Lipid Reduction Research Articles

Leaf gas exchange and bean quality fluctuations over the whole canopy vertical profile of Arabic coffee cultivated under elevated CO2

Leaves in different positions respond differently to dynamic fluctuations in light availability, temperature and to multiple environmental stresses. The current hypothesis states that elevated atmospheric CO2 (e[CO2]) can compensate for the negative effects of water scarcity regarding leaf gas exchanges and coffee bean quality traits over the canopy vertical profile, in interactions with light and temperature microclimate during the two final stages of berry development. Responses of Coffea arabica L. were observed in the 5th year of a free air CO2 enrichment experiment (FACE) under water-limited rainfed conditions. The light dependent leaf photosynthesis curves (A/PAR) were modelled for leaves sampled from vertical profile divided into four 50-cm thick layers. e[CO2] significantly increased gross photosynthesis (AmaxGross), the apparent quantum yield efficiency, light compensation point, light saturation point (LSP) and dark respiration rate (Rd). As a specific stage response, considering berry ripening, all parameters calculated from A/PAR were insensitive to leaf position over the vertical profile. Lack of a progressive increase in AmaxGross and LSP was observed over the whole canopy profile in both stages, especially in the two lowest layers, indicating leaf plasticity to light. Negative correlation of Rd to leaf temperature (TL) was observed under e[CO2] in both stages. Under e[CO2], stomatal conductance was also negatively correlated with TL, reducing leaf transpiration and Rd even with increasing TL. This indicated coffee leaf acclimation to elevated temperatures under e[CO2] and water restriction. The e[CO2] attenuation occurred under water restriction, especially in A and water use efficiency, in both stages, with the exception of the lowest two layers. Under e[CO2], coffee produced berries in moderate- and high light level layers, with homogeneous distribution among them, contrasted to the heterogeneous distribution under actual CO2. e[CO2] led to increased caffeine content in the highest layer, with reduction of chlorogenic acid and lipids under moderate light and to raised levels of sugar in the shaded low layer. The ability of coffee to respond to e[CO2] under limited soil water was expressed through the integrated individual leaf capacities to use the available light and water, resulting in final plant investments in new reproductive structures in moderate and high light level layers.

Aerobic Training Impacts on Blood Cholesterol of Women with Gestational Diabetes

Gestational diabetes mellitus has been identified as a major complication of pregnancies and has remained a major cause of perinatal morbidity and mortality, as well as maternal morbidity. Lipid abnormalities significantly contribute to the increased risk of cardiovascular disease and other morbidities in diabetics. One of the benefits of regular exercise is a reduction in cardiovascular risk factors through the improvement of the lipid profile and reduction of hypertension. This study investigated the effect of aerobic training on the cholesterol level of women with gestational diabetes. Thirty-four (34) women who met the inclusion criteria for this study were randomized into Exercise and Control groups. Exercise group participated in structured exercise programme, 3 sessions a week for 8 weeks with each session lasting a maximum of 60 minutes while the Control group did not participate in any form of structured exercise programme for the period of the study. Data were collected at baseline, at 4th week and at 8th week into the exercise programme. Thirty (30) of the participants completed the study and so only their data were analysed using Repeated Measure ANOVA and Independent t-test. The result showed significant changes in the total cholesterol (F[2, 28]) = 268.316, P = 0.001), of the Exercise group compared to the control. Independent t-test used to compare the lipid profile of the Experimental and Control groups showed significant difference between the means of the 2 groups at week 4 (total cholesterol (t(28) 2.164, P = 0.039, triglyceride [t(28) 3.146 P = 0.004] and week 8 (total cholesterol t(28) 4.596, P = 0.001, triglyceride t(28) 5.534, P = 0.001) but not at baseline (total cholesterol t(28) -901, P = 0.375, triglyceride t(28) 1.612, P = 0.118). It was concluded that structured aerobic exercises may have positive effect on the components of lipid profile in women with gestational diabetes.

Oxidative Stress Mediated Cytotoxicity, Cell Cycle Arrest, and Apoptosis Induced by Rosa damascena in Human Cervical Cancer HeLa Cells.

