Untreated Research Articles

Potential use of industrial waste-red mud in developing hybrid composites: A waste management approach

Waste reduction is one of the main aspects of cleaner production. Increasing red mud generation around the globe is creating a threat to the environment. Red mud has an attractive mineralogical composition which makes them ideal for the manufacture of composites. The current paper investigates the feasibility of utilizing red mud in the polymer matrix with coconut sheath (CS) fiber. Twelve distinct red mud composites were fabricated with silane-treated (AST), alkaline-treated (AT) and untreated (UT) CS reinforcement, as well with a differing weight percentage of red mud and investigated the mechanical properties and water and chemical resistance behaviour. AST-CS fiber composite reinforced by 20 wt% red mud displayed superior tensile, flexural and impact strength. Positive interaction of red mud with treated CS reduced the formation of voids. The chemical and water resistance test revealed the composites’ ability to withstand water and chemical attack. The effective production of polymer composites using red mud simultaneously mitigates red mud storage and pollution issues, thereby promoting a sustainable environment and an economic solution for the disposal of red mud.

Effect of a targeted treatment strategy against Ascaridia galli on egg production, egg quality and bird health in a laying hen farm

Worm control is an important aspect of the successful management of the egg production industry. Of particular concern is Ascaridia galli, which at high parasite loads affect health and production in layers. Application of a targeted treatment strategy (TT) to control A. galli has shown promise. The purpose of this study was to explore the effect of such a strategy on welfare indicators and production performance of layers. Six flocks (F1–6) on a commercial farm were allocated to three treatment groups. Flocks F1 and F4 were treated (TT) with fenbendazole at 22, 27 and 36 weeks post-placement (WPP). Flocks F2 and F5 were treated at 27 WPP (conventional treatment, CT) and hens in flocks F3 and F6 served as untreated (UT) control groups. At 19, 35 and 45 WPP twenty-five hens plus thirty eggs per flock were randomly selected. Hens were weighed and their plumage conditions (PC) were assessed. The eggs were subjected to various external and interior quality analyses. Production data such as number of eggs/hen/week, egg mass and feed conversion ratio (FCR) were calculated from raw data obtained from all flocks on a weekly basis. The number of eggs/hen/week, egg mass and FCR were higher (P < 0.05) in the TT flocks and hens had better PC both at 35 and 45 WPP compared with other flocks. No differences in body weight and physical egg quality were observed between groups except for egg shell strength which was higher (P < 0.05) in the CT flocks. These data suggest that better production performance and plumage, which suggests improved health, can be achieved through the application of a TT strategy. The insights gained from this research should help to justify the extra cost and labor associated with the TT strategy.

Impact of Peripheral α7-Nicotinic Acetylcholine Receptors on Cardioprotective Effects of Donepezil in Chronic Heart Failure Rats.

Pharmacological modulation of parasympathetic activity with donepezil, an acetylcholinesterase inhibitor, improves the long-term survival of rats with chronic heart failure (CHF) after myocardial infarction (MI). However, its mechanism is not well understood. The α7-nicotinic acetylcholine receptor (α7-nAChR) reportedly plays an important role in the cholinergic anti-inflammatory pathway. The purpose of this study was to examine whether blockade of α7-nAChR, either centrally or peripherally, affects cardioprotection by donepezil during CHF. One-week post-MI, the surviving rats were implanted with an electrocardiogram or blood pressure transmitter to monitor hemodynamics continuously. Seven days after implantation, the MI rats (n = 74) were administered donepezil in drinking water or were untreated (UT). Donepezil-treated MI rats were randomly assigned to the following four groups: peripheral infusion of saline (SPDT) or an α7-nAChR antagonist methyllycaconitine (α7PDT), and brain infusion of saline (SBDT) or the α7-nAChR antagonist (α7BDT). After the 4-week treatment, the role of α7-nAChR was evaluated using hemodynamic parameters, neurohumoral states, and histological and morphological assessment. Between the peripheral infusion groups, α7PDT (vs. SPDT) showed significantly increased heart weight and cardiac fibrosis, deteriorated hemodynamics, increased plasma neurohumoral and cytokine levels, and significantly decreased microvessel density (as assessed by anti-von Willebrand factor-positive cells). In contrast, between the brain infusion groups, α7BDT (vs. SBDT) showed no changes in either cardiac remodeling or hemodynamics. Peripheral blockade of α7-nAChR significantly attenuated the cardioprotective effects of donepezil in CHF rats, whereas central blockade did not. This suggests that peripheral activation of α7-nAChR plays an important role in cholinergic pharmacotherapy for CHF.

Formulation and Characterization of Antimicrobial Edible Films Based on Whey Protein Isolate and Tarragon Essential Oil.

