TC Treatment Research Articles

Taraxacum coreanum (Korean Dandelion) Extract Protects against Lipopolysaccharide-Induced Blood-Brain Barrier Destruction via Regulation of Tight Junctions and Inflammatory Responses in bEnd.3 Cells.

Dysfunction of the blood-brain barrier (BBB) is closely related to neuroinflammation-mediated neurodegenerative disorders. Lipopolysaccharide (LPS), an endotoxin, can cause inflammation by impairing the brain endothelial barrier function and increasing the BBB permeability. Although Taraxacum coreanum NAKAI extract (TC), a traditional medicine widely used in Korea, has antioxidant and anti-inflammatory properties, the protective effects on neuroinflammation and BBB dysfunction are not fully understood. In the present study, bEnd.3 cerebral vascular endothelial cells were treated with TC followed by LPS exposure, and the effects on transendothelial electrical resistance (TEER) values, pro-inflammatory cytokine production, and expression of proteins related to inflammatory responses and tight junction integrity were assessed. The TC-treated group exhibited elevated TEER values in bEnd.3 monolayer compared to LPS-only treated group. In addition, TC treatment increased the expression of proteins involved in the tight junctions, such as ZO-1, claudin-5, and occludin. Furthermore, the TC-treated group suppressed the proteins expression-related to nuclear factor-κB (NF-κB) pathway. Taken together, TC attenuates LPS-induced neuroinflammatory responses by regulating NF-κB activation, which may contribute to protecting against BBB disruption. These findings suggest that TC may have the potential to be used as a material for functional foods to prevent neuroinflammation-related brain diseases.

Suppressing Ralstonia solanacearum and Bacterial Antibiotic Resistance Genes in Tomato Rhizosphere Soil through Companion Planting with Basil or Cilantro

The effects of companion planting on soil antibiotic resistance genes (ARGs) and associated microbial composition have remained largely unclear until now. In this study, we assessed the changes in the soil microbiome and ARGs frequencies for tomato growing soils that were companion planted with basil (TB) or cilantro (TC) using a metagenome approach. The abundance of the phytopathogen Ralstonia solanacearum was significantly lower in the TC or TB treatments compared to the tomato monoculture soils (TT). A significant enrichment of Pseudomonas and Aquabacterium and a depletion of Nocardioides and Streptomyces were observed in the TC treatment. Interestingly, both TC and TB companion planting reduced the absolute abundance and the number of subtypes of ARGs. The TC soil showed the lowest numbers of unique ARG subtypes, especially the ARGs resistant to vancomycin and rifamycin, as well as those associated with multidrug resistance. Furthermore, network analysis further revealed that Nocardioides and Streptomyces were potential hosts of ARGs, whereas Flavobacterium negatively correlated with mdtG, suggesting a suppressive effect in reducing ARGs. Together, our results suggest that the companion planting of tomatoes with basil or cilantro can reduce the risk of ARG accumulation, making it a feasible farming management tool to promote soil and plant health in sustainable agriculture.

AHNAK2 Promotes the Progression of Differentiated Thyroid Cancer through PI3K/AKT Signaling Pathway.

AHNAK2 may be used as a candidate marker for TC diagnosis and treatment. Thyroid cancer (TC) is the most frequent malignancy in endocrine carcinoma, and the incidence has been increasing for decades. To understand the molecular mechanism of DTC, we performed next-generation sequencing (NGS) on 79 paired DTC tissues and normal thyroid tissues. The RNA-sequencing (RNA-seq) data analysis results indicated that AHNAK nucleoprotein 2 (AHNAK2) was significantly upregulated in the thyroid cancer patient's tissue. We also analyzed AHNAK2 mRNA levels of DTC tissues and normal tissues from The Cancer Genome Atlas (TCGA). The association between the expression level of AHNAK2 and clinicopathological features was evaluated in the TCGA cohort. Furthermore, AHNAK2 gene expression was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) in 40 paired DTC tissues and adjacent normal thyroid tissues. The receiver operating characteristic (ROC) curve was performed to evaluate the diagnostic value of AHNAK2. For cell experiments in vitro, AHNAK2 was knocked down using small interfering RNA (siRNA), and the biological function of AHNAK2 in TC cell lines was investigated. The expression of AHNAK2 was significantly upregulated in both the TCGA cohort and the local cohort. The analysis results of the TCGA cohort indicated that the upregulation of AHNAK2 was associated with tumor size (P < 0.001), lymph node metastasis (P < 0.001), and disease stage (P < 0.001). The area under the curve (AUC, TCGA: P < 0.0001; local validated cohort: P < 0.0001) in the ROC curve revealed that AHNAK2 might be considered a diagnostic biomarker for TC. The knockdown of AHNAK2 reduced TC cell proliferation, colony formation, migration, invasion, cell cycle, and induced cell apoptosis. Furthermore, the protein levels of phospho-PI3 Kinase p85 and phospho-AKT were downregulated in the transfected TC cell. Our study results indicate that AHNAK2 may promote metastasis and proliferation of thyroid cancer through PI3K/AKT signaling pathway. Thus, AHNAK2 may be used as a candidate marker for TC diagnosis and treatment.

