RESUMO
The human microbiome is a complex network of microorganisms that includes viruses, bacteria, and fungi. The gut virome is an essential component of the immune system, which is responsible for regulating the growth and responses of the host's immune system. The virome maintains a crucial role in the development of numerous diseases, including inflammatory bowel disease (IBD), Crohn's disease, and neurodegenerative disorders. The human virome has emerged as a promising biomarker and therapeutic target. This comprehensive review summarizes the present understanding of the virome and its implications in matters of health and disease, with a focus on the Human Microbiome Project.
Assuntos
Microbioma Gastrointestinal , Viroma , Humanos , Doenças Inflamatórias Intestinais/virologiaRESUMO
The presence of salt can impact the fluid phase and gelatinization process of starch granules. The variation in viscosity and rheology models including the Herschel-Bulkley, the Casson model, and the power law, were determined by adding salts before and after starch ultrasonication. Non-isothermal kinetics can be utilized for the mathematical modeling of the gelatinization process and the evolution of the reaction. Unlike Na+ ions, Ca+2 ions notably elevate viscosity. The Casson model accurately predicts viscosity data. Results indicate that the addition of Na+ ions decreases yield stress by up to 60.4 %, while Ca+2 ions increase by up to 100.8 %. Adding Na+ ions decreases the required thermal energy by as much as 49.6 %, while the presence of Ca+2 ions can lead to a substantial increase of up to 337.1 % compared to control samples. The positive ∆G indicates a non-spontaneous gelatinization process. The addition of NaCl promotes a spontaneous reaction, while the addition of CaCl2 increases the Gibbs energy. The changes in entropy are minimal, implying minimal changes in starches' disorder structure.
Assuntos
Cloreto de Cálcio , Manihot , Reologia , Cloreto de Sódio , Amido , Termodinâmica , Amido/química , Cinética , Cloreto de Sódio/química , Cloreto de Cálcio/química , Manihot/química , Viscosidade , Gelatina/químicaRESUMO
Present study investigates the effects of probe size geometry on thermodynamic kinetics, rheology, and microstructure of wheat and tapioca starch. Ultrasound treatment using different probe diameters (20 mm and 100 mm) significantly influenced the gelatinization process. Results showed reduced enthalpy (ΔH) and Gibbs energy (ΔG), indicating enhanced gelatinization efficiency. According to the results, using a 20 mm and 100 mm probe leads to a reduction of 52.7 % and 68.6 % in reaction enthalpy for wheat starch compared to native starch, respectively. Microstructure analysis revealed structural changes, with ultrasound treatment leading to granular fractures and a sheet-like structure with air bubbles. The rheological behavior of the starches is found to exhibit shear thinning behavior, with the Casson model providing the best fit for the experimental data. Moreover, rheology modeling using Herschel-Bulkley and power law models showed increased viscosity and shear stress in larger probes. Numerical simulation data demonstrated that probe size influenced ultrasonic pressure, sound pressure level, and thermal power dissipation density, affecting fluid motion and velocity field components. Moreover, the maximum dissipated power decreases from 8.43 to 0.655 mW/m3 with an increase in probe diameter from 20 to 100 mm. The average yield shear stress values are calculated as 3.36 and 3.14 for wheat and tapioca starches, respectively. The larger probe diameter leads to greater entropy increases, with tapioca starch showing a 4.72 % increase and wheat starch a 4.97 % increase, compared to 2.56 % and 3.11 %, respectively, with the smaller probe. Additionally, the Keller-Miksis model provided insights into bubble dynamics, revealing increased pressure and temperature with higher pressure amplitudes.