The effects of heat treatment and the addition of tarragon essential oil on physical and mechanical properties of films prepared with 5% whey protein isolate (WPI) and 5% glycerol were investigated in this study. Heat treatment of the film-forming solution caused increases in thickness, moisture content, swelling degree, water vapor permeability (WVP), b*-value, ΔE*-value, transmittance values in the 200–300-nm region, transparency, and puncture resistance of the film, but decreases in water solubility, L*-value, a*-value, transmittance values in the 350–800-nm region, and puncture deformation. When incorporated with tarragon essential oil, heat-treated films have the potential to be used as antimicrobial food packaging. The addition of tarragon essential oil in film-forming solution caused increases in moisture content, solubility in water, WVP, a*-value, b*-value, ΔE*-value, and transparency of the film; decreases in transmittance values in the range of 600–800 nm; and variations in swelling degree, L*-value, transmittance values in the range of 300–550 nm, puncture resistance, and puncture deformation. Nevertheless, different tendencies were noticed in UNT (untreated) and HT (heat-treated) films with regards to transparency, light transmittance, puncture resistance, and puncture deformation. Based on these findings, HT films show improved physical and mechanical properties and, therefore, are more suitable for food-packaging applications.

Ultrasound microwave-assisted enzymatic production and characterisation of antioxidant peptides from sweet potato protein

Herein, we investigated the effects of ultrasound microwave (UM)-assisted hydrolysis using Alcalase (ALC), Flavourzyme (FLA), and their combination (ALC + FLA), on the production of sweet potato protein hydrolysates (SPPH). UM-assisted enzymatic hydrolysis significantly increased the degree of hydrolysis of SPPH compared with untreated (UN) samples. Fractions with differences in molecular weight (MW) of >10, 3–10, and < 3 kDa in SPPH from UM-assisted ALC, FLA, and ALC + FLA hydrolysis displayed higher antioxidant activities than those from UN samples. MW < 3 kDa fractions of SPPH from UM-assisted ALC and ALC + FLA hydrolysis treatments presented much stronger Fe2+-chelating activity (98.48% and 98.59%), ·OH scavenging activity (67.11% and 60.06%), and higher ORAC values (110.32 and 106.32 µg TE/mL), from which diverse peptides with potential antioxidant activities were obtained by semi-preparative HPLC and LC-MS/MS. All identified peptide sequences exhibited at least three potential antioxidant amino acids. Additionally, changes in peptide conformational structure and antioxidant amino acid composition were revealed by structure–activity relationship analysis. Thus, ultrasound microwave treatment has great potential in antioxidant peptides production.

Synthesis and characterization of polymeric films with stress-altered aluminum particle fillers

Processing and characterization of films synthesized with the blade casting method were studied. The films include 80 nm average diameter aluminum (nAl) particles combined with a solvent–binder system composed of acetone and dimethylformamide (DMF) co-solvent and Poly(vinylidene fluoride) (PVDF) binder. The nAl powder was stress-altered by annealing to 300 °C then quenching at two different cooling rates to induce elevated dilatational strain and stress within the nAl particles. Moreover, the annealing and quenching process dehydrated and dehydroxylated the surface enough to cause stress-altered particles to create more viscous slurries that produced a more porous film microstructure than untreated (UN) nAl powder. Density functional theory calculations revealed defective (i.e., dehydroxylated and dehydrated) sites on the stress-altered nAl particle surface may be responsible for the differences in nAl + PVDF reactivity that showed a higher enthalpy for aluminum-fluorine interface reactions for stress-altered nAl particles. Further, faster quenching created delamination at the core–shell interface and promoted greater energy conversion via a weaker, unsupported oxide shell that became less of a barrier for diffusion reactions. Results from this study provide insight on optimizing nAl particle properties for energy conversion using a method that could be applicable to a range of filler materials and influence bulk film properties.

Pentagalloyl glucose (PGG) partially prevents arterial mechanical changes due to elastin degradation.

Elastic fibers are composed primarily of the protein elastin and they provide reversible elasticity to the large arteries. Degradation of elastic fibers is a common histopathology in aortic aneurysms. Pentagalloyl glucose (PGG) has been shown to bind elastin and stabilize elastic fibers in some in vitro studies and in vivo models of abdominal aortic aneurysms, however its effects on native arteries are not well described. Perform detailed studies of the biomechanical effects of PGG on native arteries and the preventative capabilities of PGG for elastin degraded arteries. We treated mouse carotid arteries with PGG, elastase (ELA), and PGG+ELA and compared the wall structure, solid mechanics, and fluid transport properties to untreated (UNT) arteries. We found that PGG alone decreased compliance compared to UNT arteries, but did not affect any other structural or biomechanical measures. Mild (30 sec) ELA treatment caused collapse and fragmentation of the elastic lamellae, plastic deformation, decreased compliance, increased modulus, and increased hydraulic conductance of the arterial wall compared to UNT. PGG+ELA treatment partially protected from all of these changes, in particular the plastic deformation. PGG mechanical protection varied considerably across PGG+ELA samples and appeared to correlate with the structural changes. Our results provide important considerations for the effects of PGG on native arteries and a baseline for further biomechanical studies on preventative elastic fiber stabilization.

Gene Expression Profile and Acute Gene Expression Response to Sclerostin Inhibition in Osteogenesis Imperfecta Bone.

