of this technique in veterinary medicine. This document represents the opinions of the working group and the authors and does not constitute a formal endorsement by the American College of Veterinary Pathologists or the Veterinary Cancer Society.

  2. Uptake and utilization of CDP-choline in primary brain cell cultures from fetal brain.

    PubMed

    Vecchini, A; Binaglia, L; Floridi, A; Palmerini, C A; Procellati, G

    1983-03-01

    The utilization of double-labeled CDP-choline by cultured brain cells has been studied. CDP-choline is demonstrated to be rapidly hydrolysed into CMP and choline phosphate. The fragments, or their hydrolysis products, penetrate into the cells and are utilized for lipid synthesis. At short times after the isotope administration a rapid labeling of phosphatidylcholine was detected, when cells were incubated with CDP-choline. The same was not seen when cells were incubated with labeled choline. From these observations it can be inferred that either CDP- choline can penetrate the cell membrane or that some mechanism involving CDP-choline and leading to phospholipid synthesis can work at the external surface of the plasma membranes.

  3. 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.

  4. Organoids from adult liver and pancreas: Stem cell biology and biomedical utility.

    PubMed

    Hindley, Christopher J; Cordero-Espinoza, Lucía; Huch, Meritxell

    2016-12-15

    The liver and pancreas are critical organs maintaining whole body metabolism. Historically, the expansion of adult-derived cells from these organs in vitro has proven challenging and this in turn has hampered studies of liver and pancreas stem cell biology, as well as being a roadblock to disease modelling and cell replacement therapies for pathologies in these organs. Recently, defined culture conditions have been described which allow the in vitro culture and manipulation of adult-derived liver and pancreatic material. Here we review these systems and assess their physiological relevance, as well as their potential utility in biomedicine.

  5. 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

  6. 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.

  7. 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.

  8. 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

  9. Comparison of three cell fixation methods for high content analysis assays utilizing quantum dots.

    PubMed

    Williams, Y; Byrne, S; Bashir, M; Davies, A; Whelan, A; Gun'ko, Y; Kelleher, D; Volkov, Y

    2008-10-01

    Semiconductor nanoparticles or quantum dots are being increasingly utilized as fluorescent probes in cell biology both in live and fixed cell assays. Quantum dots possess an immense potential for use in multiplexing assays that can be run on high content screening analysers. Depending on the nature of the biological target under investigation, experiments are frequently required on cells retaining an intact cell membrane or also on those that have been fixed and permeabilized to expose intracellular antigens. Fixation of cell lines before or after the addition of quantum dots may affect their localization, emission properties and stability. Using a high content analysis platform we perform a quantitative comparative analysis of three common fixation techniques in two different cell lines exposed to carboxylic acid stabilized CdTe quantum dots. Our study demonstrates that in prefixed and permeabilized cells, quantum dots are readily internalized regardless of cell type, and their intracellular location is primarily determined by the properties of the quantum dots themselves. However, if the fixation procedures are preformed on live cells previously incubated with quantum dots, other important factors have to be considered. The choice of the fixative significantly influences the fluorescent characteristics of the quantum dots. Fixatives, regardless of their chemical nature, negatively affected quantum dots fluorescence intensity. Comparative analysis of gluteraldehyde, methanol and paraformaldehyde demonstrated that 2% paraformaldehyde was the fixative of choice. The presence of protein in the media did not significantly alter the quantum dot fluorescence. This study indicates that multiplexing assays utilizing quantum dots, despite being a cutting edge tool for high content cell imaging, still require careful consideration of the basic steps in biological sample processing.

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

    PubMed

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

    2015-04-01

    Autologous hematopoietic cell transplantation (AHCT) is an established therapy for multiple myeloma (MM), with an impact on quality of remission and survival. We analyzed the role of race, ethnicity, sex, and age disparities in AHCT utilization in the United States. We combined MM incidence derived from the Surveillance, Epidemiology and End Results program with transplantation activity reported to the Center for International Blood and Marrow Transplant Research for the period of 2005 to 2009 to assess the impact of disparities in AHCT. Utilization (number of transplantations/new cases) was compared between groups using the relative utilization ratio (RUR), defined as [utilization for a given category]/[utilization for the entire population]. Data were obtained from 22,462 actual MM cases and 13,311 AHCT. The age-adjusted RUR was 1.17 (95% confidence interval [CI], 1.15 to 1.19) among non-Hispanic Whites (NHW), higher than in non-Hispanic Blacks (NHB) (age-adjusted RUR, .69; 95% CI, .67 to .72; P < .0002), Hispanics (age-adjusted RUR, .64; 95% CI, .60 to .69; P < .002), and Asians (age-adjusted RUR, .65; 95% CI, .58 to .73; P < .0002]. AHCT utilization was higher in men than in women among Hispanics (age-adjusted RUR .72 versus .56, P = .007), but not among NHW, NHB, or Asians. Sex disparity prevents 1.3% of potential AHCTs in patients with MM (10.4% among Hispanics). Racial-ethnic disparities prevent 13.8% of AHCTs (44.7% in Hispanic and Asians, 39.9% in NHBs). 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. Two-year and 5-year increases in the age of the AHCT population would result in 12% and 32% increases, respectively, in volume of AHCT.

  11. 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.

  12. Rapid isolation and detection of cancer cells by utilizing integrated microfluidic systems.

    PubMed

    Lien, Kang-Yi; Chuang, Ying-Hsin; Hung, Lein-Yu; Hsu, Keng-Fu; Lai, Wu-Wei; Ho, Chung-Liang; Chou, Cheng-Yang; Lee, Gwo-Bin

    2010-11-07

    The present study reports a new three-dimensional (3D) microfluidic platform capable of rapid isolation and detection of cancer cells from a large sample volume (e.g. ~1 mL) by utilizing magnetic microbead-based technologies. Several modules, including a 3D microfluidic incubator for the magnetic beads to capture cancer cells, a microfluidic control module for sample transportation and a nucleic acid amplification module for genetic identification, are integrated into this microsystem. With the incorporation of surface-modified magnetic beads, target cancer cells can be specifically recognized and conjugated onto the surface of the antibody-coated magnetic microbeads by utilizing a swirling effect generated by the new 3D microfluidic incubator, followed by isolating and purifying the magnetic complexes via the incorporation of an external magnet and a microfluidic control module, which washes away any unbound waste solution. Experimental results show that over 90% of the target cancer cells can be isolated from a large volume of bio-samples within 10 min in the 3D microfluidic incubator. In addition, the expressed genes associated with ovarian and lung cancer cells can also be successfully amplified by using the on-chip nucleic acid amplification module. More importantly, the detection limit of the developed system is found to be 5 × 10(1) cells mL(-1) for the target cancer cells, indicating that this proposed microfluidic system may be adapted for clinical use for the early detection of cancer cells. Consequently, the proposed 3D microfluidic system incorporated with immunomagnetic beads may provide a promising automated platform for the rapid isolation and detection of cancer cells with a high sensitivity.

  13. Genetically Targeted Radiotherapy Utilizing the Human Sodium Iodide Symporter in Human Breast Carcinoma Cells

    DTIC Science & Technology

    2005-04-01

    to concentrate isotopes has been utilized clinically for years in thyroid imaging, treatment of hyperthyroidism , and for treating well-differentiated...expression after retinoic acid treatments . This treatment may turn on lactoperoxidase expression as well increasing the retention of radioactivity in the...tumor. LPO treatment did increase retention of 1-125 in Ad-NIS treated cells compared to Ad-NIS treatment alone. The acquisition of a pin-hole

  14. 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.

  15. 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.

  16. A circadian neuropeptide, pigment-dispersing factor-PDF, in the last-summer cicada Meimuna opalifera: cDNA cloning and immunocytochemistry.

    PubMed

    Sato, Seiji; Chuman, Yoshiro; Matsushima, Ayami; Tominaga, Yoshiya; Shimohigashi, Yasuyuki; Shimohigashi, Miki

    2002-08-01

    Pigment-dispersing factor (PDF), an 18-amino acid neuropeptide, is a principal circadian neuromodulator functioning downstream of the insect brain's circadian clock, modulating daily rhythms of locomotor activity. Recently, we found that PDF precursors of the cricket Gryllus bimaculatus comprise a nuclear localization signal (NLS). Moreover, the nuclear localization of PDF immunoreactivity and the translocation of GFP-fused PDF precursor into the nucleus have both been demonstrated. These suggest a fundamental role for PDF peptide in the circadian clock system within the nucleus, in addition to its role in downstream neural events. In the present study, we carried out the cDNA cloning of PDF from adult brains of the last-summer cicada Meimuna opalifera, and found that an isolated clone (545 bp) encodes an ordinary PDF precursor protein. PDF peptide itself shows a high sequence identity (78-94%) and similarity (89-100%) to insect PDFs and also to the crustacean beta-PDH peptides. The computer-assisted sequence analysis of PDF precursor revealed a possible translocation into the nucleus, despite the lack of a definite NLS-like sequence. Using immunocytochemistry, the optic lobes of M. opalifera revealed PDF-immunoreactive neurons in both the medulla and lamina neuropiles. All these PDF cells exhibited prominent immunolabeling of both their perikarya and axons, but not their nuclei. Our results provide the first structural and immunocytochemical identification of PDF neurons in Hemiptera.

  17. Immunocytochemistry of endothelial nitric oxide synthase in the rat brain: a light and electron microscopical study using the tyramide signal amplification technique.

    PubMed

    Stanarius, A; Töpel, I; Schulz, S; Noack, H; Wolf, G

    1997-11-01

    There are many inconsistencies in the literature about the cellular and subcellular distribution of the endothelial isoform of nitric oxide synthase (eNOS) in the brain. We have re-investigated its localization by light and electron microscopical (LM, EM) immunocytochemistry and the NADPH-diaphorase reaction. Using bovine aortic tissue as a positive control the protocols for the fixation and staining procedure were optimized. Only cryosections immersion-fixed with aceton and a mixture of aldehydes exhibited a clear-cut immunostaining. In rat brain tissue the endothelium of the entire vasculature showed immunoreactivity and, in addition to that, the epithelial cells of the choroid plexuses, whereas neurons never displayed any signs of immunostaining. EM immunoprecipitates were seen irregularly distributed in the cytosol or attached to endocellular membranes. EM NADPH-diaphorase histochemistry using the tetrazolium salt BSPT provided incoherent pictures in so far as the reaction product was exclusively bound to membranes. The restriction of eNOS within brain tissue to the vasculature may have implications for the differential significance of NOS isoforms in brain function.

  18. Physiological features of periodontal regeneration and approaches for periodontal tissue engineering utilizing periodontal ligament cells.

    PubMed

    Benatti, Bruno Braga; Silvério, Karina Gonzales; Casati, Márcio Zaffalon; Sallum, Enílson Antônio; Nociti, Francisco Humberto

    2007-01-01

    Experimental studies have shown that the potential of periodontal regeneration seems to be limited by the regenerative capacity of the cells involved. The regeneration of damaged periodontal tissues is mediated by various periodontal cells and is regulated by a vast array of extracellular matrix informational molecules that induce both selective and nonselective responses in different cell lineages and their precursors. In this paper, we first review periodontal ligament tissue and its different cell subpopulations including fibroblasts and paravascular stem cells, and their functions during the development and homeostasis of periodontal tissues. Because conventional periodontal regeneration methods remain insufficient to obtain a complete and reliable periodontal regeneration, the concept of periodontal tissue engineering has been based on the generation of the conditions necessary to improve the healing of periodontal tissues. Additionally, the potential of periodontal ligament cells for use in periodontal tissue engineering to overcome the limitations of conventional periodontal regenerative therapies is discussed, followed by an update of the recent progress and future directions of research utilizing periodontal ligament cells for predictable periodontal regeneration.

  19. Utilization of ascites plasma very low density lipoprotein triglycerides by Ehrlich cells.

    PubMed

    Brenneman, D E; Spector, A A

    1974-07-01

    Much of the lipid present in the ascites plasma in which Ehrlich cells grow is contained in very low density lipoproteins (VLDL). Chemical measurements indicated that triglycerides were taken up by the cells during in vitro incubation with ascites VLDL. When tracer amounts of radioactive triolein were incorporated into the ascites VLDL, the percentage uptakes of glyceryl tri[1-(14)C]oleate and triglycerides measured chemically were similar. The cells also took up [2-(3)H]glyceryl trioleate that was added to VLDL, but the percentage of available (3)H recovered in the cell lipids was 30-40% less than that of (1 4)C from glyceryl tri[1-(1 4)C]oleate. This difference was accounted for by water-soluble (3)H that accumulated in the incubation medium, suggesting that extensive hydrolysis accompanied the uptake of VLDL triglycerides. Radioactive fatty acids derived from the VLDL triglycerides were incorporated into cell phospholipids, glycerides, and free fatty acids, and they also were oxidized to CO(2). Triglyceride utilization increased as the VLDL concentration was raised. These results suggest that one function of the ascites plasma VLDL may be to supply fatty acid to the Ehrlich cells and that the availability of fatty acid to this tumor is determined in part by the ascites plasma VLDL concentration. Although Ehrlich cells incorporate almost no free glycerol into triglycerides, considerable amounts of [2-(3)H]glyceryl trioleate radioactivity were recovered in cell triglycerides. This indicates that at least some VLDL triglycerides were taken up intact. The net uptake of VLDL protein and cholesterol was very small relative to the triglyceride uptake, suggesting that intact triglycerides are transferred from the ascites VLDL to the Ehrlich cells and that hydrolysis occurs after the triglyceride is associated with the cells.

  20. Pancreatic (acinar) metaplasia of the gastric mucosa. Histology, ultrastructure, immunocytochemistry, and clinicopathologic correlations of 101 cases.

    PubMed

    Doglioni, C; Laurino, L; Dei Tos, A P; De Boni, M; Franzin, G; Braidotti, P; Viale, G

    1993-11-01

    The occasional finding within the gastric mucosa of unidentified epithelial cells with morphological features closely resembling those of pancreatic acinar cells has prompted us to investigate a retrospective series of 8,430 consecutive gastric biopsies and of 126 surgical specimens of gastric resection and total gastrectomy. The aims of the study were to morphologically and immunocytochemically characterize these cells, to define their actual prevalence in a large series of unselected cases, and to assess the clinicopathologic correlates of their occurrence. Pancreatic acinar-like cells characterized by abundant cytoplasm, which was acidophilic and finely granular in the apical and middle portions and basophilic in the basal compartment, have been identified in 101 cases (84 gastric biopsies and 17 gastrectomies). These cells, arranged in nests or in variably sized lobules among the gastric glands, were morphologically indistinguishable from pancreatic acinar cells, both by light and by electron microscopy. Furthermore, they were consistently immunoreactive for pancreatic lipase and trypsinogen and, in 75% of the cases, for pancreatic alpha-amylase. The appearance of these cells within the gastric mucosa was correlated significantly with chronic gastritis (p = 0.032) and with the simultaneous occurrence of intestinal and pyloric types of gastric metaplasia (p = 0.021). The findings indicate that this is a previously unrecognized pancreatic (acinar) metaplasia of the gastric mucosa, clinically and morphologically distinct from pancreatic heterotopia.

  1. 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.

  2. 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.

  3. 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.

  4. Immunotherapy for Lewis lung carcinoma utilizing dendritic cells infected with CK19 gene recombinant adenoviral vectors

    PubMed Central

    SUN, Q.F.; ZHAO, X.N.; PENG, C.L.; HAO, Y.T.; ZHAO, Y.P.; JIANG, N.; XUE, H.; GUO, J.Z.; YUN, C.H.; CONG, B.; ZHAO, X.G.

    2015-01-01

    Dendritic cells (DCs) as 'professional' antigen-presenting cells (APCs) initiate and regulate immune responses to various antigens. DC-based vaccines have become a promising modality in cancer immunotherapy. Cytokeratin 19 (CK19) protein is expressed at high levels in lung cancer and many other tumor cells, suggesting CK19 as a potential tumor-specific target for cancer immune therapy. We constructed a recombinant adenoviral vector containing the CK19 gene (rAd-CK19). DCs transfected with rAd-CK19 were used to vaccinate C57BL/6 mice bearing xenografts derived from Lewis lung carcinoma (LLC) cells. The transfected DCs gave rise to potent CK19-specific cytotoxic T lymphocytes (CTLs) capable of lysing LLC cells. Mice immunized with the rAd-CK19-DCs exhibited significantly attenuated tumor growth (including tumor volume and weight) when compared to the tumor growth of mice immunized with rAd-c DCs or DCs during the 24-day observation period (P<0.05). The results revealed that the mice vaccinated with the rAd-CK19-DCs exhibited a potent protective and therapeutic antitumor immunity to LLC cells in the subcutaneous model along with an inhibitive effect on tumor growth compared to the mice vaccinated with the rAd-c DCs or DCs alone. The present study proposes a meaningful mode of action utilizing rAd-CK19 DCs in lung cancer immunotherapy. PMID:26323510

  5. Oligodendrocyte Progenitor Cells Directly Utilize Lactate for Promoting Cell Cycling and Differentiation

    PubMed Central

    Ichihara, Yoshinori; Doi, Toru; Ryu, Youngjae; Nagao, Motoshi; Sawada, Yasuhiro

    2016-01-01

    Oligodendrocyte progenitor cells (OPCs) undergo marked morphological changes to become mature oligodendrocytes, but the metabolic resources for this process have not been fully elucidated. Although lactate, a metabolic derivative of glycogen, has been reported to be consumed in oligodendrocytes as a metabolite, and to ameliorate hypomyelination induced by low glucose conditions, it is not clear about the direct contribution of lactate to cell cycling and differentiation of OPCs, and the source of lactate for remyelination. Therefore, we evaluated the effect of 1,4‐dideoxy‐1,4‐imino‐d‐arabinitol (DAB), an inhibitor of the glycogen catabolic enzyme glycogen phosphorylase, in a mouse cuprizone model. Cuprizone induced demyelination in the corpus callosum and remyelination occurred after cuprizone treatment ceased. This remyelination was inhibited by the administration of DAB. To further examine whether lactate affects proliferation or differentiation of OPCs, we cultured mouse primary OPC‐rich cells and analyzed the effect of lactate. Lactate rescued the slowed cell cycling induced by 0.4 mM glucose, as assessed by the BrdU‐positive cell ratio. Lactate also promoted OPC differentiation detected by monitoring the mature oligodendrocyte marker myelin basic protein, in the presence of both 36.6 mM and 0.4 mM glucose. Furthermore, these lactate‐mediated effects were suppressed by the reported monocarboxylate transporter inhibitor, α‐cyano‐4‐hydroxy‐cinnamate. These results suggest that lactate directly promotes the cell cycling rate and differentiation of OPCs, and that glycogen, one of the sources of lactate, contributes to remyelination in vivo. J. Cell. Physiol. 232: 986–995, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. PMID:27861886

  6. Oligodendrocyte Progenitor Cells Directly Utilize Lactate for Promoting Cell Cycling and Differentiation.

    PubMed

    Ichihara, Yoshinori; Doi, Toru; Ryu, Youngjae; Nagao, Motoshi; Sawada, Yasuhiro; Ogata, Toru

    2017-05-01

    Oligodendrocyte progenitor cells (OPCs) undergo marked morphological changes to become mature oligodendrocytes, but the metabolic resources for this process have not been fully elucidated. Although lactate, a metabolic derivative of glycogen, has been reported to be consumed in oligodendrocytes as a metabolite, and to ameliorate hypomyelination induced by low glucose conditions, it is not clear about the direct contribution of lactate to cell cycling and differentiation of OPCs, and the source of lactate for remyelination. Therefore, we evaluated the effect of 1,4-dideoxy-1,4-imino-d-arabinitol (DAB), an inhibitor of the glycogen catabolic enzyme glycogen phosphorylase, in a mouse cuprizone model. Cuprizone induced demyelination in the corpus callosum and remyelination occurred after cuprizone treatment ceased. This remyelination was inhibited by the administration of DAB. To further examine whether lactate affects proliferation or differentiation of OPCs, we cultured mouse primary OPC-rich cells and analyzed the effect of lactate. Lactate rescued the slowed cell cycling induced by 0.4 mM glucose, as assessed by the BrdU-positive cell ratio. Lactate also promoted OPC differentiation detected by monitoring the mature oligodendrocyte marker myelin basic protein, in the presence of both 36.6 mM and 0.4 mM glucose. Furthermore, these lactate-mediated effects were suppressed by the reported monocarboxylate transporter inhibitor, α-cyano-4-hydroxy-cinnamate. These results suggest that lactate directly promotes the cell cycling rate and differentiation of OPCs, and that glycogen, one of the sources of lactate, contributes to remyelination in vivo. J. Cell. Physiol. 232: 986-995, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc.

  7. Delta9-tetrahydrocannabinol stimulates glucose utilization in C6 glioma cells.

    PubMed

    Sánchez, C; Velasco, G; Guzmán, M

    1997-08-29

    The present work was undertaken to study the metabolic response of C6 glioma cells to physiologically relevant doses of delta9-tetrahydrocannabinol (THC), the major active component of marijuana. At those concentrations (i.e. nanomolar range), THC produced a dose-dependent increase in the rates of glucose oxidation to CO2 and glucose incorporation into phospholipids and glycogen. The THC-induced stimulation of glucose utilization was (i) dose-dependent up to 100 nM THC, (ii) mimicked by the synthetic cannabinoid HU-210, and (iii) prevented by pertussis toxin and the CB1 receptor antagonist SR141716A. In contrast to THC, forskolin markedly depressed CO2 production, phospholipid synthesis and glycogen synthesis from glucose. The forskolin-induced inhibition of glucose utilization was (i) mimicked by dibutyryl-cAMP, and (ii) prevented by THC, HU-210 and H-7, an inhibitor of the cAMP-dependent protein kinase. Likewise, THC was able to antagonize in part the forskolin-induced elevation of intracellular cAMP concentration, and this antagonistic effect was prevented by SR141716A. However, THC per se did not affect basal cAMP concentration. Results thus indicate that physiologically relevant doses of THC stimulate glucose metabolism in C6 glioma cells through a cannabinoid receptor-mediated process. Although cannabinoid receptors may be coupled to inhibition of adenylyl cyclase in C6 glioma cells, this does not seem to be the mechanism involved in the THC-induced stimulation of glucose metabolism.

  8. An osmium-free method of epon embedment that preserves both ultrastructure and antigenicity for post-embedding immunocytochemistry.

    PubMed

    Phend, K D; Rustioni, A; Weinberg, R J

    1995-03-01

    Immunocytochemistry for amino acids with post-embedding gold is compatible with glutaraldehyde fixation, osmication, and embedding in epoxy-based plastics, but immunogold detection of larger molecules in the central nervous system commonly requires special procedures, e.g. minimizing exposure to glutaraldehyde, eliminating osmium, cryosectioning, and/or embedding in acrylic plastics. These make samples more difficult to prepare and view and may compromise structural preservation. We report a new technique, fixing with high levels of glutaraldehyde, replacing osmium with tannic acid followed by other heavy metals and p-phenylenediamine, and embedding in Epon. This method optimizes antigenicity while retaining the structural preservation and convenient handling of standard embedding techniques. Compared to standard Epon embedment, labeling for neuropeptides in brain and spinal cord is improved. Moreover, the present method yields excellent labeling of glutamate receptors (difficult to identify with traditional post-embedding techniques) and enables simultaneous visualization of associated neurotransmitters.

  9. 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

  10. Bioelectrocatalysts: engineered oxidoreductase system for utilization of fumarate reductase in chemical synthesis, detection, and fuel cells.

    PubMed

    Park, Doo Hyun; Vieille, C; Zeikus, J G

    2003-10-01

    Fumarate reductase was used as a model oxidoreductase to demonstrate continuous electrical cofactor reduction-oxidation during the bioelectrochemical synthesis and detection of chemicals. The enzyme preparation was immobilized onto a graphite felt electrode that was modified with carboxymethylcellulose (CMC). Nicotinamide adenine dinucleotide (NAD), neutral red, and fumarate reductase (which contained menaquinone) were covalently linked by peptide bonds to the CMC. The electron mediator neutral red allowed NAD and menaquinone to be recycled electrically during enzymatic chemical synthesis. Succinate detection by the bioelectrocatalyst was linear from 5 microM to 10 mM succinate. Fumarate synthesis using this bioelectrode was dependent on succinate utilization and resulted in proportional production of electricity and fumarate. Succinate synthesis using this bioelectrocatalyst was dependent on current and fumarate concentration. This bioelectrocatalyst system may enhance the utility of menaquinone- and/or pyridine nucleotide-linked oxidoreductases in diverse enzymatic fuel cells and sensors. It may also enhance the utility of oxidoreductase-based chemical synthesis systems because it eliminates the problem of cofactor recycling.

  11. 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.

  12. 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.

  13. 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

  14. Pig models of neurodegenerative disorders: Utilization in cell replacement-based preclinical safety and efficacy studies.

    PubMed

    Dolezalova, Dasa; Hruska-Plochan, Marian; Bjarkam, Carsten R; Sørensen, Jens Christian H; Cunningham, Miles; Weingarten, David; Ciacci, Joseph D; Juhas, Stefan; Juhasova, Jana; Motlik, Jan; Hefferan, Michael P; Hazel, Tom; Johe, Karl; Carromeu, Cassiano; Muotri, Alysson; Bui, Jack; Strnadel, Jan; Marsala, Martin

    2014-08-15

    An important component for successful translation of cell replacement-based therapies into clinical practice is the utilization of large animal models to conduct efficacy and/or safety cell dosing studies. Over the past few decades, several large animal models (dog, cat, nonhuman primate) were developed and employed in cell replacement studies; however, none of these models appears to provide a readily available platform to conduct effective and large-scale preclinical studies. In recent years, numerous pig models of neurodegenerative disorders were developed using both a transgenic approach as well as invasive surgical techniques. The pig model (naïve noninjured animals) was recently used successfully to define the safety and optimal dosing of human spinal stem cells after grafting into the central nervous system (CNS) in immunosuppressed animals. The data from these studies were used in the design of a human clinical protocol used in amyotrophic lateral sclerosis (ALS) patients in a Phase I clinical trial. In addition, a highly inbred (complete major histocompatibility complex [MHC] match) strain of miniature pigs is available which permits the design of comparable MHC combinations between the donor cells and the graft recipient as used in human patients. Jointly, these studies show that the pig model can represent an effective large animal model to be used in preclinical cell replacement modeling. This review summarizes the available pig models of neurodegenerative disorders and the use of some of these models in cell replacement studies. The challenges and potential future directions in more effective use of the pig neurodegenerative models are also discussed.

  15. Immunocytochemistry of band 3 protein in kidney and other tissues of control and cystic fibrosis patients.

    PubMed

    Hazen-Martin, D J; Pasternack, G; Hennigar, R A; Spicer, S S; Sens, D A

    1987-03-01

    Current evidence indicates that the underlying genetic defect in cystic fibrosis (CF) results in defective chloride transport, and more specifically, chloride impermeability. However, recent consideration has been given to a possible defect in band 3 protein at CF-affected sites. In an effort to determine a possible role for band 3, an anion exchange channel protein, a series of immunocytochemical localizations was performed. Immunocytochemical staining for the anion channel band 3, 43K, in normal and CF human kidney was confined to erythrocytes, glomerular podocytes, the basal region of half of the cells of the initial connecting segment of cortical collecting ducts, and a minority of cells in medullary ray-collecting ducts. Erythrocytes alone evidenced immunoreactivity for band 3 protein in human pancreas, submandibular gland, trachea, and lung. In all cases, specimens from patients with CF stained like those from control subjects for band 3. Abnormal Cl- transport in CF tissues is not reflected in altered band 3, 43K, localizations.

  16. 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

  17. Circulating Tumor Cells, DNA, and mRNA: Potential for Clinical Utility in Patients With Melanoma

    PubMed Central

    Xu, Melody J.; Dorsey, Jay F.; Amaravadi, Ravi; Karakousis, Giorgos; Simone, Charles B.; Xu, Xiaowei; Xu, Wei; Carpenter, Erica L.; Schuchter, Lynn

    2016-01-01

    Circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), and messenger RNA (mRNA), collectively termed circulating tumor products (CTPs), represent areas of immense interest from scientists’ and clinicians’ perspectives. In melanoma, CTP analysis may have clinical utility in many areas, from screening and diagnosis to clinical decision-making aids, as surveillance biomarkers or sources of real-time genetic or molecular characterization. In addition, CTP analysis can be useful in the discovery of new biomarkers, patterns of treatment resistance, and mechanisms of metastasis development. Here, we compare and contrast CTCs, ctDNA, and mRNA, review the extent of translational evidence to date, and discuss how future studies involving both scientists and clinicians can help to further develop this tool for the benefit of melanoma patients. Implications for Practice: Scientific advancement has enabled the rapid development of tools to analyze circulating tumor cells, tumor DNA, and messenger RNA, collectively termed circulating tumor products (CTPs). A variety of techniques have emerged to detect and characterize melanoma CTPs; however, only a fraction has been applied to human subjects. This review summarizes the available human data that investigate clinical utility of CTP in cancer screening, melanoma diagnosis, prognosis, prediction, and genetic or molecular characterization. It provides a rationale for how CTPs may be useful for future research and discusses how clinicians can be involved in developing this exciting new technology. PMID:26614709

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

    PubMed

    Gopinath, Rajaneesh Karimpurath; Leu, Jun-Yi

    2016-05-01

    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.

  19. 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.

  20. Utility of poly(ethylene glycol) based hydrogels for improved neural cell culture and cell delivery systems

    NASA Astrophysics Data System (ADS)

    Namba, Rachael

    Cell transplant therapy is a very promising treatment for patients afflicted with neurodegenerative diseases and involves replacing dying neurons with fresh fetal tissue that is composed of both new neurons and multipotent neural precursor cells (NPCs). Unfortunately, this therapy is not clinically feasible given the scarcity of fresh donor fetal tissue. The NPCs present in fetal tissue may be a promising renewable cell source because they can be instructed to divide indefinitely in culture, expanding the number of NPCs that are available. Importantly, NPCs can also be instructed to differentiate into the therapeutic neurons needed to treat patients. However, when NPCs are expanded in culture, they increasingly differentiate into glial cells. Glial cell contamination not only introduces many uncharacterized signals into NSC cultures, but when transplanted into the brain, glia can contribute to inflammation and glial scarring. The goal of this project is to identify in vitro culture conditions that can be used to expand fresh NPCs while completely avoiding glial differentiation, only allowing the generation of neuronal progeny with therapeutic potential. The culture system used in this work was chosen carefully, and after exploring the utility of standard neural cell culture systems such as monolayer, neurosphere, and three-dimensional fibrin matrices, an advantageous culture system was identified. The chosen culture system is a synthetic, degradable, photo-polymerizable, poly (ethylene) glycocl (PEG) hydrogel. This thesis experimentally demonstrates advantages of PEG hydrogel culture over the other culture systems including enrichment of the initial cell population for NPCs, clonal expansion capabilities, increased potency of proliferation-stimulating mitogens, decreased reactivity and growth of glial cells, and control over the initiation and orientation of neural process growth. PEG hydrogels were used to directly assess the influence of various ECM proteins and

  1. Attachment of MAL32-encoded maltase on the outside of yeast cells improves maltotriose utilization.

    PubMed

    Dietvorst, J; Blieck, L; Brandt, R; Van Dijck, P; Steensma, H Y

    2007-01-01

    The fermentation of maltotriose, the second most abundant fermentable sugar in wort, is often incomplete during high-gravity brewing. Poor maltotriose consumption is due to environmental stress conditions during high-gravity fermentation and especially to a low uptake of this sugar by some industrial strains. In this study we investigated whether the use of strains with an alpha-glucosidase attached to the outside of the cell might be a possible way to reduce residual maltotriose. To this end, the N-terminal leader sequence of Kre1 and the carboxy-terminal anchoring domain of either Cwp2 or Flo1 were used to target maltase encoded by MAL32 to the cell surface. We showed that Mal32 displayed on the cell surface of Saccharomyces cerevisiae laboratory strains was capable of hydrolysis of alpha-1,4-linkages, and that it increased the ability of a strain lacking a functional maltose permease to grow on maltotriose. Moreover, the enzyme was also expressed and found to be active in an industrial strain. These data show that expressing a suitable maltase on the cell surface might provide a means of modifying yeast for more complete maltotriose utilization in brewing and other fermentation applications.

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. Immunocytochemistry of epithelial markers in citral-induced prostate hyperplasia in rats.

    PubMed

    Massas, R; Servadio, C; Sandbank, U; Abramovici, A

    1991-04-01

    Immunocytochemical characterization of several epithelial markers using the PAP technique was analyzed during different stages of induced prostatic hyperplasia in rats. Intact adolescent rats (42 days old) were treated with citral (3,7 dimethyl-2,6 octadienal) for 10, 30 and 100 days and their ventral prostate compared to untreated, matched-age animals. Among the epithelial markers studied the prostatic specific acid phosphatase was present in hyperplastic prostates of rats. The immunoreaction showed a fair correlation with the severity of lesion and duration of treatment. The prostatic specific antigen showed equally immunoreactive in both control and treated rats. The hyperplastic and normal rat prostates did not show immunoreactivity towards the other epithelial cell markers such as epithelial membrane antigen, carcinoembrionic antingen and alpha-fetoprotein antisera. It is concluded that prostatic specific acid phosphatase, and to a lesser extent prostatic specific antigen, might represent valuable markers for comparative studies of prostatic hyperplasia in rodents.

  7. 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

  8. 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

  9. Sex steroid-mediated reprogramming of vascular smooth muscle cells to stem cells and neurons: possible utilization of sex steroid combinations for regenerative treatment without utilization of in vitro developed stem cells.

    PubMed

    Bukovsky, Antonin

    2009-12-15

    Previous work from our laboratory demonstrated that sex steroid combinations, but not individual sex steroids alone, cause transdifferentiation of ovarian epithelial cells--ovarian surface epithelium (OSE) and follicular granulosa cells--into neural stem cells (NSC) and differentiating neurons. In the present study we have chosen primary culture of human vascular smooth muscle cells (SMC), a non-epithelial mesenchymal cells in order to test them as a control cell type regarding their morphology and expression of NSC and neuronal markers. Utilization of estradiol (E2), progesterone (PG) or testosterone (TS) alone did not induce the emergence of neurons from the vascular SMC. However, the treatment with sex steroid combinations (PG + TS or E2 + PG + TS) caused transdifferentiation into neural/neuronal type cells. By immunohistochemistry, these cells exhibited strong expression of stem cell markers and neural/neuronal glycoconjugates SSEA-1, SSEA-4, Thy-1, NeuN and NCAM. In the Neurobasal/B27 medium both, the OSE and vascular SMC also transdifferentiated into neuronal cells. Western blot analysis has shown significant increase of NeuN 48-kDa species after E2 + PG or PG + TS treatment. Secretion of E2 increased significantly in vascular SMC cultures pretreated with TS, PG or TS + PG. Unlike OSE cells, the vascular SMC accompany as pericytes all vessels, including CNS microvasculature. We also observed that sex steroid combinations could produce SMC stem type cells which differentiated within a few days back to mature vascular SMC. This is of potential interest for the vascular regenerative medicine. Altogether, our observations suggest that sex steroid combinations could induce in vivo improvement of neurodegenerative, traumatic and ischemic neurological disorders and vascular diseases via their effect on resident pluripotent vascular SMC, i.e., without a need of in vitro developed stem cells.

  10. 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.

  11. Carbon deposition thresholds on nickel-based solid oxide fuel cell anodes I. Fuel utilization

    NASA Astrophysics Data System (ADS)

    Kuhn, J.; Kesler, O.

    2015-03-01

    In the first of a two part publication, the effect of fuel utilization (Uf) on carbon deposition rates in solid oxide fuel cell nickel-based anodes was studied. Representative 5-component CH4 reformate compositions (CH4, H2, CO, H2O, & CO2) were selected graphically by plotting the solutions to a system of mass-balance constraint equations. The centroid of the solution space was chosen to represent a typical anode gas mixture for each nominal Uf value. Selected 5-component and 3-component gas mixtures were then delivered to anode-supported cells for 10 h, followed by determination of the resulting deposited carbon mass. The empirical carbon deposition thresholds were affected by atomic carbon (C), hydrogen (H), and oxygen (O) fractions of the delivered gas mixtures and temperature. It was also found that CH4-rich gas mixtures caused irreversible damage, whereas atomically equivalent CO-rich compositions did not. The coking threshold predicted by thermodynamic equilibrium calculations employing graphite for the solid carbon phase agreed well with empirical thresholds at 700 °C (Uf ≈ 32%); however, at 600 °C, poor agreement was observed with the empirical threshold of ˜36%. Finally, cell operating temperatures correlated well with the difference in enthalpy between the supplied anode gas mixtures and their resulting thermodynamic equilibrium gas mixtures.

