ABSTRACT
BACKGROUND: A pro-oxidant enzyme, NADPH oxidase 4 (Nox4) has been reported to be a critical downstream effector of TGFß-induced myofibroblast transformation during fibrosis. While there are a small number of studies suggesting an oncogenic role of Nox4 derived from activated fibroblasts, direct evidence linking this pro-oxidant to the tumor-supporting CAF phenotype and the mechanisms involved are lacking, particularly in breast cancer. METHODS: We targeted Nox4 in breast patient-derived CAFs via siRNA-mediated knockdown or administration of a pharmaceutical inhibitor (GKT137831). We also determine primary tumor growth and metastasis of implanted tumor cells using a stable Nox4-/- syngeneic mouse model. Autophagic flux of CAFs was assessed using a tandem fluorescent-tagged ptfl-LC3 plasmid via confocal microscopy analysis and determination of the expression level of autophagy markers (beclin-1 and LC3B). Nox4 overexpressing CAFs depend on the Nrf2 (nuclear factor-erythroid factor 2-related factor 2) pathway for survival. We then determined the dependency of Nox4-overexpressing CAFs on the Nrf2-mediated adaptive stress response pathway for survival. Furthermore, we investigated the involvement of Birc5 on CAF phenotype (viability and collagen contraction activity) as well as the expression level of CAF markers, FAP and αSMA. CONCLUSIONS: We found that deletion of stroma Nox4 and pharmaceutically targeting its activity with GKT137831 significantly inhibited orthotopic tumor growth and metastasis of implanted E0771 and 4T1 murine mammary carcinoma cell lines in mice. More importantly, we found a significant upregulation of Nox4 expression in CAFs isolated from human breast tumors versus normal mammary fibroblasts (RMFs). Our in situ RNA hybridization analysis for Nox4 transcription on a human breast tumor microarray further support a role of this pro-oxidant in the stroma of breast carcinomas. In addition, we found that Nox4 promotes autophagy in CAFs. Moreover, we found that Nox4 promoted survival of CAFs via activation of Nrf2, a master regulator of oxidative stress response. We have further shown Birc5 is involved as a downstream modulator of Nrf2-mediated pro-survival phenotype. Together these studies indicate a role of redox signaling via the Nox4-Nrf2 pathway in tumorigenesis and metastasis of breast cancer cells by promoting autophagy and survival of CAFs.
Subject(s)
Breast Neoplasms , Cancer-Associated Fibroblasts , Animals , Breast Neoplasms/pathology , Cancer-Associated Fibroblasts/pathology , Carcinogenesis/metabolism , Cell Line, Tumor , Female , Fibroblasts/metabolism , Humans , Mice , NADPH Oxidase 4/genetics , NADPH Oxidase 4/metabolism , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , Reactive Oxygen Species/metabolism , Survivin/metabolism , Up-RegulationABSTRACT
INTRODUCTION: Lipids are a diverse group of macromolecules that occur in rice grains and are known to impact rice grain properties. Identifying the relationships between specific lipids and traits of quality is important to improve varietal selection for high quality rice. OBJECTIVES: Using untargeted lipidomics, this study aims to understand the role of lipids on different traits of quality by identifying the genotypic effect of lipids and their impact on traits of cooking and eating quality of a rice mapping population. METHODS: Lipids from milled rice grains of three sets of rice samples were screened by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) in the positive ionisation mode. Lipid features were putatively identified using analytical standards and online databases. Multivariate statistics were carried out to identify the lipid profile of varieties across three experiments. Correlation analysis was carried out between lipid features and 12 quality traits across a rice mapping population that segregates for grain physical and texture-associated traits. RESULTS: Thousands of features in rice grain lipids were detected, and were grouped into six categories-fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids and prenol lipids. A strong genotypic basis for the lipid profile was observed among the four varieties grown under five nitrogen treatments. Clear differentiation in lipid profiles between waxy and non-waxy rice was observed. Strong correlations were observed for putative lipids that form the amylose-lipid complex and with amylose content and viscosity parameters. CONCLUSIONS: This study demonstrates the strength of untargeted lipidomics in putatively determining features that differentiate varieties from each other, and reveals the role of specific lipids on the physical and textural quality of rice.
