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Background: Interferons (IFNs) are cytokines involved in the immune response with a synergistic regulatory effect on the immune response. They are therapeutics for various viral and proliferative conditions, with proven safety and efficacy. Their clinical application is challenging due to the molecules' size, degradation, and pharmacokinetics. We are working on new drug delivery systems that provide adequate therapeutic concentrations for these cytokines and prolong their half-life in the circulation, such as nanoformulations. Methods: Through nanoencapsulation using electrospray technology and biocompatible and biodegradable polymers, we are developing a controlled release system based on nanoparticles for viral infections of the respiratory tract. Results: We developed a controlled release system for viral respiratory tract infections. A prototype nanoparticle with a core was created, which hydrolyzed the polyvinylpyrrolidone (PVP) shell , releasing the active ingredients interferon-alpha (IFN-α) and interferon-gamma (IFN-γ). The chitosan (QS) core degraded slowly, with a controlled release of IFN-α. The primary and rapid effect of the interferon combination ensured an antiviral and immunoregulatory response from day one, induced by IFN-α and enhanced by IFN-γ. The multilayer design demonstrated an optimal toxicity profile. Conclusions: This formulation is an inhaled dry powder intended for the non-invasive intranasal route. The product does not require a cold chain and has the potential for self-administration in the face of emerging viral infections. This novel drug has applications in multiple infectious, oncological, and autoimmune conditions, and further development is proposed for its therapeutic potential. This prototype would ensure greater bioavailability, controlled release, fewer adverse effects, and robust biological action through the simultaneous action of both molecules. [ABSTRACT FROM AUTHOR]

Pig is one of the most consumed meats worldwide. One of the main conditions for pig production is Porcine Enteropathy caused by Lawsonia intracellularis. Among the effects of this disease is chronic mild diarrhea, which affects the weight gain of pigs, generating economic losses. Vaccines available to prevent this condition do not have the desired effect, but this limitation can be overcome using adjuvants. Pro-inflammatory cytokines, such as interleukin 18 (IL-18), can improve an immune response, reducing the immune window of protection. In this study, recombinant porcine IL-18 was produced and expressed in Escherichia coli and Pichia pastoris. The protein's biological activity was assessed in vitro and in vivo, and we determined that the P. pastoris protein had better immunostimulatory activity. A vaccine candidate against L. intracellularis, formulated with and without IL-18, was used to determine the pigs' cellular and humoral immune responses. Animals injected with the candidate vaccine co-formulated with IL-18 showed a significant increase of Th1 immune response markers and an earlier increase of antibodies than those vaccinated without the cytokine. This suggests that IL-18 acts as an immunostimulant and vaccine adjuvant to boost the immune response against the antigens, reducing the therapeutic window of recombinant protein-based vaccines. [ABSTRACT FROM AUTHOR]

The most important mechanisms in the blood pressure regulation, fluid volume and sodium-potassium balance in humans is the renin-angiotensin-aldosterone system (RAAS). This regulatory pathway plays a critical role in modulating cardiac function and vascular tone. An alteration in any of the molecules that make up this system (RAAS) could contribute to the development of arterial hypertension. In this important mechanisms, the neutral endopeptidase, also called Neprilysin (NEP) is the main enzyme for the degradation of natriuretic, therefore, it is essential proteins in controlling blood pressure. In this work, we have used docking methodology, molecular dynamics simulationa and free energy calculations method (MM-PBSA), to comprehensively evaluate the inhibitory behavior of some ligands obteined from consulted literature. The principal results obtained shown these ligands were adequately oriented in the NEP pocket. The Lig783, Lig2177, and Lig3444 compounds were those with better dynamic behavior. The energetic components that contribute to the complex's stability are the electrostatic and Van der Waals components; however, when the ADME-Tox properties were analyzed, we conclude that the best possible anti-hypertensive candidate are Lig783 and Lig3444. [ABSTRACT FROM AUTHOR]

P ichia pastoris is a highly successful system for the large-scale expression of heterologous proteins, with the added capability of performing most eukaryotic post-translational modifications. However, this system has one significant disadvantage - frequent proteolytic degradation by P . pastoris proteases of heterologously expressed proteins. Several methods have been proposed to address this problem, but none has proven fully effective. We tested the effectiveness of a broad specificity protease inhibitor to control proteolysis. A recombinant variant of the BPTI- Kunitz protease inhibitor Sh PI-1 isolated from the sea anemone S tichodactyla helianthus, was expressed in P . pastoris. The recombinant inhibitor (r Sh PI-1 A), containing four additional amino acids ( EAEA) at the N-terminus, was folded similarly to the natural inhibitor, as assessed by circular dichroism. r Sh PI-1 A had broad protease specificity, inhibiting serine, aspartic, and cysteine proteases similarly to the natural inhibitor. r Sh PI-1 A protected a model protein, recombinant human miniproinsulin (rh MPI), from proteolytic degradation during expression in P . pastoris. The addition of purified r Sh PI-1 A at the beginning of the induction phase significantly protected rh MPI from proteolysis in culture broth. The results suggest that a broad specificity protease inhibitor such as r Sh PI-1 A can be used to improve the yield of recombinant proteins secreted from P . pastoris. [ABSTRACT FROM AUTHOR]