27 resultados para elutriation


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Elutriation, as a means of sorting mineral particles, has received marked attention during the last fifteen years. Its use in the ceramics industry for the sorting of clays was recognized even before this.

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Intestinal intraepithelial lymphocytes (IEL) are specialized subsets of T cells with distinct functional capacities. While some IEL subsets are circulating, others such as CD8alphaalpha TCRalphabeta IEL are believed to represent non-circulating resident T cell subsets [Sim, G.K., Intraepithelial lymphocytes and the immune system. Adv. Immunol., 1995. 58: 297-343.]. Current methods to obtain enriched preparations of intraepithelial lymphocytes are mostly based on Percoll density gradient or magnetic bead-based technologies [Lundqvist, C., et al., Isolation of functionally active intraepithelial lymphocytes and enterocytes from human small and large intestine. J. Immunol. Methods, 1992. 152(2): 253-263.]. However, these techniques are hampered by a generally low yield of isolated cells, and potential artifacts due to the interference of the isolation procedure with subsequent functional assays, in particular, when antibodies against cell surface markers are required. Here we describe a new method for obtaining relatively pure populations of intestinal IEL (55-75%) at a high yield (>85%) by elutriation centrifugation. This technique is equally suited for the isolation and enrichment of intraepithelial lymphocytes of both mouse and human origin. Time requirements for fractionating cell suspensions by elutriation centrifugation are comparable to Percoll-, or MACS-based isolation procedures. Hence, the substantially higher yield and the consistent robust enrichment for intraepithelial lymphocytes, together with the gentle treatment of the cells during elutriation that does not interfere with subsequent functional assays, are important aspects that are in favor of using this elegant technology to obtain unmanipulated, unbiased populations of intestinal intraepithelial lymphocytes, and, if desired, also of pure epithelial cells.

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The method of isolation of bone marrow (BM) mesenchymal stem/stromal cells (MSCs) is a limiting factor in their study and therapeutic use. MSCs are typically expanded from BM cells selected on the basis of their adherence to plastic, which results in a heterogeneous population of cells. Prospective identification of the antigenic profile of the MSC population(s) in BM that gives rise to cells with MSC activity in vitro would allow the preparation of very pure populations of MSCs for research or clinical use. To address this issue, we used polychromatic flow cytometry and counterflow centrifugal elutriation to identify a phenotypically distinct population of mesenchymal stem/progenitor cells (MSPCs) within human BM. The MSPC activity resided within a population of rare, small CD45⁻CD73⁺CD90⁺CD105⁺ cells that lack CD44, an antigen that is highly expressed on culture-expanded MSCs. In culture, these MSPCs adhere to plastic, rapidly proliferate, and acquire CD44 expression. They form colony forming units-fibroblast and are able to differentiate into osteoblasts, chondrocytes, and adipocytes under defined in vitro conditions. Their acquired expression of CD44 can be partially downregulated by treatment with recombinant human granulocyte-colony stimulating factor, a response not found in BM-MSCs derived from conventional plastic adherence methods. These observations indicate that MSPCs within human BM are rare, small CD45⁻CD73⁺CD90⁺CD105⁺ cells that lack expression of CD44. These MSPCs give rise to MSCs that have phenotypic and functional properties that are distinct from those of BM-MSCs purified by plastic adherence.

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Although requirement for follicle stimulating hormone (FSH) in the initiation of spermatogenesis is well documented, its role in adult spermatogenesis is still debated. In the present communication, we have investigated the effect of specific immunoneutralization of FSH on apoptotic cell death in the testicular germ cells both in immature and adult rats. The germ cells of control animals showed predominantly high molecular weight DNA while the antiserum (a/s) treated group showed DNA fragmentation characteristic of apoptosis. The pattern could be detected within 24 hours of a/s treatment, and became more pronounced after 48 hours. The germ cells were purified from FSH a/s treated rats by centrifugal elutriation and vulnerability of each cell type to undergo apoptosis on FSH neutralization was investigated. The pachytene spermatocytes were found to be most sensitive to absence of FSH, even in the adult animals suggesting the involvement of FSH in spermatogenesis. The in situ analysis of DNA strand breakage following FSH a/s treatment showed fragmentation of the DNA of the pachytene spermatocytes confirming this observation. The in situ analysis also showed that the spermatogonia undergo apoptosis in addition to the pachytene spermatocytes. These data clearly demonstrate the role of FSH in the adult rat spermatogenesis.