  12. 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.

  13. Timing and dynamics of single cell gene expression in the arabinose utilization system.

    PubMed

    Megerle, Judith A; Fritz, Georg; Gerland, Ulrich; Jung, Kirsten; Rädler, Joachim O

    2008-08-01

    The arabinose utilization system of Escherichia 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. Glass-sandwich-type organic solar cells utilizing liquid crystalline phthalocyanine

    NASA Astrophysics Data System (ADS)

    Usui, Toshiki; Nakata, Yuya; De Romeo Banoukepa, Gilles; Fujita, Kento; Nishikawa, Yuki; Shimizu, Yo; Fujii, Akihiko; Ozaki, Masanori

    2017-02-01

    Glass-sandwich-type organic solar cells utilizing liquid crystalline phthalocyanine, 1,4,8,11,15,18,22,25-octahexylphthalocyanine (C6PcH2), have been fabricated and their photovoltaic properties have been studied. The short-circuit current density (J sc) and power conversion efficiency (PCE) depend on the C6PcH2 layer thickness, and the maximum performance, such as a J sc of 7.1 mA/cm2 and a PCE of 1.64%, was demonstrated for a device having a 420-nm-thick C6PcH2 layer. We examined the photovoltaic properties from the viewpoint of the C6PcH2-layer electrical conductance, based on the distribution of the column-axis direction.

  20. 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

  1. 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.

  2. 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

  3. Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures

    PubMed Central

    Morrison, Eamonn; Wai, Patty; Leonidou, Andri; Bland, Philip; Khalique, Saira; Farnie, Gillian; Daley, Frances; Peck, Barrie; Natrajan, Rachael

    2016-01-01

    The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery of the next generation of novel drug targets. It is becoming apparent that more complex models of cancer are required to fully appreciate the contributing factors that drive tumorigenesis in vivo and increase the efficacy of novel therapies that make the transition from pre-clinical models to clinical trials. Here we present a methodology for generating uniform and reproducible tumor spheroids that can be subjected to siRNA functional screening. These spheroids display many characteristics that are found in solid tumors that are not present in traditional two-dimension culture. We show that several commonly used breast cancer cell lines are amenable to this protocol. Furthermore, we provide proof-of-principle data utilizing the breast cancer cell line BT474, confirming their dependency on amplification of the epidermal growth factor receptor HER2 and mutation of phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) when grown as tumor spheroids. Finally, we are able to further investigate and confirm the spatial impact of these dependencies using immunohistochemistry. PMID:28060271

  4. Design and utilization of macrophage and vascular smooth muscle cell co-culture systems in atherosclerotic cardiovascular disease investigation.

    PubMed

    Zuniga, Mary C; White, Sharla L Powell; Zhou, Wei

    2014-10-01

    Atherosclerotic cardiovascular disease has been acknowledged as a chronic inflammatory condition. Monocytes and macrophages lead the inflammatory pathology of atherosclerosis whereas changes in atheromatous plaque thickness and matrix composition are attributed to vascular smooth muscle cells. Because these cell types are key players in atherosclerosis progression, it is crucial to utilize a reliable system to investigate their interaction. In vitro co-culture systems are useful platforms to study specific molecular mechanisms between cells. This review aims to summarize the various co-culture models that have been developed to investigate vascular smooth muscle cell and monocyte/macrophage interactions, focusing on the monocyte/macrophage effects on vascular smooth muscle cell function.

  5. A high fuel utilizing solid oxide fuel cell cycle with regard to the formation of nickel oxide and power density

    NASA Astrophysics Data System (ADS)

    Nehter, Pedro

    Within this study a novel high fuel utilizing (High-uf) SOFC system is presented with special focus on the formation of nickel oxide, system efficiency and the required cell area at a fixed system performance of 1 MW. Within the High-uf SOFC cycle, a second SOFC stack is used to utilize a further part of the residual hydrogen of the first SOFC stack. This could be feasible by using an anode gas condenser, which is implemented between the first and the second stack. This reduces the water fraction of the anode gas and thereby the tendency of nickel oxide formation in case of a further fuel utilization. Thus, a higher total fuel utilization can be reached with the second SOFC stack. With the High-uf SOFC cycle, the system efficiency is increased by 7%-points compared to the simple atmospheric SOFC cycle. Furthermore, the average cell voltage and the fuel utilization are varied to carry out a first optimization of the stack's power density. The results of this optimization have shown that the required cell area of the simple SOFC cycle can be slightly reduced by decreasing the fuel utilization, whereas the High-uf SOFC cycle shows an opposite effect. Here, the required cell area can be reduced at constant voltages by increasing the fuel utilization. Thus, higher system efficiencies could be reached with the High-uf SOFC cycle by using the same cell area as the simple SOFC cycle and at the same tendency of nickel oxide formation. A second condenser behind the second SOFC stack could be used to increase the carbon dioxide mass fraction up to 92%. This could be interesting for CO 2-sequestring applications as well.

  6. 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

  7. The Utilization during Mitotic Cell Division of Loci Controlling Meiotic Recombination and Disjunction in DROSOPHILA MELANOGASTER

    PubMed Central

    Baker, Bruce S.; Carpenter, Adelaide T. C.; Ripoll, P.

    1978-01-01

    To inquire whether the loci identified by recombination-defective and disjunction-defective meiotic mutants in Drosophila are also utilized during mitotic cell division, the effects of 18 meiotic mutants (representing 13 loci) on mitotic chromosome stability have been examined genetically. To do this, meiotic-mutant-bearing flies heterozygous for recessive somatic cell markers were examined for the frequencies and types of spontaneous clones expressing the cell markers. In such flies, marked clones can arise via mitotic recombination, mutation, chromosome breakage, nondisjunction or chromosome loss, and clones from these different origins can be distinguished. In addition, meiotic mutants at nine loci have been examined for their effects on sensitivity to killing by UV and X rays.—Mutants at six of the seven recombination-defective loci examined (mei-9, mei-41, c(3)G, mei-W68, mei-S282, mei-352, mei-218) cause mitotic chromosome instability in both sexes, whereas mutants at one locus (mei-218) do not affect mitotic chromosome stability. Thus many of the loci utilized during meiotic recombination also function in the chromosomal economy of mitotic cells.—The chromosome instability produced by mei-41 alleles is the consequence of chromosome breakage, that of mei-9 alleles is primarily due to chromosome breakage and, to a lesser extent, to an elevated frequency of mitotic recombination, whereas no predominant mechanism responsible for the instability caused by c(3)G alleles is discernible. Since these three loci are defective in their responses to mutagen damage, their effects on chromosome stability in nonmutagenized cells are interpreted as resulting from an inability to repair spontaneous lesions. Both mei-W68 and mei-S282 increase mitotic recombination (and in mei-W68, to a lesser extent, chromosome loss) in the abdomen but not the wing. In the abdomen, the primary effect on chromosome stability occurs during the larval period when the abdominal histoblasts

  8. Manipulation of human early T lymphopoiesis by coculture on human bone marrow stromal cells: potential utility for adoptive immunotherapy.

    PubMed

    Liu, Bing; Ohishi, Kohshi; Orito, Yuki; Nakamori, Yoshiki; Nishikawa, Hiroyoshi; Ino, Kazuko; Suzuki, Kei; Matsumoto, Takeshi; Masuya, Masahiro; Hamada, Hirofumi; Mineno, Junichi; Ono, Ryoichi; Nosaka, Tetsuya; Shiku, Hiroshi; Katayama, Naoyuki

    2013-04-01

    T cell precursors are an attractive target for adoptive immunotherapy. We examined the regulation of human early T lymphopoiesis by human bone marrow stromal cells to explore in vitro manipulation of human T cell precursors in a human-only coculture system. The generation of CD7(+)CD56(-)cyCD3(-) proT cells from human hematopoietic progenitors on telomerized human bone marrow stromal cells was enhanced by stem cell factor, flt3 ligand, and thrombopoietin, but these stimulatory effects were suppressed by interleukin 3. Expression of Notch ligands Delta-1 and -4 on stromal cells additively promoted T cell differentiation into the CD7(+)cyCD3(+) pre-T cell stage, while cell growth was strongly inhibited. By combining these coculture systems, we found that initial coculture with telomerized stromal cells in the presence of stem cell factor, flt3 ligand, and thrombopoietin, followed by coculture on Delta-1- and -4-coexpressing stromal cells led to a higher percentage and number of pre-T cells. Adoptive immunotherapy using peripheral blood T cells transduced with a tumor antigen-specific T cell receptor (TCR) is a promising strategy but has several limitations, such as the risk of forming a chimeric TCR with the endogenous TCR. We demonstrated that incubation of TCR-transduced hematopoietic progenitors with the combination of coculture systems gave rise to CD7(+)TCR(+)CD3(+)CD1a(-) T cell precursors that rapidly proliferated and differentiated under the culture condition to induce mature T cell differentiation. These data show the regulatory mechanism of early T lymphopoiesis on human stromal cells and the potential utility of engineered human stromal cells to manipulate early T cell development for clinical application.

  9. 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.

  10. BK virus-hemorrhagic cystitis following allogeneic stem cell transplantation: Clinical characteristics and utility of leflunomide treatment.

    PubMed

    Park, Young Hoon; Lim, Joo Han; Yi, Hyeon Gyu; Lee, Moon Hee; Kim, Chul Soo

    2016-04-18

    BK virus-hemorrhagic cystitis (BKV-HC) is a potential cause of morbidity and mortality in patients having undergone allogeneic stem cell transplantation (Allo-SCT). We analyzed the clinical features of BKV-HC following Allo-SCT and reported the utility of leflunomide therapy for BKV-HC.

  11. Permeability characteristics of novel mydriatic agents using an in vitro cell culture model that utilizes SIRC rabbit corneal cells.

    PubMed

    Goskonda, V R; Khan, M A; Hutak, C M; Reddy, I K

    1999-02-01

    The purpose of this study was to evaluate the permeability characteristics of a previously reported in vitro corneal model that utilizes SIRC rabbbit corneal cells and to investigate the permeability of three novel esters of phenylephrone chemical delivery systems (CDS) under different pH conditions using this in vitro model. The SIRC rabbit corneal cell line was grown on transwell polycarbonate membranes, and the barrier properties were assessed by measuring transepithelial electrical resistance (TEER) using a voltohmmeter. The permeabilities of esters of phenylephrone CDS across the SIRC cell layers were measured over a pH range 4.0-7. 4. The esters tested include phenylacetyl (1), isovaleryl (2), and pivalyl (3). The SIRC rabbit corneal cell line, when grown on permeable filters, formed tight monolayers of high electrical resistance with TEER values increasing from 71.6 +/- 20.8 Omega.cm2 at day 3 in culture to 2233.42 +/- 15.2 Omega.cm2 at day 8 in culture and remained constant through day 14 in culture. The transepithelial permeability coefficients (Papp) at pH 7.4 ranged from 0.58 x 10(-6) cm/s for the hydrophilic marker, mannitol, to 43. 5 x 10(-6) cm/s for the most lipophilic molecule, testosterone. The Papp at pH 7.4 for phenylephrine was 4.21 x 10(-6) cm/s. The Papp values and the lag times of the three esters of phenylephrone were pH dependent. The Papp for 1, 2, and 3 at pH 7.4 were 14.76 x 10(-6), 13.19 x 10(-6), and 12.86 x 10(-6) cm/s, respectively and the permeabilities decreased at conditions below pH 7.4. The lag times at pH 7.4 were 0.10, 0.17, and 0.12 h for 1, 2, and 3, respectively, and the values increased at lower pH conditions. The TEER values of SIRC cell line observed at day 8 to day 14 in the present investigation are similar to the resistance value reported for rabbit cornea (2 kOmega.cm2). All the esters showed significantly (p < 0.05) higher permeabilities than phenylephrine at pH 7.4. The rate and extent of transport of the drugs

  12. 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.

  13. A microchip-based endothelium mimic utilizing open reservoirs for cell immobilization and integrated carbon ink microelectrodes for detection.

    PubMed

    Hulvey, Matthew K; Martin, R Scott

    2009-01-01

    This paper describes the fabrication and characterization of a microfluidic device that utilizes a reservoir-based approach for endothelial cell immobilization and integrated embedded carbon ink microelectrodes for the amperometric detection of extracellular nitric oxide (NO) release. The design utilizes a buffer channel to continuously introduce buffer or a plug of stimulant to the reservoir as well as a separate sampling channel that constantly withdraws buffer from the reservoir and over the microelectrode. A steel pin is used for both the fluidic connection to the sampling channel and to provide a quasi-reference electrode for the carbon ink microelectrode. Characterization of the device was performed using NO standards produced from a NONOate salt. Finally, NO release from a layer of immobilized endothelial cells was monitored and quantified using the system. This system holds promise as a means to electrochemically detect extracellular NO release from endothelial cells in either an array of reservoirs or concurrently with fluorescence-based intracellular NO measurements.

  14. 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.

  15. Clinical utility of flow cytometry in the study of erythropoiesis and nonclonal red cell disorders.

    PubMed

    Chesney, Alden; Good, David; Reis, Marciano

    2011-01-01

    Erythropoiesis involves proliferation and differentiation of small population of hematopoietic stem cells resident in the bone marrow into mature red blood cells. The determination of the cellular composition of the blood is a valuable tool in the diagnosis of diseases and monitoring of therapy. Flow cytometric analysis is increasingly being used to characterize the heterogeneous cell populations present in the blood and the hematopoietic cell differentiation and maturation pathways of the bone marrow. Here we discuss the role of flow cytometry in the study of erythropoiesis and nonclonal red blood cell disorders. First, we discuss flow cytometric analysis of reticulocytes. Next, we review salient quantitative methods that can be used for detection of fetal-maternal hemorrhage (FMH). We also discuss flow cytometric analysis of high hemoglobin F (HbF) in Sickle Cell Disease (SCD), hereditary spherocytosis (HS), red cell survival and red cell volume. We conclude by discussing cell cycle of erythroid cells.

  16. 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.

  17. Clinical utilization of cord blood over human health: experience of stem cell transplantation and cell therapy using cord blood in Korea

    PubMed Central

    2014-01-01

    Cord blood (CB) has been used as an important and ethical source for hematopoietic stem cell transplantation (SCT) as well as cell therapy by manufacturing mesenchymal stem cell, induced pleuripotential stem cell or just isolating mononuclear cell from CB. Recently, the application of cell-based therapy using CB has expanded its clinical utility, particularly, by using autologous CB in children with refractory diseases. For these purposes, CB has been stored worldwide since mid-1990. In this review, I would like to briefly present the historical development of clinical uses of CB in the fields of SCT and cell therapy, particularly to review the experiences in Korea. Furthermore, I would touch the recent banking status of CB. PMID:24778692

  18. Utility of temporally distinct baculovirus promoters for constitutive and baculovirus-inducible transgene expression in transformed insect cells.

    PubMed

    Lin, Chi-Hung; Jarvis, Donald L

    2013-05-10

    Genetically transformed lepidopteran insect cell lines have biotechnological applications as constitutive recombinant protein production platforms and improved hosts for baculovirus-mediated recombinant protein production. Insect cell transformation is often accomplished with a DNA construct(s) encoding a foreign protein(s) under the transcriptional control of a baculovirus immediate early promoter, such as the ie1 promoter. However, the potential utility of increasingly stronger promoters from later baculovirus gene classes, such as delayed early (39K), late (p6.9), and very late (polh), has not been systematically assessed. Hence, we produced DNA constructs encoding secreted alkaline phosphatase (SEAP) under the transcriptional control of each of the four temporally distinct classes of baculovirus promoters, used them to transform insect cells, and compared the levels of SEAP RNA and protein production obtained before and after baculovirus infection. The ie1 construct was the only one that supported SEAP protein production by transformed insect cells prior to baculovirus infection, confirming that only immediate early promoters can be used to isolate transformed insect cells for constitutive recombinant protein production. However, baculovirus infection activated transgene expression by all four classes of baculovirus promoters. After infection, cells transformed with the very late (polh) and late (p6.9) promoter constructs produced the highest levels of SEAP RNA, but only low levels of SEAP protein. Conversely, cells transformed with the immediate early (ie1) and delayed early (39K) promoter constructs produced lower levels of RNA, but equal or higher levels of SEAP protein. Unexpectedly, the 39K promoter construct provided tightly regulated, baculovirus-inducible protein production at higher levels than the later promoter constructs. Thus, this study demonstrated the utility of the 39K promoter for insect cell engineering, particularly when one requires higher

  19. 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

    2015-10-07

    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.

  20. 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

  1. 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.

  2. Near infrared enhancement in CIGS-based solar cells utilizing a ZnO:H window layer.

    PubMed

    Yeh, Chi-Li; Hsu, Hung-Ru; Chen, Sheng-Hui; Liu, Yung-Sheng

    2012-11-05

    We investigated the near infrared enhancement in Cu(In,Ga)Se(2) (CIGS)- based solar cells utilizing a hydrogen-doping ZnO (ZnO:H) window layer. The results show that the carrier concentration of ZnO:H film is lower than that of AZO film which can increase the transmittance in the NIR. The advantage of ZnO:H film is higher Hall mobility than AZO film. Thus ZnO:H film has similar resistivity to AZO film. It was found that the cell efficiency was 12.4 and 13% for the AZO device and the ZnO:H device, respectively. The cell efficiency is enhanced by 4.8%. Furthermore, the results indicate that, the ZnO:H film is superior to the AZO film as the window layer for CIGS-based solar cells.

  3. 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

  4. Pluronic F127 as a cell encapsulation material: utilization of membrane-stabilizing agents.

    PubMed

    Khattak, Sarwat F; Bhatia, Surita R; Roberts, Susan C

    2005-01-01

    Thermoreversible gelation of the copolymer Pluronic F127 (generic name, poloxamer 407) in water makes it a unique candidate for cell encapsulation applications, either alone or to promote cell seeding and attachment in tissue scaffolds. At concentrations of 15-20% (w/w), aqueous Pluronic F127 (F127) solutions gel at physiological temperatures. The effect of F127 on viability and proliferation of human liver carcinoma cells (HepG2) was determined for both liquid and gel formulations. Cell concentration and viability over a 5-day period were measured by the trypan blue assay via hemocytometry and results were confirmed in both the MTT and LDH assays. With 0.1-5% (w/w) F127 (liquid), cells proliferated and maintained high viability over 5 days. However, at 10% (w/w) F127 (liquid), there was a significant decrease in cell viability and no cell proliferation was evident. HepG2 cell encapsulation in F127 concentrations ranging from 15 to 20% (w/w) (gel) resulted in complete cell death by 5 days. This was also true for the HMEC-1 (endothelial) and L6 (muscle) cell lines evaluated. Cell-seeding density did not affect cell survival or proliferation. Membrane-stabilizing agents (hydrocortisone, glucose, and glycerol) were added to the F127 gel formulations to improve cell viability. The steroid hydrocortisone demonstrated the most significant improvement in viability, from <2% (in F127 alone) to >70% (with 60 nM hydrocortisone added). These results suggest that F127 formulations supplemented with membrane-stabilizing agents can serve as viable cell encapsulation materials. In addition, hydrocortisone may be generally useful in the promotion of cell viability for a wide range of encapsulation materials.

  5. Murine Th17 cells utilize IL-2 receptor gamma chain cytokines but are resistant to cytokine withdrawal-induced apoptosis.

    PubMed

    Neitzke, Daniel J; Bowers, Jacob S; Andrijauskaite, Kristina; O'Connell, Nathaniel S; Garrett-Mayer, Elizabeth; Wrangle, John; Li, Zihai; Paulos, Chrystal M; Cole, David J; Rubinstein, Mark P

    2017-03-09

    Adoptive cellular therapy (ACT) with the Th17 subset of CD4(+) T cells can cure established melanoma in preclinical models and holds promise for treating human cancer. However, little is known about the growth factors necessary for optimal engraftment and anti-tumor activity of Th17 cells. Due to the central role of IL-2 receptor gamma chain (IL2Rγ-chain) cytokines (IL-2, IL-7, and IL-15) in the activity and persistence of many T cell subsets after adoptive transfer, we hypothesized that these cytokines are important for Th17 cells. We found that Th17 cells proliferated in response to IL-2, IL-7, and IL-15 in vitro. However, in contrast to many other T cell subsets, including conventionally activated CD8(+) T cells, we found that Th17 cells were resistant to apoptosis in the absence of IL2Rγ-chain cytokines. To determine whether Th17 cells utilize IL2Rγ-chain cytokines in vivo, we tracked Th17 cell engraftment after adoptive transfer with or without cytokine depletion. Depletion of IL-7 and/or IL-2 decreased initial engraftment, while depletion of IL-15 did not. Supplementation of IL-2 increased initial Th17 engraftment. To assess the clinical relevance of these findings, we treated melanoma-bearing mice with Th17 cell adoptive transfer and concurrent cytokine depletion or supplementation. We found that simultaneous depletion of IL-2 and IL-7 decreased therapeutic efficacy, depletion of IL-15 had no effect, and IL-2 supplementation increased therapeutic efficacy. Our results show that Th17 cells are responsive to IL2Rγ-chain cytokines, and provide insight into the application of these cytokines for Th17-based therapeutic strategies.

  6. Improved drug targeting of cancer cells by utilizing actively targetable folic acid-conjugated albumin nanospheres.

    PubMed

    Shen, Zheyu; Li, Yan; Kohama, Kazuhiro; Oneill, Brian; Bi, Jingxiu

    2011-01-01

    Folic acid-conjugated albumin nanospheres (FA-AN) have been developed to provide an actively targetable drug delivery system for improved drug targeting of cancer cells with reduced side effects. The nanospheres were prepared by conjugating folic acid onto the surface of albumin nanospheres using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) as a catalyst. To test the efficacy of these nanospheres as a potential delivery platform, doxorubicin-loaded albumin nanospheres (DOX-AN) and doxorubicin-loaded FA-AN (FA-DOX-AN) were prepared by entrapping DOX (an anthracycline, antibiotic drug widely used in cancer chemotherapy that works by intercalating DNA) into AN and FA-AN nanoparticles. Cell uptake of the DOX was then measured. The results show that FA-AN was incorporated into HeLa cells (tumor cells) only after 2.0h incubation, whereas HeLa cells failed to incorporate albumin nanospheres without conjugated folic acid after 4.0h incubation. When HeLa cells were treated with the DOX-AN, FA-DOX-AN nanoparticles or free DOX, cell viability decreased with increasing culture time (i.e. cell death increases with time) over a 70h period. Cell viability was always the lowest for free DOX followed by FA-DOX-AN4 and then DOX-AN. In a second set of experiments, HeLa cells washed to remove excess DOX after an initial incubation for 2h were incubated for 70h. The corresponding cell viability was slightly higher when the cells were treated with FA-DOX-AN or free DOX whilst cells treated with DOX-AN nanoparticles remained viable. The above experiments were repeated for non-cancerous, aortic smooth muscle cells (AoSMC). As expected, cell viability of the HeLa cells (with FA receptor alpha, FRα) and AoSMC cells (without FRα) decreased rapidly with time in the presence of free DOX, but treatment with FA-DOX-AN resulted in selective killing of the tumor cells. These results indicated that FA-AN may be used as a promising actively targetable drug delivery system to improve drug

  7. Radiation survival of murine and human melanoma cells utilizing two assay systems: monolayer and soft agar.

    PubMed Central

    Yohem, K. H.; Slymen, D. J.; Bregman, M. D.; Meyskens, F. L.

    1988-01-01

    The radiation response of murine and human melanoma cells assayed in bilayer soft agar and monolayer was examined. Cells from the murine melanoma Cloudman S91 CCL 53.1 cell line and three human melanoma cell strains (C8146C, C8161, and R83-4) developed in our laboratory were irradiated by single dose X-rays and plated either in agar or on plastic. D0 values were the same within 95% confidence intervals for cells from the human melanoma cell strains C8146C, C8161, and R83-4 but were dissimilar for the murine cell line CCL 53.1 Dq values were different for all cells studied. The shape of the survival curve for all four melanomas was not identical for cells assayed in soft agar versus cells grown on plastic. This would indicate that apparent radiosensitivity was influenced by the method of assay although there were no apparent consistent differences between the curves generated by monolayer or bilayer soft agar assays. PMID:3348949

  8. Keratocyte fragments and cells utilize competing pathways to move in opposite directions in an electric field.

    PubMed

    Sun, Yaohui; Do, Hao; Gao, Jing; Zhao, Ren; Zhao, Min; Mogilner, Alex

    2013-04-08

    Sensing of an electric field (EF) by cells-galvanotaxis-is important in wound healing [1], development [2], cell division, nerve growth, and angiogenesis [3]. Different cell types migrate in opposite directions in EFs [4], and the same cell can switch the directionality depending on conditions [5]. A tug-of-war mechanism between multiple signaling pathways [6] can direct Dictyostelium cells to either cathode or anode. Mechanics of motility is simplest in fish keratocytes, so we turned to keratocytes to investigate their migration in EFs. Keratocytes sense electric fields and migrate to the cathode [7, 8]. Keratocyte fragments [9, 10] are the simplest motile units. Cell fragments from leukocytes are able to respond to chemotactic signals [11], but whether cell fragments are galvanotactic was unknown. We found that keratocyte fragments are the smallest motile electric field-sensing unit: they migrate to the anode, in the opposite direction of whole cells. Myosin II was essential for the direction sensing of fragments but not for parental cells, while PI3 kinase was essential for the direction sensing of whole cells but not for fragments. Thus, two signal transduction pathways, one depending on PI3K, another on myosin, compete to orient motile cells in the electric field. Galvanotaxis is not due to EF force and does not depend on cell or fragment size. We propose a "compass" model according to which protrusive and contractile actomyosin networks self-polarize to the front and rear of the motile cell, respectively, and the electric signal orients both networks toward cathode with different strengths.

  9. 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.

  10. Utilization of similar mechanisms by Legionella pneumophila to parasitize two evolutionarily distant host cells, mammalian macrophages and protozoa.

    PubMed Central

    Gao, L Y; Harb, O S; Abu Kwaik, Y

    1997-01-01

    The Legionnaires' disease bacterium, Legionella pneumophila, is an intracellular pathogen of humans that is amplified in the environment by intracellular multiplication within protozoa. Within both evolutionarily distant hosts, the bacterium multiplies in a rough endoplasmic reticulum-surrounded phagosome that is retarded from maturation through the endosomal-lysosomal degradation pathway. To gain an understanding of the mechanisms utilized by L. pneumophila to invade and replicate within two evolutionarily distant hosts, we isolated a collection of 89 mini-Tn10::kan insertion mutants that exhibited defects in cytotoxicity, intracellular survival, and replication within both U937 macrophage-like cells and Acanthamoeba polyphaga. Interestingly, the patterns of defects in intracellular survival and replication of the mutants within both host cells were highly similar, and thus we designated the defective loci in these mutants pmi (for protozoan and macrophage infectivity loci). On the basis of their ability to attach to host cells and their growth kinetics during the intracellular infection, the mutants were grouped into five groups. Groups 1 and 2 included 41 mutants that were severely defective in intracellular survival and were completely or substantially killed during the first 4 h of infection in both host cells. Three members of group 1 were severely defective in attachment to both U937 cells and A. polyphaga, and another four mutants of group 1 exhibited severe defects in attachment to A. polyphaga but only a mild reduction in their attachment to U937 cells. Four members of groups 1 and 2 were serum sensitive. Intracellular replication of mutants of the other three groups was less defective than that of mutants of groups 1 and 2, and their growth kinetics within both host cells were similar. The mutants were tested for several other phenotypes in vitro, revealing that 14 of the pmi mutants were resistant to NaCl, 3 had insertions in dot or icm, 3 were

  11. 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.

  12. Functional EpoR Pathway Utilization Is Not Detected in Primary Tumor Cells Isolated from Human Breast, Non-Small Cell Lung, Colorectal, and Ovarian Tumor Tissues

    PubMed Central

    Patterson, Scott D.; Rossi, John M.; Paweletz, Katherine L.; Fitzpatrick, V. Dan; Begley, C. Glenn; Busse, Leigh; Elliott, Steve; McCaffery, Ian

    2015-01-01

    Several clinical trials in oncology have reported increased mortality or disease progression associated with erythropoiesis-stimulating agents. One hypothesis proposes that erythropoiesis-stimulating agents directly stimulate tumor proliferation and/or survival through cell-surface receptors. To test this hypothesis and examine if human tumors utilize the erythropoietin receptor pathway, the response of tumor cells to human recombinant erythropoietin was investigated in disaggregated tumor cells obtained from 186 patients with colorectal, breast, lung, ovarian, head and neck, and other tumors. A cocktail of well characterized tumor growth factors (EGF, HGF, and IGF-1) were analyzed in parallel as a positive control to determine whether freshly-isolated tumor cells were able to respond to growth factor activation ex vivo. Exposing tumor cells to the growth factor cocktail resulted in stimulation of survival and proliferation pathways as measured by an increase in phosphorylation of the downstream signaling proteins AKT and ERK. In contrast, no activation by human recombinant erythropoietin was observed in isolated tumor cells. Though tumor samples exhibited a broad range of cell-surface expression of EGFR, c-Met, and IGF-1R, no cell-surface erythropoietin receptor was detected in tumor cells from the 186 tumors examined (by flow cytometry or Western blot). Erythropoiesis-stimulating agents did not act directly upon isolated tumor cells to stimulate pathways known to promote proliferation or survival of human tumor cells isolated from primary and metastatic tumor tissues. PMID:25807104

  13. 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.

  14. Solid oxide fuel cell bi-layer anode with gadolinia-doped ceria for utilization of solid carbon fuel

    NASA Astrophysics Data System (ADS)

    Kellogg, Isaiah D.; Koylu, Umit O.; Dogan, Fatih

    Pyrolytic carbon was used as fuel in a solid oxide fuel cell (SOFC) with a yttria-stabilized zirconia (YSZ) electrolyte and a bi-layer anode composed of nickel oxide gadolinia-doped ceria (NiO-GDC) and NiO-YSZ. The common problems of bulk shrinkage and emergent porosity in the YSZ layer adjacent to the GDC/YSZ interface were avoided by using an interlayer of porous NiO-YSZ as a buffer anode layer between the electrolyte and the NiO-GDC primary anode. Cells were fabricated from commercially available component powders so that unconventional production methods suggested in the literature were avoided, that is, the necessity of glycine-nitrate combustion synthesis, specialty multicomponent oxide powders, sputtering, or chemical vapor deposition. The easily-fabricated cell was successfully utilized with hydrogen and propane fuels as well as carbon deposited on the anode during the cyclic operation with the propane. A cell of similar construction could be used in the exhaust stream of a diesel engine to capture and utilize soot for secondary power generation and decreased particulate pollution without the need for filter regeneration.

  15. 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.

  16. A high pressure optical cell utilizing single crystal cubic zirconia anvil windows

    NASA Astrophysics Data System (ADS)

    Russell, T. P.; Piermarini, G. J.

    1997-04-01

    A high pressure optical cell capable of producing pressures up to 13.2 GPa using gem-cut single crystal cubic zirconia (CZ) anvils was developed. Maximum pressures obtainable were found to depend upon the particular pressure transmitting medium and gasket material employed. The cubic zirconia anvil high pressure cell (CZAC) provides advantages over the diamond anvil cell in optical and infrared spectroscopy while still maintaining a substantial pressure capability. To demonstrate these advantages, microRaman, optical fluorescence, and infrared absorption measurements were made on diamond, ruby, and 1,3,5-trinitrohexahydro-1,3,5-triazine samples, respectively, using the CZAC cell under high pressure conditions.

  17. 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.

  18. 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.

  19. Patient-Derived Xenograft Models of Non-Small Cell Lung Cancer and Their Potential Utility in Personalized Medicine.

    PubMed

    Morgan, Katherine M; Riedlinger, Gregory M; Rosenfeld, Jeffrey; Ganesan, Shridar; Pine, Sharon R

    2017-01-01

    Traditional preclinical studies of cancer therapeutics have relied on the use of established human cell lines that have been adapted to grow in the laboratory and, therefore, may deviate from the cancer they were meant to represent. With the emphasis of cancer drug development shifting from non-specific cytotoxic agents to rationally designed molecularly targeted therapies or immunotherapy comes the need for better models with predictive value regarding therapeutic activity and response in clinical trials. Recently, the diversity and accessibility of immunodeficient mouse strains has greatly enhanced the production and utility of patient-derived xenograft (PDX) models for many tumor types, including non-small cell lung cancer (NSCLC). Combined with next-generation sequencing, NSCLC PDX mouse models offer an exciting tool for drug development and for studying targeted therapies while utilizing patient samples with the hope of eventually aiding in clinical decision-making. Here, we describe NSCLC PDX mouse models generated by us and others, their ability to reflect the parental tumors' histomorphological characteristics, as well as the effect of clonal selection and evolution on maintaining genomic integrity in low-passage PDXs compared to the donor tissue. We also raise vital questions regarding the practical utility of PDX and humanized PDX models in predicting patient response to therapy and make recommendations for addressing those questions. Once collaborations and standardized xenotransplantation and data management methods are established, NSCLC PDX mouse models have the potential to be universal and invaluable as a preclinical tool that guides clinical trials and standard therapeutic decisions.

  20. Potential Utility and Limitations of Thyroid Cancer Cell Lines as Models for Studying Thyroid Cancer

    PubMed Central

    Pilli, Tania; Prasad, Kanteti V.; Jayarama, Shankar; Pacini, Furio

    2009-01-01

    Background Tumor-derived cell lines are widely used to study the mechanisms involved in thyroid carcinogenesis but recent studies have reported redundancy among thyroid cancer cell lines and identification of some “thyroid cell lines” that are likely not of thyroid origin. Summary In this review, we have summarized the uses, the limitations, and the existing problems associated with the available follicular cell-derived thyroid cancer cell lines. There are some limitations to the use of cell lines as a model to “mimic” in vivo tumors. Based on the gene expression profiles of thyroid cell lines originating from tumors of different types it has become apparent that some of the cell lines are closely related to each other and to those of undifferentiated carcinomas. Further, many cell lines have lost the expression of thyroid-specific genes and have altered karyotypes, while they exhibit activation of several oncogenes (BRAF, v-raf murine sarcoma viral oncogene homolog B1; RAS, rat sarcoma; and RET/PTC, rearranged in transformation/papillary thyroid carcinoma) and inactivation of tumor suppressor gene (TP53) which is known to be important for thyroid tumorigenesis. Conclusions A careful selection of thyroid cancer cell lines that reflect the major characteristics of a particular type of thyroid cancer being investigated could be used as a good model system to analyze the signaling pathways that may be important in thyroid carcinogenesis. Further, the review of literature also suggests that some of the limitations can be overcome by using multiple cell lines derived from the same type of tumor. PMID:20001716

  1. 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.

  2. 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.

  3. 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.

  4. Progression of cutaneous plasmacytoma to plasma cell leukemia in a dog.

    PubMed

    Rout, Emily D; Shank, Alba Maria M; Waite, Angharad H K; Siegel, Andrea; Avery, Anne C; Avery, Paul R

    2017-02-10

    A 5-year-old male neutered Bernese Mountain Dog was presented for cutaneous plasmacytoma, which was treated by surgical excision. Four months later, the dog developed multiple skin masses, hyphema, pericardial and mild bicavitary effusions, myocardial masses, and marked plasmacytosis in the peripheral blood. Circulating plasma cells expressed CD34 and MHC class II by flow cytometry. Immunocytochemistry demonstrated that these cells were strongly positive for multiple myeloma oncogene 1/interferon regulatory factor 4 (MUM-1) and weakly to moderately positive for Pax5. The dog was hypoglobulinemic but had a monoclonal IgA gammopathy detected by serum immunofixation electrophoresis. The PCR analysis of antigen receptor gene rearrangements (PARR) by fragment analysis using GeneScan methodology revealed that plasmacytoid cells in the original cutaneous plasmacytoma and peripheral blood had an identical immunoglobulin heavy chain gene (IgH) rearrangement, indicating that both populations were derived from the same neoplastic clone. Canine cutaneous plasmacytoma rarely progresses to a malignant form and plasma cell leukemia is rarely diagnosed in the dog. This report describes a case of cutaneous plasmacytoma progressing to plasma cell leukemia with a rapid and aggressive clinical course. This report also highlights the utility of flow cytometry, immunocytochemistry, immunofixation electrophoresis, and PARR by fragment analysis using GeneScan methodology in the diagnosis of this hematopoietic neoplasm.

  5. 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)

  6. 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.

  7. 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.