RESUMO
Diabetes is a global problem that threatens human health. Cell therapy methods using stem cells, and tissue engineering of pancreatic islets as new therapeutic approaches have increased the chances of successful diabetes treatment. In this study, to differentiate Wharton's Jelly-derived mesenchymal stem cells (WJ-MSCs) into insulin-producing cells (IPCs) with improved maturity, and function, platelet-rich plasma (PRP)-Polyvinylpyrrolidone (PVP)-Polycaprolactone (PCL)/PCL scaffold was designed. The two-dimensional (2D) control group included cell culture without differentiation medium, and the experimental groups included 2D, and three-dimensional (3D) groups with pancreatic beta cell differentiation medium. WJ-MSCs-derived IPCs on PRP-PVP-PCL/PCL scaffold took round cluster morphology, the typical pancreatic islets morphology. Real-time PCR, immunocytochemistry, and flowcytometry data showed a significant increase in pancreatic marker genes in WJ-MSCs-derived IPCs on the PRP-PVP-PCL/PCL scaffold compared to the 2D-experimental group. Also, using the ELISA assay, a significant increase in the secretion of insulin, and C-peptide was measured in the WJ-MSCs-derived IPCs of the 3D-experimental group compared to the 2D experimental group, the highest amount of insulin (38 µlU/ml), and C-peptide (43 pmol/l) secretion was in the 3D experimental group, and in response to 25 mM glucose solution, which indicated a significant improvement in the functional level of the WJ-MSCs-derived IPCs in the 3D group. The results showed that the PRP-PVP-PCL/PCL scaffold can provide an appropriate microenvironment for the engineering of pancreatic islets, and the generation of IPCs.
Assuntos
Diabetes Mellitus , Células-Tronco Mesenquimais , Poliésteres , Povidona/análogos & derivados , Geleia de Wharton , Humanos , Peptídeo C , Diferenciação Celular , Células CultivadasRESUMO
In this study, thermosonication (37 KHz, 300 W; 50, 60, and 70 °C) of celery juice was performed to inactivate Escherichia coli and Salmonella Typhi in 6 min. The inactivation of pathogens and the process were modeled using mathematical, thermodynamic, and computational fluid dynamics models. The findings indicated that the distribution of power dissipation density was not uniform across the entire domain, including the beaker area, with a maximum value of 27.8 × 103 W/m3. At lower temperatures, E. coli showed a 9.4 % higher resistance to sonication, while at higher temperatures, S. Typhi had a 5.4 % higher durability than E. coli. Increasing the temperature decreased the maximum inactivation rate of both S. Typhi and E. coli by 15.5 % and 20.5 % respectively, while increasing the thermal level by 20 °C reduced the log time to achieve the maximum inactivation rate by 20.3 % and 34.9 % for S. Typhi and E. coli respectively, highlighting the stronger effect of sonication at higher temperatures. According to the results, the positive magnitudes of ΔG were observed in both E. coli and S. Typhi, indicating a similar range of variations. Additionally, the magnitude of ΔG increased by approximately 5.2 to 5.5 % for both microorganisms which suggested the inactivation process was not spontaneous.
Assuntos
Apium , Escherichia coli , Salmonella typhi , Hidrodinâmica , Bebidas , TemperaturaRESUMO
In this study, the effects of ultrasonicated Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus and Lactiplantibacillus plantarum AF1 (100 W, 30 kHz, 3 min) on the safety and bioactive properties of stirred yoghurt during storage (4 °C for 21 days) were investigated. The results showed that sonicated cultures were more effective in reducing pathogens than untreated ones. The highest antioxidant activity (DPPH and ABTS), α-glucosidase and α-amylase inhibition capacity were found in yoghurt containing sonicated probiotic + sonicated yoghurt starter cultures (P + Y + ). The highest amount of peptides (12.4 mg/g) was found in P + Y + yoghurts at the end of the storage time. There were not significant differences between the exopolysaccharide content of P + Y+ (17.30 mg/L) and P + Y- (17.20 mg/L) yoghurts, although it was significantly (P ≤ 0.05) higher than the other samples. The use of ultrasonicated cultures could enhance the safety of stirred yoghurt and improve its functional and bioactive properties.
