S-100 Fraction Research Articles

Phytochemical Composition and Cytoprotective Properties of the Endemic Sideritis sipylea Boiss Greek Species: A Valorization Study.

Sideritis sipylea Boiss. (Fam. Lamiaceae) is an endemic plant of the North Aegean Islands (Greece), commonly known as ironwort. Traditionally, its aerial parts have been used to relieve several ailments, especially gastrointestinal disorders, however, with scant knowledge about the pharmacological basis. In the present study, an endemic S. sipylea Greek species from Lesvos Island has been characterized for phytochemical composition and biological activities, in order to give a possible scientific basis to its traditional use and to highlight a further nutraceutical interest as a source of bioactive phytochemicals and extracts. Three different fractions obtained from a methanolic extract of S. sipylea aerial parts by using ethyl acetate with 10 (S10), 20 (S20), and 50% (S50) methanol as fractionation solvents were phytochemically characterized. Moreover, their antioxidant power and cytoprotective activity in different human cell lines were evaluated. The phytochemical analysis highlighted the presence of flavonoids, iridoids, and phenolic acids in all the tested samples. Particularly, the S10 fraction mainly contained iridoids, while S20 and S50 lavandulifolioside and chlorogenic acid, respectively. The fractions also showed antioxidant properties, S10 and S20 being the most potent. When assessed in human cholangiocytes, they counteracted the cytotoxicity of the tBOOH pro-oxidant agent, by reducing ROS levels and affecting GSH antioxidant system. The present findings highlight a possible interest in S10 and S20 fractions from S. sipylea as sources of bioactive molecules and stimulate further studies in order to characterize their possible application for nutraceutical and pharmaceutical purposes.

Purification and identification of novel antioxidant peptides from silkworm pupae (Bombyx mori) protein hydrolysate and molecular docking study

Crude proteins from the pupae of two indigenous strains of silkworm (Bombyx mori), Nangnoi and Nangsew were hydrolyzed by alcalase, papain and trypsin. Protein hydrolysate of Nangnoi strain prepared by alcalase showed the highest antioxidant activities. In addition, the DNA damage protection ability of this protein hydrolysate was evaluated. Subsequently, this protein hydrolysate was purified by passing through a 3 kDa MW cut-off membrane followed by reverse phase high performance liquid chromatography (RP-HPLC). Amongst eight fractions obtained after RP-HPLC purification, fraction S3 showed interesting antioxidant activities with the IC50 values toward ABTS and DPPH radicals of 4.78 ± 0.08 and 7.83 ± 0.02 μg/mL, respectively. Potentially antioxidant peptides, AAEYPA and AKPGVY were identified in the S3 fraction after analysis by LC-MS/MS. ABTS and DPPH radicals scavenging activities of both synthesized peptides were investigated. These results suggest that both hexapeptides are potential antioxidants that can be used in the food and cosmetic industries.

Molecular Weight Identification of Compounds Involved in the Fungal Synthesis of AgNPs: Effect on Antimicrobial and Photocatalytic Activity.

The biological synthesis of silver nanoparticles (AgNPs) for medical, environmental, and industrial applications is considered an alternative to chemical synthesis methods. Additionally, the reducing, capping, and stabilizing molecules produced by the organisms can play a key role in the further activity of AgNPs. In this work, we evaluated the synthesis of AgNPs by four molecular weight fractions (S1: <10 kDa, S2: 10 to 30 kDa, S3: 30 to 50 kDa, and S4: >50 kDa) of mycelia-free aqueous extract produced by the white-rot fungus Stereum hirsutum and their effect on the antimicrobial activity against Pseudomonas syringae and photocatalytic decolorization of nine synthetic dyes exposed to sunlight radiation. All synthesis assay fractions showed the characteristic surface plasmon resonance (SPR) with 403 to 421 nm peaks. TEM analysis of synthesized AgNPs showed different sizes: the whole mycelia-free extracts S0 (13.8 nm), S1 (9.06 nm), S2 (10.47 nm), S3 (22.48 nm), and S4 (16.92 nm) fractions. The results of disk diffusion assays showed an inverse relation between antimicrobial activity and the molecular weight of compounds present in the mycelia-free aqueous extract used to synthesize AgNPs. The AgNPs synthesized by S0 (14.3 mm) and S1(14.2 mm) generated the highest inhibition diameter of P. syringae growth. By contrast, in the photocatalytic assays, the AgNPs synthesized by the S2 fraction showed the highest discoloration in all the dyes tested, reaching 100% of the discoloration of basic dyes after 2 h of sunlight exposure. The maximum discoloration observed in reactive and acid dyes was 53.2% and 65.3%, respectively. This differentiation in the antimicrobial and photocatalytic activity of AgNPs could be attributed to the capping effect of the molecules present in the extract fractions. Therefore, the molecular separation of synthesis extract enables the specific activities of the AgNPs to be enhanced.

