Bioactivity Screening of Antarctic Sponges Reveals Anticancer Activity and Potential Cell Death via Ferroptosis by Mycalols

Sponges are known to produce a series of compounds with bioactivities useful for human health. This study was conducted on four sponges collected in the framework of the XXXIV Italian National Antarctic Research Program (PNRA) in November-December 2018, i.e., Mycale (Oxymycale) acerata, Haliclona (Rhizoniera) dancoi, Hemimycale topsenti, and Hemigellius pilosus. Sponge extracts were fractioned and tested against hepatocellular carcinoma (HepG2), lung carcinoma (A549), and melanoma cells (A2058), in order to screen for antiproliferative or cytotoxic activity. Two different chemical classes of compounds, belonging to mycalols and suberitenones, were identified in the active fractions. Mycalols were the most active compounds, and their mechanism of action was also investigated at the gene and protein levels in HepG2 cells. Of the differentially expressed genes, ULK1 and GALNT5 were the most down-regulated genes, while MAPK8 was one of the most up-regulated genes. These genes were previously associated with ferroptosis, a programmed cell death triggered by iron-dependent lipid peroxidation, confirmed at the protein level by the down-regulation of GPX4, a key regulator of ferroptosis, and the up-regulation of NCOA4, involved in iron homeostasis. These data suggest, for the first time, that mycalols act by triggering ferroptosis in HepG2 cells.     

Stress-induced biosynthesis of dicaffeoylquinic acids in globe artichoke

Leaf extracts from globe artichoke ( Cynara cardunculus L. var. scolymus) have been widely used in medicine as hepatoprotectant and choleretic agents. Globe artichoke leaves represent a natural source of phenolic acids with dicaffeoylquinic acids, such as cynarin (1,3-dicaffeoylquinic acid), along with its biosynthetic precursor chlorogenic acid (5-caffeoylquinic acid) as the most abundant molecules. This paper reports the development of an experimental system to induce caffeoylquinic acids. This system may serve to study the regulation of the biosynthesis of (poly)phenolic compounds in globe artichoke and the genetic basis of this metabolic regulation. By means of HPLC-PDA and accurate mass LC-QTOF MS and MS/MS analyses, the major phenolic compounds in globe artichoke leaves were identified: four isomers of dicaffeoylquinic acid, three isomers of caffeoylquinic acid, and the flavone luteolin 7-glucoside. Next, plant material was identified in which the concentration of phenolic compounds was comparable in the absence of particular treatments, with the aim to use this material to test the effect of stress application on the regulation of biosynthesis of caffeoylquinic acids. Using this material, the effect of UV-C, methyl jasmonate, and salicylic acid treatments on (poly)phenolic compounds was tested in different globe artichoke genotypes. UV-C exposure consistently increased the levels of dicaffeoylquinic acids in all genotypes, whereas the effect on compounds from the same biosynthetic pathway, for example, chlorogenic acid and luteolin-7-glucoside, was much less pronounced and was not statistically significant. No effect of methyl jasmonate or salicylic acid was found. Time-response experiments indicated that the level of dicaffeoylquinic acids reached a maximum at 24 h after UV radiation. On the basis of these results a role of dicaffeoylquinic acids in UV protection in globe artichoke is hypothesized.     

Adenocarcinoma of apocrine sweat glands in a mouflon (Ovis musimon).

A free-living mouflon (Ovis musimon) was presented with a mass on the left shoulder. At necropsy, multifocal, slightly protruding whitish spots were noted on the kidneys, and several lymph nodes were abnormal. Histologically, the mass was composed of epithelial cells arranged in tubular and tubulopapillary structures. The cytoplasm of the epithelial cells had numerous periodic acid-Schiff-positive and diastase-resistant granules. Ultrastructurally, the cytoplasm of the neoplastic cells contained numerous pleomorphic secretory granules and microvilli, which partially covered the luminal surface of the tumor cells. Metastatic foci were present in prescapular and mediastinal lymph nodes and kidneys. On the basis of histological and ultrastructural findings, this tumor was diagnosed as a tubulopapillary adenocarcinoma, arising from apocrine sweat glands of the skin.     