ABSTRACTSclerostin antibody (SclAb) therapy has been suggested as a novel therapeutic approach toward addressing the fragility phenotypic of osteogenesis imperfecta (OI). Observations of cellular and transcriptional responses to SclAb in OI have been limited to mouse models of the disorder, leaving a paucity of data on the human OI osteoblastic cellular response to the treatment. Here, we explore factors associated with response to SclAb therapy in vitro and in a novel xenograft model using OI bone tissue derived from pediatric patients. Bone isolates (approximately 2 mm3) from OI patients (OI type III, type III/IV, and type IV, n = 7; non‐OI control, n = 5) were collected to media, randomly assigned to an untreated (UN), low‐dose SclAb (TRL, 2.5 μg/mL), or high‐dose SclAb (TRH, 25 μg/mL) group, and maintained in vitro at 37°C. Treatment occurred on days 2 and 4 and was removed on day 5 for TaqMan qPCR analysis of genes related to the Wnt pathway. A subset of bone was implanted s.c. into an athymic mouse, representing our xenograft model, and treated (25 mg/kg s.c. 2×/week for 2/4 weeks). Implanted OI bone was evaluated using μCT and histomorphometry. Expression of Wnt/Wnt‐related targets varied among untreated OI bone isolates. When treated with SclAb, OI bone showed an upregulation in osteoblast and osteoblast progenitor markers, which was heterogeneous across tissue. Interestingly, the greatest magnitude of response generally corresponded to samples with low untreated expression of progenitor markers. Conversely, samples with high untreated expression of these markers showed a lower response to treatment. in vivo implanted OI bone showed a bone‐forming response to SclAb via μCT, which was corroborated by histomorphometry. SclAb induced downstream Wnt targets WISP1 and TWIST1, and elicited a compensatory response in Wnt inhibitors SOST and DKK1 in OI bone with the greatest magnitude from OI cortical bone. Understanding patients' genetic, cellular, and morphological bone phenotypes may play an important role in predicting treatment response. This information may aid in clinical decision‐making for pharmacological interventions designed to address fragility in OI. © 2020 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

2084-P: Hepatic Fat Accumulation Underlies the Inability of Glucokinase Activator to Achieve a Sustained Hypoglycemic Effect in db/db Mice

Background: Previous clinical trials have demonstrated that glucokinase activators (GKAs) do not achieve a sustained hypoglycemic effect; however, the mechanism for this has not been determined. Aims: We aimed to clarify the mechanisms for the loss of GKA hypoglycemic effect. Method: Six-week-old db/db mice were fed standard chow and divided into three groups as follows: Untreated (UT), 0.04% GKA (G), or 0.005% ipragliflozin, a sodium-glucose cotransporter 2 inhibitor that has a long-term hypoglycemic effect in db/db mice (S). After measuring body weight (BW) and blood glucose (BG) for 28 days, pancreatic beta-cell mass and hepatic triglyceride (TG) content were analyzed. Results: The three groups did not differ in BW. A hypoglycemic effect persisted until Day 28 in S; however, in G it lasted only 6 days (Day 6: UT 454 ± 84 mg/dL, G 257 ± 85 mg/dL, S 224 ± 55 mg/dL, Day 28: UT 588 ± 102 mg/dL, G 508 ± 129 mg/dL, S 217 ± 30 mg/dL). Insulin sensitivity on insulin tolerance testing did not differ between Days 9 and 23. Glucose tolerance was improved in G and S on Days 4 and 11; however, this improvement had disappeared by Day 25. Only S exhibited increased fasting insulin-to-BG ratio on Days 11 and 25. On Day 28, pancreatic beta-cell mass and proliferation were increased in S, but not in G. Although the three groups did not differ in visceral or subcutaneous fat weight, liver weight was increased in G on Days 6 and 28. Hepatic TG content was increased in G on Day 6 (UT 130 ± 18 mg/g liver, G 226 ± 20 mg/g liver, S 39 ± 7 mg/g liver), and mRNA expression related to lipogenesis, including Fas, Elovl6 and Scd1, was upregulated in G. Conclusion: Our results indicate that GKA does not increase insulin secretion or pancreatic beta-cell mass in db/db mice. However, GKA does increase hepatic TG in db/db mice, suggesting that GKA increases glucose utilization in the liver. However, it might also promote lipogenesis and hepatic fat accumulation, leading to its hypoglycemic effect being unsustained. Disclosure S. Kawata: None. A. Nakamura: None. H. Miyoshi: Research Support; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Novo Nordisk Inc., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd. Speaker’s Bureau; Self; Astellas Pharma Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Kowa Company, Ltd., MDS CO. LTD., Mitsubishi Tanabe Pharma Corporation, Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Sanofi K.K., Sumitomo Dainippon Pharma Co., Ltd. Other Relationship; Self; Abbott, Boehringer Ingelheim Pharmaceuticals, Inc., Kowa Company, Ltd., Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd. K. Tsuchida: None. K. Omori: None. K. Takahashi: None. H. Nomoto: None. H. Kameda: None. K. Cho: None. Y. Terauchi: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Novo Nordisk A/S, Sanofi. Consultant; Self; Astellas Pharma Inc. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Merck Sharp &amp; Dohme Corp., Novartis Pharmaceuticals Corporation, Novo Nordisk A/S, Ono Pharmaceutical Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Takeda Pharmaceutical Company Limited. Speaker’s Bureau; Self; Astellas Pharma Inc., AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Kowa Company, Ltd., Medscape, Medtronic, Merck Sharp &amp; Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Novo Nordisk A/S, Ono Pharmaceutical Co., Ltd., Sanofi, Sanwa Kagaku Kenkyusho, Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Terumo Medical Corporation. T. Atsumi: Consultant; Self; Chugai Pharmaceutical Co., Ltd., Eli Lilly Japan K.K., GlaxoSmithKline plc., Pfizer Inc., UCB Japan Co. Ltd. Speaker’s Bureau; Self; AbbVie Inc., Alexion Pharmaceuticals, Inc., Astellas Pharma Inc., Bristol-Myers Squibb, Chugai Pharmaceutical Co., Ltd., Daiichi Sankyo, Eisai Co., Ltd., Eli Lilly Japan K.K., Gilead Sciences, Inc., Mitsubishi Tanabe Pharma Corporation, Otsuka Pharmaceutical Co., Ltd., Pfizer Inc., Takeda Pharmaceutical Company Limited, UCB Japan Co. Ltd.