Novel Process of Reduction Roasting Manganese Ore with Sulfur Waste and Extraction of Mn by Acid Leaching

Manganese dioxide is typically reduced to a bivalent state before being extracted; here, sulfur is considered an efficient reductant and sulfur–based reduction has been industrialized in China. In this study, the reaction mechanism between MnO2 and gaseous sulfur was investigated. Thermodynamically, the reduction of MnO2 by gaseous sulfur is feasible. The predominant phase diagram as functions of temperature and input S2(g) fraction in the S2–MnO2 system was calculated. Experimental validation showed that MnO2 was reduced stepwise to low-valence manganese oxides and manganese sulfate. The phase composition of the roasted products was complex, and MnS was inevitably formed. The valence state as well as microstructure of manganese dioxide during reduction roasting were also investigated by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy–energy-dispersive spectroscopy (SEM–EDS). The reaction process could be described by an unreacted nuclear model. Manganese was extracted by sulfuric acid solution after reduction by sulfur waste. In sulfuric acid, 95.2 wt% Mn extraction was achieved, using a roasting temperature of 450 °C, roasting time of 30 min, and S2/MnO2 molar ratio of 0.40. With the same conditions, low Fe extraction was achieved. On the other hand, in deionized water, 24.3 wt% Mn extraction was achieved, confirming the formation of MnSO4.

Cytotoxicity and Apoptosis Induction of 6,7-Dehydroroyleanone from Taiwania cryptomerioides Bark Essential Oil in Hepatocellular Carcinoma Cells.

The objective of the present study is to evaluate the cytotoxicity of Taiwania cryptomerioides essential oil and its phytochemical on the Hep G2 cell line (human hepatocellular carcinoma). Bark essential oil has significant cytotoxicity to Hep G2 cells, and S3 fraction is the most active fraction in cytotoxicity to Hep G2 cells among the six fractions. The diterpenoid quinone, 6,7-dehydroroyleanone, was isolated from the active S3 fraction by bioassay-guided isolation. 6,7-Dehydroroyleanone exhibited significant cytotoxicity in Hep G2 cells, and the efficacy of 6,7-dehydroroyleanone was better than the positive control, etoposide. Apoptosis analysis of Hep G2 cells with different treatments was characterized via flow cytometry to confirm the cell death situation. Etoposide and 6,7-dehydroroyleanone could induce the apoptosis in Hep G2 cells using flow cytometric assay. Results revealed 6,7-dehydroroyleanone from T. cryptomerioides bark essential oil can be a potential phytochemical to develop the anticancer chemotherapeutic agent for the treatment of the human hepatocellular carcinoma.

Latitudinal variability of preserved sedimentary organic matter along the Peruvian continental margin as inferred from petrographic and geochemical properties

