Purpose: The objective of this study was to investigate the functional and molecular expression of a carrier mediated system responsible for folate uptake in breast cancer (BC) (T47D) cells and to delineate the mechanism of intracellular regulation of this transport system. Materials and Methods: [ ³ H]-folic acid uptake was studied in T47D cells with respect to time, pH, temperature, sodium and chloride ion dependency. Inhibition studies were conducted in the presence structural analogs, vitamins, metabolic and membrane transport inhibitors. [ ³ H]-folic acid uptake was also determined with varying concentrations of cold folic acid. Uptake kinetics was studied in the presence of various modulators of intracellular regulatory pathways; calcium-calmodulin, protein kinases A and C (PKA and PKC) and protein tyrosine kinase (PTK). Molecular evidence was studied by qualitative and quantitative polymerase chain reaction (PCR) and Western blot analysis. Results: Linear increase in [ ³ H]-folic acid uptake was observed over 30 min. The process followed saturation kinetics with an apparent K _m of 11.05 nM, V _max of 1.54 * 10⁻⁸ μmoles/min/mg proteins and K _d of 9.71 * 10⁻⁶ /min for folic acid. Uptake process was found to be dependent on pH, sodium ions, chloride ions, temperature and energy. Uptake was inhibited in the presence of structural analogs (cold folic acid, methyltetrahydro folate and methotrexate), but structurally unrelated vitamins did not show any effect. Membrane transport inhibitors such as SITC, DIDS, probenecid and endocytic inhibitor colchicine significantly inhibited the [ ³ H]-folic acid uptake process. PKA, PTK and Ca ²⁺ /calmodulin pathways positively regulate the uptake process. Reverse transcriptase polymerase chain reaction (RT PCR) analysis had shown mRNA expression of folate receptor (FR)-α at 407 bp. Quantitative polymerase chain reaction analysis showed significantly higher FR-α mRNA levels in T47D cells compared to MCF-7 cells and Western blot analysis confirmed the FR-α protein expression at 37 kDa. Conclusions: This work demonstrated the functional characterization and molecular presence of FR-α in the T47D cell line. The high expression of FRs in T47D human breast carcinoma cells supports their validity as molecular therapeutic targets in BC.

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Cichorium intybus L. (chicory) is a Mediterranean plant species belonging to the Asteraceae family. Chicory is gaining interests because of its culinary features, nutritional values and medicinal characteristics. C. intybus has been implemented in folk medicine from North Africa to South Asia for several 100 years. In Indian medicine, it has been used to treat fever, diarrhea, spleen enlargement, jaundice, liver enlargement, gout, and rheumatism. In China, it is valued for its tonic effects upon the liver and digestive tract. In Germany, chicory has been used as a folk medicine for everyday ailments. Thus, C. intybus is a plant of great economic potential due to high concentrations of fructooligosacharide, known as inulin, in its roots, used as a replacement ingredient for sugar and fat. The other various phytoconstituents reported in chicory are sucrose, cellulose, proteins, caffeic acid derivatives, flavonoids, polyphenols, carotenoids, anthocyanins, tannins, coumarins, sesquiterpene lactones, fatty acids, pectin, cholins, benzo-isochromenes, alkaloids, vitamins, amino acids, and minerals. The therapeutic investigations reveal that C. intybus is useful for maintaining normal health and has nematicidal, antihepatotoxic, antidiabetic, cardioprotective, antiallergic, antihyperlipidemic, anti-inflammatory, antineoplastic, calcium homeostater, bulking agent, immunostimulatory, prebiotic, protective against pancreatitis, antimicrobial, and antioxidant effects. This review encompasses botany, ethnomedicinal uses, phytoconstituents, pharmacological uses, and toxicity studies of C. intybus L.

