1: Semin Cancer Biol. 2007 May 5; [Epub ahead of print] Links
Dietary histone deacetylase inhibitors: From cells to mice to man.Dashwood RH, Ho E.
Linus Pauling Institute, Department of Environmental & Molecular Toxicology, Oregon State University, Corvallis, OR 97331, USA.

Sulforaphane (SFN) is an isothiocyanate found in cruciferous vegetables, such as broccoli and broccoli sprouts. This anticarcinogen was first identified as a potent inducer of Phase 2 detoxification enzymes, but evidence is mounting that SFN also acts through epigenetic mechanisms. SFN has been shown to inhibit histone deacetylase (HDAC) activity in human colon and prostate cancer lines, with an increase in global and local histone acetylation status, such as on the promoter regions of P21 and bax genes. SFN also inhibited the growth of prostate cancer xenografts and spontaneous intestinal polyps in mouse models, with evidence for altered histone acetylation and HDAC activities in vivo. In human subjects, a single ingestion of 68g broccoli sprouts inhibited HDAC activity in circulating peripheral blood mononuclear cells 3-6h after consumption, with concomitant induction of histone H3 and H4 acetylation. These findings provide evidence that one mechanism of cancer chemoprevention by SFN is via epigenetic changes associated with inhibition of HDAC activity. Other dietary agents such as butyrate, biotin, lipoic acid, garlic organosulfur compounds, and metabolites of vitamin E have structural features compatible with HDAC inhibition. The ability of dietary compounds to de-repress epigenetically silenced genes in cancer cells, and to activate these genes in normal cells, has important implications for cancer prevention and therapy. In a broader context, there is growing interest in dietary HDAC inhibitors and their impact on epigenetic mechanisms affecting other chronic conditions, such as cardiovascular disease, neurodegeneration and aging.

PMID: 17555985 [PubMed - as supplied by publisher]

Deacetylase inhibitors increase muscle cell size by promoting myoblast recruitment and fusion through induction of follistatin.Iezzi S, Di Padova M, Serra C, Caretti G, Simone C, Maklan E, Minetti G, Zhao P, Hoffman EP, Puri PL, Sartorelli V.
Muscle Gene Expression Group, Laboratory of Muscle Biology, NIAMS, National Institutes of Health, Bethesda, MD 20892, USA.

Fusion of undifferentiated myoblasts into multinucleated myotubes is a prerequisite for developmental myogenesis and postnatal muscle growth. We report that deacetylase inhibitors favor the recruitment and fusion of myoblasts into preformed myotubes. Muscle-restricted expression of follistatin is induced by deacetylase inhibitors and mediates myoblast recruitment and fusion into myotubes through a pathway distinct from those utilized by either IGF-1 or IL-4. Blockade of follistatin expression by RNAi-mediated knockdown, functional inactivation with either neutralizing antibodies or the antagonist protein myostatin, render myoblasts refractory to HDAC inhibitors. Muscles from animals treated with the HDAC inhibitor trichostatin A display increased production of follistatin and enhanced expression of markers of regeneration following muscle injury. These data identify follistatin as a central mediator of the fusigenic effects exerted by deacetylase inhibitors on skeletal muscles and establish a rationale for their use to manipulate skeletal myogenesis and promote muscle regeneration.

PMID: 15130492 [PubMed - indexed for MEDLINE]

1: Nat Med. 2006 Oct;12(10):1147-50. Epub 2006 Sep 17. Links
Functional and morphological recovery of dystrophic muscles in mice treated with deacetylase inhibitors.Minetti GC, Colussi C, Adami R, Serra C, Mozzetta C, Parente V, Fortuni S, Straino S, Sampaolesi M, Di Padova M, Illi B, Gallinari P, Steinkühler C, Capogrossi MC, Sartorelli V, Bottinelli R, Gaetano C, Puri PL.
Dulbecco Telethon Institute at Fondazione A. Cesalpino, Institute of Cell Biology and Tissue Engineering, San Raffaele Biomedical Science Park of Rome, Via Castel Romano 100, 00128, Rome, Italy.

