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Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes
Mark F Miller, Rita Loch-Caruso
Reproductive Biology and Endocrinology , 2010, DOI: 10.1186/1477-7827-8-121
Abstract: Tissue samples were exposed to 100 ng/ml LPS for 12 h and cytokines were measured by ELISA. Data are expressed as increase relative to non-treated controls.Levels of interleukin-6 increased in punch culture medium samples to a significantly greater extent (34.2 fold) compared with medium from transwell cultures in the amnion (6.6 fold) or choriodecidual (7.1 fold) compartments. Interleukin-8 also showed a significantly greater induction in punch (4.8 fold) than transwell amnion (1.6 fold) or choriodecidual (1.7 fold) samples. The anti-inflammatory interleukin-10 showed a significant difference between punch (36.5 fold) and transwell amnion (15.4 fold) samples, but no difference was observed between punch and transwell choriodecidual (28.5 fold) samples. Neither interleukin-1beta nor tumor necrosis factor-alpha (TNF-alpha) showed a significant difference between the punch and transwell samples.These results indicate that the pattern of LPS-stimulated cytokine release from gestational membranes in vitro depends on the culture system used, confounding comparisons of studies that use different gestational membrane culture systems to study inflammatory responses.Gestational membranes (amnion, chorion laeve and decidua) collected immediately after birth and cultured in vitro allow assessment of responses in tissues with an intact cellular matrix. As such, cultures of human gestational membranes provide useful in vitro research models for inquiries into obstetric challenges such as inflammation, preterm premature rupture of membranes (PPROM) and preterm birth.One model system used extensively to study stimulated production and release of cytokines in human gestational membranes in vitro involves explant culture of a biopsy punch, with the gestational tissue punch explant free-floating in culture medium. Biopsy punch explant cultures may use full-thickness membranes [1,2] as well as separated amnion [3,4] or choriodecidua [5,6]. This single-compartment explant culture syste
Offspring from Mouse Embryos Developed Using a Simple Incubator-Free Culture System with a Deoxidizing Agent  [PDF]
Fumiaki Itoi, Mikiko Tokoro, Yukari Terashita, Kazuo Yamagata, Noritaka Fukunaga, Yoshimasa Asada, Teruhiko Wakayama
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0047512
Abstract: To culture preimplantation embryos in vitro, water-jacketed CO2 incubators are used widely for maintaining an optimal culture environment in terms of gas phase, temperature and humidity. We investigated the possibility of mouse embryo culture in a plastic bag kept at 37°C. Zygotes derived from in vitro fertilization or collected from naturally mated B6D2F1 female mice were put in a drop of medium on a plastic culture dish and then placed in a commercially available plastic bag. When these were placed in an oven under air at 37°C for 96 h, the rate of blastocyst development and the cell numbers of embryos decreased. However, when the concentration of O2 was reduced to 5% using a deoxidizing agent and a small oxygen meter, most zygotes developed into blastocysts. These blastocysts were judged normal according to their cell number, Oct3/4 and Cdx2 gene expression levels, the apoptosis rate and the potential for full-term development after embryo transfer to pseudopregnant recipients. Furthermore, using this system, normal offspring were obtained simply by keeping the bag on a warming plate. This culture method was applied successfully to both hybrid and inbred strains. In addition, because the developing embryos could be observed through the transparent wall of the bag, it was possible to capture time-lapse images of live embryos until the blastocyst stage without needing an expensive microscope-based incubation chamber. These results suggest that mouse zygotes are more resilient to their environment than generally believed. This method might prove useful in economical culture systems or for the international shipment of embryos.
