Sperm abnormalities induced by pre-pubertal exposure to cyclophosphamide are effectively mitigated by Moringa oleifera leaf extract.

Moringa oleifera L. is a medicinal plant with potential antioxidant property. This study was aimed at investigating the chemoprotective effect of Moringa oleifera leaf extract (MOE) on cyclophosphamide (CP)-induced testicular toxicity. Two-week-old male Swiss albino mice were intraperitoneally injected with phosphate-buffered saline, 50 mg kg(-1) of CP and 25 mg kg(-1) of MOE. In combination treatment, mice were injected with 25 mg kg(-1) of MOE 24 h prior to CP injection, 24 h prior and post-CP injection and 24 h post-CP injection for 5 consecutive days (10 mg kg(-1) ). Six weeks later, mice were sacrificed to assess epididymal sperm parameters. MOE alone did not have any significant effect on sperm parameters. However, acute injection of CP resulted in significant decline in motility (P < 0.001), increase in head abnormality (P < 0.01) and DNA damage (P < 0.05). Combining MOE with CP increased the sperm density, motility and reduced head defect and DNA damage, irrespective of the schedule and dosage of MOE. Administration of MOE prior to CP significantly elevated the level of superoxide dismutase and catalase with concomitant decrease in lipid peroxidation in the testicular tissue. In conclusion, MOE may have potential benefit in reducing the loss of male gonadal function following chemotherapy.

Iontophoresis-mediated transdermal permeation of peptide dendrimers across human epidermis.

PURPOSE OF THE STUDY: The overall aim of the present work was to elucidate the effects of iontophoresis on assisting permeation/deposition of peptide dendrimers across/within human skin. PROCEDURES: A series of peptide dendrimers containing arginine and histidine as terminal acids were synthesized and characterized. These dendrimers were subjected to passive and iontophoretic permeation studies across human epidermis. RESULTS: The synthesized peptide dendrimers were found to be stable in epidermal, dermal and skin extracts up to 6 h. Passive diffusion studies revealed that none of the synthesized peptide dendrimers permeated human epidermis up to 6 h, although minute concentrations of low molecular weight dendrimers were detected in receptor medium at the end of 24 h. Application of iontophoresis significantly increased the permeation of all the tested peptide dendrimers across human skin in a molecular weight-dependent manner compared to simple passive diffusion. Electromigration was found to be the dominant mechanism behind the iontophoretic permeation of peptide dendrimers across human skin. CONCLUSIONS: The present study demonstrates that iontophoresis is an effective technique in enhancing the transdermal permeation of peptide dendrimers. MESSAGE OF THE PAPER: This study foresees the possibility of applying peptide dendrimers in iontophoretic delivery of drugs and macromolecules across/within the skin.

Development and validation of a reversed-phase high-performance liquid chromatographic method for quantification of peptide dendrimers in human skin permeation experiments.

The aim of the present work was to develop and validate a simple RP-HPLC method with UV detection to quantify peptide dendrimers in skin permeation experiments. Six dendrimers of varying positive charges (4(+), 8(+) and 16(+)) containing either histidine or arginine as terminal aminoacids were prepared by solid phase peptide synthesis. Mobile phase containing 0.02% (v/v) heptafluorobutyric acid in 90% acetonitrile-water was capable of separating all dendrimers from interfering peaks of receptor fluid. For the calibration of each dendrimer, a different dendrimer from the same class was selected as the internal standard. The results of preliminary human skin permeation studies showed that the developed analytical method can be successfully used for the quantification of cationic poly(aminoacid)-based dendrimers in skin permeation experiments.

Preparation and physicochemical and preclinical evaluations of recrystallized celecoxib.

The aim of the present study is to increase the aqueous solubility of celecoxib by recrystallization from distilled water, Tween-80, and polyethylene glycol-400. The prepared crystals were evaluated for various physicochemical evaluations, dissolution rate, and in vivo performance like analgesic activity (by writhing and hot plate method) and pharmacokinetics in mice. The practical yield of the crystals ranged between 83 and 98%, and celecoxib content was more than 99%. Celecoxib showed an almost 5-fold increase in solubility when recrystallized in the presence of Tween-80 (2%). The dissolution rates of celecoxib from the co-crystal forms were considerably higher than that of plain celecoxib. The infrared and differential scanning calorimetry studies indicated the absence of a well-defined interaction between celecoxib and carriers. The differential scanning calorimetry and X-ray diffraction studies indicated the amorphization or partial amorphization of the drug. The scanning electron microscopy showed fluffy, porous, and fine particles in recrystallized celecoxib. The particle size of prepared co-crystals was considerably reduced in comparison with plain celecoxib. The crystals prepared with Tween-80 (2%) showed significantly higher analgesic activity than plain celecoxib. In pharmacokinetic study, the prepared crystals exhibited significantly high and rapid absorption along with improved bioavailability.

