SMARCAL1 is a dual regulator of innate immune signaling and PD-L1 expression that promotes tumor immune evasion.

Genomic instability can trigger cancer-intrinsic innate immune responses that promote tumor rejection. However, cancer cells often evade these responses by overexpressing immune checkpoint regulators, such as PD-L1. Here, we identify the SNF2-family DNA translocase SMARCAL1 as a factor that favors tumor immune evasion by a dual mechanism involving both the suppression of innate immune signaling and the induction of PD-L1-mediated immune checkpoint responses. Mechanistically, SMARCAL1 limits endogenous DNA damage, thereby suppressing cGAS-STING-dependent signaling during cancer cell growth. Simultaneously, it cooperates with the AP-1 family member JUN to maintain chromatin accessibility at a PD-L1 transcriptional regulatory element, thereby promoting PD-L1 expression in cancer cells. SMARCAL1 loss hinders the ability of tumor cells to induce PD-L1 in response to genomic instability, enhances anti-tumor immune responses and sensitizes tumors to immune checkpoint blockade in a mouse melanoma model. Collectively, these studies uncover SMARCAL1 as a promising target for cancer immunotherapy.

Cytoreductive Nephrectomy Following Immune Checkpoint Inhibitor Therapy Is Safe and Facilitates Treatment-free Intervals.

Patients with metastatic renal cell cancer (mRCC) who respond to upfront immune checkpoint inhibitor (ICI) combination therapies may be treated with cytoreductive nephrectomy (CN) to remove radiographically viable primary tumors. Early data for post-ICI CN suggested that ICI therapies induce desmoplastic reactions in some patients, increasing the risk of surgical complications and perioperative mortality. We evaluated perioperative outcomes for 75 consecutive patients treated with post-ICI CN at four institutions from 2017 to 2022. Our cohort of 75 patients had minimal or no residual metastatic disease but radiographically enhancing primary tumors after ICI and were treated with CN. Intraoperative complications were identified in 3/75 patients (4%) and 90-d postoperative complications in 19/75 (25%), including two patients (3%) with high-grade (Clavien ≥III) complications. One patient was readmitted within 30 d. No patients died within 90 d after surgery. Viable tumor was present in all but one specimen. Approximately half of the patients (36/75, 48%) remained off systemic therapy at last follow-up. These data suggest that CN following ICI therapy is safe and associated with low rates of major postoperative complications in appropriately selected patients at experienced centers. Post-ICI CN may facilitate observation without additional systemic therapy in patients without significant residual metastatic disease. Patient summary: Current first-line treatment for patients with kidney cancer that has spread to other sites (metastatic cancer) is immunotherapy. For cases in which metastatic sites respond to this therapy but primary tumor is still detected in the kidney, surgical treatment of the tumor is feasible and has a low rate of complications, and may delay the need for further chemotherapy.

Mycobacterium Bovis Causing Mycotic Aneurysm Secondary to Intravesical Treatment with Bacillus Calmette-Guérin: A Case Report.

BACKGROUND: Bacillus Calmette-Guerin (BCG) is a live, attenuated strain of Mycobacterium bovis that is used in the treatment of non-muscle invasive bladder cancer (NMIBC). Vascular complications, including mycotic aneurysms, after BCG therapy are exceedingly rare. In this patient population, the diagnosis of mycotic aneurysms can be delayed or missed due to their non-specific clinical and radiologic presentation. Literature review reveals management of mycotic aneurysms attributable to BCG therapy is widely varied.2,5-8,12,15 CASE REPORT: We report a patient who presented with mycotic aneurysm formation secondary to BCG treatment for bladder cancer that was repaired with in-line reconstruction utilizing cryoartery and buttressed with omental flap. We suggest this as an alternative treatment to in-line prosthetic graft or extra-anatomic reconstruction.

Investigating factors affecting severity of large truck-involved crashes: Comparison of the SVM and random parameter logit model.

