BACKGROUND:Brain tumors are highly aggressive tumors characterized by secretions of high levels of matrix metalloproteinase-2 and -9, leading to tumor growth, invasion and metastasis by digesting the basement membrane and extracellular matrix components. We previously demonstrated the effectiveness of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, and green tea extract in vitro: on activity of urokinase plasminogen activator, matrix metalloproteinases and TIMPs in various human glioblastoma (LN-18, T-98G and A-172) cell lines and on glioblastoma A-172 cell proliferation and Matrigel invasion.

AIM:Our main objective in this study was to investigate the effect of the NM in vivo on human glioblastoma U-87 MG cell line.

MATERIALS AND METHODS:Athymic male nude mice inoculated with 3·10(6) U-87 MG cells subcutaneously and were fed a regular diet or a regular diet supplemented with 0.5% NM. Four weeks later, the mice were sacrificed, the tumors were weighed and measured. The samples were studied histologically.

RESULTS:NM inhibited tumor weight and tumor burden by 53% (p = 0.015) and 48% (p = 0.010), respectively.

CONCLUSIONS:These results suggest the therapeutic potential of NM as an adjuvant in the treatment of glioblastoma.

Ovarian cancer is the deadliest gynecological malignancy in women, and fifth leading cause of death. Despite advances made in chemotherapy and surgery, the average time of clinical remission is approximately 2 years and the 5-year survival rate is 45%. Thus, there is an urgent need for the development of a novel therapeutic approach to ovarian cancer treatment. We investigated the effect of a specific nutrient mixture (EPQ) containing ascorbic acid, lysine, proline, green tea extract, and quercetin on human ovarian cancer cell A-2780 in vivo and in vitro. Athymic female nude mice (n = 12) were all inoculated intraperitoneally (IP) with 2× 10⁶ cells in 0.1 mL of phosphate buffered saline (PBS) and randomly divided into two groups. Upon injection, the Control group (n = 6) was fed a regular diet and the EPQ group (n = 6) a regular diet supplemented with 0.5% EPQ. Four weeks later, the mice were sacrificed and tumors that developedin the ovary were excised, weighed, and processed for histology. Lungs were inspected for metastasis. In vitro, A-2780 cells were cultured in Dulbecco modified Eagle medium supplemented with 10% FBS and antibiotics. At near confluence, cells were treated with EPQ in triplicate at concentrations between 0 and 1000 μg/mL. Cell proliferation was measured via MTT assay, MMP-9 secretion via gelatinase zymography, invasion through Matrigel and morphology via hematoxylin and eosin (H& E) staining. All Control mice developed large ovarian tumors, whereas 5 out of 6 mice in the EPQ group developed no tumors, and one, a small tumor. Control mice also showed lung metastasis in 6 out of 6 mice, while no lung metastasis was evident in EPQ mice. Zymography demonstrated only MMP-9 expression, which EPQ inhibited in a dose-dependent fashion, with virtual total block at 250μg/mL concentration. EPQ significantly inhibited invasion through Matrigel with total block at 250 μg/mL concentration. MTT showed dose-dependent inhibition of cell proliferation with EPQ, and H& E staining showed no morphological changes below 500μg/mL EPQ. These results suggest that EPQ has therapeutic potential in the treatment of ovarian cancer by significantly suppressing ovarian tumor incidence and growth and lung metastasis, and by inhibiting MMP-9 secretion and invasion of A-2780 ovarian cancer cells.

INTRODUCTION:Aerobic training and green tea extract can be used to reduce the risk of prostate cancer. The goal of this study was to evaluate the effects of eight-week aerobic exercise training and administration of green tea extract on the level of nuclear factor kappa B (NF-kB), cyclooxygenase-2 (COX-2) and p53 tumor suppressor protein (p53) in prostate of rats which were stimulated by N-Methyl-N-nitrosourea (NMU) to induce the prostate cancer.

METHODS:60 adult male Wistar rats were assigned into six groups including healthy control (HCt), cancer control (CCt), cancer training (CTr: 45 min/day at low-moderate intensity, 5 times/week, 8 weeks), cancer extract (CEx: 1.34 ml of green tea extract, 3 times/week, 8 weeks), cancer training+ cancer extract (CTr+CEx) and sham groups. Rats were sacrificed 48 hours after the last intervention session, and the prostate tissue was isolated to measure the levels of NF-kB, COX-2, and p53.