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Background: Euphorbia hirta L. (Euphorbiaceae), commonly known as Dudhani, is distributed in the warm region of India and China. Traditionally, it is used in respiratory and gastrointestinal disorders. In addition, the antidiabetic property of the plant was also reported in the literature. This study was designed to evaluate the effect of aqueous, hydroalcoholic, and methanolic extracts (MEHs) of E. hirta on α-glucosidase and α-amylase in vitro. Materials and Methods: Aqueous, hydroalcoholic, and MEHs of E. hirta were prepared as per application program interface. In α-glucosidase activity, α-glucosidase (0.1 μ/mL) and substrate, 2.5 mM p-nitrophenyl-α-D-glucopyranoside was used; absorbance was recorded at 405 nm. In α-amylase activity, α-amylase solution (1.0 μ/mL) and substrate, 0.25% starch were used, and absorbance was measured at 540 nm. The IC₅₀values were calculated by linear regression. Results: All the extracts showed α-glucosidase inhibitory activity comparable to acarbose with MEH having highest inhibitory activity among tested extracts. The observed IC₅₀values were 213.63, 146.9, 78.88, and 8.07 μg/mL for aqueous, hydroalcoholic, MEH, and acarbose, respectively. All the extracts have shown mild α-amylase inhibitory activity compared to acarbose. Lineweaver–Burk plot has shown that the MEH is a mixed noncompetitive inhibitor for α-glucosidase enzyme. Conclusion: The results from this in vitro study clearly indicated that MEH of E. hirta had strong inhibitory activity against α-glucosidase and mild inhibitory activity against α-amylase. It can be used for management of postprandial hyperglycemia with lesser side effects, and provide a strong rationale for further animal and clinical studies.

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Purpose: The objective of this study was to develop a novel peptide prodrug of doxorubicin which can evade over-expressed efflux pumps in breast cancer cells. This approach may lead to increased uptake and higher drug accumulation in nuclei of cancer cells. Materials and Methods: L-val-L-val doxorubicin prodrug was synthesized following standard f-moc chemistry. The prodrug was analyzed for stability, cellular and nuclear uptake and interaction with efflux and peptide transporters. Breast cancer cells (T47D) were grown on polystyrene 12-well plates. Result: The prodrug Val-Val-doxorubicin was found to be very stable in breast cancer cell homogenate. It was able to evade efflux pumps. The prodrug penetrated cytoplasm and nucleus of cancer cells by interacting with peptide transporters.These transporters (pepT1 and pepT2) are expressed both on plasma and nuclear membrane of breast cancer cells. Uptake of prodrug was found to be 10 times more than parent drug. Conclusion: Peptide prodrug derivatization of doxorubicin has potential to evade efflux pumps and increase availability and nuclear accumulation of doxorubcin in breast cancer cells.

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Introduction: This study investigated the hepatoprotective activity of aqueous root bark extracts of Anogeissus leiocarpus, Terminalia avicennioides, and their 1:1 mixture in diethylnitrosamine (DEN)-treated rats. Materials and Methods: Rats were orally pretreated with extracts and 1:1 mixture (200, 400, and 800 mg/kg body weight) for 2 weeks before being challenged with DEN (30 mg/kg body weight) once a week on weeks 3 and 4. Curcumin (200 mg/kg body weight) was used as reference antioxidant agent, and distilled water was used as a control. The whole experiment lasted for 4 weeks. The activities of liver and serum alkaline phosphatase (ALP), aspartate and alanine aminotransferases (AST and ALT), and gamma-glutamyltransferase (GGT), as well as the levels of serum total protein, albumin, total, and conjugated bilirubin were evaluated. Results: The extracts and their mixture significantly (P < 0.05) prevented the DEN-induced alterations in the activities of ALP, AST, ALT, and GGT in the liver and serum of the animals; as well as the altered levels of serum total and conjugated bilirubin. Conclusion: Overall, the results of this study revealed that the extracts possess hepatoprotective activities against DEN-induced liver damage in rats.