Rosa damascena Mill (Damask rose), belonging to the Rosaceae family, is known for medicinal purposes in traditional medicine system. However, its anticancer activity has not been studied yet in detail. Herein, we aimed to investigate the cytotoxic effects of R. damascena hexane (RA-HE) and methanolic (RA-ME) extracts against human breast (MCF-7), lung epithelial (A-549), and cervical (HeLa) cancer cells. The RA-HE and RA-ME showed more potent cytotoxic effects against HeLa cells with an IC50 of 819.6 and 198.4 μg/ml, respectively. Further, cytotoxic concentrations of most effective extract (RA-ME) were used to evaluate the mechanism of cytotoxicity involved in HeLa cells. A concentration-dependent induction of lipid peroxidation (LPO) and reduction of glutathione (GSH) in HeLa cells treated with 250-1000 μg/ml of RA-ME confirms the association of oxidative stress. We also detected a noteworthy increase in reactive oxygen species (ROS) production and a decline in mitochondrial membrane potential (MMP) level in RA-ME-exposed HeLa cells. Flow cytometric data showed a strong dose-response relationship in cell cycle analysis between subG1 phase in HeLa cells and RA-ME treatment. Similarly, a concentration-dependent increase was recorded with Annexin V assay in HeLa cells going to late apoptosis. In conclusion, our findings suggest that RA-ME-induced cytotoxicity and apoptosis in HeLa cells are mediated by oxidative stress.

601. Disparities in Diabetes Care: Smoking Cessation among Women and Minorities Living with HIV at an Urban Academic Medical Center

BackgroundPeople living with HIV (PLWH) and diabetes mellitus are at increased risk of developing significant medical complications such as atherosclerotic cardiovascular disease. Disproportionate rates of diabetes and HIV among minority groups raise the issue of how demographic disparities may impact care. The American Diabetes Association (ADA) 2020 guidelines for diabetes care recommend optimal glycemic levels (A), blood pressure control (B), lipid reduction (C), and smoking cessation (N), commonly referred to as ABC or ABCN criteria. This quality assessment project examines diabetes management in PLWH by gender, race/ethnicity, and BMI, in a predominantly minority-serving clinic, as assessed by rates of guideline adherence to the above metrics.MethodsThis project was reviewed and approved by the Rutgers IRB. Patients from an HIV registry of University Hospital Infectious Disease Outpatient clinic in Newark, NJ were reviewed for a diagnosis of diabetes and both a clinic visit and an A1c score recorded between 2/1/2019 and 1/31/2020. Achieving glycemic target was defined as HbA1c < 7.5 for patients < 65 and HbA1c < 8 for patients > 65. Target adherence criteria also included a blood pressure average of < 140/90 over this period and an LDL-c of < 100 mg/dL. Non-smoking status includes both former and never smokers.ResultsOf 1035 patients reviewed, a total of 172 met criteria. Adherence rate for achieving goal HbA1c was 61.6% (95% CI 54.2-68.6, n=172). Blood pressure and LDL-c adherence rates were 65.1% (95% CI 57.7-71.8, n=172) and 67.4% (95% CI 60.1-74.0, n=172), respectively. ABC and ABCN rates were 24.4% (95% CI 18.6-31.4, n=172) and 18.6% (95% CI 13.5-25.1, n=172). The overall smoking rate, as well as the rates in the female subgroup, those with BMI 18.5-24.9, and the non-Hispanic black subgroup were significantly higher than the national average (P< 0.05).Table 1: Demographic Data of PLWH and Diabetes Table 2: Adherence to ABCN Criteria in Diabetes Care by Demographics for PLWH from 2/1/2019 – 1/31/2020 ConclusionFor diabetic PLWH, smoking cessation requires improvement, particularly in female, normal BMI, and non-Hispanic black subgroups. These findings, in addition to a majority overweight patient population, highlight the need for increased education and interventions aimed at nutritional counseling and risk factor mitigation among all patient subgroups.Disclosures All Authors: No reported disclosures

Insecticidal impacts of Bacillus flexus S13 biofilm'extracellular matrix' on cowpea weevil, Callosobruchus maculatus

Abstract In an attempt to throw light on the chemical composition of the extracellular matrix (ECM) of Bacillus flexes S13; biofilm’ and its potential activity as insect biocontrol agents. This study investigated the insecticidal efficacy of ECM of Bacillus flexus; S13 against adults of Callosobruchus maculatus. The toxicity results revealed LC50 values as 22.6455 g/100 ml and 10.8524 g/100 ml after 24 and 48 h respectively. Also, the Biochemical assays revealed a significant reduction in total protein, total lipids and chitin contents in survival adults treated with LC50 of Bacillus flexus' ECM where's, the total protein was decreased down to the half (15.54 mg/g) in adults treating with ECM compared to untreated-negative control (39.34 mg/g) while the positive control group treated with NaCl showed a mild decrease (23.57 mg/g). Furthermore, the GC/MS analyses of the methanolic solution of Bacillus flexus ECM showed a high content of polyphenols especially; flavonoid compounds while the major compound was (3, 5, 7, 3\, 4\, penta hydroxyl flavone)with percentage of (40.93%).

