The Beneficial Effects of Triterpenic Acid and Acteoside in an In Vitro Model of Nonalcoholic Steatohepatitis (NASH)

Triterpenic acid (TA) and acteoside (ACT), the major components of APPLIVER and ACTEOS, respectively, have been reported to exert hepatoprotective effects, but the molecular mechanisms remain elusive, particularly in the NAFLD/NASH context. We assessed their effects in our well-established in vitro model resembling the pathophysiological mechanisms involved in NASH. Human hepatocytes and hepatic stellate cells were exposed to free fatty acids (FFA) alone or in combination with APPLIVER and ACTEOS as a mono- or co-culture. Steatosis, inflammation, generation of reactive oxygen species (ROS), and collagen deposition were determined. ACTEOS reduced both the TNF-α and ROS production, and, most importantly, attenuated collagen deposition elicited by the excess of FFA in the co-culture model. APPLIVER also showed inhibition of both TNF-α production and collagen deposition caused by FFA accumulation. The compounds alone did not induce any cellular effects. The present study showed the efficacy of APPLIVER and ACTEOS on pathophysiological mechanisms related to NASH. These in vitro data suggest that these compounds deserve further investigation for possible use in NASH treatment.     

The Effects of Probiotics,Prebiotics and Synbiotics in Non-Alcoholic Fat Liver Disease (NAFLD) and Non-Alcoholic Steatohepatitis (NASH): A Systematic Review

Modifications in the microbiota caused by environmental and genetic reasons can unbalance the intestinal homeostasis, deregulating the host’s metabolism and immune system, intensifying the risk factors for the development and aggravation of non-alcoholic fat liver disease (NAFLD). The use of probiotics, prebiotics and synbiotics have been considered a potential and promising strategy to regulate the gut microbiota and produce beneficial effects in patients with liver conditions. For this reason, this review aimed to evaluate the effectiveness of probiotics, prebiotics, and symbiotics in patients with NAFLD and NASH. Pubmed, Embase, and Cochrane databases were consulted, and PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines were followed. The clinical trials used in this study demonstrated that gut microbiota interventions could improve a wide range of markers of inflammation, glycemia, insulin resistance, dyslipidemia, obesity, liver injury (decrease of hepatic enzymes and steatosis and fibrosis). Although microbiota modulators do not play a healing role, they can work as an important adjunct therapy in pathological processes involving NAFLD and its spectrums, either by improving the intestinal barrier or by preventing the formation of toxic metabolites for the liver or by acting on the immune system.     

Liver Bile Acid Changes in Mouse Models of Alzheimer’s Disease

Alzheimer’s disease (AD) is a neurodegenerative disease characterized by progressive cognitive impairment. It is hypothesized to develop due to the dysfunction of two major proteins, amyloid-β (Aβ) and microtubule-associated protein, tau. Evidence supports the involvement of cholesterol changes in both the generation and deposition of Aβ. This study was performed to better understand the role of liver cholesterol and bile acid metabolism in the pathophysiology of AD. We used male and female wild-type control (C57BL/6J) mice to compare to two well-characterized amyloidosis models of AD, APP/PS1, and App^NL-G-F. Both conjugated and unconjugated primary and secondary bile acids were quantified using UPLC-MS/MS from livers of control and AD mice. We also measured cholesterol and its metabolites and identified changes in levels of proteins associated with bile acid synthesis and signaling. We observed sex differences in liver cholesterol levels accompanied by differences in levels of synthesis intermediates and conjugated and unconjugated liver primary bile acids in both APP/PS1 and App^NL-G-F mice when compared to controls. Our data revealed fundamental deficiencies in cholesterol metabolism and bile acid synthesis in the livers of two different AD mouse lines. These findings strengthen the involvement of liver metabolism in the pathophysiology of AD.     

