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Policosanols from various sources, such as sugar cane, rice bran, and insects, have been marketed to prevent dyslipidemia, diabetes, and hypertension by increasing the blood high-density lipoproteins cholesterol (HDL-C) levels. On the other hand, there has been no study on how each policosanol influences the quality of HDL particles and their functionality. Reconstituted high-density lipoproteins (rHDLs) with apolipoprotein (apo) A-I and each policosanol were synthesized using the sodium cholate dialysis method to compare the policosanols in lipoprotein metabolism. Each rHDL was compared regarding the particle size and shape, antioxidant activity, and anti-inflammatory activity in vitro and in zebrafish embryos. This study compared four policosanols including one policosanol from Cuba (Raydel® policosanol) and three policosanols from China (Xi’an Natural sugar cane, Xi’an Realin sugar cane, and Shaanxi rice bran). The synthesis of rHDLs with various policosanols (PCO) from Cuba or China using a molar ratio of 95:5:1:1 with palmitoyloleoyl phosphatidylcholine (POPC): free cholesterol (FC): apoA-I:PCO (wt:wt) showed that rHDL containing Cuban policosanol (rHDL-1) showed the largest particle size and the most distinct particle shape. The rHDL-1 showed a 23% larger particle diameter and increased apoA-I molecular weight with a 1.9 nm blue shift of the maximum wavelength fluorescence than rHDL alone (rHDL-0). Other rHDLs containing Chinese policosanols (rHDL-2, rHDL-3, and rHDL-4) showed similar particle sizes with an rHDL-0 and 1.1–1.3 nm blue shift of wavelength maximum fluorescence (WMF). Among all rHDLs, the rHDL-1 showed the strongest antioxidant ability to inhibit cupric ion-mediated LDL oxidation. The rHDL-1-treated LDL showed the most distinct band intensity and particle morphology compared with the other rHDLs. The rHDL-1 also exerted the highest anti-glycation activity to inhibit the fructose-mediated glycation of human HDL2 with the protection of apoA-I from proteolytic degradation. At the same time, other rHDLs showed a loss of anti-glycation activity with severe degradation. A microinjection of each rHDL alone showed that rHDL-1 had the highest survivability of approximately 85 ± 3%, with the fastest developmental speed and morphology. In contrast, rHDL-3 showed the lowest survivability, around 71 ± 5%, with the slowest developmental speed. A microinjection of carboxymethyllysine (CML), a pro-inflammatory advanced glycated end product, into zebrafish embryos resulted in severe embryo death of approximately 30 ± 3% and developmental defects with the slowest developmental speed. On the other hand, the phosphate buffered saline (PBS)-injected embryo showed 83 ± 3% survivability. A co-injection of CML and each rHDL into adult zebrafish showed that rHDL-1 (Cuban policosanol) induced the highest survivability, around 85 ± 3%, while rHDL-0 showed 67 ± 7% survivability. In addition, rHDL-2, rHDL-3, and rHDL-4 showed 67 ± 5%, 62 ± 37, and 71 ± 6% survivability, respectively, with a slower developmental speed and morphology. In conclusion, Cuban policosanol showed the strongest ability to form rHDLs with the most distinct morphology and the largest size. The rHDL-containing Cuban policosanol (rHDL-1) showed the strongest antioxidant ability against LDL oxidation, anti-glycation activity to protect apoA-I from degradation, and the highest anti-inflammatory activity to protect embryo death under the presence of CML.

