In this study, chitosan nanoparticles (CsNPs) were used as nanocarrier for ultrasonicated ethanolic extract of Rosmarinus officinalis (UEERO) as a new nanoformulation against Eimeria tenella. Herein, CsNPs have been synthesized by ionic gelation method at pH 3 (CsNPs3) and pH 5 (CsNPs5), followed by characterization of morphology, size, polydispersity index (PDI), surface charge, and loading efficiency of UEERO. An in vitro sporulation inhibition assay (10, 5, 2.5, 1.25, 0.62, 0.31, 0.15, 0.07, 0.04, 0.02, and 0.01 mg/ml normal saline solution) against E. tenella was conducted. Results showed that free CsNPs and UEERO-CsNPs3/5 were cubic- and spherical-shaped with positive charge and average size of ~ 150.8 nm (314.4 nm) and 151.7 nm (321.1 nm), respectively. The total loading efficiency using UV-vis spectrophotometer, was 80.05 at pH 5 and 64.39% at pH 3. The in vitro sporulation inhibition assay revealed that UEERO, CsNPs3/5, and UEERO-CsNPs3/5 showed a potential inhibitory effect on sporulation (%), distortion in wall (%), and sporocyst abnormality (%) in a dose-dependent manner. Accordingly, the concentration (10 mg/ml) showed the best efficacy after 24 h in UEERO, free CsNPs, and UEERO-CsNPs. Moreover, UEERO-CsNPs3 and UEERO-CsNPs5 had stopped the sporulation (%) after 72 h. Taken all together, UEERO-CsNPs3 and UEERO-CsNPs5 are best effective against E. tenella in a dose-dependent manner in terms of sporulation (%), distortion in wall (%), and sporocysts abnormality.

BACKGROUND: FOXD1 expression in oral squamous cell carcinoma remains uncovered. The aim was to detect the anticancer effect of Rosemary Extract RE through the evaluation of FOXD1 gene expression in (OSCC) by quantitative PCR.
METHODS: OSCC cell line was served as a control group. Moreover, the OSCC cell line (SCC-15) was treated with RE (OSCC/ RE group) at 24, 48, and 72 hs time intervals. We assessed the antioxidant activity of RE by evaluation of lipid peroxidation (MDA) and superoxide dismutase (SOD) levels. The cytotoxic effects of RE were examined by MTT assay. mTOR and LC3 I/II autophagy protein markers were assessed by western blot. Apoptosis activity was assessed.
RESULTS: The study results were statistically assessed. Intergroup comparisons were analyzed, whereas intragroup comparisons were conducted utilizing one-way repeated measures ANOVA, followed by multiple pairwise paired t-tests with Bonferroni correction revealed a significant increase of FOXD1 gene expression in the control OSCC group in comparison to the OSCC/RE group (p-value <0.001). A significant decrease of mTOR/LC3I/II proteins expression in the OSCC/RE group compared to the control OSCC group (p-value <0.001).
CONCLUSIONS: FOXD1 can be considred a diagnostic biomarker for OSCC. RE inhibits autophagy of oral human cancer cells via mTOR/LC3I/II-dependent pathways and decrease caspase -3 apoptotic level.

In 2017, JECFA requested reproductive and developmental toxicity studies to finalize an acceptable daily intake for solvent rosemary extracts. Thus, an OECD 421 reproductive/developmental toxicity study was conducted using an acetone rosemary extract that complied with JECFA and EFSA food additive specifications. Rosemary extract was provided to rats at dietary concentrations of 0 (control), 2100, 3600, or 5000 mg/kg, for 14 days before mating, during mating, and thereafter (throughout gestation and up to Lactation Day 13 for females) until necropsy. General toxicity (clinical signs, body weight, food consumption) and reproductive/developmental outcomes (fertility and mating performance, estrous cycles, anogenital distance, thyroid hormones, reproductive organ weights, thyroid histopathology) were assessed. There were no signs of general toxicity and no effects on reproduction; thus, the highest concentration tested (equivalent to mean daily intakes of 316 or 401 mg/kg bw/day [149 or 189 mg/kg bw/day carnosol and carnosic acid] for males and females, respectively) was established as the no-observed-adverse-effect level for general and reproductive toxicity. Dose-related reductions in T4 were observed for Day 13 pups (not seen on Day 4) but were not accompanied by thyroid weight changes or histopathological findings; further investigations are required to determine the biological relevance of these T4 reductions.

