Ramtej J. Verma

PROGNOSTIC SIGNIFICANCE OF MOLECULAR MARKERS IN ORAL SQUAMOUS CELL CARCINOMA: A MULTIVARIATE ANALYSIS

Multiple marker accumulation impacts tumor progression and biologic phenotypes affect clinical outcome of patients with head and neck cancer. Hence, this study investigated a battery of molecular markers that may help to reflect biologic aggressiveness and predict prognosis.

Isolation, characterization and antioxidative effect of phyllanthin against CCl4-induced toxicity in HepG2 cell line

The present study was an attempt to investigate the hepatoprotective and antioxidative property of Phyllanthus amarus (P. amarus) extract and phyllanthin. Phyllanthin, one of the active lignin present in this plant species was isolated from the aerial parts, by silica gel column chromatography employing gradient elution with hexane-ethyl acetate solvent mixture. It was obtained in high yields (1.23%), compared to reported procedures and the purity was ascertained by HPTLC and reversed-phase HPLC analysis. Characterization of phyllanthin was done by mp, UV-Visible spectrophotometry, elemental analysis, FT-IR, 1H NMR, 13C NMR and mass spectral analysis. Free radical scavenging activity of P. amarus extract and phyllanthin was also examined using DPPH assay. The protective effect of P. amarus extract and phyllanthin was studied on CCl4-induced toxicity in human hepatoma HepG2 cell line. The results indicated that CCl4 treatment caused a significant decrease in cell viability. In addition, the toxin treatment initiated lipid peroxidation (LPO), caused leakage of enzymes like alanine transaminase (ALT) and lactate dehydrogenase (LDH) with a significant decrease in glutathione (GSH) levels. It was observed that phyllanthin effectively alleviated the changes induced by CCl4 in a concentration-dependent manner, with much smaller strengths as compared to P. amarus extract.

Molecular alterations in oral carcinogenesis: significant risk predictors in malignant transformation and tumor progression.

In this study an attempt was made to establish the significance of a battery of molecular alterations and thereby identify risk predictors in oral carcinogenesis. For this purpose, EGFR, Stat3, H-ras, c-myc, p53, cyclin D1, p16, Rb, Ki-67 and Bcl-2 were localized immunohistochemically in normal mucosa (n=12), hyperplasia (n=35), dysplasia (n=25), early stage carcinoma (n=65) and advanced stage carcinoma (n=70). Deregulation occurred at an early stage and the number of alterations increased with disease progression. Using multivariate logistic regression analysis, the significant risk predictor for hyperplasia from normal mucosa was Ki-67 (OR=5.75, p=0.021); the significant risk predictors for dysplasia from hyperplasia were EGFR (OR=12.96, p=0.002), Stat3 (OR=17.16, p=0.0001), p16 (OR=5.50, p=0.039) and c-myc (OR=5.99, p=0.052); the significant risk predictors for early stage carcinoma from dysplasia were p53 (OR=6.63, p=0.0001) and Rb (OR=3.81, p=0.056); and the significant risk predictors for further progression were EGFR (OR=5.50, p=0.0001), Stat3 (OR=4.49, p=0.0001), H-ras (OR=4.05, p=0.001) and c-myc (OR=2.99, p=0.015). Cyclin D1 holds a key position linking upstream signaling pathways to cell cycle regulation. Gene products of the mitogenic signaling pathway play an equally significant role as cell cycle regulatory proteins in the hyperplasia-dysplasia-early-advanced-carcinoma sequence and together may provide a reference panel of markers for use in defining premalignant lesions and predicting the risk of malignant transformation and tumor progression.