Stress-altered aluminum powder dust combustion

Aluminum powder was thermally stressed by annealing and quenching, then the powder was non-uniformly dispersed in air and examined for dust combustion behavior as a function of stress-altering conditions. An explosion chamber with a powder injector, spark gap igniter, pressure sensor, spectrometer, and high-speed camera was used for experimentation. Aluminum powder was annealed to 573 K, held for 15 min, and quenched at a rate of 200 K/min (pre-stressed, PS) or 900 K/min (super-quenched, SQ). The untreated (UN), PS, and SQ Al powders were injected into the chamber, and pressure, temperature, and flame spreading behavior were analysed. SQ Al powder exhibited lower pressurization rates than that of PS Al, which was also lower than that of UN Al. Surface modifications to the stress-altered powders may affect their dispersion and suspension in the air environment, which affects flame spreading and pressurization rate. Specifically, annealing powders caused the removal of surface hydration that had two effects: increased the surface energy of the particles (confirmed with density functional theory calculations) and decreased surface roughness (suggested from previous work revealing loss of a nanostructure at the surface with annealing). These two surface modifications may inhibit powder dispersion such that pressurization rate is reduced compared with UN Al powder.

Lipase Treatment of Dietary Krill Oil, but Not Fish Oil, Enables Enrichment of Brain Eicosapentaenoic Acid and Docosahexaenoic Acid.

Currently available omega-3 fatty acid supplements do not enrich the docosahexaenoic acid (DHA) of the adult brain because they are absorbed as triacylglycerol, whereas the transporter at the blood brain barrier requires lysophosphatidylcholine (LPC)-DHA. The hypothesis that treatment of krill oil (KO), which contains DHA/eicosapentaenoic acid (EPA) at the SN2 position of phosphatidylcholine, with SN1-specific lipase will generate LPC-DHA/EPA and which can be absorbed intact and transported into the brain, is tested. KO and fish oil (FO) are treated with Mucor meihei lipase, incorporated into AIN 93G diet, and fed to 2-month-old mice for 30 days. Fatty acid composition is analyzed by gas chromatography/mass spectroscopy. Brain derived neurotrophic factor (BDNF) is measured by ELISA. Lipase-treated (LT) KO increases brain DHA and EPA, respectively, 5-and 70-fold better than untreated (UT) KO. FO, whether lipase-treated or not, has no effect on brain DHA/EPA. LTKO is also more efficient in enriching liver DHA/EPA, but less efficient than UTKO and FO in enriching adipose tissue and heart. Brain BDNF is significantly increased by LTKO, but only marginally by other preparations. Pretreatment of dietary KO with lipase enables it to efficiently increase brain DHA/EPA because of the generation of LPC-DHA/EPA.

Role of nitric oxide and reactive oxygen species in static magnetic field pre-treatment induced tolerance to ambient UV-B stress in soybean.