RESULTS:The NF- kB level in CCt group was increased significantly compared to the HCt (P=0.02). In the CTr group, NF-kB level was decreased significantly compared to the CCt and CEx groups (P=0.001 and 0.05, respectively). In addition, the levels of P53 protein were reduced in CTr, CEx and CTr+CEx groups compared to CCt group (P=0.001, 0.02 and 0.004, respectively). No significant changes were found in the level of COX-2 between groups.

CONCLUSIONS:These results suggest that a long-term exercise training combined with the intake of green tea extract may reduce levels of NF-kB and p53 in rats with prostate cancer. Given the importance of recognizing complementary therapies in this regard, future studies are warranted.

BACKGROUND:Low-grade chronic inflammation in overweight subjects is thought to play an important role in disease development.

OBJECTIVE:It was hypothesized that specific dietary components are able to reduce low-grade inflammation as well as metabolic and oxidative stress.

DESIGN:Dietary products [resveratrol, green tea extract, alpha-tocopherol, vitamin C, n-3 (omega-3) polyunsaturated fatty acids, and tomato extract] selected for their evidence-based antiinflammatory properties were combined and given as supplements to 36 healthy overweight men with mildly elevated plasma C-reactive protein concentrations in a double-blind, placebo-controlled, crossover study with treatment periods of 5 wk. Inflammatory and oxidative stress defense markers were quantified in plasma and urine. Furthermore, 120 plasma proteins, 274 plasma metabolites (lipids, free fatty acids, and polar compounds), and the transcriptomes of peripheral blood mononuclear cells and adipose tissue were quantified.

RESULTS:Plasma adiponectin concentrations increased by 7%, whereas C-reactive protein (principal inflammation marker) was unchanged. However, a multitude of subtle changes were detected by an integrated analysis of the "omics" data, which indicated modulated inflammation of adipose tissue, improved endothelial function, affected oxidative stress, and increased liver fatty acid oxidation.

CONCLUSION:An intervention with selected dietary products affected inflammatory processes, oxidative stress, and metabolism in humans, as shown by large-scale profiling of genes, proteins, and metabolites in plasma, urine, and adipose tissue. This trial was registered at clinical trials.gov as NCT00655798.

Certain drastic behavioral modifications by arterial wall smooth muscle cells (SMC) have been considered key steps in the formation of atherosclerotic lesions: massive migration of SMC from the media to the intima layer of the vessel, dedifferentiation of SMC to proliferating phenotype, and increased secretion of inflammatory cytokines as a response to inflammatory stimuli. We investigated the anti-atherogenic effects of naturally occurring compounds (ascorbic acid, green tea extract, lysine, proline, arginine, and N-acetyl cysteine) using the model of cultured aortic SMC. Cell growth was measured by DNA synthesis, cell invasiveness was measured through Matrigel, matrix metalloproteinase-2 (MMP-2) secretion was measured by zymography, and SMC secretion of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) was measured by immunochemistry. Fetal bovine serum-stimulated SMC growth was inhibited by the nutrient mixture (NM) with 85% inhibition at 100 microg/mL. A corresponding concentration of epigallocatechin gallate (EGCG; 15 microM), the most active tea phenolic, produced a significant effect but one lower than NM. NM inhibited aortic SMC Matrigel invasion in a dose-dependent manner and significantly decreased MMP-2 expression. Stimulation of SMC with tumor necrosis factor-alpha significantly increased production and secretion of such mediators of inflammation as IL-6 and MCP-1; addition of 100 microg/mL NM inhibited secretion of MCP-1 and IL-6 by 65% and 47%, respectively. These data suggest that the NM of ascorbic acid, tea phenolics, and selected amino acids has potential in blocking the development of atherosclerotic lesions by inhibiting atherogenic responses of vascular SMC to pathologic stimuli and warrants in vivo studies.