Reconstituted high-density lipoproteins (rHDL) containing each policosanol from Cuba (Raydel®), China (Shaanxi Pioneer), and the United States (Lesstanol®) were synthesized to compare the physiological properties of policosanol depending on sources and origin countries. After synthesis with apolipoproteinA-I (apoA-I) into rHDL, all policosanols bound well with phospholipid and apoA-I to form discoidal rHDL. An rHDL containing Cuban policosanol (rHDL-1) showed the largest rHDL particle size of around 83 ± 3 nm, while rHDL containing Chinese policosanol (rHDL-2) or American policosanol (rHDL-3) showed smaller particles around 63 ± 3 nm and 60 ± 2 nm in diameter, respectively. The rHDL-1 showed the strongest anti-glycation activity to protect the apoA-I degradation of HDL from fructose-mediated glycation: approximately 2.7-times higher ability to suppress glycation and 1.4-times higher protection ability of apoA-I than that of rHDL-2 and rHDL-3. The rHDL-1 showed the highest antioxidant ability to inhibit cupric ion-mediated LDL oxidation in electromobility and the quantification of oxidized species. A microinjection of each rHDL into a zebrafish embryo in the presence of carboxymethyllysine (CML) showed that rHDL-1 displayed the strongest anti-oxidant activity with the highest embryo survivability, whereas rHDL-2 and rHDL-3 showed much weaker protection ability, similar to rHDL alone (rHDL-0). An intraperitoneal injection of CML (250 μg) into adult zebrafish caused acute death and hyperinflammation with an elevation of infiltration of neutrophils and IL-6 production in the liver. On the other hand, a co-injection of rHDL-1 resulted in the highest survivability and the strongest anti-inflammatory ability to suppress IL-6 production with an improvement of the blood lipid profile, such as elevation of HDL-C and lowering of the total cholesterol, LDL-cholesterol, and triglyceride. In conclusion, Cuban policosanol exhibited the most desirable properties for the in vitro synthesis of rHDL with the stabilization of apoA-I, the largest particle size, anti-glycation against fructation, and antioxidant activities to prevent LDL oxidation. Cuban policosanol in rHDL also exhibited the strongest in vivo antioxidant and anti-inflammatory activities with the highest survivability in zebrafish embryos and adults via the prevention of hyperinflammation in the presence of CML.

Main conclusionQuantification of cuticular waxes coupled with insect bioassays and feeding behavior analysis demonstrate that long-chain C32 fatty alcohol impacts host plant selection by aphids.Cuticular waxes constitute the first point of contact between plants and their environment, and it also protect plants from external stresses. However, the role of waxes in Sorghum bicolor (sorghum) against sugarcane aphid (Melanaphis sacchari), a relatively new and devastating pest of sorghum in the U.S., is not fully understood. In this study, we monitored sugarcane aphid behavior on two genotypes of young sorghum plants with different wax chemistry: a wild-type plant (bloom) with lower C32 alcohol cuticular wax, and a mutant plant (bloomless) with 1.6 times the amount of wax compared to wild-type plants. No-choice aphid bioassays revealed that sugarcane aphid reproduction did not vary between wild-type and the bloomless plants. Electrical Penetration Graph (EPG) monitoring indicated that the sugarcane aphids spent comparable amount of time feeding from the sieve elements of the wild-type and bloomless plants. However, aphids spent more time feeding on the xylem sap of the bloomless plants compared to the wild-type plants. Furthermore, aphid choice assays revealed that the sugarcane aphids preferred to settle on bloomless compared to wild-type plants. Overall, our results suggest that cuticular waxes on young sorghum leaves play a critical role in influencing host plant selection by sugarcane aphids.

Filter cake, a by-product in the sugar industry, has significant amount of wax containing policosanol which can be extracted using proper method and solvent. Policosanol is a mixture of high molecular weight aliphatic primary alcohols and was reported to reduce cardiovascular diseases. This study explored the use of hot ethanol reflux extraction to recover the crude wax from the filter cake followed by saponification for policosanol purification. Crude wax yield of 84.5 mg crude wax/g filter cake was obtained using reflux extraction at 60° C, 500 rpm and 12 mL absolute ethanol/g filter cake. The maximum crude wax yield obtained from Soxhlet extraction was 121.88 ± 0.46 mg crude wax/g dry filter cake, which corresponds to 69.33% recovery. Using a two-level factorial design, the effects of sodium hydroxide (NaOH) concentration (0.1 M and 1 M), reaction time (30 minutes and 60 minutes)and reaction temperature (30 °C and 60 °C) on the policosanol yield during saponification were determined. Results showed that NaOH concentration and temperature have significant effects on policosanol yield while reaction time alone is insignificant. Increasing both the reaction time and NaOH concentration can lead to an increase in policosanol yield. The highest yield of 1.234 ± 0.322 mg policosanol/g crude wax after saponification was obtained at 60 °C for 60 minutes with 1 M NaOH. This is twice the amount of policosanol recovered as compared to 0.583 ± 0.026 mg policosanol/g crude wax without saponification.