The experiments were conducted for the estimation of the mitigating effect of the static magnetic field (SMFof 200 mT for 1 h) treatment on soybean under ambient UV-B stress. The SMF treated (MT) and untreated (UT) seedswere grown inside iron cages covered with polyester filters for the purpose to filter UV-A + B (< 400nm) and UV-B (< 300nm) radiations, polythene filter control (FC) transparent for UV (280-400nm), and open controls (OC) were without any filters. Our results indicated that specific leaf weight, efficiency of PS II, activity of carbonic anhydrase (CA) and nitrogenase (NRA), nucleic acid and protein content, nitric oxide (NO) and yield were significantly decreased in plants of untreated seeds under UV-B stress. SMF treatment to the soybean seeds was observed to mitigate the adverse effect of ambient UV-B with a significant enhancement in above-measured parameters in plants when compared with plants of untreated seeds grown underOC/FC conditions. Chlorophyll a fluorescence transition curve (OJIP-curve) from SMF treated and UV excluded plants has shown a higher fluorescence yield especially for I-P phase as compared to the plants grown in ambient UV-B stress. Reduction in the level of superoxide anion radicle ( ), hydrogen peroxide (H2O2), malondialdehyde (MDA) and proline content with a remarkable increase in DNA, RNA, protein and NO content, increased photosynthetic efficiency and nitrogen fixation in the leaves of soybean suggested the ameliorating effect of SMF pre-treatment against ambient UV-B induced damage. Consequently, SMF-pretreatment increased the tolerance of soybean seedlings to ambient UV-B stress as compared to the untreated seeds. The increase in carbon and nitrogen fixation ability due to SMF pre-treatment and the omission of solar UV radiation impact can be a direction for the purpose to improve the crop yield. Evaluation of the consequences of SMF treated seeds under ambient UV-B stress, and the plants from untreated seeds under solar UV exclusion indicated parallelism among the two effects.

Performance Evaluation of Cryogenic Treated and Untreated Carbide Inserts during Machining of AISI 304 Steel

The cutting tool in the machining process plays an important role as it acts on the working material. There are a few methodologies have been persued to improve tool life, for example traditional cooling, single layer coating, multilayer coating, heat treatment process, nitrogen cooling and latest being the cryogenic treatment which reported a significant improvement in cutting tool life, chip morphology, reduction in heat generation. Hence, the cryogenic treatment is emerged as the sustainable machining process. This paper presents machining of AISI 304 steel using both cryogenic treated (CT) and untreated (UT) cutting tool insert. The commercially available uncoated carbide insert has been cryogenically treated at -196°C for 24 hours soaking period. The machining test has been conducted under four different cutting speeds. The material characterization of cutting insert is studied by using scanning electron microscopy (SEM), hardness test, and microscopic image analysis has been carried out before and after cryogenic treatment. The cutting tool performance is assessed in terms of of wear, cutting temperature, chip morphology, surface roughness under the influence of cryogenic machining and the results are contrast with UT one. The exploratory findings reveals that the deep cryogenic treatment (DCT) with 24 hours soaking period, performed better wear resistance and improved surface roughness of the cutting tool. Also considerable reduction in the flank wear, crater wear, cutting temperature is obtained and found improved chip morphology.

Abstract A101: Single-cell analysis reveals the pivotal role of the innate immune compartment in aCTLA-4 antitumor response

Abstract Immunotherapies have revolutionized and continue to revolutionize cancer patient care; however, much of our understanding of the mechanism of action of these therapies is currently limited. The accumulation of regulatory T cells (Tregs) in the tumor hampers antitumor activity and correlates with bad prognosis in several human cancers. Anti-CTLA-4 monoclonal antibodies (mAbs) have been extensively studied, and their activity depends both on the blockade of the CTLA-4 coinhibitory molecule, as well as the intratumoral depletion of Tregs, increasing effector cell abundance, and favoring tumor rejection. Fc-gamma receptor (FcgR) coengagement has been proven to be important for the action of aCTLA-4, in addition to its Treg-depleting activity via antibody-dependent cellular cytotoxicity (ADCC); however, it is unclear what are the cellular changes involved. We therefore wished to dissect the differences between aCTLA-4 mIgG1, an antibody that only blocks the CTLA-4 receptor, and aCTLA-4 mIgG2a, which has a dual activity of both blocking the receptor and depleting Tregs. To address this, we performed single-cell RNA-seq of infiltrating leukocytes from tumors in mice treated with aCTLA-4 mIgG1, mIgG2a, or left untreated (UT). This high-resolution comparison revealed unique cellular profiles generated by each treatment. While the blocking-only mIgG1 was similar to UT, mIgG2a with ADCC effector function demonstrated major changes. Tumors in mIgG2a treated mice showed an immediate decline of immune suppressive macrophages, and the emergence of proinflammatory monocytes, as well as NK cells and naive CD8 T cells. In addition, mIgG2a-treated mice displayed a gradual increase of CD4 T-cell subsets throughout treatment. Finally, a vast tissue repair signature was observed in later time points of aCTLA-4 mIgG2a treatment, comprising MRC1+ macrophages, neutrophils, and myeloid-derived suppressor cells (MDSCs), while CD8 T-cell abundance declined. In summary, our findings provide an in-depth view of the differences in mechanisms of action between an aCTLA-4 blocking mAb and an optimized blocking-depleting mAb, underscoring how FcgRs coengagement leads to enhanced antitumor response via the innate immune compartment. Citation Format: Ido Yofe, Adam Jelinski, Isabelle Solomon, Tomer Landsberger, Marc Robert de Massy, Karl Peggs, Sergio Quezada, Ido Amit. Single-cell analysis reveals the pivotal role of the innate immune compartment in aCTLA-4 antitumor response [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr A101.

