Advancements in Nanotechnology for Enhanced Antifungal Drug Delivery: A Comprehensive Review.

Infections caused by fungi can be mildly bothersome or fatal, causing life-threatening conditions or even death. Antifungal drugs have used synthetic chemicals, organic compounds, and phytoconstituents in their formulations to treat fungal infections. Research into novel antifungal drugs has progressed more rapidly than into antibacterial treatments. This can be attributed to the low resistance of fungal infections to antifungal bioactivities and the relatively low incidence of these diseases. Carrier systems based on nanotechnology have generated much interest recently because of the incredible potential of these systems. By using nanoarchitecture as a better carrier and drug delivery system (DDS), we can have greater antifungal effectiveness, bioavailability, targeted action, and less cytotoxicity, a development made possible using nanotechnology. This review discusses various nanocarrier-based technologies in addition to other nanotechnological methods. These include liposomes, transfersomes, ethosomes, niosomes, dendrimers, polymeric nanoparticles, polymer nanocomposites, metallic nanoparticles, carbon nanomaterials, etc. This review focused on general information regarding fungi infections, different antifungal agent types and mechanisms of action, and an overview of formulation strategies such as nanotechnology systems, which are frequently researched for antifungal therapies. We concluded that new drug delivery systems are crucial to delivering antifungal medicines to their target site with the optimum concentration. The researchers also concentrated on these innovative drug delivery systems, which primarily focus on regulating and maintaining the release of antifungal drugs.

Establishment of antitick efficacy of a phytoformulation prepared from Annona squamosa leaf extracts for the management of acaricide resistant tick infestations on cattle.

To combat the problem of acaricide resistance in ticks, efforts have been made to develop eco-friendly herbal acaricides which are safe for animal use with no possibilities of evoking environment and residual toxicity. In the present study, the acaricidal properties of a commonly available plant, Annona squamosa have been established to develop a novel antitick phytoformulation using leaf extract, suitable solvents and emulsifier for managing resistant tick infestations in animals. The quality control of the safe and stable phytoformulation using Rutin as a major constituent was maintained by HPTLC profiling. Several phytformulations (A-E) were prepared and formulation 'C' is identifried as the most effective showing 52.5-75% antitick activity against in vitro treated resistant strains of R. microplus (IVRI-IV and V) with 33.8-40.2% inhibition of oviposition while 65-85% mortality against multiacaricide resistant field populations. The phytoformulation provided 70.4% efficacy after first larval challenge in experimentally infested animals. The field efficacy of the formulation in different multilocational field trials was 68.53-77.8%. The detail antitick efficacy of the formulation along with safety and stability are explained as a component of antitck technology to manage resistant tick problem in livestock.

In Vitro and In Vivo Antifilarial Activity of Standardized Extract of Calotropis procera Flowers against Brugia malayi.

BACKGROUND: Lymphatic filariasis (LF) is a parasitic disease that causes permanent disability (elephantiasis). Currently used antifilarial drugs are failing to control LF and there is resurgence in some areas. Looking for new antifilarial leads, we found that Calotropis procera plant parts have been used in traditional medicine for alleviating elephantiasis but the antifilarial activity is not known. OBJECTIVE: In the present study, the antifilarial activity of ethanolic extract (A001) and its hexane fraction (F001) of C. procera flowers was investigated using the human filarial parasite Brugia malayi. METHODS: A001 and F001 were tested for antifilarial activity using motility and 3-(4,5-dimethylthiazol-2- yl)-2,5 diphenyltetrazolium bromide (MTT) assays (in vitro) and in the rodent models B. malayi- Meriones unguiculatus and B. malayi-Mastomys coucha. In the rodent models, A001 and F001 were administered orally for 5 consecutive days, and the adult worm burden and course of microfilaraemia were determined. RESULTS: Both A001 and F001 showed microfilaricidal and macrofilaricidal activity in vitro. In animal models, A001 killed ~49-54% adult worms. In M. coucha model, F001 killed 12-60% adult worms in a dose (125-500 mg/kg) dependent manner; A001 and F001 suppressed microfilaraemia till days 91 and 35 post initiation of treatment, respectively. HPTLC revealed 0.61% lupeol, 0.50% ß-sitosterol and 1.50% triacontanol in F001. CONCLUSION: Flowers of C. procera have definite microfilaricidal and macrofilaricidal activities. Whether this activity is due to lupeol, ß-sitosterol and triacontanol found in the hexane fraction remains to be investigated. This is the first report on the antifilarial efficacy of flowers of the plant C. procera.

