Safety Assessment of Ethanolic Extract of Padina gymnospora as a Cosmetic Ingredient

Purpose: In a previous study, we identified the skin-whitening effect of the ethanolic extract of Padina gymnospora. The present study was performed to confirm the safety of the extract in animal replacement tests.Methods: To evaluate the safety of the extract of Padina gymnospora, the photosensitivity test (Harber test), in vitro 3T3 neutral red uptake (3T3 NRU) phototoxicity test, local lymph node assay (5-bromo-2'-deoxyuridine enzyme-linked immunosorbent assay), acute oral toxicity test, and reconstructed human epidermis (RHE) test were used. All experiments followed the guidelines of the Korean Ministry of Food and Drug Safety and were conducted by a GLP-certified organization (Chemon Inc.).Results: The extract of Padina gymnospora was not photosensitive: 0% photosensitization was detected (I grade: very weak). In the 3T3 NRU phototoxicity test, the relative viability of the extract-treated cells was higher than the guideline level; thus, the extract was classified as non-phototoxic. Treatment with the extract did not trigger skin irritation in the RHE test model and did not cause skin sensitization in the local lymph node assay. Finally, oral administration of the extract to rats indicated that it was not a harmful material as the LD50 was estimated at >2,000 mg/kg.Conclusion: The ethanolic extract of Padina gymnospora was demonstrated to be safe when applied to the skin. Taken together with our previous study of its efficacy, we conclude that this extract has the potential for use as a cosmetic ingredient.

Differentiation and modulation of phagocyte activity in murine granuloma after treatment with thymulin 5cH

In previous studies, it was found that thymulin (thymic hormone), when prepared in homeopathic 5CH dilution, can modulate diseases progress and immune processes, such as virus infection in poultry and experimental tumor development. This project aims to study the immune modulatory mechanisms of thymulin 5CH in granulomatous experimental model induced by BCG in mice, with special focus on the phagocytes differentiation from peritoneal B1 stem cells and on the activity of these cells and lymphocyte cooperation at the injury site. Male adult Balb/c mice were inoculated with BCG into the footpad to induce granuloma and its cells were evaluated by histomorphometric methods (Ziehl-Neelsen staining of macrophages) and flow cytometry. The phenotypic characterization of phagocytes, T and B lymphocytes in the peritoneum and regional lymph node was done. Thus, CD11b (activated phagocytes and B1 cells), CD19 (B1 cells and B2), CD23 (B2 cells), CD5 (B1 cell subtypes and T cells), CD4, CD8 (effective T lymphocytes) and CD25 (regulatory lymphocytes) positive cells were quantified. After 21 days of infection, the differentiation of B1 peritoneal stem cells into phagocytes reached the peak, being higher in thymulin-treated mice (X2, p=0.0001). Simultaneously, the number of infected phagocytes in the lesion decreased (KW, p=0.001), indicating remission of the infection. The number of B1 derived phagocytes, CD4+ and CD8+ T lymphocytes in the local lymph node increased in thymulin-treated mice (X2, p=0.0001), at the same time. No difference was seen regarding to CD25+ cells. The results show that thymulin 5CH treatment is able to improve the granuloma inflammatory process by modulating local and systemic phagocyte differentiation, as well as T cell migration into the local lymph node.

Evaluation of the Skin-Sensitizing Potential of Brazilian Green Propolis

Propolis is a resinous mixture produced by bees from their secretions and plant material, so its composition varies depending on its botanical origin. Propolis has several beneficial bioactivities, but its skin sensitization properties have long been suspected. Nevertheless, the skin sensitization potency of Brazilian green propolis (BGP) has not been scientifically evaluated. Here, we used scientifically reliable tests to evaluate it. In vitro antigenicity test based on the human cell line activation test (OECD TG 442E) was performed by measuring the expression of CD54 and CD86, which are indicators of the antigenicity of test substances, on THP-1 and DC2.4 cells. BGP did not affect the expression of either marker on THP-1 cells, but upregulated the expression of CD86 on DC2.4 cells, suggesting that BGP may be a skin sensitizer. Then, we performed local lymph node assay (LLNA, OECD TG 429) as a definitive in vivo test. LLNA showed that 1.70% BGP primed skin sensitization and is a “moderate sensitizer”. Our results indicate scientific proof of the validity of arbitrary concentrations (1–2%), which have been used empirically, and provide the first scientific information on the safe use of BGP.

