scholarly journals Transcriptomic analyses of cacao flavonoids produced in photobioreactors

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Adriana M. Gallego ◽  
Luisa F. Rojas ◽  
Wilmar G. Valencia ◽  
Lucía Atehortúa ◽  
Aura I. Urrea ◽  
...  

Abstract Background Theobroma cacao is a major source of flavonoids such as catechins and their monomers proanthocyanidins (PAs), widely studied for their potential benefits in cardiovascular diseases. Light has been shown to promote plant secondary metabolite production in vitro. In this study, cacao cells cultured in 7.5 L stirred tank photobioreactors (STPs) were exposed to a change of white to blue LED lights for 28 days (d). Results Transcriptomic analyses were performed in three time points comparing changing expression patterns, after cell exposure to white light (d0-VS-d14), after a shift from white to blue light (d14-VS-d15), and after an extended period of blue light for the following 15 days (d15-VS-d28). Under white light, there was enrichment in metabolic pathways associated with cell growth (carbon, glycolysis, and amino acid biosynthesis) accompanied by a significant increase in the PAs content. In the shift to blue light, further increase in PAs content was observed concomitantly with the significant expression of TWO-COMPONENT RESPONSE REGULATOR genes involved in the early stress responses via circadian clock and hormone pathways. Under blue light exposure, we observed a depletion of PAs content associated with ROS-mediated stress pathways. Conclusions Light effects on large-scale cell cultures in photobioreactors are complex and pleiotropic; however, we have been able to identify key regulatory players upstream cacao flavonoid biosynthesis in STPs, including TWO-COMPONENT SYSTEM and ROS-signaling genes. The crosstalk between flavonoid biosynthesis and regulatory networks led to understand the dynamics of flavonoid production and degradation in response to light-driven ROS signals. This can be used to optimize the time, and the yield of in vitro targeted metabolites in large-scale culture systems.

2015 ◽  
Vol 14 (6) ◽  
pp. 1093-1099 ◽  
Author(s):  
Pei Zhang ◽  
Chen Huang ◽  
Wei Wang ◽  
Minshu Wang

In vitro nonlethal blue light exposure led to increases in oxidative stress and mitochondrial content staurosporine-differentiated RGC-5 cells, which as the early response to lower dose of blue light exposure.


Blood ◽  
1998 ◽  
Vol 92 (12) ◽  
pp. 4622-4631 ◽  
Author(s):  
William L. Stanford ◽  
Georgina Caruana ◽  
Katherine A. Vallis ◽  
Maneesha Inamdar ◽  
Michihiro Hidaka ◽  
...  

Abstract We have developed a large-scale, expression-based gene trap strategy to perform genome-wide functional analysis of the murine hematopoietic and vascular systems. Using two different gene trap vectors, we have isolated embryonic stem (ES) cell clones containing lacZreporter gene insertions in genes expressed in blood island and vascular cells, muscle, stromal cells, and unknown cell types. Of 79 clones demonstrating specific expression patterns, 49% and 16% were preferentially expressed in blood islands and/or the vasculature, respectively. The majority of ES clones that expressedlacZ in blood islands also expressed lacZ upon differentiation into hematopoietic cells on OP9 stromal layers. Importantly, the in vivo expression of the lacZ fusion products accurately recapitulated the observed in vitro expression patterns. Expression and sequence analysis of representative clones suggest that this approach will be useful for identifying and mutating novel genes expressed in the developing hematopoietic and vascular systems.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kemal Avican ◽  
Jehad Aldahdooh ◽  
Matteo Togninalli ◽  
A. K. M. Firoj Mahmud ◽  
Jing Tang ◽  
...  

AbstractBacterial processes necessary for adaption to stressful host environments are potential targets for new antimicrobials. Here, we report large-scale transcriptomic analyses of 32 human bacterial pathogens grown under 11 stress conditions mimicking human host environments. The potential relevance of the in vitro stress conditions and responses is supported by comparisons with available in vivo transcriptomes of clinically important pathogens. Calculation of a probability score enables comparative cross-microbial analyses of the stress responses, revealing common and unique regulatory responses to different stresses, as well as overlapping processes participating in different stress responses. We identify conserved and species-specific ‘universal stress responders’, that is, genes showing altered expression in multiple stress conditions. Non-coding RNAs are involved in a substantial proportion of the responses. The data are collected in a freely available, interactive online resource (PATHOgenex).