Pharmacological interventions that increase myofiber size counter the functional decline of dystrophic muscles. We show that deacetylase inhibitors increase the size of myofibers in dystrophin-deficient (MDX) and alpha-sarcoglycan (alpha-SG)-deficient mice by inducing the expression of the myostatin antagonist follistatin in satellite cells. Deacetylase inhibitor treatment conferred on dystrophic muscles resistance to contraction-coupled degeneration and alleviated both morphological and functional consequences of the primary genetic defect. These results provide a rationale for using deacetylase inhibitors in the pharmacological therapy of muscular dystrophies.

PMID: 16980968 [PubMed - indexed for MEDLINE]

If you want to know why this is interesting search for myostatin or folistation + histone deacetylase.

Very interesting indeed.
Curious thing though. HDAC inhibitors may stimulate revival of dormant herpes-1 virus, presumably leading to cold sores. Yet broccoli contains I3C in sufficient quantities to quash herpes-1.
It therefore follows that a meal with broccoli sprouts and mature broccoli will promote and prevent cold sores at the same time.
http://www.webmd.com/genital-herpes/news/2...rt-herpes-virus
http://www.pubmedcentral.nih.gov/articlere...i?artid=1361429

I realize this isn't related to making you hyoooge, and the data is confusing at best, but these factoids reminded me of an old Groucho Marx joke about pasta and sodium bicarbonate, so I couldn't resist mentioning it.

My god I'm a nerd.

When one speaks of HDAC, it refers to a class of enzymes. HDAC inhibitors, I assume, also refer to a number of agents.

My question is then, (1) does HDAC inhibitors that promote muscle growth act on SPECIFIC portions of our DNA? (2) And if so, does HDAC inhibitors from broccoli act on the same set of HDAC inhibitors?

1. Possible but from what I can tell the answer leans more towards no than yes. Several different HDAC inhibitors with verying profiles produce similar results as far as follistatin is conncerned. This of course is quite interesting in and of itself. It leads one to hypothesize that chromatin remodeling is the governer for satallite cell recuritment and myoblast fusion.

http://www.genesdev.org/cgi/content/full/18/21/2627

The Polycomb Ezh2 methyltransferase regulates muscle gene expression and skeletal muscle differentiation
Giuseppina Caretti1, Monica Di Padova1, Bruce Micales2, Gary E. Lyons2 and Vittorio Sartorelli1,3
1 Muscle Gene Expression Group, Laboratory of Muscle Biology, NIAMS, National Institutes of Health, Bethesda, Maryland 20892, USA; 2 Department of Anatomy, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA



Abstract

The Ezh2 protein endows the Polycomb PRC2 and PRC3 complexes with histone lysine methyltransferase (HKMT) activity that is associated with transcriptional repression. We report that Ezh2 expression was developmentally regulated in the myotome compartment of mouse somites and that its down-regulation coincided with activation of muscle gene expression and differentiation of satellite-cell-derived myoblasts. Increased Ezh2 expression inhibited muscle differentiation, and this property was conferred by its SET domain, required for the HKMT activity. In undifferentiated myoblasts, endogenous Ezh2 was associated with the transcriptional regulator YY1. Both Ezh2 and YY1 were detected, with the deacetylase HDAC1, at genomic regions of silent muscle-specific genes. Their presence correlated with methylation of K27 of histone H3. YY1 was required for Ezh2 binding because RNA interference of YY1 abrogated chromatin recruitment of Ezh2 and prevented H3-K27 methylation. Upon gene activation, Ezh2, HDAC1, and YY1 dissociated from muscle loci, H3-K27 became hypomethylated and MyoD and SRF were recruited to the chromatin. These findings suggest the existence of a two-step activation mechanism whereby removal of H3-K27 methylation, conferred by an active Ezh2-containing protein complex, followed by recruitment of positive transcriptional regulators at discrete genomic loci are required to promote muscle gene expression and cell differentiation.


[Keywords: Polycomb group; myogenesis; histone methylation; transcription]

Received July 20, 2004; revised version accepted September 7, 2004.