BCAT1 Promotes Lung Adenocarcinoma Progression Through Enhanced Mitochondrial Respiration and NF-κB Pathway Activation

BCAT1 is up-regulated and acts as an oncogenic factor in many types of cancers, but its role in lung adenocarcinoma (LUAD) development is not clearly understood. Here we found BCAT1 protein level was up-regulated in tumor tissues, which was positively associated with TNM stage and local lymph node metastasis of LUAD patients. BCAT1 knockdown inhibited cell growth and mobility while BCAT1 overexpression promoted LUAD development both in vitro and in vivo. BCAAs metabolism and mitochondrial respiration were enhanced in BCAT1 overexpression cells, which were more sensitive to Leucine and Isoleucine supplements, compared to control cells. Moreover, RNA sequencing analysis suggested that differentially expressed genes (DEGs) in BCAT1 overexpression LUAD cells were enriched in metabolism, signal transduction, and immune response processes, and BCAT1 overexpression decreased NFKBIB mRNA level that induced NF-κB pathway activation in LUAD cells. As an inhibitor of NF-κB pathway, ammonium pyrrolidinedithiocarbamate (PDTC) treatment predominately counteracted the effect of NF-κB pathway activation and inhibited LUAD cells proliferation and migration, especially cells with BCAT1 overexpression. Taken together, our findings point a key role for BCAT1 in promoting LUAD development through metabolic reprogramming and NF-κB pathway activation, which provides promising molecular biomarker and therapeutic targets for LUAD diagnosis and treatment.

The downregulation of NCXs is positively correlated with the prognosis of stage II–IV colon cancer

Purpose Colon cancer (CC) is a very common gastrointestinal tumor that is prone to invasion and metastasis in the late stage. This study aims to observe the expression of Na+/Ca2+ exchangers (NCXs) and analyze the correlation between NCXs and the prognosis of CC. Methods Specimens of 111 stage II–IV CC patients were collected. We used western blotting, qPCR, and immunohistochemical staining to observe the distributions and expression levels of NCX isoforms (NCX1, NCX2, and NCX3) in CC and distal normal tissues. Cox proportional hazards models were used to assess prognostic factors for patients. Results The expression of NCXs in most tumor specimens was lower than that in normal tissues. The NCX expression levels in tumor tissues from the primary tumor, local lymph node metastasis sites, and distant liver metastasis sites were increasingly significantly lower than those in normal tissues. The results of the Kaplan-Meier survival curves showed that the downregulation of any NCX isoform was closely related to the worse prognosis of advanced CC. Conclusion NCXs can be used as independent prognostic factors for CC. Our research results are expected to provide new targets for the treatment of CC.

Development of a PET/CT Molecular Radiomics-clinical Model to Predict Local Lymph Node Metastasis of Invasive Lung Adenocarcinoma (≤ 3cm)

Purpose To investigate the value of 18F-FDG PET/CT molecular radiomics combined with the clinical model in predicting local lymph node metastasis (LLNM) with invasive lung adenocarcinoma (≤3cm). Methods 528 lung adenocarcinoma patients were enrolled in this retrospective study. Five models, including integrated PET/CT molecular radiomics-clinical, PET/CT radiomics, PET radiomics, CT radiomics, and clinical models, were developed for the prediction of LLNM. The predictive performance was examined by ROC curve analysis and clinical utility was validated by nomogram and decision curve analysis (DCA) analyses. Results 10 PET/CT radiomics features and 2 clinical characteristics were selected for the construction of the integrated PET/CT molecular radiomics-clinical model. This integrated model performed better than the clinical model and three other radiomics models, and the AUC value of the integrated model was 0.95 (95% CI: 0.93-0.97) in the training group and 0.94 (95% CI: 0.89-0.97) in the test group, respectively. The clinical application of this integrated model in predicting LLNM was also confirmed by nomogram and DCA analyses. Conclusions The integrated PET/CT molecular radiomic-clinical model developed here has the greater advantage to predict LLNM of clinical invasive lung adenocarcinoma (≤3cm) when compared with the simple radiomics model or clinical model.

Quantification of Brucella abortus population structure in a natural host

Cattle are natural hosts of the intracellular pathogen Brucella abortus, which inflicts a significant burden on the health and reproduction of these important livestock. The primary routes of infection in field settings have been described, but it is not known how the bovine host shapes the structure of B. abortus populations during infection. We utilized a library of uniquely barcoded B. abortus strains to temporally and spatially quantify population structure during colonization of cattle through a natural route of infection. Introducing 108 bacteria from this barcoded library to the conjunctival mucosa resulted in expected levels of local lymph node colonization at a 1-wk time point. We leveraged variance in strain abundance in the library to demonstrate that only 1 in 10,000 brucellae introduced at the site of infection reached a parotid lymph node. Thus, cattle restrict the overwhelming majority of B. abortus introduced via the ocular conjunctiva at this dose. Individual strains were spatially restricted within the host tissue, and the total B. abortus census was dominated by a small number of distinct strains in each lymph node. These results define a bottleneck that B. abortus must traverse to colonize local lymph nodes from the conjunctival mucosa. The data further support a model in which a small number of spatially isolated granulomas founded by unique strains are present at 1 wk postinfection. These experiments demonstrate the power of barcoded transposon tools to quantify infection bottlenecks and to define pathogen population structure in host tissues.