Protective effects of Tinospora cordifolia miers extract against hepatic and neurobehavioral deficits in thioacetamide-induced hepatic encephalopathy in rats via modulating hyperammonemia and glial cell activation

Ethnopharmacological relevanceTinospora cordifolia (TC) a potential medicinal herb, has been ethnobotanically used as an eco-friendly supplement to manage various diseases, including cerebral fever. Earlier studies have shown that TC exhibits diverse beneficial effects, including hepatoprotective and neuroprotective effects. However, the effects of TC remain unexplored in animal models of encephalopathy including hepatic encephalopathy (HE). Aim of the studyTo evaluate the effects of TC stem extract against thioacetamide (TAA)-induced behavioural and molecular alterations in HE rats. Methods and materialsThe extract was preliminarily screened through phytochemical and HR-LC/MS analysis. Animals were pre-treated with TC extract at doses 30 and 100 mg/kg, orally. Following 7 days of TC pre-treatment, HE was induced by administering TAA (300 mg/kg, i. p. thrice). Behavioural assessments were performed after 56 h of TAA first dose. The animals were then sacrificed to assess biochemical parameters in serum, liver and brain. Liver tissue was used for immunoblotting and histological studies to evaluate inflammatory and fibrotic signalling. Moreover, brain tissue was used to evaluate brain edema, activation of glial cells (GFAP, IBA-1) and NF-κB/NLRP3 downstream signalling via immunoblotting and immunohistochemical analysis in cortex and hippocampus. ResultsThe pre-treatment with TC extract effective mitigated TAA-induced behavioural alterations, lowered serum LFT (AST, ALT, ALP, bilirubin) and oxidative stress markers in liver and brain. TC treatment significantly modulated hyperammonemia, cerebral edema and preserved the integrity of BBB proteins in HE animals. TC treatment attenuated TAA-induced histological changes, tissue inflammation (pNF-κB (p65), TNF-α, NLRP3) and fibrosis (collagen, α-SMA) in liver. In addition, immunoblotting analysis revealed TC pre-treatment inhibited fibrotic proteins such as vimentin, TGF-β1 and pSmad2/3 in the liver. Our study further showed that TC treatment downregulated the expression of MAPK/NF-κB inflammatory signalling, as well as GFAP and IBA-1 (glial cell markers) in cortex and hippocampus of TAA-intoxicated rats. Additionally, TC-treated animals exhibited reduced expression of caspase3/9 and BAX induced by TAA. ConclusionThis study revealed promising insights on the protective effects of TC against HE. The findings clearly demonstrated that the significant inhibition of MAPK/NF-κB signalling and glial cell activation could be responsible for the observed beneficial effects of TC in TAA-induced HE rats.

Effect of feeding toy and the presence of a dog owner during the feeding time on dog welfare

Background and Aim: A conventional feeding bowl is the primary method that dog owners use to feed their dogs, but this may not encourage natural behaviors and may even exacerbate unwanted behaviors. This study aimed to compare a conventional feeding bowl to a feeding toy in relation to behavior, cortisol levels, and heart rate variability (HRV). Materials and Methods: The behaviors of four dogs were recorded and analyzed while being fed using either a stainless bowl (B) or a feeding toy (T) and either alone (A) or accompanied by a dog owner (O) for 30 min with each treatment (BA, BO, TA, and TO treatments). The dogs that were fed alone with the stainless bowl (BC) or the feeding toy (TC) were fed for 15 min/day for 7 days with their treatment, and serum cortisol levels measured on the first and last days of treatment. The dogs fed by the stainless bowl (BH) or the feeding toy (TH) with the owner present for 15 min for each treatment had their heart rate (HR) and HRV recorded by Polar® H10 during feedings The results were compared using a one-way analysis of variance (ANOVA), repeated measure ANOVA, and Student’s t-test. Results: The dogs spent more time eating and interacting with the feeding toys than stainless bowls. The activity of the dogs was higher when using feeding toys, particularly with the TO treatment. Cortisol levels were significantly lower on day 7 than on day 1 of the TC treatment. The dogs’ HR was higher during TH treatment than during BH treatment. All HRV parameters were decreased significantly when feeding the dog with the toys. Conclusion: The results of this study support the idea that feeding enrichment supports the natural feeding behaviors of dogs as they mimic hunting and playing behaviors. This reduced unwanted behavior, cortisol levels, and HRV, and increased food consumption, eating duration, and active behaviors. The presence of the dog’s owner is important because it can enhance feeding and active behaviors, and feeding enrichment can improve the dog’s welfare and the dog-human relationship. Keywords: cortisol, dog behavior, dog welfare, dog-human relationship, feeding toy, heart rate variability.

Effect of feeding toy and the presence of a dog owner during the feeding time on dog welfare.

A conventional feeding bowl is the primary method that dog owners use to feed their dogs, but this may not encourage natural behaviors and may even exacerbate unwanted behaviors. This study aimed to compare a conventional feeding bowl to a feeding toy in relation to behavior, cortisol levels, and heart rate variability (HRV). The behaviors of four dogs were recorded and analyzed while being fed using either a stainless bowl (B) or a feeding toy (T) and either alone (A) or accompanied by a dog owner (O) for 30 min with each treatment (BA, BO, TA, and TO treatments). The dogs that were fed alone with the stainless bowl (BC) or the feeding toy (TC) were fed for 15 min/day for 7 days with their treatment, and serum cortisol levels measured on the first and last days of treatment. The dogs fed by the stainless bowl (BH) or the feeding toy (TH) with the owner present for 15 min for each treatment had their heart rate (HR) and HRV recorded by Polar® H10 during feedings The results were compared using a one-way analysis of variance (ANOVA), repeated measure ANOVA, and Student's t-test. The dogs spent more time eating and interacting with the feeding toys than stainless bowls. The activity of the dogs was higher when using feeding toys, particularly with the TO treatment. Cortisol levels were significantly lower on day 7 than on day 1 of the TC treatment. The dogs' HR was higher during TH treatment than during BH treatment. All HRV parameters were decreased significantly when feeding the dog with the toys. The results of this study support the idea that feeding enrichment supports the natural feeding behaviors of dogs as they mimic hunting and playing behaviors. This reduced unwanted behavior, cortisol levels, and HRV, and increased food consumption, eating duration, and active behaviors. The presence of the dog's owner is important because it can enhance feeding and active behaviors, and feeding enrichment can improve the dog's welfare and the dog-human relationship.