Assuntos
Lactobacillus delbrueckii , Lactobacillus plantarum , Lactobacillus delbrueckii/fisiologia , Streptococcus thermophilus , Iogurte , FermentaçãoRESUMO
To achieve wheat starch acetylation (AC) with a high degree of substitution (DS), the acetylation process was carried out using various ultrasonication frequencies, including 25 kHz, 40 kHz, and 25 + 40 kHz. In the second step, wheat starch's ultrasound-assisted acetylation (UAA) is simulated using various approaches including the rheology models, non-isothermal reaction kinetics, and flow/acoustic modelling. The computational fluid dynamics (CFD) simulation solves the non-linear acoustic governing equation to determine the flow field and the amount of delivered ultrasound energy. The acetylated starch increased peak and final viscosity, with the highest values observed for the 25 + 40 kHz frequency than other single frequencies (25 kHz and 40 kHz). The viscosity of the starch is specified based on the experimental data using Herschel-Bulkley, power law, and Casson rheology models. According to differential scanning calorimetry (DSC) analysis, the gelatinization parameters and enthalpy of gelatinization (ΔHgel), were found to be lower in acetylated starches at the frequency of 25 + 40 kHz compared to those at frequencies of 25 kHz and 40 kHz, as well as native starches (NS). Moreover, the gelatinization process is examined by implementing the non-isothermal reaction kinetics to obtain the activation energy and reaction order. Based on the results obtained, implementing sonication at 25 kHz reduces the activation energy by 70.3 % compared to native starch. However, the same parameter is obtained to be 69.9 % and 67.1 % for the application of 40 and 25 + 40 kHz transducers, respectively. Additionally, during the sonication treatment, the yield shear stress increases between 24.1 and 31.8 %, based on the applied frequency. Morphology analysis determined by scanning electron microscopy (SEM) revealed that the surfaces and small granules underwent more damage in acetylated starches at frequencies of 25 kHz and 40 kHz. However, in acetylated starches at 25 + 40 kHz, the larger granules were more affected than the smaller ones.
RESUMO
Epstein-Barr virus-related malignancies have been linked to variations in the sequences of EBV genes, notably EBNA1. Therefore, the purpose of this study was to examine the DBD/DD domain and USP7 binding domain sequences at the C-terminus of the EBNA1 gene in patients with chronic lymphocytic leukemia (CLL). This study included 40 CLL patients and 21 healthy volunteers. Using commercial kits, total DNA was extracted from buffy coat samples, and each sample was tested for the presence of the EBV genome. The C-terminus of EBNA1 was then amplified from positive samples, using nested PCR. Sanger sequencing was used to identify mutations in the PCR products, and the results were analyzed using MEGA11 software. The mean ages of CLL patients and healthy individuals were 61.07 ± 10.2 and 59.08 ± 10.3, respectively. In the EBNA-1 amplicons from CLL patients and healthy individuals, 38.5% and 16.7%, respectively, harbored mutations in the DBD/DD domain of the C-terminal region of the EBNA1 gene (P = 0.378). The mutation frequency at locus 97,320 was significantly higher in CLL patients than in healthy individuals (P = 0.039). Three EBV subtypes based on residue 487 were detected. The frequency of alanine, threonine, and valine in both groups was 88, 8, and 4 percent, respectively (P = 0.207). Moreover, all of the isolates from healthy donors had alanine at this position. The findings indicated that the presence of threonine or valine at residue 487 as well as a synonymous substitution at residue 553 in the C-terminal region of EBNA1 might be involved in the pathogenesis of EBV in CLL patients.
Assuntos
Infecções por Vírus Epstein-Barr , Antígenos Nucleares do Vírus Epstein-Barr , Leucemia Linfocítica Crônica de Células B , Humanos , Alanina , Antígenos Nucleares do Vírus Epstein-Barr/genética , Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Voluntários Saudáveis , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/metabolismo , Leucemia Linfocítica Crônica de Células B/genética , Leucemia Linfocítica Crônica de Células B/virologia , Treonina , Peptidase 7 Específica de Ubiquitina , ValinaRESUMO
Background: As infection with rubella and varicella-zoster viruses (VZV) can lead to congenital syndrome and its dangerous complications, assessing immunity to these congenital infections can represent the biological risk assessment related to their exposure in high-risk groups. Therefore, we aimed to evaluate the frequency of IgG anti-varicella/rubella antibodies (Abs) in female students at Shiraz University of Medical Sciences (SUMS), Iran. Patients and Methods: In this study, a total of 434 female students were included. Sera were isolated from blood samples and stored at -20°C for later analysis. A questionnaire form was documented and contained demographic data as well as the history of vaccinations. Enrolled students were divided into recipients of either one or two doses of the measles/rubella (MR) vaccine. Serum samples were analyzed for rubella and VZV IgG Abs using commercial IgG immunoassays. Results: The students were 21.6±4.25 years old on average. Out of the 434 enrolled students, 292 (67.3%) and 287 (66.1%) students were positive for anti-varicella and anti-rubella IgG-Abs, respectively. The frequency of anti-rubella IgG Ab was significantly higher in those who received one dose of MR vaccine (P<0.001). In addition, 205 (47.2%) and 59 (13.6%) students were double-positive (anti-varicella/rubella IgG Abs) and double-negative, respectively. Conclusion: Our results indicated that an additional dose of rubella vaccine may be required for those who received two doses of the vaccine. In addition, we recommend the inclusion of the VZV vaccine in Iran's routine vaccination program. Further studies are recommended to verify these results.