  8. Parainfluenza virus isolation enhancement utilizing a portable cell culture system in the field.

    PubMed Central

    Parkinson, A J; Muchmore, H G; Scott, L V; Miles, J A

    1980-01-01

    Using a portable minaturized cell culture system, enhanced recoveries of parainfluenza virus types 1 and 3 were made in the field from symptomatic human adult subjects working at remote Antarctic stations. PMID:6247369

  9. Utilization of graphene electrode in transparent microwell arrays for high throughput cell trapping and lysis.

    PubMed

    Ameri, S Kabiri; Singh, P K; Sonkusale, S

    2014-11-15

    Here we present a high-throughput, transparent microfluidic device with embedded microwell arrays sandwiched between transparent electrodes made from graphene (at the bottom) and indium tin oxide (at the top) for dielectrophoretic cell trapping and electrical lysis. Graphene suppresses unwanted faradaic reaction effects on the cells and the medium that is typically observed in ITO based electrodes from application of DC field for electrical lysis. This is because graphene is more electrochemically inert than indium tin oxide (ITO) where ITO undergoes reduction-oxidation (redox) reaction in the presence of electrolyte in most standard cell media. This redox process also compromises ITO's electrical properties and optical transparency over multiple use. The presented microfluidic device shows high efficiency for cell trapping and lysis and an electrochemically stable behavior for long operational life.

  10. SARS coronavirus, but not human coronavirus NL63, utilizes cathepsin L to infect ACE2-expressing cells.

    PubMed

    Huang, I-Chueh; Bosch, Berend Jan; Li, Fang; Li, Wenhui; Lee, Kyoung Hoa; Ghiran, Sorina; Vasilieva, Natalya; Dermody, Terence S; Harrison, Stephen C; Dormitzer, Philip R; Farzan, Michael; Rottier, Peter J M; Choe, Hyeryun

    2006-02-10

    Viruses require specific cellular receptors to infect their target cells. Angiotensin-converting enzyme 2 (ACE2) is a cellular receptor for two divergent coronaviruses, SARS coronavirus (SARS-CoV) and human coronavirus NL63 (HCoV-NL63). In addition to hostcell receptors, lysosomal cysteine proteases are required for productive infection by some viruses. Here we show that SARS-CoV, but not HCoV-NL63, utilizes the enzymatic activity of the cysteine protease cathepsin L to infect ACE2-expressing cells. Inhibitors of cathepsin L blocked infection by SARS-CoV and by a retrovirus pseudotyped with the SARS-CoV spike (S) protein but not infection by HCoV-NL63 or a retrovirus pseudotyped with the HCoV-NL63 S protein. Expression of exogenous cathepsin L substantially enhanced infection mediated by the SARS-CoV S protein and by filovirus GP proteins but not by the HCoV-NL63 S protein or the vesicular stomatitis virus G protein. Finally, an inhibitor of endosomal acidification had substantially less effect on infection mediated by the HCoV-NL63 S protein than on that mediated by the SARS-CoV S protein. Our data indicate that two coronaviruses that utilize a common receptor nonetheless enter cells through distinct mechanisms.

  11. The utility of yeast as a tool for cell-based, target-directed high-throughput screening.

    PubMed

    Norcliffe, J L; Alvarez-Ruiz, E; Martin-Plaza, J J; Steel, P G; Denny, P W

    2014-01-01

    Many Neglected Tropical Diseases (NTDs) have recently been subject of increased focus, particularly with relation to high-throughput screening (HTS) initiatives. These vital endeavours largely rely of two approaches, in vitro target-directed screening using biochemical assays or cell-based screening which takes no account of the target or targets being hit. Despite their successes both of these approaches have limitations; for example, the production of soluble protein and a lack of cellular context or the problems and expense of parasite cell culture. In addition, both can be challenging to miniaturize for ultra (u)HTS and expensive to utilize. Yeast-based systems offer a cost-effective approach to study and screen protein targets in a direct-directed manner within a eukaryotic cellular context. In this review, we examine the utility and limitations of yeast cell-based, target-directed screening. In particular we focus on the currently under-explored possibility of using such formats in uHTS screening campaigns for NTDs.

  12. 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

  13. Organic Nitrogen Utilization by Phytoplankton: The Role of Cell-Surface Deaminases

    DTIC Science & Technology

    1989-06-01

    inhibited by cell-impermeable protein Marler 1966: Zika 1984). This highly re- modification reagents. A cell-surface redox en- active species may have a...from wet and dry deposition ( Zika sured in cultures of I. carterae could be et al. 1982; Thompson and Zafiriou 1983), generalized, marine phytoplankton...would be an is generated in photochemical reactions important sourceofthe hydrogen peroxide found (Cooper and Zika 1983), and can also be in the manne

  14. Cell segmentation in histopathological images with deep learning algorithms by utilizing spatial relationships.

    PubMed

    Hatipoglu, Nuh; Bilgin, Gokhan

    2017-02-28

    In many computerized methods for cell detection, segmentation, and classification in digital histopathology that have recently emerged, the task of cell segmentation remains a chief problem for image processing in designing computer-aided diagnosis (CAD) systems. In research and diagnostic studies on cancer, pathologists can use CAD systems as second readers to analyze high-resolution histopathological images. Since cell detection and segmentation are critical for cancer grade assessments, cellular and extracellular structures should primarily be extracted from histopathological images. In response, we sought to identify a useful cell segmentation approach with histopathological images that uses not only prominent deep learning algorithms (i.e., convolutional neural networks, stacked autoencoders, and deep belief networks), but also spatial relationships, information of which is critical for achieving better cell segmentation results. To that end, we collected cellular and extracellular samples from histopathological images by windowing in small patches with various sizes. In experiments, the segmentation accuracies of the methods used improved as the window sizes increased due to the addition of local spatial and contextual information. Once we compared the effects of training sample size and influence of window size, results revealed that the deep learning algorithms, especially convolutional neural networks and partly stacked autoencoders, performed better than conventional methods in cell segmentation.

  15. 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.

  16. A mechanistic dissection of polyethylenimine mediated transfection of CHO cells: to enhance the efficiency of recombinant DNA utilization.

    PubMed

    Mozley, Olivia L; Thompson, Ben C; Fernandez-Martell, Alejandro; James, David C

    2014-01-01

    In this study, we examine the molecular and cellular interactions that underpin efficient internalization and utilization of polyethylenimine (PEI):DNA complexes (polyplexes) by Chinese Hamster Ovary (CHO) cells. Cell surface polyplex binding and internalization was a biphasic process, consisting of an initial rapid Phase (I), lasting approximately 15 min, followed by a slower second Phase (II), saturating at approximately 240 min post transfection. The second Phase accounted for the majority (60-70%) of polyplex internalization. While cell surface heparan sulphate proteoglycans (HSPGs) were rapidly cointernalized with polyplexes during Phase I, cell surface polyplex binding was not dependent on HSPGs. However, Phase II polyplex internalization and HSPG regeneration onto the surface of trypsinized cells occurred at similar rates, suggesting that the rate of recycling of HSPG-containing membrane to the plasma membrane limits Phase II internalization rate. Under optimal transfection conditions, polyplexes had a near neutral surface charge (zeta potential) and cell surface binding was dependent on hydrophobic interactions, being significantly inhibited by both chemical sequestration of cholesterol from the plasma membrane and addition of nonionic surfactant. Induced alterations in polyplex zeta potential, using ferric (III) citrate to decrease surface charge and varying PEI:DNA ratio to increase surface charge, served to inhibit polyplex binding or reduce secreted alkaline phosphatase reporter expression and cell viability, respectively. To increase polyplex hydrophobicity and internalization an alkylated derivative of PEI, propyl-PEI, was chemically synthesized. Using Design of Experiments-Response Surface Modeling to optimize the transfection process, the function of propyl-PEI was compared to that of unmodified PEI in both parental CHO-S cells and a subclone (Clone 4), which exhibited superior transgene expression via an increased resistance to polyplex

  17. TGF-beta 1 stimulation of cell locomotion utilizes the hyaluronan receptor RHAMM and hyaluronan

    PubMed Central

    1993-01-01

    TGF-beta is a potent stimulator of motility in a variety of cell types. It has recently been shown that hyaluronan (HA) can directly promote locomotion of cells through interaction with the HA receptor RHAMM. We have investigated the role of RHAMM and HA in TGF-beta-stimulated locomotion and show that TGF-beta triggers the transcription, synthesis and membrane expression of the RHAMM receptor and the secretion of HA coincident with the induction of the locomotory response. This was demonstrated by both incubating cells with exogenous TGF-beta 1 and by stimulating the production of bioactive TGF-beta 1 in tumor cells transfected with TGF-beta 1 under the control of the metallothionein promoter. TGF-beta 1-induced locomotion was suppressed by antibodies that prevented HA/RHAMM interaction, using polyclonal antibodies to either RHAMM fusion protein or RHAMM peptides, or mAbs to purified RHAMM. Peptides corresponding to the HA-binding motif of RHAMM also suppressed TGF-beta 1-induced increases in motility rate. Spontaneous locomotion of fibrosarcoma cells was blocked by neutralizing secreted TGF-beta with panspecific TGF-beta antibodies and by inhibition of TGF- beta 1 secretion with antisense oligonucleotides. Polyclonal anti-RHAMM fusion protein antibodies and peptide from the RHAMM HA-binding motif also suppressed the spontaneous motility rate of fibrosarcoma cells. These data suggest that fibrosarcoma cell locomotion requires TGF-beta, and the pathway by which TGF-beta stimulates locomotion uses the HA receptor RHAMM and HA. PMID:7693717

  18. Re-utilization of Schwann cells during ingrowth of ventral root afferents in perinatal kittens.

    PubMed

    Nilsson Remahl, A Ingela M; Masterman, Thomas; Risling, Mårten

    2008-08-01

    Ventral roots in all mammalian species, including humans, contain significant numbers of unmyelinated axons, many of them afferents transmitting nociceptive signals from receptive fields in skin, viscera, muscles and joints. Observations in cats indicate that these afferents do not enter the spinal cord via the ventral root, but rather turn distally and enter the dorsal root. Some unmyelinated axons are postganglionic autonomic efferents that innervate blood vessels of the root and the pia mater. In the feline L7 segment, a substantial proportion of unmyelinated axons are not detectable until late in perinatal development. The mechanisms inducing this late ingrowth, and the recruitment of Schwann cells (indispensable, at this stage, for axonal survival and sustenance), are unknown. We have counted axons and Schwann cells in both ends of the L7 ventral root in young kittens and made the following observations. (1) The total number of axons detectable in the root increased throughout the range of investigated ages. (2) The number of myelinated axons was similar in the root's proximal and distal ends. The increased number of unmyelinated axons with age is thus due to increased numbers of small unmyelinated axons. (3) The number of separated large probably promyelin axons was about the same in the proximal and distal ends of the root. (4) Schwann cells appeared to undergo redistribution, from myelinated to unmyelinated axons. (5) During redistribution of Schwann cells they first appear as aberrant Schwann cells and then become endoneurial X-cells temporarily free of axonal contact. We hypothesize that unmyelinated axons invade the ventral root from its distal end, that this ingrowth is particularly intense during the first postnatal month and that disengaged Schwann cells, eliminated from myelinated motoneuron axons, provide the ingrowing axons with structural and trophic support.

  19. 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.

  20. TGF-beta 1 stimulation of cell locomotion utilizes the hyaluronan receptor RHAMM and hyaluronan.

    PubMed

    Samuel, S K; Hurta, R A; Spearman, M A; Wright, J A; Turley, E A; Greenberg, A H

    1993-11-01

    TGF-beta is a potent stimulator of motility in a variety of cell types. It has recently been shown that hyaluronan (HA) can directly promote locomotion of cells through interaction with the HA receptor RHAMM. We have investigated the role of RHAMM and HA in TGF-beta-stimulated locomotion and show that TGF-beta triggers the transcription, synthesis and membrane expression of the RHAMM receptor and the secretion of HA coincident with the induction of the locomotory response. This was demonstrated by both incubating cells with exogenous TGF-beta 1 and by stimulating the production of bioactive TGF-beta 1 in tumor cells transfected with TGF-beta 1 under the control of the metallothionein promoter. TGF-beta 1-induced locomotion was suppressed by antibodies that prevented HA/RHAMM interaction, using polyclonal antibodies to either RHAMM fusion protein or RHAMM peptides, or mAbs to purified RHAMM. Peptides corresponding to the HA-binding motif of RHAMM also suppressed TGF-beta 1-induced increases in motility rate. Spontaneous locomotion of fibrosarcoma cells was blocked by neutralizing secreted TGF-beta with panspecific TGF-beta antibodies and by inhibition of TGF-beta 1 secretion with antisense oligonucleotides. Polyclonal anti-RHAMM fusion protein antibodies and peptide from the RHAMM HA-binding motif also suppressed the spontaneous motility rate of fibrosarcoma cells. These data suggest that fibrosarcoma cell locomotion requires TGF-beta, and the pathway by which TGF-beta stimulates locomotion uses the HA receptor RHAMM and HA.

  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. 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.

  3. System for exposing cultured cells to intermittent hypoxia utilizing gas permeable cultureware.

    PubMed

    Polak, Jan; Studer-Rabeler, Karen; McHugh, Holly; Hussain, Mehboob A; Shimoda, Larissa A

    2015-07-01

    Tissue intermittent hypoxia (IH) occurs in obstructive sleep apnea, sickle cell anemia, physical exercise and other conditions. Poor gas solubility and slow diffusion through culture media hampers mimicking IH-induced transitions of O(2) in vitro. We aimed to develop a system enabling exposure of cultured cells to IH and to validate such exposure by real-time O(2) measurements and cellular responses. Standard 24-well culture plates and plates with bottoms made from a gas permeable film were placed in a heated cabinet. Desired cycling of O(2) levels was induced using programmable solenoids to purge mixtures of 95% N(2) + 5% CO(2) or 95% O(2) + 5% CO(2). Dissolved oxygen, gas pressure, temperature, and water evaporation were measured during cycling. IH-induced cellular effects were evaluated by hypoxia inducible factor (HIF) and NF-κB luciferase reporters in HEK296 cells and by insulin secretion in rat insulinoma cells. Oxygen cycling in the cabinet was translated into identical changes of O(2) at the well bottom in gas permeable, but not in standard cultureware. Twenty-four hours of IH exposure increased HIF (112%), NF-κB (111%) and insulin secretion (44%). Described system enables reproducible and prolonged IH exposure in cultured cells while controlling for important environmental factors.

  4. 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.

  5. 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.

  6. Diagnostic utility of immunohistochemistry in distinguishing trichoepithelioma and basal cell carcinoma: evaluation using tissue microarray samples.

    PubMed

    Tebcherani, Antonio José; de Andrade, Heitor Franco; Sotto, Mirian N

    2012-10-01

    Trichoepithelioma is a benign neoplasm that shares both clinical and histological features with basal cell carcinoma. It is important to distinguish these neoplasms because they require different clinical behavior and therapeutic planning. Many studies have addressed the use of immunohistochemistry to improve the differential diagnosis of these tumors. These studies present conflicting results when addressing the same markers, probably owing to the small number of basaloid tumors that comprised their studies, which generally did not exceed 50 cases. We built a tissue microarray with 162 trichoepithelioma and 328 basal cell carcinoma biopsies and tested a panel of immune markers composed of CD34, CD10, epithelial membrane antigen, Bcl-2, cytokeratins 15 and 20 and D2-40. The results were analyzed using multiple linear and logistic regression models. This analysis revealed a model that could differentiate trichoepithelioma from basal cell carcinoma in 36% of the cases. The panel of immunohistochemical markers required to differentiate between these tumors was composed of CD10, cytokeratin 15, cytokeratin 20 and D2-40. The results obtained in this work were generated from a large number of biopsies and resulted in the confirmation of overlapping epithelial and stromal immunohistochemical profiles from these basaloid tumors. The results also corroborate the point of view that trichoepithelioma and basal cell carcinoma tumors represent two different points in the differentiation of a single cell type. Despite the use of panels of immune markers, histopathological criteria associated with clinical data certainly remain the best guideline for the differential diagnosis of trichoepithelioma and basal cell carcinoma.

  7. Translational efficiency of polycistronic mRNAs and their utilization to express heterologous genes in mammalian cells.

    PubMed Central

    Kaufman, R J; Murtha, P; Davies, M V

    1987-01-01

    The translation of polycistronic mRNAs in mammalian cells was studied. Transcription units, constructed to contain one, two or three open reading frames (ORFs), were introduced stably into Chinese hamster ovary cells and transiently into COS monkey cells. The analysis of mRNA levels and protein synthesis in these cells demonstrated that the mRNAs transcribed were translated to generate multiple proteins. The efficiency of translation was reduced approximately 40- to 300-fold by the insertion of an upstream ORF. The results support a modified 'scanning' model for translation initiation which allows for translation initiation at internal AUG codons. High-level expression of human granulocyte-macrophage colony stimulating factor was achieved utilizing a vector that contains a polycistronic transcription unit encoding an amplifiable dihydrofolate reductase marker gene in its 3' end. Thus, polycistronic expression vectors can be exploited to obtain high-level expression of foreign genes in mammalian cells. Images Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. Fig. 8. PMID:3582359

  8. 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.

  9. 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.

  10. Biosynthesis of 14C-phytoene from tomato cell suspension cultures (Lycopersicon esculentum) for utilization in prostate cancer cell culture studies.

    PubMed

    Campbell, Jessica K; Rogers, Randy B; Lila, Mary Ann; Erdman, John W

    2006-02-08

    This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.

  11. Utilizing pharmacotherapy and mesenchymal stem cell therapy to reduce inflammation following traumatic brain injury

    PubMed Central

    Mashkouri, Sherwin; Crowley, Marci G.; Liska, Michael G.; Corey, Sydney; Borlongan, Cesar V.

    2016-01-01

    The pathologic process of chronic phase traumatic brain injury is associated with spreading inflammation, cell death, and neural dysfunction. It is thought that sequestration of inflammatory mediators can facilitate recovery and promote an environment that fosters cellular regeneration. Studies have targeted post-traumatic brain injury inflammation with the use of pharmacotherapy and cell therapy. These therapeutic options are aimed at reducing the edematous and neurodegenerative inflammation that have been associated with compromising the integrity of the blood-brain barrier. Although studies have yielded positive results from anti-inflammatory pharmacotherapy and cell therapy individually, emerging research has begun to target inflammation using combination therapy. The joint use of anti-inflammatory drugs alongside stem cell transplantation may provide better clinical outcomes for traumatic brain injury patients. Despite the promising results in this field of research, it is important to note that most of the studies mentioned in this review have completed their studies using animal models. Translation of this research into a clinical setting will require additional laboratory experiments and larger preclinical trials. PMID:27857726

  12. Genetically Targeted Radiotherapy Utilizing the Human Sodium Iodide Symporter in Human Breast Carcinoma Cells

    DTIC Science & Technology

    2007-07-01

    several histone deactylase inhibitors ( HDACi ), including sodium butyrate (SB), trichostatin A (TSA), in conjunction with the DNA methyltransferase...breast cancer cell lines and increase the tumor response, several ne deactylase inhibitors ( HDACi ), were tested. The HDACi including sodium butyrate (SB

  13. Production of D-tagatose, a functional sweetener, utilizing alginate immobilized Lactobacillus fermentum CGMCC2921 cells.

    PubMed

    Xu, Zheng; Li, Sha; Fu, Fenggen; Li, Guixiang; Feng, Xiaohai; Xu, Hong; Ouyang, Pingkai

    2012-02-01

    D-tagatose is a ketohexose that can be used as a novel functional sweetener in foods, beverages, and dietary supplements. This study was aimed at developing a high-yielding D-tagatose production process using alginate immobilized Lactobacillus fermentum CGMCC2921 cells. For the isomerization from D-galactose into D-tagatose, the immobilized cells showed optimum temperature and pH at 65 °C and 6.5, respectively. The alginate beads exhibited a good stability after glutaraldehyde treatment and retained 90% of the enzyme activity after eight cycles (192 h at 65 °C) of batch conversion. The addition of borate with a molar ratio of 1.0 to D-galactose led to a significant enhancement in the D-tagatose yield. Using commercial β-galactosidase and immobilized L. fermentum cells, D-tagatose was successfully obtained from lactose after a two-step biotransformation. The relatively high conversion rate and productivity from D-galactose to D-tagatose of 60% and 11.1 g l⁻¹ h⁻¹ were achieved in a packed-bed bioreactor. Moreover, lactobacilli have been approved as generally recognized as safe organisms, which makes this L. fermentum strain an attracting substitute for recombinant Escherichia coli cells among D-tagatose production progresses.

  14. 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

  15. 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.…

  16. Utilizing insulating nanoparticles as the spacer in laminated flexible polymer solar cells for improved mechanical stability.

    PubMed

    Lu, Yunzhang; Alexander, Clement; Xiao, Zhengguo; Yuan, Yongbo; Zhang, Runyu; Huang, Jinsong

    2012-08-31

    Roll-to-roll lamination is one promising technique to produce large-area organic electronic devices such as solar cells with a large through output. One challenge in this process is the frequent electric point shorting of the cathode and anode by the excess or concentrated applied stress from many possible sources. In this paper, we report a method to avoid electric point shorting by incorporating insulating and hard barium titanate (BaTiO(3)) nanoparticles (NPs) into the active layer to work as a spacer. It has been demonstrated that the incorporated BaTiO(3) NPs in poly(3-hexylthiophene):[6,6]-phenyl-c-61-butyric acid methyl ester (P3HT:PCBM) bulk heterojunction solar cells cause no deleterious effect to the power conversion process of this type of solar cell. The resulting laminated devices with NPs in the active layer display the same efficiency as the devices without NPs, while the laminated devices with NPs can sustain a ten times higher lamination stress of over 6 MPa. The flexible polymer solar cell device with incorporated NPs shows a much smaller survivable curvature radius of 4 mm, while a regular flexible device can only sustain a bending curvature radius of 8 mm before fracture.

  17. Utilizing insulating nanoparticles as the spacer in laminated flexible polymer solar cells for improved mechanical stability

    NASA Astrophysics Data System (ADS)

    Lu, Yunzhang; Alexander, Clement; Xiao, Zhengguo; Yuan, Yongbo; Zhang, Runyu; Huang, Jinsong

    2012-08-01

    Roll-to-roll lamination is one promising technique to produce large-area organic electronic devices such as solar cells with a large through output. One challenge in this process is the frequent electric point shorting of the cathode and anode by the excess or concentrated applied stress from many possible sources. In this paper, we report a method to avoid electric point shorting by incorporating insulating and hard barium titanate (BaTiO3) nanoparticles (NPs) into the active layer to work as a spacer. It has been demonstrated that the incorporated BaTiO3 NPs in poly(3-hexylthiophene):[6,6]-phenyl-c-61-butyric acid methyl ester (P3HT:PCBM) bulk heterojunction solar cells cause no deleterious effect to the power conversion process of this type of solar cell. The resulting laminated devices with NPs in the active layer display the same efficiency as the devices without NPs, while the laminated devices with NPs can sustain a ten times higher lamination stress of over 6 MPa. The flexible polymer solar cell device with incorporated NPs shows a much smaller survivable curvature radius of 4 mm, while a regular flexible device can only sustain a bending curvature radius of 8 mm before fracture.

  18. 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

  19. Helicobacter pylori Salvages Purines from Extracellular Host Cell DNA Utilizing the Outer Membrane-Associated Nuclease NucT

    PubMed Central

    Liechti, George W.

    2013-01-01

    Helicobacter pylori is a bacterial pathogen that establishes life-long infections in humans, and its presence in the gastric epithelium is strongly associated with gastritis, peptic ulcer disease, and gastric cancer. Having evolved in this specific gastric niche for hundreds of thousands of years, this microbe has become dependent on its human host. Bioinformatic analysis reveals that H. pylori has lost several genes involved in the de novo synthesis of purine nucleotides, and without this pathway present, H. pylori must salvage purines from its environment in order to grow. While the presence and abundance of free purines in various mammalian tissues has been loosely quantified, the concentration of purines present within the gastric mucosa remains unknown. There is evidence, however, that a significant amount of extracellular DNA is present in the human gastric mucosal layer as a result of epithelial cell turnover, and this DNA has the potential to serve as an adequate purine source for gastric purine auxotrophs. In this study, we characterize the ability of H. pylori to grow utilizing only DNA as a purine source. We show that this ability is independent of the ComB DNA uptake system, and that H. pylori utilization of DNA as a purine source is largely influenced by the presence of an outer membrane-associated nuclease (NucT). A ΔnucT mutant exhibits significantly reduced extracellular nuclease activity and is deficient in growth when DNA is provided as the sole purine source in laboratory growth media. These growth defects are also evident when this nuclease mutant is grown in the presence of AGS cells or in purine-free tissue culture medium that has been conditioned by AGS cells in the absence of fetal bovine serum. Taken together, these results indicate that the salvage of purines from exogenous host cell DNA plays an important role in allowing H. pylori to meet its purine requirements for growth. PMID:23893109

  20. 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

  1. Patient-Derived Xenograft Models of Non-Small Cell Lung Cancer and Their Potential Utility in Personalized Medicine

    PubMed Central

    Morgan, Katherine M.; Riedlinger, Gregory M.; Rosenfeld, Jeffrey; Ganesan, Shridar; Pine, Sharon R.

    2017-01-01

    Traditional preclinical studies of cancer therapeutics have relied on the use of established human cell lines that have been adapted to grow in the laboratory and, therefore, may deviate from the cancer they were meant to represent. With the emphasis of cancer drug development shifting from non-specific cytotoxic agents to rationally designed molecularly targeted therapies or immunotherapy comes the need for better models with predictive value regarding therapeutic activity and response in clinical trials. Recently, the diversity and accessibility of immunodeficient mouse strains has greatly enhanced the production and utility of patient-derived xenograft (PDX) models for many tumor types, including non-small cell lung cancer (NSCLC). Combined with next-generation sequencing, NSCLC PDX mouse models offer an exciting tool for drug development and for studying targeted therapies while utilizing patient samples with the hope of eventually aiding in clinical decision-making. Here, we describe NSCLC PDX mouse models generated by us and others, their ability to reflect the parental tumors’ histomorphological characteristics, as well as the effect of clonal selection and evolution on maintaining genomic integrity in low-passage PDXs compared to the donor tissue. We also raise vital questions regarding the practical utility of PDX and humanized PDX models in predicting patient response to therapy and make recommendations for addressing those questions. Once collaborations and standardized xenotransplantation and data management methods are established, NSCLC PDX mouse models have the potential to be universal and invaluable as a preclinical tool that guides clinical trials and standard therapeutic decisions. PMID:28154808

  2. 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

  3. 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.

  4. Canine distemper virus utilizes different receptors to infect chicken embryo fibroblasts and vero cells.

    PubMed

    Chen, Jun; Liang, Xiu; Chen, Pei-fu

    2011-04-01

    Inducing animal viruses to adapt to chicken embryos or chicken embryo fibroblasts (CEF) is a common method to develop attenuated live vaccines with full security. Canine distemper virus (CDV) also does this, but the mechanisms and particular receptors remain unclear. Virus overlay protein blot assays were carried out on CEF membrane proteins, which were extracted respectively with a Mem-PER™ kit, a radioimmunoprecipitation assay buffer or a modified co-immunoprecipitation method, and revealed a common 57 kDa positive band that differed from the 42-kDa positive band in Vero cells and also from those receptors reported in lymphocytes and 293 cells, indicating a receptor diversity of CDV and the possibility of the 57-kDa protein acting as a receptor that is involved in adaptive infection of CDV Kunming strain to CEF.

  5. 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.

  6. Development of Adaptive Tilt Tracker that Utilizes QUAD-cell Detector to Track Extended Objects

    DTIC Science & Technology

    2014-03-17

    such that the dynamic range of the tilt mirror and sensor is kept relatively “small”. This further translates to compact controller...effective QUAD-cell dimensions to 1 mm x 1 mm where the image has ample dynamic range to move when under the influence of tilt disturbance. The PTS...identify a range of Gaussian spot spreads and corresponding centroid gains that most accurately track tilt distortion over a given dynamic range of

  7. 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.

  8. Functional genomics screening utilizing mutant mouse embryonic stem cells identifies novel radiation-response genes.

    PubMed

    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.

  9. 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

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

    PubMed

    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

    2015-05-01

    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.

  11. Sonidegib: mechanism of action, pharmacology, and clinical utility for advanced basal cell carcinomas

    PubMed Central

    Jain, Sachin; Song, Ruolan; Xie, Jingwu

    2017-01-01

    The Hedgehog (Hh) pathway is critical for cell differentiation, tissue polarity, and stem cell maintenance during embryonic development, but is silent in adult tissues under normal conditions. However, aberrant Hh signaling activation has been implicated in the development and promotion of certain types of cancer, including basal cell carcinoma (BCC), medulloblastoma, and gastrointestinal cancers. In 2015, the US Food and Drug Administration (FDA) approved sonidegib, a smoothened (SMO) antagonist, for treatment of advanced BCC (aBCC) after a successful Phase II clinical trial. Sonidegib, also named Odomzo, is the second Hh signaling inhibitor approved by the FDA to treat BCCs following approval of the first SMO antagonist vismodegib in 2012. What are the major features of sonidegib (mechanism of action; metabolic profiles, clinical efficacy, safety, and tolerability profiles)? Will the sonidegib experience help other clinical trials using Hh signaling inhibitors in the future? In this review, we will summarize current understanding of BCCs and Hh signaling. We will focus on sonidegib and its use in the clinic, and we will discuss ways to improve its clinical application in cancer therapeutics. PMID:28352196

  12. Biodiesel Production: Utilization of Loofah Sponge to Immobilize Rhizopus chinensis CGMCC #3.0232 Cells as a Whole-Cell Biocatalyst.

    PubMed

    He, Qiyang; Xia, Qianjun; Wang, Yuejiao; Li, Xun; Zhang, Yu; Hu, Bo; Wang, Fei

    2016-07-28

    Rhizopus chinensis cells immobilized on loofah (Luffa cylindrica) sponges were used to produce biodiesel via the transesterification of soybean oil. In whole-cell immobilization, loofah sponge is considered to be a superior alternative to conventional biomass carriers because of its biodegradable and renewable properties. During cell cultivation, Rhizopus chinensis mycelia can spontaneously and firmly adhere to the surface of loofah sponge particles. The optimal conditions for processing 9.65 g soybean oil at 40°C and 180 rpm using a 3:1 methanol-to-oil molar ratio were found to be 8% cell addition and 3-10% water content (depending on the oil's weight). Under optimal conditions, an over 90% methyl ester yield was achieved after the first reaction batch. The operational stability of immobilized Rhizopus chinensis cells was assayed utilizing a 1:1 methanol-to-oil molar ratio, thus resulting in a 16.5-fold increase in half-life when compared with immobilized cells of the widely studied Rhizopus oryzae. These results suggest that transesterification of vegetable oil using Rhizopus chinensis whole cells immobilized onto loofah sponge is an effective approach for biodiesel production.

  13. 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.

  14. On-chip fabrication of alkali-metal vapor cells utilizing an alkali-metal source tablet

    NASA Astrophysics Data System (ADS)

    Tsujimoto, K.; Ban, K.; Hirai, Y.; Sugano, K.; Tsuchiya, T.; Mizutani, N.; Tabata, O.

    2013-11-01

    We describe a novel on-chip microfabrication technique for the alkali-metal vapor cell of an optically pumped atomic magnetometer (OPAM), utilizing an alkali-metal source tablet (AMST). The newly proposed AMST is a millimeter-sized piece of porous alumina whose considerable surface area holds deposited alkali-metal chloride (KCl) and barium azide (BaN6), source materials that effectively produce alkali-metal vapor at less than 400 °C. Our experiments indicated that the most effective pore size of the AMST is between 60 and 170 µm. The thickness of an insulating glass spacer holding the AMST was designed to confine generated alkali metal to the interior of the vapor cell during its production, and an integrated silicon heater was designed to seal the device using a glass frit, melted at an optimum temperature range of 460-490 °C that was determined by finite element method thermal simulation. The proposed design and AMST were used to successfully fabricate a K cell that was then operated as an OPAM with a measured sensitivity of 50 pT. These results demonstrate that the proposed concept for on-chip microfabrication of alkali-metal vapor cells may lead to effective replacement of conventional glassworking approaches.

  15. Utilizing of Adsorptive Transfer Stripping Technique Brdicka Reaction for Determination of Metallothioneins Level in Melanoma Cells, Blood Serum and Tissues.

    PubMed

    Krizkova, Sona; Fabrik, Ivo; Adam, Vojtech; Kukacka, Jiri; Prusa, Richard; Chavis, Grace J; Trnkova, Libuse; Strnadel, Jan; Horak, Vratislav; Kizek, Rene

    2008-05-10

    In the paper we utilized the adsorptive transfer stripping differential pulse voltammetry Brdicka reaction for the determination of metallothioneins (MT) in melanoma cells, animal melanoma tissues (MeLiM miniature pig) and blood serum of patients with malignant melanoma. Primarily we attempted to investigate the influence of dilution of real sample on MT electrochemical response. Dilution of samples of 1 000 times was chosen the most suitable for determination of MT level in biological samples. Then we quantified the MT level in the melanoma cells, the animal melanoma tissues and the blood serum samples. The MT content in the cells varied within the range from 4.2 to 11.2 μM. At animal melanoma tissues (melanomas localized on abdomen, back limb and dorsum) the highest content of MT was determined in the tumour sampled on the back of the animal and was nearly 500 μg of MTs per gram of a tissue. We also quantified content of MT in metastases, which was found in liver, spleen and lymph nodes. Moreover the average MT level in the blood serum samples from patients with melanoma was 3.0 ± 0.8 μM. MT levels determined at melanoma samples were significantly (p < 0.05) higher compared to control ones at cells, tissues and blood serum.

  16. Utilizing of Adsorptive Transfer Stripping Technique Brdicka Reaction for Determination of Metallothioneins Level in Melanoma Cells, Blood Serum and Tissues

    PubMed Central

    Krizkova, Sona; Fabrik, Ivo; Adam, Vojtech; Kukacka, Jiri; Prusa, Richard; Chavis, Grace J.; Trnkova, Libuse; Strnadel, Jan; Horak, Vratislav; Kizek, Rene

    2008-01-01

    In the paper we utilized the adsorptive transfer stripping differential pulse voltammetry Brdicka reaction for the determination of metallothioneins (MT) in melanoma cells, animal melanoma tissues (MeLiM miniature pig) and blood serum of patients with malignant melanoma. Primarily we attempted to investigate the influence of dilution of real sample on MT electrochemical response. Dilution of samples of 1 000 times was chosen the most suitable for determination of MT level in biological samples. Then we quantified the MT level in the melanoma cells, the animal melanoma tissues and the blood serum samples. The MT content in the cells varied within the range from 4.2 to 11.2 μM. At animal melanoma tissues (melanomas localized on abdomen, back limb and dorsum) the highest content of MT was determined in the tumour sampled on the back of the animal and was nearly 500 μg of MTs per gram of a tissue. We also quantified content of MT in metastases, which was found in liver, spleen and lymph nodes. Moreover the average MT level in the blood serum samples from patients with melanoma was 3.0 ± 0.8 μM. MT levels determined at melanoma samples were significantly (p < 0.05) higher compared to control ones at cells, tissues and blood serum. PMID:27879868

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

    PubMed

    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

  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. 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.

  20. 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.

  1. 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.

  2. 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

  3. Spatially and Temporally Resolved Single-Cell Exocytosis Utilizing Individually Addressable Carbon Microelectrode Arrays

    PubMed Central

    Zhang, Bo; Adams, Kelly L.; Luber, Sarah J.; Eves, Daniel J.; Heien, Michael L.; Ewing, Andrew G.

    2009-01-01

    We report the fabrication and characterization of carbon microelectrode arrays (MEAs) and their application to spatially and temporally resolve neurotransmitter release from single pheochromocytoma (PC12) cells. The carbon MEAs are composed of individually addressable 2.5-μm-radius microdisks embedded in glass. The fabrication involves pulling a multibarrel glass capillary containing a single carbon fiber in each barrel into a sharp tip, followed by beveling the electrode tip to form an array (10−20 μm) of carbon microdisks. This simple fabrication procedure eliminates the need for complicated wiring of the independent electrodes, thus allowing preparation of high-density individually addressable microelectrodes. The carbon MEAs have been characterized using scanning electron microscopy, steady-state and fast-scan voltammetry, and numerical simulations. Amperometric results show that subcellular heterogeneity in single-cell exocytosis can be electrochemically detected with MEAs. These ultrasmall electrochemical probes are suitable for detecting fast chemical events in tight spaces, as well as for developing multifunctional electrochemical microsensors. PMID:18232712

  4. 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.

