Identification of unprecedented anticancer properties of high molecular weight biomacromolecular complex containing bovine lactoferrin (HMW-bLf)

With the successful clinical trials, multifunctional glycoprotein bovine lactoferrin is gaining attention as a safe nutraceutical and biologic drug targeting cancer, chronic-inflammatory, viral and microbial diseases. Interestingly, recent findings that human lactoferrin oligomerizes under simulated physiological conditions signify the possible role of oligomerization in the multifunctional activities of lactoferrin molecule during infections and in disease targeting signaling pathways. Here we report the purification and physicochemical characterization of high molecular weight biomacromolecular complex containing bovine lactoferrin (≥250 kDa), from bovine colostrum, a naturally enriched source of lactoferrin. It showed structural similarities to native monomeric iron free (Apo) lactoferrin (∼78-80 kDa), retained anti-bovine lactoferrin antibody specific binding and displayed potential receptor binding properties when tested for cellular internalization. It further displayed higher thermal stability and better resistance to gut enzyme digestion than native bLf monomer. High molecular weight bovine lactoferrin was functionally bioactive and inhibited significantly the cell proliferation (p<0.01) of human breast and colon carcinoma derived cells. It induced significantly higher cancer cell death (apoptosis) and cytotoxicity in a dose-dependent manner in cancer cells than the normal intestinal cells. Upon cellular internalization, it led to the up-regulation of caspase-3 expression and degradation of actin. In order to identify the cutting edge future potential of this bio-macromolecule in medicine over the monomer, its in-depth structural and functional properties need to be investigated further

Survivin Mutant Protects Differentiated Dopaminergic SK-N-SH Cells Against Oxidative Stress

Oxidative stress is due to an imbalance of antioxidant/pro-oxidant homeostasis and is associated with the progression of several neurological diseases, including Parkinson's and Alzheimer's disease and amyotrophic lateral sclerosis. Furthermore, oxidative stress is responsible for the neuronal loss and dysfunction associated with disease pathogenesis. Survivin is a member of the inhibitors of the apoptosis (IAP) family of proteins, but its neuroprotective effects have not been studied. Here, we demonstrate that SurR9-C84A, a survivin mutant, has neuroprotective effects against H2O2-induced neurotoxicity. Our results show that H2O2 toxicity is associated with an increase in cell death, mitochondrial membrane depolarisation, and the expression of cyclin D1 and caspases 9 and 3. In addition, pre-treatment with SurR9-C84A reduces cell death by decreasing both the level of mitochondrial depolarisation and the expression of cyclin D1 and caspases 9 and 3. We further show that SurR9-C84A increases the antioxidant activity of GSH-peroxidase and catalase, and effectively counteracts oxidant activity following exposure to H2O2. These results suggest for the first time that SurR9-C84A is a promising treatment to protect neuronal cells against H2O2-induced neurotoxicity

Chimeric aptamers in cancer cell-targeted drug delivery

Aptamers are single-stranded structured oligonucleotides (DNA or RNA) that can bind to a wide range of targets ("apatopes") with high affinity and specificity. These nucleic acid ligands, generated from pools of random-sequence by an in vitro selection process referred to as systematic evolution of ligands by exponential enrichment (SELEX), have now been identified as excellent tools for chemical biology, therapeutic delivery, diagnosis, research, and monitoring therapy in real-time imaging. Today, aptamers represent an interesting class of modern Pharmaceuticals which with their low immunogenic potential mimic extend many of the properties of monoclonal antibodies in diagnostics, research, and therapeutics. More recently, chimeric aptamer approach employing many different possible types of chimerization strategies has generated more stable and efficient chimeric aptamers with aptamer-aptamer, aptamer-nonaptamer biomacromolecules (siRNAs, proteins) and aptamer-nanoparticle chimeras. These chimeric aptamers when conjugated with various biomacromolecules like locked nucleic acid (LNA) to potentiate their stability, biodistribution, and targeting efficiency, have facilitated the accurate targeting in preclinical trials. We developed LNA-aptamer (anti-nucleolin and EpCAM) complexes which were loaded in iron-saturated bovine lactofeerin (Fe-blf)-coated dopamine modified surface of superparamagnetic iron oxide (Fe3O4) nanoparticles (SPIONs). This complex was used to deliver the specific aptamers in tumor cells in a co-culture model of normal and cancer cells. This review focuses on the chimeric aptamers, currently in development that are likely to find future practical applications in concert with other therapeutic molecules and modalities

