Matrix metalloproteinase (MMP)-2 and -9 secretion is elevated in various human cancers and their elevated expression has been associated with poor prognosis due to associated increased cancer cell ...invasion and metastasis.
To examine the correlation between in vitro MMP-2 and MMP-9 secretion and Matrigel invasion in 42 different human cancer cell lines (selected on the basis of organ malignancies) treated with a nutrient mixture (NM).
The cells were cultured in their recommended media supplemented with 10% FBS and antibiotics in 24-well tissue culture plates. At near confluence, the cells were treated with NM dissolved in media at 0, 10, 50, 100, 500 and 1000 μg/mL in triplicate. Parallel sets of cultures were also treated with phorbol 12-myristate 13-acetate (PMA) 100 ng/mL for induction of enzymes. After 24 h the media were collected and MMP-2 and MMP-9 levels were assayed by gelatinase zymography. Invasion studies were conducted using Matrigel in 24-well plates.
Correlation of pooled data from different cancer cell line groups demonstrated dose-dependent inhibition of MMP-2 and -9 and Matrigel invasion with NM treatment and significant negative correlation between MMP-2 and MMP-9 levels and Matrigel invasion. Pooled data of cell lines expressing only MMP-2 and resistance to PMA induction of MMP-9 showed significant negative correlation (r = -0.77, p = 0.003) between MMP-2 secretion and inhibition of invasion through Matrigel. Cell lines expressing only MMP-9, showed significant negative correlation (r = -0.726, p = 0.003) between MMP-9 secretion and Matrigel invasion. Pooled data of cell lines expressing MMP-2 and MMP-9 demonstrated significant negative correlation (r = -0.821, p < 0.0001) between MMP-9 secretion and inhibition of invasion through Matrigel. Pooled data of cancer cell lines expressing no basal MMP- 9 secretion demonstrated significant negative correlation (r = -0.686, p < 0.0001) between PMA-induced MMP-9 secretion and inhibition of invasion through Matrigel.
In conclusion, regardless of MMP-2 and MMP-9 patterns of expression, MMP modulation by NM was found to be significantly correlated with NM modulation of Matrigel invasion of these cell lines.
Acetaminophen (APAP) overdose is often fatal, leading to fulminant hepatic and renal tubular necrosis in humans and animals. We studied the effect of a nutrient mixture (NM) containing, among other ...nutrients, lysine, proline, ascorbic acid, N-acetyl cysteine, and green tea extract, which has previously been demonstrated to exhibit a broad spectrum of therapeutic properties on APAP-induced hepatic and renal damage in ICR (Imprinting Control Region) mice. Seven-week-old male ICR mice were divided into four groups (A–D) of five animals each. Groups A and C mice were fed a regular diet for 2 weeks, while groups B and D mice were supplemented with 0.5% NM (w/w) during that period. Groups A and B received saline i.p., while groups C and D received APAP (600 mg/kg) i.p. All animals were killed 24 h after APAP administration, serum was collected to assess the liver and kidney functions, and the livers and kidneys were excised for histology. Mean serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, BUN (Blood Urea Nitrogen), creatinine, and BUN/creatinine ratios were comparable in groups A and B, increased markedly in group C and significantly lower in group D compared with group C. APAP caused significant centrilobular necrosis and glomerular damage in unsupplemented animals, while NM prevented these alterations. The results indicate that NM has potential to protect against APAP-induced liver and kidney damage.
Autologous blood patch pleurodesis (ABPP) offers an alternative to surgery when conservative management is ineffective for treating a persistent air leak (PAL). In the traditional technique, ...autologous venous blood is introduced into the affected pleural cavity via a blindly inserted large-bore surgical chest tube. Herein, we present a case of an 18 year old male with a PAL following video assisted thorascopic bleb resection and talc pleurodesis who underwent successful ABPP using a small-bore pigtail catheter placed under computed tomography (CT) guidance. As compared to the traditional technique, this approach may potentially offer several advantages, such as more precise chest tube placement over the air leak site and reduced pain with chest tube insertion. Although image-guided chest tube insertion is a commonly performed procedure, its use for the specific indication in this patient's case has not been described in the literature, to the best of our knowledge.
•Persistent air leak (PAL) is a commonly encountered complication following thoracic surgery.•Autologous blood patch pleurodesis (ABPP) may be performed to treat a PAL if conservative management is unsuccessful.•Traditionally, ABPP involves instilling venous blood into the pleural space via a blindly inserted large-bore chest tube.•We report a case in which ABPP was performed using a small-bore catheter placed under Computed Tomography (CT) guidance.•The use of CT-guided small-bore catheter insertion may offer several advantages as compared to the traditional technique.
Fanconi Anemia, an autosomal recessive disorder, is characterized by chromosomal abnormality leading to birth defects, progressive bone marrow failure, and a high probability of developing malignancy ...at an early age. Head and neck squamous cell carcinoma and myeloid leukemia are the major causes of cancer related morbidity and mortality in Fanconi anemia patients.