Detection of pathogens in ovine and caprine abortion samples from Sardinia, Italy, by PCR.

During 2003-2005, 399 abortion samples (315 fetuses and 84 placentae) were collected from 107 ovine and caprine farms in northern Sardinia. Tissues from aborted fetuses and placentae were examined by PCR assay to detect DNA from Coxiella burnetii, Chlamydophila abortus, Salmonella enterica Serovar abortusovis, Toxoplasma gondii, and Neospora caninum. The DNA from at least 1 of these 5 infectious agents was amplified in 41% of ovine fetuses, while only 17% of the caprine fetuses yielded a positive amplification result for at least 1 of the 5 agents. Out of a total of 366 ovine aborted samples, T. gondii DNA was detected most frequently (18.1% of fetuses and 13.1% of placentae), followed by S. abortusovis (13% of fetuses and 14.4% of placentae), C. burnetii (10.9% of fetuses, of 9.2% placentae), C. abortus (2.4% of fetuses, 6.5% of placentae), and N. caninum (2% of placentae). In 33 fetuses and 9 placentae, the simultaneous presence of pathogens with different associations was detected. Out of a total of 31 caprine aborted samples, T. gondii was detected most frequently (13% of fetuses and 25% of placentae), followed by C. abortus (12.5% of placentae), C. burnetii (12.5% of placentae), and N. caninum (8.6%).     

Chemical composition and antimicrobial activity of the essential oil from Ferula glauca L. (F. communis L. subsp. glauca) growing in Marche (central Italy)

The essential oil obtained from different parts of Ferula glauca L. (formerly considered as a subspecies of F. communis) growing in Marche (central Italy), was analyzed for the first time by GC-FID and GC-MS. The major volatiles were (E)-caryophyllene and caryophyllene oxide in leaves, alpha-pinene, myrcene and germacrene D in flowers, alpha- and beta-pinene in fruits, (E)-beta-farnesene, myristicin and elemicin in roots, respectively. The differences in composition detected with respect to F. communis, made the volatile fraction a reliable marker to distinguish between them, and confirm the botanical data at the base of their discrimination. Furthermore, the oil was assayed for its antimicrobial activity by the broth microdilution method. B. subtilis was found to be the most sensitive microorganism, with the lowest MIC values.     

SSR-based identification key of cultivars of Olea europaea L. diffused in Southern-Italy

Olive (Olea europaea L.) is an important fruit species in Italy and Mediterranean basin constituted by a wide germplasm with a large number of cultivars present in all the Mediterranean area. SSRs are becoming the markers of choice for variability studies in olives as they are simple to perform, transferable, hypervariable, highly polymorphic and show a high information content.Olive genetic diversity was studied using 16 SSR markers on 30 cultivars diffused in Southern-Italy. Resolving Power (RP) and Power of Discrimination (PD) were calculated to evaluate the efficiency of the SSR markers investigated in studies of cultivars fingerprinting. Based on their high efficiency, two SSR markers, UDO43 and DCA16 were chosen to set up an identification key to distinguish the entire set of cultivars, confirming the high biodiversity of the Southern-Italian olive germplasm and the suitability of SSR markers in studies of cultivar diagnosis.     

Short-term impact of spent coffee grounds over soil organic matter composition and stability in two contrasted Mediterranean agricultural soils

Purpose

Spent coffee grounds (SCG) is a biowaste which arouse great interest as soil organic amendment due to the huge amount produced around the world. However, the impact of this residue on soil organic matter (SOM) functionality and stability has been barely studied. Thus, the aim of this work is to study the short-term effects of SCG on the quantity and quality of SOM in two Mediterranean agricultural soils (Vega soil, SV and Red soil, SR) in microcosm conditions.