RESUMO
In the present research, the impact of fermentation with two strains of Lactiplantibacillus plantarum subsp. plantarum (PTCC 1896 and PTCC 1745) on physicochemical properties, antioxidant bioactive compounds, and some health-promoting features of guava juice was investigated. Results showed a significant (p < .05) decrease in pH, total soluble solids, glucose and fructose residues, vitamin C, and total carotenoids after 32 h of fermentation. Total phenolic content, free radical scavenging abilities, and ferrous reducing power were markedly enhanced during the fermentation process. Moreover, fermented juice represented good enzyme inhibition abilities (α-amylase and α-glucosidase) and anti-inflammatory activities. The initial amount of angiotensin-converting enzyme inhibitory activity (26.5%) increased to 72.1% and 66.4% in L. plantarum subsp. plantarum 1896 and L. plantarum subsp. plantarum 1745 treatments, respectively. These findings reveal that guava juice fermentation with the studied Lactobacillus strains can be a promising strategy to augment the functional properties of the fruit-based beverage.
RESUMO
Wheat is the most important crops worldwide, providing about one-fifth of the daily protein and calories for human consumption. The quality of cereal-based products is principally governed by the genetic basis of gluten (glutenin and gliadin proteins), which exists in a wide range of variable alleles and is controlled by clusters of genes. There are certain limitations associated with gluten characteristics, which can be genetically manipulated. The present review aimed to investigate the correlation between the genetic characteristics of gluten protein components and wheat-based product's quality. According to various references, Glu-B1d (6 + 8), Glu-B1h (14 + 15) and Glu-B1b (7 + 8) are related to higher gluten strength and pasta quality, while, subunits Dx2 + Dy12 and Dx5 + Dy10, are usually present at the Glu-D1 locus in bread wheat, resulted in lower cooked firmness in pasta. Moreover, introducing Gli-D1/Glu-D3 and Glu-D1 loci into durum wheat genomes, causing to provide the maximum values of gluten index in pasta products. 1Dx5 + 1Dy10 alleles determine the level of increase in dough's consistency, elasticity, viscosity, and extensibility quality of baking and appropriate bread loaf volume, while 1Dx2 + 1Dy12 as the alleles associated with poor baking quality, being more suitable for soft wheat/pastry end uses. Bx7, Bx7OE, 1Bx17 + 1By18, 1Bx13 + 1By16, Bx7 + By9 and 1Bx7 + 1By8 at Glu-B1alleles and 1Ax2* found on Glu-A1, augmented dough strength and has positive effects on consistency, extensibility, viscosity, and elasticity of bread dough. Breeding programs by genome editing have made gluten a promoting component for improving cereal-based products.
RESUMO
This study examined the effect of beef fermentation with Lactiplantibacillus paraplantarum (L) PTCC 1965, Lactiplantibacillus (L) plantarum subsp. plantarum PTCC 1745, and Lactiplantibacillus (L) pentosus PTCC 1872 bacteria on the growth of pathogenic bacteria, including Salmonella (S) Typhi PTCC 1609 and Staphylococcus (S) aureus PTCC 1826. The growth of lactic acid bacteria (LAB) and the effect of fermentation on pathogenic bacteria were studied using Weibull: biphasic linear and competitive models. The results showed that the rate of pH reduction was lower in the early stages and increased as the microbial population grew. The α parameter was lower for L. plantarum subsp. plantarum compared to L. paraplantarum and L. pentosus. The comparison of the α parameter for bacterial growth and pH data showed that the time interval required to initiate the rapid growth phase of the bacteria was much shorter than that for the rapid pH reduction phase. The pH value had a 50% greater effect on the inactivation of S. Typhi when compared to the samples containing L. plantarum subsp. plantarum and L. pentosus. The same parameter was reported to be 72% for the inactivation of St. aureus. In general, during the fermentation process, LAB strains caused a decrease in pH, and as a result, reduced the growth of pathogens, which improves consumer health and increases the food safety of fermented meat.