We investigated the effect of a nutrient mixture on Fanconi Anemia human fibroblast cell lines FA-A:PD20 and FA-A:PD220 on matrix metalloproteinase expression, invasion, cell proliferation, morphology and apoptosis. The cell lines were grown in a modified Dulbecco's Eagle medium and at near confluence were treated with the nutrient mixture at increasing doses: 0; 10; 50; 100; 500; 1000 µg/ml. The cells were also treated with PMA to induce MMP-9 expression.
Zymography demonstrated MMP-2 and PMA-induced MMP-9 activity. The nutrient mixture inhibited expression of both, MMP-2 and MMP-9, in a dose dependent manner with virtually total inhibition observed at 500 µg/ml. Matrigel invasion was inhibited in both cells lines; with 100% inhibition for FA-A:PD20 at 500 µg/ml and 100% inhibition of FA-A:P220 cells at 100 µg/ml. H&E staining did not indicate any change in cell morphology and causes apoptosis at higher doses.
Our data demonstrated that the nutrient mixture inhibited matrix metalloproteinase expression, invasion and induced apoptosis, the important parameters for cancer prevention. The results suggest that the nutrient mixture may have therapeutic potential in Fanconi Anemia associated neoplasia.
A specific nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract has demonstrated a broad spectrum of antitumor activity against a number of cancer cell lines. In this ...study, our main objective was to investigate the comparative effects of NM on anticancer parameters, such as cytotoxicity, matrix metalloproteinase (MMP) secretion and Matrigel invasion in the human uterine sarcoma drug-resistant MES-SA/Dx5 and the drug-sensitive MES-SA cell lines. In addition we studied the effects of NM on P-glycoprotein (Pgp) on these cell lines. Cell proliferation was evaluated by MTT assay, MMPs by gelatinase zymography, invasion through Matrigel, morphology by H&E and Pgp expression by Western blot analysis and immunodetection using FITC-conjugated antibody and rhodamine 123 (Rh123) accumulation and efflux assays. NM exhibited antiproliferative effects on MES-SA/Dx5, by 20% at 50 and 100 µg/ml and by 36, 40 and 48% at 250, 500 and 1,000 µg/ml, respectively. By contrast, NM treatment of MES-SA cells resulted in significantly increased cytotoxicity: 40, 46, 65 and 72% at 50, 100, 500 and 1,000 µg/ml, respectively. In both cell lines, zymography demonstrated a band corresponding to MMP-2 in normal cells and MMP-9 with phorbol 12-myristate 13-acetate treatment. The two MMPs showed dose-response inhibition by NM. As shown by Western blot analysis and immunodetection, NM treatment resulted in a dose-dependent decrease in Pgp expression in the MES-SA/Dx5 cell line. The MES-SA cell line does not exhibit Pgp. NM enhanced the accumulation and efflux of the Pgp substrate, Rh123, in the MES-SA/Dx5 uterine sarcoma cell line but not in the drug-sensitive cell line, MES-SA. Therefore, it can be concluded that NM demonstrates potent anticancer effects in both the drug-resistant and sensitive cell lines and modulates Pgp, suggesting its potential therapeutic effects in drug-resistant as well as sensitive cancers.
Context. The extremely relativistic Fe lines, detected from some active galactic nuclei (AGN), indicate that generation and reprocessing of the X-ray emission takes place in the immediate vicinity of ...the event horizon. Recently, general relativistic (GR) effects, in particular light bending which is very strong in that region, have been considered to be the cause of complex variability patterns observed in these AGNs. Aims. We study in detail the GR models of the X-ray spectral variability for various geometries of the X-ray source and with various relativistic effects being the dominant cause of spectral variability. The predicted properties are compared with the observational data of the Seyfert 1 galaxy MCG–6-30-15, which is currently the best studied AGN with signatures of strong gravity effects. Methods. We focus on modeling the root mean square (rms) spectra. We compute the rms spectra for the GR models using a Monte Carlo method, and compare them with the rms spectra from the Suzaku observations of MCG–6-30-15 on January 2006. Results. The data disfavor models with the X-ray source (1) moving vertically on the symmetry axis or (2) corotating with the disc and changing height not far above the disc surface. The most likely explanation for the observed fractional variability is given by the model involving the X-ray source located at a very small, varying distance from a rapidly rotating black hole. This model predicts some enhanced variations in the red wing of the Fe line, which are not seen in the Suzaku observations. However, the enhanced variability of the red wing, while ruled out by the Suzaku data, is consistent with an excess rms variability, between 5 and 6 keV, reported for some previous ASCA and XMM observations. We speculate that the presence or lack of such a feature is related to the change of the ionization state of the innermost part of the disc, however, investigation of such effects is currently not possible in our model (where a neutral disc is assumed). If the model, completed by a description of ionization effects, proves to be fully consistent with the observational data, it will provide a strong indication that the central black hole in MCG–6-30-15 rotates rapidly, supporting similar conclusions derived from the Fe line profile.