Energy grass/polylactic acid composites and pretreatments for additive manufacturing

The aim of this study was to find the effect of ball-milling pretreatment of silvergrass and comparing the effects of pretreatment on common lignocellulosic materials such as reed, switchgrass and pennisetum to evaluate the potential of raw materials for additive manufacturing. The size of ball-milled material was much smaller than the nozzle diameter of the 3D printer and the melting point of composite was lower than that of pure PLA according to differential scanning calorimetry. The structure of the surface became smoother distinctly and biomass dispersed better in PLA matrix after ball-milling based on scanning electron microscopy. X-ray diffraction results showed that the ball-milling treatment influenced the performance of the composite by destroying the crystalline region and increasing the amorphous region of raw materials. Besides, the PLA/silvergrass composites possessed better mechanical properties of tensile strength, Young’s modulus and impact strength of 23.1 MPa, 652.9 MPa and 38.67 kJ/m2, respectively. Compared to the composites that were prepared by alkali-H2O2 (AH) and by steam explosion (SE) treatment, the composite that was prepared by acid treatment (AT) exhibited higher tensile strength, Young’s modulus and impact strength of 26.94 MPa, 720.36 MPa and 40 kJ/m2 respectively. Silvergrass/PLA composite shows better thermal stability compared to pure PLA for additive manufacturing. Moreover, AT works as a promising pretreatment method in promoting the properties of wood-plastic composite compared to UT (untreated), AH and SE. The silvergrass/PLA composite could be used for additive manufacturing after ball-milling combined with AT.

Evaluation of the Biofield Energy Treated Novel Test Formulation on Overall Organs Health Specific Biomarkers

The aim was to study the impact of the Biofield Energy Treated test formulation on the function of vital organs viz. bones, heart, liver, lungs, and brain in various cell-based assays. The test formulation and the cell media were divided into two parts; one part was untreated (UT) and other part received the Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, John Suzuki, USA and was labeled as the Biofield Energy Treated (BT) test formulation/media. Cell viability data suggested that the test formulation was safe and non-toxic in nature in six different cells. The Biofield Treated medium (BT-Med) + Biofield Treated Test Item (BT-TI) group showed 91.6%, 56.9%, and 114.5% restoration of cell viability at 1, 10, and 25 µg/mL, respectively in human cardiac fibroblasts cells (HCF) compared to the UT-Med + UT-TI group. Moreover, BT-Med + BT-TI group showed 70.6%, 126.3%, and 60.2% restoration of cell viability at 1, 25, and 63 µg/mL, respectively; while 78.5% in the UT-Med + BT-TI group in human hepatoma cells (HepG2) compared to untreated. Furthermore, 72.6% (at 0.1 µg/mL), 57.4% (at 25 µg/mL), and 90.4% (at 63 µg/mL) restoration of cell viability was observed in adenocarcinomic human alveolar basal epithelial cells (A549) by BT-Med + UT-TI, UT-Med + BT-TI, and BT-Med + BT-TI groups, respectively compared to the untreated. The alkaline phosphatase (ALP) level was significantly increased by 82.2% and 106.6% in the UT-Med + BT-TI and BT-Med + UT-TI groups, respectively at 10 µg/mL; while 80.2% (at 10 µg/mL) in the BT-Med + BT-TI group in human bone osteosarcoma cells (MG-63) compared to the untreated. Additionally, the level of ALP was significantly increased by 90.2% (at 0.1 µg/mL) and 78.3% (at 10µg/mL) in the BT-Med + BT-TI group in human endometrial adenocarcinoma cells (Ishikawa) compared to the untreated. The percent protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 63.7%, 59.6%, and 63.3% at 1, 10, and 25 µg/mL, respectively in the BT-Med + BT-TI group; while 122.2% (at 0.1 µg/mL) in the UT-Med + BT-TI group as compared to the untreated in HCF cells. The percent protection of HepG2 (liver) cells (decreased of ALT activity) was significantly increased by 62.8% and 153.2% at 1 µg/mL in the UT-Med + BT-TI and BT-Med + BT-TI groups, respectively compared to untreated in HepG2 cells. The percent protection of A549 (lungs) cells (increased of SOD activity) was significantly increased by 133.1%, 153.8%, and 107.5% in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively at 10 µg/mL compared to untreated. Serotonin level was significantly increased by 75.7% (at 10 µg/mL), 124.1% (at 10 µg/mL), and 73.5% (at 10 µg/mL) in the BT-Med + UT-TI group at 10, 25, and 63 µg/mL, respectively; while 107.7% (at 25 µg/mL) in the BT-Med + BT-TI group as compared to the untreated in human neuroblastoma cells (SH-SY5Y). The relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 162.2% (at 10 µg/mL), 193.3% (at 1 µg/mL), and 148.7% (at 0.01 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to the untreated in MG-63 cells. Overall, these results suggest that Biofield Treated test formulation significantly improved the bones, heart, liver, lungs, and brain functional enzyme biomarkers. Altogether data suggest that the Biofield Energy Treatment (The Trivedi Effect®) can be useful to protect and maintain the normal function of each vital organ such as lungs, liver, heart, brain, and bones. Therefore, The Trivedi Effect® can be used as a complementary and alternative therapy against several disorders such as coronary artery disease, heart attack, heart failure, arrhythmias, congenital heart disease, cirrhosis, cardiomyopathy, Wilson disease, liver cancer, hemochromatosis, pneumonia, asthma, cystic fibrosis, emphysema, chronic bronchitis, osteoporosis, etc.