Non-Hodgkin lymphomas incidence has increased more than 70% in last 25 years. Aggressiveness, higher relapse rate, and treatment complications pose significant barriers. Decreased food intake and side effects of treatments make cancer patients vulnerable to deficiency of essential nutrients such as vitamin C, lysine, and proline leading to the formation of weak extra cellular matrix susceptible to easy breakdown by matrix metalloproteinase enzymes. Inhibition of these enzymes has shown promise in stopping metastasis. Aim: In this study, we investigated the effects of a specific nutrient mixture, containing ascorbic acid, lysine, proline, green tea extract among others, in most aggressive forms of non-Hodgkin's lymphoma - Burkitt's lymphoma, and T-cell lymphoma - using Raji and Jurkat cells respectively. Methods: Nutrient mixture (NM) doses of 0, 10, 50, 100, 500, 1000 microg/ml, were used to study effects on cell proliferation, expression of matrix metalloproteinase, Matrigel invasion and apoptosis. Results: The results demonstrated that the dose response toxicity of the nutrient mixture on Raji cells gradually increased with increasing concentration. The nutrient mixture was non-toxic to Jurkat cells, however exhibited anti-proliferative properties at higher concentrations. Zymography demonstrated, NM had a significant inhibitory effect on matrix metalloproteinase-9 expression with total inhibition at 1000 microg/ml for Raji cells and at 500 microg/ml for Jurkat cells. The NM at 100 microg/ml completely inhibited Matrigel invasion for Raji cells, and at 1000 microg/ml for Jurkat cells. After the NM challenge virtually all Raji and Jurkat cells exposed to 1000 microg/ml were in late apoptosis. Conclusion: Considering the lack of treatment options and continually increasing incidence, NM could be further explored for its therapeutic potential in Burkitt's lymphoma and T-cell lymphoma.

BACKGROUND: Current treatment of pancreatic cancer is generally associated with poor prognosis, even if diagnosed early, owing to its aggressive rate of metastasis and non-responsiveness to chemotherapy and radiotherapy. Matrix metalloproteinases (MMPs) have received much attention in recent years for their role in various malignancies, and have been implicated in tumor invasion, metastasis, and angiogenesis. AIM OF STUDY: Reported antitumor properties of ascorbic acid, lysine, proline, and green tea extract prompted us to investigate the effect of a combination of lysine, proline, arginine, ascorbic acid, and green tea extract on pancreatic cancer cell line MIA PaCa-2 for viability, MMP expression, invasion, and morphology. METHODS: Viability was evaluated based on cell proliferation by MTT assay and MMP expression in condition media by gelatinase zymography. Invasion through Matrigel was assayed and morphology was observed by hematoxylin and eosin (H+E)staining. Data was analyzed by independent sample "t" test. RESULTS: The nutrient mixture (NM) did not inhibit cell proliferation at 10 microg/mL and exhibited a dose-dependent antiproliferative effect with maximum inhibition of 38% over the control at 1000 microg/mL. Zymography demonstrated production of only MMP-9, which showed a dose-dependent decreased expression that was abolished at 100 microg/mL of NM. Invasion through Matrigel was inhibited at 10, 50, 100, and 500 microg/mL by 66%, 66%, 87% and 100%, respectively. H&E staining did not indicate changes even at the highest concentration of NM. CONCLUSION: Our results suggest that the formulation of green tea extract, lysine, proline, and ascorbic acid, tested as a promising adjunct to standard treatment of pancreatic cancer, by inhibiting MMP expression and invasion without toxic effects important parameters in cancer metastasis.

We have previously reported that decaffeinated green tea extract (GTE) in combination with voluntary exercise (Ex) reduces metabolic syndrome in high fat-fed C57BL/6J mice. Here, we examined for the first time the effect of treatment with 77 mg/g GTE, Ex, or both (GTE + Ex) on genes related to the conversion of white adipose tissue (WAT) to brown fat-like adipose tissue (BLAT) in this model. GTE+Ex induced genes related to lipolysis (hormone sensitive lipase [3.0-fold] and patatin-like phospholipase domain-containing protein 2 [2-fold]), mitochondrialβ-oxidation (NADH dehydrogenase 5 [2.3-fold], cytochrome B [2.0-fold], and cytochrome C oxidase III [1.9-fold increase]), and adipose tissue browning (peroxisome proliferator-activated receptor-γ coactivator-1α [1.8-fold], bone morphogenetic protein 4 [2.6-fold], and phosphatase and tensin homolog [2.6-fold]) in visceral WAT compared to HF-fed mice. These results suggest that GTE+Ex function in part by inducing the conversion of WAT to BLAT and provides novel mechanistic insight into this combination.

AIMS:Green tea extract (GTE) can exert anti-obesity and inflammatory effects. Our study determined whether the benefits of GTE are summative with exercise-induced changes in anthropometric indices, and the levels of inflammatory cytokines, adiponectin and irisin in inactive overweight women.