Preparation, in vitro, preclinical and clinical evaluations of once daily sustained release tablets of aceclofenac.

The objective of the present study was to develop "once daily" sustained release tablets of aceclofenac by direct compression using hydroxypropyl methylcellulose-K4M (HPMC). The solubility studies of aceclofenac were conducted to select suitable dissolution media. The drug-excipient mixtures were subjected to preformulation studies. The tablets were subjected to physicochemical, in vitro drug release and stability studies. Preclinical (anti-inflammatory, analgesic, pharmacokinetic and toxicity studies) and clinical pharmacokinetic studies were conducted for optimized tablets. Based on the preformulation results, microcrystalline cellulose (MCC), dicalcium phosphate and spray dried lactose (SDL) were selected as directly compressible vehicles. Because of the incompatibility with aceclofenac, SDL was excluded from the study. The physicochemical properties of tablets were found within the limits. By comparing the dissolution profiles with the marketed product, the tablet containing HPMC (45%) and MCC (30%) along with talc and magnesium stearate (1% w/w, each) (Tablet B7) was considered as a better formulation. This tablet exhibited almost similar drug release profile in different dissolution media as that of marketed tablet. Tablet B7 was stable in accelerated conditions for 6 months. The composition of this tablet showed almost similar preclinical pharmacological activities compared to marketed tablet composition and did not exhibit any toxicity in rats and mice with respect to tested haematological and biochemical parameters along with body weight, food and water intake. The pharmacokinetic study in healthy human volunteers indicated that B7 tablet produced an extended drug release of drug upto 24 h as that of marketed product with almost identical pharmacokinetic parameters.

In Vitro and In Vivo preparation evaluations of bleomycin implants and microspheres Prepared with DL-poly (lactide-co-glycolide).

In this investigation, poly(lactide-co-glycolide) (PLGA) gel implants and microspheric depot systems of bleomycin (BLM) were formulated and evaluated in vivo in mice bearing transplantable solid tumor (fibrosarcoma). The pharmacodynamic studies showed that both the formulations retarded tumor growth significantly (p<0.05) when compared to the control animals (without any drug treatment). Preliminary pharmacokinetic studies illustrated controlled release of the drug into the systemic circulation to elicit the anti-neoplastic action. The gel implants showed better release characteristics and greater pharmacodynamic action when compared to the microspheres, thus demonstrating the feasibility of employing biodegradable depot polymer gel matrix for chronic cancer therapy.

Effect of Dianex, a herbal formulation on experimentally induced diabetes mellitus.

Dianex, a polyherbal formulation consisting of the aqueous extracts of Gymnema sylvestre, Eugenia jambolana, Momordica charantia Azadirachta indica, Cassia auriculata, Aegle marmelose, Withania somnifera and Curcuma longa was screened for hypoglycemic activity in normal and streptozotocin induced diabetic mice. Dianex was administered in different doses of 100-500 mg/kg/day orally in acute (6 h) and long-term (6 weeks) studies. Blood glucose levels were checked 2-6 h after treatment in acute studies and every 2 weeks in long-term studies. Body weight was recorded on the first and final day of the treatment in the long-term studies with diabetic mice. After 6 weeks, high-density lipoprotein, triglycerides, total cholesterol, alanine transaminase (ALT), aspertate transaminase (AST), urea and creatinine were estimated in serum of the diabetic mice. Glycogen and total protein levels were estimated in the liver. Also, the liver and pancreas was subjected to histological examination. Oral glucose tolerance and in vitro free radical scavenging activity was also studied. Dianex produced significant (p<0.05) hypoglycemic activity at 250-500 mg/kg doses in both normal and diabetic mice in acute and long-term studies. The body weight of diabetic mice significantly (p<0.05) increased with all tested doses of Dianex. The elevated triglycerides, cholesterol, ALT, AST, urea and creatinine levels in diabetic mice were significantly (p<0.05) reduced at the doses of 250 and 500 mg/kg. The liver glycogen and protein levels were both significantly (p<0.05) increased in diabetic mice at 250 and 500 mg/kg doses. Dianex increased the glucose tolerance significantly (p<0.05) in both normal and diabetic mice at all the doses tested. Histopathological examination showed that the formulation decreased streptozotocin induced injury to the tissues at all the doses tested. It produced significant (p<0.05) free radical scavenging activity against ABTS+, DPPH and hydroxyl free radicals at the concentrations ranging between 10-1000 microg/ml.Thus, in the present study, Dianex produced significant hypoglycemic activity in both normal and diabetic animals. It also reversed other diabetic complications in diabetic mice at 250 and 500 mg/kg doses. In our earlier study, Dianex was well tolerated in laboratory animals at higher doses (upto 10 g/kg in mice, acute toxicity; up to 2.5 g/kg in rats, subacute toxicity studies for 30 days) without exhibiting any toxic manifestation. Hence, Dianex may be useful in the treatment of diabetes mellitus.

