Multifunctional HER2-antibody conjugated polymeric nanocarrier-based drug delivery system for multi-drug-resistant breast cancer therapy.

Nanotechnology-based medical approaches have made tremendous potential for enhancing the treatment efficacy with minimal doses of chemotherapeutic drugs against cancer. In this study, using tamoxifen (Tam), biodegradable antibody conjugated polymeric nanoparticles (NPs) was developed to achieve targeted delivery as well as sustained release of the drug against breast cancer cells. Poly(D,L-lactic-co-glycolic acid) (PLGA) NPs were stabilized by coating with poly(vinyl alcohol) (PVA), and copolymer polyvinyl-pyrrolidone (PVP) was used to conjugate herceptin (antibody) with PLGA NPs for promoting the site-specific intracellular delivery of Tam against HER2 receptor overexpressed breast cancer (MCF-7) cells. The Tam-loaded PVP-PLGA NPs and herceptin-conjugated Tam-loaded PVP-PLGA NPs were characterized in terms of morphology, size, surface charge, and structural chemistry by dynamic light scattering (DLS), Transmission electron microscopy (TEM), ζ potential analysis, 1H nuclear magnetic resonance (NMR), and Fourier transform infrared (FT-IR) spectroscopy. pH-based drug release property and the anticancer activity (in vitro and in vivo models) of the herceptin conjugated polymeric NPs were evaluated by flow cytometry and confocal image analysis. Besides, the extent of cellular uptake of drug via HER2 receptor-mediated endocytosis by herceptin-conjugated Tam-loaded PVP-PLGA NPs was examined. Furthermore, the possible signaling pathway of apoptotic induction in MCF-7 cells was explored by Western blotting, and it was demonstrated that drug-loaded PLGA NPs were capable of inducing apoptosis in a caspase-dependent manner. Hence, this nanocarrier drug delivery system (DDS) not only actively targets a multidrug-resistance (MDR) associated phenotype (HER2 receptor overexpression) but also improves therapeutic efficiency by enhancing the cancer cell targeted delivery and sustained release of therapeutic agents.

Leaf plasticity to light intensity in Italian cypress (Cupressus sempervirens L.): adaptability of a Mediterranean conifer cultivated in the Alps.

Italian cypress (Cupressus sempervirens L.) is native to the eastern Mediterranean, an area characterised by hot, dry summers and mild winters. Over the centuries, however, the species has been introduced into more northerly regions, a long way from its native range. The current, generally warmer climatic conditions brought about by global warming have favoured its cultivation in even more northerly areas in the Alps and other European alpine regions. Given that not only temperature, but also light availability are limiting factors for the spread of cypress in these environments, it is important to ascertain how this species copes with low light conditions. The photosynthetic characteristics of cypress leaves collected from different portions of the crown with contrasting light availability were evaluated by several methods. Chlorophyll a (Chl a), chlorophyll b (Chl b) and carotenoid (Car) content was found to be higher in shade leaves than in sun leaves when measured on a fresh mass basis, although enzymatic activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) and nitrate reductase were lower in shade leaves. When the efficiency of PSII was measured by chlorophyll fluorescence, a marked reduction in F(m) was found in shade leaves, while F(o) remained unchanged. The use of exogenous electron donors diphenyl carbazide (DPC) and NH(2)OH actually improved the photosynthetic efficiency of shade leaves, and the same effect was found when PSII electron transport activity was measured as O(2) evolution. Altogether, these results seem to indicate lesser photosynthetic efficiency in shade leaves, probably an impairment on the donor side of the PSII. At the same time, analysis by SDS-PAGE revealed differences in the polypeptide composition of the thylakoid membranes of sun and shade leaves: the bands corresponding to 23 kDa, 28-25 kDa and 33 kDa polypeptides were less intense in the thylakoid membranes extracted from shade leaves. These results were further confirmed by an immunological study showing that the content of the 33 kDa protein, corresponding to the extrinsic PSII protein PsbO, was significantly diminished in shade leaves. The high plasticity of cypress leaves appears to be an advantageous trait in the plant's response to variations in environmental conditions, including global change. Implications for the management of this Mediterranean species at the northern edge of its distribution are discussed.