Subject(s)
Lipidomics , Lipids/analysis , Oryza/metabolism , Chromatography, High Pressure Liquid , Cooking , Eating , Humans , Mass Spectrometry , Multivariate Analysis , Quality ControlABSTRACT
BACKGROUND: Extreme heat is a public health priority in Australia with the health effects of cold poorly studied. A record-breaking cold winter prompted an institutional investigation into the epidemiology and outcomes of hypothermic presentations. AIMS: To describe the clinical and epidemiological characteristics of hypothermic emergency presentations including patient outcomes as well as gaps in practice. METHODS: This was a retrospective cohort observational study of hypothermic emergency presentations between 7 July 2009 and 1 September 2016 with a temperature of ≤35°C. Independent predictors for inpatient mortality and characteristics of exposure versus non-exposure presentations were evaluated. RESULTS: There were 217 patients with 226 presentations comprising male gender in 54%, median age 76.5 years (interquartile range (IQR) 53-88) and median initial temperature 33.3°C (IQR 31.2-34.3°C). Non-exposure presentations being found indoors, accounted for 78% overall, with elderly persons ≥65 years (P = 0.002) and multimorbidity (Charlson comorbidity index ≥4, P = 0.013) overrepresented in this subgroup. Among the non-exposure cohort, 55% were pensioners and 42% lived alone. Inpatient mortality was 11% overall and significantly higher in non-exposure versus exposure cohorts (16 vs 2%, P = 0.01). Independent predictors of inpatient mortality included heart failure (P = 0.04), metastatic malignancy (P < 0.01), chronic kidney disease (P < 0.05) and sepsis (P < 0.01). In contrast, exposure-related presentations were characterised by younger patients with intoxication due to alcohol and/or illicit drugs and psychiatric comorbidity. CONCLUSIONS: Hypothermia is a marker of clinical and socioeconomic vulnerability. The dominant presentation of the elderly patient with multimorbidity, and few social supports being found indoors, raises broader questions around the social determinants of health.
Subject(s)
Emergency Service, Hospital/statistics & numerical data , Hospital Mortality , Hypothermia/epidemiology , Aged , Aged, 80 and over , Australia/epidemiology , Comorbidity , Female , Heart Failure/epidemiology , Humans , Hypothermia/mortality , Logistic Models , Male , Middle Aged , Neoplasm Metastasis , Poverty , Renal Insufficiency, Chronic/epidemiology , Retrospective Studies , Seasons , Sepsis/epidemiology , Social Determinants of HealthABSTRACT
INTRODUCTION: Mayonnaise is an oil in water emulsion (O/W) consisting 70-80% oil. Lipid oxidation is a major cause of quality deterioration in mayonnaise. The effectiveness of antioxidants in a hetrophasic systems is highly dependent on their polarity and partitioning properties. OBJECTIVES: The aim of the present study was to determine the effect of a hydrophilic [green tea extract (GTE)] and a lipophilic [tocopherol mixture (TOC)] and BHA on lipid oxidation in mayonnaise during 60 days of storage at 38 °C and to examine the interactions between GTE and TOC, to determine possible synergistic or antagonistic effects in antioxidant activity. METHODS: The oxidative stability was studied by measuring hydroperoxides, volatile organic compounds (VOCs) and colour of mayonnaise during storage. Comprehensive analysis of VOCs was done by static headspace extraction and separation by two-dimensional gas chromatography time of flight mass spectrometry. Sensory analysis was also carried out to study the effect of storage time and antioxidant type on sensory properties of mayonnaise and to investigate the predictive ability of volatile compounds for sensory terms. RESULTS AND CONCLUSION: Addition of GTE (500 ppm) and TOC (500 ppm) increased the formation of hydroperoxides and certain VOCs. The combination of GTE with TOC improved the antioxidant efficacy compared to the individual extracts. However, sensory evaluation demonstrated that GTE promoted the development of unpleasant fishy and rancid aroma. The volatile compound methional, was significantly and positively correlated with fishy and rancid flavour. Regarding colour analysis, GTE showed the highest increase in darkening and browning during storage.
