The Value of CXCL1, CXCL2, CXCL3, and CXCL8 as Potential Prognosis Markers in Cervical Cancer: Evidence of E6/E7 from HPV16 and 18 in Chemokines Regulation.

Cervical cancer (CC) is a serious global health issue, and it is well-known that HPV infection is the main etiological factor that triggers carcinogenesis. In cancer, chemokine ligands and receptors are involved in tumor cell growth, metastasis, leukocyte infiltration, and angiogenesis; however, information on the role played by E6/E7 of HPV16/18 in the modulation of chemokines is very limited. Therefore, this study aimed to determine whether chemokines are differentially expressed in CC-derived cell lines; if E6/E7 oncoproteins from HPV16 and 18 are capable of mediating chemokine expression, what is the expression profile of chemokines in tissues derived from CC and what is their impact on the overall survival of patients with this pathology? For this purpose, RNA sequencing and real-time PCR were performed on SiHa, HeLa, and C33A tumorigenic cell lines, on the non-tumorigenic HaCaT cells, and the E6/E7 HPV-transduced HaCaT cell models. Furthermore, chemokine expression and survival analysis were executed on 304 CC and 22 normal tissue samples from The Cancer Genome Atlas (TCGA) repository. The results demonstrate that CXCL1, CXCL2, CXCL3, and CXCL8 are regulated by E6/E7 of HPV16 and 18, are overexpressed in CC biopsies, and that their higher expression is related to a worse prognostic survival.

E6/E7 from Beta-2-HPVs 122, 38b, and 107 possess transforming properties in a fibroblast model in vitro.

Beta-2 Human papillomaviruses 38b, 107, and 122 have been frequently found in cervical cancer samples in western Mexico. Because their E6/E7 genes functions are not fully elucidated, we deepen into their transformation capabilities. To achieve this goal, primary human fibroblasts (FB) were transduced with E6/E7 genotype-specific viral particles. Additionally, E6/E7 from HPVs 16 and 18 were included as controls. All E6/E7-cell models increased their lifespan; however, it is important to highlight that FB-E6/E7-122 showed growth as accelerated as FB-E6/E7-16 and 18. Furthermore, both FB-E6/E7-38b and 122 exhibited abilities to migrate, and FB-E6/E7-122 presented high invasive capacity. On the other hand, ΔNp73 expression was found in all cell models, except for FB-pLVX (empty-vector). Finally, RNAseq found differentially expressed genes enriched in signaling pathways related to cell cycle, epithelial-mesenchymal transition, and cancer, among others. This study shows for the first time, the great transformative potential that genotypes of the Beta-2 also possess, especially HPV122. These Beta-2 HPVs can modulate some of the genes that are well known to be regulated by Alpha-HPVs, however, they also possess alternative strategies to modulate diverse signaling pathways. These data support the idea that Beta-2 HPVs should play an important role in co-infections with Alpha-HPV during carcinogenesis.

Towards a model of wet deposition of bioaerosols: The raindrop size role.

Bioaerosols play a major role in the plant life of ecosystems. In addition, they have a profound impact on human health, since they may cause lung diseases or allergies. The key objective of this study is to assess the below cloud scavenging effect of rainfall on pollen concentration. The analysis is based on a sampling carried out in León, Spain, between 2015 and 2018. The rainfall variables and the pollen concentrations have been obtained with a disdrometer and a volumetric Hirst type spore-trap, respectively. In order to evaluate the scavenging, three parameters have been calculated: scavenging efficiency (through the concentration-weighted average (%ΔC)), the scavenging coefficient (λ) and the percentage of events with a decrease in pollen concentration (%ES) also called events with effective scavenging. 71% of rain events presented an effective scavenging that affected all types of pollen. The %ΔC mean value of total pollen was 24 ± 18% (positive values indicate an effective scavenging) and the types of pollen with the highest values were Castanea and Cupressaceae (71 and 40%, respectively). A linear model (R2 = 0.94) to estimate the pollen concentration after rain was built with variables such as pollen concentration before rain and other variables from a weather station and a disdrometer. Furthermore, we have shown the possibility of knowing in real time the probable Cupressaceae pollen concentration, from the initial pollen concentration and the physical parameters of rain (such as raindrop size, rain intensity or volume swept by raindrops in their falling path).

Towards a model for aerosol removal by rain scavenging: The role of physical-chemical characteristics of raindrops.