Curcumin ameliorates aflatoxin-induced toxicity in mice spermatozoa

OBJECTIVE To investigate the ameliorative effect of curcumin on aflatoxin-induced toxicity in mice spermatozoa. DESIGN Prospective controlled animal study. SETTING University research laboratory. PATIENT(S) Young, inbred, Swiss-strain male albino mice (Mus musculus), weighing approximately 37 to 40 g, were obtained from Cadila Health Care, Ahmedabad, India. INTERVENTION(S) Aflatoxin was orally administered in 25 (low dose) and 50 (high dose) microg/0.2 mL olive oil to each animal, each day (750 and 1500 microg/kg body weight), respectively, with and without curcumin for 45 days. On 46th day the animals were killed by cervical dislocation. The cauda epididymis were removed and weighed, then was teased in normal saline to obtain spermatozoa. MAIN OUTCOME MEASURE(S) Sperm count, viability, and motility were assessed immediately. Sperm semidried smears were prepared on slides and stained by Papanicolaou stain to study sperm morphologic features. RESULT(S) There was a dose-dependent spermicidal effect of aflatoxin. Sperm count, viability, and motility were statistically significantly reduced. Different morphologic abnormalities were encountered. Treatment with curcumin along with aflatoxin ameliorated aflatoxin-induced sperm count, immobilization, and viability, and improved the morphologic characteristics of the sperm. CONCLUSION(S) Curcumin has an ameliorative effect on sperm parameters and improves morphologic features of sperm in mice.

Sodium fluoride-induced hypoproteinemia and hypoglycemia in parental and F1-generation rats and amelioration by vitamins

Oral administration of sodium fluoride (NaF; 40 mg/kg body weight) daily from day 6 of gestation to day 21 of lactation caused, compared with the distilled water control (group 2), significant reductions in body weight and feed consumption as well as concentration of glucose and protein in the serum of P- and F(1)-generation rats; however, sodium and potassium concentrations in the serum were significantly higher than those of the vehicle control (group 2). Administration of either vitamins C (50 mg/kg body weight/day), D (2 ng/0.2 ml olive oil/animal/day) or a combination of vitamins C+D+E along with NaF caused significant amelioration in body weight and feed consumption, as well as glucose, protein, sodium and potassium concentrations in the serum of P- and F(1)-generation rats compared with the NaF-only treated group. Withdrawal of NaF treatment during lactation caused significant amelioration in feed consumption (days 15-21 only), sodium, potassium, glucose and protein concentrations in the serum of both P- and F(1)-generation rats. Co- treatment with vitamin E (2 mg/0.2 ml olive oil/animal/day) caused significant amelioration in body weight (days 15 and 20 of gestation only), sodium, potassium, glucose (only in P-generation females) and protein (only in P-generation female) concentrations in the serum of rats than in NaF-treated rats alone. It is concluded that co-treatment with vitamins C, D and C+D+E were found more effective in ameliorating NaF-induced effects than vitamin E and withdrawal of NaF treatment during lactation.

Effect of aflatoxins on testicular steroidogenesis and amelioration by vitamin E

The potential of aflatoxin to affect testicular steroidogenesis and its amelioration by vitamin E was assessed in the present investigation. Oral administration of aflatoxin (25 and 50 microg/animal/day) for 45 days to adult mice caused, as compared with control, a dose-dependent significant rise in cholesterol content. However, the activities of 3beta- and 17beta-hydroxysteroid dehydrogenases and serum testosterone levels were significantly reduced in aflatoxin-treated mice as compared with the controls. Pretreatment with vitamin E (2 mg/animal/day, orally) significantly ameliorated aflatoxin-induced changes as compared with aflatoxin treatment alone. The present investigation clearly indicates that vitamin E ameliorates aflatoxin-induced changes in steroidogenesis.

