Dietary factors and the regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase: implications for breast cancer and development

A role for mevalonate in cancer development has long been suggested by findings that 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity is elevated in malignant cells. Increased synthesis mevalonate and mevalonate-derived nonsterol isoprenoids supports increased cell proliferation through the activation of growth-regulatory proteins and oncoproteins, and by promoting DNA synthesis. We have recently shown that mevalonate promotes the growth of human breast cancer cells both in culture and as tumors grown in nude mice. Inhibition mevalonate synthesis, therefore, may be an effective strategy to impair the growth of malignant breast cells. Several dietary compounds with known anti-cancer effects are also reported to inhibit HMG-CoA reductase activity. Here, we review evidence suggesting that inhibition of mevalonate synthesis may mediate the protective effects of cholesterol, plant isoprenoids, genistein, and long-chain n-3 polyunsaturated fatty acids (PUFAs) on experimental breast cancer.     

Yeast sequencing reports. Molecular cloning and sequencing of the hcs gene,which encodes 3-hydroxy-3-methylglutaryl coenzyme a synthase of Schizosaccharomyces pombe

We have cloned and sequenced the hcs gene, which is thought to encode a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase consisting of 447 amino acids, from the fission yeast Schizosaccharomyces pombe. The predicted amino acid sequence of the hcs product of S. pombe has homology with the HMG-CoA synthase of rat (47·8%), chicken (49·2%), hamster (47·1%) and human cells (46·9%). One of the hcs genes was replaced with a marker gene in the diploid cell. No viable hcs-disrupted haploid was isolated after tetrad dissection, suggesting that the hcs gene is essential for growth. However the hcs-defective mutant could be grown on a medium containing 5 mg/ml mevalonate. These results strongly support that the hcs gene encodes HMG-CoA synthase and S. pombe contains a single copy of the hcs gene. The sequence of the hcs gene has been entered into the public data libraries under Accession Number U32187.     

Effects of long-term supplementation of policosanol on blood cholesterol/glucose levels and 3-hydroxy-3-methylglutaryl coenzyme a reductase activity in a rat model fed high cholesterol diets

Food Science and Biotechnology - Policosanol is a well-defined nutraceutical for the management of blood cholesterol levels. The present study examined (i) the effect of policosanol supplementation...     

Effects of thymoquinone rich fraction and thymoquinone on plasma lipoprotein levels and hepatic low density lipoprotein receptor and 3-hydroxy-3-methylglutaryl coenzyme A reductase genes expression

The hypocholesterolemic effect of thymoquinone rich fraction (TQRF) extracted from Nigella sativa seeds using supercritical fluid extraction (SFE) in comparison with commercial available thymoquinone (TQ) in male Sprague–Dawley rats was investigated. Rats were fed prepared diet supplemented with 1% (w/w) cholesterol and treated with TQRF at, 0.5, 1 and 1.5 g/kg and TQ at 20, 50 and 100 mg/kg for 8 weeks. Plasma total cholesterol levels (TC) and low density lipoprotein cholesterol (LDLC) were significantly decreased in the TQRF and TQ treated rats compared to untreated rats. mRNA level of low density lipoprotein receptor (LDLR) was significantly expressed and the mRNA level of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-COAR) was significantly suppressed in the TQRF and TQ treated rats at different doses compared to untreated rats. These new findings identify TQRF and TQ as natural cholesterol lowering agents, and our study provides a molecular basis for the mechanisms of action through regulation of cholesterol in two main mechanisms first, uptake of LDLC via up regulation of LDLR gene and second, inhibition the synthesis of cholesterol via suppressing the HMG-COAR gene.     