Subject(s)
Antioxidants/pharmacology , Biological Products/pharmacology , Butylated Hydroxyanisole/pharmacology , Lipids/antagonists & inhibitors , Plant Extracts/pharmacology , Plant Oils/metabolism , Tocopherols/pharmacology , Lipid Metabolism/drug effects , Plant Oils/chemistrySubject(s)
Career Choice , Humans , Victoria , Female , Male , Physicians/psychology , Specialization , Surveys and Questionnaires , Internship and Residency , AdultABSTRACT
Lipid oxidation is a primary cause of quality deterioration in mayonnaise that leads to a decrease in the nutritional and sensorial value. The evolution of volatile oxidation compounds in sunflower oil mayonnaise stored at varying temperatures for 92 days and the antioxidative effect of butylated hydroxyanisole were investigated by static headspace extraction and separation by two dimensional gas chromatography/time-of-flight mass spectrometry. Considerable differences in the headspace composition of samples stored at 4, 25 and 38 °C were found due to the different oxidation levels reached. The content of hexanal in mayonnaise at 1-5 days of storage at 38 °C could be used to predict the corresponding compound in mayonnaise at 1-62 days of storage at 25 °C. The 10 most important discriminating volatile compounds during lipid oxidation of mayonnaise (at 38 °C for 92 days) are 3-hexenal, pentanal, 2-heptenal, 2-ethylfuran, hexanal, benzeneacetaldehyde, 2-pentylfuran, 3-methylhexane, 1-pentanol and 2,4-heptadienal. More than half of these compounds have a close relationship with the initial content of linoleic acid that agrees with the fatty acid profile of sunflower oil (~ 70% linoleic acid). These volatiles could be used as additional markers of oxidation in sunflower oil mayonnaise.
ABSTRACT
KEY MESSAGE: A novel QTL cluster for chalkiness on Chr04 was identified using single environment analysis and joint mapping across 9 environments in Asia and South American. QTL NILs showed that each had a significant effect on chalk. Chalk in rice grains leads to a significant loss in the proportion of marketable grains in a harvested crop, leading to a significant financial loss to rice farmers and traders. To identify the genetic basis of chalkiness, two sets of recombinant inbred lines (RILs) derived from reciprocal crosses between Lemont and Teqing were used to find stable QTLs for chalkiness. The RILs were grown in seven locations in Asia and Latin American and in two controlled environments in phytotrons. A total of 32 (21) and 46 (22) QTLs for DEC and PGWC, most of them explaining more than 10% of phenotypic variation, were detected based on single environment analysis in T/L (L/T) population, respectively. Seven (2) and 7 (3) QTLs for DEC and PGWC were identified in the T/L (L/T) population using joined analysis across all environments, respectively. Six major QTLs clusters were found on five chromosomes: 1, 2, 4, 5 and 11. The biggest cluster at id4007289-RM252 on Chr04 was a novelty, including 16 and 4 QTLs detected by single environment analysis and joint mapping across all environments, respectively. The detected digenic epistatic QTLs explained up to 13% of phenotypic variation, suggesting that epistasis play an important role in the genetic control of chalkiness in rice. QTL NILs showed that each QTL cluster had a significant effect on chalk. These chromosomal regions could be targets for MAS, fine mapping and map-based cloning for low chalkiness breeding.