Association of physical activity with overall mortality among long-term testicular cancer survivors: A longitudinal study.

Physical activity (PA) has been associated with reduced mortality among cancer survivors, but no study has focused on testicular cancer survivors (TCSs). We aimed to investigate the association of PA measured twice during survivorship with overall mortality in TCSs. TCSs treated during 1980 to 1994 participated in a nationwide longitudinal survey between 1998 to 2002 (S1: n = 1392) and 2007 to 2009 (S2: n = 1011). PA was self-reported by asking for the average hours per week of leisure-time PA in the past year. Responses were converted into metabolic equivalent task hours/week (MET-h/wk) and participants were categorized into: Inactives (0 MET-h/wk), Low-Actives (2-6 MET-h/wk), Actives (10-18 MET-h/wk) and High-Actives (20-48 MET-h/wk). Mortality from S1 and S2, respectively, was analyzed using the Kaplan-Meier estimator and Cox proportional hazards models until the End of Study (December 31, 2020). Mean age at S1 was 45 years (SD 10.2). Nineteen percent (n = 268) of TCSs died between S1 and EoS, with 138 dying after S2. Compared to Inactives at S1, the mortality risk among Actives was 51% lower (HR 0.49, 95% CI: 0.29-0.84) with no further mortality reduction among High-Actives. At S2, the mortality risk was at least 60% lower among the Actives, High-Actives and even the Low-Actives compared to the Inactives. Persistent Actives (≥10 MET-h/wk at S1 and S2) had a 51% lower mortality risk compared to Persistent Inactives (<10 MET-h/wk at S1 and S2; HR 0.49, 95% CI: 0.30-0.82). During long-term survivorship after TC treatment, regular and maintained PA were associated with an overall mortality risk reduction of at least 50%.

Effects of Combined Use of Olive Mill Waste Compost and Sprinkler Irrigation on GHG Emissions and Net Ecosystem Carbon Budget under Different Tillage Systems.

Traditional rice (Oryza sativa L.) production by flooding is a source of greenhouse gases (GHG), especially methane. The high consumption of water, as well as the chemical and physical degradation caused by these traditional practices in rice soils, is promoting a decrease in rice production in the Mediterranean area. The aim of this study was to monitor GHG emissions and the net ecosystem carbon balance (NECB) from rice produced with sprinkler irrigation techniques and also assess the impact of olive mill waste compost (C-OW) application and tillage on GHG emissions and the NECB. A field experiment for irrigated rice production was implemented by considering four different treatments: (1) tillage (T); (2) no tillage-direct seeding techniques (DS); (3) application of C-OW followed by tillage (TC); and (4) application of C-OW followed by direct seeding (DSC). The C-OW was only applied in the first year at a dose of 80 Mg ha-1. GHG emissions were monitored over three years in these four treatments in order to estimate the direct (first year) and residual (third year) effects of such practices. The application of C-OW caused an increase of 1.85 times the emission of CO2-C in the TC-DSC compared to the T-DS in the first year. It is noteworthy that the TC treatment was the only one that maintained an emission of CO2-C that was 42% higher than T in the third year. Regardless of the treatments and year of the study, negative values for the cumulative CH4 were found, suggesting that under sprinkler irrigation, CH4 oxidation was the dominant process. A decrease in N2O emissions was observed under direct seeding relative to the tillage treatments, although without significant differences. Tillage resulted in an increase in the global warming potential (GWP) of up to 31% with respect to direct seeding management in the third year, as a consequence of the greater carbon oxidation caused by intensive tillage. DS presented a positive NECB in the accumulation of C in the soil; therefore, it provided a greater ecological benefit to the environment. Thus, under Mediterranean conditions, rice production through a sprinkler irrigation system in combination with direct seeding techniques may be a sustainable alternative for rice crops, reducing their GWP and resulting in a lower carbon footprint. However, the use of C-OW as an organic amendment could increase the GHG emissions from rice fields irrigated by sprinklers, especially under tillage conditions.

Clinicopathological and surgical comparisons of differentiated thyroid cancer between China and the USA: A multicentered hospital-based study.

BackgroundThyroid cancer (TC), was the fastest-rising tumor of all malignancies in the world and China, predominantly differentiated thyroid cancer (DTC). However, evidence on TC stage distribution and influencing factors of late-stage were limited in China.MethodsWe carried out a retrospective study and enrolled TC patients who were first diagnosed and hospitalized in 8 hospitals in China in 2017. Logistic regression was used to evaluate associations between influencing factors and DTC stage. We extracted eligible primary DTC records newly diagnosed in 2017 from the USA's Surveillance, Epidemiology, and End Results (SEER) database. We compared clinicopathological features and surgical treatment between our DTC records and those from the SEER database.ResultsA total of 1970 eligible patients were included, with 1861 DTC patients with known stage. Among patients ≥45 years old, males (OR = 1.76, 95%CI 1.17–2.65) and those with new rural cooperative medical scheme insurance (NCMS) (OR = 1.99, 95%CI 1.38–2.88) had higher risks of late-stage DTC (stage III-IV). Compared with SEER database, over-diagnosis is more common in China [more DTC patients with onset age< 45 years old (50.3 vs. 40.7%, P < 0.001), with early-stage (81.2 vs. 76.0%, P < 0.001), and with tumors<2cm (74.9 vs. 63.7%, P < 0.001)]. Compared with the USA, TC treatment is more conservative in China. The proportion of lobectomy in our database was significantly higher than that in the SEER database (41.3 vs. 17.0%, P < 0.001).ConclusionsUnique risk factors are found to be associated with late-stage DTC in China. The differences in the aspect of clinicopathological features and surgical approaches between China and the USA indicate that potential over-diagnosis and over-surgery exist, and disparities on surgery extent may need further consideration. The findings provided references for other countries with similar patterns.