Processes controlling preservation and accumulation of organic matter (OM) in marine sediments are highly sensitive towards both natural and anthropogenic impacts. Thus, the sedimentary OM is considered a key proxy for the characterization of recent and past sedimentary environments. Along the Peruvian continental margin (PCM), high primary production and an intense oxygen minimum zone (OMZ) favor the accumulation and preservation of OM in sediments. Spatial variations in oceanographic conditions impact on both productivity and the redox conditions, in particular the OMZ intensity. These factors in turn lead to spatial differences in depositional conditions and OM accumulation and preservation. Therefore, this study aims at characterizing nature and types of recent sedimentary OM along the PCM covering a wide latitudinal and bathymetric range. With this purpose, a total of 29 surface sediment samples were collected from Tumbes (3°S) to San Juan (15°S) and analyzed by means of organic petrography (palynofacies analysis) and Rock-Eval pyrolysis. Results allowed characterizing the OM from recent sediments. Along the PCM, the sedimentary OM is predominantly amorphous, revealing its overwhelming marine origin. The highest abundances of gelified amorphous OM, total organic carbon, S1 and S2 Rock-Eval fractions occurred towards the southern part of the study area, especially off Callao (12°S) and Pisco (14°S), considered important upwelling and primary productivity centers. A principal component analysis performed allowed the description of main OM depositional environments. In the outer continental shelf of Callao (12°S) and OMZ of Pisco (14°S), accumulation of well-preserved OM occurs whereas in the inner shelf of Callao redox oscillations lead to less OM accumulation and pyrite formation in sediments. North of Callao, from Tumbes (3°S) to Huacho (11°S), a mixed signal of OM accumulation was observed. Finally, along the slope, transportation processes seem to play a key role in OM accumulation. This study also allowed the evaluation of certain proxies (e.g. Rock-Eval hydrogen index and oxygen index) routinely used in palaeoceanographic studies.

Structural and catalytic roles of the human 18S rRNA methyltransferases DIMT1 in ribosome assembly and translation

rRNA-modifying enzymes participate in ribosome assembly. However, whether the catalytic activities of these enzymes are important for the ribosome assembly and other cellular processes is not fully understood. Here, we report the crystal structure of WT human dimethyladenosine transferase 1 (DIMT1), an 18S rRNA N6,6-dimethyladenosine (m26,6A) methyltransferase, and results obtained with a catalytically inactive DIMT1 variant. We found that DIMT1+/- heterozygous HEK 293T cells have a significantly decreased 40S fraction and reduced protein synthesis but no major changes in m26,6A levels in 18S rRNA. Expression of a catalytically inactive variant, DIMT1-E85A, in WT and DIMT1+/- cells significantly decreased m26,6A levels in 18S rRNA, indicating a dominant-negative effect of this variant on m26,6A levels. However, expression of the DIMT1-E85A variant restored the defects in 40S levels. Of note, unlike WT DIMT1, DIMT1-E85A could not revert the defects in protein translation. We found that the differences between this variant and the WT enzyme extended to translation fidelity and gene expression patterns in DNA damage response pathways. These results suggest that the catalytic activity of DIMT1 is involved in protein translation and that the overall protein scaffold of DIMT1, regardless of the catalytic activity on m26,6A in 18S rRNA, is essential for 40S assembly.

1973-P: Single-Nuclei Transcriptomics of Human Adipose Tissue Identify Distinct Adipocyte Progenitor Subpopulations in Type 2 Diabetes

