Dr. Hepp-Castro, Matías

Radiata pine bark is a widely available organic waste, requiring alternative uses due to its environmental impact on soil, fauna, and forest fires. Pine bark waxes could be used as cosmetic substitutes, but their toxicity requires evaluation since pine bark may contain toxic substances or xenobiotics, depending on the extraction process. This study evaluates the toxicity of radiata pine bark waxes obtained through various extraction methods on human skin cells grown in vitro. The assessment includes using XTT to evaluate mitochondrial activity, violet crystal dye to assess cell membrane integrity, and ApoTox-Glo triple assay to measure cytotoxicity, viability, and apoptosis signals. Pine bark waxes extracted via T3 (acid hydrolysis and petroleum ether incubation) and T9 (saturated steam cycle, alkaline hydrolysis, and petroleum ether incubation) exhibit non-toxicity up to 2% concentration, making them a potential substitute for petroleum-based cosmetic materials. Integrating the forestry and cosmetic industries through pine bark wax production under circular economy principles could promote development while replacing petroleum-based materials. Extraction methodology affects pine bark wax toxicity in human skin cells due to the retention of xenobiotic compounds including methyl 4-ketohex-5-enoate; 1-naphthalenol; dioctyl adipate; eicosanebioic acid dimethyl ester; among others. Future research will investigate whether the extraction methodology alters the molecular structure of the bark, affecting the release of toxic compounds in the wax mixture.

The bark of Pinus radiata D. Don is a bioresource of great worldwide abundance. While various forms of use have been studied, it is still a little-used bioresource. Due to its great accumulation, significant solid emissions of this residue generate environmental problems such as changes in soil chemistry, ecological problems such as the alteration of arthropod communities and fire risk. The opportunity to take advantage of this bioresource could be in its wax content, which could be a replacement for the main raw materials used in the production of cosmetics. These currently correspond to petroleum-derived substances, such as petroleum jelly, paraffin, or mineral oil. The importance of replacing these raw materials is that several studies report that they are the main causes of human skin diseases, such as chemical hypersensitivity syndrome and allergic contact dermatitis. This study seeks to prove that lipophilic extracts (waxes) from pine bark can replace petroleum-derived raw materials used in cosmetics. To achieve this, pine bark at drying conditions was characterized, and the performance of wax extraction by various treatments was studied. The density, viscosity, melting point, and solubility in culture media of the obtained waxes were determined. The waxes were chemically characterized by FT-IR and GC–MS analysis. The results reveal that the maximum moisture of the bark is 14.54 %, the best extraction yields are obtained by using water at 120 ◦C and 1.2 atm, and petroleum ether (3.12 %), alkaline hydrolysis 1 mol L− 1 (NaOH) and petroleum ether (3.53 %) ethyl acetate (3.23 %). Values were close to the reference study using the rapid lipid extraction method. The density of the wax is 0.845 g mL− 1 and its viscosity of 530 cP (24 ◦C), and the melting point varied according to the extractive treatment at between 25 and 40 ◦C. Solubility tests made it possible to determine that the 10/50/1000 μL ratio of modified Eagle Dulbecco wax/dimethylsulfoxide/medium allows homogeneous solubilization of the wax without the presence of precipitates. Chemical characterization identified typical functional groups of plant-based waxes such as long-chain alkanes, alkyls, methyl groups, esters, and carbonyls, with the most abundant fatty acids being C:22 and C:24.

Colorectal cancer (CRC) is one of the most common malignant neoplasms and the second leading cause of death from tumors worldwide. Therefore, there is a great need to study new therapeutical strategies, such as effective immunotherapies against these malignancies. Unfortunately, many CRC patients do not respond to current standard immunotherapies, making it necessary to search for adjuvant treatments. Histone deacetylase 6 (HDAC6) is involved in several processes, including immune response and tumor progression. Specifically, it has been observed that HDAC6 is required to activate the Signal Transducer and Activator of Transcription 3 (STAT3), a transcription factor involved in immunogenicity, by activating different genes in these pathways, such as PD-L1. Over-expression of immunosuppressive pathways in cancer cells deregulates T-cell activation. Therefore, we focused on the pharmacological inhibition of HDAC6 in CRC cells because of its potential as an adjuvant to avoid immunotolerance in immunotherapy. We investigated whether HDAC6 inhibitors (HDAC6is), such as Nexturastat A (NextA), affected STAT3 activation in CRC cells. First, we found that NextA is less cytotoxic than the non-selective HDACis panobinostat. Then, NextA modified STAT3 and decreased the mRNA and protein expression levels of PD-L1. Importantly, transcriptomic analysis showed that NextA treatment affected the expression of critical genes involved in immunomodulatory pathways in CRC malignancies. These results suggest that treatments with NextA reduce the functionality of STAT3 in CRC cells, impacting the expression of immunomodulatory genes involved in the inflammatory and immune responses. Therefore, targeting HDAC6 may represent an interesting adjuvant strategy in combination with immunotherapy.

Diverse factors play roles in chromatin dynamics, including linker proteins. Among them are high mobility group (HMG) box family proteins and linker histones. In the yeast Saccharomyces cerevisiae, Hmo1 has been identified as an HMG-box protein. This protein displays properties that are in agreement with this allocation. However, a number of studies have postulated that Hmo1 functions as a linker histone in yeast. On the other hand, when discovered, the Hho1 protein was identified as a linker histone. While multiple studies support this classification, some findings point to characteristics of Hho1 that are dissimilar to those commonly assigned to linker histones. In order to better understand the roles played by Hmo1 and Hho1 in chromatin dynamics and transcriptional regulation, we performed several analyses directly comparing these two proteins. Our analyses of genome-wide binding profiles support the belonging of Hmo1 to the HMGB family and Hho1 to the linker histones family. Interestingly, by performing protein-protein interaction analyses we found that both Hmo1 and Hho1 display physical interaction with the ATP-dependent chromatin remodeling complexes RSC, ISW1a and SWI/SNF. Moreover, by carrying out nucleosome remodeling assays, we found that both proteins stimulate the activity of the ISW1a complex. However, in the case of RSC, Hmo1 and Hho1 displayed differential properties, with Hho1 mainly showing an inhibitory effect. Our results are in agreement with the opposite roles played by RSC and ISW1a in chromatin dynamics and transcriptional regulation, and expand the view for the roles played by Hho1 and linker histones.