Following four foliar applications of Nativo 75 WG (trifloxystrobin 25%+ tebuconazole 50%) on grapes @ 175 and 350 g/ha, resulting in active applications of trifloxystrobin @ 43.75 and 87.5 g a.i./ha and that of tebuconazole @ 87.5 and 175 g a.i./ha, the average initial deposits of trifloxystrobin were observed to be 7.76 and 15.53 mg/kg, respectively, at single and double the application rates. These residue levels dissipated to >85% after 10 days of its application at both the dosages. Similarly, the average initial deposits of tebuconazole were observed to be 13.84 and 26.55 mg/kg at single and double the application rate, respectively. These residue levels dissipated to >90% after 10 days of application at both the dosages. The half-life (t (1/2)) periods of trifloxystrobin on grape leaves were observed to be 2.92 and 3.48 days at single and double application rates, respectively, whereas these values were 2.68 and 3.96 days for tebuconazole. Ripe grape berries and soil samples collected at harvest which happened to be 34 days after the last application, did not show the presence of residues of trifloxystrobin and tebuconazole at their determination limit of 0.01 and 0.02 mg/kg, respectively.

BACKGROUND: A significant fraction of vascular smooth muscle cells (VSMCs) undergo rapid apoptosis after balloon angioplasty. In this study, we tested the hypothesis that protecting VSMCs from undergoing apoptosis prevents the cascade of events that lead to intimal hyperplasia. METHODS AND RESULTS: Rapamycin-loaded gel-like nanoparticles (mean diameter, 54+/-5 nm) were infused locally in a rat carotid artery model of vascular injury. The drug has both antiapoptotic and antiproliferative effects on VSMCs and hence was selected for the current study. Localized delivery of nanoparticles sustained the drug level in the target artery for >2 weeks; demonstrated significant inhibition of hyperplasia (intima/media ratio, 1.5+/-0.02 versus 2.7+/-0.6; P<0.01); and most importantly, reendothelialized the injured artery (endothelium coverage: treated 82% versus control 28%). We also demonstrated inhibition of activation of caspase-3/7 enzymes in the treated artery, preventing VSMCs from undergoing apoptosis and subsequent infiltration of macrophages. CONCLUSIONS: It may be postulated that the localized delivery of rapamycin inhibited apoptosis of VSMCs, minimizing the inflammatory response to the injury and, thus, creating conditions conducive to vascular repair (reendothelialization). Unlike stenting, which can lead to thrombosis and increased risk for in-stent restenosis, our approach could eliminate or minimize long-term complications because the injured artery undergoes a natural process of reendothelialization.

Doxorubicin (DOX) is an anticancer drug with an intracellular site of action in the nucleus. For high antitumour activity, it should be effectively internalized into the cancer cells and accumulate in the nucleus. In this study, we have prepared a nuclear localization signal conjugated doxorubicin loaded Poly (D, L-lactide-co-glycolide) nanoparticles (NPs), to deliver doxorubicin to the nucleus efficiently. Physico-chemical characterization of these NPs showed that the drug is molecularly dispersed in spherical and smooth surfaced nanoparticles. NPs ( approximately 226nm in diameter, 46% encapsulation efficiency) under in vitro conditions exhibited sustained release of the encapsulated drug (63% release in 60 days). Cell cytotoxicity results showed that NLS conjugated NPs exhibited comparatively lower IC(50) value (2.3muM/ml) than drug in solution (17.6muM/ml) and unconjugated NPs (7.9muM/ml) in breast cancer cell line MCF-7 as studied by MTT assay. Cellular uptake studies by confocal laser scanning microscopy (CLSM) and fluorescence spectrophotometer showed that greater amount of drug is targeted to the nucleus with NLS conjugated NPs as compared to drug in solution or unconjugated NPs. Flow cytometry experiments results showed that NLS conjugated NPs are showing greater cell cycle (G2/M phase) blocking and apoptosis than native DOX and unconjugated NPs. In conclusion, these results suggested that NLS conjugated doxorubicin loaded NPs could be potentially useful as novel drug delivery system for breast cancer therapy.