INTRODUCTION: Reducing the severity of crashes is a top priority for safety researchers due to its impact on saving human lives. Because of safety concerns posed by large trucks and the high rate of fatal large truck-involved crashes, an exploration into large truck-involved crashes could help determine factors that are influential in crash severity. The current study focuses on large truck-involved crashes to predict influencing factors on crash injury severity. METHOD: Two techniques have been utilized: Random Parameter Binary Logit (RPBL) and Support Vector Machine (SVM). Models have been developed to estimate: (1) multivehicle (MV) truck-involved crashes, in which large truck drivers are at fault, (2) MV track-involved crashes, in which large truck drivers are not at fault and (3) and single-vehicle (SV) large truck crashes. RESULTS: Fatigue and deviation to the left were found as the most important contributing factors that lead to fatal crashes when the large truck-driver is at fault. Outcomes show that there are differences among significant factors between RPBL and SVM. For instance, unsafe lane-changing was significant in all three categories in RPBL, but only SV large truck crashes in SVM. CONCLUSIONS: The outcomes showed the importance of the complementary approaches to incorporate both parametric RPBL and non-parametric SVM to identify the main contributing factors affecting the severity of large truck-involved crashes. Also, the results highlighted the importance of categorization based on the at-fault party. Practical Applications: Unrealistic schedules and expectations of trucking companies can cause excessive stress for the large truck drivers, which could leads to further neglect of their fatigue. Enacting and enforcing comprehensive regulations regarding large truck drivers' working schedules and direct and constant surveillance by authorities would significantly decrease large truck-involved crashes.

Preliminary Study on the Effect of Nitrogen Fertilisation on Phytochemical Content Quality of Gynura procumbens.

The study was carried out to examine the effects of nitrogen (N) fertilisation on phytochemical content quality, to determine types of phytochemicals compositions, and to establish phytochemicals marker compounds in Gynura procumbens. In this two factors study, three stages of harvesting (H) time and three rates of N were laid out according to Randomised Complete Block Design (RCBD). Physiological and biochemical attributes were recorded to exhibit the trend for plant quality. The results showed that, N has affected phytochemical content significantly (p < 0.05) with stronger effect on physiological and biochemical attributes (p < 0.01). The results suggested that 0 and 90 kg/ha N, respectively are highly and least effecting the Photo, Cond, TChlC, TCC, TPrc, TLiC, TPC and TFC.

Kajian ini dijalankan untuk mengkaji kesan baja nitrogen (N) terhadap kualiti kandungan fitokimia, menentukan jenis komposisi fitokimia dan menentukan sebatian penanda fitokimia dalam Gynura procumbens. Kajian ini melibatkan dua faktor; masa penuaian dan kadar N, dan mengikut RCBD. Kesan-kesan terhadap faktor fisiologi dan biokimia tumbuhan telah direkodkan dalam tiga peringkat penuaian untuk memperlihatkan corak dan kualiti pertumbuhan dan kandungan fitokimia. Nitrogen memberi kesan yang jelas terhadap kandungan fitokimia (p < 0.05), dengan kesan yang signifikan terhadap faktor fisiologi dan biokimia (p < 0.01) tumbuhan. Keputusan menunjukkan bahawa 0 dan 90 kg/ha N masing-masing memberi kesan yang sangat tinggi dan paling sedikit terhadap Photo, Cond, TChlC, TCC, TPrC, TliC, TPC dan TFC.

Blocking IL1 Beta Promotes Tumor Regression and Remodeling of the Myeloid Compartment in a Renal Cell Carcinoma Model: Multidimensional Analyses.

PURPOSE: Intratumoral immunosuppression mediated by myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages (TAM) represents a potential mechanism of immune checkpoint inhibitor (ICI) resistance in solid tumors. By promoting TAM and MDSC infiltration, IL1ß may drive adaptive and innate immune resistance in renal cell carcinoma (RCC) and in other tumor types. EXPERIMENTAL DESIGN: Using the RENCA model of RCC, we evaluated clinically relevant combinations of anti-IL1ß plus either anti-PD-1 or the multitargeted tyrosine kinase inhibitor (TKI), cabozantinib. We performed comprehensive immune profiling of established RENCA tumors via multiparameter flow cytometry, tumor cytokine profiling, and single-cell RNA sequencing (RNA-seq). Similar analyses were extended to the MC38 tumor model. RESULTS: Analyses via multiparameter flow cytometry, tumor cytokine profiling, and single-cell RNA-seq showed that anti-IL1ß reduces infiltration of polymorphonuclear MDSCs and TAMs. Combination treatment with anti-IL1ß plus anti-PD-1 or cabozantinib showed increased antitumor activity that was associated with decreases in immunosuppressive MDSCs and increases in M1-like TAMs. CONCLUSIONS: Single-cell RNA-seq analyses show that IL1ß blockade and ICI or TKI remodel the myeloid compartment through nonredundant, relatively T-cell-independent mechanisms. IL1ß is an upstream mediator of adaptive myeloid resistance and represents a potential target for kidney cancer immunotherapy.