A Validated HPTLC Densitometric Method for Determination of Lupeol, ß-Sitosterol and Rotenone in Tephrosia purpurea: A Seasonal Study.

Tephrosia purpurea (L.) Pers., commonly known as "sarpunkha" and "wild indigo", is being used in traditional systems of medicine to treat liver disorders, spleen and kidney. In the present study, a validated High Performance Thin Layer Chromatography (HPTLC) method was established for the estimation of lupeol, ß-sitosterol and rotenone in various extracts of T. purpurea with the aim to see the effect of seasons on the quantity of aforesaid phytoconstituents. The plant material was collected in summer (April), rainy (August) and winter (December) during 2013-2014 from Lucknow, India. The method was validated in terms of precision, repeatability, specificity, sensitivity linearity and robustness. The method permits reliable quantification and showed good resolution on silica gel with toluene-ethyl acetate-formic acid (9:1:1 v/v/v) as mobile phase, and characteristic bands of ß-sitosterol, rotenone and lupeol were observed at Rf 0.38, 0.45 and 0.52, respectively. The content of aforesaid phytoconstituents varies from season to season and extract to extract. Our finding indicated that winter season (December) may not be appropriate for collection of T. purpurea for the preparation of therapeutic formulations because of the high content of rotenone, a known insecticide that is responsible for Parkinson's disease and associated with heart failure, fatty liver and liver necrosis.

Ethnopharmacological uses, phytochemistry and pharmacology of genus Adiantum: A comprehensive review.

ETHNOPHARMACOLOGICAL RELEVANCE: Genus Adiantum (Pteridaceae) forms a significant dominant component of many plant communities especially in the tropical and temperate regions. These are commonly known as maidenhair ferns and several have been used medicinally in different parts of the world. They exhibit antidysenteric, antiulcer, antimicrobial, antitumor and antiviral activities. The traditional uses of Adiantum species are known to be for respiratory problems such as cough cold, fever, pneumonia and mucous formation. AIM OF THE REVIEW: This review aims to provide a comprehensive and updated, categorized information on the botanical aspects, traditional uses, phytochemistry, pharmacological activities and toxicological research of Adiantum species in order to explore their therapeutic potential and evaluate future research opportunities. MATERIALS AND METHODS: The available information on various species belonging to the genus Adiantum was collected via electronic search (using Pubmed, SciFinder, Scirus, Google Scholar, JCCC@INSTIRC and Web of Science) and a library search for articles published in peer-reviewed journals. RESULTS: The literature provided information on several ethnopharmacologically well known Adiantum species, the best studied species being A. capillus-veneris. From these plant species, more than 130 compounds belonging to triterpenoids, flavanoids, phenyl propanoids, phenolics, coumarins, phytosterols, fatty acids and others were identified. Experimental evidences confirmed that the Adiantum species could be used in treating microbial infections, diabetes, liver disorders as well as inflammatory disorders. CONCLUSIONS: Plants belonging to the genus Adiantum have offered bioactive crude extracts as well as pure compounds, thus substantiating their effectiveness in traditional medicine. Although toxicity studies carried out on some Adiantum species have showed them to be non-toxic, further toxicological studies are still required to confirm their safety in humans. Future research should be directed towards implementing an integrated approach through intensive investigations of all the species of Adiantum relating to phytochemical and pharmacological properties, especially bio-assay guided isolation of phytoconstituents, their mechanism of action, as well as their bioavailability and pharmacokinetics, safety and efficacy.

Oleanolic-bioenhancer coloaded chitosan modified nanocarriers attenuate breast cancer cells by multimode mechanism and preserve female fertility.