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Background and Objectives: Typhoid fever is endemic in India. Extensive use of first-line antibiotics has led to the emergence of multi-drug resistant (MDR) Salmonella typhi. Ciprofloxacin has become empirical therapy of choice against MDR salmonellae. Recent year's emergence of low-level ciprofloxacin resistance in salmonellae resulted in delayed response and serious complications. Nalidixic acid (NA) screen test is used as surrogate marker for detection low-level ciprofloxacin resistance. In this study, we evaluated prevalence of MDR and low-level ciprofloxacin resistant S. typhi and Salmonella paratyphi A. Materials and Methods: A total of 50 blood culture isolates of S. typhi and S. paratyphi A were tested for antibiotic susceptibility according to Clinical Laboratory Standards Institute (CLSI) method. Minimal inhibitory concentration (MIC) to ciprofloxacin was carried out by E-test and agar dilution method. Results: Among the 50 salmonella isolates, 80% were S. typhi and 20% were S. paratyphi A. MDR was found in 2% S. typhi. NA resistant salmonellae showed ciprofloxacin MIC ranging from 0.25 to 0.75 μg/ml. One isolate of S. typhi showed ciprofloxacin MIC of 32 μg/ml and was also resistant to ceftriaxone. NA screen test for low-level ciprofloxacin resistance was 100% sensitive and 97.9% specific. Interpretation and Conclusion: NA resistant isolates should be tested for ciprofloxacin MIC to decide therapeutic options. The current CLSI breakpoints may have to be re-evaluated for salmonellae.

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Introduction: Glucosamine-6-phosphate (GlcN6P) synthase biosynthetic pathway has been identified as potential targets for the development of new antimicrobial agents. Aim: A series of 2-chloro-N-(42-phenylthiazol-25-yl)acetamide derivatives (3a-r) was synthesized and evaluated their antimicrobial activity. Materials and Methods: The 2-chloro-N-(Para substituted phenylthiazol-25-yl) acetamide (2a-c) were synthesized by stirring intermediates (1a-c) with 2-chloroacetylchloride in dichloromethane in the presence of K2CO3. The intermediate (2a-c) were further reacted with different secondary amine such as pyrrolidine, N-methyl piperazine, N-ethyl piperazine, thiomorpholine, morpholine, piperidine etc in ethanol in presence of TEA Triethylamine (TEA) to get desired compounds (3a-r). Compounds were characterized by a spectroscopic technique such as Fourier transform infraredFTIR, ¹ H-NMR, ¹³ C-NMR, and mass spectrometry. The synthesized thiazole derivatives (3a-r) were screened for anti-bacterial and anti-fungal activity against Escherichia coli, Staphylococcus aureus NCTC 6571, Pseudomonas aeruginosa NCTC 10662, CandidaC. albicans (MTCC-183), AspergillusA. niger (MTCC 281) NCTC 10418 and AspergillusA. flavus (MTCC 277). Result and Conclusion: The results of anti-bacterial screening revealed that among all the screened compounds, eight compounds viz. 3b, 3c, 3d, 3e, 3i, 3j, 3k, and 3p showed moderate to good anti-bacterial and antifungal activity having minimum inhibitory concentration (MIC) between 6.25- and 25 µg/ml. While compound 3d showed the most promising antibacterial activity against E. coli and S. aureus, while the compound 3j showed promising antifungal activity with MIC value 6.25 µg/ml against C. albicans, A. niger and A. flavus. In addition, all these eight potential molecules were also examined for possible binding on enzyme GlcN6Pglucosamine-6-phosphate synthase by molecular docking studies on (PDB ID 1JXA).

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Background: Zonisamide (ZON) and amlodipine (AML) were studied in different experimental models of epilepsy individually. However, combination of ZON and AML has not been explored yet. Hence, this study was aimed to evaluate the combination therapy of ZON and AML on pentylenetetrazol (PTZ)-induced kindling in mice. Materials and Methods: Swiss albino mice of either sex were randomly divided into five groups and each group containing 8 mice. Kindling was induced by PTZ (45 mg/kg/day on 8 ^th , 10 ^th and 12 ^th and 75 mg/kg/day on 14 ^th day). After treatment animals were observed for 30 min for behavioral analysis then animals were sacrificed and brain was taken out for biochemical analysis. Results: PTZ-significantly induced seizure was characterized by alteration in seizure score and latency as well as significantly increased in brain thiobarbituric acid reactive substance (TBARS) levels and significantly decreased in reduced glutathione, catalase (CAT) and superoxide dismutase (SOD). Treatment with ZON and AML significantly (P < 0.05, P < 0.01 and P < 0.001) resorted seizure score, latency, TBARS, reduced glutathione, CAT and SOD near to normal compared with pentylentetrazol treated rats. Conclusion: This study provides experimental evidence that treatment with both ZON and AML attenuated seizure and oxidative stress in PTZ-induced kindling mice.