Lessons on Drug Development: A Literature Review of Challenges Faced in Nonalcoholic Fatty Liver Disease (NAFLD) Clinical Trials

NAFLD is the most common chronic liver disease worldwide, occurring in both obese and lean patients. It can lead to life-threatening liver diseases and nonhepatic complications, such as cirrhosis and cardiovascular diseases, that burden public health and the health care system. Current care is weight loss through diet and exercise, which is a challenging goal to achieve. However, there are no FDA-approved pharmacotherapies for NAFLD. This review thoroughly examines the clinical trial findings from 22 drugs (Phase 2 and above) and evaluates the future direction that trials should take for further drug development. These trialed drugs can broadly be categorized into five groups—hypoglycemic, lipid-lowering, bile-pathway, anti-inflammatory, and others, which include nutraceuticals. The multitude of challenges faced in these yet-to-be-approved NAFLD drug trials provided insight into a few areas of improvement worth considering. These include drug repurposing, combinations, noninvasive outcomes, standardization, adverse event alleviation, and the need for precision medicine with more extensive consideration of NAFLD heterogenicity in drug trials. Understandably, every evolution of the drug development landscape lies with its own set of challenges. However, this paper believes in the importance of always learning from lessons of the past, with each potential improvement pushing clinical trials an additional step forward toward discovering appropriate drugs for effective NAFLD management.     

Multiple Hits,Including Oxidative Stress,as Pathogenesis and Treatment Target in Non-Alcoholic Steatohepatitis (NASH)

Multiple parallel hits, including genetic differences, insulin resistance and intestinal microbiota, account for the progression of non-alcoholic steatohepatitis (NASH). Multiple hits induce adipokine secretion, endoplasmic reticulum (ER) and oxidative stress at the cellular level that subsequently induce hepatic steatosis, inflammation and fibrosis, among which oxidative stress is considered a key contributor to progression from simple fatty liver to NASH. Although several clinical trials have shown that anti-oxidative therapy can effectively control hepatitis activities in the short term, the long-term effect remains obscure. Several trials of long-term anti-oxidant protocols aimed at treating cerebrovascular diseases or cancer development have failed to produce a benefit. This might be explained by the non-selective anti-oxidative properties of these drugs. Molecular hydrogen is an effective antioxidant that reduces only cytotoxic reactive oxygen species (ROS) and several diseases associated with oxidative stress are sensitive to hydrogen. The progress of NASH to hepatocellular carcinoma can be controlled using hydrogen-rich water. Thus, targeting mitochondrial oxidative stress might be a good candidate for NASH treatment. Long term clinical intervention is needed to control this complex lifestyle-related disease.     

Plasma Aldo-Keto Reductase Family 1 Member B10 as a Biomarker Performs Well in the Diagnosis of Nonalcoholic Steatohepatitis and Fibrosis

We found several blood biomarkers through computational secretome analyses, including aldo-keto reductase family 1 member B10 (AKR1B10), which reflected the progression of nonalcoholic fatty liver disease (NAFLD). After confirming that hepatic AKR1B10 reflected the progression of NAFLD in a subgroup with NAFLD, we evaluated the diagnostic accuracy of plasma AKR1B10 and other biomarkers for the diagnosis of nonalcoholic steatohepatitis (NASH) and fibrosis in replication cohort. We enrolled healthy control subjects and patients with biopsy-proven NAFLD (n = 102) and evaluated the performance of various diagnostic markers. Plasma AKR1B10 performed well in the diagnosis of NASH with an area under the receiver operating characteristic (AUROC) curve of 0.834 and a cutoff value of 1078.2 pg/mL, as well as advanced fibrosis (AUROC curve value of 0.914 and cutoff level 1078.2 pg/mL), with further improvement in combination with C3. When we monitored a subgroup of obese patients who underwent bariatric surgery (n = 35), plasma AKR1B10 decreased dramatically, and 40.0% of patients with NASH at baseline showed a decrease in plasma AKR1B10 levels to below the cutoff level after the surgery. In an independent validation study, we proved that plasma AKR1B10 was a specific biomarker of NAFLD progression across varying degrees of renal dysfunction. Despite perfect correlation between plasma and serum levels of AKR1B10 in paired sample analysis, its serum level was 1.4-fold higher than that in plasma. Plasma AKR1B10 alone and in combination with C3 could be a useful noninvasive biomarker for the diagnosis of NASH and hepatic fibrosis.     