Addressing multidrug resistant stage of breast cancer is an impediment for chemotherapy. Moreover, breast cancer chemotherapy has potential enduring confrontations i.e. related toxicity including effect on fertility of young female patients. The co-delivery of polyphenolic bio-enhancers with oleanolic acid in chitosan coated PLGA nanoparticles was designed for oral delivery with enhanced antitumor effect consecutively preserving the female fertility. The optimized oleanolic- bio-enhancer nano formulation CH-OA-B-PLGA with particle size was 342.2±3.7nm and zeta potential of 34.2±3.1mV was capable of lowering viability in MDAMB 231 cell line 16 times than OA. Further, mechanistic studies in MDAMB-231 cells revealed that CH-OA-PLGA induces apoptosis by mitochondrial membrane disruption; follows ROS mediated and caspase dependent apoptosis. The antitumor effect studied in 4-T1 induced Balb/c mice mammary tumor model displayed augmented antitumor potency by CH-OA-B-PLGA in comparison to OA. In the in vivo toxicity on Sprague-Dawley rat model, CH-OA-B-PLGA significantly displayed the safe profile and also preserves fertility in female rats. The experiment result suggests co-delivery of oleanolic acid with bio-enhancers as a breakthrough for developing safe chemotherapy for hormone independent breast cancer therapy countering the toxicity issues.

Antioxidant and Hepatoprotective Potential of Phenol-Rich Fraction of Juniperus communis Linn. Leaves.

BACKGROUND: Juniperus communis Linn. is an important plant in India traditional system of medicine which is widely used by different tribes in many countries. OBJECTIVE: In the present study, the antioxidant, cytotoxic and hepatoprotective activities of Juniperus communis leaves were investigated against various models. MATERIALS AND METHODS: ethanolic extract (70% v/v) of J. communis leaves was successively extracted using hexane and ethyl acetate to prepare various fractions. Total phenol content was resolute by the Folin-Ciocalteau's process. The antioxidant properties of the different fractions/extract of leaves of J. communis were examined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and Fe2+ chelating ability. Cytotoxic activity was examined by cell viability assay on HepG2 cells. Hepatoprotective activity of ethyl acetate fraction (EAF) evaluated against PCM-Paracetamol-induced hepatic damage in Wistar albino rats. RESULTS: Total phenol content was found maximum 315.33 mg/GAE/g in EAF. Significant scavenging activity were found for EAF (IC50 = 177 µg/ml) as compared to standard BHT (IC50 = 138 µg/ml), while EAF showed good Fe2+ chelating ability having an IC50 value of 261 mg/ML compared to standard ethylenediaminetetraacetic acid (7.7 mg/mL). It was found that EAF treated group shows remarkable decrease in serum Aspartate aminotransferase, serum Alanine aminotransferase, total bilirubin, direct bilirubin, and alkaline phosphatase level in treatment group as compared to the hepatotoxic group. CONCLUSION: EAF of J. communis leaves is found to be potent antioxidant and hepatoprotective without any cytotoxicity and it can also be included in nutraceuticals with notable benefits for mankind or animal health. SUMMARY: Phenol-rich fraction (PRF) and other fractions/extract of Juniperus communis leaves were screened for antioxidant, cytotoxic, and hepatoprotective activity.Significant antioxidant and hepatoprotective activity without any cytotoxicity were found while treating with ethyl acetate fraction (EAF). Abbreviations used: HepG2: Liver hepatocellular carcinoma, BHT: Butylated hydroxytoluene, PCM: Paracetamol, IC50: Half maximal inhibitory concentration, RSA: Radical Scavenging Activity, WST: Water-soluble tetrazolium.

Spices: Therapeutic Potential in Cardiovascular Health.

