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191.
Chiarella P Leuener M Fasci C de Marco A Santini MP Fazio VM Sawyer AM 《Biotechnology progress》2011,27(2):571-576
We have previously demonstrated how to transform the conventional method of hybridoma production and screening into a fast, high-throughput technology. Nevertheless, there were still open questions related to automated procedures and immunization protocols that we address now by comparing the hybridoma production work-flow in automated and manually executed processes. In addition, since the animals' antibody responses to single or multiple antigen challenge affect monoclonal antibody throughput, different immunization and fusion strategies were tested. Specifically, the results obtained with multiplexing (multiple target antigens injected into a single animal) and single antigen immunization followed by splenocyte pooling immediately before fusion were compared with conventional methods. The results presented here demonstrate that the optimal protocol consists of automated somatic-cell fusion and hybridoma dilution followed by manual plating of hybridoma cells. Additionally, more specific and productive hybridoma clones were obtained with multiplexed immunization in a single animal with respect to the splenocyte pooling from single antigen immunized animals. However, in terms of overall antibody yield, the conventional method consisting of single immunization for each single animal assured ten times more specific hybridoma cell lines than the strategy based on the multiple antigen immunization followed by separate fusion step. In conclusion, the most productive approach for recovering a large number of suitable antibodies relies on single antigen immunization followed by automated fusion and dilution steps and manual plating. 相似文献
192.
Sebastio Perrini Romina Ficarella Ernesto Picardi Angelo Cignarelli Maria Barbaro Pasquale Nigro Alessandro Peschechera Orazio Palumbo Massimo Carella Michele De Fazio Annalisa Natalicchio Luigi Laviola Graziano Pesole Francesco Giorgino 《PloS one》2013,8(3)
Differences in the inherent properties of adipose tissue-derived stem cells (ASC) may contribute to the biological specificity of the subcutaneous (Sc) and visceral (V) adipose tissue depots. In this study, three distinct subpopulations of ASC, i.e. ASCSVF, ASCBottom, and ASCCeiling, were isolated from Sc and V fat biopsies of non-obese subjects, and their gene expression and functional characteristics were investigated. Genome-wide mRNA expression profiles of ASCSVF, ASCBottom and ASCCeiling from Sc fat were significantly different as compared to their homologous subsets of V-ASCs. Furthermore, ASCSVF, ASCCeiling and ASCBottom from the same fat depot were also distinct from each other. In this respect, both principal component analysis and hierarchical clusters analysis showed that ASCCeiling and ASCSVF shared a similar pattern of closely related genes, which was highly different when compared to that of ASCBottom. However, larger variations in gene expression were found in inter-depot than in intra-depot comparisons. The analysis of connectivity of genes differently expressed in each ASC subset demonstrated that, although there was some overlap, there was also a clear distinction between each Sc-ASC and their corresponding V-ASC subsets, and among ASCSVF, ASCBottom, and ASCCeiling of Sc or V fat depots in regard to networks associated with regulation of cell cycle, cell organization and development, inflammation and metabolic responses. Finally, the release of several cytokines and growth factors in the ASC cultured medium also showed both inter- and intra-depot differences. Thus, ASCCeiling and ASCBottom can be identified as two genetically and functionally heterogeneous ASC populations in addition to the ASCSVF, with ASCBottom showing the highest degree of unmatched gene expression. On the other hand, inter-depot seem to prevail over intra-depot differences in the ASC gene expression assets and network functions, contributing to the high degree of specificity of Sc and V adipose tissue in humans. 相似文献
193.
Babaev VR Whitesell RR Li L Linton MF Fazio S May JM 《Free radical biology & medicine》2011,50(1):27-36
To test whether severe ascorbic acid deficiency in macrophages affects progression of early atherosclerosis, we used fetal liver cell transplantation to generate atherosclerosis-prone apolipoprotein E-deficient (apoE(-/-)) mice that selectively lacked the ascorbate transporter (SVCT2) in hematopoietic cells, including macrophages. After 13 weeks of chow diet, apoE(-/-) mice lacking the SVCT2 in macrophages had surprisingly less aortic atherosclerosis, decreased lesion macrophage numbers, and increased macrophage apoptosis compared to control-transplanted mice. Serum lipid levels were similar in both groups. Peritoneal macrophages lacking the SVCT2 had undetectable ascorbate; increased susceptibility to H(2)O(2)-induced mitochondrial dysfunction and apoptosis; decreased expression of genes for COX-2, IL1β, and IL6; and decreased lipopolysaccharide-stimulated NF-κB and antiapoptotic gene expression. These changes were associated with decreased expression of both the receptor for advanced glycation end products and HIF-1α, either or both of which could have been the proximal cause of decreased macrophage activation and apoptosis in ascorbate-deficient macrophages. 相似文献
194.
195.
V. Carcangiu C. Giannetto S. Luridiana F. Fazio M. C. Mura A. Parmeggiani 《Biological Rhythm Research》2017,48(1):35-41
The aim of the study was to investigate the influence of early lactation on leptin and growth hormone (GH) during bathyphase. Forty lactating Sarda ewes were divided into two equal groups on the basis of their milk production levels: HIGH (>1100 g/day) and LOW (<900 g/day). From the 5th to the 110th day after lambing, every 15 days, body condition score (BCS) was recorded and milk samples were collected. At the same data point, blood sampling was performed and leptin, GH and, Non-Esterified Fatty Acids (NEFA) were assessed. On milk, fat and proteins were determined. Statistical differences were observed in BCS, leptin, GH, NEFA and fat concentration in milk between the two groups. BCS was lower in high group and decreased from the 20th to the 90th day of lactation. Leptin was higher in low group and increased from the 50th and the 65th day of lactation, in low and high groups, respectively. GH and NEFA were higher in high group and decreased from the mild lactation. In high group, BCS and milk yield were negatively correlated and leptin was negatively correlated with GH and NEFA. In low group, leptin was positively correlated with BCS and negatively correlated with the all studied parameters. GH and NEFA were positively correlated with both groups. In conclusion, milk yield had an effect on the leptin and GH plasma values recorded during their bathyphase. 相似文献
196.