Multidimensional Homeier's generalized class and its application to planar 1D Bratu problem

[EN] In this paper, a parametric family of iterative methods for solving nonlinear systems, including Homeier’s scheme is presented, proving its third-order of convergence. The numerical section is devoted to obtain an estimation of the solution of the classical Bratu problem by transforming it in a nonlinear system by using finite differences, and solving it with different elements of the iterative family.This research was supported by Ministerio de Economía y Competitividad MTM2014-52016-C02-02.Cordero Barbero, A.; Franqués García, AM.; Torregrosa Sánchez, JR. (2015). Multidimensional Homeier's generalized class and its application to planar 1D Bratu problem. Journal of the Spanish Society of Applied Mathematics. 70(1):1-10. https://doi.org/10.1007/s40324-015-0037-xS110701Abad, M. F., Cordero, A., Torregrosa, J. R.: Fourth-and fifth-order for solving nonlinear systems of equations: an application to the global positioning system, Abstr. Appl. Anal. (2013) (Article ID 586708)Andreu, C., Cambil, N., Cordero, A., Torregrosa, J.R.: Preliminary orbit determination of artificial satellites: a vectorial sixth-order approach, Abstr. Appl. Anal. (2013) (Article ID 960582)Awawdeh, F.: On new iterative method for solving systems of nonlinear equations. Numer. Algorithms 54, 395–409 (2010)Boyd, J.P.: One-point pseudospectral collocation for the one-dimensional Bratu equation. Appl. Math. Comput. 217, 5553–5565 (2011)Bratu, G.: Sur les equation integrals non-lineaires. Bull. Math. Soc. France 42, 113–142 (1914)Buckmire, R.: Applications of Mickens finite differences to several related boundary value problems. In: Mickens, R.E. (ed.) Advances in the Applications of Nonstandard Finite Difference Schemes, pp. 47–87. World Scientific Publishing, Singapore (2005)Cordero, A., Hueso, J.L., Martínez, E., Torregrosa, J.R.: A modified Newton-Jarratt’s composition. Numer. Algorithms 55, 87–99 (2010)Gelfand, I.M.: Some problems in the theory of quasi-linear equations. Trans. Am. Math. Soc. Ser. 2, 295–381 (1963)Homeier, H.H.H.: On Newton-tyoe methods with cubic convergence. J. Comput. Appl. Math. 176, 425–432 (2005)Jacobsen, J., Schmitt, K.: The Liouville-Bratu-Gelfand problem for radial operators. J. Differ. Equ. 184, 283–298 (2002)Jalilian, R.: Non-polynomial spline method for solving Bratu’s problem. Comput. Phys. Comm. 181, 1868–1872 (2010)Kanwar, V., Kumar, S., Behl, R.: Several new families of Jarratts method for solving systems of nonlinear equations. Appl. Appl. Math. 8(2), 701–716 (2013)Mohsen, A.: A simple solution of the Bratu problem. Comput. Math. with Appl. 67, 26–33 (2014)Petković, M., Neta, B., Petković, L., Džunić, J.: Multipoint Methods for Solving Nonlinear Equations. Academic Press, Amsterdam (2013)Sharma, J.R., Guna, R.K., Sharma, R.: An efficient fourth order weighted-Newton method for systems of nonlinear equations. Numer. Algorithms 62, 307–323 (2013)Sharma, J.R., Arora, H.: On efficient weighted-Newton methods for solving systems of nonlinear equations. Appl. Math. Comput. 222, 497–506 (2013)Traub, J.F.: Iterative Methods for the Solution of Equations. Chelsea Publishing Company, New York (1982)Wan, Y.Q., Guo, Q., Pan, N.: Thermo-electro-hydrodynamic model for electrospinning process. Int. J. Nonlinear Sci. Numer. Simul. 5, 5–8 (2004