Protective effect of ethanolic extract of Actinoscirpus grossus tubers against ethanol induced liver toxicity in albino rats

Liver detoxifies majority of chemicals and plays an important role in the maintenance of homeostasis of the body. In the recent years due to overwhelming use of drugs and alcohol there is an increased incidence of liver toxicities. The tubers of Actinoscirpus grossus are used in liver disease as a liver tonic in folklore medicine; However, there is no experimental basis for its therapeutic use. Hence the present study was conducted to evaluate the hepatoprotective potentials of ethanolic extract of Actinoscirpus grossus tubers in ethanol induced hepatotoxicity in albino rats. Daily administration of 20% ethanol v/v p.o. at a dose of 5 g/kg for 30 consecutive days caused significant liver toxicity and there after maintained for 15 consecutive days with normal rat pellet and water ad libitum. The treatment groups were administered with group specific drugs for 15 consecutive days after 30 days of ethanol administration and control group rats were maintained for 15 consecutive days with normal rat pellet and water ad libitum. Liver toxicity was assessed on 45th day by estimating biochemical, histological and antioxidant level. Daily administration of 20% ethanol significantly increased liver enzymes and caused oxidative stress by increase in the lipid peroxidation and reduction in the catalase and glutathione peroxidase activity in liver homogenate. The histopathological examination revealed micro vesicular and macro vesicular fatty changes predominantly with mild periportal inflammation, cell infiltrate and areas of mild dilatation of sinusoids with ballooning degeneration of hepatocytes. Treatment with ethanolic extract of A. grossus tuber significantly restored the liver enzymes especially SGOT and attenuated elevated lipid peroxidation and restored catalase and glutathione peroxidase activity. The histopathological changes revealed significant reduction in the hepatocyte ballooning portal fibrosis. Thus, tubers of A. grossus plant may serve as reservoir of natural products having diverse functions and could be utilised as a potent hepatoprotective agent.

Macrophage-activating lipopeptide-2 and corticotropin-releasing hormone stimulate the inflammatory signalling in human sebocytes through activation of stearoyl-CoA desaturase and fatty acid desaturase 2.

The macrophage-activating lipopeptide-2 (MALP-2) activates cells carrying a functional Toll-like receptor (TLR)-2/6. Human sebocytes express functional TLR-2, TLR-4 and CD14. Upregulation of stearoyl-CoA desaturase (SCD) and fatty acid desaturase-2 (FADS2) expression induces pro-inflammatory sebaceous activity. On the other hand, corticotropin-releasing hormone (CRH) is likely to serve as an autocrine stress hormone in human sebocytes. In addition to its antiproliferative, lipogenetic and androgen-activating functions, CRH exhibits a pro-inflammatory action and its expression is upregulated in acne-involved sebaceous glands. Determination of the pro-inflammatory function of MALP-2 and CRH and clarification of the option that MALP-2 and/or CRH activity on human sebocytes might be mediated through SCD and/or FADS2. SZ95 sebocytes were treated with MALP-2, CRH and the SCD inhibitor/ligand FPCA. SCD, FADS2, TLR-2 mRNA and protein levels and IL-6 and IL-8 secretion were investigated. Intracellular CRH levels were assessed under treatment with CRH, MALP-2, linoleic acid and arachidonic acid. Phorbol 12-myristate 13-acetate and dexamethasone served as positive and negative controls, respectively. MALP-2 upregulated SCD, FADS2, TLR-2 mRNA and protein levels and IL-6 and IL-8 secretion from SZ95 sebocytes. Co-incubation of SZ95 sebocytes with MALP-2/FPCA did not affect the MALP-2-induced SCD mRNA upregulation but reduced FADS2 mRNA levels and inhibited IL-8 secretion. CRH induced an early, low-level SCD and FADS2 upregulation and TLR-2 and IL-8 secretion. High intracellular CRH concentrations could be detected early after CRH treatment and persisted up to 24h. MALP-2 stimulated intracellular CRH levels. MALP-2 stimulates the inflammatory signalling in human sebocytes through SCD and FADS2 activation. Inhibition of FADS2 mRNA levels and IL-8 secretion through MALP-2/FCPA co-incubation and diminution of fatty acid unsaturation might lead to a reduction of pro-inflammatory sebaceous lipids. CRH upregulates inflammatory signalling via the SCD/FADS2 pathway, and MALP-2 selectively enhances CRH levels in human sebocytes.

Efficacy and safety of bempedoic acid alone or combining with other lipid-lowering therapies in hypercholesterolemic patients: a meta-analysis of randomized controlled trials