Serum cholesterol,triglycerides and total lipid fatty acids of rats in response to okra(hibiscus escuientus) seed oil

Tender pods of okra are commonly consumed vegetables in India. Okra seed kernel, like soybean, is a rich source of protein and fat. Its fat, with its appreciable linoleic acid content (>42%), prompted us to look into its metabolic utility in comparison with commonly consumed groundnut oil. Serum lipid profiles, with respect to cholesterol, triglycerides and total lipid fatty acids were determined in rats receiving okra seed oil at a level of 10% in the casein based diet which was adequate with respect to vitamins, minerals, etc. The control group received a casein based diet in which groundnut oil was the source of fat. Serum lipid profiles in this group were similarly monitored. The feeding trial was carried out for a period of 90 days. Results showed that serum cholesterol content of rats receiving okra seed oil was significantly lower compared to those consuming groundnut oil. A decreasing trend in total lipids as well as triglycerides was also evident in animals fed okra seed oil. Serum fatty acid profiles showed a relatively higher proportion of long chain and polyunsaturated fatty acids in this group as compared to the group receiving groundnut oil. These results indicate that okra seed oil consumption has a potential hypocholesterolemic effect. To whom correspondence to be addressed. ¹Part of this work was presented at 45th Annual Meeting of Oil Technologists Association of India, New Delhi-Feb. 9–10, 1990.     

Acute Fasting Induces Expression of Acylglycerophosphate Acyltransferase (AGPAT) Enzymes in Murine Liver,Heart, and Brain

During fasting, cells increase uptake of non-esterified fatty acids (NEFA) and esterify excess into phosphatidic acid (PtdOH), the common precursor of both triacylglycerols and phospholipids, using acylglycerophosphate acyltransferases/lysophosphatidic acid acyltransferases (AGPAT/LPAAT). Knowledge of the regulation of AGPAT enzymes is important for understanding fasting adaptations. Total RNA was isolated from liver, heart, and whole brain tissue of C57BL/6J mice fed ad libitum, or fasted for 16 h. Following fasting, induction of Agpat2, 3, 4, and 5 was observed in the liver, Agpat2 and 3 in heart tissue, and Agpat1, 2, and 3 in whole brain tissue. As a result, the relative abundance profile of the individual homologues within specific tissues was found to be significantly altered depending on the nutritive state of the animal. These data demonstrate tissue-specific effects of fasting on the regulation of different Agpat that are implicated in supporting unique downstream glycerolipid synthesis pathways.     

Development of Sesamol Carbamate-L-Phenylalanine Prodrug Targeting L-Type Amino Acid Transporter1 (LAT1) as a Potential Antiproliferative Agent against Melanoma

Sesamol is a compound reported to have anti-melanogenesis and anti-melanoma actions. Sesamol, however, has low intracellular drug concentration and fast excretion, which can limit its benefits in the clinic. To overcome this drawback and increase intracellular delivery of sesamol into the target melanoma, research has focused on L-type amino acid transporter 1 (LAT1)-mediated prodrug delivery into melanoma cells. The sesamol prodrug was designed by conjugating sesamol with L-phenylalanine at the para position with a carbamate bond. LAT1 targeting was evaluated vis-à-vis a competitive [¹⁴C]-leucine uptake inhibition. The sesamol prodrug has a higher [¹⁴C]-leucine uptake inhibition than sesamol in human LAT1-transfected HEK293 cells. Moreover, the sesamol prodrug was taken up by LAT1-mediated transport into SK-MEL-2 cells more effectively than sesamol. The sesamol prodrug underwent complete hydrolysis, releasing the active sesamol at 72 h, which significantly exerted its cytotoxicity (IC₅₀ of 29.3 µM) against SK-MEL-cells more than sesamol alone. Taken together, the strategy for LAT1-mediated prodrug delivery has utility for the selective uptake of sesamol, thereby increasing its intracellular concentration and antiproliferation activity, targeting melanoma SK-MEL-2 cells that overexpress the LAT1 protein. The sesamol prodrug thus warrants further evaluation in an in vivo model.     