  5. 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.

  6. Review on utilization of the pervaporation membrane for passive vapor feed direct methanol fuel cell

    NASA Astrophysics Data System (ADS)

    Fauzi, N. F. I.; Hasran, U. A.; Kamarudin, S. K.

    2013-12-01

    The Direct Methanol Fuel Cell (DMFC) is a promising portable power source for mobile electronic devices because of its advantages including easy fuel storage, high energy density, low temperature operation and compact structure. In DMFC, methanol is used as a fuel source where it can be fed in liquid or vapor phase. However, the vapor feed DMFC has an advantage over the liquid feed system as it has the potential to have a higher operating temperature to increase the reaction rates and power outputs, to enhance the mass transfers, to reduce methanol crossover, reliable for high methanol concentration and it can increase the fuel cell performance. Methanol vapor can be delivered to the anode by using a pervaporation membrane, heating the liquid methanol or another method that compatible. Therefore, this paper is a review on vapor feed DMFC as a better energy source than liquid feed DMFC, the pervaporation membrane used to vaporize methanol feed from the reservoir and its applications in vapor feed DMFC.

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

    DOE PAGES

    Collette, R.; Douglas, J.; Patterson, L.; ...

    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

  8. 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

  9. Environmental and economic aspects of hydrogen production and utilization in fuel cell vehicles

    NASA Astrophysics Data System (ADS)

    Granovskii, Mikhail; Dincer, Ibrahim; Rosen, Marc A.

    A smooth transition from gasoline-powered internal combustion engine vehicles to ecologically clean hydrogen fuel cell vehicles depends on the process used for hydrogen production. Three technologies for hydrogen production are considered here: traditional hydrogen production via natural gas reforming, and the use of two renewable technologies (wind and solar electricity generation) to produce hydrogen via water electrolysis. It is shown that a decrease of environmental impact (air pollution and greenhouse gas emissions) as a result of hydrogen implementation as a fuel is accompanied by a decline in the economic efficiency (as measured by capital investments effectiveness). A mathematical procedure is proposed to obtain numerical estimates of environmental and economic criteria interactions in the form of sustainability indexes. On the basis of the obtained sustainability indexes, it is concluded that hydrogen production from wind energy via electrolysis is more advantageous for mitigating greenhouse gas emissions and traditional natural gas reforming is more favorable for reducing air pollution.

  10. Thylakoid direct photobioelectrocatalysis: utilizing stroma thylakoids to improve bio-solar cell performance.

    PubMed

    Rasmussen, Michelle; Minteer, Shelley D

    2014-08-28

    Thylakoid membranes from spinach were separated into grana and stroma thylakoid fractions which were characterized by several methods (pigment content, protein gel electrophoresis, photosystem activities, and electron microscopy analysis) to confirm that the intact thylakoids were differentiated into the two domains. The results of photoelectrochemical experiments showed that stroma thylakoid electrodes generate photocurrents more than four times larger than grana thylakoids (51 ± 4 nA cm(-2) compared to 11 ± 1 nA cm(-2)). A similar trend was seen in a bio-solar cell configuration with stroma thylakoids giving almost twice the current (19 ± 3 μA cm(-2)) as grana thylakoids (11 ± 2 μA cm(-2)) with no change in open circuit voltage.

  11. 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

  12. One-step, low-temperature deposited perovskite solar cell utilizing small molecule additive

    NASA Astrophysics Data System (ADS)

    Chen, Chun-Chao; Hong, Zirou; Li, Gang; Chen, Qi; Zhou, Huanping; Yang, Yang

    2015-01-01

    In the current study, the perovskite absorber (CH3NH3PbI3) is processed via one-step deposition employing the small molecule additive, BmPyPhB, which can be dissolved in dimethylformamide along with precursors. Here, 1,3-Bis[3,5-di(pyridin-3-yl)phenyl]benzene (BmPyPhB) functions as the morphology controller to introduce an intermediate phase during perovskite film growth, which allows well-defined and precrystallized domains formed before the annealing treatment. Furthermore, a chloroform solvent wash procedure is applied afterward to remove BmPyPhB from perovskite without damaging the predetermined morphology. Thus, postannealing as low as 100°C for 5 min can achieve the optimal power conversion efficiency of 8% in a planar-structured inverted solar cell.

  13. Embedded fiber-optic sensing for accurate internal monitoring of cell state in advanced battery management systems part 2: Internal cell signals and utility for state estimation

    NASA Astrophysics Data System (ADS)

    Ganguli, Anurag; Saha, Bhaskar; Raghavan, Ajay; Kiesel, Peter; Arakaki, Kyle; Schuh, Andreas; Schwartz, Julian; Hegyi, Alex; Sommer, Lars Wilko; Lochbaum, Alexander; Sahu, Saroj; Alamgir, Mohamed

    2017-02-01

    A key challenge hindering the mass adoption of Lithium-ion and other next-gen chemistries in advanced battery applications such as hybrid/electric vehicles (xEVs) has been management of their functional performance for more effective battery utilization and control over their life. Contemporary battery management systems (BMS) reliant on monitoring external parameters such as voltage and current to ensure safe battery operation with the required performance usually result in overdesign and inefficient use of capacity. More informative embedded sensors are desirable for internal cell state monitoring, which could provide accurate state-of-charge (SOC) and state-of-health (SOH) estimates and early failure indicators. Here we present a promising new embedded sensing option developed by our team for cell monitoring, fiber-optic (FO) sensors. High-performance large-format pouch cells with embedded FO sensors were fabricated. This second part of the paper focuses on the internal signals obtained from these FO sensors. The details of the method to isolate intercalation strain and temperature signals are discussed. Data collected under various xEV operational conditions are presented. An algorithm employing dynamic time warping and Kalman filtering was used to estimate state-of-charge with high accuracy from these internal FO signals. Their utility for high-accuracy, predictive state-of-health estimation is also explored.

  14. Reptilian reovirus utilizes a small type III protein with an external myristylated amino terminus to mediate cell-cell fusion.

    PubMed

    Corcoran, Jennifer A; Duncan, Roy

    2004-04-01

    Reptilian reovirus is one of a limited number of nonenveloped viruses that are capable of inducing cell-cell fusion. A small, hydrophobic, basic, 125-amino-acid fusion protein encoded by the first open reading frame of a bicistronic viral mRNA is responsible for this fusion activity. Sequence comparisons to previously characterized reovirus fusion proteins indicated that p14 represents a new member of the fusion-associated small transmembrane (FAST) protein family. Topological analysis revealed that p14 is a representative of a minor subset of integral membrane proteins, the type III proteins N(exoplasmic)/C(cytoplasmic) (N(exo)/C(cyt)), that lack a cleavable signal sequence and use an internal reverse signal-anchor sequence to direct membrane insertion and protein topology. This topology results in the unexpected, cotranslational translocation of the essential myristylated N-terminal domain of p14 across the cell membrane. The topology and structural motifs present in this novel reovirus membrane fusion protein further accentuate the diversity and unusual properties of the FAST protein family and clearly indicate that the FAST proteins represent a third distinct class of viral membrane fusion proteins.

  15. [Utility of 8-colours multiparameter flow cytometry immunophenotyping of plasma cells for the management of monoclonal gammopathy].

    PubMed

    Gressier, Mélanie; Chaquin, Michael; Lhermitte, Ludovic; Asnafi, Vahid; Macintyre, Elizabeth; Brouzes, Chantal

    2013-01-01

    Bone marrow flow cytometric analysis is a powerful and rapid tool for evaluating aberrant plasma cell. In this study, we have examined the utility of multiparameter flow cytometry (MFC) in 52 patients with multiple myeloma (MM) and in 45 patients with monoclonal gammopathy with unknown significance (MGUS) into routine evaluation for the management of patients with plasma cell-related disorders. The plasma cells (PC) were identified by their light scatter distribution and reactivity patterns to CD138, CD38, and CD45. The combination of these parameters was helpful for identifying distinct subpopulations of PCs. Moderate to bright expression of CD56, CD20, CD24, CD28, and CD117 was detected in 67%, 26%, 13%, 27%, and 57% of MM cases and in 58%, 20%, 11%, 43% and 44% of MGUS cases, respectively. In MGUS group, the median percentage abnormal PCs/total PCs was 88% with 37 patients out of 45 (82%) with ratio <95%. The median ratio of the MM group was 98.9% and a ratio ≥ 95% was observed in 37 samples out of 44 (84%). In conclusion, MFC immunophenotyping of PCs has obvious clinical relevance in differential diagnosis between MM and others monoclonal gammopathies, identification of high-risk MGUS and smouldering MM, and minimal residual disease monitoring of MM. Our results showed that this tool can be easily applied in haematology laboratories.

  16. 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.

  17. 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.

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

    PubMed Central

    Wang, Yingchun; 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

  19. Recent advances in the development and utilization of modern anode materials for high performance microbial fuel cells.

    PubMed

    Sonawane, Jayesh M; Yadav, Abhishek; Ghosh, Prakash C; Adeloju, Samuel B

    2017-04-15

    Microbial fuel cells (MFCs) are novel bio-electrochemical device for spontaneous or single step conversion of biomass into electricity, based on the use of metabolic activity of bacteria. The design and use of MFCs has attracted considerable interests because of the potential new opportunities they offer for sustainable production of energy from biodegradable and reused waste materials. However, the associated slow microbial kinetics and costly construction materials has limited a much wider commercial use of the technology. In the past ten years, there has been significant new developments in MFCs which has resulted in several-fold increase in achievable power density. Yet, there is still considerable possibility for further improvement in performance and development of new cost effective materials. This paper comprehensively reviews recent advances in the construction and utilization of novel anodes for MFCs. In particular, it highlights some of the critical roles and functions of anodes in MFCs, strategies available for improving surface areas of anodes, dominant performance of stainless-steel based anode materials, and the emerging benefits of inclusion of nanomaterials. The review also demonstrates that some of the materials are very promising for large scale MFC applications and are likely to replace conventional anodes for the development of next generation MFC systems. The hurdles to the development of commercial MFC technology are also discussed. Furthermore, the future directions in the design and selection of materials for construction and utilization of MFC anodes are highlighted.

  20. 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.

  1. 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.

  2. Safety and Utility of Quantitative Sensory Testing among Adults with Sickle Cell Disease: Indicators of Neuropathic Pain?

    PubMed Central

    Ezenwa, Miriam O.; Molokie, Robert E.; Wang, Zaijie Jim; Yao, Yingwei; Suarez, Marie L.; Pullum, Cherese; Schlaeger, Judith M.; Fillingim, Roger B.; Wilkie, Diana J.

    2014-01-01

    Objectives Pain is the hallmark symptom of sickle cell disease (SCD), yet the types of pain that these patients experience, and the underlying mechanisms, have not been well characterized. The study purpose was to determine the safety and utility of a mechanical and thermal quantitative sensory testing (QST) protocol and the feasibility of utilizing neuropathic pain questionnaires among adults with SCD. Methods A convenience sample (N=25, 18 women, mean age 38.5 ± 12.5 [20–58 years]) completed self-report pain and quality-of-life tools. Subjects also underwent testing with the TSA-II NeuroSensory Analyzer and calibrated von Frey microfilaments. Results We found that the QST protocol was safe and did not stimulate a SCD pain crisis. There was evidence of central sensitization (n=15), peripheral sensitization (n=1), a mix of central and peripheral sensitization (n=8), or no sensitization (n=1). The neuropathic pain self-report tools were feasible with evidence of construct validity; 40% of the subjects reported S-LANSS scores that were indicative of neuropathic pain and had evidence of central, peripheral or mixed sensitization. Discussion The QST protocol can be safely conducted in adults with SCD and provides evidence of central or peripheral sensitization, which is consistent with a neuropathic component to SCD pain. These findings are novel, warrant a larger confirmatory study, and indicate the need for normative QST data from African American adults and older adults. PMID:25581383

  3. Platelet and red blood cell utilization and transfusion independence in umbilical cord blood and allogeneic peripheral blood hematopoietic cell transplants.

    PubMed

    Solh, Melhem; Brunstein, Claudio; Morgan, Shanna; Weisdorf, Daniel

    2011-05-01

    Allogeneic hematopoietic cell transplantation (HCT) recipients have substantial transfusion requirements. Factors associated with increased transfusions and the extent of blood product use in umbilical cord blood (UCB) recipients are uncertain. We reviewed blood product use in 229 consecutive adult recipients of allogeneic HCT at the University of Minnesota: 147 with leukemia, 82 lymphoma or myeloma; 58% received unrelated UCB and 43% sibling donor peripheral blood stem cell (PBSC) grafts. Although neutrophil recovery was prompt (UCB median 17, range: 2-45 days, and PBSC 14, range: 3-34 days), only 135 of 229 (59% cumulative incidence) achieved red blood cell (RBC) independence and 157 (69%) achieved platelet independence by 6 months. Time to platelet independence was prolonged in UCB recipients (median UCB 41 versus PBSC 14 days) and in patients who had received a prior transplant (median 48 versus 32 days). Patients who received UCB grafts required more RBC through day 60 post-HCT (mean UCB 7.8 (95% confidence interval [CI] 6.7-8.9) versus PBSC 5.2 (3.7-6.7) transfusions, P = .04), and more platelet transfusions (mean 25.2 (95% CI 22.1-28.2) versus 12.9 (9.4-16.4), P < .01) compared to PBSC recipients. Patients receiving myeloablative (MA) conditioning required more RBC and platelet transfusions during the first 2 months post-HCT compared to reduced-intensity conditioning (RIC) (7.4 versus 6.2, P = .30 for RBC; 23.2 versus 17.5, P = .07 for platelets). Despite prompt neutrophil engraftment, UCB recipients had delayed platelet recovery as well as more prolonged and costly blood product requirements. Enhanced approaches to accelerate multilineage engraftment could limit the transfusion-associated morbidity and costs accompanying UCB allotransplantation.

  4. The clinical utility of the proliferating cell nuclear antigen expression in patients with hepatocellular carcinoma.

    PubMed

    Ma, Shuangshuang; Yang, Junsheng; Li, Jinpeng; Song, Jinlong

    2016-06-01

    Proliferating cell nuclear antigen (PCNA) has been suggested as a potential diagnostic biomarker for early hepatocellular carcinoma (HCC). However, its prognostic significance in HCC remains unclear. In the present study, we investigated the expression and significance of PCNA in HCC and then analyzed the role of PCNA in clinical outcomes. Our findings show that the expression intensity of PCNA is much higher in HCC tissues than that in paracarcinoma tissues and associated with AFP, albumin, tumor number, clinical grade, vascular invasion, and tumor-node-metastasis (TNM) stage (all p < 0.000). Kaplan-Meier analysis indicated that high PCNA expression was associated with poor disease-free survival (DFS) (p < 0.000) and overall survival (OS) (p < 0.000) in a training cohort of 76 HCC patients. Multiple Cox regression analysis indicated PCNA acts as an independent predictor for DFS (p = 0.002) and OS (p = 0.004) in HCC patients. Along with pathological results, our systematic review also identified the expression of PCNA was closely associated with DFS and OS (both p < 0.000). In conclusion, this study suggested that PCNA is increased in HCC patients and is indeed a novel unfavorable biomarker for prognostic prediction for patients with this deadly disease.

  5. 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 .

  6. Models of endometriosis and their utility in studying progression to ovarian clear cell carcinoma.

    PubMed

    King, Claire M; Barbara, Cynthia; Prentice, Andrew; Brenton, James D; Charnock-Jones, D Stephen

    2016-01-01

    Endometriosis is a common benign gynaecological condition affecting at least 10% of women of childbearing age and is characterized by pain--frequently debilitating. Although the exact prevalence is unknown, the economic burden is substantial (∼$50 billion a year in the USA alone) and it is associated with considerable morbidity. The development of endometriosis is inextricably linked to the process of menstruation and thus the models that best recapitulate the human disease are in menstruating non-human primates. However, the use of these animals is ethically challenging and very expensive. A variety of models in laboratory animals have been developed and the most recent are based on generating menstrual-like endometrial tissue that can be transferred to a recipient animal. These models are genetically manipulable and facilitate precise mechanistic studies. In addition, these models can be used to study malignant transformation in epithelial ovarian carcinoma. Epidemiological and molecular evidence indicates that endometriosis is the most plausible precursor of both clear cell and endometrioid ovarian cancer (OCCA and OEA, respectively). While this progression is rare, understanding the underlying mechanisms of transformation may offer new strategies for prevention and therapy. Our ability to pursue this is highly dependent on improved animal models but the current transgenic models, which genetically modify the ovarian surface epithelium and oviduct, are poor models of ectopic endometrial tissue. In this review we describe the various models of endometriosis and discuss how they may be applicable to developing our mechanistic understanding of OCCA and OEA.

  7. 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

  8. 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

  9. Clinical and comparative utility of afatinib in non-small cell lung cancer

    PubMed Central

    D’Arcangelo, Manolo; Hirsch, Fred R

    2014-01-01

    The first targeted agents approved for non-small cell lung cancer (NSCLC) treatment, the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) gefitinib and erlotinib, have an impressive activity in the presence of activating mutations of the EGFR gene. However, all patients develop acquired resistance principally through secondary mutations (T790M), HER2 amplification, MET amplification, and other molecular aberrations. An attempt to overcome EGFR TKI resistance has been through the development of irreversible blockers. Afatinib is an irreversible inhibitor of the tyrosine kinase activity of all members of the HER family. The pharmacologic properties of afatinib (formation of covalent bonds, inhibition of other family members, and in vitro and in vivo activity on T790M mutation positive tumors) made this drug particularly appealing to study in clinic. Therefore, an intense program of clinical research (LUX-Lung program) was started and clinical results have shown very encouraging activity profiles in patients harboring EGFR activating mutations and in those with acquired resistance to reversible TKIs. PMID:24790411

  10. 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.

  11. Translational Treatment Paradigm for Managing Non-Unions Secondary to Radiation Injury Utilizing Adipose Derived Stem Cells and Angiogenic Therapy

    PubMed Central

    Donneys, Alexis; Blough, Jordan T.; Nelson, Noah S.; Perosky, Joseph E.; Deshpande, Sagar S.; Kang, Stephen Y.; Felice, Peter A.; Figueredo, Christian; Peterson, Jonathan R.; Kozloff, Kenneth M.; Levi, Benjamin; Chepeha, Douglas B.; Buchman, Steven R.

    2015-01-01

    Background Bony non-unions arising in the aftermath of collateral radiation injury are commonly managed with vascularized free tissue transfers. Unfortunately, these procedures are invasive and fraught with attendant morbidities. This study investigates a novel, alternative treatment paradigm utilizing adipose derived stem cells (ASCs) combined with angiogenic deferoxamine (DFO) in the rat mandible. Methods Rats were exposed to a bioequivalent dose of radiation and mandibular osteotomy. Those exhibiting non-unions were subsequently treated with surgical debridement alone or debridement plus combination therapy. Radiographic and biomechanical outcomes were assessed after healing. Results Significant increases in biomechanical strength and radiographic metrics were observed in response to combination therapy (p<0.05). Importantly, combined therapy enabled a 65% reduction in persisting non-unions when compared to debridement alone. Conclusions We support the continued investigation of this promising combination therapy in its potential translation for the management of radiation-induced bony pathology. PMID:25917284

  12. Energy storage capability of the dye sensitized solar cells via utilization of highly porous carbon electrodes

    NASA Astrophysics Data System (ADS)

    Rahimi, Fatemeh; Takshi, Arash

    2016-09-01

    Dye sensitized solar cells (DSSCs) have shown promising results in the field of renewable energy owing to their low cost and portable features. In practical applications, their harvested energy could be stored in a supercapacitor once it exceeds the regular consumption. Various methods of manipulation of the active electrode have been examined to facilitate the energy storage of the system, whereas the counter electrode has always been known for its catalytic functionality and its contribution to the capacitive response of the device left a well-oriented study to be desired. In this work, the substitution of the platinum electrode with a specific porous electrode resulted in a supercapacitive behavior of the device. The photoactive electrode was fabricated using zinc oxide nanowires (ZnO) grown on a conductive transparent substrate with hydrothermal deposition method. The electrode was used to make a standard DSSC using a ruthenium dye, iodide/triiodide standard redox electrolyte, and a platinum counter electrode. The cyclic voltammetry (CV) study of the device showed a low capacitance with 350 mV open circuit voltage. Replacing the platinum counter electrode with a particularly designed porous paper-based carbon nanotube electrode resulted in a considerable difference in the CV response. A capacitive behavior was observed due to the large surface area of the counter electrode and the ZnO nanostructures on the photoactive electrode. Due to the large capacitance and relatively small photocurrent, the change in the open circuit voltage was limited. However, enhancement of the photocurrent could improve both the energy harvesting and charge storage in the device.

  13. Differential pharmacology and clinical utility of sonidegib in advanced basal cell carcinoma

    PubMed Central

    Wahid, Mohd; Jawed, Arshad; Dar, Sajad Ahmad; Mandal, Raju K; Haque, Shafiul

    2017-01-01

    Patients suffering from advanced basal cell carcinoma (BCC) have very limited treatment options. Sonidegib selectively inhibits the growth of Hedgehog pathway-dependent tumors and can treat locally advanced BCC patients who are not candidates for surgery or radiation therapy. The BOLT clinical trials were conducted to evaluate the efficacy/potency of sonidegib in the treatment of advanced BCC or metastatic BCC. The patients were randomized in 1:2 ratios to receive 200 or 800 mg oral sonidegib daily, stratified by disease, histological subtype and geographical region. The primary efficacy analyses showed that 18 patients in the 200 mg group and 35 patients in the 800 mg group show an objective response (Central Review Committee) that corresponds to 43% (95% confidence interval [CI]: 28–59) and 38% (95% CI: 28–48) in their respective categories. Disease control was found in 93% (39 patients) and 80% (74 patients) of the patients administered 200 and 800 mg sonidegib, respectively. The adverse events were assessed by the Central Review Committee as well as the investigator review team as per the guidelines of National Cancer Institute Common Terminology Criteria for Adverse Events version 4.03. The most frequently found adverse events reported in BOLT trials were muscle spasms, alopecia, dysgeusia (taste disturbance), nausea, elevated blood creatine kinase and fatigue. Comparatively, the patients administered 200 mg sonidegib showed fewer adverse events than those in the 800 mg sonidegib category. Thus, the benefit of using the 200 mg dose of sonidegib outweighs the associated risks and it can be inferred that it would be judicious to choose doses of lesser strength. PMID:28182134

  14. Immunocytochemistry for amoxicillin and its use for studying uptake of the drug in the intestine, liver, and kidney of rats.

    PubMed

    Fujiwara, Kunio; Shin, Masashi; Miyazaki, Tsubasa; Maruta, Yasuhiro

    2011-01-01

    Specific transport systems for penicillins have been recognized, but their in vivo role in the context of other transporters remains unclear. We produced a serum against amoxicillin (anti-AMPC) conjugated to albumin with glutaraldehyde. The antiserum was specific for AMPC and ampicillin (ABPC) but cross-reacted weakly with cephalexin. This enabled us to develop an immunocytochemical (ICC) method for detecting the uptake of AMPC in the rat intestine, liver, and kidney. Three hours after a single oral administration of AMPC, the ICC method revealed that AMPC distributed to a high degree in the microvilli, nuclei, and cytoplasm of the absorptive epithelial cells of the intestine. AMPC distributed in the cytoplasm and nuclei of the hepatocytes in a characteristic granular morphology on the bile capillaries, and in addition, AMPC adsorption was observed on the luminal surface of the capillaries, intercalated portions, and interlobular bile ducts on the bile flow. Almost no AMPC could be detected 6 h postadministration in either the intestine or the liver. Meanwhile, in the kidney, AMPC persisted until 12 h postadministration to a high degree in the proximal tubules, especially in the S3 segment cells in the tubular lumen, in which numerous small bodies that strongly reacted with the antibody were observed. All these sites of AMPC accumulation correspond well to specific sites where certain transporter systems for penicillins occur, suggesting that AMPC is actually and actively absorbed, eliminated, or excreted at these sites, possibly through such certain penicillin transporters.

  15. CINtec PLUS immunocytochemistry as a tool for the cytologic diagnosis of glandular lesions of the cervix uteri.

    PubMed

    Ravarino, Alberto; Nemolato, Sonia; Macciocu, Elena; Fraschini, Matteo; Senes, Giancarlo; Faa, Gavino; Negri, Giovanni

    2012-11-01

    Cytologic findings of glandular lesions of the cervix uteri are often difficult to evaluate. We studied the usefulness of CINtec PLUS p16/Ki-67 double stain (mtm laboratories, Heidelberg, Germany) for the diagnosis of glandular lesions. The study included 47 abnormal results on liquid-based cytologic tests with a subsequent histologic diagnosis of adenocarcinoma in situ or with early invasion, and 16 samples with negative results on follow-up. All samples were stained with CINtec PLUS p16/Ki-67 double stain. Of the neoplastic samples, 7 were excluded because of insufficient residual cellularity or loss of neoplastic cells. Of the samples that were adequate, 92.5% were stained with CINtec PLUS, whereas 7.5% were judged inconclusive. All inconclusive cases were at least 3 years old. Of the 16 negative samples, 15 (93.8%) stained negative and only 1 (6.2%) showed several positive clusters of cells. Our study shows that CINtec PLUS is a robust and useful tool for the diagnosis of glandular lesions of the cervix uteri.

  16. [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.

  17. 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

  18. Optimization and utilization of the SureFire phospho-STAT5 assay for a cell-based screening campaign.

    PubMed

    Binder, Christina; Lafayette, Amy; Archibeque, Ivonne; Sun, Yu; Plewa, Cherylene; Sinclair, Angus; Emkey, Renee

    2008-02-01

    The family of signal transducers and activators of transcription (STATs) consists of seven transcription factors that respond to a variety of cytokines, hormones, and growth factors. STATs are activated by tyrosine phosphorylation, which results in their dimerization and translocation into the nucleus where they exert their effect on transcription of regulated target genes. The phosphorylation of STATs is mediated mainly by Janus kinases (JAKs). The JAK/STAT pathway plays a critical role in hematopoietic and immune cell function. Here we focus on one member of the STAT family, STAT5. STAT5 is phosphorylated by several JAKs, including Jak3, Jak2, and Tyk2, in response to interleukin-2, erythropoietin (EPO), and interleukin-22, respectively. Activation of STAT5 is essential to T cell development and has been associated with hematologic malignancies. Therefore, the ability to assess STAT5 phosphorylation is important for discovery efforts targeting these indications. The assay formats available to detect phosphorylated STAT5 (pSTAT5) are relatively low throughput and involve lengthy protocols. These formats include western blot analysis, enzyme-linked immunosorbent assay (ELISA), and flow cytometry. The SureFire (Perkin Elmer, Waltham, MA) pSTAT5 assay is a homogeneous assay that utilizes AlphaScreen (Perkin Elmer) technology to detect pSTAT5 in cell lysates. We have used this assay format to evaluate EPO-induced STAT5 phosphorylation in HEL cells and successfully complete a small-scale screening campaign to identify inhibitors of this event. The results obtained in these studies demonstrate that the SureFire pSTAT5 assay is a robust, reliable assay format that is amenable to high-throughput screening (HTS) applications.

  19. Platelet and Red Blood Cell Utilization and Transfusion Independence in Umbilical Cord Blood and Allogeneic Peripheral Blood Hematopoietic Cell Transplants

    PubMed Central

    Solh, Melhem; Brunstein, Claudio; Morgan, Shanna; Weisdorf, Daniel

    2010-01-01

    Allogeneic hematopoietic cell transplantation (HCT) recipients have substantial transfusion requirements. Factors associated with increased transfusions and the extent of blood product use in umbilical cord blood (UCB) recipients are uncertain. We reviewed blood product use in 229 consecutive adult recipients of allogeneic HCT at the University of Minnesota: 147 with leukemia, 82 lymphoma or myeloma; 58% received unrelated UCB and 43% sibling donor peripheral blood stem cell (PBSC) grafts. Although neutrophil recovery was prompt (UCB median 17, range 2–45 days, and PBSC 14, range 3–34 days), only 135 of 229 (59% cumulative incidence, CI) achieved RBC independence and 157 (69%) achieved platelet independence by 6 months. Time to platelet independence was prolonged in UCB recipients (median UCB 41 vs. PBSC 14 days) and in patients who had received a prior transplant (median 48 vs. 32 days). Patients who received UCB grafts required more RBC through day 60 post HCT (mean UCB 7.8 (95% CI 6.7–8.9) vs. PBSC 5.2 (3.7–6.7) transfusions, p=0.04), and more platelet transfusions (mean 25.2 (95% CI 22.1–28.2) vs. 12.9 (9.4–16.4), p<0.01) compared to PBSC recipients. Patient receiving myeloablative (MA) conditioning required more RBC and platelet transfusions during the first 2 months post HCT compared to reduced intensity conditioning (RIC) (7.4 vs. 6.2, p=0.3 for RBC; 23.2 vs 17.5, p=0.07 for platelets). Despite prompt neutrophil engraftment, UCB recipients had delayed platelet recovery as well as more prolonged and costly blood product requirements. Enhanced approaches to accelerate multilineage engraftment could limit the transfusion-associated morbidity and costs accompanying UCB allotransplantation. PMID:20813199

  20. 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.

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

    DOE PAGES

    Petrik, Deborah L.; Cass, Cynthia L.; Padmakshan, Dharshana; ...

    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

  2. Localization of a molluscan gonadotropin-releasing hormone in Aplysia californica by in situ hybridization and immunocytochemistry.

    PubMed

    Jung, Lisa H; Kavanaugh, Scott I; Sun, Biao; Tsai, Pei-San

    2014-01-01

    Gonadotropin-releasing hormone (GnRH) plays important roles in vertebrate reproduction. Recently, molecules structurally similar to vertebrate GnRH were discovered in mollusks, including a gastropod, Aplysia californica. As an important step toward understanding the function of A. californica GnRH (ap-GnRH), the present study examined the localization of ap-GnRH peptide and transcript in the central and peripheral tissues. Reverse transcription polymerase chain reaction (RT-PCR) revealed wide expression of ap-GnRH in all ganglia (abdominal, buccal, cerebral, and pedal ganglia) of the central nervous system (CNS) and in multiple peripheral organs. However, in situ hybridization (ISH) revealed that cells positive for ap-GnRH are detectable only in the CNS, with the pedal ganglia containing the highest number of ap-GnRH-positive neurons, followed by the cerebral and abdominal ganglia. Most neurons positive for the transcript were simultaneously positive for the peptide, although some discrepancies were observed in cerebral and abdominal ganglia. Overall, our data suggest the de novo synthesis of ap-GnRH is restricted to the CNS, with the pedal ganglia being the primary source of ap-GnRH. Our results support the notion that ap-GnRH is a bona-fide neuropeptide that may assume diverse central functions, including those unrelated to reproduction.

  3. An in vivo role of Mrp2 in the rat hepatocytes by immunocytochemistry for amoxicillin using the transporter-deficient EHBR.

    PubMed

    Fujiwara, Kunio; Shin, Masashi; Yoshizaki, Yohei; Miyazaki, Tsubasa; Saita, Tetsuya

    2012-06-01

    An in vivo role of the multidrug resistant-associated protein (Mrp2) in rat hepatocytes was examined by immunocytochemistry (ICC) for amoxicillin (AMPC) by the use of the transporter-deficient Eisai hyperbilirubinemic rats (EHBR). The ICC revealed that in the liver of EHBR at 3-h post-administration, amoxicillin accumulated in the cytoplasmic pools and nuclei of the hepatocytes in a characteristic granular morphology on the bile capillaries. However, no amoxicillin was observed on the surface of the lumina ranging from the bile capillaries to the interlobular bile ducts. The drug persisted at least for 6-h after administration. In contrast, in the control rat liver at 3-h post-administration, AMPC-adsorption occurred on such luminal surface, while AMPC accumulated to a less level in both the cytoplasm and nuclei of the hepatocytes. The drug completely disappeared in the hepatocytes at 6-h post-administration. These results strongly suggest that AMPC taken up into the cytoplasm of the hepatocytes excretes via Mrp2 into the bile flow. Furthermore, electron microscopy demonstrated that the lower electron density areas in large sizes, corresponding to the cytoplasmic pools in ICC for AMPC, occurred in the cytoplasm peripheral to the nuclei of the hepatocytes in EHBR at 3-h post-administration, and then disappeared 24 h after administration.

  4. Utilization of Mytilus digestive gland cells for the in vitro screening of potential metabolic disruptors in aquatic invertebrates.

    PubMed

    Balbi, Teresa; Ciacci, Caterina; Grasselli, Elena; Smerilli, Arianna; Voci, Adriana; Canesi, Laura

    2017-01-01

    In vertebrate systems, many endocrine disruptors (EDs) can also interfere with energy and lipid metabolism, thus acting as metabolic disruptors. At the cellular level, these effects are mainly mediated by interactions with nuclear receptors/transcription factors, leading to the modulation of genes involved in lipid homeostasis, as well as by rapid, receptor-independent pathways. Several potential metabolic disruptors are found in aquatic environments. In fish, different EDs have been shown to affect hepatic lipid homeostasis both in vivo and in vitro. However, little information is available in aquatic invertebrates due to our poor knowledge of the regulatory pathways of lipid metabolism. In this work, primary cell cultures from the digestive gland of the bivalve Mytilus galloprovincialis were utilized to investigate the effects of model EDs (bisphenol A (BPA) and perfluorooctane sulphonate (PFOS)) on lipid homeostasis. Both compounds (at 24 and 3h of exposure) increased intracellular lipid and tryglyceride-TAG content, with strongest effects of PFOS at 10(-7)M. Acyl-CoA oxidase activity was unaffected, whereas some changes in the activity of glycolytic, antioxidant/biotransformation enzymes were observed; however, no clear relationship was found with lipid accumulation. Evaluation of mitochondrial membrane potential Δψm and determination of extracellular TAG content indicate that PFOS interferes with mitochondrial function and lipid secretion, whereas BPA mainly affects lipid secretion. Experiments with specific inhibitors showed that activation of PI-3 kinase and extracellularly regulated mitogen-activated protein kinase (ERK MAPK) plays a key role in mediating lipid accumulation. Mussel digestive gland cells represent a simple in vitro model for screening the metabolic effects of EDs in marine invertebrates.

  5. Short-term regulation of muscarinic acetylcholine receptors: An assessment utilizing mouse brain and mouse neuroblastoma cells

    SciTech Connect

    Cioffi, C.L.

    1988-01-01

    The effects of muscarinic agonists and diisopropylfluorophosphate (DFP) on muscarinic receptor density and muscarinic receptor-mediated responses was assessed in mouse brain and mouse neuroblastoma cells (clone N1E-115). Utilizing the antagonist ({sup 3}H)quinuclidinyl benzilate (({sup 3}H)QNB), there was no difference in the maximal binding capacity (B{sub max}) or equilibrium dissociation constant (K{sub d}) between untreated and 24 hour DFP-treated mice. However, one administration of DFP produced a 24% and 33% decrease in B{sub max} measured by ({sup 3}H)N-methylscopolamine (({sup 3}H)NMS) after 18 and 24 hours which was rapidly reversible within 36 hours after DFP treatment. The loss of ({sup 3}H)NMS binding sites following acute DFP treatment was not accompanied by a change in a particular muscarinic receptor binding conformation. Furthermore, the magnitude of muscarinic receptor-mediated phosphoinositide hydrolysis was unchanged following short-term DFP treatment.

  6. 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.

  7. Effect of the state of distribution of supported Pt nanoparticles on effective Pt utilization in polymer electrolyte fuel cells.

    PubMed

    Uchida, Makoto; Park, Young-Chul; Kakinuma, Katsuyoshi; Yano, Hiroshi; Tryk, Donald A; Kamino, Takeo; Uchida, Hiroyuki; Watanabe, Masahiro

    2013-07-21

    In polymer electrolyte fuel cells, it is essential to minimize Pt loading, particularly at the cathode, without serious loss of performance. From this point of view, we will report an advanced concept for the design of high performance catalysts and membrane-electrode assemblies (MEAs): first, the evaluation of Pt particle distributions on both the interior and exterior walls of various types of carbon black (CB) particles used as supports with respect to the "effective surface (ES)"; second, control of both size and location of Pt particles by means of a new preparation method (nanocapsule method); and finally, a new evaluation method for the properties of MEAs based on the Pt utilization (UPt), mass activity (MA), and effectiveness of Pt (EfPt), based on the ES concept. The amounts of Pt catalyst particles located in the CB nanopores were directly evaluated using the transmission electron microscopy, scanning electron microscopy and corresponding three-dimensional images. By use of the nanocapsule method and optimization of the ionomer, increased MA and EfPt values for the MEA were achieved. The improvement in the cathode performance can be attributed to the sharp particle-size distribution for Pt and the highly uniform dispersion on the exterior surface of graphitized carbon black (GCB) supports.

