A Randomized, Double-Blind, Placebo-Controlled Trial Evaluating the Effect of Topical Urea for Secondary Prophylaxis of Hand Foot Skin Reaction in Renal Cell Cancer Patients on Sunitinib Therapy.

INTRODUCTION: Hand foot skin reaction (HFSR) is a common dose-limiting adverse effect of multi kinase inhibitors (MKI) whose mechanism is not fully understood, and the prophylaxis is inadequate. OBJECTIVE: In this pilot study, a double-blind, randomized placebo-controlled trial was conducted to evaluate the effect of topical urea in secondary prevention of sunitinib-induced HFSR in renal cell cancer patients. METHODS: Out of 55 screened patients, 14 were randomized to receive topical urea or placebo for four weeks. The association of HFSR with drug levels of sunitinib and its metabolite (n-desethyl sunitinib), genetic polymorphism of VEGFR2 gene, quality of life (QOL) and biochemical markers was also assessed. RESULTS: The results showed that urea-based cream was not superior to placebo (P = .075). There was no change in the QOL in both the groups. Single nucleotide polymorphism was checked for two nucleotides rs1870377 and rs2305948 located in VEGFR2 gene on chromosome 4. SNP (variant T > A) at rs1870377 was associated with appearance of new HFSR as compared to the wild type, although the association was not statistically significant (OR 0.714). There was no statistically significant difference between mean plasma levels of sunitinib and N-desethyl sunitinib in urea arm as compared to placebo arm as compared to placebo. The best fit population pharmacokinetic model for sunitinib was one compartment model with first order absorption and linear elimination. The median (IQR) of population parameters calculated from the population pharmacokinetics model for Ka, V and Cl was 0.22 (0.21-0.24) h-1, 4.4 (4.09-4.47) L, 0.049 (0.042-0.12) L/hr, respectively. CONCLUSION: The study suggested that the urea-based cream was not superior to placebo in decreasing the appearance of new HFSR in renal cancer patients receiving 4:2 regimen of sunitinib.

Chemoprevention with aqueous extract of Butea monosperma flowers results in normalization of nuclear morphometry and inhibition of a proliferation marker in liver tumors.

Butea monosperma (Lam.) (family: Fabaceae) popularly known as 'Palas' or 'fire of forest' has been used traditionally as a hepatoprotective agent. This study evaluated the hepatoprotective and antitumorigenic properties of the aqueous extract and butanol fractions of B. monosperma flowers in animal models. Dried flowers of B. monosperma were extracted with water and fractionated further using n-butanol. The hepatoprotective activity of the aqueous extract was initially confirmed in a carbon tetrachloride-induced liver damage model of rats. Oral administration of the aqueous extract produced a strong hepatoprotective effect similar to silymarin and normalized the serum levels of ALT, AST, bilirubin and triglyceride in rats. However, it did not affect the levels of glutathione and malondialdehyde which are oxidative stress markers in liver. Intraperitoneal administration of the aqueous extract in the X15-myc oncomice not only maintained liver architecture and nuclear morphometry but also down-regulated the serum VEGF levels. Immunohistochemical staining of liver sections with anti-Ribosomal protein S27a antibody showed post-treatment abolition of this proliferation marker from the tumor tissue. The butanol fractions, however, did not show antitumorigenic activity. Thus, the aqueous extract of B. monosperma flowers is not only hepatoprotective but also antitumorigenic by preserving the nuclear morphometry of the liver.

Atorvastatin ameliorates inflammatory hyperalgesia in rat model of monoarticular arthritis.

Statins, the cholesterol lowering drugs, have been shown to exhibit anti-inflammatory properties. The aim of the present study was to evaluate the efficacy of atorvastatin in ameliorating joint dysfunction associated with arthritis. Monoarticular arthritis was induced by the intra-articular injection of FCA (0.1mL of 0.1%). The effect of atorvastatin (10 and 50mg/kg, A10 and A50) following oral administration was evaluated on joint inflammation, locomotor function and hyperalgesia daily for first 4 days and every 4th day till 28 days. The effect of atorvastatin was compared with that of diclofenac (5mg/kg, D5). Daily oral administration of atorvastatin produced a significant reduction in joint inflammation (21% in A10 and 33% in A50) and associated hyperalgesia. Atorvastatin also produced a marked improvement in the stair climbing ability and motility of the arthritic rats. The beneficial effect of atorvastatin was also evident from the histological analysis of joint carried out on day 28. Our results show that atorvastatin is more effective in decreasing the joint inflammation and hyperalgesia as compared to diclofenac while the efficacy of both the drugs in ameliorating functional disability was comparable.