Electrochemical oxidation of adsorbed alkenoic acids as a function of chain length at Pt(111) electrodes. Studies by cyclic voltammetry,EELS and Auger spectroscopy

Studies of the electrochemical oxidation of a series of straight-chain terminal alkenoic acids adsorbed at a Pt(111) electrode surface are reported. Compounds adsorbed were: propenoic acid (acrylic acid, PPA); 3-butenoic acid (vinylacetic acid, 3BTA); 4-pentenoic acid (allylacetic acid, 4PTA); 6-heptenoic acid (6HPA); and 10-undecenoic acid (10UDA). Vibrational spectra of adsorbed layers were obtained by use of electron energy-loss spectroscopy (EELS). Molecular packing densities were measured by use of Auger spectroscopy. Electrochemical oxidation of each adsorbed layer was explored by means of cyclic voltammetry in aqueous inert electrolyte (KF/HF). As the analogous aliphatic acids are not chemisorbed at Pt under the same conditions, the alkenoic acids evidently adsorb at Pt(111) predominantly through the C=C double bond. Molecular packing densities indicate that the carboxylic acid moiety is in contact with the Pt surface only in the case of PPA. EELS spectra also indicate that the carboxylate groups (other than in PPA) are present as pendants. The carboxylic acid O-H stretching bands of most of the adsorbed acids are red-shifted and broadened, evidently due to extensive intermolecular hydrogen bonding; the exceptions are PPA, for which the interaction is primarily with the Pt surface, and 3BTA, for which intermolecular interaction between the carboxylic acid pendants is apparently prevented by steric considerations. The surface-attached carboxylic acid moieties react with KOH solution, leading to retention of K⁺ ions, detected by Auger spectroscopy, and to changes in the vibrational spectra indicative of carboxylate anions; reactivity toward KOH decreases with chain length. Adsorbed alkenoic acids at Pt(111) surfaces are stable in water and in vacuum. Oxidation of the adsorbed short-chain acids PPA and 3BTA proceeds to completion, forming CO₂ as the principal product. Oxidation of the adsorbed long-chain acids converts the C=C moiety to 2CO₂, and transforms the remainder of the molecule to an unadsorbed diacid (likely possibilities are malonic acid from 4PTA; glutaric acid from 6HPA; and heptane-1,7-dioic acid from 10UDA).     

Identification of Activated Protein Kinase Cα (PKCα) in the Urine of Orthotopic Bladder Cancer Xenograft Model as a Potential Biomarker for the Diagnosis of Bladder Cancer

Bladder cancer has a high recurrence rate; therefore, frequent and effective monitoring is essential for disease management. Cystoscopy is considered the gold standard for the diagnosis and continuous monitoring of bladder cancer. However, cystoscopy is invasive and relatively expensive. Thus, there is a need for non-invasive, relatively inexpensive urinary biomarker-based diagnoses of bladder cancer. This study aimed to investigate the presence of activated protein kinase Cα (PKCα) in urine samples and the possibility of PKCα as a urinary biomarker for bladder cancer diagnosis. Activated PKCα was found to be present at higher levels in bladder cancer tissues than in normal bladder tissues. Furthermore, high levels of activated PKCα were observed in urine samples collected from orthotopic xenograft mice carrying human bladder cancer cells compared to urine samples from normal mice. These results suggest that activated PKCα can be used as a urinary biomarker to diagnose bladder cancer. To the best of our knowledge, this is the first report describing the presence of activated PKCα in the urine of orthotopic xenograft mice.     

Investigation of the Pentathiepin Functionality as an Inhibitor of Feline Immunodeficiency Virus (FIV) via a Potential Zinc Ejection Mechanism,as a Model for HIV Infection

A small diverse library of pentathiepin derivatives were prepared to evaluate their efficacy against the nucleocapsid protein function of the feline immunodeficiency virus (FIV) as a model for HIV, using an in vitro cell culture approach. This study led to the development of nanomolar active compounds with low toxicity.     

Utilization of 2‐(4‐Nitrophenylsulfonyl)ethoxycarbonyl (Nsc) as a Substitute for 9H‐Fluoren‐9‐ylmethoxycarbonyl (Fmoc) in Liquid Phase Chemistry

2‐(4‐Nitrophenylsulfonyl)ethoxycarbonyl (Nsc) is a useful substitute for the Fmoc group. It is easily removed not only with secondary amines but with tris(aminoethyl)amine (TAEA) and with resin‐bound TAEA, thus allowing for a simplified work‐up: the side products of the deprotection are removed either by extraction with phosphate buffer or by filtration.     