Simple system using natural mineral water for high-throughput phenotyping of Arabidopsis thaliana seedlings in liquid culture
Benamar A, Pierart A, Baecker V, Avelange-Macherel MH, Rolland A, Gaudichon S, di Gioia L, Macherel D
International Journal of High Throughput Screening , 2013, DOI: http://dx.doi.org/10.2147/IJHTS.S40565
Abstract: ystem using natural mineral water for high-throughput phenotyping of Arabidopsis thaliana seedlings in liquid culture Methodology (236) Total Article Views Authors: Benamar A, Pierart A, Baecker V, Avelange-Macherel MH, Rolland A, Gaudichon S, di Gioia L, Macherel D Published Date February 2013 Volume 2013:4 Pages 1 - 15 DOI: http://dx.doi.org/10.2147/IJHTS.S40565 Received: 21 November 2012 Accepted: 12 January 2013 Published: 04 March 2013 Abdelilah Benamar,1 Antoine Pierart,1 Volker Baecker,2 Marie-Hélène Avelange-Macherel,3 Aurélia Rolland,1 Sabine Gaudichon,4 Lodovico di Gioia,4 David Macherel1 1Université d’Angers, Lunam Université, Angers, 2MRI-Montpellier RIO Imaging, Montpellier, 3Agrocampus Ouest, Angers, 4Danone Research, Palaiseau Cedex, France Background: Phenotyping for plant stress tolerance is an essential component of many research projects. Because screening of high numbers of plants and multiple conditions remains technically challenging and costly, there is a need for simple methods to carry out large-scale phenotyping in the laboratory. Methods: We developed a method for phenotyping the germination and seedling growth of Arabidopsis (Arabidopsis thaliana) Col-0 in liquid culture. Culture was performed under rotary shaking in multiwell plates, using Evian natural mineral water as a medium. Nondestructive and accurate quantification of green pixels by digital image analysis allowed monitoring of growth. Results: The composition of the water prevented excessive root elongation growth that would otherwise lead to clumping of seedlings observed when classic nutrient-rich medium or deionized water is used. There was no need to maintain the cultures under aseptic conditions, and seedlings, which are photosynthetic, remained healthy for several weeks. Several proof-of-concept experiments demonstrated the usefulness of the approach for environmental stress phenotyping. Conclusion: The system described here is easy to set up, cost-effective, and enables a single researcher to screen large numbers of lines under various conditions. The simplicity of the method clearly makes it amenable to high-throughput phenotyping using robotics.
Simple system using natural mineral water for high-throughput phenotyping of Arabidopsis thaliana seedlings in liquid culture  [cached]
Benamar A,Pierart A,Baecker V,Avelange-Macherel MH
International Journal of High Throughput Screening , 2013,
Abstract: Abdelilah Benamar,1 Antoine Pierart,1 Volker Baecker,2 Marie-Hélène Avelange-Macherel,3 Aurélia Rolland,1 Sabine Gaudichon,4 Lodovico di Gioia,4 David Macherel11Université d’Angers, Lunam Université, Angers, 2MRI-Montpellier RIO Imaging, Montpellier, 3Agrocampus Ouest, Angers, 4Danone Research, Palaiseau Cedex, FranceBackground: Phenotyping for plant stress tolerance is an essential component of many research projects. Because screening of high numbers of plants and multiple conditions remains technically challenging and costly, there is a need for simple methods to carry out large-scale phenotyping in the laboratory.Methods: We developed a method for phenotyping the germination and seedling growth of Arabidopsis (Arabidopsis thaliana) Col-0 in liquid culture. Culture was performed under rotary shaking in multiwell plates, using Evian natural mineral water as a medium. Nondestructive and accurate quantification of green pixels by digital image analysis allowed monitoring of growth.Results: The composition of the water prevented excessive root elongation growth that would otherwise lead to clumping of seedlings observed when classic nutrient-rich medium or deionized water is used. There was no need to maintain the cultures under aseptic conditions, and seedlings, which are photosynthetic, remained healthy for several weeks. Several proof-of-concept experiments demonstrated the usefulness of the approach for environmental stress phenotyping.Conclusion: The system described here is easy to set up, cost-effective, and enables a single researcher to screen large numbers of lines under various conditions. The simplicity of the method clearly makes it amenable to high-throughput phenotyping using robotics.Keywords: phenotyping, phenomics, stress, seed, seedling, image analysis