BackgroundBempedoic acid is a new drug that reduces cholesterol synthesis via inhibiting ATP citrate lyase. It remains unclear whether the combination of bempedoic acid and other lipid-lowering drugs is better than these drugs alone. This study systematically reviewed the efficacy and safety of bempedoic acid monotherapy or combination togethers in hypercholesterolemic patients.MethodsRandomized controlled trials were searched across Medline, Embase, Cochrane library, web of science, etc. The net change scores [least squares mean (LSM) percentage change] in LDL-C level were meta-analyzed using weighted mean difference. The reductions in other lipids including total cholesterol (TC), non-high-density lipoprotein cholesterol (non-HDL-C) and apolipoprotein (ApoB) and high sensitivity C reactive protein (hsCRP) were also assessed. Odds ratio (OR) of the incidence of adverse events (AEs) were calculated to evaluate the safety of bempedoic acid.ResultsA total of 13 trials (4858 participates) were included. Pooled data showed that the combination togethers resulted in greater reductions in LDL-C level than monotherapies (bempedoic acid + statin vs. statin: LSM difference (%), − 18.37, 95% CI, − 20.16 to − 16.57, I2 = 0; bempedoic acid + ezetimibe vs. ezetimibe: LSM difference (%), − 18.89, 95% CI, − 29.66 to − 8.13, I2 = 87%). But the difference in efficacy between bempedoic acid and ezetimibe was not obvious. Meta-regression analysis showed the treatment duration was a source of heterogeneity (adj R2 = 16.92, 95% CI, 0.04 to 0.72). Furthermore, the background therapy of statin before screening decreased the efficacy of bempedoic acid. In addition, bempedoic acid also resulted in a significant reduction in TC, non-HDL-C, ApoB and hsCRP level. The OR of muscle-related AEs by the combination of bempedoic acid and statin was 1.29 (95% CI, 1.00 to 1.67, I2 = 0) when compared with statin alone.ConclusionThis study showed the efficacy of combination togethers were similar but stronger than these drugs alone. Of note, a trend of high risk of muscle-related AEs by the combination of bempedoic acid and statin was observed, though it is not statistically significant, such risk is needed to be confirmed by more trials, because it is important for us to determine which is the better combinative administration for statin-intolerant patients.

Short-Term Tea Consumption Is Not Associated with a Reduction in Blood Lipids or Pressure: A Systematic Review and Meta-Analysis of Randomized Controlled Trials

A recent systematic review of epidemiological evidence suggests that higher amounts of tea intake are associated with lower risks of cardiovascular disease (CVD) incidence and mortality. Our study objective was to assess mechanisms by which tea consumption may influence CVD risks. A systematic review and meta-analysis was conducted to investigate the effects of green and/or black tea consumption (≥4 wk) on systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride (TG) in healthy populations and among at-risk adults (analyzed separately) with metabolic syndrome, prediabetes, and hypercholesterolemia. The Grading of Recommendations Assessment, Development and Evaluation approach was used to rate the strength of evidence (SoE). A total of 14 unique RCTs which randomly assigned 798 participants to either green tea, black tea, or placebo controls were included in our analyses. Intervention durations ranged from 4 to 24 wk (mean: 7.4 wk). Individual studies were judged as moderate to high quality based on risk of bias assessments. SoE was low to moderate owing to low sample sizes and insufficient power for most included studies to observe changes in the measured CVD biomarkers. Meta-analyses showed no significant effects of tea consumption on SBP, DBP, total cholesterol, LDL cholesterol, HDL cholesterol, and TG in healthy and at-risk adults (i.e., adults with obesity, prediabetes, borderline hypercholesterolemia, and metabolic syndrome). Short-term (4-24 wk) tea consumption does not appear to significantly affect blood pressure or lipids in healthy or at-risk adults, although the evidence is limited by insufficient power to detect changes in these CVD biomarkers. High-quality RCTs with longer durations and sufficient sample sizes are needed to fully elucidate the effects of tea. This systematic review was registered at www.crd.york.ac.uk/prospero/ as CRD42020134513.

423 WS Young Scholar Talk MS II: Heat stress-induced deficits in growth, metabolic efficiency, and cardiovascular function coincided with chronic systemic inflammation and hypercatecholaminemia in ractopamine-supplemented feedlot lambs

Abstract Our 1st study evaluated the effects of □-adrenergic agonist (ractopamine) supplementation on growth, health, and wellbeing in heat-stressed feedlot wethers. Chronic heat stress impaired growth, metabolism, and wellbeing even when the impact of reduced feed intake was eliminated by pair-feeding. We observed systemic inflammation and hypercatecholaminemia that were likely mediators of these deficits. Moreover, ractopamine did not diminish any wellbeing indicators and improved muscle growth without worsening the effects of heat stress. Our 2nd study assessed the effects of pubertal cyclicity (weaning to 1st breeding) in cows on myoblast-mediated muscle growth, metabolism, and growth efficiency of their offspring. Maternal fertility and postnatal growth efficiency of offspring are imperative to successful beef cattle production. A maternal condition was previously identified in which cows exhibited irregular cyclicity patterns or were non-cyclic between weaning and 1st breeding. This condition is thought to be associated with high concentrations of androstenedione in their follicular fluid, which causes cows to be sub-fertile but also wean heavier calves when they do become pregnant. We hypothesized that calves from cows exhibiting irregular pubertal cyclicity would have enhanced growth efficiency compared to calves from cows with normal pubertal cyclicity. To test this hypothesis, we evaluated myoblast function, growth, and metabolism pre-weaning, as well as growth efficiency in the feedlot and carcass characteristics at harvest in calves from cows that were previously identified as having typical, start-stop, or non-cycling pubertal cyclicity patterns. Calves from irregular cycling cows had reductions in insulin sensitivity, plasma proteins and lipids paired with increased myoblast function and reduced myoblast glycolytic capacity. Data independently and combined suggest calves from cows with irregular cyclicity from weaning to 1st breeding have chronic inflammation. While the specific mechanism of inflammation is unknown, further research may allow mediators of abnormal cyclicity and offspring inflammation-induced dysfunction to be elucidated.