A nutrient mixture (NM) containing ascorbic acid, lysine, proline and green tea extract has exhibited anticancer activity in vitro and in vivo in a number of cancer cell lines. We investigated the ...effect of NM on human leukemic myeloid U-937 cells in vitro by measuring: cell proliferation, MMP expression, invasion, apoptosis, and COX-2 and COX-1 protein expression.
Human leukemic cell line U-937 (ATCC) was cultured in RPMI medium supplemented with fetal bovine serum and antibiotics. After 24 h, the cells were treated with NM at 0, 50, 100, 250, 500 and 1000 Ојg/ml, in triplicate at each dose. Phorbol 12-myristate 13-acetate (PMA), 100 ng/ml was added to cells to induce MMP-9 secretion. Cell proliferation was evaluated by MTT assay, MMP expression by gelatinase zymography, invasion through Matrigel, apoptosis by using live green caspase detection kit (Molecular Probe), and COX-2 and COX-1 expression by Western blot.
NM had no effect on U-937 cell growth at a concentration of 250 Ојg/ml and exhibited an antiproliferative effect at 500 Ојg/ml concentration. Zymography did not demonstrate MMP-2 or MMP-9 secretion in normal cells; however, PMA strongly induced MMP-9, which was inhibited by NM in a dose-dependent manner. Cell penetration through Matrigel was significantly reduced (by 95%) at 250 Ојg/ml NM and completely blocked at 500 Ојg/ml NM. NM induced slight apoptosis at 100 Ојg/ml and moderate at 500 and 1000 Ојg/ml concentration. NM inhibited COX-2 expression in a dose-dependent fashion and had no effect on COX-1 expression.
Our results suggest that NM has potent inhibitory effects on U-937 cell growth and expression of inflammatory mediators, significant parameters in AML progression.
influenza, one of the oldeist and most common infections, poses a serious health problem causing significant morbidity and mortality, and imposing substantial economic costs. The efficacy of current ...drugs is limited and improved therapies are needed. A unique nutrient mixture (NM), containing ascorbic acid, green tea extract, lysine, proline, N‐acetyl cysteine, selenium among other micronutrients, has been shown to exert anti‐carcinogenic and anti‐atherogenic activity both in vitro and in vivo. Many of the constituents of NM have been shown to have an inhibitory effect on replication of influenza virus and HIV. This prompted us to study the effect of NM on influenza A virus multiplication in infected cells and neuraminidase activity (NA) in virus particles. Addition of NM to Vero or MDCK cells post infection resulted in dose‐dependent inhibition of viral nucleoprotein (NP) production in infected cells. NM‐mediated inhibition of viral NP was selective and not due to cytotoxicity towards host cells. This antiviral effect was enhanced by pretreatment of virus with the nutrient mixture. Individual components of NM, namely ascorbic acid and green tea extract, also blocked viral NP production, conferring enhanced inhibition when tested in combination. Incubation of cell‐free virus with NM resulted in dose‐dependent inhibition of associated NA enzyme activity. In conclusion, the nutrient mixture exerts an antiviral effect against influenza A virus by lowering viral protein production in infected cells and diminishing viral enzymatic activity in cell‐free particles.
Long-term survival of patients with hepatocellular carcinoma (HCC), a common cancer worldwide, remains poor, due to metastasis and recurrence.
To investigate the effect of a novel nutrient mixture ...(NM) containing ascorbic acid, lysine, proline, and green tea extract on human HCC cell line Sk-Hep-1 In vivo and In vitro.
After one week of isolation, 5-6 week old male athymic nude mice were inoculated with 3 x 10(6) SK-Hep-1 cells subcutaneously and randomly divided into two groups; group A was fed a regular diet and group B a regular diet supplemented with 0.5% 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 SK-Hep-1 cells, measuring cell proliferation by MTT assay, invasion through Matrigel, apoptosis by green caspase detection kit, MMP secretion by zymography, and morphology by H&E staining.
NM inhibited tumor weight and burden of SK-Hep-1 xenografts by 42% and 33% respectively. In vitro , NM exhibited 33% toxicity over the control at 500 and 1,000 microg/ml concentration. Zymography demonstrated MMP-2 and MMP-9 secretion which was inhibited by NM in a dose dependent fashion, with virtual total inhibition at 1000 microg/ml. Invasion through Matrigel was inhibited at 100, 500 and 1,000 microg/ml by 53%, 83% and 100% respectively. NM induced slight apoptosis at 100 microg/ml, and profound apoptosis at 500 microg/ml and 1000 microg/ml concentration.
These results suggest that NM has therapeutic potential in treatment of HCC.
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