Blood ◽  
1998 ◽  
Vol 92 (12) ◽  
pp. 4622-4631 ◽  
Author(s):  
William L. Stanford ◽  
Georgina Caruana ◽  
Katherine A. Vallis ◽  
Maneesha Inamdar ◽  
Michihiro Hidaka ◽  
...  

We have developed a large-scale, expression-based gene trap strategy to perform genome-wide functional analysis of the murine hematopoietic and vascular systems. Using two different gene trap vectors, we have isolated embryonic stem (ES) cell clones containing lacZreporter gene insertions in genes expressed in blood island and vascular cells, muscle, stromal cells, and unknown cell types. Of 79 clones demonstrating specific expression patterns, 49% and 16% were preferentially expressed in blood islands and/or the vasculature, respectively. The majority of ES clones that expressedlacZ in blood islands also expressed lacZ upon differentiation into hematopoietic cells on OP9 stromal layers. Importantly, the in vivo expression of the lacZ fusion products accurately recapitulated the observed in vitro expression patterns. Expression and sequence analysis of representative clones suggest that this approach will be useful for identifying and mutating novel genes expressed in the developing hematopoietic and vascular systems.


1981 ◽  
Vol 15 ◽  
pp. 669-669
Author(s):  
L D Lilien ◽  
S Voora ◽  
G Srinivasan ◽  
R S Pildes
Keyword(s):  

2021 ◽  
Author(s):  
Maomei Luo ◽  
Chun Zeng ◽  
Shu Wang ◽  
Shanjun Cai

Abstract AimsTo establish the N-retinylidene-N-retinylethanolamine(A2E) and blue light induced RPE cells damage model to explore the regularity of distribution of A2E and the levels of reactive oxygen species(ROS).MethodsThe fourth to sixth generation of human RPE cells in vitro were divided into five groups randomly: control group, blue light group, A2E-loaded group, A2E-loaed+blue light group and A2E-loaded+blue light +nifedipine group. The levels of ROS in cytoplasm by DCFH-DA staining was assayed by flow cytometry. The concentration of A2E in cytoplasm and lysosomes were assayed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The fluorescence intensity of A2E in lysosomes by Lysotracker redDND-99 staining was assayed by confocal laser scanning microscope. ResultsExposure to blue light and/or A2E could increase the levels of ROS in RPE cells, and nifedipine could inhibit oxidative stress response and reduce ROS levels. By HPLC-MS, it was found that A2E was not detected in the groups without load A2E, and A2E levels in cytoplasm and lysosomes decreased after light exposure. The green fluorescence produced by A2E loaded on RPE cells was mostly coincident with the red fluorescence labeled by lysosomes.ConclusionBlue light and A2E can increase the ROS levels of RPE cells and both have a synergistic effect. A2E is mainly concentrated in lysosomes, which is reduced by oxidation under blue light irradiation, damages lysosomal membrane with oxidized species of A2E, and leaks out from lysosomes.