Effect of Resistant Starch (RS) Rich Sorghum Food Consumption on Lipids and Glucose Levels of Diabetic Subjects

Resistant starches are an important class, which gives benefits of fiber without affecting the sensory characteristics. Few studies reported the beneficial effect of resistant starch supplementation in reduction of lipid and glucose levels in diabetic subjects. In the present study we investigated the Resistant Starch (RS) rich millet food on lipid and glucose levels in diabetic subjects. Supplementation of 65 g of RS rich rawa (broken sorghum) for 90 days, significantly reduced Body Mass Index (BMI), Fasting Glucose (FG), TC (Total Cholesterol) and LDL-C (Low Density Lipo protein) (p,0.05) in diabetic subjects (n=15), while a non-significant reduction was found in HbA1c, eAG (estimated Average Glucose), TG, HDL-C and VLDL-C. The study indicated that regular consumption of RS rich foods might be beneficial for the diabetic population. Studies in larger population further strengthen the present understanding.

Performance evaluation of ultra violet assisted vertical re-circulating depuration system on microbial, heavy metal reduction and composition of black clam (Villorita cyprinoides)

A cost-effective, portable, ultra violet assisted vertical re-circulatory depuration system (UVDC) was developed and its effect on microbiological, heavy metal and proximate composition of Black clam (Villorita cyprinoides) was investigated. The process of depuration was found to be effective in reduction of total viable count by 1.8 log cfu g-1. Escherichia coli were within the permissible limit and both Vibrio cholera and V. parahaemolyticus were absent. The concentrations of heavy metals (Cd, Pb and Cu) were found to decrease by 40.44%, 96.51% and 34.50% respectively at the end of 48 h of depuration in UVDC. Significant reduction in protein, lipid, ash and glycogen contents were observed after the depuration process. The portability and easy to assemble nature of the system further improves its applicability as an onsite depuration unit which could be effectively utilized by the small-scale clam fishers/farmers depending on polluted waters for enhancing their income and livelihood.

Transient ischemic attack presenting as recurrent migratory numbness by seconds: a rare case confirmed by transcranial Doppler micro-emboli monitoring

BackgroundTransient ischemic attack (TIA) is a brief episode of cerebral ischemia. However, if a symptom is not presented as drop attack or hemiplegia, and alarming to the patient and the physician, how short of a symptom duration would raise the concern of a physician for TIA? It will be more complicated if the location of the neurological deficit is vagrant. This report highlights a rare TIA case which presented a very short duration of migratory patchy distribution numbness.Case presentationA middle-aged gentleman was presented with recurrent patchy distribution numbness on the right side of the body for 2 months, with the episode lasting as short as about 10 s. The location of the numbness was erratic and migratory. Magnetic resonance angiography (MRA) revealed mild stenosis on the left middle cerebral artery (MCA). Transcranial Doppler (TCD) micro-emboli monitoring detected positive micro-emboli signals (MES), leading to the confirmation of a TIA diagnosis. After a standard dual antiplatelet treatment combined with enhanced lipid reduction therapy with statins, MES disappeared on dynamic TCD emboli monitoring, and no more episodes of TIA have been noticed on the follow-ups.ConclusionTIA caused by micro-emboli can display as recurrent migratory neurological deficit within seconds. TCD micro-emboli monitoring is very helpful to differentiate this situation from TIA mimics with follow-ups, as well as to locate unstable plague.