Altered adipocyte function underlies type 2 diabetes (DM) pathogenesis. We hypothesized that adipocyte dysfunction in visceral adipose tissue (VAT) in DM originates from distinct adipocyte progenitor cell (APC) subpopulations. VAT was collected from bariatric surgery patients with (HbA1c&amp;gt;6.5) and without DM (NDM, HbA1c&amp;lt;5.7). Single-nuclei RNA sequencing was performed in VAT nuclei (n= 1 NDM, 2 DM) using 10X Genomics. Transcriptionally distinct cell subpopulations were identified by DEG: mature adipocytes (ADIPOQ, LEP), endothelial cells (VWR), immune cells (F13A1, MSR1), and two major APC subpopulations (S1: FABP4, ADIRF, ITLN1 and MSLN; S2: APOD, CFD and WISP2). S2 was uniquely identified by the upregulation of extracellular matrix accumulation and fibrosis markers: LUM, DCN, FBN1, and COL1A1, 3A1, 6A1 and 6A3. S2 had higher expression of APC and committed preadipocyte markers, such as PLIN1, GNAI1, CXCL14, CP, and CRISPLD2, while S1 had higher mesothelial-like cell markers GPM6A, PKHD1L1, LRRN4, CXADR, SLPI, and UPK3B. We FACS-isolated S1 and S2 from VAT CD31-CD45- cells (n= 3 NDM, 5 DM) using the surface marker TM4SF1, which was abundantly expressed in S1, but not S2. TM4SF1 expression (qPCR) was 50% higher in S1 (TM4SF1+) than in S2 (TM4SF1-). TM4SF1 immunostaining of human SVF confirmed distinct APC subpopulations. S1 corresponded to 15 ± 3% and S2 to 69 ± 2% of CD31-CD45- cells. In DM compared to NDM, S2 fraction was increased by 12% while S1 was decreased by 45%. We tested the metabolic function of S1 and S2 sorted from 5 DM VAT. Both subpopulations differentiated into lipid-filled adipocytes upon adipogenic induction in vitro. However, S2 compared to S1, manifested decreased proliferation and increased intracellular lipid accumulation and lipolytic response, consistent with a pro-hypertrophic, lipolytic adipocyte phenotype in DM. Our data suggest that distinct APC subpopulations in human obese VAT contribute to adipose tissue dysfunction in DM. Disclosure C. Strieder-Barboza: None. C.G. Flesher: None. L.M. Geletka: None. R.W. ORourke: None. C.N. Lumeng: None. Funding National Institute of Diabetes and Digestive and Kidney Diseases (R01DK115190, DK090262)

Distinct Microglial Responses in Two Transgenic Murine Models of TAU Pathology.

Microglial cells are crucial players in the pathological process of neurodegenerative diseases, such as Alzheimer’s disease (AD). Microglial response in AD has been principally studied in relation to amyloid-beta pathology but, comparatively, little is known about inflammatory processes associated to tau pathology. In the hippocampus of AD patients, where tau pathology is more prominent than amyloid-beta pathology, a microglial degenerative process has been reported. In this work, we have directly compared the microglial response in two different transgenic tau mouse models: ThyTau22 and P301S. Surprisingly, these two models showed important differences in the microglial profile and tau pathology. Where ThyTau22 hippocampus manifested mild microglial activation, P301S mice exhibited a strong microglial response in parallel with high phospho-tau accumulation. This differential phospho-tau expression could account for the different microglial response in these two tau strains. However, soluble (S1) fractions from ThyTau22 hippocampus presented relatively high content of soluble phospho-tau (AT8-positive) and were highly toxic for microglial cells in vitro, whereas the correspondent S1 fractions from P301S mice displayed low soluble phospho-tau levels and were not toxic for microglial cells. Therefore, not only the expression levels but the aggregation of phospho-tau should differ between both models. In fact, most of tau forms in the P301S mice were aggregated and, in consequence, forming insoluble tau species. We conclude that different factors as tau mutations, accumulation, phosphorylation, and/or aggregation could account for the distinct microglial responses observed in these two tau models. For this reason, deciphering the molecular nature of toxic tau species for microglial cells might be a promising therapeutic approach in order to restore the deficient immunological protection observed in AD hippocampus.

Selective fractionation and isolation of allelopathic compounds from Helianthus annuus L. leaves by means of high-pressure techniques

The allelopathic potential of Helianthus annuus L. leaves was study based on bio-directed chemical fractionation approach. Aerial parts of H. annuus were extracted by means of SFE using supercritical carbon dioxide (scCO2) and ESE using CO2+50% EtOH/H2O (varying ethanol in water from 0 to 100%). Extractions were carried out at 400 bar, 55 °C, 20 g/min and for 4 h. Then, extracts were fractionated in three separators at the following conditions: S1: 200 bar/45 °C; S2: 90 bar/40 °C; and S3: 1 atm/30 °C. ESE obtained higher overall yields than scCO2 and the use of water as cosolvent (CO2+50% H2O) resulted in a S3 fraction free from chlorophylls and rich in bioactive compounds. 14 compounds, including fatty acids, terpenes, flavonoids and heliannuols, were isolated from this fraction. After performing the bioassay on pure compounds, heliannuol D, tambulin, pinoresinol and sesquiterpene 10-oxo-isodauc-3-en-15-al showed the most effective inhibitor profiles.