Subject(s)
Environment , Epistasis, Genetic , Oryza/genetics , Quantitative Trait Loci , Chromosome Mapping , Crosses, Genetic , Gene-Environment Interaction , Genetic Linkage , Genetic Markers , Genotype , Phenotype , Seeds , TemperatureABSTRACT
The impact of particle size of engineered nanoparticles (ENPs) on plant response has marginally been investigated under the foliar application so far. Concerning the significance of particle diameter for their properties and interaction with plants, the effect of size should be considered in the analysis of the effect of micronutrient-based ENPs on plants. It is of particular importance for ENPs containing Cu due to plants needing a relatively low amount of this element, thus there is a risk of overdosing during application as a fertilizer or pesticide. Here, we examined the biochemical and transcriptional response of barley (Hordeum vulgare L.) to Cu nanoparticles (nano-Cu) with different diameters (25 nm, 50 nm, 70 nm), microparticles (micro-Cu), and chelated Cu (EDTA-Cu). The plants suffering from Cu deficiency were foliar sprayed with Cu compounds at 1000 mg/L during the tillering stage. 1- and 7-day plants were analyzed in terms of biomass, Cu content, the activity of enzymes involved with antioxidant response, the content of low molecular weight compounds, and the expression of genes regulated metal homeostasis, aquaporins, and defense. The results showed that the Cu leaf level was differentiated over time and after 7 days it was higher under exposure to the smallest nano-Cu than other particulate Cu. Regardless of the duration of exposure, the Cu content was highest in plants treated with Cu-EDTA. The cluster analysis of all markers revealed a clear distinct response to the smallest nano-Cu and other particulate and ionic treatments. The bigger nano-Cu, depending on the markers, caused the medium effects between the nano-Cu 25 nm and micro-Cu and Cu-EDTA. The found size thresholds at the nanoscale will be useful for the fabrication of safe-by-design agrochemicals to provide crop security and attenuate environmental impact.
Subject(s)
Hordeum , Nanoparticles , Hordeum/genetics , Copper/toxicity , Copper/analysis , Edetic Acid , MineralsABSTRACT
The mechanical properties of pectin films enhanced with polyphenol-rich fruit extracts were investigated. The scavenging and reducing activity of plant extracts incorporated into the pectin films were determined using bench assays, and their antioxidant activity was correlated with a high presence of polyphenols, which were predominantly comprised of flavonoids and anthocyanins. The pectin films generated from the extracts exhibited a range of mechanical properties; tensile strength (4.99 MPa - 6.91 MPa), elongation at break (45.8 % - 52.3 %), and stiffness (1835 g mm-1 - 2765 g mm-1). To investigate the underlying relationships between plant extract composition and mechanical properties, Projection to Latent Structures (PLS) models were developed. The PLS models revealed that extracts containing high sugar and polyphenol content increase the tensile strength and moisture content of films. The elongation at break of the films was improved or diminished depending on the profile of sugar, acids, and polyphenols in the fruit extracts. Furthermore, the structures and concentration of anthocyanins and flavonoids were identified to strongly influenced the elongation at break differences. By modifying the concentration of sugars, organic acids, and polyphenols, the mechanical properties of pectin-based films can be tuned for tailored applications as food packaging materials.
Subject(s)
Plant Extracts , Polyphenols , Polyphenols/chemistry , Plant Extracts/chemistry , Pectins/chemistry , Anthocyanins , Flavonoids/analysis , Food Packaging , SugarsABSTRACT
Glycerol is a reliable solvent for extracting polyphenols from food and waste products. There has been an increase in the application of glycerol over benchmark alcoholic solvents such as ethanol and methanol for natural product generation because of its non-toxic nature and high extraction efficiency. However, plant extracts containing a high glycerol concentration are unsuitable for mass spectrometry-based investigation utilising electrospray ionization, inhibiting the ability to analyse compounds of interest. In this investigation, a solid phase extraction protocol is outlined for removing glycerol from plant extracts containing a high concentration of glycerol and their subsequent analysis of polyphenols using ultra-performance liquid chromatography coupled with quadrupole time of flight tandem mass spectrometry. Using this method, glycerol-based extracts of Queen Garnet Plum (Prunus salicina) were investigated and compared to ethanolic extracts. Anthocyanins and flavonoids in high abundance were found in both glycerol and ethanol extracts. The polyphenol metabolome of Queen Garnet Plum was 53% polyphenol glycoside derivatives and 47% polyphenols in their aglycone forms. Furthermore, 56% of the flavonoid derivates were found to be flavonoid glycosides, and 44% were flavonoid aglycones. In addition, two flavonoid glycosides not previously found in Queen Garnet Plum were putatively identified: Quercetin-3-O-xyloside and Quercetin-3-O-rhamnoside.