Abstract 13405: Genetic Deletion of Stimulator of Interferon Genes Attenuates Atherogenesis in Apolipoprotein E-deficient Mice

Introduction: Recent studies show the contribution of innate immunity system to the pathogenesis of inflammatory diseases, including atherosclerosis. Stimulation of interferon genes (STING), originally known as a cytosolic DNA sensor, recognizes cytosolic DNA fragments, activating innate immunity. Here, we investigated whether STING contributes to the development of vascular inflammation and atherogenesis in apolipoprotein E-deficient (Apoe –/– ) mice. Methods and Results: The expression of STING increased in the atherosclerotic aorta in both gene and protein expression levels. STING-deficient Apoe –/– (STING –/– Apoe –/– ) mice reduced atherosclerotic lesions in the aortic arch ( P &lt;0.05), along with the reduction of lipid and macrophage accumulation in atherosclerotic plaques ( P &lt;0.05, respectively), and inflammatory molecule expression in the aorta compared with those in Apoe –/– mice after 20 weeks on a western-type diet. Also, pharmacologic blockade of STING in Apoe –/– mice for 12 weeks treatment attenuated atherogenesis in the aortic arch ( P &lt;0.05), reduced the accumulation of lipid in atherosclerotic plaques ( P &lt;0.05) with no alteration of metabolic parameters. Restoration of STING in bone marrow in STING –/– Apoe –/– mice promoted atherogenesis ( P &lt;0.05), lipid deposition ( P &lt;0.05), and vascular inflammation. cGAMP, a specific STING agonist, or mitochondrial DNA extracted from macrophages promoted expression of inflammatory molecules more effectively in Apoe -/- macrophages than in STING –/– Apoe –/– macrophages, while C-176, a specific STING inhibitor, attenuated these inflammatory responses. The results of western blotting showed the involvement of NF-κB and IRF-3 signaling in STING-associated vascular inflammation and macrophage activation. Furthermore, in humans, STING expression was confirmed in atherosclerotic lesions in the carotid artery. Conclusion: STING signaling activates macrophages, promotes vascular inflammation and atherosclerosis in Apoe -/- mice. Our results suggest that STING may serve as a potential therapeutic target for atherosclerosis.

A Comparison of Physical, Chemical, Biological and Combined Treatments for Detoxification of Free Gossypol in Crushed Whole Cottonseed

Oilseed plants such as cotton (Gossypium sp.) generate abundant biomass residues which contain significant levels of edible oil, crude proteins and other desirable biomolecules for the animal nutrition industry. The application of cottonseed cake in animal feed, a by-product of the cotton industry, is limited due to the natural presence of toxic free gossypol (FG), wherein efficient and cost-effective methods for FG detoxification are necessary. Herein, pretreatment methods for reducing FG in crushed whole cottonseed (CWCS) were compared, with residual FG quantified using a sensitive Ultra High-Performance Liquid Chromatography method for detection at trace levels in cottonseed materials. Physical treatment by autoclaving resulted in up to 96% detoxification of FG, without reduction in crude protein (CP) content. Chemical treatment with 1% and 2% Ca(OH)2 eliminated FG to as low as 0.04%, although a reduction in CP content was observed. Similarly, native fermentation, whilst reducing FG content by 99.66% after 6 days incubation, also reduced CP content. In combined physical and biological solid-state fermentation (SSF), basidiomycete fungi Ganoderma lucidum CC351, Panus lecomtei CC40, Pleurotus ostreatus CC389, Pleurotus sapidus CC28 and Pycnoporus sanguineus CC400 all degraded FG in autoclaved CWCS to trace levels often lower than obtained by individual treatments. A reduction in total lipids and increase in CP were also observed, improving nutritional quality. The most efficient fungi, P. ostreatus CC389 and P. lecomtei CC40, secreted considerable laccase and manganese peroxidase enzymes during SSF, potentially involved in FG detoxification. Cost effective, non-polluting, value-adding approaches for FG detoxification offer potential in animal feed industries.

Using coronary artery calcium to guide cardiovascular risk reduction: insights from the CAUGHT-CAD trial

Abstract Introduction International guidelines now recognise the value of CT coronary artery calcium scoring (CAC) to guide statin prescribing decisions. However, evidence is conflicting on CAC's potential to reinforce clinical advice, promote adherence and facilitating positive health behavioural change. Purpose We sought to assess the impact of a CAC-guided statin treatment protocol on cardiovascular risk at 12 months. Methods The CAUGHT-CAD trial recruited 455 asymptomatic, statin-naïve individuals at intermediate risk CVD risk with a family history of premature coronary artery disease (CAD) and CAC scores between 1 and 400. Participants were randomised to visualisation of their CAC or remain blinded. Both groups were provided standard risk factor advice. We calculated the 10-year SCORE predicted risk based on participant's baseline and 12 month risk factor profile. Paired t test and regression analysis was used to assess change in predicted risk both within and between treatment group. Results CAC-visualisation and statin treatment resulted in a 0.45% absolute and 21% relative risk reduction in predicted SCORE risk at 1 year. The CAC exposure group demonstrated 2.78 mmHg reduction in systolic blood pressure compared to the blinded group (p&amp;lt;0.01); accounting for 25% of the predicted absolute risk reduction despite both groups reporting the same level of dietary and physical activity goals achieved. 9 blinded participants who were commenced on statins off-protocol showed attenuated reductions in lipids compared to the CAC exposed group (total cholesterol: m-0.5 vs. −1.5 mmol/L, p&amp;lt;0.01; LDL −0.6 mmol/L vs. −1.4 mmol/L, p&amp;lt;0.01) suggesting poorer adherence. Conclusion CAC visualisation can augment primary CVD prevention by improving systolic blood pressure and absolute reduction in lipids with statin treatment. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): NHMRC project grant

Network Pharmacology Combined with Transcriptional Analysis to Unveil the Biological Basis of Astaxanthin in Reducing the Oxidative Stress Induced by Diabetes Mellitus.