共表达谷氨酰-tRNA还原酶增强染料脱色过氧化物酶在大肠杆菌中的表达活性

染料脱色过氧化物酶(dye-decoloorizing peroxidase,DyP)属于以血红素为辅基的新型过氧化物酶类,常因缺乏辅因子而导致催化活性低。将来源于褐色嗜热裂孢菌(Thermobifida fusca)的染料脱色过氧化物酶基因(TfuDyP)与大肠杆菌谷氨酰-tRNA还原酶基因(hemA),构建重组质粒phemA-DyP,转化至E.coli BL21中进行共表达。分别以2,2-联氮-二(3-乙基苯并噻唑-6-磺酸)二铵盐(ABTS)和顽固性染料活性蓝19(RB19)、溴酚蓝、溴甲酚绿为底物检测TfuDyP的催化活力以及染料脱色效率。结果表明,诱导后的共表达菌株pAD胞内血红素含量为9. 8μmol/L,而单独过表达基因TfuDyP的菌株pD仅为3. 4μmol/L。TfuDyP纯酶的全波长扫描分析表明,在菌株pAD中DyP酶与血红素的结合度相比pD有较大幅度的提升。pAD菌株表达的DyP酶活力较pD菌株提高了110%,酶活力的提高使其在染料脱色应用方面也得到增强。在pAD菌株培养基中分别添加谷氨酸(Glu)、FeCl2使得胞内血红素含量、DyP酶活力和染料脱色效率比未添加时进一步提高。以上结果为TfuDyP的功能开发奠定了基础,同时也为其他血红素依赖性过氧化物酶的研发提供借鉴。     

Identification of phenolic compounds isolated from pigmented rices and their aldose reductase inhibitory activities

Two anthocyanins (cyanidin-3-O-β-glucoside and peonidin-3-O-β-glucoside) and other phenolic (ferulic acid) were, respectively isolated from black and pigmented brown rices (Oryza sativa L. japonica) and their complete structures were determined by spectroscopic analyses (H NMR, C NMR and MALDI MASS). The HPLC profile of anthocyanins extracted from black rice showed cyanidin-3-O-β-glucoside as the first peak (85%) and peonidin 3-O-β-d-glucoside as the second (15%), while that of pigmented brown rice showed ferulic acid as the first peak (85.7%) and tocols as the second (14.3%). Several tocols were isolated and identified from the unsaponifiable fractions of both rices having some difference on their structures and amounts. The aldose reductase inhibitory activity of isolated compounds was in the following decreasing order: cyanidin-3-glucoside > quercetin > ferulic acid > peonidin-3-glucoside > tocopherol.     

Screening of edible mushrooms and extraction by pressurized water (PWE) of 3-hydroxy-3-methyl-glutaryl CoA reductase inhibitors

The methanol/water and particularly the water extracts obtained from 26 mushroom species were able to inhibit the 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR) activity to different extent (10–76%). Cultivated mushrooms such as Pleurotus sp. and Lentinula edodes were among the strains which showed higher HMGCR inhibitory capacities. Their inhibitory properties were not largely influenced by cultivation parameters, mushroom developmental stage or flush number. The HMGCR inhibitory activity of L. edodes was concentrated in the cap excluding the gills while in Pleurotus ostreatus it was distributed through all the different tissues. A method to obtain aqueous fractions with high HMGCR inhibitory activity was optimized using an accelerated solvent extractor (ASE) by selecting 10.7 MPa and 25 °C as common extraction conditions and 5 cycles of 5 min each for P. ostreatus fruiting bodies and 15 cycles of 5 min for L. edodes suggesting that the potential HMGCR inhibitors are different in the two selected mushrooms.     

Bitterness of cassava: Identification of a new apiosyl glucoside and other compounds that affect its bitter taste

Compounds extracted by methanol from cassava parenchyma and cortex have been separated on a preparative HPLC column and identified by ¹H and ¹³C NMR spectroscopy. A new compound isopropyl-β-D -apiofuranosyl-(1 → 6)-β-D -glucopyranoside (IAG, structure I ) has been found as well as small amounts of phenylalanine and tryptophan. The composition of another HPLC fraction has not been elucidated. The amounts of the identified compounds and of linamarin, lotaustralin, citrate, malate and the various sugars present in cassava have been determined by HPLC methods. The threshold levels of bitterness of aqueous solutions of linamarin, lotaustralin and IAG, have been determined and together with published data on L -phenylalanine and L -tryptophan have allowed our evaluation of their contributions to the bitterness of cassava. Linamarin is the sole contributor in the parenchyma but (with two cultivars out of six studied) IAG contributes more to the bitterness of the cortex than does linamarin. The perception of bitterness of linamarin solutions is confounded in the presence of neutral citrate and malate which have a sour taste. These modify the taste of cassava tubers. There are many compounds that contribute to the taste of cassava tubers, hence it is not surprising that the bitterness of cassava is not always correlated positively with the cyanide potential.     