Numerical study of the burner parameters on the thermal field in a sulfur recovery unit thermal reactor

A sulfur recovery unit (SRU) thermal reactor is the most important equipment in a sulfur plant and is negatively affected by high temperature operations. In this paper, the effect of burner parameters, including the clearance of the acid gas tip and the inlet air swirler angle, on the thermal field in a SRU thermal reactor are investigated numerically, with the aim to reduce the high temperature inside the thermal reactor and to ensure an acceptable sulfur recovery. The simulation results show that the burner with a smaller clearance of the acid gas tip produces a lower temperature, a lower exit SO2 mole fraction and higher exit S2 and H2S mole fractions. Among the clearancs of the acid gas tip investigated, the horizontal clearance of 152.4mm and vertical clearance of 240mm yield the lowest temperature, exit SO2 mole fraction and highest exit S2, H2S mole fractions. The burner with a smaller inlet air swirler angle produces a higher temperature, a higher exit SO2 mole fraction and lower exit S2 and H2S mole fractions. Among the swirler angles investigated, 60° yields the lowest temperature, exit SO2 mole fraction and highest exit S2 , H2S mole fractions.

Comparative study on the antioxidant activity of peptides from pearl oyster (Pinctada martensii) mantle type V collagen and tilapia (Oreochromis niloticus) scale type I collagen

In this study, Pearl oyster mantle type V collagen (POMC) and tilapia scale type I collagen (TSC) were extracted and hydrolyzed by various proteases in order to obtain peptides. The antioxidant activity of the peptides was investigated by DPPH, hydroxyl radical scavenging experiments and a dynamic digestion model in vitro. The results show that there are significant differences in amino acid composition between POMC and TSC. The collagen peptides obtained from pearl oyster mantle (POMCP) by treating with alkaline protease exhibited higher antioxidant activity than that from tilapia scale (TSCP) treated with papaya protease, and both of them showed greater DPPH and hydroxyl radical scavenging activity than other peptides. After being separated via Sephadex G-25 chromatography, the M1 fraction isolated from POMCP, and the S1 fraction from TSCP with which both had higher molecular weights showed the strongest antioxidant activity than other fractions, and the M1 fraction exhibited stronger antioxidant activity than the S1 fraction in scavenging free-radicals and protecting cells from the oxidation damage. Furthermore, after treating the dynamic digestion system model in vitro, the DPPH and hydroxyl radical scavenging activity of the M1 fraction increased slightly. These results suggest that POMCP exhibits stronger antioxidant activity than TSCP, which means that PMOP may be a good candidate to be a potential natural antioxidant in the food-processing industry.

Effect of cadmium bioavailability in food on its compartmentalisation in carabids

Metals assimilated by organisms are sequestered in various compartments and some forms are more stable than others. Sequestration mechanisms used by invertebrates to detoxify metals and prevent interaction with important biomolecules include metal binding to proteins and other ligands, and storage in inorganic granules. The rate and extent at which metal concentrations in different compartments respond to metal concentrations in food and food characteristics has not received much attention, despite being of great relevance. We performed an experiment on the carabid beetle Pterostichus oblongopunctatus exposed to Cd via food made of ground mealworm (Tenebrio molitor) larvae, either reared on Cd contaminated medium or artificially spiked after grinding with CdCl2 solution. Thus, in both cases we used the same type of food, differing only in the soluble Cd pool available to the predators, represented by P. oblongopunctatus. Subcellular compartmentalisation of Cd into organelles, heat-sensitive and heat-stable proteins (the first supernatant, S1 fraction), cellular debris (the second supernatant, S2 fraction) and metal-rich granules (G fraction) was checked a few times during the contamination (90 d) and decontamination (24 d) phases in a toxicokinetic experiment by using different centrifugation steps. The results showed no effect of the type of food (naturally, Cd-N, vs. artificially contaminated with Cd, Cd-A) on Cd sequestration kinetics in P. oblongopunctatus, but the amount of Cd sequestered in the S1 and G fractions were in general higher in the Cd-A than the Cd-N treatment, indicating that Cd transfer in the food web depends on the speciation of the metal in the food. The proportional distribution of Cd over different fractions was, however, similar in beetles fed both food types. Most of the accumulated Cd in the beetles existed as fraction S1 (ca. 35%), which is important for the transfer of metals to higher trophic levels in a food web.

CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility

Mutations in the Cyclin-dependent kinase-like 5 (CDKL5) gene cause severe neurodevelopmental disorders accompanied by intractable epilepsies, i.e. West syndrome or atypical Rett syndrome. Here we report generation of the Cdkl5 knockout mouse and show that CDKL5 controls postsynaptic localization of GluN2B-containing N-methyl-d-aspartate (NMDA) receptors in the hippocampus and regulates seizure susceptibility. Cdkl5 −/Y mice showed normal sensitivity to kainic acid; however, they displayed significant hyperexcitability to NMDA. In concordance with this result, electrophysiological analysis in the hippocampal CA1 region disclosed an increased ratio of NMDA/α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated excitatory postsynaptic currents (EPSCs) and a significantly larger decay time constant of NMDA receptor-mediated EPSCs (NMDA-EPSCs) as well as a stronger inhibition of the NMDA-EPSCs by the GluN2B-selective antagonist ifenprodil in Cdkl5 −/Y mice. Subcellular fractionation of the hippocampus from Cdkl5 −/Y mice revealed a significant increase of GluN2B and SAP102 in the PSD (postsynaptic density)-1T fraction, without changes in the S1 (post-nuclear) fraction or mRNA transcripts, indicating an intracellular distribution shift of these proteins to the PSD. Immunoelectron microscopic analysis of the hippocampal CA1 region further confirmed postsynaptic overaccumulation of GluN2B and SAP102 in Cdkl5 −/Y mice. Furthermore, ifenprodil abrogated the NMDA-induced hyperexcitability in Cdkl5 −/Y mice, suggesting that upregulation of GluN2B accounts for the enhanced seizure susceptibility. These data indicate that CDKL5 plays an important role in controlling postsynaptic localization of the GluN2B-SAP102 complex in the hippocampus and thereby regulates seizure susceptibility, and that aberrant NMDA receptor-mediated synaptic transmission underlies the pathological mechanisms of the CDKL5 loss-of-function.

Actomyosin interaction at low ATP concentrations.

In vitro motility assay (IVMA) experiments were performed to analyze the movement of actin filaments sliding on a pavement of myosin molecules at different [ATP] and [ADP]. In standard experimental conditions at [ATP]=2mM, about 80% of the actin filaments move in unloaded conditions with a constant velocity. However, a fraction of at least 20% static actin filaments is always present. The accepted explanation is the occurrence of damaged "rigor"-like myosin heads that do not undergo the normal ATP-dependent cycling motion. However, in a series of IVMA experiments performed at different [ATP] we observed that the mobility of actin filaments increased with lowering [ATP]. We investigated the influence of [ATP] on the number of mobile actin filaments. IVMA experiments were performed at controlled nucleotide concentrations and the percentage of mobile filaments accurately determined by specific operator-guided software. The value of ΔG ATP involved was determined. Results showed that the number of mobile actin filaments sliding on type 2B heavy meromyosin isoform (2B HMM) increased at very low [ATP] accompanied by less negative ΔG ATP values. Similar results were obtained by increasing [ADP]. Performing experiments at the same [ATP] with different myosin types, we found a higher number of mobile actin filaments on slow type 1 HMM with respect to type 2B HMM while the highest number of mobile actin filaments was found on single-head myosin (S1 fraction). We also found that [ATP] did not influence the percentage of mobile actin filaments sliding on S1. Our results reveal novel aspects of actomyosin interaction.