Atorvastatin exhibits anti-inflammatory and anti-oxidant properties in adjuvant-induced monoarthritis.

Arthritis is a joint disorder where the joint damage is associated with elevated levels of inflammatory mediators and reactive oxygen species (ROS). The inflammatory hyperalgesia associated with arthritis has been shown to be attenuated by anti-hyperlipidemic drug, atorvastatin. The present study was carried out to evaluate the effect of atorvastatin on joint inflammation and associated oxidative stress markers in a rat model where arthritis was induced by intra-articular injection of 0.1 ml of 0.1% Freund's Complete Adjuvant (FCA). Atorvastatin (10 mg and 50 mg/kg) and diclofenac (5 mg/kg) were administered orally, daily during the study period of 4 days and their effect on joint inflammation was evaluated by measuring joint diameter, levels of glutathione (GSH), thiobarbituric acid reactive substances (TBARS), activity of super oxide dismutase (SOD) and tissue histology. Atorvastatin produced a dose-dependent reduction in joint inflammation that was associated with normalization of levels of oxidative stress markers and tissue histology and its effect was found to be comparable to that of diclofenac.

Protective effect of latex of Calotropis procera in Freund's Complete Adjuvant induced monoarthritis.

The protective effect of latex of Calotropis procera in Freund's Complete Adjuvant (FCA) induced monoarticular arthritis was evaluated in rats. Arthritis was induced by a single intra-articular injection of 0.1 mL of 0.1% FCA in the right ankle joint. The effect of dried latex (DL, 200 and 400 mg/kg) and its methanol extract (MeDL, 50 and 500 mg/kg) following oral administration was evaluated on joint inflammation, hyperalgesia, locomotor function and histology at the time of peak inflammation. The effects of DL and MeDL were compared with antiinflammatory drugs phenylbutazone (100 mg/kg), prednisolone (20 mg/kg), rofecoxib (20 and 100 mg/kg) and immuno-suppressant methotrexate (0.3 mg/kg). Daily oral administration of DL and its methanol extract (MeDL) produced a significant reduction in joint inflammation (about 50% and 80% inhibition) and associated hyperalgesia. The antihyperalgesic effect of MeDL was comparable to that of rofecoxib. Both DL and MeDL produced a marked improvement in the motility and stair climbing ability of the rats. The histological analysis of the arthritic joint also revealed significant reduction in oedema and cellular infiltration by MeDL that was comparable to that of rofecoxib. Thus, our study suggests that the latex of C. procera has the potential to be used as an antiarthritic agent.

The effect of casein phosphopeptide-amorphous calcium phosphate on remineralization of artificial caries-like lesions: an in vitro study.

BACKGROUND: The aims of this study were to investigate the efficacy of CPP-ACP containing Tooth Mousse on the remineralization of enamel lesions and to compare its efficacy to that of a fluoride-containing toothpaste. METHODS: Permanent teeth were placed in demineralizing solution for 96 hours to produce artificial caries-like lesions 120-200 microm in depth. They were sectioned into 100-150 microm thick samples and randomly assigned to five groups: for Group A, a fluoridated toothpaste (1100 ppm) was used as a positive control and in Group B, a non-fluoridated toothpaste was used as a negative control. Tooth Mousse containing CPP-ACP was tested by three different means: as a toothpaste (Group C); as a topical coating (Group D); and (Group E) as a topical coating after treating the sections with the same fluoridated toothpaste as in Group A. RESULTS: The lesion depth decreased significantly by 7 per cent in Group A, 10.1 per cent in Groups C and D, and 13.1 per cent in Group E (Paired t- test, p < 0.05), while in Group B the lesion depth increased significantly by 23 per cent. CONCLUSIONS: Based on the data obtained, CPP-ACP containing Tooth Mousse remineralized initial enamel lesions and it showed a higher remineralizing potential when applied as a topical coating after the use of a fluoridated toothpaste.