  8. Design and optimization of anode flow field of a large proton exchange membrane fuel cell for high hydrogen utilization

    NASA Astrophysics Data System (ADS)

    Yesilyurt, Serhat; Rizwandi, Omid

    2016-11-01

    We developed a CFD model of the anode flow field of a large proton exchange membrane fuel cell that operates under the ultra-low stoichiometric (ULS) flow conditions which intend to improve the disadvantages of the dead-ended operation such as severe voltage transient and carbon corrosion. Very small exit velocity must be high enough to remove accumulated nitrogen, and must be low enough to retain hydrogen in the active area. Stokes equations are used to model the flow distribution in the flow field, Maxwell-Stefan equations are used to model the transport of the species, and a voltage model is developed to model the reactions kinetics. Uniformity of the distribution of hydrogen concentration is quantified as the normalized area of the region in which the hydrogen mole fraction remains above a certain level, such as 0.9. Geometry of the anode flow field is modified to obtain optimal configuration; the number of baffles at the inlet, width of the gaps between baffles, width of the side gaps, and length of the central baffle are used as design variables. In the final design, the hydrogen-depleted region is less than 0.2% and the hydrogen utilization is above 99%. This work was supported by The Scientific and Technolo-gical Research Council of Turkey, TUBITAK-213M023.

  9. Pharmacokinetics and red cell utilization of iron(III) hydroxide-sucrose complex in anaemic patients: a study using positron emission tomography.

    PubMed

    Beshara, S; Lundqvist, H; Sundin, J; Lubberink, M; Tolmachev, V; Valind, S; Antoni, G; Långström, B; Danielson, B G

    1999-02-01

    The pharmacokinetics of a single intravenous injection of 100 mg iron hydroxide-sucrose complex labelled with a tracer in the form of 52Fe/59Fe was followed in six anaemic patients for a period ranging from 6 to 8 3 h using positron emission tomography (PET). Red cell utilization of the labelled iron was followed for 4 weeks. PET data showed radioactive uptake by the liver, spleen and bone marrow. The uptake by the macrophage-rich spleen demonstrated the reticuloendothelial uptake of this iron preparation, with subsequent effective release of that iron for marrow utilization. Red cell utilization, followed for 4 weeks, ranged from 59% to 97%. The bone marrow influx rate constant was independent of blood iron concentration, indicating non-saturation of the transport system in bone marrow. This implied that higher doses of the iron complex can probably be used in the same setting. A higher influx rate into the marrow compared with the liver seemed to be consistent with higher red cell utilization. This would indicate that early distribution of the injected iron complex may predict the long-term utilization.

  10. The additional utility of apparent diffusion coefficient values of clear-cell renal cell carcinoma for predicting metastasis during clinical staging

    PubMed Central

    Yoshizako, Takeshi; Hisatoshi, Araki; Mori, Hiroshi; Tamaki, Yukihisa; Ishikawa, Noriyoshi; Kitagaki, Hajime

    2017-01-01

    Background The apparent diffusion coefficient (ADC) value is known to be an indicator of tumor activity. The ADC value of high-grade clear-cell renal cell carcinoma (RCC) is significantly lower than that of low-grade clear-cell RCC. Purpose To investigate the utility of ADC values of clear-cell RCC by comparing ADC values between groups with T1a RCC (tumor size ≤ 4 cm) without metastasis and the group with metastasis. Material and Methods A retrospective review was performed on 51 patients with 51 RCCs who underwent 1.5 T magnetic resonance imaging (MRI) for evaluating a renal mass confirmed pathologically to be clear-cell RCC between January 2010 and August 2014. We compared ADC values between group A (T1a RCC without metastasis, T1aN0M0) and group B (RCC with metastasis) using the Mann–Whitney U test. Results The patients were divided into group A (n = 30; tumor size: median, 24.5 mm; range, 8–40 mm; ADC value [×10−3 mm2/s]: median, 1.71; range, 1.23–2.24) and group B (n = 21; tumor size: median, 87.5 mm; range, 18–150 mm; ADC value [×10−3 mm2/s]: median, 1.35; range, 0.91–1.94). The ADC value differed significantly between the two groups. The area under the receiver operating characteristic curve was 0.869. Using the optimum cutoff value (1.552 × 10−3 mm2/s), ADC had a sensitivity of 80.0% and specificity of 81.0%. Conclusion There was a statistically significant difference in the ADC between group A (T1a clear-cell RCC without distant metastasis) and group B (advanced clear-cell RCC with lymph node metastasis or distant metastasis). PMID:28210496

  11. Natural OX40L expressed on human T cell leukemia virus type-I-immortalized T cell lines interferes with infection of activated peripheral blood mononuclear cells by CCR5-utilizing human immunodeficiency virus

    PubMed Central

    2013-01-01

    Background OX40 ligand (OX40L) co-stimulates and differentiates T cells via ligation of OX40 that is transiently induced on T cells upon activation, resulting in prolonged T cell survival and enhanced cytokine production by T cells. This view has led to the targeting of OX40 as a strategy to boost antigen specific T cells in the context of vaccination. In addition, the ligation of OX40 has also been shown to inhibit infection by CCR5-utilizing (R5) but not CXCR4-utilizing (X4) human immunodeficiency virus type-1 (HIV-1) via enhancement of production of CCR5-binding β-chemokines. It was reasoned that human T cell leukemia virus type-I (HTLV-1) immortalized T cell lines that express high levels of OX40L could serve as an unique source of physiologically functional OX40L. The fact that HTLV-1+ T cell lines simultaneously also express high levels of OX40 suggested a potential limitation. Results Results of our studies showed that HTLV-1+ T cell lines bound exogenous OX40 but not OX40L, indicating that HTLV-1+ T cell lines express an active form of OX40L but an inactive form of OX40. Anti-OX40 non-blocking monoclonal antibody (mAb), but not blocking mAb, stained HTLV-1+ T cell lines, suggesting that the OX40 might be saturated with endogenous OX40L. Functionality of the OX40L was confirmed by the fact that a paraformaldehyde (PFA)-fixed HTLV-1+ T cell lines inhibited the infection of autologous activated peripheral blood mononuclear cells (PBMCs) with R5 HIV-1 which was reversed by either anti-OX40L blocking mAb or a mixture of neutralizing mAbs against CCR5-binding β-chemokines. Conclusions Altogether, these results demonstrated that autologous T cell lines immortalized by HTLV-1 can be utilized as a conventional source of physiologically functional OX40L. PMID:24238037

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

    PubMed

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

    2016-02-02

    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.

  13. Labview utilities

    SciTech Connect

    Persaud, Arun

    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.

  14. Functional Characterization of Sodium-dependent Multivitamin Transporter (SMVT) in MDCK-MDR1 cells and its Utilization as a Target for Drug Delivery

    PubMed Central

    Luo, Shuanghui; Kansara, Viral S.; Zhu, Xiaodong; Pal, Dhananjay; Mitra, Ashim. K.

    2008-01-01

    The objective of this research is to characterize a sodium-dependent multivitamin transporter (SMVT) in MDCK-MDR1 cells (Madin-Darby canine kidney cells transfected with the human MDR1 gene) and to investigate the feasibility of utilizing MDCK-MDR1 cell line as an in vitro model to study the permeability of biotin-conjugated prodrugs of anti-HIV protease inhibitors. Mechanism of [3H] biotin uptake and transport was delineated. Transepithelial permeability of the biotin conjugated prodrug i.e. biotin-saquinavir was also studied. Reverse transcription-polymerase chain reaction (RT-PCR) was carried out to confirm the existence of SMVT in MDCK-MDR1 cells. Biotin uptake was Na+, pH, and temperature dependent, but energyindependent. Transepithelial transport studies of biotin-saquinavir in MDCK-MDR1, wild type MDCK, and Caco-2 cells revealed that permeability of biotin-saquinavir was similar in all three cell lines. A band of SMVT mRNA at 862 bp was identified by RT-PCR. A sodium-dependent multivitamin transporter, SMVT, responsible for biotin uptake and transport, was identified and functionally characterized in MDCK-MDR1 cells. Therefore, MDCK-MDR1 cell line may be utilized as an in vitro model to study the permeability of biotin conjugated prodrugs such as HIV protease inhibitors. PMID:16749865

  15. 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

  16. 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.

  17. Enhanced hydrogen production from waste activated sludge by cascade utilization of organic matter in microbial electrolysis cells.

    PubMed

    Lu, Lu; Xing, Defeng; Liu, Bingfeng; Ren, Nanqi

    2012-03-15

    Fermentative hydrogen production from waste activated sludge (WAS) has low H2 yield because WAS contains limited amounts of carbohydrate suitable for use by hydrogen-producing bacteria. Here, augmentation of hydrogen production from WAS by microbial electrolysis cells (MECs) was implemented. H2 yields of 3.89±0.39 mg-H2/g-DS (5.67±0.61 mg-H2/g-VSS) from raw WAS and 6.78±0.94 mg-H2/g-DS (15.08±1.41 mg-H2/g-VSS) from alkaline-pretreated WAS were obtained in the two-chamber MECs (TMECs). This was several times higher than yields obtained previously by fermentation. Single-chamber MECs (SMECs) with low internal resistance showed a H2 production rate that 13 times that of TMECs with similar H2 yield when alkaline-pretreated WAS was used. However, methanogenesis was detected after several batch cycles. A yield balance calculation revealed that carbohydrates were not the only substrates for electrohydrogenesis. Protein and its acidification products, such as volatile fatty acids are also responsible for a portion of H2 generation in MEC. Characterization of WAS in TMECs by three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy with parallel factor analysis indicated that electrohydrogenesis reacted on the extracellular polymeric substances and intracellular substances of WAS. Cascade utilization of organic matter in MECs increased hydrogen production from WAS. MECs showed high hydrogen yield from WAS, fewer H2 sinks, and insensitivity to temperature. Optimizing MEC configurations and operation conditions and improving the pretreatment processes of WAS are necessary before practical application can take place on a large scale.

  18. 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

    2014-07-09

    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.

  19. 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

  20. 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

  1. Sertoli cell condition medium can induce germ like cells from bone marrow derived mesenchymal stem cells

    PubMed Central

    Monfared, Mahdieh Hajian; Minaee, Bagher; Rastegar, Tayebeh; Khrazinejad, Ebrahim; Barbarestani, Mohammad

    2016-01-01

    Objective(s): Although many researchers have confirmed induction of germ cells from bone marrow mesenchymal stem cells (BMMSCs), there are no reports that confirm spontaneous differentiation of germ cells from BMMSCs. In this study, we have evaluated the effect of adult Sertoli cell condition medium (SCCM) as a mutative factor in the induction of germ cells from BMMSCs. Materials and Methods: BMMSCs were collected from the bone marrow of 6-8-week old NMRI mice and their mesenchymal entities were proven using superficial markers (expression of CD44 and CD73 and non-expresion of CD45 and CD11b) by fow cytometry. Their multi-potential entities were proved with differentiation to osteogenic and adipogenic cells for 21 days. Also isolated Sertoli cells were enriched using lectin coated plates and Sertoli cell condition medium (SCCM) was collected. Sertoli cells were identified by immunocytochemistry and Vimentin marker. The cells were then differentiated into germ cells with SCCM for 2 weeks. Finally induced cells were evaluated by RT-PCR and immunocytochemistry. Results: Differentiation of mesenchymal stem cells to osteoblast and adipocyte showed their multi-potential property. Expression of CD44 and CD73 and non-expression of CD45 and CD11b confirmed mesenchyme cells. Immunocytochemistry and RT-PCR results showed expression of germ cells specific marker (Mvh). Conclusion: This study confirmed the effect of SCCM as a motivational factor that can used for differentiation of germ cells from BMMSCs. PMID:27917274

  2. Multiple pass gas absorption cell utilizing a spherical mirror opposite one or more pair of obliquely disposed flat mirrors

    NASA Technical Reports Server (NTRS)

    Pearson, Richard (Inventor); Lynch, Dana H. (Inventor); Gunter, William D. (Inventor)

    1995-01-01

    A method and apparatus for passing light bundles through a multiple pass sampling cell is disclosed. The multiple pass sampling cell includes a sampling chamber having first and second ends positioned along a longitudinal axis of the sampling cell. The sampling cell further includes an entrance opening, located adjacent the first end of the sampling cell at a first azimuthal angular position. The entrance opening permits a light bundle to pass into the sampling cell. The sampling cell also includes an exit opening at a second azimuthal angular position. The light exit permits a light bundle to pass out of the sampling cell after the light bundle has followed a predetermined path.

  3. Clinical Utility of Serum Homocysteine and Folate as Tumor Markers in Oral Squamous Cell Carcinoma - A Cross-Sectional Study

    PubMed Central

    Erugula, Sridhar Reddy; Danappanavar, Prasanna M.; Ealla, Kranti Kiran Reddy; Velidandla, Surekha; Manikya, Sangameshwar

    2016-01-01

    Introduction Oral Squamous Cell Carcinoma (OSCC) is a common malignancy involving head and neck. Identifying the markers of molecular levels or biochemical markers involving the various metabolic reactions associated with the initiation and biological behavior of individual tumors are very important in diagnosis and prognosis. Aim To measure and compare the levels of serum Homocysteine (Hcy) and serum folate in OSCC patients, smoking group and healthy subjects and also to assess the clinical utility of serum Hcy as a potential tumor marker in OSCC. Materials and Methods The study group comprised of 60 subjects, of whom 30 were classified as OSCC cases (GROUP I) and 15 were classified as smokers without OSCC (GROUP II). The control group included 15 healthy individuals without smoking habit (Group III). Hcy was measured with High Performance Liquid Chromatography (HPLC). Folate estimation was done by Chemiluminiscence Immuno Assay (CLIA). Comparison of mean Hcy and folate values among the groups was done using ANOVA with Post-Hoc Games Howell test. Gender was compared using Chi-square test. Comparison of mean age was using ANOVA with Post-Hoc Tukey’s test. Results The mean serum folate level in OSCC patients was 5.34ng/mL, 7.68ng/mL in smoking group and 10.99ng/mL in control group. There was a significant difference in the mean serum folate levels among the three study groups (p<0.001). The mean serum Hcy in OSCC patients was 23.58μmol/L, 17.46μmol/L, in smoking group and 10.76μmol/l in controls. There was a significant difference in the mean serum Hcy levels among the three study groups (p<0.001). Conclusion The present study found an interesting association with serum Hcy and folate levels in OSCC which could be useful as a biochemical “Tumor Marker” and thereby providing insights into the onset and progression of the disease. PMID:27891452

  4. Prognostic utility of admission cell-free DNA levels in patients with chronic obstructive pulmonary disease exacerbations

    PubMed Central

    Avriel, Avital; Rozenberg, Dmitry; Raviv, Yael; Heimer, Dov; Bar-Shai, Amir; Gavish, Rachel; Sheynin, Jony; Douvdevani, Amos

    2016-01-01

    Background Chronic obstructive pulmonary disease exacerbations (COPDEs) are associated with increased morbidity and mortality. Cell-free DNA (cfDNA) is a novel biomarker associated with clinical outcomes in several disease states but has not been studied in COPD. The objectives of this study were to assess cfDNA levels during a COPDE, to evaluate the association of cfDNA with clinical parameters and to explore the prognostic implications of cfDNA levels on long-term survival. Methods This was an observational study that assessed cfDNA levels in patients admitted to hospital for a COPDE. Plasma cfDNA levels of COPDE patients were compared to those of matched stable COPD patients and healthy controls. Multivariable and Cox regression analyses were used to assess the association of cfDNA levels with blood gas parameters and long-term survival. Results A total of 62 patients (46 males, forced expiratory volume in 1 second [FEV1] 38%±13%) were included. The median cfDNA levels on admission for COPDE patients was 1,634 ng/mL (interquartile range [IQR] 1,016–2,319) compared to 781 ng/mL (IQR 523–855) for stable COPD patients, matched for age and disease severity, and 352 ng/mL (IQR 209–636) for healthy controls (P<0.0001, for both comparisons). cfDNA was correlated with partial arterial pressure of carbon dioxide (PaCO2, r=0.35) and pH (r=−0.35), P=0.01 for both comparisons. In a multivariable analysis, PaCO2 was the only independent predictor of cfDNA. Using a cfDNA level of 1,924 ng/mL (threshold for abnormal PaCO2), those with high levels had a trend for increased 5-year mortality risk adjusted for age, sex and FEV1% (hazard ratio 1.92, 95% confidence interval 0.93–3.95, P=0.08). Conclusion Plasma cfDNA might offer a novel technique to identify COPD patients at increased risk of poor outcomes, but the prognostic utility of this measurement requires further study. PMID:28003743

  5. 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...

  6. Construction of a xylan-fermenting yeast strain through codisplay of xylanolytic enzymes on the surface of xylose-utilizing Saccharomyces cerevisiae cells.

    PubMed

    Katahira, Satoshi; Fujita, Yasuya; Mizuike, Atsuko; Fukuda, Hideki; Kondo, Akihiko

    2004-09-01

    Hemicellulose is one of the major forms of biomass in lignocellulose, and its essential component is xylan. We used a cell surface engineering system based on alpha-agglutinin to construct a Saccharomyces cerevisiae yeast strain codisplaying two types of xylan-degrading enzymes, namely, xylanase II (XYNII) from Trichoderma reesei QM9414 and beta-xylosidase (XylA) from Aspergillus oryzae NiaD300, on the cell surface. In a high-performance liquid chromatography analysis, xylose was detected as the main product of the yeast strain codisplaying XYNII and XylA, while xylobiose and xylotriose were detected as the main products of a yeast strain displaying XYNII on the cell surface. These results indicate that xylan is sequentially hydrolyzed to xylose by the codisplayed XYNII and XylA. In a further step toward achieving the simultaneous saccharification and fermentation of xylan, a xylan-utilizing S. cerevisiae strain was constructed by codisplaying XYNII and XylA and introducing genes for xylose utilization, namely, those encoding xylose reductase and xylitol dehydrogenase from Pichia stipitis and xylulokinase from S. cerevisiae. After 62 h of fermentation, 7.1 g of ethanol per liter was directly produced from birchwood xylan, and the yield in terms of grams of ethanol per gram of carbohydrate consumed was 0.30 g/g. These results demonstrate that the direct conversion of xylan to ethanol is accomplished by the xylan-utilizing S. cerevisiae strain.

  7. Pharmacokinetics and red cell utilization of 52Fe/59Fe-labelled iron polymaltose in anaemic patients using positron emission tomography.

    PubMed

    Beshara, Soheir; Sörensen, Jens; Lubberink, Mark; Tolmachev, Vladimir; Långström, Bengt; Antoni, Gunnar; Danielson, Bo G; Lundqvist, Hans

    2003-03-01

    Parenteral iron-polysaccharide complexes are increasingly applied. The pharmacokinetics of iron sucrose have been assessed by our group using positron emission tomography (PET). A single intravenous injection of 100 mg iron as iron (III) hydroxide-polymaltose complex, labelled with a tracer in the form of 52Fe/59Fe, was similarly assessed in six patients using PET for about 8 h. Red cell utilization was followed for 4 weeks. Iron polymaltose was similarly distributed to the liver, spleen and bone marrow. However, a larger proportion of this complex was rapidly distributed to the bone marrow. The shorter equilibration phase for the liver, about 25 min, indicates the minimal role of the liver for direct distribution. Splenic uptake also reflected the reticuloendothelial handling of this complex. Red cell utilization ranged from 61% to 99%. Despite the relatively higher uptake by the bone marrow, there was no saturation of marrow transport systems at this dose level. In conclusion, high red cell utilization of iron polymaltose occurred in anaemic patients. The major portion of the injected dose was rapidly distributed to the bone marrow. In addition, the reticuloendothelial uptake of this complex may reflect the safety of polysaccharide complexes. Non-saturation of transport systems to the bone marrow indicated the presence of a large interstitial transport pool, which might possibly be transferrin.

  8. Utilization of pyrolytic substrate by microalga Chlamydomonas reinhardtii: cell membrane property change as a response of the substrate toxicity.

    PubMed

    Zhao, Xuefei; Jarboe, Laura; Wen, Zhiyou

    2016-05-01

    Acetic acid derived from fast pyrolysis of lignocellulosic biomass is a promising substrate for microalgae fermentation for producing lipid-rich biomass. However, crude pyrolytic acetic acid solution contains various toxic compounds inhibiting algal growth. It was hypothesized that such an inhibition was mainly due to the cell membrane damage. In this work, the cell membrane property of algal cells was evaluated at various conditions to elucidate the mechanisms of inhibition caused by the pyrolytic substrate solution. It was found that acetic acid itself served a carbon source for boosting algal cell growth but also caused cell membrane leakage. The acetic acid concentration for highest cell density was 4 g/L. Over-liming treatment of crude pyrolytic acetic acid increased the algal growth with a concurrent reduction of cell membrane leakage. Directed evolution of algal strain enhanced cell membrane integrity and thus increased its tolerance to the toxicity of the crude substrate. Statistical analysis shows that there was a significant correlation between the cell growth performance and the cell membrane integrity (leakage) but not membrane fluidity. The addition of cyto-protectants such as Pluronic F68 and Pluronic F127 enhanced the cell membrane integrity and thus, resulted in enhanced cell growth. The transmission electron microscopy (TEM) of algal cells visually confirmed the cell membrane damage as the mechanism of the pyrolytic substrate inhibition. Collectively, this work indicates that the cell membrane is one major reason for the toxicity of pyrolytic acetic acid when being used for algal culture. To better use this pyrolytic substrate, cell membrane of the microorganism needs to be strengthened through either strain improvement or addition of membrane protectant reagents.

  9. 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.

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

    SciTech Connect

    Walsh, Richard Joseph

    2002-01-01

    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 (fΔμ)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 Δμ 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 single cells is

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

    SciTech Connect

    Walsh, Richard Joseph

    2002-01-01

    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 (fΔμ) 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 Δμ 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 single cells is

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

    ClinicalTrials.gov

    2017-02-20

    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. 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.

  14. 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.

  15. 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

  16. Utility of Quantitative Flow Cytometry Immunophenotypic Analysis of CD5 Expression in Small B-cell Neoplasms

    PubMed Central

    Challagundla, Pramoda; Jorgensen, Jeffrey L.; Kanagal-Shamanna, Rashmi; Gurevich, Inga; Pierson, Diane M.; Ferrajoli, Alessandra; Reyes, Steven R.; Medeiros, L. Jeffrey; Miranda, Roberto N.

    2014-01-01

    Background The value of assessing CD5 expression in the differential diagnosis of small B-cell neoplasms is well established. Usually CD5 is assessed qualitatively. Materials and Methods We assessed CD5 expression levels by quantitative flow cytometry immunophenotyping to determine possible differences among various small B-cell neoplasms. We performed 4-color flow cytometry analysis on peripheral blood (PB) and bone marrow (BM) aspirate specimens and quantified CD5 expression in various mature small B-cell lymphomas and leukemias. We also assessed CD5 levels in PB samples of normal donors (controls). Results Cases of chronic lymphocytic leukemia (CLL) and mantle cell lymphoma had higher levels of CD5 compared with control (benign) B-cells (p < 0.001). Cases of marginal zone lymphoma (MZL) and hairy cell leukemia (HCL) had CD5 levels similar to control B-cells (p > 0.05), while cases of follicular lymphoma (FL) and lymphoplasmacytic lymphoma (LPL) had significantly lower CD5 levels than control B-cells (p <0.05). In B-cell neoplasms, a high level of CD5 expression correlated with a homogeneous pattern of positive events whereas lower CD5 levels correlated with a heterogeneous pattern of positive events. Conclusions Using flow cytometric immunophenotypic analysis to quantify CD5 levels can aid in diagnosis. CD5 expression levels are substantially higher in CLL and mantle cell lymphoma and expression is observed in a homogeneous pattern as compared with other B-cell neoplasms that are either negative for CD5 or express this antigen at lower levels with a heterogeneous pattern of expression. However, there is some overlap in CD5 expression levels between a subset of atypical CLL and MZL cases. PMID:24978916

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

    PubMed

    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.

  18. 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

  19. 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.

  20. Enhanced conversion of induced neuronal cells (iN cells) from human fibroblasts: Utility in uncovering cellular deficits in mental illness-associated chromosomal abnormalities.

    PubMed

    Passeri, Eleonora; Wilson, Ashley M; Primerano, Amedeo; Kondo, Mari A; Sengupta, Srona; Srivastava, Rupali; Koga, Minori; Obie, Cassandra; Zandi, Peter P; Goes, Fernando S; Valle, David; Rapoport, Judith L; Sawa, Akira; Kano, Shin-ichi; Ishizuka, Koko

    2015-12-01

    The novel technology of induced neuronal cells (iN cells) is promising for translational neuroscience, as it allows the conversion of human fibroblasts into cells with postmitotic neuronal traits. However, a major technical barrier is the low conversion rate. To overcome this problem, we optimized the conversion media. Using our improved formulation, we studied how major mental illness-associated chromosomal abnormalities may impact the characteristics of iN cells. We demonstrated that our new iN cell culture protocol enabled us to obtain more precise measurement of neuronal cellular phenotypes than previous iN cell methods. Thus, this iN cell culture provides a platform to efficiently obtain possible cellular phenotypes caused by genetic differences, which can be more thoroughly studied in research using other human cell models such as induced pluripotent stem cells.

  1. Regulation of the utilization of mRNA for eucaryotic elongation factor Tu in Friend erythroleukemia cells.

    PubMed Central

    Rao, T R; Slobin, L I

    1987-01-01

    When Friend erythroleukemia cells were allowed to grow to stationary phase (2 X 10(6) to 3 X 10(6) cells per ml), approximately 60% of the mRNA for eucaryotic elongation factor Tu (eEF-Tu) sedimented at less than or equal to 80S, and most of the remaining factor mRNA was associated with small polysomes. Under the same growth conditions, greater than 90% of the mRNA for eucaryotic initiation factor 4A remained associated with polysomes. The association of eEF-Tu mRNA with polysomes changed dramatically when stationary-phase cells were treated with fresh medium. After 1 h in fresh medium, approximately 90% of eEF-Tu mRNA in Friend cells was found in heavy polysomes. Associated with the shift of eEF-Tu mRNA into heavy polysomes, we found at least a 2.6-fold increase in the synthesis of eEF-Tu in vivo as well as a remarkable 40% decrease in the total amount of eEF-Tu mRNA per cell. Our data raise the possibility that eEF-Tu mRNA that has accumulated in ribonucleoprotein particles in stationary-phase cells is degraded rather than reutilized for eEF-Tu synthesis. Images PMID:2434834

  2. Shot number estimation for EB direct writing for logic LSI utilizing character-build standard-cell layout technique

    NASA Astrophysics Data System (ADS)

    Kajiya, Yoshihiko; Nakamura, Akihiro; Yoshikawa, Masaya; Fujino, Takeshi

    2006-05-01

    Electron Beam direct writing (EBDW) technology is the most cost-effective lithography tool for small-volume logic-LSI fabrication. The EB exposure time will be greatly reduced by applying character-projection (CP) aperture. But the applicable number of CP aperture is limited to 25-400 depending upon EB lithography apparatus. The cell-based logic LSIs are composed of standard-cells (SCs) whose number is 400-1000. Therefore, it is impossible to implement all SCs as CP apertures, because the SCs are placed to 4-directions in general. We had proposed the new technique named 'Character-Build (CB) standard-cell', and demonstrate the most of the combination-logic SCs can be composed by only 17 CP apertures. In this paper, not only combination-logic SCs but also sequential-logic SCs are considered. The number of EB-shots and the chip-area are estimated for some sample circuits. Compared to the simply-limited SCs, The EB shot number is 30-40% reduced by using proposed CB standard-cell, when the CP aperture numbers are 20-30. Moreover, CB standard-cell was advantageous in the module area. Considering 2-directional placement of SCs, the combination of the EB apparatus with 50-100 CP apertures and the CB standard-cell technique may be the best method for high-speed EB direct-writing.

  3. Phosphatidylserine biosynthesis in cultured Chinese hamster ovary cells. III. Genetic evidence for utilization of phosphatidylcholine and phosphatidylethanolamine as precursors

    SciTech Connect

    Kuge, O.; Nishijima, M.; Akamatsu, Y.

    1986-05-05

    We reported that Chinese hamster ovary (CHO) cells contain two different serine-exchange enzymes (I and II) which catalyze the base-exchange reaction of phospholipid(s) with serine and that a phosphatidylserine-requiring mutant (strain PSA-3) of CHO cells is defective in serine-exchange enzyme I and lacks the ability to synthesize phosphatidylserine. In this study, we examined precursor phospholipids for phosphatidylserine biosynthesis in CHO cells. When mutant PSA-3 and parent (CHO-K1) cells were cultured with (/sup 32/P)phosphatidylcholine, phosphatidylserine in the parent accumulated radioactivity while that in the mutant was not labeled significantly. On the contrary, when cultured with (/sup 32/P)phosphatidylethanolamine, the mutant incorporated the label into phosphatidylserine more efficiently than the parent. Furthermore, we found that mutant PSA-3 grew normally in growth medium supplemented with 30 microM phosphatidylethanolamine as well as phosphatidylserine and that the biosynthesis of phosphatidylserine in the mutant was normal when cells were cultured in the presence of exogenous phosphatidylethanolamine. The simplest interpretation of these findings is that phosphatidylserine in CHO cells is biosynthesized through the following sequential reactions: phosphatidylcholine----phosphatidylserine----phosphatidylethanolamine--- - phosphatidylserine. The three reactions are catalyzed by serine-exchange enzyme I, phosphatidylserine decarboxylase, and serine-exchange enzyme II, respectively.

  4. 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.

  5. Utilizing induced pluripotent stem cells (iPSCs) to understand the actions of estrogens in human neurons

    PubMed Central

    Shum, Carole; Macedo, Sara C.; Warre-Cornish, Katherine; Cocks, Graham; Price, Jack; Srivastava, Deepak P.

    2015-01-01

    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

  6. Normal and defective repair of damaged DNA in human cells: a sensitive assay utilizing the photolysis of bromodeoxyuridine.

    PubMed

    Regan, J D; Setlow, R B; Ley, R D

    1971-04-01

    A new technique has been developed for studying the extent of repair of UV-radiation damage to DNA in human cells. It is easy to use, has excellent sensitivity, and provides rapid quantitative estimates of repair. UV-irradiated cells whose DNA has been previously labeled with a radioisotope are grown after irradiation in non-radioactive bromodeoxyuridine, which is incorporated at the breaks induced by repair enzymes. After a period of growth in the thymidine analog the cells are exposed to a large flux of 313 nm radiation and then lysed on top of an alkaline sucrose gradient. Bromodeoxyuridine-containing sections of the DNA are thus selectively photolysed. Sedimentation in the alkaline gradient reveals the average molecular weight of disrupted segments and gives a measure of the number of breaks induced by repair enzymes over the whole period allowed for repair. The large change in average molecular weight observed upon exposure of normal repairing cells to 313 nm radiation is not observed in the repair-deficient cells from patients with xeroderma pigmentosum. The quantitative aspects of this assay for repair and its sensitivity should make it applicable to the study of repair induced by agents other than UV radiation.

  7. Utilization of Tabula Rasa to Stabilize Bulk Lifetimes in n-Cz Silicon for High-Performance Solar Cell Processing

    SciTech Connect

    LaSalvia, Vincenzo; Jensen, Mallory Ann; Youssef, Amanda; Nemeth, William; Page, Matthew; Buonassisi, Tonio; Stradins, Paul

    2016-11-21

    We investigate a high temperature, high cooling-rate anneal Tabula Rasa (TR) and report its implications on n-type Czochralski-grown silicon (n-Cz Si) for photovoltaic fabrication. Tabula Rasa aims at dissolving and homogenizing oxygen precipitate nuclei that can grow during the cell process steps and degrade the cell performance due to their high internal gettering and recombination activity. The Tabula Rasa thermal treatment is performed in a clean tube furnace with cooling rates >100 degrees C/s. We characterize the bulk lifetime by Sinton lifetime and photoluminescence mapping just after Tabula Rasa, and after the subsequent cell processing. After TR, the bulk lifetime surprisingly degrades to <; 0.1ms, only to recover to values equal or higher than the initial non-treated wafer (several ms), after typical high temperature cell process steps. Those include boron diffusion and oxidation; phosphorus diffusion/oxidation; ambient annealing at 850 degrees C; and crystallization annealing of tunneling-passivating contacts (doped polycrystalline silicon on 1.5 nm thermal oxide). The drastic lifetime improvement during high temperature cell processing is attributed to improved external gettering of metal impurities and annealing of intrinsic point defects. Time and injection dependent lifetime spectroscopy further reveals the mechanisms of lifetime improvement after Tabula Rasa treatment. Additionally, we report the efficacy of Tabula Rasa on n-type Cz-Si wafers and its dependence on oxygen concentration, correlated to position within the ingot.

  8. Hybrid organic-inorganic heterojunction solar cells with 12% efficiency by utilizing flexible film-silicon with a hierarchical surface.

    PubMed

    Thiyagu, Subramani; Hsueh, Chen-Chih; Liu, Chien-Ting; Syu, Hong-Jhang; Lin, Tzu-Ching; Lin, Ching-Fuh

    2014-03-21

    This paper reports an organic-inorganic hybrid solar cell with a hierarchical surface composed of high density silicon nanoholes and micro-desert textures. High-efficiency organic-inorganic hybrid solar cell Si/PEDOT-PSS with a hierarchical surface, showing a power conversion efficiency of 12%. The structure provides excellent light absorption over 97% for the spectral range of 300 to 1100 nm with a thickness of 60 μm due to internal multiple reflections caused by subwavelength features of high density silicon nanoholes and micro-desert textures. In addition, from the angle of incidence (AOI) observed, even at the large angle of 75°, the reflectance value still exhibits less than 1%. With the advantage of very thin silicon material and inexpensive processing, hybrid silicon/polymer solar cells are promising for various applications and thus could be an economically feasible alternative energy solution in the future.

  9. Hybrid organic-inorganic heterojunction solar cells with 12% efficiency by utilizing flexible film-silicon with a hierarchical surface

    NASA Astrophysics Data System (ADS)

    Thiyagu, Subramani; Hsueh, Chen-Chih; Liu, Chien-Ting; Syu, Hong-Jhang; Lin, Tzu-Ching; Lin, Ching-Fuh

    2014-02-01

    This paper reports an organic-inorganic hybrid solar cell with a hierarchical surface composed of high density silicon nanoholes and micro-desert textures. High-efficiency organic-inorganic hybrid solar cell Si/PEDOT-PSS with a hierarchical surface, showing a power conversion efficiency of 12%. The structure provides excellent light absorption over 97% for the spectral range of 300 to 1100 nm with a thickness of 60 μm due to internal multiple reflections caused by subwavelength features of high density silicon nanoholes and micro-desert textures. In addition, from the angle of incidence (AOI) observed, even at the large angle of 75°, the reflectance value still exhibits less than 1%. With the advantage of very thin silicon material and inexpensive processing, hybrid silicon/polymer solar cells are promising for various applications and thus could be an economically feasible alternative energy solution in the future.This paper reports an organic-inorganic hybrid solar cell with a hierarchical surface composed of high density silicon nanoholes and micro-desert textures. High-efficiency organic-inorganic hybrid solar cell Si/PEDOT-PSS with a hierarchical surface, showing a power conversion efficiency of 12%. The structure provides excellent light absorption over 97% for the spectral range of 300 to 1100 nm with a thickness of 60 μm due to internal multiple reflections caused by subwavelength features of high density silicon nanoholes and micro-desert textures. In addition, from the angle of incidence (AOI) observed, even at the large angle of 75°, the reflectance value still exhibits less than 1%. With the advantage of very thin silicon material and inexpensive processing, hybrid silicon/polymer solar cells are promising for various applications and thus could be an economically feasible alternative energy solution in the future. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr06323b

  10. The Utilization of HOPG based Graphene for a less costly and efficient replacement of platinum in Dye-Sensitized Solar Cells (DSC's)

    NASA Astrophysics Data System (ADS)

    Pusko, Matthew P.