Investigations of anti-inflammatory,antinociceptive, antioxidant and aldose reductase inhibitory activities of phenolic compounds from Sideritis brevibracteata

The Sideritis species are widely used in the treatment of gastrointestinal disorders, rheumatism, cough, the common cold and as a diuretic, and they are commonly used as an herbal tea in folk medicine in Turkey. The teas of Sideritis have unique tastes, pleasant aromas and a transparent yellow colour, and are preferred as a hot beverage. In order to evaluate this information, the crude methanolic extract and the water soluble part of the methanolic extract after partition with chloroform and n-butanol were investigated for their anti-inflammatory and antinociceptive activities. Phytochemical studies performed on the n-butanol extract of the overground parts of Sideritis brevibracteata, which is rich in phenolic compounds with promising activities, resulted in the isolation of six acetylated allose containing 8-hydroxyflavone glycosides and a phenylethanoid glycoside. The structures of the compounds were established by spectroscopic evidence (UV, IR, NMR, MS). Furthermore, these phenolic compounds of S. brevibracteata were studied for anti-inflammatory, antinociceptive, antioxidant and aldose reductase inhibitory activities. The experimental data demonstrated that S. brevibracteata displayed remarkable anti-inflammatory, antinociceptive and antioxidant activities.     

Comparative effects of safflower oil and perilla oil on serum and hepatic lipid levels, fatty acid compositions of serum and hepatic phospholipids, and hepatic mRNA expressions of 3-hydroxy-3-methylglutaryl CoA reductase, LDL receptor, and cholesterol 7alpha-hydroxylase in young and adult rats

Male Wistar rats, 4 and 33 weeks of age, were fed the diets containing safflower oil (SO-diet, 77.3% linoleic acid) or perilla oil (PO-diet, 58.4% -linolenic acid) for 7 days. Serum total cholesterol was lower on the PO-diet in both ages. On the other hand, hepatic cholesterol and phospholipids were significantly higher in the PO group than in the SO group of the adult rats. The PO group showed significantly lower 20:4 n-6 but higher 18:2 n-6 in hepatic phosphatidylcholine compared with the SO group in both ages. Hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase mRNA was significantly lower on the PO-diet than on the SO-diet irrespective of age. The present results show that -linolenic acid has a higher hypocholesterolemic ability than linoleic acid in rats irrespective of age and these fatty acids behaved differently in affecting hepatic mRNA expressions of HMG-CoA reductase, LDL receptor, and cholesterol 7-hydroxylase.     

Site-directed mutagenesis improves the thermostability of a recombinant Picrophilus torridus trehalose synthase and efficiency for the production of trehalose from sweet potato starch

A new recombinant Picrophilus torridus TSase (PTTS) has the catalytic ability for the conversion of maltose to trehalose by intramolecular transglucosylation. For industrial applications, the high thermostability of the enzyme would be the most important property for reducing the microbial contamination and lower the production cost. Therefore, in this study, we substituted ten selected proline residues of PTTS which differ from two well-known thermostable TSases. Interestingly, we found that the N503 mutant type, namely N503P-PTTS, showed about 39% higher relative activity than that of the wild type at 65 °C for 120 min. The trehalose yield of mutant N503P-PTTS was 1.3-fold higher than that of the wild type with sweet potato starch as substrate at 50 °C for 4 h. This suggests that the proline site substitution technology used in this study is useful for altering enzyme properties and catalytic efficiency for possible industrial applications.     

Occurrence of 3-chloro-propane-1,2-diol (3-MCPD) and related compounds in foods: a review

A critical review of the occurrence of 3-chloro-propane-1,2-diol (3-MCPD) in foods not known to contain hydrolysed vegetable proteins is presented. The review covers the properties and chemistry of 3-MCPD and the current methods of analysis in foodstuffs. The results of UK surveys of 3-MCPD occurrence in both retail foods and commercial food ingredients are discussed with particular reference to cereal, meat and dairy products. The possible mechanisms for the formation and decay of 3-MCPD in foods are suggested. The review does not cover the detailed toxicology of 3-MCPD and its occurrence in hydrolysed vegetable proteins, which have been considered elsewhere, nor possible issues such as in-vivo formation.     

Antioxidant activities of compounds isolated from Dalbergia odorifera T. Chen and their inhibition effects on the decrease of glutathione level of rat lens induced by UV irradiation

Four compounds were isolated from the root of Dalbergia odorifera T. Chen and identified by chemical and spectroscopic methods as 2′-O-methyl-isoliquiritigenin (1), odoriflavene (2), 5′-methoxy-vestitol (3) and formononetin (4). Their antioxidant activities were investigated in lard by the oxidative stability instrument; their inhibitory effects on the decrease of glutathione level of rat lens induced by UV irradiation were studied using a rat lens UV-damage model, and their in vitro cytotoxicity was measured on human SH-SY5Y neuroblastoma cells. Results indicated that all four compounds had obvious antioxidant effects, while none of them showed any synergistic effects on butylated hydroxytoluene or α-tocopherol. When FeCl₃ (4.0 μM) was added, the antioxidant activities of the four compounds markedly decreased. The inhibitory effects of compounds 1, 2 and 4 on the decrease of glutathione level of rat lens induced by UV irradiation were comparable to that of α-tocopherol. All four compounds showed cytotoxic activity when evaluated against a SH-SY5Y cell line in vitro.     