Subcellular partitioning of cadmium and zinc in mealworm beetle (Tenebrio molitor) larvae exposed to metal-contaminated flour

By studying the internal compartmentalization of metals in different subcellular fractions we are able to better understand the mechanisms of metal accumulation in organisms and the transfer of metals through trophic chains. We investigated the internal compartmentalization of cadmium (Cd) and zinc (Zn) in mealworm beetle (Tenebrio molitor) larvae by breeding them in flour contaminated with either Cd at 100, 300 and 600mgkg−1, or Zn at 1000 and 2000mgkg−1. We separated the cellular components of the larvae into 3 fractions: the S1 or cytosolic fraction containing organelles, heat-sensitive and heat-stable proteins, the S2 or cellular debris fraction and the G or metal-rich granule fraction. The concentration of Cd and Zn in each fraction was measured at 0, 7, 14 and 21 days of being fed the flour.The concentration of Cd in the flour affected the concentration of Cd measured in each larval subcellular fraction (p≤0.0001), while the concentration of Zn in the flour only affected the Zn concentration in the S2 and G fractions (p≤0.02). Both Cd and Zn concentrations in mealworms remained relatively constant during the exposure (days 7, 14 and 21) in all three fractions, but the Cd concentrations were much higher than those found in larvae before the exposure (day 0). The concentration of Cd in the flour, however, did not affect the percentage of Cd in the S1 fraction. The contribution of Cd in the G fraction to the total Cd amount was similar (30–40%) in all Cd treatments. The percentage of Zn in all three fractions was not affected by the concentration of Zn in the flour and the relative contributions of each subcellular fraction to the total burden of Zn remained generally constant for both control and treated larvae. In general, larvae sequestered approximately 30% of Cd and Zn in the S1 fraction, which is important for the transport of metals to higher trophic levels in a food web.

Crocodile blood extract induces the apoptosis of lung cancer cells through PTEN activity

Current treatment strategies for lung cancer cause undesirable side‑effects. Integrated medicine with a curative approach has become a common approach to the treatment strategy. Recent studies suggest that American alligator blood is effective in reducing colorectal cancer cell viabilityinvitro, but the mechanism remains unclear. In the present study, we aimed to study the anticancer activity of crocodile blood extracts on lung cancer cell line A549 and investigate the possible mechanisms involved. Invitro studies were utilized to investigate the effects on the cancer cells after incubation with the blood extracts. The active fraction that showed more efficacy in inhibiting cell growth was characterized in the supernatant (S2) from whole blood extracts. High performance liquid chromatography(HPLC) analysis revealed that S2 contained more polar moiety from whole blood. S2 induced DNA fragmentation. Cell cycle arrest in the G1/M phase was demonstrated and mitochondrial membrane permeability was disrupted. An increase in the generation of reactive oxygen species(ROS) and increased activities of caspase-3andcaspase-7 were detected. Furthermore, release of cytochromec, upregulation of expression of Bax, p53, p21, Bid, cleaved forms of the caspase family and PARP along with downregulation of Bcl-2, PCNA, MDM2, caspase‑8, wild types of caspase family proteins and PARP were recorded after treatment with S2 fractions. Moreover, the PI3K/AKT survival pathway was downregulated by S2 fractions in the lung cancer cell line.

Total shale oil inventory from an extended Rock-Eval approach on non-extracted and extracted source rocks from Germany

Rock-Eval pyrolysis data are used for assessments of the quality and maturity of potential hydrocarbon source rock, but is also often applied among shale oil studies to estimate the so-called “total oil” for Oil In Place (OIP) calculations. However, for these approaches Rock-Eval may underestimate total oil due to a “carryover effect”, a bias which is particularly crucial for shale oil investigations.In this study we examined the total oil of marine to lacustrine source rocks from German shale oil formations using data from Rock-Eval combined with solvent extraction. Each source rock was analysed by Rock-Eval both non-extracted and extracted to obtain values for total oil (= [S1wholerock−S1extracted rock]+[S2wholerock−S2extracted rock]). Comparison of these two data sets revealed that for the individual formations in average 17–23% of the S2 fraction is extractable, herein termed “S2 oil”. This “S2 oil” increases the total oil amount compared to the conventional free oil (S1) by average factors of 2.2–3.6. For selected source rocks the molecular composition of the “S2 oil” was obtained by a specific approach: Rock-Eval analysis was terminated after S1-detection at 301°C, samples were retrieved, solvent extracted and analysed by gas chromatography. Depending on the type of organic matter and maturity, the “S2 oil” contains significant amounts of higher boiling hydrocarbons such as n-alkanes but also contributes to the unresolved complex mixture. It is to date unclear, how much of this “S2 oil” should be finally considered as technically mobilized shale oil.

