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Abstract Accurate measurement of gene expression levels is vital for advancing plant biology research. This study explores the identification and validation of stable reference genes (RGs) for gene expression analysis in Spinacia oleracea. Leveraging transcriptome data from various developmental stages, we employed rigorous statistical analyses to identify potential RGs. A total of 1196 candidate genes were initially screened based on expression variability, with subsequent refinement using criteria such as low variance and stability. Among 12 commonly used candidate RGs, EF1α and H3 emerged as the most stable across diverse experimental conditions, while GRP and PPR exhibited lower stability. These findings were further validated through qRT-PCR assays and comprehensive statistical analyses, including geNorm, NormFinder, BestKeeper, and RefFinder. Our study underscores the importance of systematic RG selection to ensure accurate normalization in gene expression studies, particularly in the context of S. oleracea developmental stages and physiological processes like flowering. These validated RGs provide a robust foundation for future gene expression analysis in S. oleracea and contribute to the advancement of molecular research in plant biology.

Gastrointestinal diseases have been among the main concerns of medical and scientific societies for a long time. Several studies have emphasized the critical role of oxidative stress in the pathogenesis of the most common gastrointestinal diseases. To provide a comprehensive overview of gastrointestinal diseases caused by oxidative stress, their biological aspects, molecular mechanisms and specific pathways, the results of the most recent published articles from the online databases were studied considering both the upper and lower parts of the digestive tract. The results revealed that although the oxidative stress in each part of the digestive system manifests itself in a specific way, all these diseases arise from the imbalance between the generation of the reactive intermediates (especially reactive oxygen species) and the antioxidant defense system. Annual incidence and mortality statistics of gastrointestinal diseases worldwide emphasize the urgent need to find an effective and non-invasive treatment method to overcome these life-threatening problems. Therefore, in the next step, a variety of nanomedicurfines developed to treat these diseases and their effect mechanisms were investigated precisely. Furthermore, the most important nanomedicines responsive to endogenous and exogenous stimuli were evaluated in detail. This review could pave the way to open a new horizon in effectively treating gastrointestinal diseases.

The aim of this work was the simultaneous loading of quercetin and mint essential oil (mint oil) in phospholipid vesicles specifically tailored to obtain an antibacterial and antioxidant mouthwash. The vesicles were prepared using soy lecithin and Tween 80 as bilayer components, and a mixture of phosphate buffer solution (33%), propylene glycol (33%) and ethanol (33%) as dispersing phase. The formation of regularly shaped, spherical and unilamellar vesicles was confirmed by cryogenic transmission electron microscopy analyses. Similarly, light scattering results disclosed that the size of the vesicles increased by increasing the concentration of mint oil, but at the same time the high amount of mint oil ensured high stability, as the size of these vesicles remained unchanged during 12 months of storage. All tested formulations were highly biocompatible towards epithelial cells and capable of counteracting oxidative cell damages caused by hydrogen peroxide. Moreover, the vesicles prepared with the highest concentration of mint oil inhibited the proliferation of the cariogenic Streptococcus mutans (S. mutans) and Lactobacillus acidophilus (L. acidophilus).

Purpose: To potentially enhance the bioavailability and extend the bioactivity effectiveness of Isoleucine-Proline-Proline (IPP, an antihypertensive bioactive peptide of dairy origin), a novel Lyotropic Liquid Crystalline Pharmacosomal Nanoparticle (LLCPNP) was synthesized, and its physicochemical and technological characteristics were studied. Methods: LLCPNPs precursors were developed using IPP and soy phosphatidylcholine via complex formation. Polarized light microscopy, small angle X-ray scattering, differential scanning calorimetry, dynamic light scattering and Fourier transform infrared spectroscopy were employed to characterize the physicochemical properties of the nanoparticles. The in-vitro release and its related mechanisms were also studied. Results: Fourier transform infrared spectroscopy confirmed the complexation between the components of LLCPNPs. Phase behavior evaluation by polarized light microscope showed the characteristic birefringent texture. These findings along with those of small angle X-ray scattering and differential scanning calorimetry proved the formation of lamellar LLCPNPs. These particles represented nanometric size (

Background and Objective: Vocal changes are one of the most common consequences of thyroid surgery. These changes can be due to neurological or other factors. With respect to vocal changes after surgery, and the need for determination of details of the vocal problems and related consequences in the patients’ life after thyroidectomy, the aim of this study was determination of some acoustic parameters of their voice after thyroidectomy and the status of subscales of Voice Handicap Index (VHI). Methods: This study is a cross-sectional descriptive analytic and non-interventional. One voice of 32 thyroidectomized patients (23 women and 9 men) with 20-60 years of age analyzed acoustically and they completed VHI questionnaire. Results: Statistical comparison of mean scores of VHI in two groups of patients with and without vocal problem showed that mean score and standard deviation of total scores of VHI, and the scores of functional, emotional, and physical subscales in patients with voice problems are more than the other group, and the differences were significant (P=0.000). Comparison of acoustic parameters in the two groups of patients showed that F0, jitter, shimmer, and HNR were different, but the differences were not significant. According to our results, amplitude perturbations and harmonic to noise ratio was high in both groups of thyroidectomized patients. Conclusion: According to the findings of this research, we can conclude that F0, jitter, shimmer, and HNR of voice may be not enough for determination of voice problems, but other acoustic measures, self-perceptual, or self-reported evaluation tools such as VHI maybe more efficient in determination of thyroidectomized patients’ voice problems.

Background and Aims: Estimating the canal length is essential for reaching to a proper endodontic treatment. Any error and miscalculation in estimation of the working length, especially in curved canals, can result in complication during and after root canal therapy. The introduction of digital radiography has enabled us to measure curved canal length. The purpose of this study was to evaluate the calibration tool of Complementary Metal Oxide Semiconductor (CMOS) digital system in estimation of working length. Materials and Methods: In this in-vitro study, 80 extracted molar teeth were selected and randomly divided into three groups based on angle and radius of canal curvature. A 5 mm orthodontic wire was placed on the lateral surface of the root. Conventional radiographs were taken after insertion of endodontic file in the apical third of root canals. Radiographic images were scanned and imported to the software of CMOS digital system. Two observers estimated the file lengths with and without using calibration tool of digital measurement software. The correlation between observers was evaluated and then data were analyzed using Paired T-test with 95% confidence. Results: Overall agreement between observers was good. There was no significant difference between the mean values of calibrated measurement and true file length in the basis of canal curves (P>0.001). However, there was significant difference between the mean values of uncalibrated measurement and true file length in the basis of canal curvature (P

The present study aimed at developing a new vesicular formulation capable of promoting the protective effect of ascorbic acid and tocopherol against intestinal oxidative stress damage, and their efficacy in intestinal wound healing upon oral administration. A pH-dependent copolymer (Eudragit® L100), a water-soluble prebiotic fibre (Nutriose® FM06), a phospholipid mixture (Lipoid S75), and two natural antioxidants (ascorbic acid and tocopherol) were combined to fabricate eudragit-nutriosomes by a simple, solvent-free procedure. The vesicles were spherical and oligolamellar, with some multicompartment structures in Eudragit-nutriosomes, small in size (~100 nm), with highly negative zeta potential. The effect of Eudragit® and Nutriose® on the stability on storage and in simulated gastrointestinal fluids were confirmed by the Turbiscan® technology and in vitro studies, respectively. Eudragit-nutriosomes exhibited a protective effect against H2O2-induced oxidative stress, and a proliferative effect in Caco-2 cells, as they provided the closure of the scratched area after 96 h of incubation.