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Background: The present study was designed to develop an efficient protocol for studying the enhancement of in vitro production of secondary metabolites in Adhatoda vasica leaves collected from the varied geographical locations of Indian sub-continents by a validated simultaneous high-performance liquid chromatography gradient method for the quantification of vasicine and vasicinone in the developed calli and crude samples. Materials and Methods: The analysis was carried out on a reverse phase C₁₈column using 0.1% of orthophosphoric acid: Acetonitrile in gradient manner and carried out the detection at 280 nm wavelength keeping the flow rate of 1.0 mL/min. Results: The simultaneous method was found linear with regression coefficient r² = 0.991 in a wide range (100–500 μg/mL) precise, accurate, and robust for quinazoline alkaloids in samples. Results clearly showed a significant increase in the concentration of alkaloids in in vitro cultures as compared to natural ones. The proposed method was validated as per the International Conference on Harmonization guidelines for accuracy, precision, robustness, limit of detection, and limit of quantitation. Conclusion: The developed method was found suitable for quality control of A. vasica and for the analysis of vasicine and vasicinone in any type of sample.

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Background: Most of the herbal medicines in the world originate from the developing countries. There are ample opportunities for these countries to expand their global export. The world market for botanical medicines including drug products and raw materials has been estimated to have an annual growth rate between 5% and 15%. Total global botanical drug market is estimated at US$62 billion and is expected to grow to the tune of US$5 trillion by the year 2050. In the USA alone, the usage of botanicals has been increased by 380% between the years 1990 and 1997. Materials and Methods: Ayurveda, the Indian system of medicine, is one of the ancient, yet living traditions that face a typical Western bias. Widespread and growing use of botanicals has created public health challenges globally in terms of quality, safety, and efficacy. Results and Discussion: The development of parameters for standardization and quality control of botanicals is a challenging task. Various regulatory authorities, research organizations, and botanical drug manufacturers have contributed in developing guiding principles and addressing issues related to the quality, safety, and efficacy. Conclusions: The present review describes the regulatory aspects of herbal drugs in India and various other countries.

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Background: Contamination with aflatoxin, by Aspergillus flavus, is one the major challenges in agriculture and food industry. Preparation of organic products using natural components is widely considered these days. Aims: In this study, effects of aqueous extracts of five medicinal herbs, including thyme, senna, mentha, basil, and safflower on the growth of the A. flavus were investigated. Mterials and Methods: The extracts with different concentrations (200-800 µg/mL) and polyethylene glycol with the equal osmotic potential of plant extracts were added to the potato dextrose agar medium to evaluate fungus growth after 7 days using agar dilution method. Benomyl, a fungicide, was used as a positive standard. The tests were performed in triplicate, and the mean diameters of fungus growth were calculated as well. Results and Conclusion: All concentrations of the plants extracts significantly inhibited the fungus growth in comparison with each other and control treatments, while the extracts of thyme and safflower manifested the most effective prohibition compared to benomyl with minimum inhibitory concentration of 200 and 400 µg/mL, respectively.