PurposeAstaxanthin (Ast) has been reported to reduce oxidative stress induced by diabetes mellitus (DM). The aim of this research was to give a systematic overview of the biological basis for this process.MethodsAst-targeted proteins were collected from the BATMAN database, Comparative Toxicogenomics Database, and STITCH database. Putative DM-related protein targets were collected from the GeneCards database. A DM-rat model was then built with streptozotocin (STZ) combined with a high-sugar, high-fat diet for 30 days. Total cholesterol (TC), triglycerides (TGs), and insulin levels were examined using whole tail-vein blood from overnight-fasted rats. SOD, GSH, and MDA activy was detected in liver tissue (p<0.05). In addition, we used RNA-sequencing analysis to detect gene-transcription level in liver tissue of rats and GO biological process analysis to show all the log2FC≥2 genes in the Ast-fed DM rats compared with the DM group using the STRING database. Ast-intersecting targets were collected with Venn analysis. Docking analysis between Ast and targeted proteins was down with the SwissDock server. Ast targets–pathway networks were built using Cytoscape 3.7.2 software.ResultsA total of 120 Ast-targeted proteins and 13,784 DM-related targets were collected. Ast functioned in reducing TC, TG, and MDA levels, promoting SOD activity and GSH expression, and alleviating islet-cell injury in Ast-fed DM rats compared with DM control rats. Furthermore, genes involved in MAPK, TNF, AMPK, and FOXO signaling pathways were differently expressed in Ast-treated DM rats compared with DM rats. The differentially expressed genes were enriched in euchromatin, thyroid cancer, and metaphase-plate congression. Three Ast-intersecting targets — Col5A1, Nqo1, and Notch2 — were then identified. We found possible binding patterns of Ast with Nqo1 and Notch2, respectively. Ast targets–pathway networks were finally built to show a systematic overview of how Ast works in multiple pathways to reduce oxidative stress. Taken together, Ast is predicted to target Col5A1, Nqo1, and Notch2 to form a network of systemic pharmacological effects to: 1) promote insulin-releasing balance and relieve insulin resistance, 2) reduce testicular cell apoptosis, and 3) maintain normal size in marginal-zone B cells and inhibit autoimmune DM, all of which contribute to the balance of lipid metabolism and reduction of oxidative stress in DM patients.ConclusionAst functions in reducing oxidative stress in DM rats by regulating a variety of targets to form a comprehensive antioxidative network.

Metabolic and hormonal side effects of mitotane treatment for adrenocortical carcinoma: A retrospective study in 50 Danish patients.

Mitotane is used in the treatment of adrenocortical carcinoma (ACC). Metabolic and hormonal side effects of mitotane, the effect of subsequent treatment with statins and hormones and the effects of discontinuation of mitotane were assessed. Fifty patients were included. Lipid profiles, thyroid hormones, sex hormones and adrenal function from first year of mitotane treatment and after cessation were evaluated. After 6months of mitotane treatment total cholesterol increased from (median) 5.1 (IQR 4.3 to 5.8) to 7.4 (6.2-9.0)mmol/L, p<.001. LDL, HDL and triglyceride also increased, all p≤.03. Three months of treatment with statins decreased total and LDL-cholesterol, and cessation of mitotane led to further reduction in lipids. Plasma thyroxine decreased from 90 (78-111) to 57 (47-63)nmol/L and free thyroxine from 16.0 (13.0-18.3) to 11.7 (10.5-12.6)pmol/L on mitotane, both p<.001, while TSH remained unchanged. Treatment with thyroxin significantly increased plasma thyroxine and free thyroxine and decreased TSH. Cessation of mitotane increased total T4 (p<.001). Mitotane increased plasma SHBG from 36 (22-51) to 189 (85-259)nmol/L and LH from 4.6 (1.6-8.1) to 20.0 (10.0-34.9)IU/L, both p<.001. In males the changes were accompanied by an increase in testosterone from 9.8 (7.2-14.5) to 27.0 (15.3-34.8)nmol/L, p<.03. Fifteen of 24 tested patients regained normal adrenal function 6 (3-16)months after cessation of mitotane. Mitotane treatment exerts multiple severe side effects involving both the metabolic and endocrine systems that may require treatment, but the effect appears to be partially reversible.