BACKGROUND: Dietary factors play a key role in the development as well as prevention of certain human diseases, including cardiovascular diseases. Currently there has been an increase in global interest to identify medicinal plants that are pharmacologically effective and have low or no side effects for use in preventive medicine. Culinary herbs and spices are an important part of human nutrition in all the cultures of the world. There is a growing amount of literature concerning the potential benefits of these herbs and spices from a health perspective especially in conferring protection against cardiovascular diseases. OBJECTIVE: The objective of this review is to provide information on the recent scientific findings on some common spices that have a distinct place in folk medicine in several of the Asian countries as well as on their traditional uses for the role they can play in the management of heart diseases and which may be useful in defining cost effective and inexpensive interventions for the prevention and control of CVDs. METHOD: Systematic literature searches were carried out and the available information on various medicinal plants traditionally used for cardiovascular disorders was collected via electronic search (using Pubmed, SciFinder, Scirus, GoogleScholar, JCCC@INSTIRC and Web of Science) and a library search for articles published in peerreviewed journals. No restrictions regarding the language of publication were imposed. RESULTS: This article highlights the recent scientific findings on four common spices viz. Greater cardamom (Amomum subulatum Roxb.), Coriander (Coriandrum sativum L.), Turmeric (Curcuma longa L.) and Ginger (Zingiber officinale Roscoe), for the role they can play in the management of heart diseases. Although they have been used by many cultures since ancient times and have been known to exhibit several medicinal properties, current research shows that they can also be effectively used for the prevention and control of CVDs. CONCLUSION: Although scientific evidences supporting the benefits of spices in maintaining a healthy heart are available, more complete information is needed about the actual exposures to these dietary components that are required to bring about a response. The innumerable actions of spices that have been shown in in vitro experiments need to be demonstrated in more systematic, well-designed animal model studies. More rigorous clinical trials at the normally consumed levels are needed to determine long-term benefits as well as to assess adverse effects if any at higher concentrations, especially if consumed over longer periods. Once these extensive studies are carried out, it will be easy to define the appropriate intervention strategies utilizing these commonly used spices for achieving the maximum benefits on cardiovascular health without producing any ill-effects.

Determination of flavonoids, polyphenols and antioxidant activity of Tephrosia purpurea: a seasonal study.

OBJECTIVE: Tephrosia purpurea (Linn·) Pers. is widely used in traditional medicine to treat liver disorders, febrile attacks, enlargement and obstruction of liver, spleen, and kidney. In the present study, investigations were carried out to determine the seasonal impact on the content of flavonoid glycosides and on antioxidant activities so as to identify the optimal time of harvesting. METHODS: The plant materials were collected in different seasons during 2013-2014. Air-dried, powdered plant materials were extracted with 95% ethanol and ethanol: water (1:1) by ultrasound-assisted extraction process. Their chemical composition in terms of total polyphenol and flavonoid contents (TPCs and TFCs) was determined using modified colorimetric Folin-Ciocalteu method and aluminum chloride colorimetric assay respectively. To determine the in vitro antioxidant activity, diphenyl-picryl hydrazyl (DPPH) radical-scavenging assay and total antioxidant capacity by phosphomolybdate antioxidant assay were carried out. High-performance liquid chromatography (HPLC)/photo-diode array (PDA) analysis was used to quantify the flavonoid glycosides in the samples collected in different seasons. Correlation studies were also carried out between antioxidant activities and TPCs. RESULTS: The highest TPC and TFC were found to be in the 95% ethanolic extract of the August sample and the lowest in the 50% hydro-alcoholic extract of the plant sample collected in winter season. It was observed that in both the assays used to determine the antioxidant activity, the 95% ethanolic extracts in all the seasons showed a higher activity than their respective 50% hydro-alcoholic extracts with an increase in activity as we go from cold to hot to rainy seasons. Based on correlation analysis, DPPH radical-scavenging activities as well as the spectrophotometrically measured phosphomolybdenum complex were also strongly correlated with TPC of the extracts. The most abundant flavonoid glycoside was quercetin-3-O-rhamnoglucoside in all the seasons. The content of all flavonoids was observed highest in the 95% ethanolic extract of the plant collected in August (TP-3). The 50% hydro-alcoholic extract of the plant collected in December (TP-6) showed the lowest amount of flavonoids and antioxidant activity. CONCLUSION: The findings of this study confirmed that the metabolism and production of flavonoids in T. purpurea are vigorously affected by seasonal factors. Significant differences were observed in the TPC, TFC and flavonoid glycoside composition in the 95% ethanolic as well as 50% hydro-alcoholic extracts of T· purpurea collected in different seasons. Since the 95% ethanolic extract of the August sample (TP-3), which also happens to be its flowering season, exhibited the highest TPC, TFC and antioxidant activities in both DPPH and phosphomolybdate assays as well as contained the highest content of all flavonoids, it could be recommended as the optimal season of harvesting T· purpurea with respect to its pharmaceutically important constituents, i.e., flavonoids.