Calotropis procera latex affords protection against carbon tetrachloride induced hepatotoxicity in rats.

In the present study, latex of Calotropis procera possessing potent antioxidant and anti-inflammatory properties was evaluated for its hepatoprotective effect against carbon tetrachloride (CCl(4)) induced hepatotoxicity in rats. Subcutaneous injection of CCl(4,) administered twice a week, produced a marked elevation in the serum levels of aspartate transaminase (AST), alanine transaminase (ALT) and tumor necrosis factor alpha (TNF-alpha). Histological analysis of the liver of these rats revealed marked necro-inflammatory changes that were associated with increase in the levels of TBARS, PGE(2) and catalase and decrease in the levels of glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx). Daily oral administration of aqueous suspension of dried latex (DL) of Calotropis procera at 5, 50 and 100mg/kg doses produced a dose-dependent reduction in the serum levels of liver enzymes and inflammatory mediators and attenuated the necro-inflammatory changes in the liver. The DL treatment also normalized various biochemical parameters of oxidative stress. Our study shows that the antioxidant and anti-inflammatory effects of DL and silymarin were comparable and suggests that DL could be used as a hepatoprotective agent.

Evaluation of cytotoxic potential of latex of Calotropis procera and podophyllotoxin in Allium cepa root model.

In the present study we have utilized the Allium cepa root tip meristem model to evaluate the cytotoxic and anti-mitotic activities of latex of Calotropis procera (DL) and podophyllotoxin. Standard cytotoxic drug cyclophosphamide and non-cytotoxic drugs cyprohcptadine and aspirin served as controls. Like cyclophosphamide, both DL and podophyllotoxin significantly inhibited the growth of roots and mitotic activity in a dose-dependent manner. However, podophyllotoxin was more potent in this regard and produced root decay. Cyproheptadine and aspirin, on the other hand, showed a marginal effect on the root growth and mitotic activity at much higher concentrations.

In vivo functional analysis of the mouse estrogen receptor gene promoter: a transgenic mouse model to study tissue-specific and developmental regulation of estrogen receptor gene transcription.

Understanding the molecular and morphological basis of estrogen responsiveness in the various tissues and organs that make up an adult organism and its onset during ontogenesis requires identification of the genetic controls that determine timed expression of the estrogen receptor (ER) gene in multiple cell types. With this goal in mind, we describe here the results of the functional analysis of the mouse (m) ER gene promoter, carried out in vivo in transgenic mice. The mER gene promoter was cloned and spliced to the coding sequence of the bacterial lacZ gene (fused to the nuclear localization signal of SV40 large T: nls-beta-GAL) and then stably reintegrated into the genome of mice. Analysis of beta-GAL mRNA and protein expression in multiple organs of both female and male transgenic animals was then performed. Results show that the transgenic mER promoter, much like the endogenous one, is active in several organs and tissues of adult female and male mice. The first 0.4 kilobases of 5'-flanking DNA (up to -364) are sufficient to direct widespread expression of the transgene in mouse organs. This indicates that genetic elements functional in various cell types are included in this segment. Furthermore, the first exon and intron of the mER gene are necessary to achieve sexually dimorphic expression of the transgene in neurons located at specific sites within the central nervous system. These mER promoter transgenic mice will be useful in mapping estrogen- responsive cell types under different physiological and pathological conditions in vivo, in defining ontogenesis of estrogen action in the mouse, and in studying the mechanisms that regulate ER gene transcription.

Antioxidant and protective effect of latex of Calotropis procera against alloxan-induced diabetes in rats.