Modified extensive pond culture of Litopenaeus vannamei for sustainable shrimp culture in the Philippines
Cecilia J. Jaspe,Christopher M. A. Caipang,Bessie J. G. Elle
Advances in Environmental Sciences , 2011,
Abstract: The shrimp culture industry provides huge revenues to most aquaculture producing countries,but it is also beset with problems that hamper its sustainability. In the present study, we described amodified extensive pond culture method for white shrimp, Litopenaeus vannamei in the Philippines duringthe wet and dry months. One hectare earthen ponds were prepared and added with organic and/orinorganic fertilizers to stimulate natural food production. The ponds were stocked with L. vannameipostlarvae (PL) at a density of 4 PL m-2. A zero-water exchange system of pond management was doneduring the first two months of culture followed by a bi-weekly water exchange until harvest. No artificialfeeding was given during the culture period, instead, the ponds were applied with inorganic fertilizerevery month to ensure continuous supply of natural food. During the culture period, the shrimpsappeared healthy and no disease outbreaks were observed. All the physico-chemical parameters of thewater in the pond were within the optimum range required for shrimp farming and the phytoplanktonpopulation was predominantly green microalgae (Chlorophyta). The shrimps were harvested after 3 to3.5 months of culture or when they reached an average body weight of 13-15 g, with moderate to highsurvival rates depending on the prevailing climatic conditions.
BIOLOGICAL CHARACTERISTICS OF POLYMYXA BETAE ON SAND CULTURE SYSTEM
砂培体系中甜菜多粘菌生物学特性的研究

Peng Rihe,Han Chenggui,Yang Lin,Liu Yi,
彭日荷
,韩成贵,杨莉莉,刘仪

菌物学报 , 1997,
Abstract: The life cycle of polyals betae is only 7 days on an improved sand culture system. The influences of pH illumination and inocula on infectivity and multiplication of Polymyxa betae are tested. The infection period of secondary zoospores, survival longevity of zoospores and resting spores are also determined under this modified sand culture system.
Evaluation of a modified culture medium for Borrelia burgdorferi sensu lato
Rodríguez, Islay;Lienhard, Reto;Gern, Lise;Veuve, Marie Colette;Jouda, Fatima;Siegrist, Hans H;Fernández, Carmen;Rodríguez, José Enrique;
Memórias do Instituto Oswaldo Cruz , 2007, DOI: 10.1590/S0074-02762007000800017
Abstract: the aim of the present study was to assess the possible use of a modified medium, prepared in the laboratory using the constituents of barbour-stonner-kelly (bsk) medium and medium 199 as base, for the culture of borrelia strains, comparing the growth of individual strains in this medium and in the bsk-h medium, and the protein profile and antigenic characteristics of borrelia proteins expressed in these media. a qualitative evaluation of growth of borrelia species was made with acceptable results (morphology and motility), but during a quantitative evaluation using the three main genospecies of borrelia, the better results were obtained with a b. burgdorferi sensu stricto strain. the modified medium did not enable the growth of a b. afzelii strain. the protein profile and antigenic characteristic of the expressed proteins in the modified medium were studied with satisfactory results. these results suggest the modified medium as an alternative for the cultivation of borrelia strains, with some limitations, in poorly-resourced laboratories.