A Validated HPTLC Densitometric Method for Simultaneous Determination of Evernic and Usnic Acids in Four Usnea Species and Comparison of Their Antioxidant Potential.

A simple, sensitive and precise high-performance thin-layer chromatography method with densitometric detection was used for simultaneous determination of evernic (EV) and (+)-usnic acids (USN) in Usnea aciculifera (UA), U. ghattensis (UG), U. longissima (UL) and U. stigmatoides (US). This method was also validated according to the ICH guidelines. Separation and quantification was performed with the mobile phase toluene-1, 4-dioxane-formic acid (18:4.5:0.2, v/v/v) on silica gel 60F254 plates. The linearity for EV and USN was found in the 200-600 ng/band range. The limit of detection for EV and USN was 51.56 and 32.59 ng/band, while the limit of quantification was 156.23 and 98.76 ng/band, respectively. Intra- and interday precisions (n = 6) for EV and USN were 0.70-1.89 and 0.50-0.76 (%RSD), and 1.56-1.60 and 1.54-1.99 (%RSD), respectively. The mean percent recoveries were 99.66 and 99.87%, respectively, for EV and USN. However, USN was estimated in all four Usnea species but EV only in two species with varied quantity. Comparative antioxidant activity revealed that US is a better free radical scavenger in comparison with other three Usnea species. Furthermore, these results indicated that USN and EV are not solely responsible for antioxidant potential, but it may be due to synergistic effect.

Simultaneous Quantification of Syringic Acid and Kaempferol in Extracts of Bergenia Species Using Validated High-Performance Thin-Layer Chromatographic-Densitometric Method.

A rapid, sensitive, selective and robust quantitative densitometric high-performance thin-layer chromatographic method was developed and validated for separation and quantification of syringic acid (SYA) and kaempferol (KML) in the hydrolyzed extracts of Bergenia ciliata and Bergenia stracheyi. The separation was performed on silica gel 60F254 high-performance thin-layer chromatography plates using toluene : ethyl acetate : formic acid (5 : 4: 1, v/v/v) as the mobile phase. The quantification of SYA and KML was carried out using a densitometric reflection/absorption mode at 290 nm. A dense spot of SYA and KML appeared on the developed plate at a retention factor value of 0.61 ± 0.02 and 0.70 ± 0.01. A precise and accurate quantification was performed using linear regression analysis by plotting the peak area vs concentration 100-600 ng/band (correlation coefficient: r = 0.997, regression coefficient: R(2) = 0.996) for SYA and 100-600 ng/band (correlation coefficient: r = 0.995, regression coefficient: R(2) = 0.991) for KML. The developed method was validated in terms of accuracy, recovery and inter- and intraday study as per International Conference on Harmonisation guidelines. The limit of detection and limit of quantification of SYA and KML were determined, respectively, as 91.63, 142.26 and 277.67, 431.09 ng. The statistical data analysis showed that the method is reproducible and selective for the estimation of SYA and KML in extracts of B. ciliata and B. stracheyi.

Nutritional evaluation, antioxidant studies and quantification of poly phenolics, in Roscoea purpurea tubers.