腐植酸作为天然有机物(NOM)的主要类型对环糊精聚氨酯去除水中异味的影响

目前,水处理机构普遍认为,天然有机物会妨碍活性炭的吸附位点与异味化合物结合,进而导致对甲萘烷醇和2-甲基异茨醇的清除效率下降。腐植酸也被认为具有相同的作用。对于城市水处理系统来说,饮用水中异味化合物如甲萘烷醇和2-甲基异茨醇的清除至关重要。已证实,环糊精聚氨酯可清除水中有机污染物,但不会显著降低天然有机物的含量。我们希望确定在有天然有机物存在的条件下,该聚合物是否能够有选择地去除甲萘烷醇和2-甲基异茨醇。本研究以腐植酸为天然有机物代表,因腐植酸约占天然有机物的70%。研究表明,不同浓度腐植酸的存在不会影响甲萘烷醇和2-甲基异茨醇的清除,使用环糊精聚合物,90%的异味化合物被去除。同时,紫外分析表明只有少量的腐植酸(3%～20%)被去除。     

Development of a bioassay to test phloem sap samples from lettuce for resistance toNasonovia ribisnigri (homoptera,aphididae)

TheNr-gene-based resistance of lettuce to the aphidNasonovia ribisnigri (Mosley) has previously been shown to be located in the phloem. Since chemical analyses of the phloem sap had shown no differences between resistant and susceptible lines, a bioassay was developed in order to test samples from resistant and susceptible plants on aphid feeding. For this, whole-plant extracts, honeydew, and EDTA-collected phloem extracts were obtained, and a sensitive bioassay was developed using EDTA samples. The EDTA was removed, and samples were added to a simple sucrose solution or to a complex artificial diet and presented in a choice situation comparing extracts from resistant and susceptible plants. EDTA-collected phloem sap samples from susceptible plants were preferred to those from resistant plants. The resistance is probably based on a feeding deterrent activity of the phloem sap in the resistant plant.     

Valproic Acid as a Potential Inhibitor of Plasmodium falciparum Histone Deacetylase 1 (PfHDAC1): An in Silico Approach

A new Plasmodium falciparum histone deacetylase1 (PfHDAC1) homology model was built based on the highest sequence identity available template human histone deacetylase 2 structure. The generated model was carefully evaluated for stereochemical accuracy, folding correctness and overall structure quality. All evaluations were acceptable and consistent. Docking a group of hydroxamic acid histone deacetylase inhibitors and valproic acid has shown binding poses that agree well with inhibitor-bound histone deacetylase-solved structural interactions. Docking affinity dG scores were in agreement with available experimental binding affinities. Further, enzyme-ligand complex stability and reliability were investigated by running 5-nanosecond molecular dynamics simulations. Thorough analysis of the simulation trajectories has shown that enzyme-ligand complexes were stable during the simulation period. Interestingly, the calculated theoretical binding energies of the docked hydroxamic acid inhibitors have shown that the model can discriminate between strong and weaker inhibitors and agrees well with the experimental affinities reported in the literature. The model and the docking methodology can be used in screening virtual libraries for PfHDAC1 inhibitors, since the docking scores have ranked ligands in accordance with experimental binding affinities. Valproic acid calculated theoretical binding energy suggests that it may inhibit PfHDAC1.     

Development of poly(lactic acid) films with propolis as a source of active compounds: Biodegradability,physical, and functional properties

Active films (AFs) using poly(lactic acid) (PLA) as a polymeric matrix containing various propolis concentrations (5, 8.5, and 13%) as the active agent (AA) were developed using a casting method. The purpose was to determine the effects of the incorporation of AA on the physical properties of the films and to evaluate the antioxidant and antimicrobial activities. Tensile strength and elastic modulus of the AFs decreased relative to the control (PLA without AA). Introducing the active substances from propolis into the PLA also affected its thermal properties (glass transition). Adding AAs to the polymer generated more opacity with a green-yellowish color compared to the control. In addition, AFs exhibited reduced water vapor permeability as the AA concentration increased. Biodegradation assay showed that the AFs degraded faster than the control. AFs exhibited antioxidant activity, which was measured as the ability to scavenge free radicals (2,2-diphenyl-1-picrylhydrazyl and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate)), due to the presence of bioactive compounds (phenolics). Antimicrobial activity was evaluated against Escherichia coli and showed a reduction over 4-log cycles. Therefore, incorporation of propolis is a useful strategy for the development of active packaging with antioxidant and antimicrobial effects, which increase the shelf life of food products. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47090.     

