Glycosylation differences of an anti-VEGF monoclonal antibody (PRO-169) and its extensive comparison with Bevacizumab.

PRO-169 is an anti-VEGF monoclonal antibody developed by Laboratorios Sophia that shares its sequence with Bevacizumab (BVZ); though, PRO-169 is intended for intravitreal administration. In this study, analytical characterization showed that PRO-169 had glycosylation differences in comparison to BVZ reference product (RP); since it had more content of G1F, G2F, sialic acid and high mannose. Further investigation was performed to evaluate if differences between both products would affect the efficacy and safety profile of PRO-169. PRO-169 had no alteration in its in vitro biological activity; moreover, no cytotoxicity or immunogenicity concerns should be expected as demonstrated by different orthogonal methods at analytical, in vitro and in vivo assays. These results support moving to the clinical testing of PRO-169 since no major complications will be expected with its clinical use for the treatment of ophthalmic diseases.

Physicochemical characterization of a DMPC-based nanoemulsion for dry eye and compatibility test with soft contact lenses in vitro.

INTRODUCTION: The use of contact lenses (CL) is often associated with hallmark symptoms of dry eye disease (DED) such as red eye and dryness. Even though lipid-based artificial tears are already marketed for DED, there is little evidence that supports their use while wearing soft CL. METHODS: An oil in water (O/W) nanoemulsion was formulated with a highly-stable oily phase composed of castor oil and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). Physicochemical characterization of the DMPC-based nanoemulsion (DMPC-NE) was performed using the Zetasizer Nano ZSP, and its long-term stability was evaluated over 24 months; in addition, the in vitro cytotoxicity of DMPC-NE was determined by Neutral Red Uptake (NRU) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Finally, the physical compatibility of the DMPC-NE with soft CL was tested by measuring the power, diameter, thickness and UV transmittance of two different types of CL. RESULTS: DMPC-NE had a mean particle size of 86.48 ± 4.22 nm, a polydispersity index of 0.22 ± 0.01 and a zeta potential of -33.23 ± 0.93 mV that remained with no changes after 24 months. DMPC-NE formulation, free of preservatives, showed no cytotoxicity and it was compatible with the physical properties of FDA-class II and -class IV CL. CONCLUSION: DMPC-NE is a highly stable formulation for dry eye that is safe to be used while wearing soft CL.

ß-Caryophyllene Reduces DNA Oxidation and the Overexpression of Glial Fibrillary Acidic Protein in the Prefrontal Cortex and Hippocampus of d-Galactose-Induced Aged BALB/c Mice.

Aging is associated with detrimental cellular and cognitive changes, making it an important public health concern; yet, many of these changes may be influenced by nutritional interventions. The natural sesquiterpene ß-caryophyllene (BCP) has anti-inflammatory and antioxidant effects that are mediated by cannabinoid type-2 receptor activation, and these actions promote neuroprotection in different animal models that involve a cognitive damage. Consequently, whether chronic administration of BCP might prevent the age-related cellular and cognitive damage in a model of aging induced by chronic d-galactose (GAL) consumption was assessed here. Male BALB/c mice were administered BCP (10 mg/kg, oral), GAL (300 mg/kg, intraperitoneal), or GAL+BCP, and long-term memory and cognitive flexibility were evaluated in the normal and the reverse phases of Morris water maze test. In addition, immunohistochemistry was performed on prefrontal and hippocampal brain slices to detect glial acidic fibrillary protein and DNA oxidation. Although GAL administration reduced cognitive flexibility (P = .0308), this functional damage was not reversed by administering BCP. However, GAL administration also elevated the total number of astrocytes and their interactions in the hippocampus, and increasing DNA oxidation in the prefrontal cortex. BCP administration impeded the rise in the total number of astrocytes (P = .0286) and the DNA oxidation (P = .0286) in mice that received GAL. Hence, although BCP did not improve cognitive flexibility, it did produce a neuroprotective effect at the molecular and cellular level in the GAL model of aging.

Ascorbic acid, ultraviolet C rays, and glucose but not hyperthermia are elicitors of human ß-defensin 1 mRNA in normal keratinocytes.

Hosts' innate defense systems are upregulated by antimicrobial peptide elicitors (APEs). Our aim was to investigate the effects of hyperthermia, ultraviolet A rays (UVA), and ultraviolet C rays (UVC) as well as glucose and ascorbic acid (AA) on the regulation of human ß-defensin 1 (DEFB1), cathelicidin (CAMP), and interferon-γ (IFNG) genes in normal human keratinocytes (NHK). The indirect in vitro antimicrobial activity against Staphylococcus aureus and Listeria monocytogenes of these potential APEs was tested. We found that AA is a more potent APE for DEFB1 than glucose in NHK. Glucose but not AA is an APE for CAMP. Mild hypo- (35°C) and hyperthermia (39°C) are not APEs in NHK. AA-dependent DEFB1 upregulation below 20 mM predicts in vitro antimicrobial activity as well as glucose- and AA-dependent CAMP and IFNG upregulation. UVC upregulates CAMP and DEFB1 genes but UVA only upregulates the DEFB1 gene. UVC is a previously unrecognized APE in human cells. Our results suggest that glucose upregulates CAMP in an IFN-γ-independent manner. AA is an elicitor of innate immunity that will challenge the current concept of late activation of adaptive immunity of this vitamin. These results could be useful in designing new potential drugs and devices to combat skin infections.