A conserved intratumoral regulatory T cell signature identifies 4-1BB as a pan-cancer target.

Despite advancements in targeting the immune checkpoints program cell death protein 1 (PD-1), programmed death ligand 1 (PD-L1), and cytotoxic T lymphocyte-associated protein 4 (CTLA-4) for cancer immunotherapy, a large number of patients and cancer types remain unresponsive. Current immunotherapies focus on modulating an antitumor immune response by directly or indirectly expanding antitumor CD8 T cells. A complementary strategy might involve inhibition of Tregs that otherwise suppress antitumor immune responses. Here, we sought to identify functional immune molecules preferentially expressed on tumor-infiltrating Tregs. Using genome-wide RNA-Seq analysis of purified Tregs sorted from multiple human cancer types, we identified a conserved Treg immune checkpoint signature. Using immunocompetent murine tumor models, we found that antibody-mediated depletion of 4-1BB-expressing cells (4-1BB is also known as TNFRSF9 or CD137) decreased tumor growth without negatively affecting CD8 T cell function. Furthermore, we found that the immune checkpoint 4-1BB had a high selectivity for human tumor Tregs and was associated with worse survival outcomes in patients with multiple tumor types. Thus, antibody-mediated depletion of 4-1BB-expressing Tregs represents a strategy with potential activity across cancer types.

Prevalence of astrovirus, adenovirus, and sapovirus infections among Iranian children with acute gastroenteritis.

AIM: This study aimed to determine the prevalence of Human Astroviruses (HAstVs), enteric Adenoviruses (HAdVs), and Sapoviruses (SaVs) in acute diarrhea patients, as well as their relation to age, sex, and season. BACKGROUND: Acute gastroenteritis is one of the most common diseases affecting children <5 years old and viral agents with approximately >75% are the major causative agent of acute infectious diarrhea. After Rotavirus and Norovirus, the greater viral agents of acute gastroenteritis include HAstVs, HAdVs, and SaVs. To the best of our knowledge, there are sparse studies in Iran detecting at least three enteric viruses as causative agents of diarrhea simultaneously. METHODS: The sample was collected from children referring to pediatric medical centers in Tehran, Iran; they were tested for Astrovirus, enteric Adenovirus, and Sapovirus by conventional PCR method. The association of incidence of viral enteric agents was evaluated with age, sex and seasonal pattern in children <5 years old. RESULTS: The positive case number among acute gastroenteritis patients was 17/120 (14.1%). Patients ranged in age within 1-60 months, but 52.9% were aged ≤ 12 months. Males comprised the majority (70.6), and the male: female ratio was 2.4. HAstV was the most frequently detected virus (6.7%), while SaVs were detected only in 2.5% of cases. Mixed infections were not detected in these samples. The highest rate of HAstV was identified in winter (66.7%), HAdV in fall (66.7%), and SaV in winter (33.3%). CONCLUSION: These findings underscore the importance of monitoring the epidemiology of HAstV, HAdV, and SaV as causative agents of viral diarrhea infections.

Immunogenicity of prostate cancer is augmented by BET bromodomain inhibition.