Effect of Sheep Grazing, Stocking Rates and Dolomitic Limestone Application on the Floristic Composition of a Permanent Dryland Pasture, in the Montado Agroforestry System of Southern Portugal.

Simple SummaryThe Montado is a characteristic ecosystem of the Mediterranean region, where agricultural activities, animal production and forestry coexist alongside tourism, hunting and leisure activities. Animal grazing is fundamental for the conservation of the Montado, and it is imperative to clearly understand its interactions with the pasture floristic composition (PFC) of the Montado. The objective of this study was to evaluate the effect of sheep grazing, stocking rates and dolomitic limestone application on the floristic composition of permanent dryland pastures, in the Montado agroforestry system of Southern Portugal. The type of grazing influences the PFC, which may be positively or negatively impacted, depending on the adopted system. Deferred grazing seems to benefit the disappearance of undesirable plants and the appearance of desired plants. The results of this study allow for more informed management decisions and a potential increase in animal production but also improve the knowledge of conservation strategies in the Montado.The Montado is a complex agroforestry–pastoral ecosystem due to the interactions between soil–pasture–trees–animals and climate. The typical Montado soil has an acidic pH and manganese toxicity, which affect the pasture’s productivity and pasture floristic composition (PFC). The PFC, on the other hand, can also be influenced by the type and intensity of grazing, which can lead to significant decreases in the amount of biomass produced and the biodiversity of species in the pasture. The objective of this study was to evaluate the effect of grazing type, by sheep, and different stocking rates on the PFC throughout the vegetative pasture cycle in areas with and without dolomitic limestone application. Thus, four treatments (P1UC to P4TC) were constituted: P1UC—without limestone application (U) and continuous grazing (CG); P2UD—U and deferred grazing (DG); P3TD—with the application of limestone (T) and DG; P4TC—T and CG. In DG plots, the placement and removal of the animals were carried out as a function of the average height of the pasture (placement—10 cm; removal—3 to 5 cm). The PFC was characterized in winter, at the peak of spring and in late spring. The PFC data were subjected to a multilevel pattern analysis (ISA). The combination of rainfall and temperature influenced the pasture growth rates and consequently the height of the pasture at different times of the year. Therefore, with the different growth rates of the pasture throughout the year, the sheep remain for different periods of time in the deferred grazing treatments. In the four treatments, 103 plant species were identified. The most representative botanical families in the four treatments were Asteraceae, Fabaceae and Poaceae. ISA identified 14 bioindicator species: eight for the winter period, three for the late spring vegetative period and three for the TC treatment.

Physical modification of maize starch by gelatinizations and cold storage

The effects of gelatinization at three selected temperatures (DSC characteristic peaks temperature: TO, TP, and TC) and subsequent cold storage (CS) treatment on structural characteristics, pasting, and rheological properties of maize starch (MS) were investigated. The pasting, rheological properties of MS was changed with the increase of gelatinization temperature from TO to TC, but were not further significantly changed if the gelatinization temperature was higher than TC. Pasting and thermal properties analysis suggested that gelatinization at TC (TC treatment) significantly increased the gelatinization and pasting temperature of MS. Moreover, TC treatment decreased breakdown viscosity by 8.49 times and setback viscosity by 2.53 times. Dynamic rheological measurements revealed that the TC treatment caused the lower G' and G" of MS, and decreased the thickening coefficient by 55.17 %. These results indicated that TC treatment could enhance the thermal stability properties of MS, inhibiting the shear and short-term retrogradation, the shear-thinning behavior of MS. Interestingly, the CS treatment further inhibited the shear and short-term retrogradation and the shear-thinning behavior of MS. The leaked starch molecules aggregate to form a harder structure after gelatinization and starch molecules were further aggregated after CS treatment, these all were hypothesized to be responsible for these results.

Effects of plastic mulching on soil CO2 efflux in a cotton field in northwestern China

