Macrocyclic amphiphilic molecules based on calix[4]arenes are highly attractive for controlled supramolecular assembly of DNA into small nanoparticles, since they present a unique conical architecture and can bear multiple charged groups. In the present work, we synthesized new amphiphilic calixarenes bearing cationic groups at the upper rim and alkyl chains at the lower rim. Their self-assembly in aqueous solution was characterized by fluorescent probes, fluorescence correlation spectroscopy, dynamic light scattering, gel electrophoresis and atomic force microscopy. We found that calixarenes bearing long alkyl chains (octyl) self-assemble into micelles of 6 nm diameter at low critical micellar concentration and present the unique ability to condense DNA into small nanoparticles of about 50 nm diameter. In contrast, the short-chain (propyl) analogues that cannot form micelles at low concentrations failed to condense DNA, giving large polydisperse DNA complexes. Thus, formation of small DNA nanoparticles is hierarchical, requiring assembly of calixarenes into micellar building blocks that further co-assemble with DNA into small virus-sized particles. The latter showed much better gene transfection efficiency in cell cultures relative to the large DNA complexes with the short-chain analogues, which indicates that gene delivery of calixarene/DNA complexes depends strongly on their structure. Moreover, all cationic calixarenes studied showed low cytotoxicity. Thus, this work presents a two-step hierarchical assembly of small DNA nanoparticles for gene delivery based on amphiphilic cone-shaped cationic calixarenes.

This study was conducted to explore whether hyperspectral data could be used to discriminate between the effects of different rates of nitrogen application to a potato crop. The field experiment was carried out in the Central Potato Research Station, Jalandhar, on seven plots with different nitrogen (N) treatments. Spectral reflectance was measured using a 512-channel spectroradiometer with a range of 395-1075 nm on two different dates during crop growth. An optimum number of bands were selected from this range based on band-band r 2 , principal component analysis and discriminant analysis. The four bands that could discriminate between the rates of N applied were 560, 650, 730, and 760 nm. An ANOVA analysis of several narrow-band indices calculated from the reflectance values showed the indices that were able to differentiate best between the different rates of N application. These were reflectance ratio at the red edge (R 740/720 ) and the structure insensitive pigment index (SIPI). To estimate leaf N, reflectance ratios were determined for each band combination and were evaluated for their correlation with the leaf N content. A regression model for N estimation was obtained using the reflectance ratio indices at 750 and 710 nm wavelengths (F-ratio = 32 and r 2 = 0.551, P \ 0.000).

The study was carried out to investigate the utility of hyperspectral reflectance data for potato late blight disease detection. The hyperspectral data was collected for potato crop at different level of disease infestation using hand-held spectroradiometer over the spectral range of 325-1075 nm. The data was averaged into 10-nm wide wavebands, resulting in 75 narrowbands. The reflectance curve was partitioned into five regions, viz. 400-500 nm, 520-590 nm, 620-680 nm, 770-860 nm and 920-1050 nm. The notable differences in healthy and diseased potato plants were noticed in 770-860 nm and 920-1050 nm range. Vegetation indices, namely NDVI, SR, SAVI and red edge were calculated using reflectance values. The differences between the vegetation indices for plants at different levels of disease infestation were found highly significant. The optimal hyperspectral wavebands to discriminate the healthy plants from disease infested plants were 540, 610, 620, 700, 710, 730, 780 and 1040 nm whereas upto 25% infestation could be discriminated using reflectance at 710, 720 and 750 nm.

The success in gene therapy relies strongly on new efficient gene delivery vectors. Nonviral vectors based on lipids and polymers constitute an important alternative to the viral vectors. However, the key problem with these vectors is the poor structural control of their DNA complexes. In the present work, following new design we synthesized unsymmetrical bolaamphiphiles, molecules bearing neutral sugar (gluconic acid) and dicationic ornithine head groups connected by different long hydrophobic spacers. Within this design, a positively charged headgroup is expected to bind DNA, the hydrophobic spacer is to drive the formation of a monolayer membrane shell around DNA, while the neutral group is to be exposed outside of the complex. Our fluorescence and gel electrophoresis data showed that self-assembly of bolas and their interaction with DNA depend strongly on the bola structure. The size of bola/DNA complexes (bolaplexes) estimated from dynamic light scattering data was ∼100 nm at low N/P (cationic nitrogen/DNA phosphate molar ratio), while at higher N/Ps it was significantly larger due to neutralization of their surface charge. Atomic force microscopy studies revealed nanostructural rod-shaped or spherical morphology of the bolaplexes. Transfection efficiency of the bolaplexes in vitro was significant when either DOPE or chloroquine were used as helping agents, suggesting that the key barrier for their internalization is the endosomal escape. Finally, all bolas showed low cytotoxicity (cell viability >80%). The present results show that bolas are prospective candidates for construction of nonviral gene delivery vectors. We believe that further optimization of polar head groups and a hydrophobic spacer in the bolas will lead to vectors with controlled small size and high transfection efficiency.