BACKGROUND: Prostate cancer responds poorly to current immunotherapies. Epigenetic therapies such as BET Bromodomain inhibition can change the transcriptome of tumor cells, possibly making them more immunogenic and thus susceptible to immune targeting. METHODS: We characterized the effects of BET bromodomain inhibition using JQ1 on PD-L1 and HLA-ABC expression in two human prostate cell lines, DU145 and PC3. RNA-Seq was performed to assess changes on a genome-wide level. A cytotoxic T cell killing assay was performed in MC38-OVA cells treated with JQ1 to demonstrate increased immunogenicity. In vivo experiments in the Myc-Cap model were conducted to show the effects of JQ1 administration in concert with anti-CTLA-4 checkpoint blockade. RESULTS: Here, we show that targeting BET bromodomains using the small molecule inhibitor JQ1 decreased PD-L1 expression and mitigated tumor progression in prostate cancer models. Mechanistically, BET bromodomain inhibition increased MHC I expression and increased the immunogenicity of tumor cells. Transcriptional profiling showed that BET bromodomain inhibition regulates distinct networks of antigen processing and immune checkpoint molecules. In murine models, treatment with JQ1 was additive with anti-CTLA-4 immunotherapy, resulting in an increased CD8/Treg ratio. CONCLUSIONS: BET Bromodomain inhibition can mediate changes in expression at a genome wide level in prostate cancer cells, resulting in an increased susceptibility to CD8 T cell targeting. These data suggest that combining BET bromodomain inhibition with immune checkpoint blockade may have clinical activity in prostate cancer patients.

Using trajectory-level SHRP2 naturalistic driving data for investigating driver lane-keeping ability in fog: An association rules mining approach.

The presence of fog has a significant adverse impact on driving. Reduced visibility due to fog obscures the driving environment and greatly affects driver behavior and performance. Lane-keeping ability is a lateral driver behavior that can be very crucial in run-off-road crashes under reduced visibility conditions. A number of data mining techniques have been adopted in previous studies to examine driver behavior including lane-keeping ability. This study adopted an association rules mining method, a promising data mining technique, to investigate driver lane-keeping ability in foggy weather conditions using big trajectory-level SHRP2 Naturalistic Driving Study (NDS) datasets. A total of 124 trips in fog with their corresponding 248 trips in clear weather (i.e., 2 clear trips: 1 foggy weather trip) were considered for the study. The results indicated that affected visibility was associated with poor lane-keeping performance in several rules. Furthermore, additional factors including male drivers, a higher number of lanes, the presence of horizontal curves, etc. were found to be significant factors for having a higher proportion of poor lane-keeping performance. Moreover, drivers with more miles driven last year were found to have better lane-keeping performance. The findings of this study could help transportation practitioners to select effective countermeasures for mitigating run-off-road crashes under limited visibility conditions.

Analyzing the effect of fog weather conditions on driver lane-keeping performance using the SHRP2 naturalistic driving study data.

INTRODUCTION: Driving in foggy weather conditions has been recognized as a major safety concern for many years. Driver behavior and performance can be negatively affected by foggy weather conditions due to the low visibility in fog. A number of previous studies focused on driver performance and behavior in simulated environments. However, very few studies have examined the impact of foggy weather conditions on specific driver behavior in naturalistic settings. METHOD: This study utilized the second Strategic Highway Research Program (SHRP2) Naturalistic Driving Study (NDS) dataset to evaluate driver lane-keeping behavior in clear and foggy weather conditions. Preliminary descriptive analysis was conducted and a lane-keeping model was developed using the ordered logistic regression approach to achieve the study goals. RESULTS: This study found that individual variables such as visibility, traffic conditions, lane change, driver marital status, and geometric characteristics, as well as some interaction terms (i.e., weather and gender, surface condition and driving experience, speed limit and mileage last year) significantly affect lane-keeping ability. An important finding of this study illustrated that affected visibility caused by foggy weather conditions decreases lane-keeping ability significantly. More specifically, drivers in affected visibility conditions showed 1.37 times higher Standard Deviation of Lane Position (SDLP) in comparison with drivers who were driving in unaffected visibility conditions. CONCLUSIONS: These results provide a better understanding of driver lane-keeping behavior and driver perception of foggy weather conditions. Moreover, the results might be used to improve Lane Departure Warning (LDW) systems algorithm by allowing them to account for the effects of fog on visibility. Practical Applications: These results provide a better understanding of driver lane-keeping behavior and driver perception of foggy weather conditions. Moreover, the results might be used to improve Lane Departure Warning (LDW) systems algorithm by allowing them to account for the effects of fog on visibility.