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The selective withdrawal of pituitary gonadotropins through specific antibodies is known to cause disruption of spermatogenesis. The cellular mechanism responsible for the degenerative changes under isolated effect of luteinizing hormone (LH) deprivation is not clear. Using antibodies specific to LH we have investigated the effect of immunoneutralization of LH on apoptotic cell death in the testicular cells of the immature and the adult rats. Specific neutralization of LH resulted in apoptotic cell death of germ cells, both in the immature and the adult rats. The germ cells from control animals showed predominantly high molecular weight DNA, while the antiserum treated group showed DNA cleavage into low molecular weight DNA ladder characteristic of apoptosis. This pattern could be observed within 24 h of a/s administration and the effect could be reversed by testosterone. The germ cells were purified by centrifugal elutriation and the vulnerability of germ cell types to undergo apoptosis under LH deprivation was investigated. The round spermatids and the pachytene spermatocytes were found to be the most sensitive germ cells to lack of LH and underwent apoptosis. Interestingly, spermatogonial cells were found to be the least sensitive germ cells to the lack of LH in terms of apoptotic cell death. Results show that LH, in addition to being involved in the germ cell differentiation, is also involved in cell survival and prevent degeneration of germ cells during spermatogenesis. Apoptotic DNA fragmentation may serve as a useful marker for the study of hormonal regulation of spermatogenesis and the specific neutralization of gonadotropic hormones can be a reliable model for the study of the molecular mechanism of apoptosis.

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The present investigation was undertaken to establish a reference situation for future use, to identify temporal and spatial composition of macrofauna and estimate some ecological indices in the sub tidal waters along the Bushehr coastal waters in Persian Gulf. Six transects were selected including Genaveh, Farakeh, Shif, Bandargah, Rostami and Asalouyeh, at each transect 3 station were sampled in depths of zero, 5 and 10 metres. Sampling was seasonally carried out by a VAN VEEN grab 0.0225 m2, during summer 2008 until spring 2009. Samples were wet sieved immediately using 0.5 mm mesh size sieves and sediment retained in the sieve was preserved in 4% buffered formalin solution. Macrofauna specimen were separated from the sediments using decantation and elutriation methods, enumerated and identified up to the Genus level. Environmental factors such as temperature. pH, and salinity were recorded in field using sensitive probs and refractometer (for salinity) and also sediment samples were taken for TOM and grain size analysis in all the stations. 5611 specimens belonging to 66 genera were collected during the present study. Polychaetes were dominant both in terms of genus number (31) and relative abundance (74 % of total macrofaunal abundance). The other dominant groups were Artheropoda, (16.1%), Molusca (2.8%), Echinodermata (1.29%) and others including Nematoda, Nemertina, Echiura and Turbellaria (5.8%). Thirty one Genera belong of 27 families of polychaeta, one genus and family of Subphylum Chlicerata,19 genera belong to 14 families of Crustacea, 8 genera belong to 6 families of Molusca, were indentified in the studied region. 1 family (Polygordidae) and 3 genera (Flabeligera, Pilargis and Polygordius) of Polychaeta, 1 family (Nymphonidae) and genus (Nymphon) of Chelicerata, 1 Family (Nematoplanidae) and genus (Nematoplana) of Turbellaria, were identified for the first time in Persian Gulf area. The result indicated that macrofauna organism have strong relationship with the grain size characteristics of the sediments they inhabit. The most surface deposit feeder specimens such as Prionospio and Cossura were found in zero meters depth of Genaveh, Farakeh, Bandargah, Rostami and Asalouyeh stations with sandy substratum, however the most burrowing deposit feeder and scavenger specimens such as Capitella and Petaloproctus were collected in 5 and 10 meter depths of stations with silty–clay substratum. The annual mean abundance, Shanon- weiner diversity and evenness of macrofauna were estimated1152.73 N/ m² , 2.72 and 0.792 respectively .The annual average biomass and secondary production were computed 1.797 gDW m² and 3.594 gDW m² y-1 .The average of water temperature, salinity, pH and oxygen concentration were recorded between 16.37-36.05 °C, 38-42 g/l, 7.89-8.76 and 4.23-8.23 mg/l, respectively during this study in 6 studied region. Among of investigated stations Asalouyeh adjacent of effluent canal of Gas and petrochemical industry sewage and Farakeh regions adjacent the Helleh estuary had the lowets and the highest community indices. The average of diversity and density in 5 meters depth stations with moderate of sand, silt and clay were slightly more than 2 other depths stations, it seems that 5 meters stations are made a transition habitats between 2 sandy and clay habitats, that can be used by 2 groups of surface and borrowing deposit feeders. Based on the data provided in this survey, the temperature variation, sediment texture, TOM, type habitat and manmade factors of Gas and petrochemical industries have had the most effect on the macrofauna community structure in the studied region during sampling periods.