--------------------------------------------------------------------------------
By preventing inappropriate gene activation, transcriptional repression imposes unique patterns of gene expression and is essential for the specification and maintenance of cell identity (Francis and Kingston 2001). The Polycomb group (PcG) proteins repress transcription of the Drosophila Hox genes and participate in establishing the body anteroposterior axis (Simon et al. 1992). Whereas some PcG genes exert their activities at later stages of development, the PcG Enhancer of zeste, E(z), functions early in development by regulating expression of the gap genes (Pelegri and Lehmann 1994). The PcG coding sequences—and presumably their function—have been conserved throughout evolution. Both in the plant Arabidopsis thaliana and in the nematode Caenorhabditis elegans, the counterparts of Drosophila E(z) regulate homeotic gene expression (Goodrich et al. 1997; Ross and Zarkower 2003; Zhang et al. 2003). E(z) proteins are also involved in initiating X-chromosome inactivation (Plath et al. 2003) and in maintaining the epigenetic patters of pluripotent stem cells (Erhardt et al. 2003). In mammals, two E(z)-related genes have been isolated, Ezh1 and Ezh2 (Laible et al. 1997). Ezh1 expression is prevalent in the adult, whereas Ezh2 is expressed during embryonic development (Laible et al. 1997). Consistent with its expression pattern, Ezh2 is required for early mouse development. Ezh2-null mouse embryos die during the transition from pre- to postimplantation development (O'Carroll et al. 2001).
Among the PcG family, the E(z) proteins are unique in that they are chromatin-modifying enzymes with histone lysine methyltransferase (HKMT) activity (Cao et al. 2002; Czermin et al. 2002; Kuzmichev et al. 2002; Muller et al. 2002). Their catalytic activity resides in the evolutionarily conserved SET domain (Sims et al. 2003). Binding of Drosophila E(z) to a DNA Polycomb response element of the Ultrabithorax (Ubx) gene correlates with H3-K27 methylation and Ubx repression (Cao et al. 2002). Ezh2-mediated methylation of H3-K27 creates a docking site for the subsequent recruitment on the chromatin of the PRC1 (Polycomb repressive complex 1) complex containing additional PcG proteins (Czermin et al. 2002). The interaction of Ezh2 with the histone deacetylase HDAC1 suggests that both histone deacetylation and methylation converge to ensure transcriptional repression (van der Vlag and Otte 1999). The Ezh2 requirement for early mouse development has hampered the study of its role in regulating developmental and postnatal processes. However, a role for Ezh2 in cell cycle progression and cell differentiation has emerged from the analysis of several forms of aggressive tumors. Overexpression of Ezh2 has been reported in hormone-refractory, metastatic prostate cancers (Varambally et al. 2002) and in poorly differentiated and particularly aggressive breast carcinomas (Kleer et al. 2003). Resting cells derived from human lymphomas do not express Ezh2, but Ezh2 is strongly expressed in proliferating lymphoma cells (Visser et al. 2001). Pertinent to its putative role in cell differentiation are the findings that conditional inactivation of Ezh2 results in selectively impaired formation of pre-B and immature B cells but an unaltered development of pro-B cells (Su et al. 2003). Collectively, these and other (Bracken et al. 2003) findings suggest that Ezh2 may regulate cell growth and certain differentiation processes.

Because Ezh2 expression is developmentally regulated in skeletal muscle (Laible et al. 1997), we have tested the hypothesis that Ezh2 may be involved in controlling muscle gene expression and differentiation. Our results indicate that mouse skeletal muscle cells transduced with an Ezh2 retrovirus failed to undergo terminal differentiation and that this differentiation block was mediated by the SET domain, a region responsible for the HKMT activity. Ezh2 interacts with the DNA-binding protein YY1, and both proteins are found—along with the deacetylase HDAC1—on the regulatory regions of transcriptionally inactive muscle specific genes. Their presence correlated with H3-K27 methylation. Upon transcriptional activation, chromatin interaction of Ezh2, HDAC1, and YY1 was lost and replaced by the positive regulators of muscle transcription, SRF and MyoD. This molecular switch was accompanied by H3-K27 hypomethylation and histone hyperacetylation. Thus, our results indicate that the removal of an actively suppressing HKMT protein complex containing Polycomb Ezh2 and the subsequent engagement of positive transcriptional regulators characterize activation of muscle gene expression.

It leads one to hypothesize that chromatin remodeling is the governer for satallite cell recuritment and myoblast fusion.