In Northwestern China, more and more traditional cultivation system (TC) with no mulching and flood irrigation have been replaced by modern cultivation technology (MC) combining plastic film mulching with drip irrigation. Does plastic film mulching increase or reduce soil CO2 emission in arid areas? In order to study the effects of plastic mulching on soil CO2 efflux, a field study was conducted to compare soil CO2 concentration, soil CO2 efflux, soil temperature and moisture between the TC treatment and the MC treatment during a cotton growing season in Northwestern China. The seasonal patterns of soil profile temperature and soil moisture in the TC treatment were similar to that in the MC treatment. The mean value of soil profile temperature in the MC treatment was higher than that in the TC treatment. Except for soil moisture at 15 cm depth, the mean value of soil moisture at 5 cm and 10 cm depths in the MC treatment was higher than that in the TC treatment. The variation patterns of soil CO2 concentration and soil CO2 efflux in MC treatment were different to that in the TC treatment. Although the peak of soil CO2 concentration in the TC treatment was earlier than that in the MC treatment, the duration of soil CO2 concentration with high values in TC treatment was shorter than that in the MC treatment. Based on the model of Fick’s first diffusion law, soil surface CO2 efflux in the MC and TC treatments were determined. The surface CO2 efflux in the TC treatment calculated by Fick’s first diffusion law model was in good agreement with the value measured by chamber method. The seasonal curve of soil surface CO2 efflux in the MC treatment indicate the similar pattern with that in the TC treatment, and the rate of CO2 efflux was lower in the MC system. In the MC treatment, the seasonal variation of soil surface efflux was explained more by soil moisture than by soil temperature. However, in the TC treatment, the seasonal variation of soil surface efflux was explained more by soil temperature than by soil moisture. Over the completely experimental period, accumulated rates of soil CO2 efflux were 361 g C m−2 and 474 g C m−2 for the MC and TC system, respectively. We concluded that converting agricultural practices from traditional cultivation to the plastic mulching cultivation could reduce soil CO2 efflux by approximately 110 g C m−2 year−1 in agricultural land in arid areas of Northwestern China.

Effect of Post-grazing Sward Height on Ingestive Behavior, Dry Matter Intake, and Milk Production of Holstein Dairy Cows

Sward height is strongly related to the daily dry matter intake of grazing dairy cows, which consequently determines animal performance. Despite that, few studies have explored the potential to increase milk production by managing post-grazing sward height. An experiment was carried out to evaluate the effect of three defoliation intensities on a Lolium arundinaceum-based pasture on frequency and length of grazing meals and ruminating bouts, daily grazing and ruminating time, feeding stations and patches exploration, and dry matter intake and milk production of dairy cows. The treatments imposed were three different post-grazing sward heights: control (TC), medium (TM), and lax (TL), which were managed with 6, 9, and 12 cm of post-grazing sward heights during autumn and winter, and 9, 12, and 15 cm of post-grazing sward heights during spring, respectively. Thirty-six autumn-calving Holstein cows were grouped by parity (2.6 ± 0.8), body weight (618 ± 48) kg, and body condition score (2.8 ± 0.2) and randomized to the treatments. The pasture was accessed from 08:00 to 14:00 and 17:00 to 03:00 during spring and no supplement was involved during the evaluation period. Daily grazing time averaged 508 ± 15 min and was not affected by treatment. The reduction of post-grazing sward height increased the length of the first grazing session in the morning and the afternoon. The number of grazing sessions was greater on TL than on TM, with no difference in TC. The number of feeding stations (the hypothetical semi-circle in front of an animal from which the bites were taken without moving the front forefeet) visited was less on TC than on TL, and neither of them differed from TM. Dry matter (DM) intake was lesser on TC than on TM and TL (14.7 vs. 17.8 kg DM). Milk production during the evaluation period was 13.1, 16.2, and 18.7 kg/day for TC, TM, and TL, respectively. The milk fat, protein, and lactose content did not differ between treatments. The cows on TC exhibited a lower intake rate, although they were less selective, probably as a consequence of the sward structure of TC treatment. The cows on TM adopted a compensation mechanism which allowed them to achieve the same dry matter intake as cows on TL, but lower milk production. The cows on TL were more selective than TC and TM resulting in higher digestible dry matter intake and consequently higher milk production. The intensity of defoliation impacts on the animal-plant interaction, and constitutes a valuable management tool that can be used to boost forage intake and milk production. The new developments on information technology would allow researchers to link behavioral data with response variables (e.g., milk production, health, welfare, etc.) at different spatio-temporal scales and support short and long-term management decisions.

ALDH5A1 acts as a tumour promoter and has a prognostic impact in papillary thyroid carcinoma.

Thyroid cancer is the most common endocrine carcinoma, with papillary thyroid carcinoma (PTC) accounting for 80%-90% of thyroid cancers. Accumulating studies reported that mitochondria plays an important role in the regulation of cell proliferation. ALDH5A1, may function as an oncogene or tumour suppressor in various human cancers, and the role of ALDH5A1 in PTC is still unclear. The aim of this study was to investigate the clinical significance of ALDH5A1 expression and its functions in PTC. In this present study, we studied ALDH5A1 expression on primary papillary thyroid carcinoma (PTC) in The Cancer Genome Atlas (TCGA) database. Results showed that the levels of ALDH5A1 were found positively correlated with tumour stage, metastasis, lymph node stage, and higher levels of ALDH5A1 demonstrated poor disease-free survival (DFS). Immunohistochemistry (IHC) revealed that significantly higher expression of ALDH5A1 was found in PTC tissues. On the other hand, knockdown of ALDH5A1 significantly inhibited PTC cell proliferation, migration and invasion detection found the migration and invasion of cells also were hindered when ALDH5A1 level was reduced. The knockdown of ALDH5A1 inhibited the expression of Vimentin and promoted the expression of E-cadherin. In brief, knockdown of ALDH5A1may act as a novel molecular target for the prevention and treatment of PTC. SIGNIFICANCE OF THE STUDY: The present study focused on the role and the potential mechanism of ALDH5A1 in papillary thyroid carcinoma. We demonstrated that reduced expression of ALDH5A1 might inhibit the progression of TC by inhibiting cell proliferation, migration and invasion and reversing epithelial-mesenchymal transition (EMT). The findings ensured the interaction relation between ALDH5A1 and EMT in PTC, providing a novel biological marker for PTC and enriching the potential strategies for TC treatment.