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The attrition of two potential oxygen-carriers for chemical-looping, 100. wt% mechanically-mixed, unsupported iron oxide (400-600 μm diameter) and 25. wt% copper oxide impregnated on alumina (600-900 μm diameter), has been studied. The rates of attrition of batches of these particles whilst they were being fluidised and subjected to successive cycles of reduction and oxidation were determined by measuring the rate of production of fine particles elutriated from the bed, as well as progressive changes in the distribution of particle sizes retained in the bed. The ability of the particles to withstand impacts was also investigated by examining the degree of fragmentation of 1. g of reacted particles of known size on projecting them at a target at various velocities. It was found that the mechanical strength of the iron oxide particles deteriorated significantly after repeated cycles of oxidation and reduction. Thus, the rate of elutriation increased ~35-fold between the 1st and 10th cycle. At an impact velocity of 38. m/s, the amount of fragmentation in the impact test, viz. mass fraction of particles after impact having a size less than that before impact, increased from ~2.3. wt% (fresh particles) to 98. wt% after the 10th cycle. The CuO particles, in comparison, were able to withstand repeated reaction: no signs of increased rates of elutriation or fragmentation were observed over ten cycles. These results highlight the importance of selecting a durable support for oxygen-carriers. © 2011 Elsevier Ltd.

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Segregation or de-blending of bulk particulates is a problem that is encountered in many industrial sectors. The magnitude of segregation can often determine whether a complete production batch can be transferred for onward processing within the plant or released to market. It is a phenomenon that impacts directly upon the profitability of a process. Segregation can occur through a coincidence of a range of variables that relate to the process and bulk particulate properties, common mechanisms for this include; percolation, surface effect (rolling) and elutriation. The importance to industry of predicting the sensitivity of bulk particulates to segregation cannot be under-estimated, and to this end various test procedures have been developed. Within many industries striving to improve product quality and reduce wastage, the determination of variability in blend consistency caused by segregation is an increasing priority. This paper considers recent work undertaken to evaluate the effects of multiple handling operations on the degree of segregation that results. The bulk properties of segregability (and resulting flowability) can not only influence the product consistency, but can have great influence over the process (production) control and performance.