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Background: Ritonavir (RTV) and atazanavir sulfate (ATV) are protease inhibitor and RTV mostly used as a booster for increasing the bioavailability of other protease inhibitors like ATV. Aims: Quality assessment of the new dosage form of RTV and ATV i.e., tablets is very essential and hence this work deals with to develop sensitive, simple and precise method for simultaneous estimation of ATV and RTV in tablet dosage form by absorbance correction method. Materials and Methods: The present work was carried out on Shimadzu Ultraviolate(UV)-1700 double beam spectrophotometer with 1 cm path length supported by S Shimadzu, model-1700(Japan), UV-Probe software, version 2.31 was used for spectral measurements with 10 mm matched quartz cells. Standard ATV and RTV were supplied by Cipla Pharmaceutical Ltd. Methanol was purchased from Finar Chemicals Pvt. Ltd. Results and Conclusion: The λ_max or the absorption maxima for ATV and RTV were found to be 279 and 240 nm, respectively in methanol as solvent. The drugs follow Beer-Lambert's law in the concentration range 30-90 and 10-30 μg/mL for ATV and RTV, respectively. The percentage recovery was found to be 100-100.33% and 100-101.5% for ATV and RTV, respectively. The method was validated for different parameters as per the International Conference for Harmonization Guidelines.

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Background: Herbs basically, are plants or plant parts employed due to their scent, medicinal properties or flavor. Herbal medicines have longed been used in the management and treatment of various ailments even before the arrival of modern medicine. Herbal medicines are still being used today, as it has even gained a new momentum in the field of medicine. Phytochemicals are responsible for eliciting definite physiological effects of various herbs in the body. In Nigeria, various plants are being used traditionally in the treatment of divers ailments. Some of these plants include Tapinanthus bangwensis, Tamarindus indica, Ocimum gratissimum, Allium sativum, Kigelia africana, Azadirachta indica, Solanum virginianum, Myrianthus serratus and Vernonia amygdalina. Though there have been claims of success in their traditional usage, it is however important to carry-out phytochemical assessment on them. Aim: The aim was to evaluate the phytochemical constituents of the ethanolic extract of these plants. Materials and Methods: The plant materials were extracted, and the plant extracts were screened for the presence of various phytochemical constituents such as alkaloids, steroids, glycosides, cardiac glycosides, flavonoids, carbohydrate, saponins, tannins and anthraquinones using the standard methods. Result: The experimental result revealed the presence of the various bioactive phytochemicals in the different plant extracts investigated. These phytochemicals were however present in different proportions in the various plant extracts. Conclusion: The experimental result vindicates the usage of these plants traditionally for medicinal purposes.

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Aim and Objectives: A simple, rapid, and sensitive high performance liquid chromatographic method with ultraviolet detection has been developed and validated according to the International Conference on Harmonization (ICH) guidelines for the quantitation and qualification of zidovudine (ZID), lamivudine (LAM), and nevirapine (NEV) in pharmaceutical dosage forms. Materials and Methods: The proposed method was based on the separation of the drugs in reversed phase mode using Water's C18 250 cm × 4.6 mm, 5 μ particle size column maintained at an ambient temperature. The optimum mobile phase consisted of Water: Methanol (70:30 v/v), pH adjusted to four with orthophosphoric acid (OPA). The flow rate of mobile phase was set 1.0 mL min ^-1 and photodiode array detection was performed at 275 nm with a total run time of 8 min which is very short for accurate analysis of simultaneous estimation of three analytes. The method was validated according to ICH guidelines. Results: The method was linear over the concentration range of 25-75 μg mL ^-1 with limit of quantifications (LOQ) of 13, 0.49, and 0.40 ng mL ^-1 for ZID, LAM and NEV respectively and limit of detection (LOD) of 4, 0.14, and 0.12 ng mL ^-1 for ZID, LAM and NEV respectively. Accuracy (% recovery studies) and precision values of both inter and intraday obtained from six different replicates for all the analytes ranged from 99.00% to 100.00% and % relative standard deviation of precision (assay) was between 0.64 and 1.28, respectively. All the three analytes and their combination drug product were exposed to thermal, photolytic, hydrolytic, reductive, oxidative and peroxide stress conditions and the stressed samples were analyzed by the proposed method. There were no interfering peaks from excipients, impurities or degradation products due to variable stress conditions and the proposed method is specific for the simultaneous estimation of ZID, LAM and NEV in the presence of their degradation products. Conclusion: The proposed method can be successfully applied in the quality control and stability samples of pharmaceutical dosage forms.