In the present study, dry latex (DL) of Calotropis procera possessing potent anti-inflammatory activity was evaluated for its antioxidant and anti-hyperglycemic effects against alloxan-induced diabetes in rats. Daily oral administration of DL at 100 and 400 mg/kg doses produced a dose-dependent decrease in the blood glucose and increase in the hepatic glycogen content. DL also prevented the loss of body weight in diabetic rats and brought down the daily water consumption to values comparable to normal rats. DL also produced an increase in the hepatic levels of the endogenous antioxidants, namely superoxide dismutase (SOD), catalase and glutathione, while it brought down the levels of thiobarbituric acid-reactive substances (TBARS) in alloxan-induced diabetic rats. The efficacy of DL as an antioxidant and as an anti-diabetic agent was comparable to the standard anti-diabetic drug, glibenclamide.

Protective effect of proteins derived from Calotropis procera latex against acute inflammation in rat.

The non-dialysable proteins present in the latex of plant Calotropis procera possess anti-inflammatory and analgesic properties. The aim of this study was to evaluate the effect of latex proteins (LP) on the level of inflammatory mediators, oxidative stress markers and tissue histology in the rat model of carrageenan-induced acute inflammation. This study also aimed at evaluating the anti-inflammatory efficacy of LP against different mediators and comparing it with their respective antagonists. Paw inflammation was induced by subplantar injection of carrageenan, and the effect of LP was evaluated on oedema volume, level of TNF-α, PGE(2), myeloperoxidase, nitric oxide, reduced glutathione, thiobarbituric acid-reactive substances and tissue histology at the time of peak inflammation. Paw inflammation was also induced by histamine, serotonin, bradykinin and PGE(2), and the inhibitory effect of LP against these mediators was compared with their respective antagonists at the time of peak effect. Treatment with LP produced a dose-dependent inhibition of oedema formation, and its anti-inflammatory effect against carrageenan-induced paw inflammation was accompanied by reduction in the levels of inflammatory mediators, oxidative stress markers and normalization of tissue architecture. LP also produced a dose-dependent inhibition of oedema formation induced by different inflammatory mediators, and its efficacy was comparable to their respective antagonists and more pronounced than that of diclofenac. Thus, our study shows that LP has a potential to be used for the treatment of various inflammatory conditions where the role of these mediators is well established.

Comparative analysis of epidermal growth factor receptor mRNA levels in normal, benign hyperplastic and carcinomatous prostate.

Epidermal growth factor receptor (EGFR), a mediator of mitogenic activity of epidermal growth factor and transforming growth factor-alpha, has been shown to be associated with tumour progression. We have detected a level of EGFR transcript in normal, hyperplastic (BPH) and carcinomatous (CaP) prostate tissues by cytoplasmic dot hybridization. Our results show that the majority of CaP cases (62% of untreated cases and 71% of treated cases) were positive for EGFR mRNA while about 50% of normal and BPH cases were positive for EGFR mRNA. The level of EGFR transcript was significantly higher in the untreated CaP group as compared to the normal group (P < 0.05), while the difference in the normal and BPH groups was not statistically significant.

Inflammation induced by latex of Calotropis procera--a new model to evaluate anti-inflammatory drugs.

Latex of Calotropis procera was studied for its inflammatory reactions using pedal oedema and air pouch models of inflammation in rats. Subcutaneous injection of aqueous solution (0.1 ml of 1%) of dry latex (DL) into the plantar surface of paw produced significant inflammation. Maximum inflammatory response was obtained 1 h after the injection and was maintained for a further 1 h. The inflammatory response was accompanied by an increase in vascular permeability that reached its maximum within 15 min. Inflammation was also induced in the 6-day-old rat air pouch by injecting a 2.5% solution of DL. The latter model was characterized for the exudate volume and its protein concentration, and wet and dry weights of granuloma. A time-course study indicated that both the exudate volume and the weight of granuloma were at maximum on day 5 after DL injection while the protein concentration peaked on the third day. Further, the two models were also studied for the anti-inflammatory effect of various drugs. It was observed that in the pedal oedema model, phenylbutazone was more effective than prednisolone while almost complete inhibition was produced by mepyramine and cyproheptadine. On the other hand, in the air pouch model, prednisolone was more effective than phenylbutazone in inhibiting the inflammation. Thus, the DL-induced inflammation in different models could be used to evaluate anti-inflammatory drugs.

In vivo modulation of androgen receptor by androgens.

