scholarly journals In vitro Gene Expression Assay

2020 ◽  
Author(s):  
2019 ◽  
Vol 88 ◽  
pp. 18-19
Author(s):  
Peter I. Racz ◽  
Inger Brandsma ◽  
Sabine Hartvelt ◽  
Tom Zwetsloot ◽  
Giel Hendriks

Author(s):  
Chukwudi Nze ◽  
Osaretin Albert Taiwo Ebuehi

Aim: This nutrigenomic research study is to investigate the impact of fermented maize (FM) and non-fermented maize (N-FM) diets on the expression of phosphofructokinase-1 (PFK-1) gene in a diabetic state. Methodology: The rats were equally grouped into four for the subsequent two weeks after acclimatization; Group 1 contained streptozotocinized-diabetic rats fed with FM diet (DFM), Group 2 contained streptozotocinized-diabetic rats fed with N-FM diet (DNM), Group 3 contained the normal control rats fed with standard rodent chow (NCG) and Group 4 contained diabetic control rats fed with standard rodent chow (DCG). The total phenol, flavonoid and antioxidant capacity (in vitro) of the maize diets were analyzed. Results: Rats fed the N-FM diet had higher concentration of phenols (73.20±0.9 mg/100 g) and flavonoids (82.83±1.02 mg/100 g). The in vitro antioxidant assay showed a statistically significant difference between the FM and N-FM diets (p<0.05). After the two weeks period, animals were sacrificed and blood samples obtained for blood chemistry and lipid profile tests. The livers were harvested for antioxidant activity and gene expression assay. The antioxidant assay showed no statistically significant difference among all groups, as well as the blood chemistry and lipid profile. The gene expression assay carried out using two-step Real-time qPCR, showed that PFK-1 gene was more expressed in the DFM group when compared to the DNM and DCG groups. Conclusion: The FM diet enhanced the expression of PFK-1 gene in streptozotocinized-diabetic rats.


2021 ◽  
Vol 12 ◽  
Author(s):  
Etimad A. Huwait ◽  
Salma Y. Saddeek ◽  
Rehab F. Al-Massabi ◽  
Sanaa J. Almowallad ◽  
Peter Natesan Pushparaj ◽  
...  

Background: Atherosclerosis (AS), a major risk factor for stroke and brain tissue destruction, is an inflammatory disease of the blood vessels, and the underlying pathology is inflammation mediated by various chemokines and cytokines. Quercetin, a natural flavonol, is reported to have both anti-inflammatory and antioxidant properties. As such, in the present study, we evaluated the antiatherogenic effects of quercetin in a human THP-1 cell line in vitro and also the signaling mechanisms using in silico analysis.Materials and Methods: THP-1 macrophages exposed to different concentrations of quercetin (5–100 μM for 24 h) were tested for cytotoxicity. Real-time gene expression assay for intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) was carried out following treatment with quercetin at 15 and 30 μM for 24 h either in the absence or presence of interferon (IFN-γ) for 3 h to induce inflammation. Monocyte migration and cholesterol efflux were also assessed.Results: Quercetin did not exert any cytotoxic effects on THP-1 cells at the various concentrations tested. The gene expression assay showed a significant decrease in ICAM-1 (by 3.05 and 2.70) and MCP-1 (by 22.71 and 27.03), respectively. Quercetin at 15 µM decreased THP-1 monocyte migration by 33% compared to the MCP-1-treated cells. It also increased cholesterol efflux significantly by1.64-fold and 1.60-fold either alone or in combination with IFN-γ, respectively. Ingenuity Pathway Analysis of the molecular interactions of quercetin identified canonical pathways directly related to lipid uptake and cholesterol efflux. Furthermore, CD36, SR-A, and LXR-α also demonstrated significant increases by 72.16-, 149.10-, and 29.68-fold, respectively.Conclusion: Our results from both in vitro and in silico studies identified that quercetin inhibited the THP-1 monocyte migration, MCP-1, and ICAM-1 and increased cholesterol efflux probably mediated via the LXR/RXR signaling pathway. Therefore, quercetin will help prevent cell infiltration in atherosclerotic plaques and reduce the risk of stroke or brain destruction.


1970 ◽  
Vol 19 (1) ◽  
pp. 9-23 ◽  
Author(s):  
M. A. Y. Akhond ◽  
G. C. Machray

Optimization of direct DNA delivery into tobacco ((Nicotiana tabacum var. Xanthi) male gametophytes was devised together with development of an efficient transient expression system to study gene expression under controlled conditions. Use of a GFP gene driven by strong promoter and enhancer sequences allowed an efficient non-lethal transient gene expression assay with an overall transient gene expression frequency of > 4% for uninucleate microspores and between 10 and 20% for binucleate pollen. The technique demonstrated its suitability for analysis of developmental stage-specific gene expression. The assay allowed observation of real-time transgene expression during microspore maturation proving useful for in vitro pollen selection. We have also used this protocol to determine the recombination potential of tobacco male gametic cells by assessing the frequency of extra-chromosomal homologous recombination events after co-delivery of two loss-of-function GFP genes. No increase of extrachromosomal recombination was observed in assays for transient transformation. Key words: Biolistic, GFP, Microspore, Tobacco, Nicotiana tabacum, Transformation D.O.I. 10.3329/ptcb.v19i1.4078 Plant Tissue Cult. & Biotech. 19(1): 9-23, 2009 (June)


Planta Medica ◽  
2011 ◽  
Vol 77 (12) ◽  
Author(s):  
M Onrubia ◽  
A Gallego ◽  
K Ramírez ◽  
HR Vidal Limon ◽  
RM Cusidó ◽  
...  
Keyword(s):  

2008 ◽  
Vol 46 (01) ◽  
Author(s):  
F Moriconi ◽  
H Christiansen ◽  
H Christiansen ◽  
N Sheikh ◽  
J Dudas ◽  
...  

2020 ◽  
Vol 139 ◽  
pp. 153-160
Author(s):  
S Peeralil ◽  
TC Joseph ◽  
V Murugadas ◽  
PG Akhilnath ◽  
VN Sreejith ◽  
...  

Luminescent Vibrio harveyi is common in sea and estuarine waters. It produces several virulence factors and negatively affects larval penaeid shrimp in hatcheries, resulting in severe economic losses to shrimp aquaculture. Although V. harveyi is an important pathogen of shrimp, its pathogenicity mechanisms have yet to be completely elucidated. In the present study, isolates of V. harveyi were isolated and characterized from diseased Penaeus monodon postlarvae from hatcheries in Kerala, India, from September to December 2016. All 23 tested isolates were positive for lipase, phospholipase, caseinase, gelatinase and chitinase activity, and 3 of the isolates (MFB32, MFB71 and MFB68) showed potential for significant biofilm formation. Based on the presence of virulence genes, the isolates of V. harveyi were grouped into 6 genotypes, predominated by vhpA+ flaB+ ser+ vhh1- luxR+ vopD- vcrD+ vscN-. One isolate from each genotype was randomly selected for in vivo virulence experiments, and the LD50 ranged from 1.7 ± 0.5 × 103 to 4.1 ± 0.1 × 105 CFU ml-1. The expression of genes during the infection in postlarvae was high in 2 of the isolates (MFB12 and MFB32), consistent with the result of the challenge test. However, in MFB19, even though all genes tested were present, their expression level was very low and likely contributed to its lack of virulence. Because of the significant variation in gene expression, the presence of virulence genes alone cannot be used as a marker for pathogenicity of V. harveyi.


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