METHODS:Thirty overweight female participants were randomized to three groups: endurance training + placebo (ET+P); endurance training + GTE (ET +GTE); and Control (no exercise) + placebo (Control, N=10). The exercise intervention consisted of an eight-week endurance-training program of three sessions per week [aerobics, aerobic circuit training, and fast walking or jogging at a moderate intensity of 40-59% of the heart rate reserve]. The dose of GTE used was 500 mg/d in the form of a green tea capsule.

RESULTS:Body weight, body mass index (BMI), waist to hip ratio (WHR), and body fat percentage (BFP) were decreased in both ET+P and ET+GTE interventions (P<0.001 for both interventions). The reduction of anthropometric values in the ET+GTE group was significantly higher than ET+P interventions (P<0.001). Both exercise interventions also significantly (P<0.001) increased adiponectin [ET +GTE= 5.28 mg/ml (95% CI, 4.48 to 6.08) and ET+P= 3.34 mg/ml (95% CI, 2.76 to 3.92)] and decreased hs-CRP [ET +GTE= -0.95 mg/l (95% CI, -1.15 to -0.75) and ET+P= -0.35 mg/l (95% CI, -0.46 to -0.24)]. Changes in adiponectin and hs-CRP were greater (P<0.05) in ET+GTE compared to ET+P. There were no significant differences in irisin, IL-6, and TNF-α between the three groups (P>0.05).

CONCLUSIONS:GTE improves exercise-induced body composition by further decreasing exercise-induced changes in weight, BMI, WHR, and BFP. The combination of GTE and exercise also produced greater changes in anti-inflammatory (increases in adiponectin) and metabolic (decreases in hs-CRP) markers than exercise alone.

Cervical cancer is one of the most commonly diagnosed cancers and a significant cause of mortality in women worldwide. Although cervical cancer is fully treatable in the early stages, once it has metastasized, patient outcome is poor. The objective of the present study was to investigate the effect of dietary supplementation with a nutrient mixture (NM) containing lysine, ascorbic acid, proline, green tea extract and other micronutrients on the expression of extracellular matrix (ECM) proteins in HeLa cell xenografts in nude female mice. After housing for 1 week, female athymic nude mice between 5 and 6 weeks of age (n=12) were inoculated subcutaneously with 3×10(6) HeLa cells in phosphate-buffered saline and Matrigel and randomly divided into two groups. These were the control group, in which the mice were fed with regular mouse chow, and the NM group, in which the mice were fed with the regular diet supplemented with 0.5% NM (w/w). After 4 weeks, thetumors were excised and processed for histology. Tumor growth was evaluated and the tumors were stained for the ECM proteins collagen I, collagen IV, fibronectin, laminin, periodic acid-Schiff (PAS) and elastin. NM strongly inhibited (by 59%, P=0.001) the growth of HeLa xenografts in nude mice. Tumors from control mice exhibited little to no collagen I expression either internally or in the fibrous capsule, while tumors from the NM group expressed collagen I in the fibrous capsule and within the tumor. Tumors from the control group showed diffuse cytoplasmic and capsular collagen IV with abundant nucleated cells. NM treatment substantially increased collagen IV production and induced a dense fibrous network of collagen IV with chambers that surrounded live nucleated cells and large amounts of necrotic cell debris. Tumors from the mice fed with the NM exhibited a well-defined border of fibronectin in the capsule and intense areas of staining internally whereas control group tumors showed less overall fibronectin with sporadic internal staining and little in the fibrous capsule. Although laminin appeared abundantly in control and NM-treated tumors, the NM group tumors exhibited a chamber-like network of laminin internally. Tumors from the control group exhibited internal areas of intense PAS staining, whereas tumors from the NM-treated group exhibited a more uniform diffuse pattern of PAS staining. In conclusion, NM supplementation of HeLa xenograft-bearing female nude mice demonstrated a potent inhibition of tumor growth and enhancement of ECM proteins, suggesting the therapeutic value of this specific nutrient complex in the treatment of cervical cancer.