A one-year study was carried out in León, Spain, in order to characterize physically and chemically the precipitation. With the aim of studying the scavenging process of atmospheric pollutants, scavenging ratio and removal coefficients were calculated through physical parameters of raindrops (obtained by disdrometer data) and through chemical properties of aerosols. Finally, linear models for the prediction of the chemical composition of rainwater and the efficiency of the removal effect were established. In general, the rainwater was dominated by NH4+ > SO42- > NO3- in all seasons. Higher ion concentrations and conductivity and lowest pH were observed in summer, due to the low volume of rain. In winter, the high values of Na+ and Cl- in the rainwater showed the contribution from marine sources, while in summer the high concentrations of Ca2+, Mg2+, SO42-, NH4+ and NO3- reflected the contribution from both crustal and anthropogenic sources. The linear models revealed that the amount of dissolved organic carbon and of the water-soluble ions in rain samples, Ca2+, SO42-, NO3-, increases with the volume swept by the falling drops. Insoluble carbon fraction has a negative dependence with the volume swept and positive with the diameter of the raindrop. Removal coefficients are affected by the concentration in the air of each species before precipitation, the duration of the event and the time elapsed between two precipitation events.

Characterization of aerosol sources in León (Spain) using Positive Matrix Factorization and weather types.

A one-year aerosol sampling campaign, between 2016 and 2017, was conducted in a suburban area of León city, Spain. An association between the Positive Matrix Factorization (PMF) results and air masses through circulation weather types was carried out, through the construction of linear models from the PM10 concentrations and its chemical composition. The aerosol sources, identified by PMF six-factor solution, were: traffic (29%), aged sea salt (26%), secondary aerosols (16%), dust (13%), marine aerosol (7%) and biomass burning (3%). Traffic and secondary factors showed the highest PM10 contribution in the hybrid cyclonic types with wind component from the first and second quadrant. Anticyclonic types with wind component from the first quadrant exhibited high values of secondary, aged sea salt and dust factors. The highest contributions of the dust factor were also associated with northerly types. The linear models built for estimating the source apportionment of PM10, from aerosol chemical composition and geostrophic flow, showed positive coefficients for: westerly flows (WF) in marine factor, southerly flows (SF) in secondary and traffic factors, and shear southerly vorticities (ZS) in dust factor. Negative dependences were observed for ZS in aged sea salt factor and for SF in dust factor. The PM10 mass concentration calculated by the linear models and by the PMF model were strongly correlated. This can be very useful to determine the contribution of a specific source to PM10 in León, only by knowing some meteorological and chemical variables.

Partial Fat Replacement by Boiled Quinoa on the Quality Characteristics of a Dry-Cured Sausage.

Different approaches have been previously studied in order to reduce the fat content of dry-cured sausages. Among them, the use of polysaccharides, such as fiber, gums, or starch, have been proposed for fat replacing. Although scarcely studied, it is likely that starchy grains and vegetables might also be used as potential fat replacers in those sausages. Quinua is a starchy seed with high nutritive value, which contains substances of technological interest in dry-cured manufacturing. The aim of this study was to assess the effect of replacing fat by quinoa on the quality characteristics of a small diameter dry-cured sausage. Three types of sausages were prepared: a control (C; no fat replacement; 30% of pork back-fat), a quinoa half-fat (50% of fat replacement; 15% of pork back-fat), and a quinoa low-fat (LF; 85% of fat replacement; 4.5% of pork back-fat) sausage. Sausages were analyzed for proximate and microbial composition, volatile compounds, and instrumental texture and color. Descriptive and hedonic sensory analyses were also performed. Fat reduction resulted in higher aw , protein content, hardness, chewiness and redness values and spice-derived volatile levels, and in lower cohesiveness values (P < 0.05). Furthermore, the descriptive sensory analysis showed a higher pungent flavor and lower juiciness in LF sausages than in C sausages (P < 0.05). In spite of those differences, fat reduction did not result in a decreased overall acceptance of the sausages by consumers.

Nitrogen oxides and ozone in Portugal: trends and ozone estimation in an urban and a rural site.

This study provides an analysis of the spatial distribution and trends of NO, NO2 and O3 concentrations in Portugal between 1995 and 2010. Furthermore, an estimation model for daily ozone concentrations was developed for an urban and a rural site. NO concentration showed a significant decreasing trend in most urban stations. A decreasing trend in NO2 is only observed in the stations with less influence from emissions of primary NO2. Several stations showed a significant upward trend in O3 as a result of the decrease in the NO/NO2 ratio. In the northern rural region, ozone showed a strong correlation with wind direction, highlighting the importance of long-range transport. In the urban site, most of the variance is explained by the NO2/NOX ratio. The results obtained by the ozone estimation model in the urban site fit 2013 observed data. In the rural site, the estimated ozone during extreme events agrees with observed concentration.

Screening and Biological Effects of Marine Pyrroloiminoquinone Alkaloids: Potential Inhibitors of the HIF-1α/p300 Interaction.

