Wound healing activity of the hydroalcoholic extract and the main metabolites of Amphipterygium adstringens (cuachalalate) in a rat excision model.

ETHNOPHARMACOLOGICAL RELEVANCE: The bark of Amphipterygium adstringens Schiede ex Schltdl (Anacardiaceae), commonly known as 'cuachalalate' has been used in Mexican traditional medicine for the treatment of skin and oral lesions, gastric ulcers, and other conditions. The use as wound healing of the bark of this plant has been known since before the Spanish conquest of Mexico. Its uses are mentioned in the first writings of the Spanish in the 16th century. It is important to highlight that its use for wound healing treatment has no scientific previous reports. AIM OF THE STUDY: The objectives of this study were to determine the wound healing effect of the hydroalcoholic extract of the stem bark of Amphipterygium adstringens and its main metabolites, using a model of excision in the back of Wistar rats. To evaluate its antimicrobial effect against common bacteria that living on the skin of wounds and to evaluate its effect on angiogenesis. MATERIALS AND METHODS: The hydroalcoholic extract of cuachalalate (HAE, 10 mg/wound/day), the 3α-hydroxymasticadienoic acid (3 MA, 300 µg/wound/day), the masticadienoic acid (MA, 300 µg/wound/day), and a mixture of anacardic acids (ANA, 300 µg per wound) were tested in a murine excision model topically for 15 days, to evaluate their wound-healing effect. The results were reported in a wound closure percentage (n = 30 animals per treatment curve), using pirfenidone (PIR, 8% in vehicle) as a reference drug. In addition, histologic analysis was performed to evaluate the structure and quality of the scar. The effect on angiogenesis was assessed using the chick embryo chorioallantoic membrane (CAM) model (n = 6 eggs per treatment). The concentration evaluated for each treatment was 300 µg, using as proangiogenic reference drug the histamine (HIS, 5.6 µg) and as antiangiogenic drugs pirfenidone (9 µg) and acetylsalicylic acid (ASA, 9 µg). The antimicrobial test was performed against S. mutans, S. aureus, P. aeruginosa y E. coli using a minimum inhibitory concentration (MIC) assay. RESULTS: The 3α-hydroxymasticadienoic (3 MA) acid and the anacardic acids (ANA) improve the wound closure by approximates 30% (similar to pirfenidone) in comparison with the control-treated with the vehicle in the proliferative phase. On the other hand, the hydroalcoholic extract of cuachalalate (HAE) did not show an effect on the wound healing process. The histologic analysis demonstrated that the three main metabolites showed an improvement in the scar structure. According to the CAM results, it is probable that the main action mechanism of the 3α-hydroxymasticadienoic acid and the anacardic acids is related to their proangiogenic effect. In addition, ANA showed a modest antimicrobial effect. CONCLUSIONS: The 3α-hydroxymasticadienoic acid and anacardic acids showed a better tissue structure and reduced the time closure of the wound. In addition, the anacardic acids showed antimicrobial effects and both metabolites promote angiogenesis, suggesting that these effects may be related to their action mechanism. These metabolites of cuachalalate could be a good alternative for wound healing treatment.

Microenvironment-regulated lncRNA-HAL is able to promote stemness in breast cancer cells.

Multicellular Tumor Spheroids culture (MCTS) is an in vitro model mimicking the characteristics of the tumor microenvironment, such as hypoxia and acidosis, resulting in the presence of both proliferating and quiescent cell populations. lncRNA's is a novel group of regulatory molecules that participates in the acquisition of tumorigenic phenotypes. In the present work we evaluated the oncogenic association of an uncharacterized lncRNA (lncRNA-HAL) in the tumorigenic phenotype induced by the MCTS microenvironment. We measured lncRNA-HAL expression level in MCF-7-MCTS populations and under different hypoxic conditions by RT-qPCR. Afterwards, we silenced lncRNA-HAL expression by shRNAs and evaluated its effect in MCF-7 transcriptome (by RNAseq) and validated the modified cellular processes by proliferation, migration, and stem cells assays. Finally, we analyzed which proteins interacts with lncRNA-HAL by ChIRP assay, to propose a possible molecular mechanism for this lncRNA. We found that lncRNA-HAL is overexpressed in the internal quiescent populations (p27 positive populations) of MCF-7-MCTS, mainly in the quiescent stem cell population, being hypoxia one of the microenvironmental cues responsible of its overexpression. Transcriptome analysis of lncRNA-HAL knockdown MCF7 cells revealed that lncRNA-HAL effect is associated with proliferation, migration and cell survival mechanisms; moreover, lncRNA-HAL silencing increased cell proliferation and impaired cancer stem cell proportion and function, resulting in decreased tumor grafting in vivo. In addition, we found that this lncRNA was overexpressed in triple-negative breast cancer patients. Analysis by ChIRP assay showed that this nuclear lncRNA binds to histones and hnRNPs suggesting a participation at the chromatin level and transcriptional regulation. The results obtained in the present work suggest that the function of lncRNA-HAL is associated with quiescent stem cell populations, which in turn is relevant due to its implications in cancer cell survival and resistance against treatment in vivo. Altogether, our data highlights a new lncRNA whose expression is regulated by the tumor microenvironment and associated to stemness in breast cancer.

Promoting appropriate drug use through the application of the Spanish drug-related problem classification system in the primary care setting.

BACKGROUND: According to the Second Consensus of Granada (2002), the term drug-related problem (DRP) is defined as a health problem resulting from pharmacotherapy and is considered a negative clinical outcome (ie, a therapeutic objective is not achieved or adverse effects are reported). DRP classification systems used in primary care settings can be useful tools to detect, evaluate, and resolve DRPs. OBJECTIVE: To encourage appropriate drug use in the ambulatory clinical setting through DRP detection and evaluation by means of the Spanish DRP classification system, and to document how pharmacists can help resolve DRPs through interventions with both general practitioners (GPs) and patients. METHODS: Four pharmacists investigated DRPs in polymedicated patients over a 6-month period. All detected DRPs were grouped into 3 major categories: necessity, effectiveness, and safety. To resolve DRPs, pharmacists performed interventions on GPs and patients. GPs received verbal and written information about DRPs; patient interventions were in the form of private meetings on health education. RESULTS: Four hundred twenty-two patients, 80% of whom were aged 65 years or older, were included in the study. Each patient was taking a mean +/- SD of 8.1 +/- 2.4 medications. Three hundred four medications were associated with 245 DRPs; medications indicated for digestive/metabolic or cardiovascular pathologies were the most prevalent. Most (60%) of the identified DRPs belonged to the effectiveness category, whereas safety issues accounted for 28.6%. The most frequently reported DRP was pathology resistant to treatment (19.6%), followed by nonadherence (16.3%). Of the 215 interventions carried out to resolve these DRPs, 173 (80.5%) were addressed to GPs, who agreed to change therapy regimens on 90.2% of the occasions. Forty-two (19.5%) interventions were addressed to patients. Furthermore, the interventions accepted by GPs and patients resolved 176 (82%) DRPs. CONCLUSIONS: The current Spanish DRP classification system is a useful tool to systematically detect and document DRPs in daily general practice. In addition, the interventions addressed by pharmacists to GPs and patients resolved most of the detected DRPs.