RESUMO
The high water and nutritional contents of meat and fish products make them susceptible to spoilage. Thus, one of the most important challenges faced by the meat industry is extending the shelf life of meat and fish products. In recent years, increasing concerns associated with synthetic compounds on health have limited their application in food formulations. Thus, there is a great need for natural bioactive compounds. Direct use of these compounds in the food industry has faced different obstacles due to their hydrophobic nature, high volatility, and sensitivity to processing and environmental conditions. Nanotechnology is a promising method for overcoming these challenges. Thus, this article aims to review the recent knowledge about the effect of biopolymer-based edible films or coatings on the shelf life of meat and fish products. This study begins by discussing the effect of biopolymer (pectin, alginate, and chitosan) based edible films or coatings on the oxidation stability and microbial growth of meat products. This is followed by an overview of the nano-encapsulation systems (nano-emulsions and nanoliposomes) and the effect of edible films or coatings incorporated with nanosystems on the shelf life of meat and fish products.
RESUMO
Measles is an acute, highly contagious disease with a high mortality rate in children. Although vaccination has reduced measles incidence, outbreaks still occur. Therefore, in this study, we aimed to investigate the frequency of antimeasles immunoglobulin G (IgG) antibody (Ab) among students at Shiraz University of Medical Sciences (SUMS). Four hundred fifty SUMS students were enrolled in this cross-sectional study. Information on demographics and measles vaccination history was collected using a questionnaire. Participants were divided into two groups, including A and B, according to routine doses of measles vaccine and the national measles/rubella immunization program. The antimeasles IgG Abs were tested using a commercial Enzyme-Linked Immunosorbent Assay Kit. Participants ranged in age from 18 to 48 years, with a mean age of 22.2 (±4.3). Fifty percent of the subjects were male. Our results showed that 63.6% of the cases were positive for antimeasles IgG Abs. The seroprevalence of IgG Abs between groups A and B did not differ significantly (p = 0.612). There was also no significant correlation between the seroprevalence of antimeasles IgG Abs and the age (p = 0.43) or sex (p = 0.24) of the subjects. The results showed that the frequency of antimeasles IgG Abs is lower than required to prevent the measles virus from circulating. Therefore, a booster vaccination may be necessary.
Assuntos
Sarampo , Caxumba , Rubéola (Sarampo Alemão) , Criança , Humanos , Masculino , Adulto Jovem , Adulto , Adolescente , Pessoa de Meia-Idade , Feminino , Imunoglobulina G , Estudos Soroepidemiológicos , Estudos Transversais , Anticorpos Antivirais , Sarampo/epidemiologia , Sarampo/prevenção & controle , Rubéola (Sarampo Alemão)/epidemiologia , Estudantes , Vacinação , Vacina contra Sarampo-Caxumba-Rubéola , Caxumba/epidemiologia , Caxumba/prevenção & controleRESUMO
Avian influenza viruses (AIVs) are globally challenging due to widespread circulation and high mortality rates. Highly pathogenic avian influenza (HPAI) strains like H5N1 have caused significant outbreaks in birds. Since 2003 to 14 July 2023, the World Health Organization (WHO) has documented 878 cases of HPAI H5N1 infection in humans and 458 (52.16%) fatalities in 23 countries. Recent outbreaks in wild birds, domestic birds, sea lions, minks, and etc., and the occurrence of genetic variations among HPAI H5N1 strains raise concerns about potential transmission and public health risks. This paper aims to provide a comprehensive overview of the current understanding and new insights into HPAI H5N1. It begins with an introduction to the significance of studying this virus and highlighting the need for updated knowledge. The origin and evaluation of HPAI H5N1 are examined, shedding light on its emergence, and spread across different geographic regions. The genome organization and structural biology of the H5N1 virus are explored, providing insights into its molecular composition and key structural features. This manuscript also delves into the phylogeny, evolution, mutational trends, reservoirs, and transmission routes of HPAI H5N1. The immune response against HPAI H5N1 and its implications for vaccine development are analyzed, along with an exploration of the pathogenesis and clinical manifestations of HPAI H5N1 in human cases. Furthermore, diagnostic tools and preventive and therapeutic strategies are discussed, highlighting the current approaches and potential future directions for better management of the potential pandemic.