Glibenclamide transdermal patches: physicochemical, pharmacodynamic, and pharmacokinetic evaluations.

In the present study, matrix type transdermal patches containing glibenclamide were prepared using different ratios of ethyl cellulose (EC)/polyvinylpyrrolidone (PVP) and Eudragit RL-100 (ERL)/Eudragit RS-100 (ERS) by solvent evaporation technique. The possible drug and polymer interaction was studied by infrared spectroscopy, differential scanning calorimetry, and HPTLC analysis. All the prepared formulations were subjected to physicochemical studies (thickness, weight variation, drug content, moisture content and uptake, and flatness), in vitro release and in vitro permeation studies through mouse skin. The results suggested that there was no interaction between drug and polymers. Variations in drug release/permeation profiles among the formulations studied were observed. The microphotographs obtained by scanning electron microscopy showed the formation of pores on the surface of the patches after in vitro skin permeation studies. Based on physicochemical and in vitro skin permeation studies, the formulations with EC:PVP (3:2) and ERL:ERS (4:1) were selected for in vivo experiments. The hypoglycemic activity of the patches in comparison with oral glibenclamide administration was studied for acute (24 h) and long-term (6 weeks) effect in both normal and streptozotocin-induced diabetic mice. Various biochemical parameters (serum levels of high-density lipoprotein-cholesterol, triglycerides, total cholesterol, alanine transaminase, aspertate transaminase, urea, and creatinine and liver protein and glycogen content) and histopathological (liver, pancreas and stomach) studies were carried out in diabetic mice after treating for 6 weeks. The patches were subjected to skin irritation test (by both visual observation and histopathological evaluation), oral glucose tolerance test and pharmacokinetic evaluation in mice. The results revealed that the patches successfully prevented the severe hypoglycemia in the initial hours, which is the major side effect associated with oral route. The patches maintained similar effect during long-term treatment also. The transdermal systems produced better improvement with all the tested biochemical parameters compared to oral administration. They produced improved repair of the tissues after diabetes induced tissue injury and exhibited negligible skin irritation. The pharmacokinetic evaluation showed that the patches could maintain almost steady-state concentration of drug within the pharmacologically effective range for prolonged period of time. The better in vivo performance of the transdermal patches of glibenclamide in comparison with oral administration could be due to day-to-day glycemic control on long-term application.

A toxicological evaluation of 2-dimethylaminomethyl-1-phenyl-2-propen-1-one hydrochloride.

2-Dimethylaminomethyl-1-phenyl-2-propen-1-one hydrochloride (3) is a novel cytotoxic and anticancer agent. The objective of this study was to obtain information pertaining to possible toxic symptoms detected by in vivo evaluations in mice and an in vitro test for mutagenicity. The data obtained revealed that 3 had no effect on alanine transaminase, aspartate transaminase, HDL cholesterol and protein concentrations in sera nor were variations in the numbers of red and white blood cells detected. Furthermore autopsies of treated mice revealed no pathological symptoms in the heart, kidney, brain, spleen and testes. However elevation of the concentrations of total cholesterol, triglycerides, creatinine and urea were noted in treated mice as well as inflammation of the liver and lungs. Chromosomal aberrations were detected in a micronuclei test. In the Ames test, compound 3 was converted into one or more mutagens in the presence (but not the absence) of a murine liver homogenate. Thus future molecular modifications of 3 should bear in mind approaches to reduce or minimize unwanted side effects.

Effect of some penetration enhancers on the permeation of glibenclamide and glipizide through mouse skin.

The purpose of this investigation was to study the effect of some penetration enhancers on in vitro permeation of glibenclamide and glipizide through mouse skin. Ethanol in various concentrations, N-methyl-2-pyrrolidinone, transcutol, propylene glycol and terpenes like citral, geraniol and eugenol were used as penetration enhancers. The in vitro skin permeation experiments were conducted by both simultaneous application of drug and enhancer solution and by pretreatment of the skin with neat enhancer. At the end of the experiment drug retained in the skin was estimated. The flux values (microg/cm2/h) of both drugs significantly (p < 0.05) increased in the presence of penetration enhancers, except transcutol and propylene glycol. The glibenclamide flux values ranged from 1.42 +/- 0.09 without enhancer, to 18.25 +/- 1.21 in a combination of 50% ethanol and 5% eugenol. Glipizide flux values ranged from 3.21 +/- 0.51 without enhancer, to 57.21 +/- 5.25 in a combination of 50% ethanol and 5% eugenol. Skin retention and solubility of both drugs increased with all penetration enhancers compared to control (except propylene glycol). As the target permeation rates for glibenclamide and glipizide were calculated to be 193.8 and 184.8 microg/h respectively, the present study showed that the required permeation rates for both drugs could be achieved with the aid of enhancers by increasing the area of application in an appreciable range.