BACKGROUND: Roscoea purpurea (Zingiberaceae) is commonly known as "kakoli". Traditionally, various parts like leaves, roots and flower etc. are used for the treatment of diabetic, hypertension, diarrhea, fever, inflammation etc. In Nepal tubers are boiled for edible purpose and also used in traditional veterinary medicine. The study aims for nutritional characterization, chemical profiling of R. purpurea (tubers) methanol extract (RPE) along with evaluation of its anti-oxidant activity. Physicochemical and nutritional content were estimated as per standard protocols. Chemical profiling of markers includes method optimization, identification & quantification of bioactive poly phenolics through HPTLC. Anti oxidant potential RPE was analyzed via. Total phenolics (TPC), total flavonoids (TFC), reducing power assay, DPPH and ß-carotene bleaching model. RESULTS: Physicochemical and nutritional standards were established. Kaempferol (0.30%), vanillic acid (0.27%), protocatechuic (0.14%), syringic (0.80%) and ferulic acid (0.05%) were identified and then quantified. TPC and TFC content were found to be 7.10 ± 0.115 and 6.10 ± 0.055%, reducing power of extract also increases linearly (r(2) = 0.946) with concentration, similar to standards. IC50 value of extract in DPPH and ß-carotene bleaching model was observed at 810.66 ± 1.154 and 600.66 ± 1.154 µg/ml, which is significantly different from standards (p < 0.05). Although there is a positive, significant correlation between the phenolic and flavonoid content with anti oxidant activity of extract. CONCLUSION: Thus, study will authenticates the identity, utility of herb as nutrient supplement and an important medicinal plant having promising pharmacological activities for further elaborated/extended investigation work.

Biosynthesis of stable antioxidant ZnO nanoparticles by Pseudomonas aeruginosa rhamnolipids.

During the last several years, various chemical methods have been used for synthesis of a variety of metal nanoparticles. Most of these methods pose severe environmental problems and biological risks; therefore the present study reports a biological route for synthesis of zinc oxide nanoparticles using Pseudomonas aeruginosa rhamnolipids (RLs) (denoted as RL@ZnO) and their antioxidant property. Formation of stable RL@ZnO nanoparticles gave mostly spherical particles with a particle size ranging from 35 to 80 nm. The RL@ZnO nanoparticles were characterized by UV-visible (UV-vis) spectroscopy, scanning electron microscopy, transmission electron microscopy, dynamic light scattering, Fourier transform infrared spectroscopy, X-ray diffraction (XRD), and thermal gravimetric analysis. The UV-vis spectra presented a characteristic absorbance peak at ∼ 360 nm for synthesized RL@ZnO nanoparticles. The XRD spectrum showed that RL@ZnO nanoparticles are crystalline in nature and have typical wurtzite type polycrystals. Antioxidant potential of RL@ZnO nanoparticles was assessed through 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion free radicals with varying concentration and time of the storage up to 15 months, while it was found to decline in bare ZnO nanoparticles. Similarly, the inhibitory effects on ß-carotene oxidation and lipid peroxidation were also observed. These results elucidate the significance of P. aeruginosa RL as effective stabilizing agents to develop surface protective ZnO nanoparticles, which can be used as promising antioxidants in biological system.

Ellagic acid & gallic acid from Lagerstroemia speciosa L. inhibit HIV-1 infection through inhibition of HIV-1 protease & reverse transcriptase activity.

BACKGROUND & OBJECTIVES: Banaba (Lagerstroemia speciosa L.) extracts have been used as traditional medicines and are effective in controlling diabetes and obesity. The aim of this study was to evaluate the anti-HIV property of the extracts prepared from the leaves and stems of banaba, and further purification and characterization of the active components. METHODS: Aqueous and 50 per cent ethanolic extracts were prepared from leaves and stems of banaba and were evaluated for cytotoxicity and anti-HIV activity using in vitro reporter gene based assays. Further, three compounds were isolated from the 50 per cent ethanolic extract of banaba leaves using silica gel column chromatography and characterization done by HPLC, NMR and MS analysis. To delineate the mode of action of the active compounds, reverse transcriptase assay and protease assay were performed using commercially available kits. RESULTS: All the extracts showed a dose dependent inhibition of HIV-1-infection in TZM-bl and CEM-GFP cell lines with a maximum from the 50 per cent ethanolic extract from leaves (IC 50 = 1 to 25 µg/ml). This observation was confirmed by the virus load (p24) estimation in infected CEM-GFP cells when treated with the extracts. Gallic acid showed an inhibition in reverse transcriptase whereas ellagic acid inhibited the HIV-1 protease activity. INTERPRETATION & CONCLUSIONS: The present study shows a novel anti-HIV activity of banaba. The active components responsible for anti-HIV activity were gallic acid and ellagic acid, through inhibition of reverse transcriptase and HIV protease, respectively and hence could be regarded as promising candidates for the development of topical anti-HIV-1 agents.