ABSTRACT
Fragrance in the grain is one of the most highly valued grain quality traits in rice, yet the origin and evolution of the betaine aldehyde dehydrogenase gene (BADH2) underlying this trait remains unclear. In this study, we identify eight putatively nonfunctional alleles of the BADH2 gene and show that these alleles have distinct geographic and genetic origins. Despite multiple origins of the fragrance trait, a single allele, badh2.1, is the predominant allele in virtually all fragrant rice varieties today, including the widely recognized Basmati and Jasmine types. Haplotype analysis allowed us to establish a single origin of the badh2.1 allele within the Japonica varietal group and demonstrate the introgression of this allele from Japonica to Indica. Basmati-like accessions were nearly identical to the ancestral Japonica haplotype across a 5.3-Mb region flanking BADH2 regardless of their fragrance phenotype, demonstrating a close evolutionary relationship between Basmati varieties and the Japonica gene pool. These results clarify the relationships among fragrant rice varieties and challenge the traditional assumption that the fragrance trait arose in the Indica varietal group.
Subject(s)
Betaine-Aldehyde Dehydrogenase/genetics , Evolution, Molecular , Odorants , Oryza/genetics , Plant Proteins/genetics , Alleles , Base Sequence , Betaine-Aldehyde Dehydrogenase/metabolism , Gene Frequency , Genetic Variation , Geography , Haplotypes , Molecular Sequence Data , Mutation , Oryza/classification , Oryza/enzymology , Phylogeny , Plant Proteins/metabolism , Sequence Homology, Nucleic AcidABSTRACT
OBJECTIVE: Our study assessed the reliability of a transcutaneous hemoglobin (tcHgb) measurement as compared to a standard capillary hemoglobin (cHgb) measurement in screening for iron deficiency anemia in a single university-based pediatric outpatient clinic. METHODS: Study participants included all pediatric patients requiring a hemoglobin (Hgb) assessment from July 2019 to June 2020. A tcHgb measurement was attempted on all children who received a cHgb measurement. Additional variables evaluated were age, visit type, gender, insurance type, weight, BMI percentile and presence of comorbid conditions. RESULTS: Of 777 attempts, both cHgb and tcHgb were obtained in 196 children aged 9 months to 21 years. Attempts were most successful in children > 2 years of age due to finger size and ability to remain still for one to two minutes. The mean cHgb was 12.5 ± 1.5 g/dL, mean tcHgb value 13.1 ± 2.1 g/dL, and the mean difference was 0.6 ± 2.1 g/dL (tcHgb-cHgb). An intraclass correlation coefficient was 0.29. There were no differences with regards to age, visit type, gender, insurance type, weight, BMI percentile and presence of comorbid conditions. Bland-Altman analysis displayed a lack of agreement between the tcHgb and cHgb measurements and tcHgb tended to over-estimate Hgb values when the cHgb was low. CONCLUSIONS: The measurement of a transcutaneous Hgb is less invasive for pediatric patients but has significant limitations. Smaller children (probe specifications) and movement limited the ability to obtain a tcHgb measurement in ~75% of children tested. Falsely normal tcHgb values occurred due to overestimation of hemoglobin when compared to the traditional cHgb device. The suboptimal sensitivity of the tcHgb device may cause providers to miss a diagnosis of anemia. Future research should compare both methods of Hgb assessment to the gold standard laboratory-analyzed complete blood count and use a smaller probe for children under 2, once available.
Subject(s)
Anemia, Iron-Deficiency , Hemoglobins , Adolescent , Ambulatory Care Facilities , Anemia, Iron-Deficiency/diagnosis , Child , Child, Preschool , Hemoglobins/analysis , Humans , Infant , Outpatients , Reproducibility of Results , Young AdultABSTRACT
PURPOSE: To describe the current state of telemedicine within pediatric training programs to inform development of a national telemedicine training curriculum for pediatric trainees. METHODS: We conducted an anonymous cross-sectional survey of pediatric residency (Fall 2020) and fellowship program directors (Spring 2021) on their current telemedicine practices in pediatric post-graduate training. RESULTS: Forty-eight US pediatric residency programs (n = 48/198, 24%) and 422 fellowship programs completed the survey (n = 422/872, 48%); combined response rate 44% (n = 470/1070). Pre-COVID-19, 12% (n = 57/470) of programs surveyed reported using telemedicine in their training program, but during the pandemic 71% (n = 334/470) reported telemedicine use with trainees. Over 71% (n = 334/470) agreed that a formalized curriculum is important, yet 69% (n = 262/380) of programs reporting telemedicine use either did not have a curriculum or were unsure if one existed at their program. Respondents who were unsure/not likely to add a telemedicine curriculum and/or indicated that a telemedicine curriculum would not be important (52% n = 243/470), cited "time" (55%, n = 136/243) most frequently as a barrier. CONCLUSIONS: Our needs assessment indicates marked increase in use of telemedicine with trainees by respondent pediatric training programs, with fewer than 50% reporting a formalized training curriculum and most agreeing that a curriculum is important.