Huntington's disease (HD) is a progressive neurodegenerative disorder for which only symptomatic treatments of limited effectiveness are available. Preventing early misfolding steps and thereby aggregation of the polyglutamine (polyQ)-containing protein huntingtin (htt) in neurons of patients may represent an attractive therapeutic strategy to postpone the onset and progression of HD. Here, we demonstrate that the green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) potently inhibits the aggregation of mutant htt exon 1 protein in a dose-dependent manner. Dot-blot assays and atomic force microscopy studies revealed that EGCG modulates misfolding and oligomerization of mutant htt exon 1 protein in vitro, indicating that it interferes with very early events in the aggregation process. Also, EGCG significantly reduced polyQ-mediated htt protein aggregation and cytotoxicity in an yeast model of HD. When EGCG was fed to transgenic HD flies overexpressing a pathogenic htt exon 1 protein, photoreceptor degeneration and motor function improved. These results indicate that modulators of htt exon 1 misfolding and oligomerization like EGCG are likely to reduce polyQ-mediated toxicity in vivo. Our studies may provide the basis for the development of a novel pharmacotherapy for HD and related polyQ disorders.

OBJECTIVE:Juvenile skin has been the subject of intense research efforts since ancient times. This article reports on synergistic complementarities in the biological actions of green tea and red light, which inspired the design of a green tea-assisted facial rejuvenation program.

BACKGROUND DATA:The approach is based on previous laboratory experiments providing insight into a mechanism by which visible light interacts with cells and their microenvironment.

METHODS:After 2 months of extreme oxidative stress, green tea-filled cotton pads were placed once per day for 20 minutes onto the skin before treatment with an array of light-emitting diodes (central wavelength 670 nm, dermal dose 4 J/cm2).

RESULTS:Rejuvenated skin, reduced wrinkle levels, and juvenile complexion, previously realized in 10 months of light treatment alone were realized in 1 month.

CONCLUSION:The accelerated skin rejuvenation based on the interplay of the physicochemical and biological effects of light with the reactive oxygen species scavenging capacity of green tea extends the action spectrum of phototherapy. The duo opens the gate to a multitude of possible biomedical light applications and cosmetic formulas, including reversal of topical deterioration related to excess reactive oxygen species, such as graying of hair.

OBJECTIVE:Juvenile skin has been the subject of intense research efforts since ancient times. This article reports on synergistic complementarities in the biological actions of green tea and red light, which inspired the design of a green tea-assisted facial rejuvenation program.

BACKGROUND DATA:The approach is based on previous laboratory experiments providing insight into a mechanism by which visible light interacts with cells and their microenvironment.

METHODS:After 2 months of extreme oxidative stress, green tea-filled cotton pads were placed once per day for 20 minutes onto the skin before treatment with an array of light-emitting diodes (central wavelength 670 nm, dermal dose 4 J/cm2).

RESULTS:Rejuvenated skin, reduced wrinkle levels, and juvenile complexion, previously realized in 10 months of light treatment alone were realized in 1 month.

CONCLUSION:The accelerated skin rejuvenation based on the interplay of the physicochemical and biological effects of light with the reactive oxygen species scavenging capacity of green tea extends the action spectrum of phototherapy. The duo opens the gate to a multitude of possible biomedical light applications and cosmetic formulas, including reversal of topical deterioration related to excess reactive oxygen species, such as graying of hair.

OBJECTIVE:Juvenile skin has been the subject of intense research efforts since ancient times. This article reports on synergistic complementarities in the biological actions of green tea and red light, which inspired the design of a green tea-assisted facial rejuvenation program.

BACKGROUND DATA:The approach is based on previous laboratory experiments providing insight into a mechanism by which visible light interacts with cells and their microenvironment.

METHODS:After 2 months of extreme oxidative stress, green tea-filled cotton pads were placed once per day for 20 minutes onto the skin before treatment with an array of light-emitting diodes (central wavelength 670 nm, dermal dose 4 J/cm2).

RESULTS:Rejuvenated skin, reduced wrinkle levels, and juvenile complexion, previously realized in 10 months of light treatment alone were realized in 1 month.

CONCLUSION:The accelerated skin rejuvenation based on the interplay of the physicochemical and biological effects of light with the reactive oxygen species scavenging capacity of green tea extends the action spectrum of phototherapy. The duo opens the gate to a multitude of possible biomedical light applications and cosmetic formulas, including reversal of topical deterioration related to excess reactive oxygen species, such as graying of hair.