The effect of dietary nitrate and vitamin C on endothelial function, oxidative stress and blood lipids in untreated hypercholesterolemic subjects: A randomized double-blind crossover study

Vitamin C may enhance nitric oxide (NO) production through stepwise reduction of dietary nitrate (NO3) to nitrite (NO2) to NO. The combined effect of vitamin C and NO3 supplementation is relatively unexplored in untreated hypercholesterolemia. We aimed to examine whether co-administration of vitamin C and nitrate for 4-weeks would improve endothelial function (primary outcome), plasma NO metabolites, oxidative stress, and blood lipids (secondary outcomes). Subjects 50-70 years of age with low density lipoprotein (LDL)>130mg/dL and RHI ≤2 were enrolled in this randomized double-blind crossover study. Subjects were assigned to two 4-week supplementation treatments starting with 70ml of concentrated beetroot juice (CBJ) with 1000mg of vitamin C (NC) or CBJ with matched placebo (N), then switched to alternate treatment following 2-week washout. The change in reactive hyperemia index (RHI), sum of plasma NO metabolites (NO2+NO3 (NOx)), oxidized LDL (oxLDL), and serum lipids were assessed at baseline and at 4-weeks of each treatment period. Eighteen subjects (11M:7F) completed all study visits. No significant treatment differences were observed in RHI change (N: 0.21±0.12; NC: 0.20±0.17; p=0.99). Secondary analysis revealed that a subgroup of NC subjects who started with a baseline RHI of <1.67 (threshold value for ED) had greater improvements in RHI compared to subjects with RHI >1.67 (1.23±0.15 to 1.96±0.19; n=8 vs. 1.75±0.11 to 1.43±0.10; n=8; p=0.02). Compared to N, NC experienced a significant increase in plasma NOx (N: 94.2±15.5μmol/L; NC: 128.7±29.1μmol/L; p=0.01). Although there was no significant difference in oxLDL change between treatments (N:-1.08±9.8 U/L; NC:-6.07±9.14 U/L; p=0.19), NC elicited significant reductions in LDL (N: 2.2±2; NC:-10.7±23; p=0.049), triglycerides (N: 14.6±43; NC:-43.7±45; p=0.03), and no change in serum high density lipoprotein. Within treatment group comparisons showed that only NC reduced oxLDL significantly from baseline to 4 weeks (p=0.01). No between intervention differences were observed in RHI. RHI only improved in NC subjects with ED at intervention baseline. Four weeks of NC enriched the NO pool and promoted reduction of blood lipids and oxidative stress in subjects with hypercholesterolemia. These preliminary findings highlight a supplementation strategy that may reduce the progression of atherosclerotic disease and deserves further attention in studies using flow mediated dilation methods. www.clinicaltrials.gov (NCT04283630).

The Tmem43-S358l Mutation Affects Cardiac and Small Intestine Homeostasis Contributing to Arvc5 Pathogenesis in Vivo

Objectives: This study sought to delineate clinical phenotype and investigate pathophysiological mechanisms of arrhythmogenic right ventricular cardiomyopathy (ARVC5) in vivo.Background: Mutation p.S358L in TMEM43/LUMA causes ARVC5, a fully penetrant gender- and exercise-dependent disease. Exact pathophysiological mechanisms of ARVC5 remain incomplete and controversial. Methods: Knock-in Tmem43S358L mice were generated using conventional targeting. Assessment of cardiac and intestinal phenotypes, transcriptome and protein analysis were performed. Results: Onset of systolic dysfunction and rhythm asymmetry was apparent in 3-month-old homozygote mutants. Heterozygote 6-month-old mutants displayed systolic dysfunction as well. Biventricular myocardial fibro-fatty infiltration and subcellular abnormalities were found in both mutants, while acute stress caused ventricular tachycardia and death in homozygotes. Microarray revealed that WNT-b-catenin and PPARG signaling-associated genes were differentially expressed between LV and RV. Tmem43S358L mutants displayed significantly reduced PPARG activities and downregulation of PPARG-downstream metabolic genes in the heart. Expression of b-catenin were significantly reduced, while JUP was translocated into nuclei of mutant cardiomyocytes. Conversely in small intestine, b-catenin and PPARG-associated genes responsible for cholesterol, bile acid, and lipid transport were upregulated in Tmem43S358L mutants resulting in elevated lipids absorption, hyperlipidemia, and reciprocal fecal lipids reduction. These abnormalities were associated with elongated villi, fatty infiltrations, and gut epithelial proliferation markers in small intestine. Conclusions: We defined novel distinct effects of the TMEM43-S358L mutation on small intestine homeostasis in addition to cardiac damage. Tmem43-S358L perturbs PPARG and WNT-b-catenin signaling in the heart as well as in intestine, elevating lipids absorption from gut lumen whereby triggering ACM pathophysiology.