Subject(s)
COVID-19 , Internship and Residency , Telemedicine , Child , Cross-Sectional Studies , Curriculum , Education, Medical, Graduate , Fellowships and Scholarships , Humans , Needs Assessment , Surveys and QuestionnairesABSTRACT
The inactivation of starch branching IIb (SBEIIb) in rice is traditionally associated with elevated apparent amylose content, increased peak gelatinization temperature, and a decreased proportion of short amylopectin branches. To elucidate further the structural and functional role of this enzyme, the phenotypic effects of down-regulating SBEIIb expression in rice endosperm were characterized by artificial microRNA (amiRNA) and hairpin RNA (hp-RNA) gene silencing. The results showed that RNA silencing of SBEIIb expression in rice grains did not affect the expression of other major isoforms of starch branching enzymes or starch synthases. Structural analyses of debranched starch showed that the doubling of apparent amylose content was not due to an increase in the relative proportion of amylose chains but instead was due to significantly elevated levels of long amylopectin and intermediate chains. Rices altered by the amiRNA technique produced a more extreme starch phenotype than those modified using the hp-RNA technique, with a greater increase in the proportion of long amylopectin and intermediate chains. The more pronounced starch structural modifications produced in the amiRNA lines led to more severe alterations in starch granule morphology and crystallinity as well as digestibility of freshly cooked grains. The potential role of attenuating SBEIIb expression in generating starch with elevated levels of resistant starch and lower glycaemic index is discussed.
Subject(s)
1,4-alpha-Glucan Branching Enzyme/genetics , Down-Regulation , Inverted Repeat Sequences , MicroRNAs/genetics , Oryza/enzymology , Plant Proteins/genetics , RNA Interference , 1,4-alpha-Glucan Branching Enzyme/chemistry , 1,4-alpha-Glucan Branching Enzyme/metabolism , Gene Expression Regulation, Plant , MicroRNAs/chemistry , MicroRNAs/metabolism , Oryza/genetics , Oryza/metabolism , Plant Proteins/chemistry , Plant Proteins/metabolism , Starch/biosynthesis , Starch/chemistryABSTRACT
A better understanding of the factors that contribute to the overall grain quality of rice (Oryza sativa) will lay the foundation for developing new breeding and selection strategies for combining high quality, with high yield. This is necessary to meet the growing global demand for high quality rice while offering producing countries additional opportunities for generating higher export revenues. Several recent developments in genetics, genomics, metabolomics and phenomics are enhancing our understanding of the pathways that determine several quality traits. New research strategies, as well as access to the draft of the rice genome, will not only advance our understanding of the molecular mechanisms that lead to quality rice but will also pave the way for efficient and targeted grain improvement.