In Vitro Cell-Based Biomarkers Study of Vital Organs: Impact of the Biofield Energy Based Test Formulation

The present study was undertaken to evaluate the impact of Biofield Energy Treated test formulation using multiple cell-lines. The test formulation and cell media (Med) was divided into two parts; one part was untreated (UT) and other part received Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Krista Joanne Callas, USA and labeled as Biofield Energy Treated (BT) test item (TI)/Med. Based on cell viability, test formulation was found safe. Cytoprotective action of test formulation showed significant restoration of cell viability by 89.9% and 106.4% in human cardiac fibroblasts cells (HCF) cells, while improved restoration of cell viability by 77.3% and 69% in HepG2 cells compared to untreated. Cellular restoration in A549 cells was also improved by 141.2% and 157.1% compared to untreated. ALP activity was significantly increased by 118.7% and 140.7% in UT-Med + BT-TI and BT-Med + UT-TI, respectively at 0.1 µg/mL than untreated. Percent cellular protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 89.9% and 106.4% in UT-Med + BT-TI and BT-Med + BT-TI, respectively than untreated. HepG2 cells protection (decreased ALT activity) was increased by 59.8% in BT-Med + BT-TI than untreated. Superoxide dismutase (SOD) level was increased by 22.8% in BT-Med + BT-TI than untreated. Serotonin level was significantly increased by 361.7% and 197.6% in BT-Med + UT-TI and BT-Med + BT-TI, respectively than untreated in human neuroblastoma cells (SH-SY5Y). However, relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 116.5%, 214.7%, and 241.5% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively than untreated in MG-63 cells. Overall, data showed a significant improvement of organ-specific functional enzyme biomarkers. Thus, Biofield Energy Treated Test formulation (the Trivedi Effect®) would be useful for multiple organs health that can be beneficial against coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

Impact of Biofield Energy Treatment Based Test Formulation on Vital Organ Health Specific Biomarkers Using Cell Line Study

Multiple organ dysfunction syndrome or failure is one of the major concerns against healthcare services in order to maintain the normal function. The present study aimed to explore the impact of the Biofield Energy Treated test formulation on the function of vital organs such as bones, heart, liver, lungs, and brain using standard activity parameters in specific cell-based assays. The test formulation and cells medium was divided into two parts, one untreated (UT) and other part received the Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Ariadne Esmene Afaganis, Canada and was labeled as the Biofield Treated (BT) test formulation/media. The test formulation was tested for cell viability, and the data suggested that the test formulation was found safe and non-toxic against all the cell lines. Cytoprotective activity among the experimental groups showed a significant improved activity by 94.4% at 1 µg/mL in untreated medium (UT-Med) + Biofield Treated Test Item (BT-TI) group in human cardiac fibroblasts cells (HCF) cells, while 84.4% at 10 µg/mL in BT-Med + BT-TI groups in human hepatoma cells (HepG2), and 124% increased cytoprotective action at 1 µg/mL in UT-Med + BT-TI group in adenocarcinomic human alveolar basal epithelial cells (A549) cells as compared with the untreated test group. ALP activity in MG-63 cells was significantly increased by 85.9% at 10 µg/mL in the UT-Med + BT-TI group, while in Ishikawa cells showed maximum increased ALP activity by 59.2% at 0.1 µg/mL in BT-Med + BT-TI groups as compared to the untreated group. The percent protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 53% and 40.5% at 1 and 10 µg/mL concentrations respectively, in UT-Med + BT-TI group, while BT-Med + UT-TI group showed increased protection by 68.5%, 70.7%, and 16.8% at 0.1, 1, and 10 µg/mL respectively, and 86.5%, 62.5%, and 34.2% improved cellular protection at 0.1, 1, and 10 µg/mL respectively, in BT-Med + BT-TI group as compared to the untreated test group. The percent protection of HepG2 (liver) cells (decreased of ALT activity) was reported by 33.5%, 63.2%, and 99.2% at 10 µg/mL in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to the untreated group. Cellular protection of A549 (lungs) cells (increased of SOD activity) in terms of percentage was increased by increased by 39.8% (at 10 µg/mL), 44% (at 25.5 µg/mL), and 59.7% (at 25.5 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to untreated group. Serotonin level was significantly increased by 59.2% (at 0.1 µg/mL), 190.3% (at 0.1 µg/mL), and 201% (at 1 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively compared to untreated in human neuroblastoma cells (SH-SY5Y). However, the relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 159.1% (at 50 µg/mL), 212.7% (at 1 µg/mL), and 278.3% (at 10 µg/mL) in the UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI groups, respectively as compared to the untreated in MG-63 cells. Thus, the present data concluded that the overall multiple organ health using various standard biomarkers in specific cell lines were significantly improved with respect to health of bones, heart, liver, lungs, and brain after treatment with the Biofield Energy treated test formulation (The Trivedi Effect®). Thus, it can be used as a complementary and alternative therapy approach against many multiple organ disorders such as coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