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Ex vivo T cell depletion of allogeneic grafts is associated with a high (up to 80%) rate of mixed chimerism (MC) posttransplantation. The number of transplanted progenitor cells is an important factor in achieving complete donor chimerism in the T cell depletion setting. Use of granulocyte colony-stimulating factor (G-CSF) peripheral blood allografts allows the administration of large numbers of CD34+ cells. We studied the chimeric status of 13 patients who received allogeneic CD34+-selected peripheral blood progenitor cell transplants (allo-PBPCTs/CD34+) from HLA-identical sibling donors. Patients were conditioned with cyclophosphamide (120 mg/kg) and total-body irradiation (13 Gy in four fractions). Apheresis products were T cell-depleted by the immunoadsorption avidin-biotin method. The median number of CD34+ and CD3+ cells infused was 2.8x10(6)/kg (range 1.9-8.6x10(6)/kg) and 0.4x10(6)/kg (range 0.3-1x10(6)/kg), respectively. Molecular analysis of the engraftment was performed using polymerase chain reaction (PCR) amplification of highly polymorphic short tandem repeat (PCR-STR) sequences in peripheral blood samples. MC was detected in two (15%) of 13 patients. These two patients relapsed at 8 and 10 months after transplant, respectively. The remaining 11 patients showed complete donor chimerism and were in clinical remission after a maximum follow-up period of 24 months (range 6-24 months). These results were compared with those obtained in 10 patients who were treated with T cell-depleted bone marrow transplantation by means of elutriation and who received the same conditioning treatment and similar amounts of CD3+ cells (median 0.45x10(6)/kg; not significant) but a lower number of CD34+ cells (median 0.8x10(6)/kg; p = 0.001). MC was documented in six of 10 patients (60%), which was significantly higher than in the allo-PBPCT/CD34+ group (p = 0.04). We conclude that a high frequency of complete donor chimerism is achieved in patients receiving allo-PBPCT/CD34+ and that this is most likely due to the high number of progenitor cells administered.

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Desde tempos históricos que diferentes tipos de lama são utilizados para aplicações externas no corpo humano, para fins terapêuticos e cosméticos. As lamas cuja beneficiação e caracterização físico-química são apresentadas nesta dissertação são formadas no ambiente hipersalino que existe nas salinas da Troncalhada e de São Tiago da Fonte, localizadas no estuário do rio Vouga, em Aveiro, Portugal. As salinas são constituídas por uma sequência de tanques onde, no período de verão, da água do mar e por evaporação natural, se produz sal marinho que precipita no último tanque (o cristalizador) do qual é extraído. Na base dos tanques ocorrem lamas que correspondem a sedimentos constituídos por material argiloso de cor preto-cinzento e que incorporam argila, silte, areia, bioclastos, sal, matéria orgânica e gás. A componente inorgânica da lama extraída do cristalizador foi estudada por Difracção de Raios-X (DRX) e Fluorescência de Raios-X (FRX), enquanto a componente orgânica da mesma lama foi estudada por Cromatografia de Gás- Espectrometria de Massa (GC-MS). Os estudos efectuados incidiram sobre amostras representativas de lama, obtidas antes e após refinação e beneficiação a que foi submetida a lama tal-qual colhida nas salinas. Foram utilizados métodos geofísicos para caracterizar e distinguir as lamas depositadas na época de safra e no período de interregno. Para o efeito, foram cravados tubos amostradores no sedimento que reveste o fundo dos tanques, tubos que seguidamente foram transportados para o laboratório para medição da condutividade eléctrica do topo até à base da coluna de sedimento amostrado. A refinação foi efectuada por elutriação de suspensões aquosas de lama utilizando um equipamento desenvolvido para o efeito e que permitiu concentrar a lama fina no overflow. Após floculação, sifonagem da água sobrenadante, dessalinização e centrifugação a lama refinada e beneficiada pôde ser incorporada em formulações com objectivos terapêuticos e cosméticos. O estudo microbiológico efectuado nas amostras de lama com sal e sem sal e na água das salinas permitiu identificar diversos tipos de bactérias e colónias presentes na lama e avaliar também os processos de esterilização testados. Concluiu-se que a lama hipersalina ou dessalinizada resultante do processamento a que foram submetidas, não deve ser aplicada ou incorporada em formulações tal-qual, pelo facto de em ambas terem sido identificadas bactérias como é o caso de Clostridium perfringens. Não obstante, se submetidas a esterilização utilizando autoclave a lama salina refinada e beneficiada poderá ser aplicada como peloide extemporâneo. Assim sendo foram desenvolvidas formulações dermoterapêuticas e dermocosméticas contendo lama beneficiada e dessalinizada e esterilizada termicamente.