How much does vaccination reduce the rate of HBV infection in Iranian population? a Bayesian adjustment analysis.

AIM: The aim of this research was to estimate the changing rate of odds ratio (OR) by varying degrees of hepatitis B virus (HBV) underreporting. BACKGROUND: Data registering is usually associated with extensive errors such as misclassification, under-reporting, missing data due to lack of co-operation, error prone factors, and in medical studies, inadequate diagnosis of physicians or low accuracy of laboratory tests. In the present study, which discuss the actual impact of vaccination on HBV prevention, exposure and response were prone to various errors. Furthermore, some people in the community are possibly infected to the virus while were not reported in the count of patients with HBV infection. METHODS: This was a case control study. Cases included patients with HBV referring to the gastroenterology and liver disease research center. The control group included patients without HBV who underwent a fatty liver test at Taleghani hospital laboratory. Bayesian approach and Gibbs sampling algorithm were used to estimate OR. RESULTS: According to results, misclassification rate was mild in raw data, but with an increase in degree of underreporting for 50 and 500 of unreported cases, OR increased by about half and more than double, respectively, while sensitivity diminished strikingly. CONCLUSION: Our analysis asserted that knowing the degree of underreporting is essential to accurately calculate OR and sensitivity. In addition, despite varying OR in different samples, overall the results were similar according to the pattern of exposure and response association.

Impact of organic and inorganic fertilizers on the yield and quality of silage corn intercropped with soybean.

BACKGROUND: Corn silage is an important feed for intense ruminant production, but the growth of corn relies heavily on the use of chemical fertilizers. Sustainable crop production requires careful management of all nutrient sources available on a farm, particularly in corn-based cropping systems. METHODS: Experiments were conducted to determine the appropriate technique of corn-legume intercropping in conjunction with the supplemental use of chemical fertilizers, organic manure, and biofertilizers (BFs). Acetylene reduction assays (ARAs) were also performed on corn and soybean roots. RESULTS: Combining chemical fertilizers with chicken manure (CM) in a 50:50 ratio and applying 50% NPK+50% CM+BF produced fresh forage and dry matter (DM) yields that were similar to those produced in the 100% nitrogen (N), phosphorus (P), potassium (K) treatment. Among the lone fertilizer treatments, the inorganic fertilizer (100% NPK) treatment produced the highest DM yield (13.86 t/ha) of forage and outyielded the 100% CM (9.74 t/ha) treatment. However, when CM was combined with NPK, the resulting DM yield of forage (13.86 t/ha) was the same as that resulting from 100% NPK (13.68 t/ha). Compared with CM applications alone, combinations of NPK and CM applications resulted in increased plant height, crop growth rates (CGRs) and leaf area index (LAI), but the values of these parameters were similar to those resulting from 100% NPK application. Fertilizers in which the ratio was 50% CM+50% NPK or 50% CM+50% NPK+BF resulted in protein yields that were similar to those resulting from conventional fertilizers. Similarly, the CP content did not significantly differ between applications of the 100% NPK and 50% CM+50% NPK fertilizers. The use of BFs had no significant impact on improving either the yield or quality of forage fertilized with inorganic or organic fertilizer. Lactic acid responded differently to different fertilizer applications and was significantly higher in the fertilized plots than in the unfertilized plots. Compared with treatments of lone chemical and lone organic manure fertilizers, treatments involving applications of BF and a combination of BF and NPK or CM resulted in higher ARA values. DISCUSSION: There is no simple and easy approach to increase biological nitrogen fixation (BNF) in grain legumes grown as part of a cropping system under realistic farm field conditions. Overall, evidence recorded from this study proves that, compared with corn monocrops combined with CM and chemical fertilizers, corn-soybean intercrops could increase forage yields and quality, produce higher total protein yields, and reduce the need for protein supplements and chemical fertilizers.

Formation of 6-, 8- and 10-Shogaol in Ginger through Application of Different Drying Methods: Altered Antioxidant and Antimicrobial Activity.

