Mexican oregano is composed by a wide range of endemic species which support important economic activities, but with serious limitations. In this work a micropropagation protocol was established to regenerate oregano plants (Poliomintha glabrescens Gray) from apical buds. At the same time we evaluated the impact of this process in the phytochemical profile of the new plants. The optimal induction of apical buds was observed when shoottip explants were incubated on solidified Murashige and Skoog (MS) medium supplemented with 0.5 mg ml -1 anaphthalenacetic acid (NAA) and 0.02 mg ml -1 6-benzyladenine. Also, the optimal response to root induction was with MS solid medium added with 0.5 mg ml -1 of NAA. The micropropagation protocol took 17 weeks with an efficiency of 40%, and plants achieved were successfully acclimatized under greenhouse conditions. The total phenolics (TP) concentration, antioxidant capacity (oxygen radical absorbance capacity, ORAC), and luteolin content from methanol extracts of wild type (WT) and in vitro (IN) and ex vitro (EX) plants were determined. The results show that TP content and ORAC were similar for WT and EX plants, but IN plants had the highest TP content and ORAC activity (25 and 28% respectively). Luteolin content showed significant differences between WT, IN and EX, with EX plants having the highest luteolin content (10% more compared to WT). In summary, we successfully microprogated oregano plants from WT and they contained similar amount of their TP content and ORAC activity and an increase in luteolin content.
Phenolic extracts obtained from spices are known to have anti-carcinogenic activities but little is known about the effect of micropropagation on these beneficial effects. The main objective of this study was to evaluate the cytotoxic activity of flavonoid-enriched extracts (FEE) from the leaves of wild (WT), in vitro (IN), and ex vitro (EX) grown oregano plants in colon cancer cells HT-29 and the non-cancer cells CCD-18Co. Cell proliferation of HT-29 cells was reduced to 50 % by WT, IN, and EX at concentrations of 4.01, 1.32, and 4.84 mg of gallic acid equivalents (GAE)/L, respectively. In contrast, in CCD-18Co cells, higher concentrations were required for the same cytotoxic effect. At 6 mg GAE/L, WT and IN reduced the production of reactive oxygen species (ROS) of lipopolysaccharides (LPS)-stimulated control cells to 59.89 and 59.43 %, respectively, and EX to 73.89 %. The mRNA of Caspase-3 was increased 1.53-fold when cells were treated with 4 mg GAE/L of IN extract, and tumor necrosis factor receptor superfamily, member 6 (FAS), and BCL2-associated X protein (BAX) mRNA increased 2.55 and 1.53 fold, respectively. Results on protein expression corroborated the apoptotic effects with a significant decrease of B-cell lymphoma 2 (BCL2) expression for all treatments but more remarkable for EX that also showed the most intense signal of BAX. Overall, FEE extracts derived from micropropagation had increased pro-apoptotic effects, however extracts from the in vitro plants produced more efficacy at the transcriptional level while extracts from the ex vitro plant were superior at the traductional level.
Ochratoxin A (OTA) is a mycotoxin frequently found in raw and processed foods. While it is considered a possible human carcinogen, the mechanism of action remains unclear. OTA has been shown to be hepatotoxic in both in vitro and in vivo models and oxidative stress may be one of the factors contributing to its toxicity. Hence, the effect of OTA on human hepatocellular carcinoma, HepG2 cells, was investigated on oxidative stress parameters. The cytotoxicity of OTA on HepG2 was time- and dose-dependent within a range between 0.1 and 10 µM; while 100 μM of OTA increased the intracellular reactive oxygen species (ROS) in a time-dependent manner. Additionally, the levels of glutathione (GSH) were increased by 9.7% and 11.3% at 10 and 100 nM of OTA, respectively; while OTA at 100 μM depleted GSH by 40.5% after 24 h exposure compared with the control. Finally, the mRNA level of catalase (CAT) was downregulated by 2.33-, 1.92-, and 1.82-fold after cells were treated with 1, 10, and 10 μM OTA for 24 h, respectively; which was linked to a decrease in CAT enzymatic activity. These results suggest that oxidative stress is involved in OTA-mediated toxicity in HepG2 cells.
El orégano es una planta de distribución mundial, el cual está representado principalmente por dos especies: Origanum vulgare (Lamiaceace) nativo de Europa, y Lippia graveolens (Verbenaceae), originaria de América. México ocupa el segundo lugar como productor mundial de orégano con la especie L. graveolens H. B. K. Sin embargo, la mayoría de las especies explotadas son silvestres y su cultivo es aún tradicional y limitado. En este trabajo se presenta un análisis de las estrategias de propagación, composición química y propiedades nutracéuticas del orégano. Los fitoquímicos presentes pueden clasificarse en tres categorías: compuestos volátiles, lípidos y fenólicos. Estos componentes presentan diversas propiedades nutracéuticas entre las que destacan la actividad antioxidante, hipoglucémica, hipotensiva, hipolipidémica y anticancerígena. Los avances en el estudio de la composición fitoquímica y su relación con nuevas propiedades nutracéuticas hacen del orégano un producto de alto valor comercial con amplias aplicaciones quimioterapéuticas.
This report describes a disorder of the sexual development in a beagle dog resulting in an intersex condition. A 6 mo old beagle was presented for evaluation of a protruding structure from the vulva consistent with an enlarged clitoris. Ultrasonographic examination revealed the presence of both gonadal and uterine structures. Retrograde cystourethrovaginogram showed the presence of an os clitoris and severe vaginal stenosis. Histological studies revealed the presence of bilateral ovotestes and uterus. The gonad had interstitial cells within seminiferous-like tubules lined only with Sertoli cells and abundant interstitial cells among primordial, primary, and secondary follicles. Hormone assays completed before and after gonadohysterectomy showed an elevation in the levels of progesterone and dihydrotestosterone that returned to baseline 3 mo after surgery. Testosterone levels that were within the male reference ranges before surgery decreased to basal levels postsurgically. 17-β-Estradiol levels showed little variation and values were always within the reference ranges for a male. Cytogenetic analysis showed a normal female karyotype (2n = 78, XX) and polymerase chain reaction analysis revealed the absence of the sex-determining region Y gene. In summary, the dog presented bilateral ovotestes and a 2n = 78, XX chromosomal complement lacking the sex determining region Y gene, consistent with a diagnosis of true hermaphroditism.
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