The purpose of this study was to investigate the effect of green tea extract (GTE) supplementation combined with endurance training on endurance capacity and performance in sedentary men. Forty untrained men (age: 20± 1 years) participated in this study. Subjects were assigned to 1 of 4 treatments: (i) placebo-control (CTRL), (ii) GTE, (iii) endurance training (Ex), and (iv) endurance training with GTE (ExGTE). During the 4-week intervention, exercise training was prescribed as 75% oxygen uptake reserve for three 20-min sessions per week, and either GTE (250 mg/day) or placebo was provided. Endurance capacity, malondialdehyde (MDA), total antioxidant status (TAS), and creatine kinase (CK) were examined. Ex and ExGTE but not GTE improved exhaustive-run time (Ex: +8.2%, p = 0.031; ExGTE: +14.3%, p<0.001); in addition, Ex and ExGTE significantly increased maximal oxygen uptake by∼14% (p = 0.041) and ∼17% (p = 0.017) above the values of the CTRL group, respectively. Both Ex and ExGTE significantly decreased the increase of CK by ∼11%-32% below that of CTRL following an exhaustive run (Ex: p = 0.007; ExGTE: p = 0.001). Moreover, TAS levels increased by ∼11% in ExGTE after training (p = 0.040), and GTE, Ex, and ExGTE markedly attenuated exercise-induced MDA production (p = 0.01, p = 0.005, p = 0.011, respectively). In conclusion, this investigation demonstrated that daily ingestion of GTE during endurance training does not impair improvements in endurance capacity. Moreover, endurance training combined with GTE not only increases antioxidant capacity without attenuating endurance training adaptations, but also further attenuates acute exercise-induced CK release.

Fat oxidation has been shown to increase after short term green tea extract (GTE) ingestion and after one bout of intermittent sprinting exercise (ISE). Whether combining the two will result in greater fat oxidation after ISE is undetermined. The aim of the current study was to investigate the combined effect of short term GTE and a single session of ISE upon post-exercise fat oxidation. Fourteen women consumed three GTE or placebo capsules the day before and one capsule 90 min before a 20-min ISE cycling protocol followed by 1 h of resting recovery. Fat oxidation was calculated using indirect calorimetry. There was a significant increase in fat oxidation post-exercise compared to at rest in the placebo condition (p<0.01). After GTE ingestion, however, at rest and post-exercise, fat oxidation was significantly greater (p<0.05) than that after placebo. Plasma glycerol levels at rest and 15 min during post-exercise were significantly higher (p<0.05) after GTE consumption compared to placebo. Compared to placebo, plasma catecholamines increased significantly after GTE consumption and 20 min after ISE (p<0.05). Acute GTE ingestion significantly increased fat oxidation under resting and post-exercise conditions when compared to placebo.

BACKGROUND:Exercise and green tea supplementation have been shown to have the potential to improve postprandial blood glucose concentrations, but past interventions have not often investigated attainable and time effective exercise protocols.

AIM:The purpose of this study was to investigate the effects of interval walking exercise and acute green tea extract supplementation on the glycaemic response to an oral glucose tolerance test (OGTT).

METHOD:Twelve physically inactive participants (nine male, three female, age: 22± 1 years; body mass: 81.2 ± 16.3 kg; stature: 175.7 ± 9.6 cm; body mass index (in kg/m): 26.2± 4.3) underwent a 2-h OGTT immediately following i) no intervention (REST), ii) placebo and exercise (EX-PLAC), iii) green tea extract supplementation and exercise (EX-GTE), in a random order. The walking exercise consisted of 6 × 1 min of brisk walking (7.92 ± 0.56 km/h) separated by 1 min of slower walking (4.8 km/h). Differences between groups were identified using magnitude-based inferences.

RESULTS:The EX-GTE intervention resulted in a∼9% most likely beneficial effect on blood glucose area under the curve response to the OGTT (702.18 ± 76.90 mmol/L·120 min) compared with REST (775.30± 86.76 mmol/L·120 min), and a very likely beneficial effect compared with the EX-PLAC (772.04± 81.53 mmol/L·120 min).

CONCLUSION:These data suggest that an EX-GTE intervention can reduce postprandial glucose concentrations in physically inactive individuals.