Subject(s)
Oryza/genetics , Genomics , Metabolomics , Oryza/ultrastructure , Quantitative Trait, HeritableABSTRACT
Novel synthetic compounds, known as manganese porphyrins (MnPs), have been designed to shift the redox status of both normal cells and cancer cells. When MnPs are coupled with cancer therapies, such as radiation, they have been shown to sensitize tumor cells to treatment and protect normal tissues from damage through the modulation of the redox status of various tissue types. Until now, our preclinical studies have focused on local effects of MnPs and radiation; however, we recognize that successful outcomes for cancer patients involve control of tumor cells throughout the body. In this study, using murine orthotopic mammary tumor models, we investigated how MnPs and radiation influence the development of distant metastasis. We hypothesized that the combination of MnP (MnP/RT), such as MnTnBuOE-2-PyP5+ and radiation treatment (RT) would increase local tumor control via a shift in the intratumoral redox environment, leading to subsequent downregulation of HIF-1 in the primary tumor. Secondarily, we hypothesized that these primary tumor treatment effects would result in a reduction in pulmonary metastatic burden. Balb/c mice with orthotopic 4T1 mammary carcinomas were treated with saline, MnP, RT or MnP/RT. We found MnP/RT did extend local tumor growth delay and overall survival compared to controls and was associated with increased intratumoral oxidative stress. However, the primary tumor growth delay observed with MnP/RT was not associated with a reduced pulmonary metastatic burden. Future directions to investigate the effects of MnP/RT on the development of distant metastasis may include modifications to the radiation dose, the experimental timeline or using a murine mammary carcinoma cell line with a less aggressive metastatic behavior. Clinical trials are underway to investigate the clinical utility of MnTnBuOE-2-PyP5+ for patients undergoing radiotherapy for various tumor types. The promising preclinical data from this study, as well as others, provides support that MnP/RT has the potential to improve local tumor control for these patients.
Subject(s)
Breast Neoplasms/drug therapy , Breast Neoplasms/radiotherapy , Carcinoma/radiotherapy , Metalloporphyrins/pharmacology , Radiation Tolerance/drug effects , Animals , Breast Neoplasms/pathology , Carcinoma/drug therapy , Carcinoma/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/radiation effects , Combined Modality Therapy , Female , Humans , Manganese/pharmacology , Mice , Mice, Inbred BALB C , Oxidation-Reduction/drug effects , Oxidation-Reduction/radiation effects , Oxidative Stress/drug effects , Oxidative Stress/radiation effects , Porphyrins/pharmacologyABSTRACT
Selected Australian native fruits such as Davidson's plum, finger lime and native pepperberry have been reported to demonstrate potent antioxidant activity. However, comprehensive metabolite profiling of these fruits is limited, therefore the compounds responsible are unknown, and further, the compounds of nutritional value in these native fruits are yet to be described. In this study, untargeted and targeted metabolomics were conducted using the three fruits, together with assays to determine their antioxidant activities. The results demonstrate that targeted free and hydrolysed protein amino acids exhibited high amounts of essential amino acids. Similarly, important minerals like potassium were detected in the fruit samples. In antioxidant activity, Davidson's plum reported the highest activity in ferric reducing power (FRAP), finger lime in antioxidant capacity (ABTS), and native pepperberry in free radical scavenging (DPPH) and phosphomolybdenum assay. The compounds responsible for the antioxidant activity were tentatively identified using untargeted GC×GC-TOFMS and UHPLC-QqQ-TOF-MS/MS metabolomics. A clear discrimination into three clusters of fruits was observed using principal component analysis (PCA) and partial least squares (PLS) analysis. The correlation study identified a number of compounds that provide the antioxidant activities. GC×GC-TOFMS detected potent aroma compounds of limonene, furfural, and 1-R-α-pinene. Based on the untargeted and targeted metabolomics, and antioxidant assays, the nutritional potential of these Australian bush fruits is considerable and supports these indigenous fruits in the nutraceutical industry as well as functional ingredients for the food industry, with such outcomes benefiting Indigenous Australian communities.