Development of in vitro gene delivery system using ORMOSIL nanoparticle: Analysis of p53 gene expression in cultured breast cancer cell (MCF-7).

This article reports on the application of organically modified silica (ORMOSIL) nanoparticles as an efficient in vitro gene delivery system in the recent years. Based on that prime objective, the present study addresses the possible ways to reduce cancers incidence at cellular level. In this context, ORMOSIL nanoparticles had been synthesized and incubated along with pCMV-Myc (3.8 kb) plasmid vector construct carrying p53gene, and transfected into the breast cancer cell line MCF-7 cells. Western blot analysis showed that the p53 protein was significantly expressed in breast cancer cell upon transfection. The confocal and electron microscopic studies further confirmed that the nanoparticles were accumulated in the cytoplasm and the nucleus of the cancer cells transfected with p53 gene. Interesting agarose gel electrophoresis studies revealed that the nanoparticles efficiently complex with pCMV-Myc vector. The anti-cancer properties of p53 were demonstrated by assessing the cell survival and growth rate which showed a positive linear correlation in cancer cells. Whereas, the growth rate was significantly reduced in ORMOSIL/p53/pCMV-Myc transfected breast cancer cells compared to the growth rate of non-transfected cells. The results of this approach using ORMOSIL nanoparticles as a non-viral gene delivery platform have a promising future for use as effective transfection agent for therapeutic manipulation of cancer cells and targeted cancer gene therapy in vivo.

Protective effects of ethanolic extract residue isolated from the bark of Terminalia Arjuna against DLA tumour cells.

An ethanolic herbal extract residue was prepared from the seeds of premature fruits of Terminalia arjuna Linn. Different concentrations of it were tested against 1 x 106 million Dalton's Lymphoma Ascites (DLA) tumour cells. At a 200 µg/ml concentration it registered 90% toxicity. Then its effect on the lifespan of mice with DLA tumour cells was studied. At high and low dosages of 50 and 10 mg. b. wt. kg-1 of herbal extract residue, it exhibited 87.5% and 60.4% increase in the lifespan, respectively. Blood parameters such as percentage Hb, RBC and WBC counts were conducted with tail vein blood samples. Hb and RBC counts of treated mice were higher than that of tumour bearing mice, while WBC counts were lower. This is a good index of tumour recovery. Further studies were carried out on mice with solid tumours to record volumes, along with a lifespan study. Low dosage of the herbal extract residue was able to control the tumour volume 35.1% and 32.9% increase in the lifespan was noted both at high and low dosages, respectively.

Photoinhibition of photosynthesis in needles of two cypress (Cupressus sempervirens) clones.

Photoinhibition of photosynthesis and photosynthetic recovery were studied in detached needles of cypress (Cupressus sempervirens L.) Clones 52 and 30 under controlled conditions of high irradiation (about 1900 micromol m(-2) s(-1) for 60 min; HL treatment), followed by 60 min in darkness. The degree of photoinhibition was determined based on the ratio of variable to maximum chlorophyll fluorescence (Fv/Fm), which is a measure of the potential efficiency of photosystem II (PSII), and on electron transport measurements. The Fv/Fm ratio declined in needles of both clones in response to the HL treatment. Minimal fluorescence (Fo) increased in HL-treated needles of both clones. The HL treatment decreased rates of whole-chain and PSII activity of isolated thylakoids more in Clone 52 than in Clone 30. In needles of both clones, PSI activity was less sensitive to photoinhibition than PSII activity. In the subsequent 60-min dark incubation, fast recovery was observed in needles of both clones, with PSII efficiencies reaching similar values to those in non-photoinhibited needles. The artificial exogenous electron donors diphenyl carbazide (DPC), hydroxylamine (NH2OH) and manganese chloride (MnCl2) failed to restore the HL-induced loss of PSII activity in needles of Clone 30, whereas DPC and NH2OH significantly restored PSII activity in photoinhibited needles of Clone 52. Quantification of the PSII reaction center protein D1 and the 33-kDa protein of the water-splitting complex following HL treatment of needles revealed pronounced differences between Clone 52 and Clone 30. The large decrease in PSII activity in HL-treated needles was caused by the marked loss of D1 protein and 33-kDa protein in Clone 30 and Clone 52, respectively.