Stat3 expression in oral squamous cell carcinoma: association with clinicopathological parameters and survival

The present study sought to explore the occurrence of signal transducer and activator of transcription 3 (Stat3) in patients with oral squamous cell carcinoma (n=135) and its potential relationship with clinicopathological parameters and survival. Stat3 expression was studied by immunohistochemistry. Cytoplasmic or nuclear localization of Stat3 was observed in 62% of patients, whereas only nuclear Stat3 expression was found in 44%. Stat3 positivity in early-stage patients was 45% compared to 79% in advanced-stage patients. However, early-stage Stat3-positive patients showed a gradual increase in staining intensity, with intense staining seen in 52% of the tumors compared to 18% in Stat3-positive advanced-stage patients, where a gradual decrease in intensity expression was observed (p=0.001). Stat3 showed a significant positive correlation with disease stage (p=0.001), nodal status (p=0.033) and tumor size (p=0.001). Multivariate survival analysis using the Cox proportional hazard regression model showed that nuclear Stat3 was a significant independent prognosticator for both relapse-free survival (p=0.014) and overall survival (p=0.042) in early-stage patients. Our results indicated that Stat3 activation is an early event in oral squamous cell carcinoma and represents a potential risk factor for poor prognosis in early-stage patients.

Cytotoxicity of aflatoxin on red blood corpuscles

The exact mechanism of aflatoxin action is not clearly understood. In the present investigation the authors report morphological aberrations and increased rate of hemolysis caused by aflatoxins in vitro.

Effect of curcumin on aflatoxin-induced biochemical changes in testis of mice.

OBJECTIVE To investigate the effect of curcumin on aflatoxin-induced biochemical changes in testis of mice. DESIGN Controlled research laboratory study. SETTING Aflatoxin was obtained by growing Aspergillus parasiticus in SMKY liquid medium. Pure curcumin (97% purity) was purchased from Hi-Media Laboratories Pvt. Ltd., Mumbai, India. PATIENT(S) Young inbred, Swiss strain male albino mice (Mus musculus), weighing approximately 37-40 g, were obtained from Cadila Health Care, Ahmedabad, India. INTERVENTION(S) Aflatoxin was administered orally at 25 microg (low dose) and 50 microg (high dose)/0.2 mL olive oil/animal/day (750 and 1,500 microg/kg body weight), respectively, with and without curcumin for 45 days. On the 46th day the animals were sacrificed by cervical dislocation. Testis were removed and weighed. Homogenates were prepared for respective parameters such as protein, cholesterol, DNA, RNA, and 3beta- and 17beta-hydroxysteroid dehydrogenase activity. MAIN OUTCOME MEASURE(S) Protein, cholesterol, DNA, RNA, 3beta- and 17beta-hydroxysteroid dehydrogenase activity were measured immediately. RESULT(S) There was a dose-dependent effect of aflatoxin on testis of mice. Protein, cholesterol, DNA, RNA, and 3beta- and 17beta-hydroxysteroid dehydrogenase activity were reduced significantly by the treatment of aflatoxin; cholesterol was increased. Treatment with curcumin along with aflatoxin ameliorates aflatoxin-induced biochemical changes in the testis of mice. CONCLUSION(S) Curcumin significantly ameliorates aflatoxin-induced biochemical changes in the testis of mice.

Ameliorative effect of Emblica officinalis aqueous extract on ochratoxin-induced lipid peroxidation in the kidney and liver of mice.

OBJECTIVES The present study was an attempt at investigating whether the aqueous extract of Emblica officinalis may have an ameliorative effect on ochratoxin-induced lipid peroxidation in the kidney and liver of mice. METHODS Adult male albino mice were orally administered 50 microg (LD, low dose) and 100 microg (HD, high dose) of ochratoxin/0.2 ml of olive oil/animal/day for 45 days. RESULTS The results revealed a significant (p < 0.05), dose-dependent increase in lipid peroxidation as well as a decreased activity of enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione transferase) and non-enzymatic antioxidants (glutathione and total ascorbic acid) in both the organs, as compared to the findings for olive oil-treated control group. Administration of Emblica officinalis aqueous extract (2 mg/animal/day) and ochratoxin for a period of 45 days caused a significant amelioration in the ochratoxin-induced lipid peroxidation in mouse liver and kidney.