Gingerols and shogaols are compounds found in ginger (Zingiber officinale Roscoe); shogaols are found in lower concentration than gingerols but exhibit higher biological activities. This work studied the effects of different drying methods including open sun drying (OSD) solar tunnel drying (STD) and hot air drying (HAD) with various temperature on the formation of six main active compounds in ginger rhizomes, namely 6-, 8-, and 10-gingerols and 6-, 8-, and 10-shogaols, as well as essential oil content. Antioxidant and antimicrobial activity of dried ginger was also evaluated. High performance liquid chromatography (HPLC) analysis showed that after HAD with variable temperature (120, 150 and 180 °C), contents of 6-, 8-, and 10-gingerols decreased, while contents of 6-, 8-, and 10-shogaol increased. High formation of 6-, 8-, and 10-shogaol contents were observed in HAD (at 150 °C for 6 h) followed by STD and OSD, respectively. OSD exhibited high content of essential oil followed by STD and HAD method. Ginger-treated with HAD exhibited the highest DPPH (IC50 of 57.8 mg/g DW) and FRAP (493.8 µM of Fe(II)/g DM) activity, compared to STD and OSD method. HAD ginger exhibited potent antimicrobial activity with lower minimum inhibition concentration (MIC) value against bacteria strains followed by STD and OSD, respectively. Ginger extracts showed more potent antimicrobial activity against Gram positive bacteria than Gram negative bacteria strains. Result of this study confirmed that conversion of gingerols to shogaols was significantly affected by different drying temperature and time. HAD at 150 °C for 6 h, provides a method for enhancing shogaols content in ginger rhizomes with improving antioxidant and antimicrobial activities.

Anticancer potential of rosmarinic acid and its improved production through biotechnological interventions and functional genomics.

Rosmarinic acid (RA) is a highly valued natural phenolic compound that is very commonly found in plants of the families Lamiaceae and Boraginaceae, including Coleus blumei, Heliotropium foertherianum, Rosmarinus officinalis, Perilla frutescens, and Salvia officinalis. RA is also found in other members of higher plant families and in some fern and horned liverwort species. The biosynthesis of RA is catalyzed by the enzymes phenylalanine ammonia lyase and cytochrome P450-dependent hydroxylase using the amino acids tyrosine and phenylalanine. Chemically, RA can be produced via methods involving the esterification of 3,4-dihydroxyphenyllactic acid and caffeic acid. Some of the derivatives of RA include melitric acid, salvianolic acid, lithospermic acid, and yunnaneic acid. In plants, RA is known to have growth-promoting and defensive roles. Studies have elucidated the varied pharmacological potential of RA and its derived molecules, including anticancer, antiangiogenic, anti-inflammatory, antioxidant, and antimicrobial activities. The demand for RA is therefore, very high in the pharmaceutical industry, but this demand cannot be met by plants alone because RA content in plant organs is very low. Further, many plants that synthesize RA are under threat and near extinction owing to biodiversity loss caused by unscientific harvesting, over-collection, environmental changes, and other inherent features. Moreover, the chemical synthesis of RA is complicated and expensive. Alternative approaches using biotechnological methodologies could overcome these problems. This review provides the state of the art information on the chemistry, sources, and biosynthetic pathways of RA, as well as its anticancer properties against different cancer types. Biotechnological methods are also discussed for producing RA using plant cell, tissue, and organ cultures and hairy-root cultures using flasks and bioreactors. The recent developments and applications of the functional genomics approach and heterologous production of RA in microbes are also highlighted. This chapter will be of benefit to readers aiming to design studies on RA and its applicability as an anticancer agent.

Alpha-Mangostin-Rich Extracts from Mangosteen Pericarp: Optimization of Green Extraction Protocol and Evaluation of Biological Activity.