Aging-associated dyslipidemia and chronic low-grade inflammation contribute to atherosclerosis and cardiovascular risk. As a blend of long-chain aliphatic alcohols, policosanol from insect wax (PIW) has been documented to regulate lipid metabolism. However, the effects of PIW on atherosclerosis remain insufficiently characterized. In this study, ApoE mice fed a high-fat diet were concurrently administered PIW (75 and 150 mg/kg) for eight weeks. PIW was associated with weight gain reduction and improvement in lipid profile, particularly a decrease in triglycerides and total cholesterol. PIW also lowered circulating inflammatory biomarkers (IL-6, TNF-α, and C-reactive protein). Histopathological analyses revealed attenuated hepatic injury and reduced aortic lipid deposition and lesion features. In parallel, PIW reduced serum endothelin-1 and oxidized LDL levels and modulated aortic ET-1, MMP-9/TIMP-1 balance, and LOX-1/NF-κB-related protein signals. Notably, as PIW was administered concurrently with high-fat diet induction, these findings should be interpreted within a preventive intervention framework. Collectively, PIW help attenuate HFD-associated atherosclerotic features and hold promise as a functional food ingredient for cardiovascular health and healthy aging.

Insect wax is a famous biological resource for the role in economic production in China. Insect wax is a good source of policosanol, which may is a candidate supplement in foodstuff and pharmaceuticals that has important physiological activities. Therefore, this work aims to investigate a high-yield and rapid method for policosanol fabrication from insect wax. The conditions for policosanol fabrication were optimized as follows: an oil bath temperature of 112.7°C and reductant dosage of 0.97 g (used for the reduction of 10.00 g of insect wax). The yield of policosanol reached 83.20%, which was 4 times greater than that of existing methods, such as saponification. The total content of policosanol obtained under the optimal conditions reached 87%. In other words, a high yield of policosanol was obtained from insect wax (723.84 mg/g), that was 55 times higher than that generated from beeswax-brown via saponification. The concentrations of metal residues in policosanol were within the limits of the European Union regulations and EFSA stipulation. The LD50 values for oral doses of insect wax and policosanol were both > 5 g/kg. Policosanol was fabricated via solvent-free reduction from insect wax using LiAlH4 at a high yield. The fabrication conditions were optimized. Policosanol and insect wax showed high security, which made them potential candidates as supplements in foods, pharmaceuticals and cosmetics. The rapid and high-yield method has great potential for commercial manufacturing of policosanol.

Sugarcane and rice bran are the most important sources of commercial policosanol (PC) wax which exhibits a cholesterol lowering bioactivity. Both defatted rice bran (DRB) as agricultural waste and rice bran oil (RBO) retain a varying but significant amount of PC wax. Non-centrifugal cane sugar (NCS) has been consumed worldwide, and possesses various health benefits. It is mostly produced in hardened block form, which is not convenient for use compared with granular form. We aimed to increase PC contents of the granular sugar by adding wax extracted from DRB and RBO and to investigate the toxicity of the products. The results showed that the total PC contents including long chain aldehyde of products were increased to the maximum level of 147.97 mg/100 g. DRB is promising source of policosanol (6,044.7 mg/100 g). The main volatile components of developed sugar product was aldehyde and alcohol compounds. The 28 day toxicity evaluations of the developed sugar revealed no adverse effects.

ABSTRACT Lipases (EC 3.1.1.3) are triacylglycerol acylhydrolases that hydrolyze triglycerides to glycerol and fatty acids. The numerous applications of lipases in chemical, food, pharmaceutical, biotechnological, detergents and surfactants, and agricultural industries and in waste treatment plants proved their importance. In this study, 23 wax-degrading bacterial strains isolated from four mealybug species infesting cotton were screened for lipase production. On the basis of the 16S rRNA gene sequences, lipase-positive strains were classified into six genera, namely, Pseudoxanthomonas, Acinetobacter, Klebsiella, Providencia, Enterobacter, and Serratia. Acinetobacter lwoffii PSAD2 and A. beijerinckii PSAD7 isolates showed the maximum lipase production of 2.3 U/mL and 1.7 U/mL, respectively, in the quantification process. In addition, the growth curve analysis confirmed that lipase production is associated with the growth phase of the isolates. The efficacy of the lipase producing wax degrading bacterial isolates was tested against the nymphs and adults of Solenopsis Mealybug, Phenacoccus solenopsis Tinsley and the Striped Mealybug, Ferrisia virgata Cockerell infesting cotton. The results clearly indicated that the bacterial isolates A. lwoffii PSAD2 and A. beijerinckii PSAD7 expressed maximum mortality percentage against the mealybugs. The results of this study will be helpful in developing lipase-based microbial insecticides for the management of waxy cuticle-protected insect pests.