Biological characterization of compounds from Rhinella schneideri poison that act on the complement system.

BackgroundThe skin secretions of toads of the family Bufonidae contain biogenic amines, alkaloids, steroids (bufotoxins), bufodienolides (bufogenin), peptides and proteins. The poison of Rhinella schneideri, formerly classified as Bufo paracnemis, presents components that act on different biological systems, including the complement system. The aim of this study was to isolate and examine the activity of Rhinella schneideri poison (RsP) components on the complement system.MethodsThe components active on the complement system were purified in three chromatographic steps, using a combination of cation-exchange, anion-exchange and gel filtration chromatography. The resulting fractions were analyzed by SDS-PAGE and screened for their activity in the hemolytic assay of the classical/lectin complement pathways. Fractions active on the complement system were also assessed for their ability to generate C3 fragments evaluated by two dimensional immunoelectrophoresis assay, C3a and C5a by neutrophil chemotaxis assay and SC5b-9 complex by ELISA assay.ResultsThe fractionation protocol was able to isolate the component S5 from the RsP, as demonstrated by SDS-PAGE and the RP-FPLC profile. S5 is a protein of about 6000 Da, while S2 presents components of higher molecular mass (40,000 to 50,000 Da). Fractions S2 and S5 attenuated the hemolytic activity of the classical/lectin pathways after preincubation with normal human serum. Both components stimulated complement-dependent neutrophil chemotaxis and the production of C3 fragments, as shown by two-dimensional immunoelectrophoresis. S2 showed a higher capacity to generate the SC5b-9 complex than the other fractions. This action was observed after the exposure of normal human serum to the fractions.ConclusionsThis is the first study to examine the activity of RsP components on the complement system. Fractions S2 and S5 reduced the complement hemolytic activity, stimulated complement-dependent neutrophil chemotaxis and stimulated the production of C3 fragments, indicating that they were able to activate the complement cascade. Furthermore, fraction S2 was also able to generate the SC5b-9 complex. These components may be useful tools for studying dysfunction of the complement cascade.

Traditionally used medicinal plants against uncomplicated urinary tract infections: Are unusual, flavan-4-ol- and derhamnosylmaysin derivatives responsible for the antiadhesive activity of extracts obtained from stigmata of Zea mays L. against uropathogenic E. coli and Benzethonium chloride as frequent contaminant faking potential antibacterial activities?

The dried stigmata from Zea mays L. are used traditionally for the treatment of uncomplicated urinary tract infections. A recent screening has indicated that hydroalcoholic extract of the herbal material inhibits the adhesion of uropathogenic Escherichia coli (UPEC) to T24 bladder cells. For verification of these data EtOH-water (1:1) extracts from 4 different batches of Maydis stigmata were investigated. Within an in vitro adhesion assay (UPEC strain 2980 and human T24 bladder cells) a dose-dependent antiadhesive activity against UPEC was verified (IC50 1040μg/mL). Bioassay guided fractionation of M. stigmata, batch S1, by EtOH-water extraction, followed by chromatography on Sephadex LH20 revealed two active fractions (I and XI). Further purification of fraction I and structure elucidation of the isolated compound revealed the presence of significant amounts of the biocide benzethonium chloride as contaminant. Benzethonium chloride was also identified in subsequent investigations in 2 different batches of M. stigmata. The presence of such nondeclared and illegal contaminants in the herbal raw material market has to be discussed intensively. From benzethonium-free raw material (batch S2) as well as from batch S1 fraction XI was further fractionated by MPLC and preparative HPLC, leading to a still complex subfraction XIG, which was analyzed by UHPLC/+ESI-QTOF-MS analysis. Advanced data processing and species-metabolite relationship database revealed the tentatively existence of the unusual C-glycosidic flavones derhamnosylmaysin (6), 3′-deoxyrhamnosylmaysin (4), 3′-O-methylderhamnosylmaysin (3), apiferol (2) and alternanthin (8) which might be related to the antiadhesive activity of this subfraction against UPEC.