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Background: The aim of the present research work is to develop a high-performance liquid chromatography (HPLC) method for simultaneous analysis of 18α-glycyrrhetinic acid (18α-GA) and 18β-GA (18β-GA) of Glycyrrhiza glabra. Materials and Methods: About 20 μL aliquots of each 18α-GA and 18β-GA were analyzed using reversed-phase C-18 column. The mobile phase was acetonitrile:tetrahydrofuran:water (10:80:10, v/v/v). The run time was 10 min at flow rate of 1 ml/min. Ultraviolet detection was carried out at 254 nm. Results: 18α-GA and 18β-GA were well resolved in reversed phase C-18 column using mobile phase acetonitrile: tetrahydrofuran: water (10:80:10, v/v/v, pH 7.9). The R_tof 18α-GA and 18β-GA was detected at 2.091 and 2.377 min, respectively. Conclusion: The developed chromatography method could be extended for potential quantification or simultaneous determination of these markers in plant as well as in herbal formulation.

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Objective: The aim of the present study is to evaluate and optimize different carbon sources at different temperatures in shake flask for cell growth and astaxanthin accumulation in ultraviolet (UV) mutant Phaffia rhodozyma. The focus is to make the process cost-effective and fruitful with higher productivity. Materials and Methods: Adaptive strain development was carried out so that the P. rhodozyma can give the best astaxanthin at a higher temperature (35°C), increasing the process economy. P. rhodozyma was cultivated with three broad spectrum antibacterial drugs - streptomycin, gentamycin, and ampicillin (200 μL) and with antifungal drug - fluconazole (200 μL) to determine the effect on yeast growth and astaxanthin production. Rice, cane juice, and sucrose were selected as carbon source and compared with dextrose at different fermentation temperatures - 22°C, 30°C, 33°C, and 35°C for astaxanthin production. Results: P. rhodozyma was resistant to antibiotic drugs inhibiting bacterial and fungal contaminants. Rice being more economical was selected as carbon source, but the strain was not able to utilize starch-rich substrate leading to its rejection. When P. rhodozyma was grown in cane juice, biomass is highest (OD 2.77) at 30°C and 610 nm wavelength, whereas astaxanthin productivity is highest (OD 2.64) in dextrose media at 30°C and 450 nm. Comparative metabolic and growth results for UV mutants showed significantly higher biomass and astaxanthin productivity in comparison to wild strain. Conclusion: The most efficient carbon source in terms of penny-pinching and astaxanthin productivity was found to be dextrose containing media at 30°C.

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Objective: This study evaluated the antimicrobial activity of chloroform (CHCl₃) extracts of chyawanprash (Dabur India Ltd., Uttar Pradesh, India) and hydrolyzed chyawanprash against Escherichia coli and Staphylococcus aureus on nutrient agar media using cup plate method. Materials and Methods: CHCl₃extracts of chyawanprash were prepared directly as well as by previously hydrolyzing with dilute HCl. Three test groups of 10, 20, and 50 mg/ml were prepared for each CHCl₃extract, whereas ampicillin 20 mg/ml was used as standard group. Further, six agar plates were prepared for each group, i.e., three for E. coli and three for S. aureus. Cups were bored on each plate with the help of steel cup borer and cups on each plate were marked for identification. Extracts of different concentrations (i.e., 10, 20, and 50 mg/ml) were poured into cups and all plates were then incubated at 37°C for 48 h. After incubation, plates were observed for bacterial growth and zone of inhibition were measured. Results: Results showed that both CHCl₃as well as hydrolyzed CHCl₃extract of chyawanprash showed concentration-dependent antimicrobial activity. The largest zone of inhibition was observed at the concentration of 50 mg/ml of both CHCl₃extracts of chyawanprash (12.7 ± 1.5 mm for E. coli and 15.0 ± 1.0 mm for S. aureus) and hydrolyzed chyawanprash (14.3 ± 0.6 mm for E. coli and 16.3 ± 0.6 mm for S. aureus). Conclusion: Chyawanprash possesses promising potential for use as an antimicrobial agent.

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