Diet Therapy research studies for chronic conditions

Abstract
A potentially useful approach for drug discovery is to connect gene expression profiles of disease-affected tissues (“disease signatures”) to drug signatures, but it remains to be shown whether it can be used to identify clinically relevant treatment options. We analyzed coexpression networks and genetic data to identify a disease signature for type 2 diabetes in liver tissue. By interrogating a library of 3800 drug signatures, we identified sulforaphane as a compound that may reverse the disease signature. Sulforaphane suppressed glucose production from hepatic cells by nuclear translocation of nuclear factor erythroid 2–related factor 2 (NRF2) and decreased expression of key enzymes in gluconeogenesis. Moreover, sulforaphane reversed the disease signature in the livers from diabetic animals and attenuated exaggerated glucose production and glucose intolerance by a magnitude similar to that of metformin. Finally, sulforaphane, provided as concentrated broccoli sprout extract, reduced fasting blood glucose and glycated hemoglobin (HbA1c) in obese patients with dysregulated type 2 diabetes.

The effect of SFN-containing broccoli sprout extracts was studied in T2D patients
Prompted by these findings in vitro and in vivo, we set out to investigate the effects of SFN on glucose control in T2D patients. SFN has been provided at high concentration as broccoli sprout extracts (BSEs) in several clinical studies for cancer, chronic obstructive pulmonary disease, autism, and inflammatory diseases (www.clinicaltrials.gov) [we used high-performance liquid chromatography (HPLC)–purified SFN at 99.5% in our cell and animal studies, but this has not been used for human studies so far].

Here, we used a dried powder of an aqueous extract of broccoli sprouts, which contains high concentrations of glucoraphanin, the inert glucosinolate precursor of SFN.
Glucoraphanin is converted to SFN by the release of intrinsic sprout myrosinase during chewing and also by human enteric bacteria (37–40). After intake, the plasma concentration of SFN rises within 1 hour with a mean half-life of 1.77 ± 0.13 hours, but SFN exerts a sustained effect on gene expression (41). Renal tubular secretion is suggested to play a major role in the elimination (37). Safety studies using BSE corresponding to 50 to 400 ?mol SFN daily have shown that BSE is well tolerated without clinically significant adverse effects (42–45).
BSEs improves fasting glucose and HbA1c in obese patients with dysregulated T2D
We observed a clear association between HbA1c levels at start of treatment and ?HbA1c in response to BSE treatment (?HbA1c, 0.2 mmol/mol reduction per 1 mmol/mol higher HbA1c at start; P = 0.004, Fig. 4A), whereas there was no association in the placebo group (P = 0.5). There was also an association between BMI and ?HbA1c in BSE-treated patients (?HbA1c, 0.4 mmol/mol reduction per 1 kg/m2 higher BMI; P = 0.015 for the BSE group; not significant for the placebo group).

BSE is most effective in obese patients with dysregulated T2D
Glucose production is exaggerated in dysregulated T2D, which is reflected in the higher fasting blood glucose among the patients with dysregulated T2D in our study (8.6 ± 0.2 in patients with dysregulated T2D versus 7.5 ± 0.2 mM in patients with well-regulated T2D; P = 0.0001). Consequently, BSE reduced fasting glucose in patients with dysregulated T2D but not in patients with well-regulated T2D (P = 0.023). We also observed an association between BMI and BSE-induced change in HbA1c (P = 0.017), and HbA1c was significantly reduced after BSE treatment in obese patients with dysregulated T2D (P = 0.034; Fig. 4B). BSE was more effective in lowering fasting blood glucose in patients with elevated plasma triglyceride concentrations (P = 0.046 for the association between plasma triglycerides at study start and ?glucose; an inverted association was observed in placebo-treated patients; P = 0.008; Fig. 4D). It is also notable that BSE was more effective in lowering fasting blood glucose in patients with high HOMA-IR (P = 0.058 for the association between HOMA-IR and ?glucose; Fig. 4E), and the BSE-induced reduction of HbA1c correlated with high fatty liver index (P = 0.045; Fig. 4F).