RESUMO
Background: Several countries, including Iran, have been affected by the novel Coronavirus Disease 2019 (COVID-19) pandemic since December 2019. The aim of this study was to provide a comprehensive report on COVID-19 patients in Shiraz, Southern Iran. Materials and Methods: This study was performed on 311 hospitalized patients with COVID-19. The data on demographic, clinical, and paraclinical features were analyzed. Results: The median age of the patients was 58 years, with 42.1% of the patients being above 60 years of age. Upon admission, fever was detected in 28.2% of critically ill patients. At least one underlying disease or risk factor was also present in 75.6% of the patients. Shortness of breath was the most common clinical symptom (66.2%), dry cough (53.7%), and muscle pain (40.5%) was the second and third. Sneezing (0.3%), rhinorrhea (0.7%), and sore throat (3.09%) were observed only in non-critically ill patients. In addition, 26.9% of all patients had lymphocytopenia, 25.8% had raised C-reactive protein, and 79.9% had abnormal creatinine levels. Finally, death occurred in 39 patients (12.5%). Conclusions: Noncritically ill patients were younger than critically ill patients. The most common risk factors for getting critically ill were surgery, hypertension, diabetes mellitus, chronic heart disease, asthma, and chronic renal disease.
RESUMO
Background & Objective: Epstein-Barr virus nuclear antigen-1 (EBNA1) is one of the most important proteins of Epstein-Barr virus (EBV) that might be mutated in various related cancers. The purpose of this study was to compare EBNA1 mutations in the C-terminal region between patients with cervical and ovarian cancer and healthy individuals. Methods: As test and control groups, 18 EBV-positive paraffin-embedded samples of cervical and ovarian cancer and 10 age- and gender-matched healthy volunteers who did not have cancer but were EBV-positive were both used. Utilizing a commercial DNA extraction kit, total DNA was extracted following deparaffinization. The entire C-terminal region of the EBNA1 sequence was amplified using an in-house nested PCR. Phylogenetic analysis and Sanger sequencing were used to analyze the sequences using MEGA 7 software and through NJ method. Results: Sequence analysis revealed that the P-Ala subtype of EBNA1 was present in all samples. In two and one samples, respectively, of cervical cancer patients, the mutations A1887G and G1891A were found. The G1595T mutation was also detected in four sequences taken from ovarian cancer patients. No statistically significant difference could be found between the frequency of mutations in patients and controls (P>0.05). No known amino acid substitutions were found in the USP7-binding region and the DBD/DD domain. Conclusion: The findings showed that P-Ala is the predominant EBV subtype across all samples. Additionally, as the sequence of EBNA1's C-terminal region is so stable, it's possible that it had little impact on the pathogenesis of ovarian and cervical malignancies. It is advised to conduct additional research to verify these findings.
RESUMO
This study investigated the effect of Rosmarinus officinalis L. essential oil, REO (one, two and three percent) on the microbiological and oxidative stability of Sarshir during 20 days of refrigerated storage (4 °C). Initially, the chemical composition (gas chromatography/mass spectrometry, GC/MS), antimicrobial (paper disc diffusion) and antioxidant (DPPH) properties of REO were evaluated. Then, the microbial safety, oxidative stability (peroxide and anisidine values) and overall acceptability of the product after addition of REO to Sarshir and the subsequent storage period were determined. According to GC/MS analysis, the major components of REO were α-pinene (24.6%), 1,8-cineole (14.1%), camphor (13.5%), camphene (8.1%) and limonene (6.1%), respectively. Moreover, it was also found that Limosilactobacillus fermentum (inhibition zone (IZ) of 23.5 mm) and Salmonella Typhi (IZ of 16.4 mm) were the most sensitive and resistant spoilage and pathogenic bacteria against REO, respectively. In addition, the half-maximal inhibitory concentration (IC50) of the REO was measured at 24.8 mg/mL, while the IC50 value of butylated hydroxytoluene (BHT) was 16.6 mg/mL. The highest and lowest bacterial populations were detected in the control and the sample containing 3% REO, respectively. The control had the highest extent of lipid oxidation, while the lowest peroxide and anisidine values were measured in Sarshir containing 3% REO.