    2012-10-01

    Dye-Sensitized Solar Cells (DSC) are also known as Graetzel cells after their inventor, Michael Graetzel, who invented them in 1991. DSCs are potential alternatives to the more conventional and expensive semiconductor p-n junction solar cells like silicon solar cells. In a Graetzel cell, light is absorbed by a sensitizing dye which is coupled to a wide band gap semiconductor (TiO2). Electric charges are liberated by the photon induced electron injection from the dye molecules into the conduction band of the semiconductor. The use of sensitizers in conjunction with the semiconductor oxide permits the absorption of a large portion of the terrestrial solar spectrum with near 10% conversion efficiency. In the ``traditional'' DSC, platinum metal is used as one of the counter electrode materials. In our research, graphene of various forms (from HOPG in specific) is utilized instead of costly platinum to replace the counter-electrode in the DSC's. HOPG also known as Higly Oriented Pyrolitic Graphite is being used to dry deposit graphene layers onto glass or FTO.

  11. Factors controlling oxygen utilization.

    PubMed

    Biaglow, John; Dewhirst, Mark; Leeper, Dennis; Burd, Randy; Tuttle, Steve

    2005-01-01

    We demonstrate, theoretically, that oxygen diffusion distance is related to the metabolic rate of tumors (QO2) as well as the oxygen tension. The difference in QO2 rate between tumors can vary by as much as 80-fold. Inhibition of oxygen utilization by glucose or chemical inhibitors can improve the diffusion distance. Combining respiratory inhibitors with increased availability of oxygen will further improve the oxygen diffusion distance for all tumors. A simple means for inhibiting oxygen consumption is the use of glucose (the Crabtree effect). The inhibition of tumor oxygen utilization by glucose occurs in R323OAc mammary carcinoma and 9L glioma cells. However, stimulation of oxygen consumption is observed with glucose in the Q7 hepatoma cell line. MIBG, a known inhibitor of oxygen utilization, blocks oxygen consumption in 9L, but is weakly inhibitory with the Q7. Q7 tumor cells demonstrate an anomalous behavior of glucose and MIBG on oxygen consumption. Our results clearly demonstrate the necessity for comparing effects of different agents on different tumor cells. Generalizations cannot be made with respect to the choice of inhibitor for in vivo use. Our work shows that oxygen consumption also can be inhibited with malonate and chlorosuccinate. These substrates may be effective in vivo, where glucose is low and glutamine is the major substrate. Our results indicate that information about individual tumor substrate-linked metabolic controls may be necessary before attempting to inhibit oxygen utilization in vivo for therapeutic benefit.

  12. Optimization of wastewater feeding for single-cell protein production in an anaerobic wastewater treatment process utilizing purple non-sulfur bacteria in mixed culture condition.

    PubMed

    Honda, Ryo; Fukushi, Kensuke; Yamamoto, Kazuo

    2006-10-01

    Impacts of operation timing of feeding and withdrawal on anaerobic wastewater treatment utilizing purple non-sulfur bacteria have been investigated in mixed culture condition with acidogenic bacteria. Simulated wastewater containing glucose was treated in a laboratory-scale chemostat reactor, changing the timing of wastewater feeding and withdrawal. Rhodopseudomonas palustris, which does not utilize glucose as a substrate, was inoculated in the reactor. Rps. palustris was detected by a fluorescent in situ hybridization (FISH) technique using the specific Rpal686 probe. As a result, population ratios of Rps. palustris were over 20% through the operation. Rps. palustris could grow by utilizing metabolites of acidogenic bacteria that coexisted in the reactor. A morning feed was effective for a good growth of purple non-sulfur bacteria. A protein content of cultured bacteria was the highest when wastewater was fed in the morning. Dissolved organic carbon (DOC) removal was 94% independent of the timing control. Consequently, feeding in the morning is the optimum feed-timing control from the aspects of growth of purple non-sulfur bacteria and single-cell protein production.

  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. 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.

  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. Eosinophil activation of fibroblasts from chronic allergen-induced disease utilizes stem cell factor for phenotypic changes.

    PubMed

    Dolgachev, Vladislav; Berlin, Aaron A; Lukacs, Nicholas W

    2008-01-01

    In the present studies the role of stem cell factor (SCF) in mediating eosinophil and fibroblast activation during their interaction was investigated. SCF was significantly higher in fibroblasts grown from lungs of chronic allergen-challenged mice compared to fibroblasts grown from normal mice. When eosinophils were layered onto fibroblasts from allergic mice, a significant increase in SCF was detected compared to fibroblasts from nonallergic mice. The interaction of fibroblasts with eosinophils also increased the production of asthma-associated chemokines, CCL5 and CCL6, was dependent on cell-to-cell interaction, and was observed only with fibroblasts derived from lungs of chronic allergen-challenged mice and not from those derived from unchallenged normal mice. Chemokine production was significantly decreased when anti-SCF antibodies were added during eosinophil-fibroblast interaction. The interaction of fibroblasts from chronic allergen-challenged mice with eosinophils also increased alpha-smooth muscle cell actin and procollagen I expression as well as induced transforming growth factor-beta. The changes in myofibroblast activation were dependent on SCF-mediated pathways because anti-SCF antibody treatment reduced the expression of all three of these latter fibrosis-associated markers. Thus, our data suggest that SCF mediates an important activation pathway for fibroblasts during chronic allergic responses on interaction with recruited eosinophils and suggest a potential mechanism of airway remodeling during chronic disease.

  18. 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.

  19. Handheld confocal laser endomicroscopic imaging utilizing tumor-specific fluorescent labeling to identify experimental glioma cells in vivo

    PubMed Central

    Martirosyan, Nikolay L.; Georges, Joseph; Kalani, M. Yashar S.; Nakaji, Peter; Spetzler, Robert F.; Feuerstein, Burt G.; Preul, Mark C.

    2016-01-01

    Background: We have reported that handheld confocal laser endomicroscopy (CLE) can be used with various nonspecific fluorescent dyes to improve the microscopic identification of brain tumor and its boundaries. Here, we show that CLE can be used experimentally with tumor-specific fluorescent labeling to define glioma margins in vivo. Methods: Thirteen rats underwent craniectomy and in vivo imaging 21 days after implantation with green fluorescent protein (GFP)-labeled U251 (n = 7) cells or epidermal growth factor receptor (EGFR) overexpressing F98 cells (n = 6). Fluorescein isothiocyanate (FITC) conjugated EGFR fluorescent antibody (FITC-EGFR) was applied for contrast in F98 tumors. Confocal images of normal brain, obvious tumor, and peritumoral zones were collected using the CLE system. Bench-top confocal microscopy and hematoxylin and eosin-stained sections were correlated with CLE images. Results: GFP and FITC-EGFR fluorescence of glioma cells were detected by in vivo visible-wavelength fluorescence CLE. CLE of GFP-labeled tumors revealed bright individual satellite tumor cells within peritumoral tissue, a definitive tumor border, and subcellular structures. Imaging with FITC-EGFR labeling provided weaker contrast in F98-EGFR tumors but was able to delineate tumor cells. Imaging with both methods in various tumor regions correlated with standard confocal imaging and clinical histology. Conclusions: These data suggest that in vivo CLE of selectively tagged neoplasms could allow specific interactive identification of tumoral areas. Imaging of GFP and FITC-EGFR provides real-time histologic information precisely related to the site of microscopic imaging of tumor. PMID:28144472

  20. 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.

  1. Utilization of TREC and KREC quantification for the monitoring of early T- and B-cell neogenesis in adult patients after allogeneic hematopoietic stem cell transplantation

    PubMed Central

    2013-01-01

    Background After hematopoietic stem cell transplantation (HSCT) T- and B-cell reconstitution from primary lymphoid organs are a prerequisite for an effective early lymphocyte reconstitution and a long-term survival for adult patients suffering from acute leukemia. Here, we asked whether quantification of T cell receptor excision circle, (TREC) and kappa-deleting recombination excision circle (KREC) before and within six month after allogeneic HSCT could be used to measure the thymic and bone marrow outputs in such patients. Methods We used a duplex real time PCR assay to quantify the absolute copy counts of TREC and KREC, and correlated the data with absolute cell counts of CD3+CD4+ T-cell and CD19+ B-cell subsets determined by flow cytometry, respectively. Results By comparing two recently proposed naïve T cell subsets, CD31+ naive and CD31- naive T cells, we found a better correlation for the CD31+ subset with TREC level post alloHSCT, in line with the assumption that it contained T cells recently derived from the thymus, indicating that TREC levels reflected real thymic de novo production. Transitional as well as naïve B cells highly correlated with KREC levels, which suggested an association of KREC levels with ongoing bone marrow B cell output. CD45RO+ memory T cells and CD27+ memory B cells were significantly less correlated with TREC and KREC recovery, respectively. Conclusion We conclude that simultaneous TREC/ KREC quantification is as a suitable and practicable method to monitor thymic and bone marrow output post alloHSCT in adult patients diagnosed with acute leukemia. PMID:23941115

  2. Diagnostic Utility of Pleural Fluid Cell Block versus Pleural Biopsy Collected by Flex-Rigid Pleuroscopy for Malignant Pleural Disease: A Single Center Retrospective Analysis

    PubMed Central

    Sasada, Shinji; Izumo, Takehiro; Matsumoto, Yuji; Tsuchida, Takaaki

    2016-01-01

    Background Some trials recently demonstrated the benefit of targeted treatment for malignant disease; therefore, adequate tissues are needed to detect the targeted gene. Pleural biopsy using flex-rigid pleuroscopy and pleural effusion cell block analysis are both useful for diagnosis of malignancy and obtaining adequate samples. The purpose of our study was to compare the diagnostic utility between the two methods among patients with malignant pleural disease with effusion. Methods Data from patients who underwent flex-rigid pleuroscopy for diagnosis of pleural effusion suspicious for malignancy at the National Cancer Center Hospital, Japan between April 2011 and June 2014 were retrospectively reviewed. All procedures were performed under local anesthesia. At least 150 mL of pleural fluid was collected by pleuroscopy, followed by pleural biopsies from the abnormal site. Results Thirty-five patients who were finally diagnosed as malignant pleural disease were included in this study. Final diagnoses of malignancy were 24 adenocarcinoma, 1 combined adeno-small cell carcinoma, and 7 malignant pleural mesothelioma (MPM), and 3 metastatic breast cancer. The diagnostic yield was significantly higher by pleural biopsy than by cell block [94.2% (33/35) vs. 71.4% (25/35); p = 0.008]. All patients with positive results on cell block also had positive results on pleural biopsy. Eight patients with negative results on cell block had positive results on pleural biopsy (lung adenocarcinoma in 4, sarcomatoid MPM in 3, and metastatic breast cancer in 1). Two patients with negative results on both cell block and pleural biopsy were diagnosed was sarcomatoid MPM by computed tomography-guided needle biopsy and epithelioid MPM by autopsy. Conclusion Pleural biopsy using flex-rigid pleuroscopy was efficient in the diagnosis of malignant pleural diseases. Flex-rigid pleuroscopy with pleural biopsy and pleural effusion cell block analysis should be considered as the initial diagnostic

  3. 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.

  4. 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.

  5. 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.

  6. Utility of the immunohistochemical detection of FLI-1 expression in round cell and vascular neoplasm using a monoclonal antibody.

    PubMed

    Rossi, Sabrina; Orvieto, Enrico; Furlanetto, Alberto; Laurino, Licia; Ninfo, Vito; Dei Tos, Angelo P

    2004-05-01

    FLI-1 nuclear transcription factor has been proposed as a useful tool in the differential diagnosis of small round cell sarcomas. Recently, FLI-1 has been reported as the first nuclear marker of endothelial differentiation. However, its clinical use has been hampered by major interpretation problems, due to the presence of background staining as well as staining variation between different lots of the same antiserum. In this study, a novel monoclonal antibody raised against the carboxyl terminal of the FLI-1 protein (clone GI146-222, BD Pharmingen) was tested in a series of small round cell and vascular neoplasms. Furthermore, in order to assess FLI-1 specificity, we analyzed its expression in a series of common epithelial and nonepithelial malignancies. In total, 15 Ewing's sarcomas, 10 rhabdomyosarcomas, 5 desmoplastic small round cell tumors, 10 synovial sarcomas, 10 high-grade pleomorphic sarcomas, 10 malignant melanomas, 5 Merkel's carcinomas, 10 colonic adenocarcinomas, 10 breast carcinomas, 10 lung adenocarcinomas, 20 angiosarcomas, 5 epithelioid hemangioendotheliomas, 10 Kaposi's sarcomas and 10 benign hemangiomas, were stained. A strong FLI-1 immunoreactivity was detected in all Ewing's sarcomas and vascular neoplasms, highlighting the high sensitivity of FLI-1 monoclonal antibody. However, 2/5 Merkel's carcinomas and 1/10 malignant melanomas showed a strong nuclear immunostaining, suggesting that FLI-1 may not be so helpful in the differential diagnosis of cutaneous Ewing's sarcoma. In addition, a weak immunoreactivity was found in 3/5 Merkel cell carcinomas, 3/10 synovial sarcomas, 5/10 malignant melanomas, 6/10 lung adenocarcinomas and in 1/10 breast carcinomas. In contrast, all the rhabdomyosarcomas, desmoplastic small round cell tumors, high-grade pleomorphic sarcomas and colonic adenocarcinomas tested were negative. Importantly, in contrast with previous studies, no background staining was observed. Our results indicate that FLI-1 monoclonal antibody

  7. Clinical utility of chromogranin A and octreotide in large cell neuro endocrine carcinoma of the uterine corpus.

    PubMed

    Shahabi, Shohreh; Pellicciotta, Ilenia; Hou, June; Graceffa, Sarah; Huang, Gloria S; Samuelson, Robert N; Goldberg, Gary L

    2011-10-21

    Primary neuroendocrine tumors of the female genital tract have been described in the cervix, ovaries and uterus. Large cell neuroendocrine carcinoma (LCNC) of the uterine corpus is the least common and appears to behave the most aggressively. We report a rare case of a large cell neuroendocrine tumor of the endometrium. These tumors are not well characterized, unlike neuroendocrine tumors of the uterine cervix. Consequently, the optimal management remains still unclear. The treatment of our case consisted of surgery, radiotherapy, chemotherapy, and octreotide. Despite the aggressive treatment, the patient died of disease progression 12 months after the initial diagnosis. We discuss the diagnosis, prognosis, and treatment options for LCNC of the genital tract, and potential future therapeutics.

  8. Clinical utility of chromogranin A and octreotide in large cell neuro endocrine carcinoma of the uterine corpus

    PubMed Central

    Shahabi, Shohreh; Pellicciotta, Ilenia; Hou, June; Graceffa, Sarah; Huang, Gloria S.; Samuelson, Robert N.; Goldberg, Gary L.

    2011-01-01

    Primary neuroendocrine tumors of the female genital tract have been described in the cervix, ovaries and uterus. Large cell neuroendocrine carcinoma (LCNC) of the uterine corpus is the least common and appears to behave the most aggressively. We report a rare case of a large cell neuroendocrine tumor of the endometrium. These tumors are not well characterized, unlike neuroendocrine tumors of the uterine cervix. Consequently, the optimal management remains still unclear. The treatment of our case consisted of surgery, radiotherapy, chemotherapy, and octreotide. Despite the aggressive treatment, the patient died of disease progression 12 months after the initial diagnosis. We discuss the diagnosis, prognosis, and treatment options for LCNC of the genital tract, and potential future therapeutics. PMID:22355496

  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. Transglutaminase 2 and Factor XIII catalyze distinct substrates in differentiating osteoblastic cell line: utility of highly reactive substrate peptides.

    PubMed

    Watanabe, Kazuya; Tsunoda, Kanako; Itoh, Miho; Fukui, Mina; Mori, Hitoshi; Hitomi, Kiyotaka

    2013-01-01

    Differentiated osteoblastic cell line, MC3T3-E1 expresses transglutaminase 2 (TG2) and Factor XIII (FXIII). In previous studies, we identified isozyme-specific and highly reactive glutamine-donor substrate peptides (pepF11KA and pepT26) for each isozyme. Using these peptides, we compared the reaction products with lysine-donor substrates for each isozyme in differentiating MC3T3-E1 cells. By this analysis, distinct substrates for the activated TG2 and FXIII were detected in cultured cellular extract. Possible substrates that incorporated biotin-labeled peptides were further purified using streptavidin-affinity chromatography. Several isozyme-specific substrates were identified by mass spectrometry analysis of the purified fractions. These analyses also indicate the benefit of the substrate peptides for obtaining distinct substrates in a reaction mixture where two isozymes co-exist.

  11. Assessment of electrophile damage in a human brain endothelial cell line utilizing a clickable alkyne analog of 2-chlorohexadecanal.

    PubMed

    Nusshold, Christoph; Üllen, Andreas; Kogelnik, Nora; Bernhart, Eva; Reicher, Helga; Plastira, Ioanna; Glasnov, Toma; Zangger, Klaus; Rechberger, Gerald; Kollroser, Manfred; Fauler, Günter; Wolinski, Heimo; Weksler, Babette B; Romero, Ignacio A; Kohlwein, Sepp D; Couraud, Pierre-Olivier; Malle, Ernst; Sattler, Wolfgang

    2016-01-01

    Peripheral leukocytes aggravate brain damage by releasing cytotoxic mediators that compromise blood-brain barrier function. One of the oxidants released by activated leukocytes is hypochlorous acid (HOCl) that is formed via the myeloperoxidase-H2O2-chloride system. The reaction of HOCl with the endogenous plasmalogen pool of brain endothelial cells results in the generation of 2-chlorohexadecanal (2-ClHDA), a toxic, lipid-derived electrophile that induces blood-brain barrier dysfunction in vivo. Here, we synthesized an alkynyl-analog of 2-ClHDA, 2-chlorohexadec-15-yn-1-al (2-ClHDyA) to identify potential protein targets in the human brain endothelial cell line hCMEC/D3. Similar to 2-ClHDA, 2-ClHDyA administration reduced cell viability/metabolic activity, induced processing of pro-caspase-3 and PARP, and led to endothelial barrier dysfunction at low micromolar concentrations. Protein-2-ClHDyA adducts were fluorescently labeled with tetramethylrhodamine azide (N3-TAMRA) by 1,3-dipolar cycloaddition in situ, which unveiled a preferential accumulation of 2-ClHDyA adducts in mitochondria, the Golgi, endoplasmic reticulum, and endosomes. Thirty-three proteins that are subject to 2-ClHDyA-modification in hCMEC/D3 cells were identified by mass spectrometry. Identified proteins include cytoskeletal components that are central to tight junction patterning, metabolic enzymes, induction of the oxidative stress response, and electrophile damage to the caveolar/endosomal Rab machinery. A subset of the targets was validated by a combination of N3-TAMRA click chemistry and specific antibodies by fluorescence microscopy. This novel alkyne analog is a valuable chemical tool to identify cellular organelles and protein targets of 2-ClHDA-mediated damage in settings where myeloperoxidase-derived oxidants may play a disease-propagating role.

  12. 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.

  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. High sensitivity detection of HPV-16 in SiHa and CaSki cells utilizing FISH enhanced by TSA.

    PubMed

    Adler, K; Erickson, T; Bobrow, M

    1997-01-01

    Detection of integrated human papillomavirus type 16 (HPV-16) DNA in SiHa and CaSki cells was used as a model system to demonstrate sensitivity and resolution of a well defined target. Using 293- to 1987-base polymerase chain reaction (PCR)-synthesized probes to the E6 and E7 open reading frames of HPV-16, several fluorescent in situ hybridization (FISH) detection methods, enhanced with tyramide signal amplification (TSA), were compared. The synthetic probes were biotin labeled by a nick translation method and the hybridized probes were detected by various fluorescent TSA methods using cyanine 3 tyramide, biotinyl tyramide and a biotin TSA Plus reagent. High sensitivity detection in SiHa cells was demonstrated using a 619-base probe to detect two single copies of integrated HPV-16 DNA. In CaSki cells, which contain up to 600 copies of HPV-16 DNA, a 293-base probe was used for detection. The results of these comparisons show that with refinement of TSA methods and reagents, increasing levels of high sensitivity detection can be achieved and that these methods allow subnuclear localization as well.

  15. Human Cells Cultured under Physiological Oxygen Utilize Two Cap-binding Proteins to recruit Distinct mRNAs for Translation.

    PubMed

    Timpano, Sara; Uniacke, James

    2016-05-13

    Translation initiation is a focal point of translational control and requires the binding of eIF4E to the 5' cap of mRNA. Under conditions of extreme oxygen depletion (hypoxia), human cells repress eIF4E and switch to an alternative cap-dependent translation mediated by a homolog of eIF4E, eIF4E2. This homolog forms a complex with the oxygen-regulated hypoxia-inducible factor 2α and can escape translation repression. This complex mediates cap-dependent translation under cell culture conditions of 1% oxygen (to mimic tumor microenvironments), whereas eIF4E mediates cap-dependent translation at 21% oxygen (ambient air). However, emerging evidence suggests that culturing cells in ambient air, or "normoxia," is far from physiological or "normal." In fact, oxygen in human tissues ranges from 1-11% or "physioxia." Here we show that two distinct modes of cap-dependent translation initiation are active during physioxia and act on separate pools of mRNAs. The oxygen-dependent activities of eIF4E and eIF4E2 are elucidated by observing their polysome association and the status of mammalian target of rapamycin complex 1 (eIF4E-dependent) or hypoxia-inducible factor 2α expression (eIF4E2-dependent). We have identified oxygen conditions where eIF4E is the dominant cap-binding protein (21% normoxia or standard cell culture conditions), where eIF4E2 is the dominant cap-binding protein (1% hypoxia or ischemic diseases and cancerous tumors), and where both cap-binding proteins act simultaneously to initiate the translation of distinct mRNAs (1-11% physioxia or during development and stem cell differentiation). These data suggest that the physioxic proteome is generated by initiating translation of mRNAs via two distinct but complementary cap-binding proteins.

  16. Exploring the linkage between cell culture process parameters and downstream processing utilizing a plackett-burman design for a model monoclonal antibody.

    PubMed

    Agarabi, Cyrus D; Chavez, Brittany K; Lute, Scott C; Read, Erik K; Rogstad, Sarah; Awotwe-Otoo, David; Brown, Matthew R; Boyne, Michael T; Brorson, Kurt A

    2017-01-01

    Linkage of upstream cell culture with downstream processing and purification is an aspect of Quality by Design crucial for efficient and consistent production of high quality biopharmaceutical proteins. In a previous Plackett-Burman screening study of parallel bioreactor cultures we evaluated main effects of 11 process variables, such as agitation, sparge rate, feeding regimens, dissolved oxygen set point, inoculation density, supplement addition, temperature, and pH shifts. In this follow-up study, we observed linkages between cell culture process parameters and downstream capture chromatography performance and subsequent antibody attributes. In depth analysis of the capture chromatography purification of harvested cell culture fluid yielded significant effects of upstream process parameters on host cell protein abundance and behavior. A variety of methods were used to characterize the antibody both after purification and buffer formulation. This analysis provided insight in to the significant impacts of upstream process parameters on aggregate formation, impurities, and protein structure. This report highlights the utility of linkage studies in identifying how changes in upstream parameters can impact downstream critical quality attributes. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:163-170, 2017.

  17. Utility of a column-free cell sorting system for separation of plasma cells in multiple myeloma FISH testing in clinical laboratories.

    PubMed

    Shetty, Shashirekha; Siady, Marion; Mallempati, Kalyan C; Wilson, Andrew; Poarch, Jeff; Chandler, Brandon; Gray, Judy; Salama, Mohamed E

    2012-03-01

    Targeted FISH analysis is an essential component of the management of plasma cell myeloma for identification of cytogenetic abnormalities. The purpose of this study was to evaluate the column-free method, RoboSep® (RS), for sorting CD138-expressing cells in bone marrow aspirates. Comparative analysis of column-based and RS methodologies was carried out on 54 paired bone marrow aspirate validation samples from patients undergoing work-up for plasma cell dyscrasia. Abnormalities detected by FISH analysis using an IGH@/CCND1 probe set were seen in 54% with RS, and 44% with column-based. We found a statistically significant difference between the yield of abnormalities detected in paired positive cases (p = 0.0001). An additional 183 consecutive post-validation samples sorted by RS showed recurrent genetic abnormalities in 85/120 (71%) of successfully sorted samples with ≥ 1% plasma cells but in none of 63 samples in which FISH analysis was completed on samples that could not be sorted due to insufficient plasma cells upon cell sorting. The column-free method successfully sorted PC, when present in ≥ 1% of cells, for detection of abnormalities by FISH. Furthermore, our data suggest that FISH analysis should not be performed on samples with an inadequate yield at the cell selection step.

  18. Hepatocyte nuclear factor-1β expression in clear cell adenocarcinomas of the bladder and urethra: diagnostic utility and implications for histogenesis.

    PubMed

    Brimo, Fadi; Herawi, Mehsati; Sharma, Rajni; Netto, Georges J; Epstein, Jonathan I; Illei, Peter B

    2011-11-01

    The histogenesis of clear cell adenocarcinoma of the bladder/urethra is uncertain. Hepatocyte nuclear factor-1β is a homeodomain protein that has been reported to be frequently overexpressed in ovarian clear cell adenocarcinoma in comparison with rare or no expression in other types of epithelial ovarian tumors. We assessed the expression of hepatocyte nuclear factor-1β in a series of 18 clear cell adenocarcinomas of the bladder and urethra and compared it with that of invasive high-grade transitional/urothelial carcinoma (n = 35); adenocarcinomas of the bladder, urethra, and paraurethral glands (n = 21); as well as nephrogenic adenomas of the bladder (n = 8). Staining intensity and extent were evaluated using a 4-tiered grading system (0-3). A case was considered positive for hepatocyte nuclear factor-1β if 10% or more of tumor cells showed at least weak nuclear staining or if any moderate or strong nuclear staining was observed. All 18 clear cell adenocarcinomas exhibited nuclear staining in at least 50% of tumor cells (16 strong, 1 moderate, and 1 weak with focal strong nuclear staining) in comparison with positive nuclear staining (moderate) in 1 of 21 bladder adenocarcinoma, 1 of 35 invasive high-grade transitional/urothelial carcinoma (weak to moderate staining), and 2 of 8 nephrogenic adenomas (1 weak and 1 moderate to strong staining). We concluded that hepatocyte nuclear factor-1β is a useful marker in differentiating clear cell adenocarcinomas of the bladder/urethra from invasive high-grade transitional/urothelial carcinoma and other types of bladder adenocarcinomas and to a lesser extent from nephrogenic adenomas. Hepatocyte nuclear factor-1β is of no diagnostic utility in discriminating primary bladder/urethral clear cell adenocarcinomas from metastatic clear cell adenocarcinomas of the female genital tract to the bladder/urethra. From a histogenesis standpoint, although the expression of hepatocyte nuclear factor-1β in both gynecologic and

  19. Novel migrating mouse neural crest cell assay system utilizing P0-Cre/EGFP fluorescent time-lapse imaging

    PubMed Central

    2011-01-01

    Background Neural crest cells (NCCs) are embryonic, multipotent stem cells. Their long-range and precision-guided migration is one of their most striking characteristics. We previously reported that P0-Cre/CAG-CAT-lacZ double-transgenic mice showed significant lacZ expression in tissues derived from NCCs. Results In this study, by embedding a P0-Cre/CAG-CAT-EGFP embryo at E9.5 in collagen gel inside a culture glass slide, we were able to keep the embryo developing ex vivo for more than 24 hours; this development was with enough NCC fluorescent signal intensity to enable single-cell resolution analysis, with the accompanying NCC migration potential intact and with the appropriate NCC response to the extracellular signal maintained. By implantation of beads with absorbed platelet-derived growth factor-AA (PDGF-AA), we demonstrated that PDGF-AA acts as an NCC-attractant in embryos. We also performed assays with NCCs isolated from P0-Cre/CAG-CAT-EGFP embryos on culture plates. The neuromediator 5-hydroxytryptamine (5-HT) has been known to regulate NCC migration. We newly demonstrated that dopamine, in addition to 5-HT, stimulated NCC migration in vitro. Two NCC populations, with different axial levels of origins, showed unique distribution patterns regarding migration velocity and different dose-response patterns to both 5-HT and dopamine. Conclusions Although avian species predominated over the other species in the NCC study, our novel system should enable us to use mice to assay many different aspects of NCCs in embryos or on culture plates, such as migration, division, differentiation, and apoptosis. PMID:22070366

  20. Utility of α-methylacyl-coenzyme-A racemase (p504s) immunohistochemistry in distinguishing endometrial clear cell carcinomas from serous and endometrioid carcinomas.

    PubMed

    Fadare, Oluwole; Parkash, Vinita; Gwin, Katja; Hanley, Krisztina Z; Jarboe, Elke A; Liang, Sharon X; Quick, Charles M; Zheng, Wenxin; Rawish, Kojo R; Hecht, Jonathan L; Desouki, Mohamed M

    2013-12-01

    The expression of α-methylacyl-coenzyme-A racemase (AMACR) has previously been reported in 75% to 100% of urethral/bladder clear cell carcinomas, tumors that are known to display broad phenotypic overlap with their identically named müllerian counterparts. Herein, we assess the utility of AMACR in distinguishing endometrial clear cell carcinomas (CCCs) from endometrial serous carcinomas (ESCs) and endometrial endometrioid carcinomas (EECs). A total of 111 endometrial carcinomas in a tissue microarray, including 49 CCCs, 13 ESCs, and 49 EECs, were assessed for AMACR immunoreactivity, with results scored semiquantitatively (scores 0, 1+, 2+, 3+ for 0%, 1%-5%, 6%-50%, >50% immunoreactive cells, respectively). Fifty (45%) of the 111 carcinomas were AMACR positive, with the following score distribution: CCC: 0 (n = 12), 1+ (n = 12), 2+ (n = 3), 3+ (n = 22); EEC: 0 (n = 38), 1+ (n = 4), 2+ (n = 4), 3+ (n = 3); ESC: 0 (n = 11), 1+ (n = 1), 2+ (n = 0), 3+ (n = 1). AMACR expression was significantly more frequent in CCC (75%) than in ESC (15%) or EEC (22%); P < .0001. The sensitivity and specificity of AMACR expression in classifying a carcinoma as CCC were 0.75 (95% confidence interval [CI], 0.61-0.86) and 0.79 (95% CI, 0.66-0.88), respectively, with an odds ratio of 11.62 (95% CI, 5-28; P < .001) and an area under the curve of 0.79 (95% CI, 0.68-0.88). These findings indicate that AMACR expression is strongly associated with CCC and displays a relatively robust diagnostic test performance. However, its practical utility may be limited by the focal nature of its expression in 32% of the AMACR-positive CCC cases as well as its expression in 15% to 22% of the non-CCC histotypes.

  1. 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

  2. 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

  3. Modular Utilization of Distal cis-Regulatory Elements Controls Ifng Gene Expression in T Cells Activated by Distinct Stimuli

    PubMed Central

    Balasubramani, Anand; Shibata, Yoichiro; Crawford, Gregory E.; Baldwin, Albert S.; Hatton, Robin D.; Weaver, Casey T.

    2010-01-01

    SUMMARY Distal cis-regulatory elements play essential roles in the T lineage-specific expression of cytokine genes. We have mapped interactions of three transacting factors – NF-κB, STAT4 and T-bet – with cis elements in the Ifng locus. We find that RelA is critical for optimal Ifng expression and is differentially recruited to multiple elements contingent upon T cell receptor (TCR) or interleukin-12 (IL-12) plus IL-18 signaling. RelA recruitment to at least four elements is dependent on T-bet-dependent remodeling of the Ifng locus and co-recruitment of STAT4. STAT4 and NF-κB therefore cooperate at multiple cis elements to enable NF-κB–dependent enhancement of Ifng expression. RelA recruitment to distal elements was similar in Th1 and Tc1 effector cells, although T-bet was dispensable in CD8 effectors. These results support a model of Ifng regulation in which distal cis-regulatory elements differentially recruit key transcription factors in a modular fashion to initiate gene transcription induced by distinct activation signals. PMID:20643337

  4. 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.

  5. 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

    2016-08-16

    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.

  6. 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

  7. Promotion of glucose utilization by insulin enhances granulosa cell proliferation and developmental competence of porcine oocyte grown in vitro.

    PubMed

    Itami, Nobuhiko; Munakata, Yasuhisa; Shirasuna, Koumei; Kuwayama, Takehito; Iwata, Hisataka

    2017-02-01

    In vitro culture of the oocyte granulosa cell complexes (OGCs) from early antral follicles (EAFs) shows granulosa cell (GC) proliferation, but to a lesser extent than that observed in vivo during follicle development. As the number of GCs closely relates to energy sufficiency of the oocytes, enhancement of GC proliferation influences oocyte development. GC proliferation depends on glycolysis and insulin-mediated AKT/mTOR signaling pathway; therefore, addition of culture medium containing insulin and glucose may potentially promote GC proliferation and hence improve oocyte development. In the present study, we assessed the effect of exogenous insulin and glucose concentration on GC proliferation and oocyte energy status as well as developmental abilities of porcine oocytes grown in vitro. In the presence of 5.5 mM of glucose (Low), a comparison of 10 versus 20 μg/ml insulin showed that high insulin enhanced GC proliferation but exhausted glucose from the medium, which resulted in low energy status including lipid and adenosine triphosphate of the oocyte. Whereas, in the presence of 20 μg/ml insulin, medium with 11 mM glucose (High) enhanced GC proliferation and oocyte energy status as well as developmental ability up to the blastocyst stage. Considering that there was no difference in OGCs development observed with medium (10 μg/ml insulin) containing 5.5 versus 11 mM glucose, we concluded that the combination of high insulin and glucose enhanced GC proliferation and energy status of oocytes as well as the developmental ability of the oocytes grown in vitro.

  8. Stimulation of apoptotic pathways in liver cancer cells: An alternative perspective on the biocompatibility and the utility of biomedical glasses.

    PubMed

    Kilcup, Nancy; Gaynard, Seán; Werner-Zwanziger, Ulrike; Tonkopi, Elena; Hayes, Jessica; Boyd, Daniel

    2016-05-01

    A host of research opportunities with innumerable clinical applications are open to biomedical glasses if one considers their potential as therapeutic inorganic ion delivery systems. Generally, applications have been limited to repair and regeneration of hard tissues while compositions are largely constrained to the original bioactive glass developed in the 1960s. However, in oncology applications the therapeutic paradigm shifts from repair to targeted destruction. With this in mind, the composition-structure-property-function relationships of vanadium-containing zinc-silicate glasses (0.51SiO2-0.29Na2O-(0.20-X)ZnO-XV2O5, 0 ≤ X ≤ 0.09) were characterized in order to determine their potential as therapeutic inorganic ion delivery systems. Increased V2O5mole fraction resulted in a linear decrease in density and glass transition temperature (Tg).(29)Si MAS NMR peak maxima shifted upfield while(51)V MAS NMR peak maxima were independent of V2O5content and overlapped well with the spectra NaVO3 Increased V2O5mole fraction caused ion release to increase. When human liver cancer cells, HepG2, were exposed to these ions they demonstrated a concentration-dependent cytotoxic response, mediated by apoptosis. This work demonstrates that the zinc-silicate system studied herein is capable of delivering therapeutic inorganic ions at concentrations that induce apoptotic cell death and provide a simple means to control therapeutic inorganic ion delivery.

  9. 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.

  10. The utility of p16 immunostaining in fine needle aspiration in p16-positive head and neck squamous cell carcinoma.

    PubMed

    Xu, Bin; Ghossein, Ronald; Lane, Jason; Lin, Oscar; Katabi, Nora

    2016-08-01

    Many patients with human papillomavirus (HPV)-related head and neck squamous cell carcinoma present initially with cervical nodal metastasis. Fine needle aspiration (FNA) of the nodal disease might be the only diagnostic material available for p16 immunohistochemistry (IHC) and HPV testing. The current study aims to evaluate p16 IHC in FNA and establish guidelines for its interpretation. The percentage and intensity of p16 IHC staining were examined in 60 matched FNA and surgical cases. Cytomorphologic features were included in the analysis. p16 IHC staining was correlated with the results seen in the surgical specimens and with HPV in situ hybridization (ISH). Analysis of different thresholds demonstrated that the threshold of 10% p16 tumor cell positivity had the best overall concordance rate with surgical p16 IHC (κ = 0.650) and with FNA HPV-ISH (κ = 0.714). Applying the recommended p16 positivity threshold for surgical specimens (70%) on FNA materials resulted in low sensitivity (39%) and low negative predictive value (26%). In comparison with p16 IHC in surgical specimens, 6/46 FNA cases (13%) were falsely negative for p16. All 6 cases were associated with necrotic background, two (33%) lacked large tumor clusters, and one (17%) had low cellularity. The recommended threshold for p16 IHC on surgical specimens should not be used in cytology materials. The cutoff value for p16 immunostain in FNA specimens showing best results in our series is 10%. When p16 IHC is negative in FNA specimens, a repeat stain on a surgical specimen is recommended to avoid a false-negative diagnosis.