Abstract 2044: Chemotherapy induced neutropenia in rats following administration of eflapegrastim or pegfilgrastim on the same day at three different timepoints and at 24 hours post-chemotherapy

Abstract Background: Eflapegrastim is a long-acting G-CSF that is comprised of a G-CSF analog and a recombinant IgG4 Fc fragment conjugated at their N-termini via a short polyethylene glycol linker. Currently, G-CSF products are administered 24 hours after chemotherapy. We compared the duration of neutropenia (DN) after treatment with eflapegrastim and pegfilgrastim given on the same-day, at three different timepoints, and after 24 hours in chemotherapy-induced neutropenic rats. Method: Neutropenia was induced in rats via intraperitoneal administration of cyclophosphamide and docetaxel (TC) chemotherapy. Rats (5 animals per group) were assigned to dose groups of either vehicle or equivalent doses (based on human clinical doses) of pegfilgrastim (103.3 µg/kg) or eflapegrastim (61.8 µg/kg as G-CSF) on the same day as chemotherapy at 0, 2, and 5 hours or 24 hours after chemotherapy. The primary endpoint was the difference in DN after TC treatment and administration of eflapegrastim or pegfilgrastim at the specified timepoints. Results: In the vehicle group, the TC regimen induced neutropenia in all animals with an ANC nadir occurring on approximately Day 4 and a mean DN of nearly 7 days. The mean DN values for treatment with eflapegrastim on the same day, at 0, 2, and 5 hours, and after 24 hours were similar and the mean DN values for eflapegrastim were significantly lower than for pegfilgrastim at all dosing timepoints (Table). Time after Chemotherapy(hours)Mean Duration of Neutropenia (days)Difference (95% CI)Eflapegrastim Time vs 24 hoursDifference (95% CI)EflapegrastimPegfilgrastim00.62.2-1.6 (-3.067, -0.133)0.4 (-0.08, 1.28)20.21.8-1.6 (-2.84, -0.36)0 (-0.19, 0.59)50.01.2-1.2 (-1.20, -1.20)-0.2 (0, 0)240.21.8-1.6 (-2.84, -0.36)NA Conclusions: The DN for same-day treatment with eflapegrastim at 0, 2, and 5 hours was similar to that of the treatment after 24-hours. However, the mean DN values were significantly lower for eflapegrastim than pegfilgrastim at all time points. Further exploration of the potential advantage of same-day dosing with eflapegrastim in patients undergoing chemotherapy is warranted. Citation Format: Yu-Yon Kim, Eunjung Kim, Hyesun Han, Seungjae Baek, Shanta Chawla, Prasad Kolli, Gajanan Bhat, Francois Lebel. Chemotherapy induced neutropenia in rats following administration of eflapegrastim or pegfilgrastim on the same day at three different timepoints and at 24 hours post-chemotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2044.

DESENVOLVIMENTO DE CITRONELA (CYMBOPOGON WINTERIANUS) IRRIGADA COM ESGOTO DOMÉSTICO TRATADO E COM ÁGUA DE ABASTECIMENTO

Este trabalho teve como objetivo avaliar o desenvolvimento de capim citronela (Cymbopogon winterianus) irrigado com água de abastecimento fornecida pela concessionária (TA), esgoto doméstico tratado, após a adição de cloro (TB), e esgoto doméstico tratado, antes da adição de cloro (TC). Utilizou-se o efluente de uma estação de tratamento de esgoto (ETE) do tipo reator anaeróbio de fluxo ascendente (RAFA). A irrigação, com cada efluente, foi realizada em sete vasos, cada vaso com duas mudas do vegetal, totalizando14 plantas. Analisaram-se: índice de clorofila nas folhas (IC); teor de massa seca (MS); teor de massa inerte (MI). Para o IC, os resultados, em Soil Plant Analysis Development (SPAD), foram: 18 (TA), 31,73 (TB) e 37,03 (TC). Para o MS, os resultados, em %m/m, foram: 86,14 (TA), 83,75 (TB) e 82,18 (TC). Para o MI, os resultados, em %m/m, foram: 2,25 (TA), 1,71 (TB) e 1,63 (TC). Percebeu-se que plantas regadas com efluentes tratados apresentaram visualmente um melhor desenvolvimento foliar e tonalidade mais escura deverde. O efluente oriundo do tratamento TC proporcionou a maior produção de clorofila, indicando um maior potencial de aproveitamento. O tratamento TA proporcionou a produção de maiores quantidades de massa seca e de massa inerte.