The antiproliferative activity of Rosemary (Rosmarinus officinalis) has been widely studied in different in vitro and in vivo models, which demonstrate that rosemary extracts inhibit the cellular proliferation due to its ability to interact with a wide spectrum of molecular targets. However, a comprehensive proteomics study in vivo has not been carried out yet. In the present work, the effects of rosemary extract on xenograft tumor growth has been studied and, for the first time, a shotgun proteomic analysis based on nano-LC-MS/MS together with stable isotope dimethyl labeling (DML) has been applied to investigate the global protein changes in vivo. Our results show that the daily administration of a polyphenol-enriched rosemary extract reduces the progression of colorectal cancer in vivo with the subsequent deregulation of 74 proteins. The bioinformatic analysis of these proteins indicates that the rosemary extract mainly alters the RNA Post-Transcriptional Modification, the Protein Synthesis and the Amino Acid Metabolism functions and suggests the inactivation of the oncogene MYC. These results demonstrate the high utility of the proposed analytical methodology to determine, simultaneously, the expression levels of a large number of protein biomarkers and to generate new hypothesis about the molecular mechanisms of this extract in vivo.

In this work, a proteomics strategy based on nanoliquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) using an Orbitrap high-resolution mass spectrometer together with stable isotope dimethyl labeling (DML) is applied to quantitatively examine relative changes in the protein fraction of HT-29 human colon cancer cells treated with different concentrations of a polyphenol-enriched rosemary extract over the time. The major objective of this study was to gain insights into the antiproliferative mechanisms induced by rosemary polyphenols. Using this methodology, 1909 and 698 proteins were identified and quantified in cell extracts. The polyphenol-enriched rosemary extract treatment changed the expression of several proteins in a time- and concentration-dependent manner. Most of the altered proteins are implicated in the activation of Nrf2 transcription factor and the unfolded protein response. In conclusion, rosemary polyphenols induced proteomic changes that were related to the attenuation of aggresome formation and activation of autophagy to alleviate cellular stress.

There is a tendency towards the use of natural antioxidative substances due to toxic effects of synthetic antioxidants. The purpose of this research was to evaluate the effects of tocopherol and extracts from rosemary and ferulago on oxidation in a mixture (1:1, w/w) of sunflower seed oil and palm olein, during deep frying of potato slices. Besides the control groups, tertiary butylhydroquinone (TBHQ) served as a standard for comparison. The DPPH radical scavenging activities of extracts followed the order of rosemary10% > rosemary1% > rosemary0.1 % > ferulago 10 % > ferulago 1 % > ferulago 0.1 %. Frying performance of antioxidants were tested with regards to primary (peroxide value (PV)) and secondary (anisidine value (AnV), free fatty acids (FFA), total polar compounds (TPC) and volatile oxidation compounds such as hexanal and heptanal) oxidation products and by sensory evaluation. After frying process, rosemary extract treatment exhibited PV (1.2 meq O2/kg), FFA content (0.124 %), TPC (12.2 %), hexanal concentration (62.4 ng/g) and heptanal concentration (73.8 ng/g), which were significantly equal or sometimes less than those of TBHQ treatment ((1.23 meq O2/kg), (0.123 %), (12.6 %), (64.4 ng/g) and (74.1 ng/g), respectively). Whilst the ferulago extract showed higher PV (2 meq O2/kg) than the control sample, it was efficient in delaying secondary oxidative changes. Tocopherol markedly retarded primary stage but its activity fell down toward secondary oxidation products. The overall results of this research suggest that rosemary extract may be a good natural alternative to TBHQ, but further studies are necessary to investigate the use of ferulago extract.

A comparative study on the antioxidant effect of rosemary extract (RE) and sodium ascorbate (SA) on lipid and colour oxidation of liver pâté made of lard and pork liver was done. During the 48 hour experimental time all the pâtés were wrapped in a foil and stored in cold room of 3.5°C under light of 1000 lux. Colour stability was monitored by instrumental colour measurement (CIE L*a*b* colour space) whereas lipid stability was measured by the determination of the 2-thiobarbituric acid reactive substances (TBARS). In the present study RE doses range (0,125, 250, 375 and 500 ppm) showed no significant (p>0.05) and linear effect on colour stability. However thez RE revealed a significant effect (p<0.05) against lipid oxidation and linearly reduces the TBARS number. The added SA doses (0, 250, 500, 750 and 1000 ppm) revealed significant (p<0.05) and linear effect in reducing discoloration. However the studied SA dose ranges showed no significant (p>0.05) effect on TBARS number. In this study RE was showed better performance against lipid oxidation and SA was potent against discoloration. The effect of the added spices used in manufacturing of the studied product showed no significant (p>0.05) effect against lipid and color oxidation. However the added spices revealed possible antagonistic and synergetic relationship with the studied the antioxidant systems (RE & SA).