Effect of ellagitannins,ellagic acid and volatile compounds from oak wood on the (+)‐catechin,procyanidin B1 and malvidin‐3‐glucoside content of model wines

Background and Aims: During ageing in oak barrels, wine undergoes changes because of the release of polyphenols and other molecules from wood. The aim of this study was to evaluate the influence of some oak wood‐derived volatile compounds, ellagic acid and oak wood extracts on the levels of (+)‐catechin, procyanidin B1 and malvidin‐3‐glucoside. Methods and Results: Phenolics and the oak wood derived volatile compounds studied were quantified by HPLC and by GC, respectively. Additionally, the new compounds formed in the solutions were characterised by their spectral properties. Ellagic acid and/or oak wood extracts slowed the decline in the levels of (+)‐catechin and procyanidin B1. In contrast, the decrease in malvidin‐3‐glucoside was more pronounced in the presence of ellagic acid and oak wood chip extracts. Furfural slowed (+)‐catechin degradation, while breakdown of malvidin‐3‐glucoside was slightly more pronounced in the presence of guaiacol, furfural, vanillin and eugenol. (+)‐Catechin, procyanidin B1 and malvidin‐3‐glucoside did not significantly affect the rate of the degradation of ellagitannins during the storage time studied. Finally, new HPLC peaks were detected in the solutions containing (+)‐catechin and ellagic acid, as well as with malvidin‐3‐glucoside with ellagic acid and oak wood extract. Conclusions: Malvidin 3‐glucoside and (+)‐catechin and procyanidin B1 presented distinct behaviours during time in the presence of volatile and non‐volatile compounds from oak wood. Significance of the Study: This work points out the importance of oak wood components in the degradation of anthocyanins and tannins, as well as the reactions that occur during the ageing of red wine.     

Structure and pesticidal activities of derivative of dinitrophenols. 3. Structure and acaricidal activity of certain carbonates of 2-alkyl-5-methyl-4,6-dinitrophenols and related compounds

The activities of alkyl 2-alkyl-5-methyl-4,6-dinitrophenyl carbonates and of related compounds against Tetranychus telarius (greenhouse red spider mite) were examined. The 5-methyl substituent slightly reduced the toxicity to mites, but not to mammals. The carbonates of 2-isopropyl- and 2-s-butyl-5-methyl 4,6-dinitrophenols were more acaricidal than the parent phenols. The increase in chain length of the alkoxyl group in the carbonates caused a decrease in acaricidal activity. In contrast to the branched carbonates of 2-t-butyl-4,6-dinitrophenol their 5-methyl-analogues were as active as the normal homologues. The cyclohexyl and phenyl carbonates of the 5-methyl-analogues had better activity than those without the 5-methyl substituent. The thiolo- and dithio-carbonates were found to be less active than their oxy-analogues.     

Isolation of methyl syringate as a specific aflatoxin production inhibitor from the essential oil of Betula alba and aflatoxin production inhibitory activities of its related compounds

Methyl syringate was isolated from the essential oil of Betula alba as an aflatoxin production inhibitor. It inhibited aflatoxin production of Aspergillus parasiticus and Aspergillus flavus with IC₅₀ values of 0.9 and 0.8 mM, respectively, without significantly inhibiting fungal growth. Methyl syringate reduced mRNA levels of genes (aflR, pksA, and omtA) encoding proteins required for aflatoxin biosynthesis. Methyl gallate, methyl 3,4,5-trimethoxybenzoate, and methyl 3-O-methylgallate inhibited both aflatoxin production and fungal growth of A. parasiticus and A. flavus. However, their acids and syringic acid did not inhibit aflatoxin production and growth of A. parasiticus significantly, although gallic acid inhibited aflatoxin production of A. flavus with selectivity. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of methyl syringate was much weaker than that of gallic acid. These results showed that methyl syringate has a unique inhibitory activity toward aflatoxin production with a different mode of action from that of gallic acid.