Background. Cuban sugarcane wax acids (SCWA) and policosanol (PCO) are mixtures of higher aliphatic acids and alcohols, respectively, purified from sugarcane wax with different chief components. Although it has been known that they have antioxidant and anti-inflammatory activities, physiological properties on molecular mechanism of SCWA have been less studied than PCO. Methods. In this study, we compared antiatherogenic activities of SCWA and PCO via encapsulation with reconstituted high-density lipoproteins (rHDL). Results. After reconstitution, SCWA-rHDL showed smaller particle size than PCO-rHDL with increase of content. PCO-rHDL or SCWA-rHDL showed distinct inhibition of glycation with similar extent in the presence of fructose. PCO-rHDL or SCWA-rHDL showed strong antioxidant activity against cupric ion-mediated oxidation of low-density lipoproteins (LDL), and inhibition of oxLDL uptake into macrophages. Although PCO-rHDL showed 1.2-fold stronger inhibition against cholesteryl ester transfer protein (CETP) activity than SCWA-rHDL, SCWA-rHDL enhanced 15% more brain cell (BV-2) growth and 23% more regeneration of tail fin in zebrafish. Conclusion. PCO and SCWA both enhance the beneficial functions of HDL to maximize its antioxidant, antiglycation, and antiatherosclerotic activities and the inhibition of CETP. These enhancements of HDL functionality by PCO and SCWA could exert antiaging and rejuvenation activity.

Context • Nonsteroidal, anti-inflammatory drugs effectively relieve osteoarthritis (OA) symptoms but also induce adverse effects (AEs) that limit their long-term use, which drives a search for safer treatments. D-002, a mixture of beeswax alcohols, and D-003, a mixture of sugarcane wax acids, have been effective in experimental and clinical studies for patients with OA. Objective • The study intended to investigate the effects on OA symptoms of a combined therapy using D-002 and D-003 (D-002/D-003), which were administered for 6 wk. Design • The study was a randomized, double-blind, placebo-controlled trial. Setting • The study was conducted at the Surgical Medical Research Center in Havana, Cuba. Participants • Participants were patients with mild-to-moderate OA. Intervention • Participants were randomly assigned to 1 of 4 groups-(1) a control group, which received a placebo; (2) the D-002 group (intervention group), which received 50 mg/d of D-002; (3) the D-003 group (intervention group), which received 10 mg/d of D-003; or (4) the D-002/D-003 group (intervention group), which received a combined therapy of 50 mg/d of D-002 plus 10 mg/d of D-003. The control group received tablets that were indistinguishable in appearance from the D-002 and D-003 tablets and had a similar composition, except that the active ingredients were replaced by lactose. The groups took the medications once per day for 6 wk. Outcome Measures • Symptoms were assessed using the Western Ontario and McMaster Individual Osteoarthritis Index (WOMAC) and a visual analogue scale (VAS). The primary outcome was the reduction in the total WOMAC score. The subscale scores on the WOMAC for pain, stiffness, and physical function, the VAS scores, and the use of rescue medications were secondary outcomes. Results • Of the 120 enrolled participants, 116 completed the study. The treatments with D-002, D-003, and D-002/D-003 reduced the mean total WOMAC scores significantly from baseline to postintervention, by 75.1%, 72.8%, and 91.2%, respectively. Compared with the placebo, the treatments decreased the mean WOMAC scores for pain, joint stiffness, and physical function significantly. The VAS scores significantly decreased, showing a 71.4%, a 66.9%, and an 84.7% reduction for the D-002, D-003, and D-002/D-003 groups, respectively. All the reductions were significant from the first week and were enhanced during the trial. The D-002/D-003 treatment was more effective in improving all of the scores than either monotherapy. With respect to rescue medications, 3/30, 2/30, and 2/30 used the medications in the D-002, D-003, and D-002/D-003 groups, respectively, vs 17/30 in the control group. The treatments were well tolerated. Conclusions • Administered for 6 wk, 50 mg/d of D-002 and 10 mg/d of D-003 ameliorated OA symptoms, but the combined therapy, D-002/D-003, was more effective than either monotherapy. All treatments were well tolerated.