Preliminary studies on acute and subacute toxicity of an antidiabetic herbal preparation, Dianex.

Dianex, a polyherbal formulation intended to use for diabetic patients, has been screened for toxic effects. For acute toxicity studies, Dianex was administered orally in graded doses of 0.75-10 g/kg to the mice. For subacute toxicity studies, different doses of Dianex (1.0, 1.5 and 2.5 g/kg) were administered orally to the rats once daily for 30 days. Animals were observed for physiological and behavioural responses, mortality, food and water intake and body weight changes. Hematological evaluation was performed weekly. All the animals were sacrificed on 31st day and changes in organ weights and histology were examined. Biochemical studies were done in liver and serum. No mortality was observed up to 10 g/kg of Dianex in acute toxicity study. Daily administration of as high as 2.5 g/kg dose of Dianex did not result in any mortality or changes in gross behaviour, body weight, weight and histology of different organs or serum and liver biochemistry. However, significant increase in RBC count and hemoglobin level was observed in the treated animals at all doses. Other peripheral blood constituents were in the normal range. The dose of Dianex to produce significant antidiabetic activity in mouse, 0.25-0.5 g/kg, is much lower than the doses used in the present study. Therefore such doses may be safe for daily administration without causing any serious side effects.

Transdermal delivery of glibenclamide and glipizide: in vitro permeation studies through mouse skin.

The purpose of this investigation was to study the feasibility of transdermal delivery of glibenclamide and glipizide. In vitro permeation of these drugs was studied through mouse skin using various penetration enhancers like Tween -20, polyethyleneglycol-400, ethanol and d-limonene by simultaneous application of drug and enhancer solution or by pretreatment of the skin with neat enhancer. The partition coefficient values indicated that both drugs partition well into the skin. Glipizide did not show any skin metabolism, while glibenclamide showed a minimal metabolism during in vitro skin metabolism studies. The flux values (microgram/cm2/h) of both drugs significantly (p < 0.05) increased in the presence of penetration enhancers. The glibenclamide flux values ranged from 1.39 +/- 0.13 without enhancer, to 19.01 +/- 2.14 in a combination of 50% ethanol and 5% d-limonene. Glipizide flux values ranged from 3.01 +/- 0.74 without enhancer, to 62.97 +/- 7.10 in a combination of 50% ethanol and 5% d-limonene. Skin retention and solubility of both drugs increased with all penetration enhancers compared to control. The target permeation rates for glibenclamide and glipizide were calculated to be 193.8 and 184.8 micrograms/h respectively. The present study showed that the target permeation rates for both drugs could be achieved with the aid of enhancers by increasing the area of application in an appreciable range.

Angina bullosa hemorrhagica (recurrent oral haemophlyctenosis).

A case of recurrent oral haemophlyctenosis, characterized by sudden onset of tense blisters on the tongue is being reported. The blisters were fillcd with haemorrhagic fluid and healed within a week.

Surgical management of stable vitiligo: A review with personal experience.

BACKGROUND: Various modalities are available for surgical management of stable, localized patches of vitiligo, which are resistant to conventional medical treatment. Cutaneous surgeons often struggle to select among various methods of surgical treatment that include camouflage tattooing, melanocyte transplants, excision, and melanocyte culture. The advantages, disadvantages, and limitations of all the available modalities are reviewed. OBJECTIVE: The purpose of this review is to find out if any guidelines can be drawn regarding the surgical management of stable vitiligo. METHODS: This article is based on a review of the medical literature and the author's personal experience over the last decade. RESULTS: It is difficult to draw any fixed guidelines from this review. Treatment must be individualized depending upon the site and the extent of involvement. CONCLUSION: Although the retrospective nature of this article limits its validity, discussing the various therapeutic options is of value as continuing medical education. Surgical treatment may be individualized to obtain the best possible cosmetic result.

Another method of tie-over dressing for surgical wounds of hair-bearing areas.

BACKGROUND: We describe a modified tie-over dressing for any kind of surgical wound in the hair-bearing areas. METHODS: The long ends of the sutures are tied over the gauze pad to secure it. CONCLUSIONS: This modified technique of tie-over dressing can be utilized for the majority of surgical wounds in hair-bearing areas, such as the scalp. The dressing can be removed 1 or 2 days postoperatively by the patients, and the wound can be washed thereafter. The dressing helps ensure hemostasis, is simple to perform, looks tidy, and is well accepted by the patients.

Manual punch for tattooing.

We have devised and designed a manual instrument for tattooing with two shapes. It has been found to have certain advantages over the routinely used electrically operated machine. This instrument can be sterilized easily.