Estimation of salivary proteins in early childhood caries before and after treatment using gel electrophoresis

Aerially Applied Zinc Oxide Nanoparticle Affects Reproductive Components and Seed Quality in Fully Grown Bean Plants ( Phaseolus vulgaris L.)

The development of reproductive components in plant species is susceptible to environmental stresses. The extensive application of zinc oxide nanoparticles (nZnO) in various agro-industrial processes has jeopardized the performance and functionality of plants. To understand Gentaur Gel Electrophoresis Systems the response of the developmental (gametogenesis and sporogenesis) processes to nanoparticles (NPs) exposure, the aerial application of nZnO and their ionic counterpart of ZnSO₄ at four different levels were examined on bean plants (Phaseolus vulgaris) before the flowering stage.

To evaluate the mentioned processes, briefly, flowers in multiple sizes were fixed in paraffin, followed by sectioning and optical analysis. The possibility of alteration in reproductive cells was thoroughly analyzed using both light and electron microscopes.

Overall, our results revealed the histological defects in male and female reproductive systems of mature plants depend on NPs levels. Furthermore, NPs caused tapetum abnormalities, aberrations in carbohydrate accumulation, and apoptosis. The nZnO induced abnormal alterations right after meiosis and partly hindered the microspore development, leading to infertile pollens. The seed yield and dry weight were reduced to 70 and 82% at 2,000 mg L^-1 nZnO foliar exposure, respectively. The sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis pattern showed the increased expression of two proteins at the molecular weight of 28 and 42 kDa at various concentrations of nZnO and ZnSO₄. Overall, our results provided novel insights into the negative effect of nano-scaled Zn on the differential mechanism involved in the reproductive stage of the plants compared with salt form.

A Guide to Analysis of Relative Synaptic Protein Abundance by Quantitative Fluorescent Western Blotting

The introduction of fluorescent detection systems has revolutionized the applicability of Western blotting for quantitative protein expression analyses. The fundamental premise behind fluorescent Western blotting is the combination of distinct fluorescent dye-conjugated secondary antibodies and high  performance digital imaging solutions in which the fluorescence signal is directly proportional to the amount of protein enabling quantitative measurements and simultaneous detection of several target proteins.

This aspect of Western blotting is now widely used, especially in preclinical research, to detect quantitative changes in protein levels and phosphorylation status between experimental groups. This chapter provides a detailed step-by-step guide for best practice procedures during the entire process from sample preparation, SDS polyacrylamide gel electrophoresis to electrotransfer of proteins and highlights approaches that can be applied to increase data output.

Overexpression of the dystrophins Dp40 and Dp40 _L170P modifies neurite outgrowth and the protein expression profile of PC12 cells

Dp40 is ubiquitously expressed including the central nervous system. In addition to being present in the nucleus, membrane, and cytoplasm, Dp40 is detected in neurites and postsynaptic spines in hippocampal neurons. Although Dp40 is expressed from the same promoter as Dp71, its role in the cognitive impairment present in Duchenne muscular dystrophy patients is still unknown.

Here, we studied the effects of overexpression of Dp40 and Dp40_L170P during the neuronal differentiation of PC12 Tet-On cells. We found that Dp40 overexpression increased the percentage of PC12 cells with neurites and neurite length, while Dp40_L170P overexpression decreased them compared to Dp40 overexpression.

Two-dimensional gel electrophoresis analysis showed that the protein expression profile was modified in nerve growth factor-differentiated PC12-Dp40_L170P cells compared to that of the control cells (PC12 Tet-On). The proteins α-internexin and S100a6, involved in cytoskeletal structure, were upregulated.

The expression of vesicle-associated membrane proteins increased in differentiated PC12-Dp40 cells, in contrast to PC12-Dp40_L170P cells, while neurofilament light-chain was decreased in both differentiated cells. These results suggest that Dp40 has an important role in the neuronal differentiation of PC12 cells through the regulation of proteins involved in neurofilaments and exocytosis of synaptic vesicles, functions that might be affected in PC12-Dp40_L170P.

Cytotoxic activity of Staphylococcus aureus isolates from a cohort of Mexican children with cystic fibrosis