  11. 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

  12. 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

  13. The addition of decision support into computerized physician order entry reduces red blood cell transfusion resource utilization in the intensive care unit.

    PubMed

    Fernández Pérez, Evans R; Winters, Jeffrey L; Gajic, Ognjen

    2007-07-01

    Computerized physician order entry (CPOE) has the potential for cost containment in critically ill patients through practice standardization and elimination of unnecessary interventions. Previous study demonstrated the beneficial short-term effect of adding a decision support for red blood cell (RBC) transfusion into the hospital CPOE. We evaluated the effect of such intervention on RBC resource utilization during the two-year study period. From the institutional APACHE III database we identified 2,200 patients with anemia, but no active bleeding on admission: 1,100 during a year before and 1,100 during a year after the intervention. The mean number of RBC transfusions per patient decreased from 1.5 +/- 1.9 units to 1.3 +/- 1.8 units after the intervention (P = 0.045). RBC transfusion cost decreased from $616,442 to $556,226 after the intervention. Hospital length of stay and adjusted hospital mortality did not differ before and after protocol implementation. In conclusion, the implementation of an evidenced-based decision support system through a CPOE can decrease RBC transfusion resource utilization in critically ill patients.

  14. Safety and Effectiveness of Bone Marrow Cell Concentrate in the Treatment of Chronic Critical Limb Ischemia Utilizing a Rapid Point-of-Care System

    PubMed Central

    Gupta, Saniya; Sethi, Dalip; Powell, Richard J.; Harris, Kenneth Lee; Jungla, Nungshi; Arambam, Priyadarshini; Kaul, Upendra; Seth, Ashok; Bukhari, Suhail

    2017-01-01

    Critical limb ischemia (CLI) is the end stage of lower extremity peripheral vascular disease (PVD) in which severe obstruction of blood flow results in ischemic rest pain, ulcers and/or gangrene, and a significant risk of limb loss. This open-label, single-arm feasibility study evaluated the safety and therapeutic effectiveness of autologous bone marrow cell (aBMC) concentrate in revascularization of CLI patients utilizing a rapid point-of-care device. Seventeen (17) no-option CLI patients with ischemic rest pain were enrolled in the study. Single dose of aBMC, prepared utilizing an intraoperative point-of-care device, the Res-Q™ 60 BMC system, was injected intramuscularly into the afflicted limb and patients were followed up at regular intervals for 12 months. A statistically significant improvement in Ankle Brachial Index (ABI), Transcutaneous Oxygen Pressure (TcPO2), mean rest pain and intermittent claudication pain scores, wound/ ulcer healing, and 6-minute walking distance was observed following aBMC treatment. Major amputation-free survival (mAFS) rate and amputation-free rates (AFR) at 12 months were 70.6% and 82.3%, respectively. In conclusion, aBMC injections were well tolerated with improved tissue perfusion, confirming the safety, feasibility, and preliminary effectiveness of aBMC treatment in CLI patients. PMID:28194186

  15. Safety and Effectiveness of Bone Marrow Cell Concentrate in the Treatment of Chronic Critical Limb Ischemia Utilizing a Rapid Point-of-Care System.

    PubMed

    Ponemone, Venkatesh; Gupta, Saniya; Sethi, Dalip; Suthar, Manish; Sharma, Monika; Powell, Richard J; Harris, Kenneth Lee; Jungla, Nungshi; Arambam, Priyadarshini; Kaul, Upendra; Seth, Ashok; Bukhari, Suhail

    2017-01-01

    Critical limb ischemia (CLI) is the end stage of lower extremity peripheral vascular disease (PVD) in which severe obstruction of blood flow results in ischemic rest pain, ulcers and/or gangrene, and a significant risk of limb loss. This open-label, single-arm feasibility study evaluated the safety and therapeutic effectiveness of autologous bone marrow cell (aBMC) concentrate in revascularization of CLI patients utilizing a rapid point-of-care device. Seventeen (17) no-option CLI patients with ischemic rest pain were enrolled in the study. Single dose of aBMC, prepared utilizing an intraoperative point-of-care device, the Res-Q™ 60 BMC system, was injected intramuscularly into the afflicted limb and patients were followed up at regular intervals for 12 months. A statistically significant improvement in Ankle Brachial Index (ABI), Transcutaneous Oxygen Pressure (TcPO2), mean rest pain and intermittent claudication pain scores, wound/ ulcer healing, and 6-minute walking distance was observed following aBMC treatment. Major amputation-free survival (mAFS) rate and amputation-free rates (AFR) at 12 months were 70.6% and 82.3%, respectively. In conclusion, aBMC injections were well tolerated with improved tissue perfusion, confirming the safety, feasibility, and preliminary effectiveness of aBMC treatment in CLI patients.

  16. Utilization of signal transduction pathway by the human T-cell leukemia virus type I transcriptional activator tax

    SciTech Connect

    Tan, Tsehua; Jia, R.; Goeder, R.G. )

    1989-09-01

    The human T-cell leukemia virus type I (HTLV-I) trans-activator (tax)-inducible enhancer was localized to three copies of 21-base-pair repeats within the long terminal repeat. Interestingly, the TGACG motif found in the center of the 21-base-pair tax-responsive element (TRE) is also present in the cyclic AMP (cAMP)-responsive elements (CREs) and activating transcription factor (ATF)-binding sites. In this study, the authors demonstrate that the three TRE-binding proteins, TREB-1, TREB-2, and TREB-3, also bind to various CREs and ATF-binding sites and that the TREs can confer upon a heterologous promoter responsiveness to various inducing agents, including tax, cAMP, and E1a. Furthermore, the transcriptional activation of the HTLV-I promoter by tax can be inhibited by several protein kinase inhibitors, including sangivamycin. The results indicate that the TREs, CREs, and ATF-binding sites are similar cis-acting elements and further suggest (i) that the transcriptional activation of the HTLV-I promoter by tax involves the action of a protein kinase an (ii) that induction by tax, cAMP, and E1a might be mediated by distinct factors or kinases.

  17. An ultrasensitive squamous cell carcinoma antigen biosensing platform utilizing double-antibody single-channel amplification strategy.

    PubMed

    Ren, Xiang; Wu, Dan; Wang, Yuhuan; Zhang, Yunhui; Fan, Dawei; Pang, Xuehui; Li, Yueyun; Du, Bin; Wei, Qin

    2015-10-15

    A novel electrochemical immunosensor was developed for ultrasensitive detection of squamous cell carcinoma antigen (SCCA), which was based on the double-antibody single-channel amplification strategy. For the first time, human immunoglobulin antibody (anti-HIgG) was used as the supporting framework to amplify the loading quantity of SCCA antibody (anti-SCCA). In this strategy, SCCA can be detected without using mesoporous nanometers to amplify the signal. In addition, Pd icosahedrons were first used as the connecter to immobilize the antibodies and strengthen the sensitivity. Only one touch point exists under the limited condition between a sphere and another shape in geometry, thus the Pd icosahedron is an excellent candidate as the role of connecter. Gold nanoparticles (Au NPs) decorated with mercapto-functionalized graphene sheets (Au@GS) were synthesized as the transducing materials. The fabricated immunosensor exhibited an excellent detection limit of 2.8 pg/mL and wide linear range of 0.01-5 ng/mL. This kind of immunosensor would provide a potential application in clinical diagnosis.

  18. 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.

  19. 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.

  20. Utilization of Collaborative Practice Agreements between Physicians and Pharmacists as a Mechanism to Increase Capacity to Care for Hematopoietic Stem Cell Transplant Recipients

    PubMed Central

    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

    2013-01-01

    Survival following 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

  1. 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.

  2. Clinical utility of erlotinib for the treatment of non-small-cell lung cancer in Japanese patients: current evidence

    PubMed Central

    Togashi, Yosuke; Hayashi, Hidetoshi; Nakagawa, Kazuhiko; Nishio, Kazuto

    2014-01-01

    Gefitinib, an epidermal growth factor tyrosine kinase inhibitor (EGFR-TKI), has been approved in Japan for the treatment of patients with advanced non-small-cell lung cancer (NSCLC) based on Phase II clinical trials since 2002. Erlotinib, another EGFR-TKI, was also approved a few years thereafter. In 2004, activating mutations in the EGFR gene were discovered to be a predictive biomarker for EGFR-TKI treatment, and gefitinib, which is not effective for patients with EGFR wild-type NSCLC, has since been used only in patients with EGFR-mutated NSCLC. In contrast, erlotinib is potentially effective for the treatment of EGFR wild-type NSCLC. Similar to gefitinib, erlotinib is also effective for EGFR-mutated NSCLC and has been used as an initial treatment for patients with advanced EGFR-mutated NSCLC. Both gefitinib and erlotinib can be used in a Japanese clinical setting. The approved daily dose of erlotinib (150 mg) is equal to the maximum tolerated dose of erlotinib. In contrast, the daily dose of gefitinib has been set at 250 mg, which is approximately one-third of the maximum tolerated dose of gefitinib. Accordingly, a higher serum concentration can be achieved using erlotinib, compared with gefitinib. This advantage can be applied to the treatment of central nervous system metastases (brain metastasis and carcinomatous meningitis), the treatment of which is complicated by the difficulty drugs have penetrating the blood–brain barrier. Although patients with EGFR-mutated NSCLC respond dramatically to EGFR-TKIs, some patients have a poor response and the majority eventually undergo disease progression. To overcome such resistance, several novel treatment strategies, such as combination therapy and next-generation EGFR-TKIs, have been attempted. PMID:25114510

  3. Mesenchymal stem cell seeding promotes reendothelialization of the endovascular stent.

    PubMed

    Wu, Xue; Wang, Guixue; Tang, Chaojun; Zhang, Dechuan; Li, Zhenggong; Du, Dingyuan; Zhang, Zhengcai

    2011-09-01

    This study is designed to make a novel cell seeding stent and to evaluate reendothelialization and anti-restenosis after the stent implantation. In comparison with cell seeding stents utilized in previous studies, Mesenchymal stem cells (MSCs) have advantages on promoting of issue repair. Thus it was employed to improve the reendothelialization effects of endovascular stent in present work. MSCs were isolated by density gradient centrifugation and determined as CD29(+) CD44(+) CD34(-) cells by immunofluorescence and immunocytochemistry; gluten and polylysine coated stents were prepared by ultrasonic atomization spray, and MSCs seeded stents were made through rotation culture according to the optimized conditions that were determined in previous studies. The results from animal experiments, in which male New Zealand white rabbits were used, show that the reendothelialization of MSCs coated stents can be completed within one month; in comparison with 316L stainless steel stents (316L SS stents) and gluten and polylysine coated stents, the intimal hyperplasia and in-stent restenosis are significantly inhibited by MSCs coated stents. Endovascular stent seeded with MSCs promotes reendothelialization and inhibits the intimal hyperplasia and in-stent restenosis compared with the 316L SS stents and the gluten and polylysine coated stents.

  4. Degradative capacities and 16S rRNA-targeted whole-cell hybridization of sulfate-reducing bacteria in an anaerobic enrichment culture utilizing alkylbenzenes from crude oil.

    PubMed Central

    Rabus, R; Fukui, M; Wilkes, H; Widdle, F

    1996-01-01

    A mesophilic sulfate-reducing enrichment culture growing anaerobically on crude oil was used as a model system to study which nutritional types of sulfate-reducing bacteria may develop on original petroleum constituents in oil wells, tanks, and pipelines. Chemical analysis of oil hydrocarbons during growth revealed depletion of toluene and o-xylene within 1 month and of m-xylene, o-ethyltoluene, m-ethyltoluene, m-propyltoluene, and m-isopropyltoluene within approximately 2 months. In anaerobic counting series, the highest numbers of CFU (6 x 10(6) to 8 x 10(6) CFU ml-1) were obtained with toluene and benzoate. Almost the same numbers were obtained with lactate, a substrate often used for detection of the vibrio-shaped, incompletely oxidizing Desulfovibrio sp. In the present study, however, lactate yielded mostly colonies of oval to rod-shaped, completely oxidizing, sulfate-reducing bacteria which were able to grow slowly on toluene or crude oil. Desulfovibrio species were detected only at low numbers (3 x 10(5) CFU ml-1). In agreement with this finding, a fluorescently labeled, 16S rRNA-targeted oligonucleotide probe described in the literature as specific for members of the Desulfovibrionaceae (suggested family) hybridized only with a small portion (< 5%) of the cells in the enrichment culture. These results are consistent with the observation that known Desulfovibrio species do not utilize aromatic hydrocarbons, the predominant substrates in the enrichment culture. All known sulfate-reducing bacteria which utilize aromatic compounds belong to a separate branch, the Desulfobacteriaceae (suggested family). Most members of this family are complete oxidizers. For specific hybridization with members of this branch, the probe had to be modified by a nucleotide exchange. Indeed, this modified probe hybridized with more than 95% of the cells in the enrichment culture. The results show that completely oxidizing, alkylbenzene-utilizing sulfate-reducing bacteria rather than

  5. High-performance Förster resonance energy transfer (FRET)-based dye-sensitized solar cells: rational design of quantum dots for wide solar-spectrum utilization.

    PubMed

    Lee, Eunwoo; Kim, Chanhoi; Jang, Jyongsik

    2013-07-29

    High-performance Förster resonance energy transfer (FRET)-based dye-sensitized solar cells (DSSCs) have been successfully fabricated through the optimized design of a CdSe/CdS quantum-dot (QD) donor and a dye acceptor. This simple approach enables quantum dots and dyes to simultaneously utilize the wide solar spectrum, thereby resulting in high conversion efficiency over a wide wavelength range. In addition, major parameters that affect the FRET interaction between donor and acceptor have been investigated including the fluorescent emission spectrum of QD, and the content of deposited QDs into the TiO2 matrix. By judicious control of these parameters, the FRET interaction can be readily optimized for high photovoltaic performance. In addition, the as-synthesized water-soluble quantum dots were highly dispersed in a nanoporous TiO2 matrix, thereby resulting in excellent contact between donors and acceptors. Importantly, high-performance FRET-based DSSCs can be prepared without any infrared (IR) dye synthetic procedures. This novel strategy offers great potential for applications of dye-sensitized solar cells.

  6. 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.

  7. 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.

  8. Phaeobacter sp. strain Y4I utilizes two separate cell-to-cell communication systems to regulate production of the antimicrobial indigoidine.

    PubMed

    Cude, W Nathan; Prevatte, Carson W; Hadden, Mary K; May, Amanda L; Smith, Russell T; Swain, Caleb L; Campagna, Shawn R; Buchan, Alison

    2015-02-01

    The marine roseobacter Phaeobacter sp. strain Y4I synthesizes the blue antimicrobial secondary metabolite indigoidine when grown in a biofilm or on agar plates. Prior studies suggested that indigoidine production may be, in part, regulated by cell-to-cell communication systems. Phaeobacter sp. strain Y4I possesses two luxR and luxI homologous N-acyl-L-homoserine lactone (AHL)-mediated cell-to-cell communication systems, designated pgaRI and phaRI. We show here that Y4I produces two dominantAHLs, the novel monounsaturated N-(3-hydroxydodecenoyl)-L-homoserine lactone (3OHC(12:1)-HSL) and the relatively common N-octanoyl-L-homoserine lactone (C8-HSL), and provide evidence that they are synthesized by PhaI and PgaI, respectively.A Tn5 insertional mutation in either genetic locus results in the abolishment (pgaR::Tn5) or reduction (phaR::Tn5) of pigment production. Motility defects and denser biofilms were also observed in these mutant backgrounds, suggesting an overlap in the functional roles of these systems. Production of the AHLs occurs at distinct points during growth on an agar surface and was determined by isotope dilution high-performance liquid chromatography–tandem mass spectrometry (ID-HPLC-MS/MS) analysis.Within 2 h of surface inoculation, only 3OHC(12:1)-HSL was detected in agar extracts. As surface-attached cells became established (at approximately 10 h), the concentration of 3OHC(12:1)-HSL decreased, and the concentration of C8-HSL increased rapidly over 14 h.After longer (>24-h) establishment periods, the concentrations of the two AHLs increased to and stabilized at approximately 15 nM and approximately 600 nM for 3OHC12:1-HSL and C8-HSL, respectively. In contrast, the total amount of indigoidine increased steadily from undetectable to 642 Mby 48 h. Gene expression profiles of the AHL and indigoidine synthases (pgaI, phaI, and igiD) were consistent with their metabolite profiles. These data provide evidence that pgaRI and phaRI play overlapping roles

  9. 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

  10. miRNA in situ hybridization in circulating tumor cells--MishCTC.

    PubMed

    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

    2015-03-17

    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.

  11. Enrichment of circulating melanoma cells (CMCs) using negative selection from patients with metastatic melanoma

    PubMed Central

    Joshi, Powrnima; Jacobs, Barbara; Derakhshan, Adeeb; Moore, Lee R.; Elson, Paul; Triozzi, Pierre L.; Borden, Ernest; Zborowski, Maciej

    2014-01-01

    Circulating tumor cells have emerged as prognostic biomarkers in the treatment of metastatic cancers of epithelial origins viz., breast, colorectal and prostate. These tumors express Epithelial Cell Adhesion Molecule (EpCAM) on their cell surface which is used as an antigen for immunoaffinity capture. However, EpCAM capture technologies are of limited utility for non-epithelial cancers such as melanoma. We report a method to enrich Circulating Melanoma Cells (CMCs) that does not presuppose malignant cell characteristics. CMCs were enriched by centrifugation of blood samples from healthy (N = 10) and patient (N = 11) donors, followed by RBC lysis and immunomagnetic depletion of CD45-positive leukocytes in a specialized magnetic separator. CMCs were identified by immunocytochemistry using Melan-A or S100B as melanoma markers and enumerated using automated microscopy image analyses. Separation was optimized for maximum sensitivity and recovery of CMCs. Our results indicate large number of CMCs in Stage IV melanoma patients. Analysis of survival suggested a trend toward decreased survival with increased number of CMCs. Moreover, melanoma-associated miRs were found to be higher in CMC-enriched fractions in two patients when compared with the unseparated samples, validating this method as applicable for molecular analyses. Negative selection is a promising approach for isolation of CMCs and other EpCAM -negative CTCs, and is amenable to molecular analysis of CMCs. Further studies are required to validate its efficacy at capturing specific circulating cells for genomic analysis, and xenograft studies. PMID:24811334

  12. Enrichment of circulating melanoma cells (CMCs) using negative selection from patients with metastatic melanoma.

    PubMed

    Joshi, Powrnima; Jacobs, Barbara; Derakhshan, Adeeb; Moore, Lee R; Elson, Paul; Triozzi, Pierre L; Borden, Ernest; Zborowski, Maciej

    2014-05-15

    Circulating tumor cells have emerged as prognostic biomarkers in the treatment of metastatic cancers of epithelial origins viz., breast, colorectal and prostate. These tumors express Epithelial Cell Adhesion Molecule (EpCAM) on their cell surface which is used as an antigen for immunoaffinity capture. However, EpCAM capture technologies are of limited utility for non-epithelial cancers such as melanoma. We report a method to enrich Circulating Melanoma Cells (CMCs) that does not presuppose malignant cell characteristics. CMCs were enriched by centrifugation of blood samples from healthy (N = 10) and patient (N = 11) donors, followed by RBC lysis and immunomagnetic depletion of CD45-positive leukocytes in a specialized magnetic separator. CMCs were identified by immunocytochemistry using Melan-A or S100B as melanoma markers and enumerated using automated microscopy image analyses. Separation was optimized for maximum sensitivity and recovery of CMCs. Our results indicate large number of CMCs in Stage IV melanoma patients. Analysis of survival suggested a trend toward decreased survival with increased number of CMCs. Moreover, melanoma-associated miRs were found to be higher in CMC-enriched fractions in two patients when compared with the unseparated samples, validating this method as applicable for molecular analyses. Negative selection is a promising approach for isolation of CMCs and other EpCAM -negative CTCs, and is amenable to molecular analysis of CMCs. Further studies are required to validate its efficacy at capturing specific circulating cells for genomic analysis, and xenograft studies.

  13. 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.

  14. Neuromuscular junction formation between human stem cell-derived motoneurons and human skeletal muscle in a defined system.

    PubMed

    Guo, Xiufang; Gonzalez, Mercedes; Stancescu, Maria; Vandenburgh, Herman H; Hickman, James J

    2011-12-01

    Functional in vitro models composed of human cells will constitute an important platform in the next generation of system biology and drug discovery. This study reports a novel human-based in vitro Neuromuscular Junction (NMJ) system developed in a defined serum-free medium and on a patternable non-biological surface. The motoneurons and skeletal muscles were derived from fetal spinal stem cells and skeletal muscle stem cells. The motoneurons and skeletal myotubes were completely differentiated in the co-culture based on morphological analysis and electrophysiology. NMJ formation was demonstrated by phase contrast microscopy, immunocytochemistry and the observation of motoneuron-induced muscle contractions utilizing time-lapse recordings and their subsequent quenching by d-Tubocurarine. Generally, functional human based systems would eliminate the issue of species variability during the drug development process and its derivation from stem cells bypasses the restrictions inherent with utilization of primary human tissue. This defined human-based NMJ system is one of the first steps in creating functional in vitro systems and will play an important role in understanding NMJ development, in developing high information content drug screens and as test beds in preclinical studies for spinal or muscular diseases/injuries such as muscular dystrophy, Amyotrophic lateral sclerosis and spinal cord repair.

  15. 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.

  16. SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry.

    PubMed

    Larsen, Jakob V; Petersen, Claus M

    2017-01-06

    The heterodimeric cytokine Cardiotrophin-like Cytokine:Cytokine-like Factor-1 (CLC:CLF-1) targets the glycosylphosphatidylinositol (GPI)-anchored CNTFRα to form a trimeric complex that subsequently recruits glycoprotein 130/Leukemia Inhibitory Factor Receptor-β (gp130/LIFRβ) for signaling. Both CLC and CNTFRα are necessary for signaling but so far CLF-1 has only been known as a putative facilitator of CLC secretion. However, it has recently been shown that CLF-1 contains three binding sites: one for CLC; one for CNTFRα (that may promote assembly of the trimeric complex); and one for the endocytic receptor sorLA. The latter site provides high affinity binding of CLF-1, CLC:CLF-1, as well as the trimeric (CLC:CLF-1:CNTFRα) complex to sorLA, and in sorLA-expressing cells the soluble ligands CLF-1 and CLC:CLF-1 are rapidly taken up and internalized. In cells co-expressing CNTFRα and sorLA, CNTFRα first binds CLC:CLF-1 to form a membrane-associated trimeric complex, but it also connects to sorLA via the free sorLA-binding site in CLF-1. As a result, CNTFRα, which has no capacity for endocytosis on its own, is tugged along and internalized by the sorLA-mediated endocytosis of CLC:CLF-1. The present protocol describes the experimental procedures used to demonstrate i) the sorLA-mediated and CLC:CLF-1-dependent downregulation of surface-membrane CNTFRα expression; ii) sorLA-mediated endocytosis and lysosomal targeting of CNTFRα; and iii) the lowered cellular response to CLC:CLF-1-stimulation upon sorLA-mediated downregulation of CNTFRα.

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

    SciTech Connect

    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 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. Lastly, the identification

  18. 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.

  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. 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.

  1. Combined prognostic utility of white blood cell count, plasma glucose, and glomerular filtration rate in patients undergoing primary stent placement for acute myocardial infarction.

    PubMed

    Kosuge, Masami; Kimura, Kazuo; Morita, Satoshi; Kojima, Sunao; Sakamoto, Tomohiro; Ishihara, Masaharu; Asada, Yujiro; Tei, Chuwa; Miyazaki, Shunichi; Sonoda, Masahiro; Tsuchihashi, Kazufumi; Yamagishi, Masakazu; Shirai, Mutsunori; Hiraoka, Hisatoyo; Honda, Takashi; Ogata, Yasuhiro; Ogawa, Hisao

    2009-02-01

    Although high white blood cell (WBC) count and plasma glucose (PG) and low glomerular filtration rate (GFR) on admission have been associated with poor outcomes after acute myocardial infarction (AMI), the combined prognostic utility of these 3 variables was unclear. The association of WBC count, PG, and GFR on admission to in-hospital outcomes was examined in 2,633 patients who underwent primary stent placement for ST-segment elevation AMI within 48 hours after symptom onset. In-hospital mortality progressively increased as the number of the variables of high WBC count (> or =11,120/microl; upper tertile), high PG (> or =10.4 mmol/L; upper tertile), and low GFR (< or =60 ml/min/1.73 m(2); lower tertile) increased. Patients with all 3 variables had a strikingly higher in-hospital mortality rate (25.9%). After adjusting for baseline characteristics, multivariate analysis showed that compared with patients who had none of these variables, odds ratios for in-hospital mortality were 1.63 (95% confidence interval [CI] 0.88 to 3.03, p = 0.12) in patients with only 1 variable, 2.33 (95% CI 1.28 to 3.96, p = 0.047) in those with 2 variables, and 6.16 (95% CI 2.98 to 12.6, p <0.001) in those with all 3 variables. In conclusion, combined evaluation of WBC count, PG, and GFR on admission was a simple and useful method for the early prediction of risk of in-hospital death in patients undergoing primary stent placement for ST-segment elevation AMI.

  2. [A study on the appropriate fixation for the procedures for the better preservation of cellular antigenicity and morphology of the blood smear in immunocytochemistry: an improvement of the immunostain technique using alkaline-phosphatase (ALP) as a labeling enzyme].

    PubMed

    Aoki, J; Sasaki, N; Hino, N; Nanba, K

    1991-01-01

    The authors previously reported a new coloration method which utilized hexazotized newfuchsin as a coupler for the immuno-enzyme-cytochemistry. This procedure used alkaline-phosphatase (ALP) as the labeling enzyme. The insolubility of the reaction product to organic solvents made it possible to prepare permanent slides. However, this suffered from several drawbacks, due to the fixation procedures, in the preservation of better morphology and antigenicity of the cell. The present study was undertaken to overcome such problems by modifying the fixation procedure. The study utilized twenty monoclonal and polyclonal antibodies commonly used in immunohematological staining. Various fixative solutions and timing of fixation were evaluated. The results indicated that; 1) the best fixative solution was a mixture of buffered paraformaldehyde (PFA) and acetone (10 ml 40% PFA solution, 10 ml pH 6.6 0.02 M phosphate buffer, 20 ml distilled water, 60 ml acetone, with pH adjusted to 6.6-7.4 with HCl) and 2) the fixation should be performed just before the immunostain. The results further showed that unstained smear slides, when freshly air dried and stored in a desicator, could maintain various differentiation antigens (CD2, 3, 4, 5, 8, 10, 14, 15, 16, 19, 25, L26, HLA-DR) for at least 4 weeks without any change in the immuno-reactivity. Thus, we conclude that this improved fixation procedure is an optimum fixative and should be used in routine application of the immunostain method for blood smears.

  3. Neuronal differentiation of PC12 and embryonic stem cells in two- and three-dimensional in vitro culture.

    PubMed

    Sadri, Soheil; Khazaei, Mozafar; Ghanbari, Ali; Khazaei, Mohammad Rasool; Shah, Palak

    2014-04-01

    The quality of neuronal differentiation and reduction in apoptosis that occurred in two-dimensional (2D) and three-dimensional (3D) culture conditions is compared. PC12 and embryonic stem cells are two commonly utilized cell lines for the study of neuronal regeneration. These cells were induced to neuronally differentiate by adding NGF and retinoic acid respectively. Total neurite length and expression of neuronal markers (MAP-2 and beta-tubulin) was assessed by morphometry and immunocytochemistry. Also, TUNEL assay was used to detect apoptosis. Upon exposure to a differentiation media in the 3D fibrin gel, PC12 and embryonic stem cells stopped dividing, had increased adhesion to the substratum, extended neurite processes and expressed neuronal markers. The same results, however, were not observedwith the 2D culture. Also, the apoptosis index performed by TUNEL a ss ay demonstrated a reduction in th e degree of apoptosis in the 3D culture compared to 2D culture. Fibrin matrix supports growth and n euronal differentiation of PC12 andembryonic stem cells. In addition, the 3D culture enhanced cellular resistance to apoptosis when compared to the 2D culture. It appears as if a 3D culture system may offer a better technique for future neuronal tissue engineering investigations.

  4. Olfactory cells via nasal biopsy reflect the developing brain in gene expression profiles: utility and limitation of the surrogate tissues in research for brain disorders.

    PubMed

    Horiuchi, Yasue; Kano, Shin-Ichi; Ishizuka, Koko; Cascella, Nicola G; Ishii, Seiji; Talbot, C Conover; Jaffe, Andrew E; Okano, Hideyuki; Pevsner, Jonathan; Colantuoni, Carlo; Sawa, Akira

    2013-12-01

    Human olfactory cells obtained by rapid nasal biopsy have been suggested to be a good surrogate system to address brain disease-associated molecular changes. Nonetheless, whether use of this experimental strategy is justified remains unclear. Here we compared expression profiles of olfactory cells systematically with those from the brain tissues and other cells. Principal component analysis indicated that the expression profiles of olfactory cells are very different from those of blood cells, but are closer to those of stem cells, in particular mesenchymal stem cells, that can be differentiated into the cells of the central nervous system.

  5. Olfactory cells via nasal biopsy reflect the developing brain in gene expression profiles: utility and limitation of the surrogate tissues in research for brain disorders

    PubMed Central

    Horiuchi, Yasue; Kano, Shin-ichi; Ishizuka, Koko; Cascella, Nicola G.; Ishii, Seiji; Talbot, C. Conover; Jaffe, Andrew E.; Okano, Hideyuki; Pevsner, Jonathan; Colantuoni, Carlo; Sawa, Akira

    2014-01-01

    Human olfactory cells obtained by rapid nasal biopsy have been suggested to be a good surrogate system to address brain disease-associated molecular changes. Nonetheless, whether use of this experimental strategy is justified remains unclear. Here we compared expression profiles of olfactory cells systematically with those from the brain tissues and other cells. Principal component analysis indicated that the expression profiles of olfactory cells are very different from those of blood cells, but are closer to those of stem cells, in particular mesenchymal stem cells, that can be differentiated into the cells of the central nervous system. PMID:24120685

  6. Expression of Galpha14 in sweet-transducing taste cells of the posterior tongue

    PubMed Central

    Tizzano, Marco; Dvoryanchikov, Gennady; Barrows, Jennell K; Kim, Soochong; Chaudhari, Nirupa; Finger, Thomas E

    2008-01-01

    Background "Type II"/Receptor cells express G protein-coupled receptors (GPCRs) for sweet, umami (T1Rs and mGluRs) or bitter (T2Rs), as well as the proteins for downstream signalling cascades. Transduction downstream of T1Rs and T2Rs relies on G-protein and PLCβ2-mediated release of stored Ca2+. Whereas Gαgus (gustducin) couples to the T2R (bitter) receptors, which Gα-subunit couples to the sweet (T1R2 + T1R3) receptor is presently not known. We utilized RT-PCR, immunocytochemistry and single-cell gene expression profiling to examine the expression of the Gαq family (q, 11, 14) in mouse taste buds. Results By RT-PCR, Gα14 is expressed strongly and in a taste selective manner in posterior (vallate and foliate), but not anterior (fungiform and palate) taste fields. Gαq and Gα11, although detectable, are not expressed in a taste-selective fashion. Further, expression of Gα14 mRNA is limited to Type II/Receptor cells in taste buds. Immunocytochemistry on vallate papillae using a broad Gαq family antiserum reveals specific staining only in Type II taste cells (i.e. those expressing TrpM5 and PLCβ2). This staining persists in Gαq knockout mice and immunostaining with a Gα11-specific antiserum shows no immunoreactivity in taste buds. Taken together, these data show that Gα14 is the dominant Gαq family member detected. Immunoreactivity for Gα14 strongly correlates with expression of T1R3, the taste receptor subunit present in taste cells responsive to either umami or sweet. Single cell gene expression profiling confirms a tight correlation between the expression of Gα14 and both T1R2 and T1R3, the receptor combination that forms sweet taste receptors. Conclusion Gα14 is co-expressed with the sweet taste receptor in posterior tongue, although not in anterior tongue. Thus, sweet taste transduction may rely on different downstream transduction elements in posterior and anterior taste fields. PMID:19014514

  7. 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.

  8. Phosphonate utilization by bacteria.

    PubMed Central

    Cook, A M; Daughton, C G; Alexander, M

    1978-01-01

    Bacteria able to use at least one of 13 ionic alkylphosphonates of O-alkyl or O,O-dialkyl alkylphosphonates as phosphorus sources were isolated from sewage and soil. Four of these isolates used 2-aminoethylphosphonic acid (AEP) as a sole carbon, nitrogen, and phosphorus source. None of the other phosphonates served as a carbon source for the organisms. One isolate, identified as Pseudomonas putida, grew with AEP as its sole carbon, nitrogen, and phosphorus source and released nearly all of the organic phosphorus as orthophosphate and 72% of the AEP nitrogen as ammonium. This is the first demonstration of utilization of a phosphonoalkyl moiety as a sole carbon source. Cell-free extracts of P. putida contained an inducible enzyme system that required pyruvate and pyridoxal phosphate to release orthophosphate from AEP; acetaldehyde was tentatively identified as a second product. Phosphite inhibited the enzyme system. PMID:618850

  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, 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...

  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, 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...

  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, 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...

  13. Enhancement of light harvesting efficiency of silicon solar cell utilizing arrays of poly(methyl methacrylate-co-acrylic acid) nano-spheres and nano-spheres with embedded silver nano-particles

    NASA Astrophysics Data System (ADS)

    Lee, Chee-Leong; Goh, Wee-Sheng; Chee, Swee-Yong; Yik, Lai-Kuan

    2017-02-01

    An array of uniformly distributed monolayer of poly(methyl methacrylate-co-acrylic acid) nano-spheres were deposited onto an amorphous silicon photovoltaic cell utilizing dip coating technique. The electrical characteristics of the coated photovoltaic cell reveal that the nano-spheres with an average diameter size of 101 nm exhibits excellent light harvesting characteristics if compared to the nano-spheres of other sizes. The power conversion efficiency from such integration of the nano-structures (i.e. 3.14% per PV cell) indicates that at least 1.6 times of improvement (or relative enhancement of 57%) can be achieved comparatively to the uncoated photovoltaic cell (i.e. 2% per PV cell). Further increment of the power conversion efficiency of the solar cell has been attained with the incorporation of the silver nano-particles into the nano-spheres of similar average size. With the inclusion of the silver nano-particles into such nano-spheres, the power conversion efficiency of the solar cell has attained 5.57% per PV cell, which is about 2.8 times (or relative enhancement of 179%) if compared to the uncoated samples. Hence, this novel and controllable technique of fabricating omnidirectional light-harvesting nano-spheres with embedded silver nano-particles will indubitably be beneficial to various types of optoelectronic devices.

  14. The Experienced Utility of Expected Utility Approaches

    DTIC Science & Technology

    1980-04-01

    than one using unit weights. Similarly, goods obtained with high probability should be valued more than those obtained with low probability. Therefore...consequences are added, low or negative utility associated with one conse- quence can, in principle, be compensated for by sufficiently high utility on...calibrated probability assessor should have more true statements associated with high than with low probabilities. Specifically, XX% of the statements

  15. Utilization and utility of immunohistochemistry in dermatopathology.

    PubMed

    Naert, Karen A; Trotter, Martin J

    2013-02-01

    Immunohistochemistry (IHC) is considered a valuable ancillary tool for dermatopathology diagnosis, but few studies have measured IHC utilization by dermatopathologists or assessed its diagnostic utility. In a regionalized, community-based dermatopathology practice, we measured IHC utilization (total requests, specific antibodies requested, and final diagnosis) over a 12-month period. Next, we assessed diagnostic utility by comparing a preliminary "pre-IHC" diagnosis based on routine histochemical staining with the final diagnosis rendered after consideration of IHC results. The dermatopathology IHC utilization rate was 1.2%, averaging 3.6 stains requested per case. Melanocytic, hematolymphoid, and fibrohistiocytic lesions made up 23%, 18%, and 16%, respectively, of the total cases requiring IHC. S100 and Melan A were the most frequently requested stains, ordered on 50% and 34% of IHC cases, respectively. The utility study revealed that IHC changed the diagnosis in 11%, confirmed a diagnosis, or excluded a differential diagnosis in 77%, and was noncontributory in 4% of cases. Where IHC results prompted a change in diagnosis, 14% were a change from a benign to malignant lesion, whereas 32% changed from one malignant entity to another. IHC is most commonly used in cutaneous melanocytic and hematolymphoid lesions. In 11% of dermatopathology cases in which IHC is used, information is provided that changes the H&E diagnosis. Such changes may have significant treatment implications. IHC is noncontributory in only a small percentage of cases.