Inhibition of the hypoxia-inducible factor 1α (HIF-1α) pathway by disrupting its association with the transcriptional coactivator p300 inhibits angiogenesis and tumor development. Development of HIF-1α/p300 inhibitors has been hampered by preclinical toxicity; therefore, we aimed to identify novel HIF-1α/p300 inhibitors. Using a cell-free assay designed to test compounds that block HIF-1α/p300 binding, 170 298 crude natural product extracts and prefractionated samples were screened, identifying 25 active extracts. One of these extracts, originating from the marine sponge Latrunculia sp., afforded six pyrroloiminoquinone alkaloids that were identified as positive hits (IC50 values: 1-35 µM). Luciferase assays confirmed inhibition of HIF-1α transcriptional activity by discorhabdin B (1) and its dimer (2), 3-dihydrodiscorhabdin C (3), makaluvamine F (5), discorhabdin H (8), discorhabdin L (9), and discorhabdin W (11) in HCT 116 colon cancer cells (0.1-10 µM, p < 0.05). Except for 11, all of these compounds also reduced HIF-1α transcriptional activity in LNCaP prostate cancer cells (0.1-10 µM, p < 0.05). These effects occurred at noncytotoxic concentrations (<50% cell death) under hypoxic conditions. At the downstream HIF-1α target level, compound 8 (0.5 µM) significantly decreased VEGF secretion in LNCaP cells (p < 0.05). In COLO 205 colon cancer cells no activity was shown in the luciferase or cytotoxicity assays. Pyrroloiminoquinone alkaloids are a novel class of HIF-1α inhibitors, which interrupt the protein-protein interaction between HIF-1α and p300 and consequently reduce HIF-related transcription.

Stelliosphaerols A and B, Sesquiterpene-Polyol Conjugates from an Ecuadorian Fungal Endophyte.

Endophytic fungi are plant tissue-associated fungi that represent a rich resource of unexplored biological and chemical diversity. As part of an ongoing effort to characterize Amazon rainforest-derived endophytes, numerous fungi were isolated and cultured from plants collected in the Yasuní National Park in Ecuador. Of these samples, phylogenetic and morphological data revealed a previously undescribed fungus in the order Pleosporales that was cultured from the tropical tree Duroia hirsuta. Extracts from this fungal isolate displayed activity against Staphylococcus aureus and were thus subjected to detailed chemical studies. Two compounds with modest antibacterial activity were isolated, and their structures were elucidated using a combination of NMR spectroscopic analysis, LC-MS studies, and chemical degradation. These efforts led to the identification of stelliosphaerols A (1) and B (2), new sesquiterpene-polyol conjugates that are responsible, at least in part, for the S. aureus inhibitory activity of the fungal extract.

neo-Clerodane Diterpenoids from Ajuga macrosperma var. breviflora.

Six new naturally occurring ajugarin-like neo-clerodane diterpenoids, ajugaflorins A-F, along with six known compounds [the parent ajugarin I, ajugalides B and C, ajugamarin F4, ajugamacrin E, and ajugatakasin B] were isolated from A. macrosperma var. breviflora. The structures were elucidated by extensive NMR spectroscopic and MS analyses and comparison with data previously reported.

Indoor aerosol size distributions in a gymnasium.

In this study, an indoor/outdoor monitoring program was carried out in a gymnasium at the University of Leon, Spain. The main goal was a characterization of aerosol size distributions in a university gymnasium under different conditions and sports activities (with and without magnesia alba) and the study of the mass fraction deposited in each of the parts of the respiratory tract. The aerosol particles were measured in 31 discrete channels (size ranges) using a laser spectrometer probe. Aerosol size distributions were studied under different conditions: i) before sports activities, ii) activities without using magnesia alba, iii) activities using magnesia alba, iv) cleaning procedures, and v) outdoors. The aerosol refractive index and density indoors were estimated from the aerosol composition: 1.577-0.003i and 2.055 g cm(-3), respectively. Using the estimated density, the mass concentration was calculated, and the evolution of PM1, PM2.5 and PM10 for different activities was assessed. The quality of the air in the gymnasium was strongly influenced by the use of magnesia alba (MgCO3) and the number of gymnasts who were training. Due to the climbing chalk and the constant process of resuspension, average PM10 concentrations of over 440 µg m(-3) were reached. The maximum daily concentrations ranged from 500 to 900 µg m(-3). Particle size determines the place in the respiratory tract where the deposition occurs. For this reason, the inhalable, thoracic, tracheobronchial and respirable fractions were assessed for healthy adults and high risk people, according to international standards. The estimations show that, for healthy adults, up to 300 µg m(-3) can be retained by the trachea and bronchi, and 130 µg m(-3) may reach the alveolar region. The different physical activities and the attendance rates in the sports facility have a significant influence on the concentration and size distributions observed.