Extracts from Acacia catechu suppress HIV-1 replication by inhibiting the activities of the viral protease and Tat.

BACKGROUND: Acacia catechu (Mimosa family) stem bark extracts have been used traditionally as a dietary supplement as well as a folk medicine given its reported anti-inflammatory, immunomodulatory, hepatoprotective, antioxidant, anti-microbial and anti-tumor activities. The present study was undertaken to evaluate the anti-HIV-1 activity of the extracts from stem bark of A. catechu. METHODS: The aqueous and 50% ethanolic extracts of A. catechu stem bark were prepared and 50% ethanolic extract was further fractioned by successively partitioning with petroleum ether, chloroform and n-butanol. All the extracts and fractions were evaluated for cytotoxicity and anti-HIV-1 activity using different in vitro assays. The active n-butanol fraction was evaluated for its inhibition against HIV-1 reverse transcriptase, integrase, protease, pro-viral genome integration and viral Tat protein mediated transactivation. The effect of n-butanol fraction on the induction of pro-inflammatory cytokines secretion in Vk2/E6E7 cells and transepithelial resistance in Caco-2 and HEC-1A cells was investigated. RESULTS: The aqueous and 50% ethanolic extracts of A. catechu showed IC50 values of 1.8 ± 0.18 µg/ml and 3.6 ± 0.31 µg/ml, respectively in cell-free virus based assay using TZM-bl cells and HIV-1NL4.3 (X-4 tropic). In the above assay, n-butanol fraction exhibited anti-HIV-1 activity with an IC50 of 1.7 ± 0.12 µg/ml. The n-butanol fraction showed a dose-dependent inhibition against HIV-1NL4.3 infection of the peripheral blood lymphocytes and against HIV-1BaL(R-5-tropic) as well as two different primary viral isolates of HIV-1 infection of TZM-bl cells. The n-butanol fraction demonstrates a potent inhibitory activity against the viral protease (IC50 = 12.9 µg/ml), but not reverse transcriptase or integrase. Further, in Alu-PCR no effect on viral integration was observed. The n-butanol fraction interfered with the Tat-mediated Long Terminal Repeat transactivation in TZM-bl cells, mRNA quantitation (qRT-PCR) and electrophoretic mobility shift assay (EMSA). The n-butanol fraction did not cause an enhanced secretion of pro-inflammatory cytokines in Vk2/E6E7 cells. Additionally, no adverse effects were observed to the monolayer formed by the Caco-2 and HEC-1A epithelial cells. CONCLUSIONS: The results presented here show a potential anti-HIV-1 activity of A. catechu mediated by the inhibition of the functions of the viral protein and Tat.

Acaricidal activity of Cassia alata against Rhipicephalus (Boophilus) annulatus.

Using adult immersion test, the acaricidal activity of ethanolic extracts of leaves of Cassia alata L. was studied against Rhipicephalus (Boophilus) annulatus. The efficacy was assessed by measuring per cent adult mortality, inhibition of fecundity and hatching rate. The ethanolic extract of C. alata produced a concentration dependant increase in the adult tick mortality. The highest mortality (45.8%) and inhibition of fecundity (10.9%) were observed at the highest concentration tested (100 mg/ml). The plant extract did not affect egg hatchability.

Toxic effects of various solvents against Rhipicephalus (Boophilus) annulatus.

The current need of identification of a new acaricidal agent which is acceptable to public as environmentally safe is a daring task. Use of herbal acaricides is such an alternative. Most of the herbal extracts or fractions are dissolved in polar or non-polar solvents or detergents before tested for acaricidal activity. The solvent or detergent to be used for dissolving the herbal extract should be of little acaricidal activity. In the present study, experimentations were carried out on adult engorged female ticks to detect the toxicity of different solvents, viz. hexane, petroleum ether, n-butanol, isopropyl alcohol, chloroform, glycerol, ethyl acetate, acetone, ethanol, and methanol. The study revealed that methanol was the least toxic solvent against Rhipicephalus (Boophilus) annulatus.