Current treatments are generally ineffective once breast cancer has metastasized; median survival is reduced to 2-3 yr. Previous research studies demonstrating potent synergistic antitumor activity of lysine, proline, ascorbic acid, and epigallocatechin gallate prompted us to investigate the in vivo inhibitory effect of a nutrient mixture containing lysine, proline, arginine, ascorbic acid, and epigallocatechin gallate (NM) on the growth of human cancer xenografts in female athymic nude mice. Five to six week old female mice were inoculated with 3x106 breast cancer cells MDA-MB-231. After injection, the mice were randomly divided into two groups A and B; group A was fed a regular diet and group B with the regular diet supplemented with 0.5% of the nutrient mixture (NM). Four weeks later, the mice were sacrificed, and their tumors were excised, weighed, and processed for histology. We also tested the effect of NM in vitro on estrogen-receptor positive (ER+) MCF-7 and estrogen-receptor negative (ER-) MDA-MB-231 breast cancer cell lines by measuring: cell proliferation by MTT assay, expression of MMPs by gelatinase zymography, invasion through Matrigel, and VEGF by ELISA. MCF-7 cells were also treated with estradiol to study enhanced invasion and expression of MMPs and VEGF. Results showed that NM inhibited the growth and reduced the size of tumors in female nude mice by 27%. Furthermore, histological evaluation revealed increased mitotic index, MMP-9 and VEGF secretion, and PAS material (mucin) in the control group tissues. In vitro studies showed NM inhibited MDA-MB-231 cell growth by 34% at 500 microg/mL and MCF-7 cell growth by 18% at 1000 microg/mL. Invasion of MDA-MB-231 through Matrigel was inhibited by 50%, 60%, and 95% by 10, 50, and 100 microg/mL of NM, respectively. The results of this study demonstrated that the nutrient mixture tested significantly suppressed tumor growth of breast cancer cells in female athymic nude mice and significantly inhibited MMP expression, angiogenesis, and invasion in breast cancer cells, in vitro, offering promise for therapeutic use in the treatment of breast cancer.

Corneal neovascularization is a significant, sight-threatening complication of many ocular surface disorders. Various growth factors and proteinases are involved in corneal neovascularization. The data supporting a causal role for vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs) are extensive. Inhibition of VEGF and MMPs is a main strategy for treating corneal neovascularization. Several findings have shown that corneal neovascularization can be reduced by using anti-VEGF and anti-MMPs agents. Efficacy of a nutrient mixture (NM) containing lysine, proline, ascorbic acid, and green tea extract has been demonstrated for reducing VEGF and MMPs secretion by various cells. Moreover, NM can inhibit endothelial cell migration and capillary tube formation. We herein note that topical application of NM is potentially useful for inhibiting corneal neovascularization and restoration of corneal clarity. Further investigations in animal models are needed to place NM alongside corneal neovascularization therapeutics.

Degradation of extracellular matrix (ECM) is a hallmark of tumor invasion, metastasis and angiogenesis. Based on the Rath multitargeted approach to cancer using natural substances to control ECM stability and enhancing its strength, we developed a novel formulation (NM) of lysine, proline, ascorbic acid and green tea extract that has shown significant anti-cancer activity against a number of cancer cell lines. The aim of the present study was to determine whether NM exhibits anti-angiogenic and anti-metastatic effects using in vitro and in vivo experimental models. Angiogenesis was measured using a chorioallantoic membrane (CAM) assay in chick embryos and bFGF-induced vessel growth in C57BL/6J female mice. To determine the in vivo effect of NM on the tumor xenograft growth, male nude mice were inoculated with 3 x 10(6) MNNG-HOS cells. Control mice were fed a mouse chow diet, while the test group was fed a mouse chow diet supplemented with 0.5% NM for 4 weeks. In vitro studies on cell proliferation (MTT assay), MMP expression (zymography) and Matrigel invasion were conducted on human osteosarcoma U2OS, maintained in McCoy medium, supplemented with 10% FBS, penicillin and streptomycin in 24-well tissue culture plates and tested with NM at 0, 10, 50, 100, 500, and 1000 microg/ml in triplicate at each dose. NM at 250 microg/ml caused a significant (p<0.05) reduction in bFGF-induced angiogenesis in CAM. NM inhibited tumor growth of osteosarcoma MNNG-HOS cell xenografts in nude mice by 53%; furthermore, tumors in NM-treated mice were less vascular and expressed lower levels of VEGF and MMP-9 immunohistochemically than tumors in the control group. In addition, NM exhibited a dose-dependent inhibition of osteosarcoma U2OS cell proliferation (up to 60% at 1000 microg/ml), MMP-2 and -9 expression (with virtual total inhibition at 500 microg/ml NM), and invasion through Matrigel (with total inhibition at 100 microg/ml NM). Moreover, NM decreased U2OS cell expression of VEGF, angiopoietin-2, bFGF, PDGF and TGFbeta-1. These results together with our earlier findings suggest that NM is a relatively non-toxic formulation, which inhibits growth, invasion, metastasis, and angiogenesis of tumor cells.