Since α-mangostin in mangosteen fruits was reported to be the main compound able to provide natural antioxidants, the microwave-assisted extraction process to obtain high-quality α-mangostin from mangosteen pericarp (Garcinia mangostana L.) was optimized using a central composite design and response surface methodology. The parameters examined included extraction time, microwave power, and solvent percentage. The antioxidant and antimicrobial activity of optimized and non-optimized extracts was evaluated. Ethyl acetate as a green solvent exhibited the highest concentration of α-mangostin, followed by dichloromethane, ethanol, and water. The highest α-mangostin concentration in mangosteen pericarp of 121.01 mg/g dry matter (DM) was predicted at 3.16 min, 189.20 W, and 72.40% (v/v). The verification of experimental results under these optimized conditions showed that the α-mangostin value for the mangosteen pericarp was 120.68 mg/g DM. The predicted models were successfully developed to extract α-mangostin from the mangosteen pericarp. No significant differences were observed between the predicted and the experimental α-mangostin values, indicating that the developed models are accurate. The analysis of the extracts for secondary metabolites showed that the total phenolic content (TPC) and total flavonoid content (TFC) increased significantly in the optimized extracts (OE) compared to the non-optimized extracts (NOE). Additionally, trans-ferulic acid and catechin were abundant among the compounds identified. In addition, the optimized extract of mangosteen pericarp with its higher α-mangostin and secondary metabolite concentrations exhibited higher antioxidant activities with half maximal inhibitory concentration (IC50) values of 20.64 µg/mL compared to those of the NOE (28.50 µg/mL). The OE exhibited the highest antibacterial activity, particularly against Gram-positive bacteria. In this study, the microwave-assisted extraction process of α-mangostin from mangosteen pericarp was successfully optimized, indicating the accuracy of the models developed, which will be usable in a larger-scale extraction process.

Optimization of Flavonoid Extraction from Red and Brown Rice Bran and Evaluation of the Antioxidant Properties.

Recently, the quality-by-design concept has been widely implemented in the optimization of pharmaceutical processes to improve batch-to-batch consistency. As flavonoid compounds in pigmented rice bran may provide natural antioxidants, extraction of flavonoid components from red and brown rice bran was optimized using central composite design (CCD) and response surface methodology (RSM). Among the solvents tested, ethanol was most efficient for extracting flavonoids from rice bran. The examined parameters were temperature, solvent percentage, extraction time, and solvent-to-solid ratio. The highest total flavonoid content (TFC) in red rice bran was predicted as 958.14 mg quercetin equivalents (QE)/100 g dry matter (DM) at 58.5 °C, 71.5% (v/v), 36.2 min, and 7.94 mL/g, respectively, whereas the highest TFC in brown rice bran was predicted as 782.52 mg QE/100 g DM at 56.7 °C, 74.4% (v/v), 36.9 min, and 7.18 mL/g, respectively. Verification experiment results under these optimized conditions showed that the TFC values for red and brown rice bran were 962.38 and 788.21 mg QE/100 g DM, respectively. No significant differences were observed between the predicted and experimental TFC values, indicating that the developed models are accurate. Analysis of the extracts showed that apigenin and p-coumaric acid are abundant in red and brown rice bran. Further, red rice bran with its higher flavonoid content exhibited higher nitric oxide and 2,2-diphenyl-1-picrylhydrazyl scavenging activities (EC50 values of 41.3 and 33.6 µg/mL, respectively) than brown rice bran. In this study, an extraction process for flavonoid compounds from red and brown rice bran was successfully optimized. The accuracy of the developed models indicated that the approach is applicable to larger-scale extraction processes.

Elective Nodal Irradiation Attenuates the Combinatorial Efficacy of Stereotactic Radiation Therapy and Immunotherapy.