AIM: To study the effect of androgen and antiandrogen on the level of androgen receptor (AR) mRNA. METHODS: The total RNA was extracted from the prostate and analyzed by slot blot analysis. The blots were hybridized with AR cDNA probe and 1A probe (internal control) and autoradiography was performed. The intensity of signal was measured with a densitometer and the ratio of AR RNA and 1A RNA was calculated. RESULTS: Androgenic deprivation produced by castration decreased the weight of the prostate and increased the levels of AR mRNA. Treatment of the castrated rats with testostrone increased the weight of prostate and decreased the levels of AR mRNA. Treatment of normal rats with flutamide decreased the weight of the gland and increased the levels of AR mRNA. CONCLUSION: Androgens produce proliferative effect on the prostate and negatively regulate the AR transcription.

Anti-inflammatory activity of the latex of Calotropis procera.

The anti-inflammatory property of the latex of Calotropis procera was studied on carrageenin- and formalin-induced rat paw oedema model. A single dose of the aqueous suspension of the dried latex was effective to a significant level against the acute inflammatory response.

In vivo and in vitro effect of latex of Calotropis procera on gastrointestinal smooth muscles.

The present study was carried out to evaluate the effect of dry latex (DL) of Calotropis procera on smooth muscles of gastrointestinal tract. Oral administration of DL to rats (50-1000 mg/kg) produced a dose-dependent decrease in intestinal transit along with a decrease in intestinal content as compared to control group. At lower doses DL produced dose-dependent contractions of gastrointestinal smooth muscles in vitro (rabbit ileum and fundus of rat stomach) that was followed by desensitization at higher doses.

Preliminary studies on the analgesic activity of latex of Calotropris procera.

In this study we have evaluated the analgesic activity of dry latex (DL) of Calotropis procera. A single oral dose of DL ranging from 165 to 830 mg/kg produced a significant dose dependent analgesic effect against acetic acid induced writhings. The effect of DL at a dose of 415 mg/kg was more pronounced as compared to a 100 mg/kg oral dose of aspirin. On the other hand DL (830 mg/kg) produced marginal analgesia in a tail-flick model which was comparable to aspirin. The analgesic effect of DL was delayed by 1 h by naloxone at a dose of 0. 5 mg/kg, i.p., which completely blocked the analgesic effect of morphine (10 mg/kg, i.p.). However, the effect of aspirin was not blocked by naloxone. The 830 mg/kg oral dose of DL did not produce toxic effects in mice and the LD(50) was found to be 3 g/kg.

Anti-diarrhoeal activity of the latex of Calotropis procera.

The dry latex (DL) of Calotropis procera (Asclepiadaceae), a potent anti-inflammatory agent has been evaluated for anti-diarrhoeal activity. Like atropine and phenylbutazone (PBZ), a single oral dose of DL (500 mg/kg) produced a significant decrease in frequency of defecation, severity of diarrhoea and afforded protection from diarrhoea in 80% rats treated with castor oil. To understand the mechanism of its anti-diarrhoeal activity, we further evaluated its effect on intestinal transit, castor oil induced intestinal fluid accumulation (enteropooling) and electrolyte concentration in the intestinal fluid. DL produced a decrease in intestinal transit (27-37%) as compared to both normal and castor oil treated animals. Unlike atropine, DL significantly inhibited castor oil induced enteropooling. However, it did not alter the electrolyte concentration in the intestinal fluid as compared to castor oil treated rats.

Gene therapy.

Gene therapy replaces or supplements the activity of a resident defective or missing gene by a cloned functional one. A functional gene may be delivered to the target tissues ex vivo or in vivo using various physical methods and several chemical and biological vehicles. In this article we present an overview of different experimental models and the progress made in clinical studies on gene therapy and discuss the prospects of the antisense approach and the ethical concerns related to the technique.

Prostate gland: structure, functions and regulation.

The prostate gland plays an important role in male reproduction. It secretes enzymes, lipids, amines and metal ions essential for the normal function of spermatozoa. Development, differentiation and maintenance of the prostate gland depend on steroid and peptide hormones. Beside hormones growth factors also regulate the prostate gland. This review will focus on the structure, functions and mode of regulation of the prostate gland.