It leads one to hypothesize that chromatin remodeling is the governer for satallite cell recuritment and myoblast fusion.

I'm not sure what it is that makes you now suggest that chromatin remodelling is important for differentiation. Thats the whole point of what makes eukaryotes so special regarding gene expression and the specialisation of tissues - without 'chromatin remodelling' you couldn't have it?

I can fuck up some broccoli. I typically eat half a bag to a bag of the frozen florets in a sitting. This is good news.

ohhh goddy someone who has done their homework.

1. Possible but from what I can tell the answer leans more towards no than yes.

When one speaks of HDAC, it refers to a class of enzymes. HDAC inhibitors, I assume, also refer to a number of agents.

My question is then, (1) does HDAC inhibitors that promote muscle growth act on SPECIFIC portions of our DNA?

Broccoli sprouts: An exceptionally rich source of inducers of enzymes that protect against chemical carcinogens

(chemoprotection / glucosinolates / isothiocyanates / sulforaphane / glucoraphanin)

Jed W. Fahey, Yuesheng Zhang, and Paul Talalay

Brassica Chemoprotection Laboratory and Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205

Contributed by Paul Talalay, July 3, 1997

Induction of phase 2 detoxication enzymes [e.g., glutathione transferases, epoxide hydrolase, NAD(P)H: quinone reductase, and glucuronosyltransferases] is a powerful strategy for achieving protection against carcinogenesis, mutagenesis, and other forms of toxicity of electrophiles and reactive forms of oxygen. Since consumption of large quantities of fruit and vegetables is associated with a striking reduction in the risk of developing a variety of malignancies, it is of interest that a number of edible plants contain substantial quantities of compounds that regulate mammalian enzymes of xenobiotic metabolism. Thus, edible plants belonging to the family Cruciferae and genus Brassica (e.g., broccoli and cauliflower) contain substantial quantities of isothiocyanates (mostly in the form of their glucosinolate precursors) some of which (e.g., sulforaphane or 4-methylsulfinylbutyl isothiocyanate) are very potent inducers of phase 2 enzymes. Unexpectedly, 3-day-old sprouts of cultivars of certain crucifers including broccoli and cauliflower contain 10-100 times higher levels of glucoraphanin (the glucosinolate of sulforaphane) than do the corresponding mature plants. Glucosinolates and isothiocyanates can be efficiently extracted from plants, without hydrolysis of glucosinolates by myrosinase, by homogenization in a mixture of equal volumes of dimethyl sulfoxide, dimethylformamide, and acetonitrile at -50°C. Extracts of 3-day-old broccoli sprouts (containing either glucoraphanin or sulforaphane as the principal enzyme inducer) were highly effective in reducing the incidence, multiplicity, and rate of development of mammary tumors in dimethylbenz(a)anthracene-treated rats. Notably, sprouts of many broccoli cultivars contain negligible quantities of indole glucosinolates, which predominate in the mature vegetable and may give rise to degradation products (e.g., indole-3-carbinol) that can enhance tumorigenesis. Hence, small quantities of crucifer sprouts may protect against the risk of cancer as effectively as much larger quantities of mature vegetables of the same variety.

Heating decreases epithiospecifier protein activity and increases sulforaphane formation in broccoli.

Matusheski NV, Juvik JA, Jeffery EH.

Department of Food Science and Human Nutrition, University of Illinois, 499 Bevier Hall, 905 South Goodwin Avenue, Urbana, IL 61801, USA.