An ethnomedicinal, phytochemical and pharmacological profile of Desmodium gangeticum (L.) DC. and Desmodium adscendens (Sw.) DC.

ETHNOPHARMACOLOGICAL RELEVANCE: Desmodium gangeticum (L.) DC. and Desmodium adscendens (Sw.) DC. are two important and well explored species of genus Desmodium (Fabaceae (alt. Leguminosae) subfamily: Faboideae). Desmodium gangeticum is used as a tonic, febrifuge, digestive, anticatarrhal, antiemitic, in inflammatory conditions of chest and in various other inflammatory conditions in the Ayurvedic System of Medicine while Desmodium adscendens is widely used for the treatment of asthma in Ghana, Africa. AIM OF THE REVIEW: The aim of this review is to provide comprehensive information on the botany, traditional uses, phytochemistry, pharmacological research and toxicology of Desmodium gangeticum and Desmodium adscendens to explore their therapeutic potential and future research opportunities. MATERIALS AND METHODS: All the available information on Desmodium gangeticum and Desmodium adscendens was collected via electronic search (using Pubmed, SciFinder, Scirus, Google Scholar, JCCC@INSTIRC and Web of Science) and a library search for articles published in peer-reviewed journals. RESULTS: About 25 different species of Desmodium including Desmodium gangeticum and Desmodium adscendens are used ethnomedicinally all over the world. Phytochemical research on Desmodium gangeticum and Desmodium adscendens has led to the isolation of alkaloids, pterocarpans, phospholipids, sterols, flavones and flavonoid glycosides from Desmodium gangeticum and triterpenoid saponins, phenylethylamines and indole-3-alkyl amines from Desmodium adscendens. Crude extracts, fractions and isolated components of Desmodium gangeticum and Desmodium adscendens showed a wide spectrum of in vitro and in vivo pharmacological activities like antileishmanial, immunomodulatory, antiasthmatic, smooth muscle relaxant, anti-inflammatory, anti-ulcer, cardio-protective, antidiabetic, antiamnesic, antiviral, antioxidant and hepatoprotective activities. CONCLUSIONS: Desmodium gangeticum and Desmodium adscendens have emerged as a good source of traditional medicine. Desmodium gangeticum possesses the ability to scavenge the free radicals generated during ischaemia and ischaemia reperfusion thereby preserving the mitochondrial respiratory enzymes that eventually lead to cardio-protection and has potential prophylactic and therapeutic efficacy against Leishmania infection. Desmodium adscendens is useful against chronic bronchitis and asthma. However, there is a need to search for individual secondary metabolites responsible for these actions and study their mode of actions, bioavailability, pharmacokinetics and physiological pathways in sufficient detail. The promising results should be further substantiated by clinical trials.

Diazinon resistant status in Rhipicephalus (Boophilus) microplus collected from different agro-climatic regions of India.

The resistance status of Rhipicephalus (Boophilus) microplus to "Diazinon" was evaluated in 20 locations situated at various agro-climatic regions of India. Adult immersion test (AIT) was optimized using laboratory reared acaricide susceptible IVRI-I strain of R. (B.) microplus and minimum effective concentration of Diazinon was determined as 635.2 ppm. The discriminating dose (DD) was worked out as 1270.4 ppm and was tested on female ticks collected from organized and unorganized farms located at different agro-climatic regions of India. On the basis of the data generated on three variables viz., mortality, egg masses and reproductive index, the resistance level was categorized as I, II, III and IV. The average resistance factor (RF) of 6.1 (level II) was recorded in the ticks collected from the northern sub-temperate trans-gangetic plains while high average RF values of 26.65 (level III) was recorded in the ticks collected from tropical middle-gangetic plains. The tropical middle gangetic plain has a very high density of animal populations where farmers use Diazinon for tick control, for agricultural practices and for mosquito control. Due to the continuous use of OP compounds the environmental load of Diazinon has become high in the area. This is the first experimental data generated on Diazinon resistant status in ticks of India.