Cell-Based Vital Organs Specific Biomarkers Assessment using Biofield Energy Based Novel Test Formulation

The aim of the present study was to determine the impact of Biofield Energy Treated test formulation using six differentcell-lines. The test formulation/item (TI) and cell media (Med) was divided into two parts; one part was untreated (UT) and other part received Biofield Energy Treatment remotely by a renowned Biofield Energy Healer, Janice Patricia Kinney, USA and labeled as Biofield Energy Treated (BT) test item (TI)/media. Based on cell viability assay, test formulation was found as safe at tested concentrations. Cytoprotective activity of test formulation showed a significant restoration of cell viability by 60.6% (10 µg/mL), 67.5% (63.75 µg/mL), and 117.5% (63.75 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, BT-Med + BT-TI, respectively compared to untreated in human cardiac fibroblasts cells (HCF) cells. Moreover, restoration of cell viability was improved by 64% and 127.3% in UT-Med + BT-TI and BT-Med + UT-TI, respectively at 1 µg/mL compared to untreated in human liver cancer (HepG2) cells. Cellular restoration in A549 cells was improved by 314% and 112.3% at 1 µg/mL in BT-Med + UT-TI and BT-Med + BT-TI, respectively than untreated. ALP activity in Ishikawa cells was significantly increased by 175.5%, 547.2%, and 220.8% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively at 0.1 µg/mL as compared to untreated. Additionally, in MG-63 cells showed increased ALP activity by 76.9%, 78.4%, and 79% in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively at 50 µg/mL compared to untreated. The percent cellular protection of HCF (heart) cells (decreased of LDH activity) was significantly increased by 60.6% (10 µg/mL), 67.5% (63.75 µg/mL), and 117.5% (63.75 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively as compared to untreated. An improved HepG2 cells protection (represents decreased ALT activity) by 115.1% (1 µg/mL), 42.5% (25.5 µg/mL), and 60.8% (10 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, BT-Med + BT-TI, respectively as compared to untreated. Percentage cellular protection of A549 (lungs) cells (represents increased of SOD activity) was significantly increased by 191.1% and 81.4% at 0.1 µg/mL in UT-Med + BT-TI and BT-Med + BT-TI, respectively as compared to untreated. Serotonin level was significantly increased by 31.8% (10 µg/mL) and 56.9% (25.5 µg/mL) in UT-Med + BT-TI and BT-Med + BT-TI, respectively compared to untreated in human neuroblastoma cells (SH-SY5Y). Relative quantification (RQ) of vitamin D receptor (VDR) was significantly increased by 304.3% (0.01 µg/mL), 128.4% (0.1 µg/mL), and 240% (0.1 µg/mL) in UT-Med + BT-TI, BT-Med + UT-TI, and BT-Med + BT-TI, respectively compared to untreated in MG-63 cells. Thus, Biofield Energy Treated test formulation (The Trivedi Effect®) significantly improved organ specific functional biomarkers and would be useful for multiple organs health related to coronary artery disease, arrhythmias, congenital heart disease, cardiomyopathy, cirrhosis, liver cancer, hemochromatosis, asthma, chronic bronchitis, cystic fibrosis, osteoporosis, etc.

Expression and Role of Nucleotide-Binding Oligomerization Domain 2 (NOD2) in the Ocular Surface of Murine Dry Eye.

To investigate expression and role of nucleotide-binding oligomerization domain 2 (NOD2) in the ocular surface of experimental dry eye (EDE), which is a nod-like receptor member and is involved in innate immune response. C57/BL6 female mice were divided into the groups: untreated (UT), EDE, and NOD2 knockout (KO) mice exposed to desiccating stress for 14 days. Clinical parameters and levels of inflammatory cytokine were measured at 3,5,7, and 14 days. Immunofluorescent staining for NOD2 and Western blot for RIP2 and NF-κB were performed at 14 days. Flow cytometry, PAS staining and TUNEL staining were performed. After EDE induction, NOD2 was expressed in the corneal epithelium of the EDE group. The EDE group showed a significantly increased RIP2 expression compared to the UT and NOD2-KO groups. A significantly lower expression of NF-κB and lower levels of IL-1β, IL-6, IFN-γ, and TNF-α were noted in the NOD2-KO group than in the EDE group. The NOD2-KO group had lower CD11b+ and CD4+CCR5+ T cells, TUNEL-positive cells and corneal staining score and higher density of conjunctival goblet cell density, tear volume, and tear film break-up time than the EDE group. The UT group showed significant differences in inflammatory and clinical parameters compared to the EDE and NOD2-KO groups. The NOD2 receptor pathway induced inflammation and apoptosis by activation of RIP2 and NF-κB on the ocular surface of EDE, thereby reducing tear secretion. Therefore, NOD2 pathway may be involved in the pathogenesis of dry eye.