Reducing Capacity,Chlorogenic Acid Content and Biological Activity in a Collection of Scarlet (Solanum aethiopicum) and Gboma (S. macrocarpon) Eggplants

Scarlet (Solanum aethiopicum) and gboma (S. macrocarpon) eggplants are important vegetables in Sub-Saharan Africa. Few studies have been made on these crops regarding the diversity of phenolic content and their biological activity. We have studied the reducing activity, the chlorogenic acid and other phenolic acid contents in a collection of 56 accessions of scarlet eggplant, including the four cultivated groups (Aculeatum, Gilo, Kumba, Shum) and the weedy intermediate S. aethiopicum-S. anguivi types, as well as in eight accessions of gboma eggplant, including the cultivated S. macrocarpon and its wild ancestor, S. dasyphyllum. A sample of the accessions evaluated in this collection has been tested for inhibition of nitric oxide (NO) using macrophage cell cultures. The results show that there is a great diversity in both crops for reducing activity, chlorogenic acid content and chlorogenic acid peak area (% of total phenolic acids). Heritability (H²) for these traits was intermediate to high in both crops. In all samples, chlorogenic acid was the major phenolic acid and accounted for more than 50% of the chromatogram peak area. Considerable differences were found among and within groups for these traits, but the greatest values for total phenolics and chlorogenic acid content were found in S. dasyphyllum. In most groups, reducing activity was positively correlated (with values of up to 0.904 in the Aculeatum group) with chlorogenic acid content. Inhibition of NO was greatest in samples having a high chlorogenic acid content. The results show that both crops are a relevant source of chlorogenic acid and other phenolic acids. The high diversity found also indicates that there are good prospects for breeding new scarlet and gboma eggplant cultivars with improved content in phenolics and bioactive properties.     

Forms of n‐3 (ALA,C18:3n‐3 or DHA,C22:6n‐3) Fatty Acids Affect Carcass Yield,Blood Lipids,Muscle n‐3 Fatty Acids and Liver Gene Expression in Lambs

The effects of supplementing diets with n‐3 alpha‐linolenic acid (ALA) and docosahexaenoic acid (DHA) on plasma metabolites, carcass yield, muscle n‐3 fatty acids and liver messenger RNA (mRNA) in lambs were investigated. Lambs (n = 120) were stratified to 12 groups based on body weight (35 ± 3.1 kg), and within groups randomly allocated to four dietary treatments: basal diet (BAS), BAS with 10.7 % flaxseed supplement (Flax), BAS with 1.8 % algae supplement (DHA), BAS with Flax and DHA (FlaxDHA). Lambs were fed for 56 days. Blood samples were collected on day 0 and day 56, and plasma analysed for insulin and lipids. Lambs were slaughtered, and carcass traits measured. At 30 min and 24 h, liver and muscle samples, respectively, were collected for determination of mRNA (FADS1, FADS2, CPT1A, ACOX1) and fatty acid composition. Lambs fed Flax had higher plasma triacylglycerol, body weight, body fat and carcass yield compared with the BAS group (P < 0.001). DHA supplementation increased carcass yield and muscle DHA while lowering plasma insulin compared with the BAS diet (P < 0.01). Flax treatment increased (P < 0.001) muscle ALA concentration, while DHA treatment increased (P < 0.001) muscle DHA concentration. Liver mRNA FADS2 was higher and CPT1A lower in the DHA group (P < 0.05). The FlaxDHA diet had additive effects, including higher FADS1 and ACOX1 mRNA than for the Flax or DHA diet. In summary, supplementation with ALA or DHA modulated plasma metabolites, muscle DHA, body fat and liver gene expression differently.