Materials and methods

The in vitro assay was performed with two fresh SCG doses (2.5 and 10% w/w), two incubation times (30 and 60 days) and two agricultural soils (SV and SR). SOM fractionation to obtain total extractable carbon, humic acids, fulvic acids, humins and hot water soluble carbon (HWSC) was determined. Spectroscopic UV-Vis and Mid-IR, thermogravimetric and simultaneous differential thermal analysis, as well as scanning electron microscopy (SEM), were also applied in this study.

Results and discussion

SCG increased all SOM fractions, especially the levels of more labile SOM (HWSC, increased 600–700%) and total extractable carbon (increased to around 200%). SCG also increased humic acids and fulvic acids around 200%, but the functionality of humic acids was affected by a reduction of the functional groups with more recalcitrant and stable character. The tested soils are different from each other (the SV has a more clayish texture and a higher smectite clay content than the SR) which made the behaviour of these soils different. The degree of incorporation of SCG into the soils structure and the interaction between soil and SCG particles (observed by SEM) affected carbon retention under stable forms, increasing carbon stabilization in SV with respect to SR.

Conclusions

The short-term effect of SCG on SOM composition and functionality demonstrate that this bioresidue could be used as soil organic amendment, being a valuable alternative use of a polluting waste. Soil type is a key factor since it influences the soil-SCG interaction and consequently SOM stability. To deepen the study of those effects, it would be necessary to analyze the long-term effects, field studies and to test in a greater number of soil types.

     

Impacts of rainstorms on soil erosion and organic matter for different cover crop systems in the western coast agricultural region of Syria

More and more cultivated coastal territories of Syria are being affected by increasing land degradation processes, specifically, by soil erosion due to non‐sustainable soil management. The use of cover crops can be considered an ideal solution to reduce the negative impacts of extreme rainfall events on soil erosion. However, there is no enough information about the main cultivated areas of Syria due to the current conflicts and the very few types of research conducted there. Therefore, the main aim of this research was to assess soil erosion (soil loss and sediment concentration) and hydrological response (run‐off and infiltration) considering the impact of different types of cropping systems on soil organic matter. To achieve this goal, using erosion plots, five grouped natural rainfall events and soil samples were collected, and infiltration measurements were performed between November 2012 and April 2013 (rainy season) on three different cropping systems (wheat, vetch, bean and control) with different sloping gradients (8% and 20%) in Tartous governorate (W Syria). Our results showed a cumulative rainfall amount from 68.8 to 201 mm during the study period. The highest sediment yield was generated in the control and wheat plots, registering 0.468 and 0.368 kg m^?2, respectively, with an inclination of 8%. For an inclination of 20%, the control and wheat plots registered 1.145 and 0.967 kg m^?2, respectively. In comparison with the control plots, there was a decrease in the total eroded organic matter, which ranged from 0.0579 (control) to 0.0289 (bean) kg m^?2 year^?1 in the plots with 20% inclination, and from 0.0233 (control) to 0.0069 (bean) kg m^?2 year^?1 with 8% inclination. To sum up, bean and vetch play an effective role to mitigate soil erosion, delay run‐off and reduce sediment yield. The output of this research provides first insights into the impact of different land uses on soil loss. Also, it could help rural inhabitants and farmers to correctly manage their soils against soil erosion processes.     

An integrated geochemical and mineralogical approach for the evaluation of arsenic mobility in mining soils

Purpose  

The assessment of risk related to the presence of potentially toxic elements in soils is strictly related to the knowledge of their form and mobility. These relevant properties depend on the complex interactions of the elements of concern with the soil particles that generally cannot be addressed by a single technique. This study presents an integrated approach implementing geochemical and mineralogical investigation techniques on samples from a former mining area (Tolfa Mountains district, northern Latium, Italy), where exploiting activities occurred until the recent past. In particular, the As total concentration and the As distribution in solid phases is studied with the aim to evaluate the possibility of environmental pollution and consequent risks for the health of people living in the area and possibly affected in case of significant mobilization of this toxic element.