Immunogenicity of different antigen preparations of outer membrane protein (OMP) of Aeromonas hydrophila such as Poly D, L-lactide-co-glycolic acid (PLGA) microparticles, oil emulsion, neat OMP and bacterial whole cells were compared through intra-peritoneal injection in fish, Labeo rohita. Among these preparations, PLGA encapsulated antigen stimulated both innate and adaptive immune parameters and the immunogenicity exhibited by PLGA microparticles was significantly higher (p< 0.05) at both 21 and 42 days post-immunization suggesting that the above delivery system would be a novel antigen carrier for parenteral immunization in fish, Labeo rohita.

Aim: In the development of controlled-release protein therapeutics, the high encapsulation of proteins into biodegradable nanoparticles with uniform size in an anhydrous process along with an excellent redispersion is of practical interest. The objective of this work was to study the physicochemical and in vitro release properties of chitosan nanoparticles with different molecular weights (low, medium and high) using bovine serum albumin (BSA) as a model protein for developing nanoparticle formulations that were stable and reproducible after lyophilization. Materials & methods: The BSA-loaded chitosan nanoparticles were prepared by an ionic gelation method using pentasodium tripolyphosphate as the polyanions. The physicochemical properties and in vitro release kinetics of the nanoparticles were evaluated along with Fourier transform infrared spectroscopy studies. Furthermore, the nanoparticles were freeze-dried for long-term stability in the formulation. To optimize the size of the freeze-dried nanoparticles after redispersion, various types of lyoprotectants (natural and synthetic) were tested in varying concentration in the process of lyophilization. Results: The dynamic light scattering measurements revealed the increase in size of chitosan nanoparticles with the increase in molecular weight of chitosan with no significant change, irrespective of the concentration of BSA entrapped. In addition, the entrapment efficiency of the nanoparticles increased with the increasing molecular weight of chitosan and BSA concentration. By contrast, the redispersity of the freeze-dried samples resulted in further increase of the mean diameter of the nanoparticles. Conclusion: Among the various types of lyoprotectants (natural and synthetic) examined, sucrose proved to be very effective in reducing the size of freeze-dried nanoparticles on redispersion without significant change in surface charge of nanoparticles. Finally, the in vitro release kinetics of BSA from nanoparticles of different molecular weights of chitosan, with and without sucrose, was evaluated and found to depend upon the molecular weight of chitosan.

Calumenin is a multiple EF-hand Ca(2+) binding protein localized in the sarcoplasmic reticulum (SR) with C-terminal SR retention signal HDEF. Recently, we showed evidence that calumenin interacts with SERCA2 in rat cardiac SR (1). The present study was undertaken to further characterize the association of calumenin with SERCA2 in mouse heart by various gene manipulation approaches. Immunocytochemical analysis showed that calumenin and SERCA2 were partially co-localized in HL-1 cells. Knockdown (KD) of calumenin was conducted in HL-1 cells and 80% reduction of calumenin did not induce any expressional changes of other Ca(2+)-cycling proteins. But it enhanced Ca(2+) transient amplitude and showed shortened time to reach peak and decreased time to reach 50% of baseline. Oxalate-supported Ca(2+) uptake showed increased Ca(2+) sensitivity of SERCA2 in calumenin KD HL-1 cells. Calumenin and SERCA2 interaction was significantly lower in the presence of thapsigargin, vanadate or ATP, as compared with 1.3 microM Ca(2+), suggesting that the interaction is favored in E1 state of SERCA2. GST-pull down assay of calumenin deletion fragments and SERCA2 luminal domains suggested that 132-222 amino acids region of calumenin and SERCA2-L4 (853-892 amino acids) are the major binding partners. On the basis of our in vitro binding data and available information on 3D structure of Ca(2+)-ATPases, a molecular model was proposed for the interaction between calumenin and SERCA2. Taken together, the present results suggest that calumenin is a novel regulator of SERCA2 and its expressional changes are tightly coupled with Ca(2+)-cycling of cardiomyocytes.