Assuntos
Óleos Voláteis , Rosmarinus , Rosmarinus/química , Óleos Voláteis/farmacologia , Óleos Voláteis/química , Eucaliptol , Peróxidos , Estresse OxidativoRESUMO
Background: Suppression of p53 is an important mechanism in Epstein-Barr virus associate-tumors and described as EBNA1-USP7 which is a key axis in p53 suppression. Thus, in this study, we aimed to evaluate the function of EBNA1 on the expression of p53-inhibiting genes including HDAC-1, MDM2, MDM4, Sirt-3, and PSMD10 and the influence of USP7 inhibition using GNE-6776 on p53 at protein/mRNA level. Methods: The electroporation method was used to transfect the BL28 cell line with EBNA1. Cells with stable EBNA1 expression were selected by Hygromycin B treatment. The expression of seven genes, including PSMD10, HDAC-1, USP7, MDM2, P53, Sirt-3, and MDM4, was evaluated using a real-time PCR assay. For evaluating the effects of USP7 inhibition, the cells were treated with GNE-6776; after 24 hours and 4 days, the cells were collected and again expression of interest genes was evaluated. Results: MDM2 (P=0.028), MDM4 (P=0.028), USP7 (P=0.028), and HDAC1 (P=0.015) all showed significantly higher expression in EBNA1-harboring cells compared to control plasmid transfected cells, while p53 mRNA expression was only marginally downregulated in EBNA1 harboring cells (P=0.685). Four-day after treatment, none of the studied genes was significantly changed. Also, in the first 24-hour after treatment, mRNA expression of p53 was downregulated (P=0.685), but after 4 days it was upregulated (P=0.7) insignificantly. Conclusion: It seems that EBNA1 could strongly upregulate p53-inhibiting genes including HDAC1, MDM2, MDM4, and USP7. Moreover, it appears that the effects of USP7 suppression on p53 at protein/mRNA level depend on the cell nature; however, further research is needed.
RESUMO
Background: The p53 mutation is uncommon in EpsteinBarr virus-linked gastric carcinoma, but its suppression occurs through mechanisms such as ubiquitin specific peptidase 7 (USP7) inhibitions via EpsteinBarr virus nuclear antigen-1 (EBNA1) activity. This study aimed to evaluate the effect of EBNA1 on p53-inhibiting gene expression and the impact of USP7 inhibition on p53 suppression. Methods: MKN-45 cells were transfected with the EBNA1 plasmid. A stable EBNA1 expression cell line was developed through selection based on hygromycin B resistance. Murine double minute (MDM)4, MDM2, sirtuin (SIRT)3, histone deacetylase (HDAC)1, proteasome 26S subunit, Non-ATPase (PSMD)10, USP7, and p53 expression were checked using real-time PCR. Also, cells containing EBNA1 or control plasmid were treated with GNE-6776, and the expression of the interested genes and cell survival were assessed. Results: MDM4, MDM2, and PSMD10 were significantly upregulated in the MKN-45 cell line following EBNA1 transfection. Morphological changes were observed in the cells harboring EBNA1 after 20 days. In the control cells, USP7 inhibition significantly upregulated the HDAC1, PSMD10, MDM4, and MDM2 genes after 24 h, but downregulated these genes after four days. In the EBNA1-harboring cells, MDM2, MDM4, and PSMD10 genes were significantly upregulated after 24 h, and this effect was sustained for all genes except for MDM4, even after four days. Furthermore, USP7 inhibition induced apoptosis in both cell groups. Conclusion: EBNA1 enhances the expression of p53-inhibiting genes. Two eventsp53 protein overexpression and apoptosis activationfollowed the suppression of the USP7 protein and provided evidence for its possible function. The significance of the EBNA1-USP7 interaction in p53 suppression warrants additional investigation and possibly reconsideration.