Indigenous culture as a knowledge system
Hester du Plessis, Gauhar Raza
Tydskrif vir letterkunde , 2004,
Abstract: Indigenous culture as a knowledge system [English] Complex concepts such as cultural identity, gender issues and the effects of colonialism, politics, and power structures on societies form part of the debate around indigenous culture as a knowledge system. This article makes a contribution to the debate by addressing cultural issues encountered during a cross-cultural research project based in India and South Africa. The authors reflected on some of the conceptual issues they grappled with during their research. The project involved the documentation, study and understanding of the extent in which indigenous knowledge systems (IKS) and modern technologies were utilised in the traditional manufacturing processes of artisans in general and potters in particular. The roles and functions of IKS as used during the production of artefacts were included in the study. This perspective was coupled with a study on the artisans' attitude towards and understanding of science (PAUS) while conducting their traditional technological processes. The combined approach provided a method that allowed researchers to develop interventions that capitalised on existing skills, practices and social relationships rather than undermining them, thus contributing to their sustainability. The project, at the same time, focussed on redefining the characteristics of “knowing” (of knowledge) as not just a mere contemplative gaze, but also as a practical activity. By focusing on artisans, the question of knowledge was placed in the two spheres of knowledge production: “theory” (epistemology) and “practice”. This approach attempted to address and discuss some academic notions based on culture; including a variety of aspects that broadly constitute the “concept” of culture. As these notions continuously alter with changing academic insights they are constantly re-defined by academics and researchers. Key Words: Indigenous culture, indigenous knowledge system (IKS), cultural identity, design – traditional Tydskrif vir letterkunde Vol. 41(2) 2004: 85-98
Cultural Adaptation and Reliability Analysis of the Modified Dyspnea Index for the Brazilian Culture
Miura, Cinthya Tamie Passos;Gallani, Maria Cecília Bueno Jayme;Domingues, Gabriela de Barros Leite;Rodrigues, Roberta Cunha Matheus;Stoller, James K.;
Revista Latino-Americana de Enfermagem , 2010, DOI: 10.1590/S0104-11692010000500025
Abstract: this study aims to present the cross-cultural adaptation process of the modified dyspnea index to the brazilian culture and to investigate its content validity and reliability. this process included the steps of translation, back translation and review by two experts to assess semantic, conceptual, idiomatic, cultural and metabolic equivalence. the index of content validity was used to evaluate the extent of inter-observer agreement. a guide to implement the modified dyspnea index was developed and validated. two different professionals assessed the reliability of the brazilian version of the modified dyspnea index, according to the inter-observer equivalence criterion, with 31 patients, indicating a kappa coefficient=0.960 (p<0.001). in conclusion, the brazilian version of mdi presented evidence of interobserver equivalence when applied by different health professionals in the population of cardiac patients.
The Corticostriatal System in Dissociated Cell Culture  [PDF]
Fiona E. Randall,Catherine Vickers,Sarah C. Schock,Gordon W. Arbuthnott
Frontiers in Systems Neuroscience , 2011, DOI: 10.3389/fnsys.2011.00052
Abstract: The sparse connectivity within the striatum in vivo makes the investigation of individual corticostriatal synapses very difficult. Most studies of the corticostriatal input have been done using electrical stimulation under conditions where it is hard to identify the precise origin of the cortical input. We have employed an in vitro dissociated cell culture system that allows the identification of individual corticostriatal pairs and have been developing methods to study individual neuron inputs to striatal neurons. In mixed corticostriatal cultures, neurons had resting activity similar to the system in vivo. Up/down states were obvious and seemed to encompass the entire culture. Mixed cultures of cortical neurons from transgenic mice expressing green fluorescent protein with striatal neurons from wild-type mice of the same developmental stage allowed visual identification of individual candidate corticostriatal pairs. Recordings were performed between 12 and 37 days in vitro (DIV). To investigate synaptic connections we recorded from 69 corticostriatal pairs of which 44 were connected in one direction and 25 reciprocally. Of these connections 41 were corticostriatal (nine inhibitory) and 53 striatocortical (all inhibitory). The observed excitatory responses were of variable amplitude (?10 to ?370 pA, n = 32). We found the connections very secure – with negligible failures on repeated stimulation (approximately 1 Hz) of the cortical neuron. Inhibitory corticostriatal responses were also observed (?13 to ?314 pA, n = 9). Possibly due to the mixed type of culture we found an inhibitory striatocortical response (?14 to ?598 pA, n = 53). We are now recording from neurons in separate compartments to more closely emulate neuroanatomical conditions but still with the possibility of the easier identification of the connectivity.
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