Particulate matter in the indoor and outdoor air of a gymnasium and a fronton.

An indoor/outdoor monitoring programme of PM10 was carried out in two sports venues (a fronton and a gymnasium). Levels always below 50 µg m(-3) were obtained in the fronton and outdoor air. Due to the climbing chalk and the constant process of resuspension, concentrations above 150 µg m(-3) were registered in the gymnasium. The chalk dust contributed to CO3 (2-) concentrations of 32 ± 9.4 µg m(-3) in this sports facility, which represented, on average, 18 % of the PM10 mass. Here, the carbonate levels were 128 times higher than those registered outdoors. Much lower concentrations, around 1 µg m(-3), were measured in the fronton. The chalk dust is also responsible for the high Mg(2+) concentrations in the gym (4.7 ± 0.89 µg m(-3)), unfolding a PM10 mass fraction of 2.7 %. Total carbon accounted for almost 30 % of PM10 in both indoor spaces. Aerosol size distributions were bimodal and revealed a clear dependence on physical activities and characteristics of the sports facilities. The use of climbing chalk in the gymnasium contributed significantly to the coarse mode. The average geometric mean diameter, geometric standard deviation and total number of coarse particles were 0.77 µm, 2.79 cm(-3) and 28 cm(-3), respectively.

neo-Clerodane diterpenoids from Ajuga bracteosa.

Different neo-clerodane diterpenoids were isolated from a dichloromethane extract of Ajuga bracteosa depending on the isolation procedure used, owing to the labile nature of these tetrahydrofurofuran derivatives. Under "hydroxyl-free" purification conditions, both clerodin- and dihydroclerodin-type diterpenes were obtained [four new compounds, ajubractins A-D (1-4), along with clerodin (5), 3-epi-caryoptin (6), ajugapitin (7), 14,15-dihydroclerodin (8), 3-epi-14,15-dihydrocaryoptin (9), ivain II (10), and 14,15-dihydroajugapitin (11)]. When methanol-water mixtures were used for a C18 reversed-phase prepurification procedure and for semipreparative HPLC, the new ajubractin E (12) was also isolated along with 3 and 8-11, as previously, but 7 was the only tetrahydrofurofuran derivative obtained. Epimeric (15R and 15S) mixtures were obtained instead of 14-hydro-15-hydroxyclerodin derivatives [15-hydroxyajubractin C (13), 14-hydro-15-hydroxyajugachin A (14), and 14-hydro-15-hydroxyajugapitin (15)], along with 15-epi-lupulin B (16). The structures of the new compounds were elucidated by NMR and MS data analysis and by comparison with values previously reported. Antifeedant activity against Spodoptera littoralis larvae was evaluated for the compounds obtained.

Phytotoxic activity of flavonoids from Dicranostyles ampla.

Crude extracts from over 16 species of plants from the family Convolvulaceae were evaluated for phytotoxic activity against Agrostis stolonifera (bentgrass) and Lactuca sativa (lettuce) at 1000 microg/mL. Ethanol extracts of Dicranostyles ampla Ducke were among the most active of those species tested. Systematic bioassay-guided fractionation of the ethanol extract of the aerial parts from this species was performed to identify specifically the phytotoxic compounds. Two phytotoxic flavonoids, dihydromyricetin (1) and myricetin-3-O-alpha-rhamnoside or myricetrin (2), were found to be responsible for much of the activity of the extract as a whole in the A. stolonifera and L. sativa bioassay. In a Lemna paucicostata bioassay, 1 and 2 had no activity at 100 microM.

LC-MS-SPE-NMR for the isolation and characterization of neo-clerodane diterpenoids from Teucrium luteum subsp. flavovirens (perpendicular).

neo-Clerodane diterpenes of plant origin are molecules difficult to monitor due to their nonspecific UV/vis absorption. The present work describes for the first time the application of the LC-MS-SPE-NMR technique for the isolation and characterization of three new neo-clerodane diterpenes, 3beta-hydroxyteucroxylepin and teuluteumin A and teuluteumin B, from Teucrium luteum subsp. flavovirens, harvested from two different locations.

Phytoecdysteroids from Ajuga macrosperma var. breviflora roots.

Three new phytoecdysteroids, ajugacetalsterones C (1) and D (3) and breviflorasterone (2), were isolated from the roots of Ajuga macrosperma var. breviflora along with five known compounds, namely, 20-hydroxyecdysone, cyasterone, makisterone A, 20-hydroxyecdysone 3-acetate, and 20-hydroxyecdysone 2-acetate. The structures of 1-3 were elucidated on the basis of extensive 1D and 2D NMR spectroscopic studies. The new compounds possess acetal oxygen bridges between C-26 and C-20/C-22, or C-26/C-23, or a lactone bridge between C-26 and C-23.