  16. 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

  17. 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.

  18. 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.

  19. 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

  20. Utilities Expense Report.

    ERIC Educational Resources Information Center

    Moore, Deborah P.

    2001-01-01

    Examines how deregulation has affected school district utility costs. Offers ideas that can help school districts save money and energy. Provides several examples of state-wide initiatives intended to help school districts control utility costs. (GR)

  1. Alginate as a cell culture substrate for growth and differentiation of human retinal pigment epithelial cells.

    PubMed

    Heidari, Razeih; Soheili, Zahra-Soheila; Samiei, Shahram; Ahmadieh, Hamid; Davari, Maliheh; Nazemroaya, Fatemeh; Bagheri, Abouzar; Deezagi, Abdolkhalegh

    2015-03-01

    The purpose of this study was to evaluate retinal pigment epithelium (RPE) cells' behavior in alginate beads that establish 3D environment for cellular growth and mimic extracellular matrix versus the conventional 2D monolayer culture. RPE cells were encapsulated in alginate beads by dripping alginate cell suspension into CaCl2 solution. Beads were suspended in three different media including Dulbecco's modified Eagle's medium (DMEM)/F12 alone, DMEM/F12 supplemented with 10 % fetal bovine serum (FBS), and DMEM/F12 supplemented with 30 % human amniotic fluid (HAF). RPE cells were cultivated on polystyrene under the same conditions as controls. Cell phenotype, cell proliferation, cell death, and MTT assay, immunocytochemistry, and real-time RT-PCR were performed to evaluate the effect of alginate on RPE cells characteristics and integrity. RPE cells can survive and proliferate in alginate matrixes. Immunocytochemistry analysis exhibited Nestin, RPE65, and cytokeratin expressions in a reasonable number of cultured cells in alginate beads. Real-time PCR data demonstrated high levels of Nestin, CHX10, RPE65, and tyrosinase gene expressions in RPE cells immobilized in alginate when compared to 2D monolayer culture systems. The results suggest that alginate can be used as a reliable scaffold for maintenance of RPE cells' integrity and in vitro propagation of human retinal progenitor cells for cell replacement therapies in retinal diseases.

  2. Effects of Substrate Mechanics on Contractility of Cardiomyocytes Generated from Human Pluripotent Stem Cells

    PubMed Central

    Hazeltine, Laurie B.; Simmons, Chelsey S.; Salick, Max R.; Lian, Xiaojun; Badur, Mehmet G.; Han, Wenqing; Delgado, Stephanie M.; Wakatsuki, Tetsuro; Crone, Wendy C.; Pruitt, Beth L.; Palecek, Sean P.

    2012-01-01

    Human pluripotent stem cell (hPSC-) derived cardiomyocytes have potential applications in drug discovery, toxicity testing, developmental studies, and regenerative medicine. Before these cells can be reliably utilized, characterization of their functionality is required to establish their similarity to native cardiomyocytes. We tracked fluorescent beads embedded in 4.4–99.7 kPa polyacrylamide hydrogels beneath contracting neonatal rat cardiomyocytes and cardiomyocytes generated from hPSCs via growth-factor-induced directed differentiation to measure contractile output in response to changes in substrate mechanics. Contraction stress was determined using traction force microscopy, and morphology was characterized by immunocytochemistry for α-actinin and subsequent image analysis. We found that contraction stress of all types of cardiomyocytes increased with substrate stiffness. This effect was not linked to beating rate or morphology. We demonstrated that hPSC-derived cardiomyocyte contractility responded appropriately to isoprenaline and remained stable in culture over a period of 2 months. This study demonstrates that hPSC-derived cardiomyocytes have appropriate functional responses to substrate stiffness and to a pharmaceutical agent, which motivates their use in further applications such as drug evaluation and cardiac therapies. PMID:22649451

  3. 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)

  4. Sourcebook on Research Utilization.

    ERIC Educational Resources Information Center

    Rubin, Allen, Ed.; Rosenblatt, Aaron, Ed.

    Major papers presented at the Conference on Research Utilization in Social Work Education are compiled in this sourcebook. The conference focused on six topics that reviewed the state of the art of research utilization and suggested directions for the future. The papers included are: Understanding Research Utilization in Social Work (Stuart A.…

  5. 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.

  6. Clinical utility of neuronal cells directly converted from fibroblasts of patients for neuropsychiatric disorders: studies of lysosomal storage diseases and channelopathy

    PubMed Central

    Kano, Shin-ichi; Yuan, Ming; Cardarelli, Ross A.; Maegawa, Gustavo; Higurashi, Norimichi; Gaval-Cruz, Meriem; Wilson, Ashley M.; Tristan, Carlos; Kondo, Mari A.; Chen, Yian; Koga, Minori; Obie, Cassandra; Ishizuka, Koko; Seshadri, Saurav; Srivastava, Rupali; Kato, Takahiro A.; Horiuchi, Yasue; Sedlak, Thomas W.; Lee, Yohan; Rapoport, Judith L.; Hirose, Shinichi; Okano, Hideyuki; Valle, David; O'Donnell, Patricio; Sawa, Akira; Kai, Mihoko

    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

  7. An economical and highly productive cell-free protein synthesis system utilizing fructose-1,6-bisphosphate as an energy source.

    PubMed

    Kim, Tae-Wan; Keum, Jung-Won; Oh, In-Seok; Choi, Cha-Yong; Kim, Ho-Cheol; Kim, Dong-Myung

    2007-07-15

    In this study, we describe the development of a cost effective and highly productive cell-free protein synthesis system derived from Escherichia coli. Through the use of an optimal energy source and cell extract, approximately 1.3mg/mL of protein was generated from a single batch reaction at greatly reduced reagent costs. Compared to previously reported systems, the described method yields approximately 14-fold higher productivity per unit reagent cost making this cell-free synthesis technique a promising alternative for more efficient protein production.

  8. Utilizing two-photon fluorescence and second harmonic generation microscopy to study human bone marrow mesenchymal stem cell morphogenesis in chitosan scaffold

    NASA Astrophysics Data System (ADS)

    Su, Ping-Jung; Huang, Chi-Hsiu; Huang, Yi-You; Lee, Hsuan-Sue; Dong, Chen-Yuan

    2008-02-01

    A major goal of tissue engineering is to cultivate the cartilage in vitro. One approach is to implant the human bone marrow mesenchymal stem cells into the three dimensional biocompatible and biodegradable material. Through the action of the chondrogenic factor TGF-β3, the stem cells can be induced to secrete collagen. In this study, mesenchymal stem cells are implanted on the chitosan scaffold and TGF-β3 was added to produce the cartilage tissue and TP autofluorescence and SHG microscopy was used to image the process of chondrogenesis. With additional development, multiphoton microscopy can be developed into an effective tool for evaluating the quality of tissue engineering products.

  9. Utilization of photoinduced charge-separated state of donor-acceptor-linked molecules for regulation of cell membrane potential and ion transport.

    PubMed

    Numata, Tomohiro; Murakami, Tatsuya; Kawashima, Fumiaki; Morone, Nobuhiro; Heuser, John E; Takano, Yuta; Ohkubo, Kei; Fukuzumi, Shunichi; Mori, Yasuo; Imahori, Hiroshi

    2012-04-11

    The control of ion transport across cell membranes by light is an attractive strategy that allows targeted, fast control of precisely defined events in the biological membrane. Here we report a novel general strategy for the control of membrane potential and ion transport by using charge-separation molecules and light. Delivery of charge-separation molecules to the plasma membrane of PC12 cells by a membranous nanocarrier and subsequent light irradiation led to depolarization of the membrane potential as well as inhibition of the potassium ion flow across the membrane. Photoregulation of the cell membrane potential and ion transport by using charge-separation molecules is highly promising for control of cell functions.

  10. Ig heavy chain third complementarity determining regions (H CDR3s) after stem cell transplantation do not resemble the developing human fetal H CDR3s in size distribution and Ig gene utilization.

    PubMed

    Gokmen, E; Raaphorst, F M; Boldt, D H; Teale, J M

    1998-10-15

    Previous studies have suggested that the B-cell repertoire after stem cell transplantation resembles the developing repertoire in the fetus. Fetal and adult repertoires differ strikingly at the molecular level in Ig heavy chain third complementarity determining region (H CDR3) size distribution and Ig gene utilization. Previously, the posttransplant repertoire has not been studied fully in this regard. In this study, we analyzed H CDR3s posttransplant using CDR3 fingerprinting, single-strand conformation polymorphism (SSCP), and random sequencing. Eleven adult patients who received either autologous (n = 6) or allogeneic adult sibling (n = 5) hematopoietic stem cell transplants were studied. IgM H CDR3 repertoires demonstrated limited clonal diversity within the first 6 to 10 weeks posttransplant. By 3 to 4 months, the IgM H CDR3 repertoires were as diverse as those in healthy adults. Reconstitution of the IgM diversity correlated with the expansion of the multimember VH3 family. By contrast, the contribution of the single-member VH6 family was limited in most patients up to 6 to 9 months. No evidence was seen for greater contribution of VH6 posttransplant. IgG repertoires remained clonally restricted at all times. In all patients, H CDR3 sizes fell within adult limits. Direct nucleotide sequencing of H CDR3s showed adult-type N-nucleotide insertions and Ig gene utilization. These results indicate that the emerging repertoire posttransplant does not resemble the developing fetal repertoire at the molecular level.

  11. 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.

  12. Pilot Study of iPS-derived Neural Cells to Examine Biological Effects of Alcohol on Human Neurons in vitro

    PubMed Central

    Lieberman, Richard; Levine, Eric S.; Kranzler, Henry R.; Abreu, Christine; Covault, Jonathan

    2012-01-01

    Background Studies of the effects of alcohol on NMDA receptor function and gene expression have depended on rodent or post-mortem human brain models. Ideally, the effects of alcohol might better be examined in living neural tissue derived from human subjects. In this study, we used new technologies to reprogram human subject-specific tissue into pluripotent cell colonies and generate human neural cultures as a model system to examine the molecular actions of alcohol. Methods Induced pluripotent stem (iPS) cells were generated from skin biopsies taken from 7 individuals, 4 alcohol dependent subjects and 3 social drinkers.. We differentiated the iPS cells into neural cultures and characterized them by immunocytochemistry using antibodies for the neuronal marker beta III-tubulin, glial marker s100β, and synaptic marker synpasin1. Electrophysiology was performed to characterize the iPS-derived neurons and measure the effects of acute alcohol exposure on the NMDA receptor response in chronically alcohol exposed and non-exposed neural cultures from one non-alcoholic. Finally, we examined changes in mRNA expression of the NMDA receptor subunit genes GRIN1, GRIN2A, GRIN2B, and GRIN2D after 7 days of alcohol exposure and after 24-hour withdrawal from chronic alcohol exposure. Results Immunocytochemistry revealed positive staining for neuronal, glial, and synaptic markers. iPS-derived neurons displayed spontaneous electrical properties and functional ionotropic receptors. Acute alcohol exposure significantly attenuated the NMDA response, an effect that was not observed after 7 days of chronic alcohol exposure. After 7 days of chronic alcohol exposure, there were significant increases in mRNA expression of GRIN1, GRIN2A, and GRIN2D in cultures derived from alcoholic subjects but not in cultures derived from non-alcoholics. Conclusions These findings support the potential utility of human iPS-derived neural cultures as in vitro models to examine the molecular actions of

  13. 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.

  14. 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

    2016-01-05

    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.

  15. 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

  16. Utility Assessment Methods.

    DTIC Science & Technology

    1982-05-01

    Edwards, W., "Behavioral Decision Theory ," Ar -7. Review of Psychology, Vol. 12 (1961), pp. 473-498. 28. Edwards, W., "How to Use Multiattribute Utility ...Economics and Management Science, Northwestern University, Evanston, Illinois (1981). 31. Farquhar, P.H., "A Survey of Multiattribute Utility Theory and...89. -43- 32. Farquhar, P.H., "Advances in Multiattribute Utility Theory ," Theory and Decision, Vol. 12 (1980), pp. 381-394. 33. Fischer, G.W

  17. 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.

  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.

  19. Underestimating the safety benefits of a new vaccine: the impact of acellular pertussis vaccine versus whole-cell pertussis vaccine on health services utilization.

    PubMed

    Hawken, Steven; Manuel, Douglas G; Deeks, Shelley L; Kwong, Jeffrey C; Crowcroft, Natasha S; Wilson, Kumanan

    2012-12-01

    The population-level safety benefits of the acellular pertussis vaccine may have been underestimated because only specific adverse events were considered, not overall impact on health services utilization. Using the Vaccine and Immunization Surveillance in Ontario (VISION) system, the authors analyzed data on 567,378 children born between April 1994 and March 1996 (before introduction of acellular pertussis vaccine) and between April 1998 and March 2000 (after introduction of acellular pertussis vaccine) in Ontario, Canada. Using the self-controlled case series study design, they examined emergency room visits and hospital admissions occurring after routine pediatric vaccinations. The authors determined the relative incidence of events taking place before introduction of the acellular vaccine versus after introduction by calculating relative incidence ratios (RIRs). The observed RIRs demonstrated a highly statistically significant reduction in relative incidence after introduction of the acellular vaccine. RIRs for vaccine administered at ages 2, 4, 6, and 18 months were 1.82 (95% confidence interval (CI): 1.64, 2.01), 1.91 (95% CI: 1.71, 2.13), 1.54 (95% CI: 1.38, 1.72), and 1.51 (95% CI: 1.34, 1.69), respectively, comparing event rates before the introduction of acellular vaccine with those after introduction. The authors estimated that approximately 90 emergency room visits and 9 admissions per month were avoided by switching to the acellular vaccine, which is a 38-fold higher impact than when they considered only admissions for febrile and afebrile convulsions. Future analyses comparing vaccines for safety should examine specific endpoints and general health services utilization.

  20. Expression of cholecystokinin2-receptor in rat and human L cells and the stimulation of glucagon-like peptide-1 secretion by gastrin treatment.

    PubMed

    Cao, Yang; Cao, Xun; Liu, Xiao-Min

    2015-03-01

    Gastrin is a gastrointestinal hormone secreted by G cells. Hypergastrinemia can improve blood glucose and glycosylated hemoglobin levels. These positive effects are primarily due to the trophic effects of gastrin on β-cells. In recent years, many receptors that regulate secretion of glucagon-like peptide 1 (GLP-1) have been identified in enteroendocrine L cell lines. This led us to hypothesize that, in addition to the trophic effects of gastrin on β-cells, L cells also express cholecystokinin2-receptor (CCK2R), which may regulate GLP-1 secretion and have synergistic effects on glucose homeostasis. Our research provides a preliminary analysis of CCK2R expression and the stimulating effect of gastrin treatment on GLP-1 secretion in a human endocrine L cell line, using RT-PCR, Western blot, immunocytochemistry, and ELISA analyses. The expression of proglucagon and prohormone convertase 3, which regulate GLP-1 biosynthesis, were also analyzed by real-time PCR. Double immunofluorescence labeling was utilized to assess the intracellular localization of CCK2R and GLP-1 in L cells harvested from rat colon tissue. Our results showed that CCK2R was expressed in both the human L cell line and the rat L cells. We also showed that treatment with gastrin, a CCK2R agonist, stimulated the secretion of GLP-1, and that this effect was likely due to increased expression of proglucagon and PCSK1 (also known as prohormone convertase 3 (PC3 gene)). These results not only provide a basis for the role gastrin may play in intestinal L cells, and may also provide the basis for the development of a method of gastrin-mediated glycemic regulation.

  1. THE EVALUATION OF PEPTIDE/HISTIDINE TRANSPORTER 1 (PHT1) FUNCTION: UPTAKE KINETICS UTILIZING A COS-7 STABLY TRANSFECTED CELL LINE.

    PubMed

    Lindley, David J; Carl, Stephen M; Mowery, Stephanie A; Knipp, Gregory T

    2011-10-01

    There have been relatively few studies focused on the proton-dependent oligopeptide transporter (POT) superfamily member, Peptide/Histidine Transporter 1 (PHT1), with respect to its contribution to the ADME of peptides and peptide-based drugs. These studies were conducted to determine hPHT1-mediated, H(+)-dependent uptake kinetics of histidine, carnosine, Gly-Sar and valacyclovir in stably transfected hPHT1-COS-7 cells comparative to kinetics determined in an empty vector (Mock) stably transfected cell line. The results suggest that Gly-Sar appears to be a substrate for PHT1 based on efflux from the stably transfected hPHT1 COS-7 cells. Histidine and Gly-Sar concentration- and time-dependent studies suggest mixed-uptake kinetics. These studies suggest that stably transfected hPHT1-COS-7 cells exhibit different uptake kinetics than those observed in our previous studies and illustrate the requirement for experiments to delineate the physiological role of hPHT1.

  2. 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

  3. 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.

  4. Bright fluorescence monitoring system utilizing Zoanthus sp. green fluorescent protein (ZsGreen) for human G-protein-coupled receptor signaling in microbial yeast cells.

    PubMed

    Nakamura, Yasuyuki; Ishii, Jun; Kondo, Akihiko

    2013-01-01

    G-protein-coupled receptors (GPCRs) are currently the most important pharmaceutical targets for drug discovery because they regulate a wide variety of physiological processes. Consequently, simple and convenient detection systems for ligands that regulate the function of GPCR have attracted attention as powerful tools for new drug development. We previously developed a yeast-based fluorescence reporter ligand detection system using flow cytometry. However, using this conventional detection system, fluorescence from a cell expressing GFP and responding to a ligand is weak, making detection of these cells by fluorescence microscopy difficult. We here report improvements to the conventional yeast fluorescence reporter assay system resulting in the development of a new highly-sensitive fluorescence reporter assay system with extremely bright fluorescence and high signal-to-noise (S/N) ratio. This new system allowed the easy detection of GPCR signaling in yeast using fluorescence microscopy. Somatostatin receptor and neurotensin receptor (implicated in Alzheimer's disease and Parkinson's disease, respectively) were chosen as human GPCR(s). The facile detection of binding to these receptors by cognate peptide ligands was demonstrated. In addition, we established a highly sensitive ligand detection system using yeast cell surface display technology that is applicable to peptide screening, and demonstrate that the display of various peptide analogs of neurotensin can activate signaling through the neurotensin receptor in yeast cells. Our system could be useful for identifying lead peptides with agonistic activity towards targeted human GPCR(s).

  5. 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

  6. 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.

  7. Exosome-based tumor antigens-adjuvant co-delivery utilizing genetically engineered tumor cell-derived exosomes with immunostimulatory CpG DNA.

    PubMed

    Morishita, Masaki; Takahashi, Yuki; Matsumoto, Akihiro; Nishikawa, Makiya; Takakura, Yoshinobu

    2016-12-01

    For cancer immunotherapy via tumor antigen vaccination in combination with an adjuvant, major challenges include the identification of a particular tumor antigen and efficient delivery of the antigen as well as adjuvant to antigen-presenting cells. In this study, we proposed an efficient exosome-based tumor antigens-adjuvant co-delivery system using genetically engineered tumor cell-derived exosomes containing endogenous tumor antigens and immunostimulatory CpG DNA. Murine melanoma B16BL6 cells were transfected with a plasmid vector encoding a fusion streptavidin (SAV; a protein that binds to biotin with high affinity)-lactadherin (LA; an exosome-tropic protein) protein, yielding genetically engineered SAV-LA-expressing exosomes (SAV-exo). SAV-exo were combined with biotinylated CpG DNA to prepare CpG DNA-modified exosomes (CpG-SAV-exo). Fluorescent microscopic observation revealed the successful modification of exosomes with CpG DNA by SAV-biotin interaction. CpG-SAV-exo showed efficient and simultaneous delivery of exosomes with CpG DNA to murine dendritic DC2.4 cells in culture. Treatment with CpG-SAV-exo effectively activated DC2.4 cells and enhanced tumor antigen presentation capacity. Immunization with CpG-SAV-exo exhibited stronger in vivo antitumor effects in B16BL6 tumor-bearing mice than simple co-administration of exosomes and CpG DNA. Thus, genetically engineered CpG-SAV-exo is an effective exosome-based tumor antigens-adjuvant co-delivery system that will be useful for cancer immunotherapy.

  8. CD4(+)CD25(+) T regulatory cells inhibit CD8(+) IFN-gamma production during acute and chronic FIV infection utilizing a membrane TGF-beta-dependent mechanism.

    PubMed

    Fogle, Jonathan E; Mexas, Angela M; Tompkins, Wayne A; Tompkins, Mary B

    2010-02-01

    CD8(+) lymphocytes are critical to the control and elimination of viral pathogens. Impaired CD8(+) responses are well recognized in lentiviral infections; however, the mechanisms underlying CD8(+) impairment remain elusive. Using the feline immunodeficiency virus (FIV) model for human AIDS, we reported previously that CD4(+)CD25(+) Treg cells in both the acute and long-term, asymptomatic phase of infection are constitutively activated and suppress CD4(+)CD25(-) T cell responses. In the current study, we have demonstrated that CD4(+)CD25(+) Treg cells suppress CD8(+) responses to immune stimulation during both the acute and chronic, asymptomatic phase of FIV infection and that the mechanism of suppression may be mediated by membrane-associated TGF-beta (mTGF-beta) on CD4(+)CD25(+) lymphocytes. Depletion of CD4(+)CD25(+) lymphocytes from lymph node suspensions significantly enhanced production of IFN-gamma during the acute phase of infection and coculture of CD8(+) lymphocytes with CD4(+)CD25(+) lymphocytes resulted in suppression of CD8(+) IFN-gamma during both the acute and chronic stages of infection. FACS analysis indicated that there was TGF-betaRII upregulation on CD8(+) cells from FIV(+) cats during the acute and chronic stage of infection. In addition, there was upregulation of mTGF-beta on the CD4(+)CD25(+) subset in chronically infected cats. In support of activation of the TGF-beta signaling pathway, Western blotting showed Smad 2 phosphorylation in CD8(+) targets following CD4(+)CD25(+)/CD8(+) coculture. These results demonstrate the suppressive effect CD4(+)CD25(+) Treg cells have on the CD8(+) immune response during the acute and chronic stages of FIV infection and suggest that the mechanism of suppression may be mediated by mTGF-beta.

  9. CD4+CD25+ T Regulatory Cells Inhibit CD8+ IFN-γ Production During Acute and Chronic FIV Infection Utilizing a Membrane TGF-β-Dependent Mechanism

    PubMed Central

    Fogle, Jonathan E.; Mexas, Angela M.; Tompkins, Wayne A.

    2010-01-01

    Abstract CD8+ lymphocytes are critical to the control and elimination of viral pathogens. Impaired CD8+ responses are well recognized in lentiviral infections; however, the mechanisms underlying CD8+ impairment remain elusive. Using the feline immunodeficiency virus (FIV) model for human AIDS, we reported previously that CD4+CD25+ Treg cells in both the acute and long-term, asymptomatic phase of infection are constitutively activated and suppress CD4+CD25– T cell responses. In the current study, we have demonstrated that CD4+CD25+ Treg cells suppress CD8+ responses to immune stimulation during both the acute and chronic, asymptomatic phase of FIV infection and that the mechanism of suppression may be mediated by membrane-associated TGF-β (mTGF-β) on CD4+CD25+ lymphocytes. Depletion of CD4+CD25+ lymphocytes from lymph node suspensions significantly enhanced production of IFN-γ during the acute phase of infection and coculture of CD8+ lymphocytes with CD4+CD25+ lymphocytes resulted in suppression of CD8+ IFN-γ during both the acute and chronic stages of infection. FACS analysis indicated that there was TGF-βRII upregulation on CD8+ cells from FIV+ cats during the acute and chronic stage of infection. In addition, there was upregulation of mTGF-β on the CD4+CD25+ subset in chronically infected cats. In support of activation of the TGF-β signaling pathway, Western blotting showed Smad 2 phosphorylation in CD8+ targets following CD4+CD25+/CD8+ coculture. These results demonstrate the suppressive effect CD4+CD25+ Treg cells have on the CD8+ immune response during the acute and chronic stages of FIV infection and suggest that the mechanism of suppression may be mediated by mTGF-β. PMID:20156102

  10. International utilization and operations

    NASA Technical Reports Server (NTRS)

    Goldberg, Stanley R.

    1989-01-01

    The international framework of the Space Station Freedom Program is described. The discussion covers the U.S. space policy, international agreements, international Station elements, overall program management structure, and utilization and operations management. Consideration is also given to Freedom's user community, Freedom's crew, pressurized payload and attached payload accommodations, utilization and operations planning, user integration, and user operations.

  11. Utilization Research. Chapter 11.

    ERIC Educational Resources Information Center

    1996

    This collection of papers presented at a 1996 conference on children's mental health focuses on utilization research. Papers have the following titles and authors: (1) "Information Equity: A Critical Component of Strong Service Systems" (Catherine Batsche and Allison Metcalf); (2) "Utilization of Children's Mental Health Services: Differentiating…

  12. 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).

  13. Prostaglandin E2 activates and utilizes mTORC2 as a central signaling locus for the regulation of mast cell chemotaxis and mediator release.

    PubMed

    Kuehn, Hye Sun; Jung, Mi-Yeon; Beaven, Michael A; Metcalfe, Dean D; Gilfillan, Alasdair M

    2011-01-07

    Prostaglandin (PG) E(2), a potent mediator produced in inflamed tissues, can substantially influence mast cell responses including adhesion to basement membrane proteins, chemotaxis, and chemokine production. However, the signaling pathways by which PGE(2) induces mast cell chemotaxis and chemokine production remains undefined. In this study, we identified the downstream target of phosphatidylinositol 3-kinase, mammalian target of rapamycin (mTOR), as a key regulator of these responses. In mouse bone marrow-derived mast cells, PGE(2) was found to induce activation of mTORC1 (mTOR complexed to raptor) as indicated by increased p70S6K and 4E-BP1 phosphorylation, and activation of mTORC2 (mTOR complexed to rictor), as indicated by increased phosphorylation of AKT at position Ser(473). Selective inhibition of the mTORC1 cascade by rapamycin or by the use of raptor-targeted shRNA failed to decrease PGE(2)-mediated chemotaxis or chemokine generation. However, inhibition of the mTORC2 cascade through the dual mTORC1/mTORC2 inhibitor Torin, or through rictor-targeted shRNA, resulted in a significant attenuation in PGE(2)-mediated chemotaxis, which was associated with a comparable decrease in actin polymerization. Furthermore, mTORC2 down-regulation decreased PGE(2)-induced production of the chemokine monocyte chemoattractant protein-1 (CCL2), which was linked to a significant reduction in ROS production. These findings are consistent with the conclusion that activation of mTORC2, downstream of PI3K, represents a critical signaling locus for chemotaxis and chemokine release from PGE(2)-activated mast cells.

  14. 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.

  15. Evaluation and refinement of a field-portable drinking water toxicity sensor utilizing electric cell-substrate impedance sensing and a fluidic biochip.

    PubMed

    Widder, Mark W; Brennan, Linda M; Hanft, Elizabeth A; Schrock, Mary E; James, Ryan R; van der Schalie, William H

    2015-07-01

    The US Army's need for a reliable and field-portable drinking water toxicity sensor was the catalyst for the development and evaluation of an electric cell-substrate impedance sensing (ECIS) device. Water testing technologies currently available to soldiers in the field are analyte-specific and have limited capabilities to detect broad-based water toxicity. The ECIS sensor described here uses rainbow trout gill epithelial cells seeded on fluidic biochips to measure changes in impedance for the detection of possible chemical contamination of drinking water supplies. Chemicals selected for testing were chosen as representatives of a broad spectrum of toxic industrial compounds. Results of a US Environmental Protection Agency (USEPA)-sponsored evaluation of the field portable device were similar to previously published US Army testing results of a laboratory-based version of the same technology. Twelve of the 18 chemicals tested following USEPA Technology Testing and Evaluation Program procedures were detected by the ECIS sensor within 1 h at USEPA-derived human lethal concentrations. To simplify field-testing methods further, elimination of a procedural step that acclimated cells to serum-free media streamlined the test process with only a slight loss of chemical sensitivity. For field use, the ECIS sensor will be used in conjunction with an enzyme-based sensor that is responsive to carbamate and organophosphorus pesticides.

  16. Isolation of murine postnatal brain microglia for phenotypic characterization using magnetic cell separation technology.

    PubMed

    Harms, Ashley S; Tansey, Malú G

    2013-01-01

    To shorten the time between brain harvesting and microglia isolation, and characterization, we utilized the MACS(®) neural dissociation kit followed by OctoMACS(®) CD11b magnetic bead isolation technique to positively select for brain microglia expressing the pan-microglial marker CD11b, a key subunit of the membrane attack complex (MAC). This protocol yields a viable and highly pure (>95%) microglial population of approximately 500,000 cells per pup that is amenable for in vitro characterization within hours or days after being harvested from brain tissue. Primary microglia from C57Bl/6 mice were plated for next-day analyses of morphology and cellular markers by immunocytochemistry or for analysis of gene expression under resting or LPS-stimulated conditions. The ease of isolation enables investigators to perform molecular and cellular analyses without having to wait 1-2 weeks to isolate microglia by conventional methods involving mechanical agitation to dislodge these from astrocyte beds.

  17. 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.

  18. 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.

  19. 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.

  20. Construction of three-dimensional dermo-epidermal skin equivalents using cell coating technology and their utilization as alternative skin for permeation studies and skin irritation tests.

    PubMed

    Akagi, Takami; Nagura, Mayuka; Hiura, Ayami; Kojima, Hajime; Akashi, Mitsuru

    2017-03-23

    In vitro generated human skin equivalents are generating interest as promising tools in basic research, as alternatives to animal testing and for clinical applications in regenerative medicine. For prediction of skin irritation and corrosion, three-dimensional (3D) human skin equivalents consisting of differentiated human keratinocytes have been developed and some models have been internationally accepted. However, more delicate assessments using full-thickness skin models, such as skin sensitization tests cannot be performed because of the lack of a dermis containing fibroblasts or appendages. In a previous study, we developed dermo-epidermal human skin equivalents (DESEs) using a cell coating technique, which employs cell surface coating by layer-by-layer (LbL) assembled extracellular matrix (ECM) films. The DESEs with dermis consisting of normal human dermal fibroblasts (NHDF) and epidermis consisting of human keratinocytes (KC) were easily fabricated by using this technology. In this study, the constructed DESEs were evaluated as an alternative skin for skin permeation and irritation tests. A good relationship of permeability coefficient of chemicals was observed between the DESEs and human skin data. We investigated whether the DESEs, a new in vitro skin model, are able to identify skin irritant and non-irritant substances among 20 reference chemicals. It was confirmed that the DESEs are applicable to skin irritation testing as defined in the European Centre for the Validation of Alternative Methods (ECVAM) Performance Standard (OECD Test Guideline 439). We further studied the construction of the DESEs with density-controlled blood capillary networks using human umbilical vein endothelial cells (HUVEC). The results suggest that DESEs allowing incorporation of skin appendages are more promising alternatives to animal testing, and can be applied to the design of physiologically relevant in vitro skin models.

  1. 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.

  2. TGF-beta and vitamin D3 utilize distinct pathways to suppress IL-12 production and modulate rapid differentiation of human monocytes into CD83+ dendritic cells.

    PubMed

    Lyakh, Lyudmila A; Sanford, Michael; Chekol, Sebel; Young, Howard A; Roberts, Anita B

    2005-02-15

    We previously demonstrated that agents known to signal infection or inflammation can rapidly and directly drive differentiation of human CD14+ monocytes into CD83+ dendritic cells (DCs) when introduced to cells under serum-free conditions. In this study, we evaluated the effects of TGF-beta and vitamin D3 (VitD3) on the proportion and function of monocytes that adopt DC characteristics. TGF-beta significantly decreased the proportion of cells that rapidly adopted stable DC characteristics in response to LPS, but had little or no effect on calcium ionophore-induced differentiation. In contrast, VitD3 showed no such pathway specificity and dramatically suppressed differentiation of monocytes into DCs in response to these agents. Both TGF-beta and VitD3 altered cytokine and chemokine production in LPS-treated monocytes, inhibited IL-12 and IL-10 secretion, and decreased the functional capacity of DCs. Despite the similar effects of TGF-beta and VitD3, there are significant differences in the signaling pathways used by these agents, as evidenced by their distinct effects on LPS- and calcium ionophore-induced DC differentiation, on LPS-induced secretion of IL-10, and on two members of the NF-kappaB family of transcription factors, RelB and cRel. These studies identify TGF-beta and VitD3 as potent regulatory factors that use distinct pathways to suppress both the differentiation of DCs as well as their capacity to secrete the Th1-polarizing cytokine IL-12. Because these agents are present in serum and negatively affect DC differentiation at physiological concentrations, our findings are likely to have significance regarding the in vivo role of TGF-beta and VitD3 in determining the type of immune responses.

  3. Increased glucose utilization and cell growth of Corynebacterium glutamicum by modifying the glucose-specific phosphotransferase system (PTS(Glc)) genes.

    PubMed

    Xu, Jianzhong; Zhang, Junlan; Liu, Dongdong; Zhang, Weiguo

    2016-12-01

    The phosphoenolpyruvate:glucose phosphotransferase system (PTS(Glc)) is the major pathway of glucose uptake in Corynebacterium glutamicum. This study investigated glucose consumption rate, cell growth, and metabolite changes resulting from modification of PTS(Glc). The classical l-lysine producer C. glutamicum XQ-8 exhibited low glucose consumption, cell growth, and l-lysine production rates, whereas these parameters were significantly increased during cultivating on glucose plus maltose, through inactivation of SugR, or by overexpression of PTS(Glc) genes. XQ-8sugR::cat/pDXW-8-ptsI exhibited the highest increase in glucose consumption, growth rate, and l-lysine production, followed by XQ-8sugR::cat/pDXW-8-ptsG. However, overexpression of ptsH had little effect on the above-mentioned factors. Although co-overexpression of ptsGHI led to the highest glucose consumption, growth rate, and final l-lysine production; the l-lysine production rate was lower than that of XQ-8sugR::cat/pDXW-8-ptsIH. In fed-batch fermentation, XQ-8sugR::cat/pDXW-8-ptsIH had a higher growth rate of 0.54 h(-1) to a dry cell mass of 66 g·L(-1) after 16 h, and had a higher l-lysine production rate of 159.2 g·L(-1) after 36 h. These results indicate that modification of the sugar transport systems improves amino acid production, especially for mutants obtained by repeated physical and (or) chemical mutagenesis. However, modification of these systems needs to be performed on a case-by-case basis.

  4. The prognostic utility and the association of serum light chains (free and total) and absolute lymphocyte count in patients with newly diagnosed diffuse large B-cell lymphoma.

    PubMed

    Han, Xiaohong; Wang, Jianfei; Zhang, Ningning; Yao, Jiarui; Feng, Yun; Li, Dan; Liu, Peng; Yang, Jianliang; Zhou, Shengyu; Qin, Yan; Yang, Sheng; Gui, Lin; He, Xiaohui; Shi, Yuankai

    2014-11-01

    In this study, serum free and total light chains (sFLC/sTLC) were measured in 108 serum samples of therapy-naïve patients with DLBCL. Clinicopathologic data and survival outcomes were analyzed according to the results of sFLC/sTLC measurements. Moreover, the association of sFLC/sTLC with absolute monocyte count (AMC) and absolute lymphocyte count (ALC) was evaluated. Elevated sFLC and abnormal κ/λ ratio was present in 42.6% (51/108) and 4.6% (5/108) of patients, respectively. sTLC was successfully measured in 107 serum samples, abnormal sTLC and abnormal κ/λ ratio was found in 28.0% (30/107) and 26.2% (28/107) of patients, respectively. Patients with elevated sFLC more frequently displayed adverse clinical characteristics, including age (P=0.001), B symptoms (P=0.022), low ALC (P=0.024) and hyperglobulinemia (P=0.012). Patients with elevated sFLC had an inferior overall survival (OS) (P=0.012) and tended to have shorter progression-free survival (PFS) (P=0.061) compared to patients with normal sFLC. Abnormal sTLC or abnormal sTLC ratio showed no significant association with clinical outcomes, with exception of abnormal concurrent κ and λ. Only association of sFLC and ALC with survival remained significant after adjusting for the International Prognostic Index (IPI). The measurement of sFLC and ALC at diagnosis might be useful for the prognostic stratification of patients and sTLC measurement was of little prognostic utility in DLBCL.

  5. Agreement between assays of cell-mediated immunity utilizing Mycobacterium bovis-specific antigens for the diagnosis of tuberculosis in African buffaloes (Syncerus caffer).

    PubMed

    Goosen, Wynand J; Miller, Michele A; Chegou, Novel N; Cooper,