Therapeutic Targeting of STAT3 (Signal Transducers and Activators of Transcription 3) Pathway Inhibits Experimental Autoimmune Uveitis

Mice with targeted deletion of STAT3 in CD4+ T-cells do not develop experimental autoimmune uveitis (EAU) or experimental autoimmune encephalomyelitis (EAE), in part, because they cannot generate pathogenic Th17 cells. In this study, we have used ORLL-NIH001, a small synthetic compound that inhibits transcriptional activity of STAT3, to ameliorate EAU, an animal model of human posterior uveitis. We show that by attenuating inflammatory properties of uveitogenic lymphocytes, ORLL-NIH001 inhibited the recruitment of inflammatory cells into the retina during EAU and prevented the massive destruction of the neuroretina caused by pro-inflammatory cytokines produced by the autoreactive lymphocytes. Decrease in disease severity observed in ORLL-NIH001-treated mice, correlated with the down-regulation of α4β1 and α4β7 integrin activation and marked reduction of CCR6 and CXCR3 expression, providing a mechanism by which ORLL-NIH001 mitigated EAU. Furthermore, we show that ORLL-NIH001 inhibited the expansion of human Th17 cells, underscoring its potential as a drug for the treatment of human uveitis. Two synthetic molecules that target the Th17 lineage transcription factors, RORγt and RORα, have recently been suggested as potential drugs for inhibiting Th17 development and treating CNS inflammatory diseases. However, inhibiting STAT3 pathways completely blocks Th17 development, as well as, prevents trafficking of inflammatory cells into CNS tissues, making STAT3 a more attractive therapeutic target. Thus, use of ORLL-NIH001 to target the STAT3 transcription factor, thereby antagonizing Th17 expansion and expression of proteins that mediate T cell chemotaxis, provides an attractive new therapeutic approach for treatment of posterior uveitis and other CNS autoimmune diseases mediated by Th17 cells

Maternal LAMP/p55gagHIV-1 DNA Immunization Induces In Utero Priming and a Long-Lasting Immune Response in Vaccinated Neonates

Infants born to HIV-infected mothers are at high risk of becoming infected during gestation or the breastfeeding period. A search is thus warranted for vaccine formulations that will prevent mother-to-child HIV transmission. The LAMP/gag DNA chimeric vaccine encodes the HIV-1 p55gag fused to the lysosome-associated membrane protein-1 (LAMP-1) and has been shown to enhance anti-Gag antibody (Ab) and cellular immune responses in adult and neonatal mice; such a vaccine represents a new concept in antigen presentation. In this study, we evaluated the effect of LAMP/gag DNA immunization on neonates either before conception or during pregnancy. LAMP/gag immunization of BALB/c mice before conception by the intradermal route led to the transfer of anti-Gag IgG1 Ab through the placenta and via breastfeeding. Furthermore, there were an increased percentage of CD4+CD25+Foxp3+T cells in the spleens of neonates. When offspring were immunized with LAMP/gag DNA, the anti-Gag Ab response and the Gag-specific IFN-γ-secreting cells were decreased. Inhibition of anti-Gag Ab production and cellular responses were not observed six months after immunization, indicating that maternal immunization did not interfere with the long-lasting memory response in offspring. Injection of purified IgG in conjunction with LAMP/gag DNA immunization decreased humoral and cytotoxic T-cell responses. LAMP/gag DNA immunization by intradermal injection prior to conception promoted the transfer of Ab, leading to a diminished response to Gag without interfering with the development of anti-Gag T- and B-cell memory. Finally, we assessed responses after one intravenous injection of LAMP/gag DNA during the last five days of pregnancy. The intravenous injection led to in utero immunization. In conclusion, DNA vaccine enconding LAMP-1 with Gag and other HIV-1 antigens should be considered in the development of a protective vaccine for the maternal/fetal and newborn periods