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In this research, the drying process of acerola waste was investigated by using a spouted bed drier. The process was conducted using high density polyethylene inert particles with the objective of producing an ascorbic acid-rich final product. The fruit waste was ground and used to prepare different water-maltodextrin suspensions. Initially, fluidynamical experiments were conducted in order to evaluate the feeding effect on the spouted bed drier fluidynamics behavior. The experimental planning 23 + 3 was used to investigate the effect of the following variables: solids concentration, drying air temperature, intermittence time, production efficiency, solids retention and product losses by elutriation of fine particles on drier walls. The effect of selected independent variables on the drier stability was also evaluated based on a parameter defined as the ratio between the feed suspension volume and the total inert particles volume. Finally, the powder quality was verified in experiments with fixed feed flow and varying air drying temperature, drying air velocity and intermittence time. It was observed that the suspension interferes in the spouted bed drier fluidynamics behavior, and higher air flow is necessary to stabilize the drier. The suspension also promotes the expansion of the spouted bed diameter, decreases the solid circulation and favors the air distribution at the flush area. All variables interfere in the spouted bed performance, and the solids concentration has a major effect on the material retention and losses. The intermittence time also has great effect on the stability and material retention. When it comes to production efficiency, the main effect observed was the drying air temperature. First order models were well adjusted to retention and losses data. The acerola powder presented ascorbic acid levels around 600 to 700 mg/100g. Similar moisture and ascorbic acid levels were obtained for powders obtained by spouted bed and spray drier. However, the powder production efficiency of the spray drier was lower when compared to spouted bed drier. When it comes to energetic analysis, the spray drier process was superior. The results obtained for spouted bed drier are promising and highly dependent on the operational parameters chosen, but in general, it is inferred that this drying process is adequate for paste and suspension drying

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This work presents experimental information relevant to the combustion of biomass in a bubbling fluidized bed. The biomass distribution in a fluidized bed was studied through tests performed in a cold bed, while the volatiles released in the biomass pyrolysis, the burning rate of the resulting charcoal, and the combustion control regime, were studied through tests performed in a high temperature bed.Visual examination of photographs taken from a transparent walls bed, with a rectangular cross-section, showed that the large fuel particles, typical of biomass processing, were distributed in the bubbles, in the splash zone, and in the emulsion phase. The occurrence of biomass in the emulsion phase was favored by burning biomass particles of greater density and smaller size-expetimentally determined in each case. Decreasing the fuel particle size improved the biomass distribution inside the bed. The same was accomplished by increasing the superficial gas velocity as high as possible, compatibly with the acceptable elutriation.Burning tests showed that the biomass fuels have the advantage of reaching the diffusional regime at temperatures that can be lower than 1000 K, which ensures that the biomass fuels burn in a stable regime. (C) 2007 Elsevier B.V. All rights reserved.

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Pós-graduação em Engenharia e Ciência de Alimentos - IBILCE