No severe adverse effects of BSE were reported
Most patients tolerated the BSE well. Eight patients receiving BSE and seven patients receiving placebo reported gastrointestinal side effects such as loose stools and flatulence, typically present during the first few days of the treatment period, after which these symptoms disappeared. Ten BSE-treated and five placebo-treated patients reported mild respiratory infections, and there were also a few other reported adverse events, including orthopedic ailments, most likely unrelated to the study compound (table S5). Of the 103 patients, 6 (5 with BSE and 1 with placebo) discontinued the study because of nausea (2 patients), headache (1 patient), glucose above 15 mM (one of the exclusion criteria; 1 patient), hospital visit for suspected ileus (later successfully treated; 1 patient), and depression (1 patient on placebo) (table S6).
DISCUSSION
Together, our data show that SFN reduces glucose production, partly via NRF2 translocation and decreased expression of key gluconeogenetic enzymes, and that highly concentrated SFN administered as BSE improves fasting glucose and HbA1c in obese patients with dysregulated T2D. BSE was well tolerated, and SFN reduced glucose production by mechanisms different from that of metformin. SFN also protects against diabetic complications such as neuropathy, renal failure, and atherosclerosis in animal models because of its antioxidative effects (49–52).
Our data suggest that BSE has a direct effect on gluconeogenesis rather than hepatic insulin sensitivity, but the degree of IR may still influence the efficacy of BSE via altered constitutive NRF2 activity. It has been shown that insulin signaling activates NRF2 via phosphatidylinositide 3-kinase (53). Moreover, studies in cardiomyocytes have shown that NRF2 is activated at the early stages of T2D to protect against increased reactive oxygen species but is reduced at later stages of the disease (54). This is further supported by observations of reduced NRF2 expression in animals with IR (55, 56) and hepatic steatosis (27, 28).
It is not surprising that BSE was most effective in obese patients with dysregulated T2D. First, our animal experiments showed an effect of SFN on glucose control in metabolically dysregulated animals on a HFD but not in metabolically well-regulated animals on a low-fat diet. Second, gluconeogenetic rate was correlated with body weight in mice with diet-induced diabetes, and SFN reduced gluconeogenetic rate specifically in the heaviest mice. Third, hepatic glucose production is often exaggerated in patients with high HbA1c, whereas patients with low HbA1c primarily have an impairment of peripheral glucose uptake (47). Fourth, it has been shown that hepatic glucose production is increased particularly in obese T2D patients, potentially via elevated free fatty acids (34–36).
There is abnormal regulation of hepatic glucose production early in the development of T2D, but it is typically compensated for by increased insulin secretion (57). SFN has been shown to protect from pancreatic ? cell damage in animals (58). We observed no changes in insulin secretion, measured as HOMA-B and insulinogenic index, and BSE did not affect fasting glucose or HbA1c in well-regulated T2D patients. However, we observed that SFN prevented the development of hyperglycemia in diet-challenged rats, and it would be of interest to longitudinally study the long-term effects of BSE on glucose production and insulin secretion capacity in prediabetic individuals.
Glitazones and metformin were not ranked particularly high in the drug comparisons, suggesting that they do not affect the hepatic gene coexpression network that was associated with hyperglycemia in this case but exert their effects via other pathways. It is not entirely surprising because these drugs have different mechanisms of action from that of SFN.
It has been demonstrated that 1% [DCCT (Diabetes Control and Complications Trial) units] decrease of HbA1c corresponds to 37% reduced risk of microvascular complications (59). BSE treatment reduced HbA1c from 57.1 mmol/mol (or 7.38%) to 53.4 mmol/mol (or 7.04%) in obese patients with dysregulated T2D. The patients thereby reached the 7% treatment goal recommended by the American Diabetes Association (60), which is likely to represent a clinically meaningful effect.
Although the effect of BSE on glucose production was abolished in vitro when the conversion of glucoraphanin to SFN was prevented, we cannot fully determine that SFN explains the effect of BSE given to patients. High doses of BSE cannot yet be recommended to patients as a drug treatment but would require further studies, including data on which groups of patients would potentially benefit most from it. Finally, the findings provide support for using disease signatures based on coexpression networks to interrogate drug signatures, thereby using the large public repositories of gene expression data, as one of many strategies for repurposing compounds of immediate clinical relevance.

In recent decades, romaine lettuce has been one of the fastest growing vegetables with respect to its consumption and production. An understanding is needed of the effect of major phenolic phytochemicals from romaine lettuce on biological protection for neuron-like PC-12 cells. Phenolics in fresh romaine lettuce were extracted, and then its total phenolics and total antioxidant capacity were measured spectrophotometrically. Neuroprotective effects of phenolic extract of romaine lettuce and its pure caffeic acid derivatives (caffeic, chicoric, chlorogenic, and isochlorogenic acids) in PC-12 cells were evaluated using two different in vitro
METHODS:
lactate dehydrogenase release and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assays. Total phenolics and total antioxidant capacity of 100 g of fresh romaine lettuce averaged 22.7 mg of gallic acid equivalents and 31.0 mg of vitamin C equivalents, respectively. The phenolic extract of romaine lettuce protected PC-12 cells against oxidative stress caused by H(2)O(2) in a dose-dependent manner. Isochlorogenic acid, one of the phenolics in romaine lettuce, showed stronger neuroprotection than the other three caffeic acid derivatives also found in the lettuce. Although romaine lettuce had lower levels of phenolics and antioxidant capacity compared to other common vegetables, its contribution to total antioxidant capacity and antineurodegenerative effect in human diets would be higher because of higher amounts of its daily per capita consumption compared to other common vegetables.