ABSTRACT
Tumor associated macrophages (TAM) enhance the aggressiveness of breast cancer via promoting cancer cell growth, metastasis, and suppression of the patient's immune system. These TAMs are polarized in breast cancer with features more closely resembling the pro-tumorigenic and immunosuppressive M2 type rather than the anti-tumor and pro-inflammatory M1 type. The goal of our study was to examine primary human monocyte-derived M1 and M2 macrophages for key redox differences and determine sensitivities of these macrophages to the redox-active drug, MnTE-2-PyP5+. This compound reduced levels of M2 markers and inhibited their ability to promote cancer cell growth and suppress T cell activation. The surface levels of the T cell suppressing molecule, PD-L2, were reduced by MnTE-2-PyP5+ in a dose-dependent manner. This study also examined key differences in ROS generation and scavenging between M1 and M2 macrophages. Our results indicate that M2 macrophages have lower levels of reactive oxygen species (ROS) and lower production of extracellular hydrogen peroxide compared to the M1 macrophages. These differences are due in part to reduced expression levels of pro-oxidants, Nox2, Nox5, and the non-enzymatic members of the Nox complex, p22phox and p47phox, as well as higher levels of antioxidant enzymes, Cu/ZnSOD, Gpx1, and catalase. More importantly, we found that despite having lower ROS levels, M2 macrophages require ROS for proper polarization, as addition of hydrogen peroxide increased M2 markers. These TAM-like macrophages are also more sensitive to the ROS modulator and a pan-Nox inhibitor. Both MnTE-2-PyP5+ and DPI inhibited expression levels of M2 marker genes. We have further shown that this inhibition was partly mediated through a decrease in Stat3 activation during IL4-induced M2 polarization. Overall, this study reveals key redox differences between M1 and M2 primary human macrophages and that redox-active drugs can be used to inhibit the pro-tumor and immunosuppressive phenotype of TAM-like M2 macrophages. This study also provides rationale for combining MnTE-2-PyP5+ with immunotherapies.
Subject(s)
Macrophage Activation , Macrophages , Humans , Macrophages/metabolism , Monocytes/metabolism , Oxidation-Reduction , Reactive Oxygen Species/metabolism , STAT3 Transcription Factor/geneticsABSTRACT
The enzyme starch synthase IIa (SSIIa) in cereals has catalytic and regulatory roles during the synthesis of amylopectin that influences the functional properties of the grain. Rice endosperm SSIIa is more active in indica accessions compared to japonica lines due to functional SNP variations in the coding region of the structural gene. In this study, downregulating the expression of japonica-type SSIIa in Nipponbare endosperm resulted in either shrunken or opaque grains with an elevated proportion of A-type starch granules. Shrunken seeds had severely reduced starch content and could not be maintained in succeeding generations. In comparison, the opaque grain morphology was the result of weaker down-regulation of SSIIa which led to an elevated proportion of short-chain amylopectin (DP 6-12) and a concomitant reduction in the proportion of medium-chain amylopectin (DP 13-36). The peak gelatinization temperature of starch and the estimated glycemic score of cooked grain as measured by the starch hydrolysis index were significantly reduced. These results highlight the important role of medium-chain amylopectin in influencing the functional properties of rice grains, including its digestibility. The structural, regulatory and nutritional implications of down-regulated japonica-type SSIIa in rice endosperm are discussed.
ABSTRACT
Superoxide dismutase 3 (SOD3) is a secreted antioxidant enzyme that regulates reactive oxygen species in the microenvironment. It is also a potential tumour suppressor gene that is significantly downregulated in breast cancer. We have previously shown that its mRNA expression is inversely correlated with relapse free survival in breast cancer patients. This study aimed to investigate the correlation of SOD3 promoter DNA methylation with its expression in different molecular subtypes of breast carcinoma. We found that SOD3 expression was significantly reduced in breast carcinoma samples compared to normal tissues with the lowest levels observed in Luminal B subtype. Pyrosequencing analysis showed significant increase in methylation levels in the SOD3 promoter region (-108 and -19 from the TSS) in tumours vs normal tissues (53.6% vs 25.2%). The highest degree of correlation between methylation and SOD3 expression levels was observed in Luminal B subtype (Spearman's R = -0.540, P < 0.00093). In this subtype, the -78 CpG position is the most significantly methylated site. The Spearman's coefficient analysis also indicated the most significant correlation of DNA methylation at this site with SOD3 gene expression levels in tumours vs. normal tissues (R = -0.5816, P < 6.9E-12). Moreover, copy number variation analysis of TCGA database revealed that the more aggressive Triple Negative and Her2+ subtypes had higher levels of SOD3 gene deletion. The predominantly down-regulated expression pattern of SOD3 and the various genetic and epigenetic deregulations of its expression suggest that loss of this antioxidant promotes an advantageous tumour-promoting microenvironment in breast cancer.