Azacytidine Sensitizes AML Cells for Effective Elimination By CD123 CAR T-Cells

Chimeric antigen receptor T-cells (CAR Tc) have yielded impressive remission rates in treatment-refractory B-cell malignancies (B-ALL and B-lymphomas) by targeting CD19, resulting in the first FDA approved CAR Tc therapies, Kymriah and Yescarta. However, the translation of these results for other cancer entities remains a challenge. Pre-clinical studies using second-generation CAR Tc against the interleukin-3 receptor alpha chain (CD123) engendered strong anti-leukemic activity. CD123 CAR Tc clinical studies resulted in transient responses, or complete remission but at the expense of on-target off-tumor toxicities. Our studies employing third-generation anti-CD123 CAR Tc demonstrate strong anti-leukemic activity with no adverse effects in vivo. However, the leukemia was not completely eradicated. Combining anti-CD123 CAR Tc with DNA hypomethylating (HMA) agents may enhance the anti-leukemic effect and survival. HMAs such as azacytidine (Aza) activate key epigenetically silenced pathways in AML cells, inhibiting cell proliferation while enhancing cell immunogenicity. We hypothesized that Aza will increase the expression of CD123 on AML cells resulting in long-term disease eradication by anti-CD123 CAR Tc. The anti-leukemic efficacy, survival advantage, safety and feasibility of the combination treatment with Aza and anti-CD123 CAR Tc were evaluated in vivo. HL-60 (CD123med), MLL-2 (CD123lo), MOLM-13 (CD123hi), primary de novo and relapsed/refractory (r/r) AML cells were cultured for 0-8 days in the presence of Aza (0µM-5µM) and analysed for their CD123 expression by flow cytometry, quantitative western blot and RNAseq. The anti-CD123 CAR was constructed with the humanized CSL362-based ScFv and the CD28-OX40-CD3ζ signaling domain, encoded in a third-generation lentiviral vector and expressed in CD3+ Tc from healthy donors. Rag2γc-/- mice (n=12-16/ group) were engrafted with 1x105 MOLM13/ffLuc AML cells and treated with PBS, 5x106 Non-transduced (NTD) Tc orCAR Tc, 4x 2.5mg/kg Aza, or 5x106 CAR Tc following 4x Aza (2.5mg/kg). Leukemic burden was assessed weekly by bioluminescence imaging. Tc activity and immunophenotyping was performed using flow cytometry at day 35 post engraftment, and survival was monitored. HL-60, MLL-2 and MOLM-13 cells showed significant increases in HLA-DR, PD-L1, STAT1 and IRF7 expression, as well as CD123 when exposed to Aza (Fig 1A,B). Interestingly, the increased effect was seen from day one regardless of concentration. This was similarly reflected in AML patient cells. Aza treatment also arrested cell proliferation and decreased viability in both cell lines and patient cells suggesting Aza can aid in the anti-leukemic effect. Rag2γc-/- mice engrafted with MOLM-13 and treated with Aza and CD123 CAR Tc demonstrated suppressed growth, and eradication of MOLM-13 cells compared to mice treated with CD123 CAR Tc or Aza alone. Additionally, a significant decrease in residual CD123+ cells in the bone marrow (BM) of dual treated mice was seen (Fig 1C). A higher frequency of residual CD8+ T-cells in the BM, and CD4+ Tc in the peripheral blood (PB) and BM of dual treated mice was observed compared to CAR Tc only treated mice. Most prominently, we found a significantly higher mean number of stem cell-like and central memory CD8+ Tc in the BM of dual treated mice (232 cells/µl and 208cells/µl, respectively) compared to the CAR Tc only group (55 cells/µl and 23 cells/µl, respectively). Assessment of immune checkpoint markers on residual CAR Tc of dual treated mice revealed significantly decreased levels of CTLA-4, PD-1 and TIM-3 in the BM, and CTLA-4 in the PB compared to the CAR Tc only group. While CAR Tc treatment alone demonstrated a survival advantage compared to PBS, NTD or Aza treated mice, Aza and CAR Tc treatment had a significantly higher survival rate compared to the CAR Tc only group (92% vs. 46% at day 50, p&lt;.01). Our findings indicate that Aza increases immunogenicity and augments the cell surface expression of CD123 on AML cells, allowing enhanced recognition and elimination of malignant cells by CD123 CAR Tc. This is the first demonstration that HMAs and CAR Tc immunotherapy can be used synergistically to treat AML. Considering HMAs are currently under clinical investigation in AML, our data encourage further clinical evaluation of this dual treatment in r/r AML, including high-risk patients that are chemotherapy or allogeneic transplantation ineligible. Disclosures Hughes: Novartis, Bristol-Myers Squibb, Celgene: Research Funding; Novartis, Bristol-Myers Squibb: Consultancy, Other: Travel. White:BMS: Honoraria, Research Funding; AMGEN: Honoraria, Speakers Bureau. Yong:Novartis: Honoraria, Research Funding; Celgene: Research Funding; BMS: Honoraria, Research Funding.

Highly bioavailable curcumin powder suppresses articular cartilage damage in rats with mono-iodoacetate (MIA)-induced osteoarthritis.

This study was performed to investigate the effects of highly bioavailable curcumin as Theracurmin® (TC) in rats with monosodium iodoacetate (MIA)-induced osteoarthritis (OA). Seventy-seven male Wistar rats were divided into six groups: normal, negative control (MIA only), positive control (Cerebrex), and three experimental groups treated with 500, 1300, or 2600mg/kg of TC for 5weeks. MIA injection-induced OA caused 30% weight-bearing imbalance whereas weight bearing imbalance was significantly improved in the TC groups. Mankin scores revealed TC treatment had significantly ameliorated cartilage damage and chondrocyte decrease. The expressions of nitrotyrosine, tumor necrosis factor-α, phosphorylated nuclear factor kappa B cells, and cleaved caspase-3 were markedly increased in rat with MIA-induced OA, but the TC-treated groups exhibited a significant reduction in the number of immunoreactive cells in a dose-dependent manner. In conclusion, administration of TC contributes to the anti-arthritic effect in rat with MIA-induced OA.