Background:
The use of wheat grass (Triticum aestivum) juice for treatment of various gastrointestinal and other conditions had been suggested by its proponents for more than 30 years, but was never clinically assessed in a controlled trial. A preliminary unpublished pilot study suggested efficacy of wheat grass juice in the treatment of ulcerative colitis (UC).
METHODS:
A randomized, double-blind, placebo-controlled study. One gastroenterology unit in a tertiary hospital and three study coordinating centers in three major cities in Israel. Twenty-three patients diagnosed clinically and sigmoidoscopically with active distal UC were randomly allocated to receive either 100 cc of wheat grass juice, or a matching placebo, daily for 1 month. Efficacy of treatment was assessed by a 4-fold activity index that included rectal bleeding and number of bowel movements as determined from patient diary records, a sigmoidoscopic evaluation, and global assessment by a physician.
Results:
Twenty-one patients completed the study, and full information was available on 19 of them. Treatment with wheat grass juice was associated with significant reductions in the overall activity index (P=0.031) and in the severity of rectal bleeding (P = 0.025). No serious side effects were found. Fresh extract of wheat grass demonstrated a prominent tracing in cyclic voltammetry methodology, presumably corresponding to four groups of compounds that exhibit anti-oxidative properties.
Conclusion:
Wheat grass juice appeared effective and safe as a single or adjuvant treatment of active distal UC.

Background. Purple sweet potato leaves (PSPL) are widely grown and are considered a healthy vegetable in Taiwan. PSPL contain a high content of flavonoids, and the boiling water-extracted PSPL (PSPLE) is believed to prevent metabolic syndrome. However, its efficacy has not yet been verified. Therefore, we investigated the effect of PSPLE on adipocytes. Methods. The differentiated 3T3-L1 cells used in this study were derived from preadipocytes that were differentiated into adipocytes using an adipogenic agent (insulin, dexamethasone, and 3-isobutyl-1-methylxanthine); approximately 90% of the cells were differentiated using this method. Results. Treating the differentiated 3T3-L1 cells with PSPLE caused a dose-dependent decrease in the number of adipocytes rather than preadipocytes. In addition, treatment with PSPLE resulted in apoptosis of the differentiated 3T3-L1 cells as determined by DAPI analysis and flow cytometry. PSPLE also increased the expression of cleaved caspase-3 and poly ADP-ribose polymerase (PARP). Furthermore, PSPLE induced downregulation of interleukin-6 (IL-6) and tumor necrosis factor-? (TNF-?) gene expression in the differentiated 3T3-L1 cells. Conclusions. These results suggest that PSPLE not only induced apoptosis but also downregulated inflammation-associated genes in the differentiated 3T3-L1 cells.

Raspberries (Rubus idaeus L.) have been extensively studies worldwide because of their beneficial effects on health. Recently reports indicate that crude extracts of Rubus idaeus (RIE) have antioxidant and anticancer ability. The aim of this study was to evaluate the mechanism of its antimetastatic ability in oral cancer cells. In this study, SCC-9 and SAS oral cancer cells were subjected to a treatment with RIE and then analyzed the effect of RIE on migration and invasion. The addition of RIE inhibited the migration and invasion ability of oral cancer cells. Real time PCR, western blot and zymography analysis demonstrated that mRNA, protein expression and enzyme activity of matrix metalloproteinases-2 (MMP-2) were down-regulated by RIE. Moreover, the phosphorylation of Focal adhesion kinase (FAK), src, and extracellular signal-regulated kinase (ERK) were inhibited after RIE treatment. In conclusion, these results demonstrated that RIE exerted an inhibitory effect of migration and invasion in oral cancer cells and alter metastasis by suppression of MMP-2 expression through FAK/Scr/ERK signaling pathway.© 2016 Wiley Periodicals, Inc. Environ Toxicol, 2016.

This study compared the differences of two types of buckwheat sprouts, namely, common buckwheat ( Fagopyrum esculentum Moench) and tartary buckwheat ( Fagopyrum tataricum (L.) Gaertn.), in general composition, functional components, and antioxidant capacity. The ethanol extracts of tartary buckwheat sprouts (TBS) had higher reducing power, free radical scavenging activity, and superoxide anion scavenging activity than those of common buckwheat sprouts (CBS). As for chelating effects on ferrous ions, CBS had higher values than TBS. Rutin was the major flavonoid found in these two types of buckwheat sprouts, and TBS was 5 fold higher in rutin than CBS. The antioxidant effects of buckwheat sprouts on human hepatoma HepG2 cells revealed that both of TBS and CBS could decrease the production of intracellular peroxide and remove the intracellular superoxide anions in HepG2 cells, but TBS reduced the cellular oxidative stress more effectively than CBS, possibly because of its higher rutin (and quercetin) content.