We have for the first time developed two ligand-assisted Cu(I)-catalyzed sequential intra- and intermolecular S-arylations leading to the direct synthesis of arylthiobenzothiazoles in one pot without an inert atmosphere. Low catalyst loading, inexpensive metal catalyst and ligand, lower reaction temperature, and shorter reaction times make this method superior to all reported methods for the synthesis of arylthiobenzothiazole.

Two novel tripodal imine-phenol ligands, cis,cis-1,3,5-tris{(2-hydroxybenzilidene)aminomethyl}cyclohexane (TMACHSAL, L(1)) and of cis,cis-1,3,5-tris{[(2-hydroxyphenyl)ethylidene]aminomethyl}cyclohexane (Me(3)-TMACHSAL, L(2)) have been synthesized and characterized by elemental analyses and various spectral (UV-vis, IR and (1)H and (13)C NMR) data. The complexation reactions of the ligands with H(+) and Fe(III) were investigated by potentiometric and spectrophotometric methods at an ionic strength of 0.1M KCl and 25+/-1 degrees C in aqueous medium. Three protonation constants each for ligands L(1) and L(2) were determined and were used as input data to evaluate the formation constants of the metal complexes. Formations of metal complexes of the types FeLH(3), FeLH(2), FeLH, FeL and FeLH(-1) were depicted in solution. Experimental evidences suggested for a formation of tris(iminophenolate) type metal complex by the ligands. The ligand L(1) showed higher affinity towards iron(III) than L(2). The pFe value related to L(1)(pFe=20.14) is approximately four units higher than L(2)(pFe=16.41) at pH=7.4. The structures of the metal complexes were proposed through the molecular mechanics calculation using MM3 force field followed by semi-empirical PM3 method.

Exposure due to natural radiation is of particular importance because it accounts for the largest contribution (nearly 85 %) to the total collective dose of the world population. An attempt has been made to present the feasibility of uranium occurrence in drinking water samples from different states of India, by laser-induced fluorimetry. The associated age-dependent radiation dose was estimated by taking the prescribed water intake values of different age groups. The concentration of uranium obtained, i.e. 0.1 +/- 0.01 to 19.6 +/- 1.8 microg l(-1), is well below the drinking water guideline value of 30 microg l(-1). The annual ingestion dose due to uranium in drinking water for various age groups is found to vary from 0.14 to 48 microSv y(-1).

Selective drug delivery is an important approach with great potential for overcoming problems associated with the systemic toxicity and poor bioavailability of antineoplastic drugs. Nanomedicine plays a pivotal role by delivering drugs in a targeted manner to the malignant tumor cells thereby reducing the systemic toxicity of the anticancer drugs. The objective of this study was to prepare and characterize rapamycin loaded polymeric poly(lactide-co-glycolide)(PLGA) nanoparticles (NP) that were surface conjugated with antibodies to epidermal growth factor receptor (EGFR), highly expressed on breast cancer cells, using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) mediated cross linking agents. To potentiate the anticancer efficiency of the formulations, in vitro cytotoxicity of native rapamycin, rapamycin loaded nanoparticles and EGFR antibody conjugated rapamycin loaded nanoparticles (EGFR-Rapa-NPs) were evaluated on malignant MCF 7 breast cancer cell lines. IC(50) doses as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) assay showed the superior antiproliferative activity of EGFR-Rapa-NPs over unconjugated nanoparticles and native rapamycin due to higher cellular uptake on malignant breast cancer cells. Cell cycle arrest and cellular apoptosis induced by the above formulations were confirmed by flow cytometry. Molecular basis of apoptosis studied by western blotting revealed the involvement of a cytoplasmic protein in activating the programmed cell death pathway. Thus it was concluded that EGFR-Rapa-NPs provide an efficient and targeted delivery of anticancer drugs, presenting a promising active targeting carrier for tumor selective therapeutic treatment in near future.