Abstract 282: Minnelide improves the efficacy of the nab-paclitaxel plus gemcitabine plus cisplatin chemotherapy in mouse models of PDAC

Abstract Background: There have been some recently developed regimens which have improved the survival of patients with advanced pancreatic cancer. In a recent Phase Ib/II pilot trial in patients with advanced stage IV PDAC, nab-paclitaxel/gemcitabine/cisplatin regimen (triple combination, TC) exhibited promising clinical activity with a response rate of 71% and overall median survival of 16.5 months associated with manageable adverse effects (Jameson, et al., J. Clin. Oncol. 2017 35:4 suppl, 341). Minnelide (Min), a novel water soluble pro-drug of triptolide, a potent antitumor agent isolated from a Chinese medicinal herb, has demonstrated efficacy against several human cancers implanted in mouse models. Min is currently in Phase II trial in patients with refractory pancreatic cancer. In this study we evaluated the synergy between the TC and Min in various pancreatic cancer models (including the treatment sequence of the agents). Methods: The drug combination efficacy was evaluated in athymic nude mouse models generated with MIA PaCa-2 cells and patient derived xenografts (PDX) implanted subcutaneously. The mice were treated with vehicle control, Min (QDx21), TC (QWx3) or TC (QWx3) plus Min(QDx21). In the PDX models Min (QDx21) given before or after the TC (QWx3) was also evaluated. To evaluate the effects of the combination treatment on tumor immune microenvironment, the KPC transgenic mice (K-RasG12D/+, p53R172H/+) were also treated with same combinations. Tumor volumes were assessed by ultrasound and immune cell infiltration was assessed by immunohistochemistry (IHC). Results: The TC plus Min shows the greatest effect in tumor growth inhibition which remains persistent even after the treatment is discontinued in both the cell line xenograft model and the PDX models with several of the mice exhibiting complete remissions. In the MIA PaCa-2 cell line xenograft model, comparing to the vehicle control the tumor growth inhibition after drug treatment (21 days) was 97%, 93%, and 100% for Min, TC, and Min + TC, respectively. The animal survival at 45 days was 0%, 73%, 67%, and 90% for vehicle, Min, TC, and Min + TC, respectively. In the PDX model, the tumor growth inhibition after drug treatment was 86%, 84%, and 96% comparing to the vehicle, whereas the animal survival at 45 days was 0%, 89%, 100% and 100% for vehicle, Min, TC, and Min + TC, respectively. Min administered prior to or after the TC treatment also exhibited improved efficacy versus TC alone, with tumor growth inhibition similar to those treated with the agents concurrently. These results provide preclinical insights for the design of clinical trials that may further improve outcome in patients with advanced PDAC. (This work was supported by the SU2C-CRUK-Lustgarten Dream Team grant (Grant number: SU2C-AACR-DT-20-16)) Citation Format: Liang Ng, Pawan Noel, Stephanie Mou, Daniel D. Von Hoff, Haiyong Han. Minnelide improves the efficacy of the nab-paclitaxel plus gemcitabine plus cisplatin chemotherapy in mouse models of PDAC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 282.

Chemotherapy and trastuzumab-induced cardiotoxicity in geriatric breast cancer patients.

e23043 Background: Cardiotoxicity is a known side effect of anthracyclines &amp; trastuzumab. Dilated cardiomyopathy is the most common form of cardiotoxicity associated with this treatment. Cardiovascular disease &amp; cancer are common conditions in the geriatric populations. These comorbidities complicate the treatment of geriatric breast cancer patients. The purpose of this study was to determine the cardiotoxic effects of chemotherapy &amp; trastuzumab in a geriatric population of breast cancer patients. Methods: An institutional database of a total of 269 patients with histologically confirmed invasive or in-situ breast cancer with age 65 years or older at the time of diagnosis was reviewed in an IRB approved fashion. Patients were divided in groups depending on the therapy (AC/TAC, CMF, TC and Trastuzumab) they received. Fisher’s exact test was used to calculate overall survival (OS). Results: Of the 269 patients, 72 (26.8%) were offered chemotherapy &amp; 197 (73.2%) were not. Of the patients offered chemotherapy, 58 (21.6%) accepted treatment &amp; 14 (5.2%) declined. 17 (6.7%) had an early termination, AC n = 11,( 4.1%), TAC n = 13, (4.8%), CMF n = 0, (0%), Trastuzumab n = 12, (4.5%), TC n = 11, (4.1%). Cardiovascular risk factors taken into consideration were, CAD n = 30, (13.8%), HLD n = 23, (10.6%), HTN n = 98, (45.2%), HTN/HLD n = 66, (30.4%), DM (N = 45, (16.7%), Smoking n = 121, (46.4%) &amp; BMI &gt; 30, n = 116,( 48.1%). Patients completing chemotherapy had higher OS than patients who had early termination (P = 0.017). Of the 24 patients who received Anthracycline containing regimen, 1 experienced cardiotoxicity. There was no statistically significant difference in OS between AC, TAC &amp; Trastuzumab (P = 0.570, P = 0.547, P = 0.614). A decrease in OS was seen for patient who received TC (P = 0.002). Of the 12 who received trastuzumab, 1 experienced cardiotoxicity. There was a significant association between patients with a high BMI ( &gt; 30) &amp; increased cardiovascular comorbidities (P = 0.0009) but had no impact early on termination of chemotherapy or OS, NS (P = 0.7). Conclusions: The cardiotoxic effect of anthracyclines and trastuzumab are well known, however in our study, only 2 elderly breast cancer patients who received chemotherapy experienced cardiotoxicity. As would be expected, those who completed chemotherapy had improved OS. There were no difference in OS between AC, TAC and Trastuzumab, however TC treatment was associated with a decrease in OS.BMI had an impact on cardiovascular comorbidities but not on early termination of chemotherapy or OS.