Sulforaphane, an isothiocyanate from broccoli, is one of the most potent food-derived anticarcinogens. This compound is not present in the intact vegetable, rather it is formed from its glucosinolate precursor, glucoraphanin, by the action of myrosinase, a thioglucosidase enzyme, when broccoli tissue is crushed or chewed. However, a number of studies have demonstrated that sulforaphane yield from glucoraphanin is low, and that a non-bioactive nitrile analog, sulforaphane nitrile, is the primary hydrolysis product when plant tissue is crushed at room temperature. Recent evidence suggests that in Arabidopsis, nitrile formation from glucosinolates is controlled by a heat-sensitive protein, epithiospecifier protein (ESP), a non-catalytic cofactor of myrosinase. Our objectives were to examine the effects of heating broccoli florets and sprouts on sulforaphane and sulforaphane nitrile formation, to determine if broccoli contains ESP activity, then to correlate heat-dependent changes in ESP activity, sulforaphane content and bioactivity, as measured by induction of the phase II detoxification enzyme quinone reductase (QR) in cell culture. Heating fresh broccoli florets or broccoli sprouts to 60 degrees C prior to homogenization simultaneously increased sulforaphane formation and decreased sulforaphane nitrile formation. A significant loss of ESP activity paralleled the decrease in sulforaphane nitrile formation. Heating to 70 degrees C and above decreased the formation of both products in broccoli florets, but not in broccoli sprouts. The induction of QR in cultured mouse hepatoma Hepa lclc7 cells paralleled increases in sulforaphane formation.

Retention of Phytochemicals in Fresh and Processed Broccoli

# LENORA A. HOWARD11The authors are with the Dept. Of Food Science and Human Nutrition, Univ. Of Illinois at Urbana-Champaign, 268 Bevier Hall, 905 S. Goodwin Ave., Urbana, IL 61801. Address inquiries to Dr. Barbara P. Klein.,
# ELIZABETH H. JEFFERY11The authors are with the Dept. Of Food Science and Human Nutrition, Univ. Of Illinois at Urbana-Champaign, 268 Bevier Hall, 905 S. Goodwin Ave., Urbana, IL 61801. Address inquiries to Dr. Barbara P. Klein.,
# MATTHEW A. WALLIG11The authors are with the Dept. Of Food Science and Human Nutrition, Univ. Of Illinois at Urbana-Champaign, 268 Bevier Hall, 905 S. Goodwin Ave., Urbana, IL 61801. Address inquiries to Dr. Barbara P. Klein. and,
# BARBARA P. KLEIN

Our objective was to determine whether steam blanching, storage and preparation affected concentrations of sulforaphane (SF), sulforaphane nitrile (SFN), cyanohydroxybutene (CHB), iberin (I) or iberin nitrile (IN) in fresh and frozen broccoli. Broccoli (var. "Arcadia") was grown in St. Charles, IL over three seasons. Samples were steam blanched (2 min at 93 ± 5°C) within 24h of harvest, frozen and stored at −20°C up to 90 days, and fresh broccoli was stored at 4°C up to 21 days. Samples were analyzed uncooked or microwave cooked. SF, SFN, I, IN and CHB were determined by GC in dichloromethane extracts from lyophilized samples. Rates of loss for CHB and SF were similar during storage of fresh broccoli. Blanching, storage, and microwave cooking decreased (p < 0.01) concentrations of each compound in fresh and frozen broccoli.

Determination of sulforaphane in broccoli and cabbage by high-performance liquid chromatography

H. Lianga, Q.P. Yuana, Corresponding Author Contact Information, E-mail The Corresponding Author, H.R. Donga and Y.M. Liub
aDepartment of Pharmaceutical Engineering, Beijing University of Chemical Technology, Bei San Huan Dong Lu 15, Beijing 100029, China
bVegetables and Flowers Institute, China Academy of Agriculture Science, Beijing, China
Received 3 September 2004; revised 31 October 2005; accepted 15 November 2005. Available online 2 May 2006.



Abstract

Sulforaphane is an isothiocyanate that is present naturally in widely consumed cruciferous vegetables. A simple and accurate method for sulforaphane analysis based on methylene chloride extraction and RP-HPLC using linear gradient of acetonitrile in water was described. The feasibility of this procedure was tested by analysing the sulforaphane level in edible tissues of broccoli and cabbage as well as in other parts of broccoli. The mean value of sulforaphane content in broccoli was nearly five-fold higher than that in cabbage. The highest content of sulforaphane was found in the florets, while the lowest content was found in the leaves.

I have access to a 30 mg Sulforaphane product.

Steve did you end up experimenting with this?

I think it MIGHT combine well with Pea Protein, given the right flavor system.

I'm going to pick up a kilo of dehydrated broccoli powder to play with, both for this angle and for the alkalinization effects (since my veggie intake is close to nil). I think it MIGHT combine well with Pea Protein, given the right flavor system.