Rute ST Martins Laurence AM Deloffre Constantinos C Mylonas Deborah M Power Adelino VM Canário 《Reproductive biology and endocrinology : RB&E》2007,5(1):19
Background
DAX1 (NR0B1), a member of the nuclear receptors super family, has been shown to be involved in the genetic sex determination and in gonadal differentiation in several vertebrate species. In the aquaculture fish European sea bass, Dicentrarchus labrax, and in the generality of fish species, the mechanisms of sex determination and differentiation have not been elucidated. The present study aimed at characterizing the European DAX1 gene and its developmental expression at the mRNA level. 相似文献197.
Raffaela Barbano Orazio Palumbo Barbara Pasculli Marco Galasso Stefano Volinia Vincenzo D'Angelo Nadia Icolaro Michelina Coco Lucia Dimitri Paolo Graziano Massimiliano Copetti Vanna Maria Valori Evaristo Maiello Massimo Carella Vito Michele Fazio Paola Parrella 《PloS one》2014,9(10)
Gliomas represent a disparate group of tumours for which there are to date no cure. Thus, there is a recognized need for new diagnostic and therapeutic approaches based on increased understanding of their molecular nature. We performed the comparison of the microRNA (miRNA) profile of 8 WHO grade II gliomas and 24 higher grade tumours (2 WHO grade III and 22 glioblastomas) by using the Affymetrix GeneChip miRNA Array v. 1.0. A relative quantification method (RT-qPCR) with standard curve was used to confirm the 22 miRNA signature resulted by array analysis. The prognostic performances of the confirmed miRNAs were estimated on the Tumor Cancer Genome Atlas (TCGA) datasets. We identified 22 miRNAs distinguishing grade II gliomas from higher grade tumours. RT-qPCR confirmed the differential expression in the two patients'' groups for 13 out of the 22 miRNAs. The analysis of the Glioblastoma Multiforme (GBM) and Lower Grade Glioma (LGG) datasets from TCGA demonstrated the association with prognosis for 6 of those miRNAs. Moreover, in the GBM dataset miR-21 and miR-210 were predictors of worse prognosis in both univariable and multivariable Cox regression analyses (HR 1.19, p = 0.04, and HR 1.18, p = 0.029 respectively). Our results support a direct contribution of miRNAs to glioma cancerogenesis and suggest that miR-21 and miR-210 may play a role in the aggressive clinical behaviour of glioblastomas. 相似文献
198.
Marjolein?Helder David?PBTB?StrikEmail author Hubertus?VM?Hamelers Cees?JN?Buisman 《Biotechnology for biofuels》2012,5(1):70
Due to a growing world population and increasing welfare, energy demand worldwide is increasing. To meet the increasing energy demand in a sustainable way, new technologies are needed. The Plant-Microbial Fuel Cell (P-MFC) is a technology that could produce sustainable bio-electricity and help meeting the increasing energy demand. Power output of the P-MFC, however, needs to be increased to make it attractive as a renewable and sustainable energy source. To increase power output of the P-MFC internal resistances need to be reduced. With a flat-plate P-MFC design we tried to minimize internal resistances compared to the previously used tubular P-MFC design. With the flat-plate design current and power density per geometric planting area were increased (from 0.15 A/m2 to 1.6 A/m2 and from 0.22 W/m2 to and 0.44 W/m2)as were current and power output per volume (from 7.5 A/m3 to 122 A/m3 and from 1.3 W/m3 to 5.8 W/m3). Internal resistances times volume were decreased, even though internal resistances times membrane surface area were not. Since the membrane in the flat-plate design is placed vertically, membrane surface area per geometric planting area is increased, which allows for lower internal resistances times volume while not decreasing internal resistances times membrane surface area. Anode was split into three different sections on different depths of the system, allowing to calculate internal resistances on different depths. Most electricity was produced where internal resistances were lowest and where most roots were present; in the top section of the system. By measuring electricity production on different depths in the system, electricity production could be linked to root growth. This link offers opportunities for material-reduction in new designs. Concurrent reduction in material use and increase in power output brings the P-MFC a step closer to usable energy density and economic feasibility. 相似文献
199.
Maya?P?Piddocke Alessandro?Fazio Wanwipa?Vongsangnak Man?L?Wong Hans?P?Heldt-Hansen Chris?Workman Jens?Nielsen Lisbeth?OlssonEmail author 《Microbial cell factories》2011,10(1):27
Background
Addition of sugar syrups to the basic wort is a popular technique to achieve higher gravity in beer fermentations, but it results in dilution of the free amino nitrogen (FAN) content in the medium. The multicomponent protease enzyme Flavourzyme has beneficial effect on the brewer's yeast fermentation performance during high gravity fermentations as it increases the initial FAN value and results in higher FAN uptake, higher specific growth rate, higher ethanol yield and improved flavour profile. 相似文献200.
Redouan Bshary Rui F Oliveira Tânia SF Oliveira Adelino VM Canário 《Frontiers in zoology》2007,4(1):1-8