Purpose: In the proper context, radiotherapy can promote antitumor immunity. It is unknown if elective nodal irradiation (ENI), a strategy that irradiates tumor-associated draining lymph nodes (DLN), affects adaptive immune responses and combinatorial efficacy of radiotherapy with immune checkpoint blockade (ICB).Experimental Design: We developed a preclinical model to compare stereotactic radiotherapy (Tumor RT) with or without ENI to examine immunologic differences between radiotherapy techniques that spare or irradiate the DLN.Results: Tumor RT was associated with upregulation of an intratumoral T-cell chemoattractant chemokine signature (CXCR3, CCR5-related) that resulted in robust infiltration of antigen-specific CD8+ effector T cells as well as FoxP3+ regulatory T cells (Tregs). The addition of ENI attenuated chemokine expression, restrained immune infiltration, and adversely affected survival when combined with ICB, especially with anti-CLTA4 therapy. The combination of stereotactic radiotherapy and ICB led to long-term survival in a subset of mice and was associated with favorable CD8 effector-to-Treg ratios and increased intratumoral density of antigen-specific CD8+ T cells. Although radiotherapy technique (Tumor RT vs. ENI) affected initial tumor control and survival, the ability to reject tumor upon rechallenge was partially dependent upon the mechanism of action of ICB; as radiotherapy/anti-CTLA4 was superior to radiotherapy/anti-PD-1.Conclusions: Our results highlight that irradiation of the DLN restrains adaptive immune responses through altered chemokine expression and CD8+ T-cell trafficking. These data have implications for combining radiotherapy and ICB, long-term survival, and induction of immunologic memory. Clinically, the immunomodulatory effect of the radiotherapy strategy should be considered when combining stereotactic radiotherapy with immunotherapy. Clin Cancer Res; 24(20); 5058-71. ©2018 AACR.

Phytochemical constituents, antioxidant activity, and antiproliferative properties of black, red, and brown rice bran.

BACKGROUND: In the recent years, the health benefits of the pigmented rice varieties have reported due to the presence of bioactive compounds. In this study, the phytochemical constituents (total phenolic, flavonoid and anthocyanin content) and individual phenolics and flavonoids of the extracts of sixteen genotypes of pigmented rice bran were evaluated using spectrophotometric and ultra-high performance liquid chromatography method. Antioxidative properties of the free and bound fractions were evaluated using nitric oxide and 1,1-diphenyl-2-picrylhydrazyl scavenging assays. Extracts were evaluated for antiproliferative activity against breast cancer cell lines (MCF-7 and MDA-MB-231) using the MTT assay. RESULTS: Signifficant diferences were observed in the concentrations of phytochemicals and biological activities among different pigmented rice brans. The highest phytochemical content was observed in black rice bran followed by red and brown rice bran. The concentration of free individual flavonoids and phenolic compounds were significantly higher than those of bound compounds except those of ferulic acid and p-coumaric acid. Highest antioxidant activities were observed in black rice bran, followed by red and brown rice bran extracts. Extracts of black rice bran exhibited potent antiproliferative activity, with half maximal inhibitory concentrations (IC50) of 148.6 and 119.2 mg/mL against MCF-7 and MDA-MB-231 cell lines, respectively, compared to the activity of the extracts of red rice bran (175.0 and 151.0 mg/mL, respectively) and brown rice bran (382.3 and 346.1 mg/mL, respectively). CONCLUSIONS: Black rice bran contains high levels of phytochemicals, and thus has potent pharmaceutical activity. This highlights opportunities for researcher to breed new genotypes of rice with higher nutritional values, which the food industry can use to develop new products that will compete in expanding functional food markets.

TNFα and Radioresistant Stromal Cells Are Essential for Therapeutic Efficacy of Cyclic Dinucleotide STING Agonists in Nonimmunogenic Tumors.

The cGAS-STING cytosolic DNA sensing pathway may play an integral role in the initiation of antitumor immune responses. Studies evaluating the immunogenicity of various cyclic dinucleotide (CDN) STING agonists administered by intratumoral (i.t.) injection showed potent induction of inflammation, tumor necrosis, and, in some cases, durable tumor-specific adaptive immunity. However, the specific immune mechanisms underlying these responses remain incompletely defined. The majority of these studies have focused on the effect of CDNs on immune cells but have not conclusively interrogated the role of stromal cells in the acute rejection of the CDN-injected tumor. Here, we revealed a mechanism of STING agonist-mediated tumor response that relied on both stromal and immune cells to achieve tumor regression and clearance. Using knockout and bone marrow chimeric mice, we showed that although bone marrow-derived TNFα was necessary for CDN-induced necrosis, STING signaling in radioresistant stromal cells was also essential for CDN-mediated tumor rejection. These results provide evidence for crosstalk between stromal and hematopoietic cells during CDN-mediated tumor collapse after i.t. administration. These mechanistic insights may prove critical in the clinical development of STING agonists. Cancer Immunol Res; 6(4); 422-33. ©2018 AACR.