2019 ◽  
Author(s):  
M. Ashley Sobran ◽  
Peggy A. Cotter

AbstractTo detect and respond to the diverse environments they encounter, bacteria often use two-component regulatory systems (TCSs) to coordinate essential cellular processes required for survival. In pathogenicBordetellaspecies, the BvgAS TCS regulates expression of hundreds of genes, including those encoding all known protein virulence factors, and its kinase activity is essential for respiratory infection. Maintenance of BvgS kinase activity in the lower respiratory tract (LRT) depends on the function of another TCS, PlrSR. While the periplasmic venus fly-trap domains of BvgS have been implicated in responding to so-called modulating signalsin vitro(nicotinic acid and MgSO4), a role for the cytoplasmic Per-Arnt-Sim (PAS) domain in signal perception has not previously been demonstrated. By comparingB. bronchisepticastrains with mutations in the PAS domain-encoding region ofbvgSwith wild-type bacteriain vitroandin vivo, we found that although the PAS domain is not required to sense modulating signalsin vitro, it is required for the inactivation of BvgS that occurs in the absence of PlrS in the LRT of mice, suggesting that the BvgS PAS domain functions as an independent signal perception domain. Our data also indicate that the BvgS PAS domain is important for controlling absolute levels of BvgS kinase activity and the efficiency of the response to modulating signalsin vitro. Our results indicate that BvgS is capable of integrating sensory inputs from both the periplasm and the cytoplasm to control precise gene expression patterns in diverse environmental conditions.ImportanceDespite high rates of vaccination, Pertussis, a severe, highly contagious respiratory disease, caused by the bacteriumBordetella pertussis, has reemerged as a significant health threat. InBordetella pertussisand the closely related species,Bordetella bronchiseptica, activity of the BvgAS two-component regulatory system is critical for colonization of the human respiratory tract and other mammalian hosts, respectively. Here we show that the cytoplasmic PAS domain of BvgS can function as an independent signal perception domain that is capable of integrating environmental signals that influence overall BvgS activity. Our work is significant as it reveals a critical, yet previously unrecognized role, for the PAS domain in the BvgAS phosphorelay and provides a greater understanding of virulence regulation inBordetella.


HortScience ◽  
2015 ◽  
Vol 50 (3) ◽  
pp. 430-433 ◽  
Author(s):  
Fang Geng ◽  
Renae Moran ◽  
Michael Day ◽  
William Halteman ◽  
Donglin Zhang

The influence of red and blue light wavelengths was tested to improve the initial in vitro multiplication of apple (Malus × domestica) rootstock cultivars Budagovsky 9 (B.9), Geneva 30 (G.30), and Geneva 41 (G.41). Single-node segments were established in semisolid Murashige and Skoog media and then transferred to proliferation media and cultured 40 days under white, red, or blue light irradiance. In a second experiment, G.30 was cultured under red, blue, or white light with and without gibberellic acid (GA3). The three rootstocks responded similarly under white light in terms of shoot number, length of the longest shoot, and the number of elongated shoots. Red light increased the number of shoots, length of the longest shoot, and the number of elongated shoots of B.9 and G.30 when compared with white or blue light. Red light increased the number of elongated B.9 and G.30 shoots to five per explant compared with one per explant under white light. In contrast, shoot growth of G.41 showed no difference under the three light quality treatments, and the number of elongated shoots per explant was less than one. When compared with an absence of GA3, a concentration of GA3 at 0.5 mg·L−1 promoted in vitro shoot growth of G.30 under red and blue light.


2019 ◽  
Author(s):  
Corey G. Duke ◽  
Katherine E. Savell ◽  
Robert A. Phillips ◽  
Jeremy J. Day

Blue waveform light is used as an optical actuator in numerous optogenetic technologies employed in neuronal systems. However, the potential side effects of blue waveform light in neurons has not been thoroughly explored, and recent reports suggest that neuronal exposure to blue light can induce transcriptional alterations in vitro and in vivo. Here, we examined the effects of blue waveform light in cultured primary rat cortical neurons. Exposure to blue light (470nm) resulted in upregulation of several immediate early genes (IEGs) traditionally used as markers of neuronal activity, including Fos and Fosb, but did not alter the expression of circadian clock genes Bmal1, Cry1, Cry2, Clock, or Per2. IEG expression was increased following 4 hours of 5% duty cycle light exposure, and IEG induction was not dependent on light pulse width. Elevated levels of blue light exposure induced a loss of cell viability in vitro, suggestive of overt phototoxicity. Changes in gene expression induced by blue waveform light were prevented when neurons were cultured in a photoinert media supplemented with a photostable neuronal supplement instead of commonly utilized neuronal culture media and supplements. Together, these findings suggest that light-induced gene expression alterations observed in vitro stem from a phototoxic interaction between commonly used media and neurons, and offer a solution to prevent this toxicity when using photoactivatable technology in vitro.


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