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Die Untersuchungen der murinen Cytomegalovirus (mCMV) Infektion im BALB/c Mausmodell konzentrierten sich bislang auf die Lunge, da diese einen Hauptort der mCMV Latenz darstellt. Da latentes CMV auch häufig durch Lebertransplantationen übertragen wird, wurde in dieser Arbeit die Leber als ein weiteres medizinisch relevantes Organ der CMV Latenz und Reaktivierung untersucht. Um zunächst die zellulären Orte der mCMV Latenz in der Leber zu ermitteln, wurden verschiedengeschlechtliche Knochenmarktransplantationen (KMT) mit männlichen tdy-positiven Spendern und weiblichen, tdy-negativen Empfängern, mit anschließender mCMV Infektion durchgeführt, um latent infizierte Mäuse mit geschlechtschromosomalem Chimärismus zu generieren. Diese Chimären erlaubten eine Unterscheidung zwischen tdy-positiven Zellen hämatopoetischen Ursprungs und tdy-negativen stromalen und parenchymalen Gewebszellen. Die Separation von Leberzellen der Chimären mittels zentrifugaler Elutriation und anschließender DNA Quantifizierung viraler und zellulärer Genome durch eine quantitative real-time PCR ergab einen ersten Hinweis, dass Endothelzellen ein zellulärer Ort der mCMV Latenz sind. Die darauf folgende immunomagnetische Zelltrennung lokalisierte latente virale DNA in der CD31-positiven Zellfraktion. Die Koexpression von CD31 mit dem endothelzellspezifischen Oberflächenmarker ME-9F1 identifizierte die sinusoidalen Endothelzellen der Leber (LSEC) als die Zellen, die latente virale DNA beherbergen. In den zytofluorometrisch aufgereinigten CD31+/ME-9F1+ LSEC waren bei gleichzeitigem Rückgang der männlichen tdy Markergene virale Genome angereichert, was darauf hinwies, dass Zellen, die virale DNA enthalten, vom Knochenmark-Empfänger stammen. Durch zytofluorometrische Analysen isolierter LSEC konnte eine vom Spender abstammende Subpopulation MHCII+/CD11b+ LSEC identifiziert werden. Anschließende Quantifizierungen viraler DNA aus latent infizierten Mäusen detektierten eine Abnahme viraler Genome mit zunehmender Menge an tdy-positiven Zellen, was beweist, dass MHCII+/CD11b+ LSEC keinen Ort der mCMV Latenz darstellen. Die limiting dilution Untersuchungen der isolierten latent infizierten LSEC ergaben eine Frequenz von einer latent infizierten Zelle unter ~1,9x104 LSEC und eine Anzahl von 7 bis 19 viralen Genomen pro latent infizierter Zelle. Nach 24 Stunden Kultivierung der LSEC konnte mittels quantitativer real-time RT-PCR mit Gesamt-RNA aus LSEC ein Anstieg der Genexpression der immediate early Gene ie1 und ie3 sowie eine Induktion des early Gens e1 gezeigt werden. Eine Erhöhung der transkriptionellen Reaktivierung durch die Inkubation der LSEC mit unterschiedlichen HDAC Inhibitoren konnte allerdings nicht erzielt werden, da sowohl die Menge der isolierten RNA aus behandelten Kulturen, als auch die Anzahl viraler Transkripte im Vergleich zu den unbehandelten Kulturen erniedrigt war. Aufgrund der kurzen Lebensdauer isolierter LSEC in vitro konnte durch Kokultivierungen latent infizierter LSEC zusammen mit murinen embryonalen Fibroblasten keine Virusreaktivierung induziert werden. Im Gegensatz dazu wurden durch den Transfer gereinigter ME-9F1+/CD31+ LSEC aus latent infizierten Spendern in immunsupprimierte Empfänger virale Rekurrenzen in Lungenexplantatkulturen des Rezipienten detektiert. Damit konnten LSEC eindeutig als zellulärer Ort von mCMV Latenz und Reaktivierung in der Leber identifiziert werden.

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Volcanic ash clouds can be fed by an upward-directed eruption column (Plinian column) or by elutriation from extensive pyroclastic-flows (coignimbrite cloud). For large-scale eruptions, there is considerable uncertainty about which mechanism is dominant. Here we analyze in a novel way a comprehensive grainsize database for pyroclastic deposits. We demonstrate that the Mount Pinatubo climactic eruption deposits were substantially derived from coignimbrite clouds, and not only by a Plinian cloud as generally thought. Coignimbrite ash-fall deposits are much richer in breathable <10 m ash (5–25 wt%) than pure Plinian ash at most distances from the source volcano. We also show that coignimbrite ash clouds, as at Pinatubo, are expected to be more water rich than Plinian clouds, leading to removal of more HCl prior to stratospheric injection, thereby reducing their atmospheric impact.