Bioconjugation of carbohydrates to gelatin sponges promoting 3D cell cultures

Gelatin sponges are widely employed as hemostatic agents, and are gaining increasing interest as 3D scaffolds for tissue engineering. To broaden their possible application in the field of tissue engineering, a straightforward synthetic protocol able to anchor the disaccharides, maltose and lactose, for specific cell interactions was developed. A high conjugation yield was confirmed by 1H-NMR and FT-IR spectroscopy, and the morphology of the resulting decorated sponges was characterized by SEM. After the crosslinking reaction, the sponges preserve their porous structure as ascertained by SEM. Finally, HepG2 cells cultured on the decorated gelatin sponges show high viability and significant differences in the cellular morphology as a function of the conjugated disaccharide. More spherical morphologies are observed when cultured on maltose-conjugated gelatin sponges, while a more flattened aspect is discerned when cultured onto lactose-conjugated gelatin sponges. Considering the increasing interest in small-sized carbohydrates as signaling cues on biomaterial surfaces, systematic studies on how small carbohydrates might influence cell adhesion and differentiation processes could take advantage of the described protocol

Interaction of Cisplatin with a CCHC Zinc Finger Motif

The interaction between cisplatin and an 18-residue CCHC zinc finger motif derived from a retroviral nucleocapsid protein (PyrZf18) has been studied using UV–visible, CD and 1H NMR spectroscopies and ESI-MS spectrometry. Cisplatin irreversibly blocks the cysteine zinc binding groups in the free peptide and is able to slowly eject zinc from the zinc–peptide complex. The observed end product of the reaction with cisplatin is a complex in which only one ammonia molecule is coordinated to platinum. After an initial binding with two cysteine residues and the formation of the (PyrZf18)– platinum–(NH3)2 complex, a release of one ammonia molecule occurs because of trans-labilization, and the third cysteine is coordinated, leading to a mixture of isomers and/or conformers of the (PyrZf18)–platinum–NH3 complex. The results are discussed with respect to the potential antiretroviral activity of platinum(II) compounds and to the possible interaction of cisplatin with the cellular nucleic acid binding proteins

Electrokinetic remediation of metal-polluted marine sediments: experimental investigation for plant design

This paper presents the results of an extensive set of laboratory experiments performed to design a demonstrative electrokinetic plant for extracting heavy metals from marine sediments dredged from the Livorno marine harbour. The investigated sediments displayed a high salinity, a high acid neutralization capacity, a low electrical resistivity (0.5 Ωm), a high alkalinity (pH ≈ 8) and a large fraction of fine particles. The target metals were Cd, Cr, Cu, Ni, Pb and Zn at relatively weak and inhomogeneous concentrations with high non-mobile fractions. After an accurate characterization, several screening and full electrokinetic tests were performed using cells of two different sizes, several conditioning agents (HNO3, HCl, H2SO4, citric acid, oxalic acid, ascorbic acid, EDTA), different applied current intensities and durations. The tests highlighted the need for long treatment times in order to obtain a significant pH reduction, with some appreciable metal removal being attained only after several weeks. The best results were obtained with strong acids used as the conditioning agents, with significant specific effects of each acid, including pronounced resistivity increase (from 0.5 up to 10 Ωm) and a high electroosmotic flow (EOF) with H2SO4, or a reversed EOF (electroendosmosis), and minor resistivity changes with HNO3. The use of the obtained data to design a demonstrative plant is also presented in the paper, with considerations on operating parameters such as energy and reagent consumption, characteristics of plant components and required safety measures. buffer capacity, conditioning agent

Phytochemical profile, antioxidant and antidiabetic activities of Adansonia digitata L. (baobab) from Mali, as source of health-promoting compounds

Background: Adansonia digitata L. (Malvaceae), also known as baobab, is a tree attracting recent interest especially due to the high nutritional value of the fruit pulp. However, few studies are reported on the secondary metabolite content, showing high variability due to geographic region. Methods: In this study, the chemical profiles of Malian commercial baobab fruits and leaves, focused on phenolic content, were investigated by HPLC coupled with a photodiode array (PDA)/UV and an electrospray ionization (ESI) mass spectrometer (MS) and gas chromatography (GC)/MS. In addition, the extracts of fruit pulps obtained from three different markets (Fruits 1, 2, and 3) were evaluated for their total phenolic content (TPC), antioxidant activity and α-glucosidase inhibition. Results: Baobab fruit pulps were found to be rich in procyanidins and flavonol glycosides, with tiliroside as the major constituent. The baobab leaves showed a similar profile respect to the fruits, but with more detected phenolics. All fruit pulp extracts exerted antioxidant activity (highest for Fruit 3) and higher α-glucosidase inhibition than acarbose used as standard. Conclusions: This study confirmed the variability of baobab with different origins and indicated Malian species baobab as promising source of health-promoting substances

Are the Follicular Fluid Characteristics of Recovered Coronavirus Disease 2019 Patients Different From Those of Vaccinated Women Approaching in vitro Fertilization?

The aim of this pilot study is to evaluate if SARS-CoV-2 infection or vaccination against SARS-CoV-2 infection induce observable metabolic effects in follicular fluid of women who are following in vitro fertilization (IVF) treatments. The possible impact of coronavirus disease 2019 (COVID-19) on fertility and IVF outcome is considered. We have selected for this study: six women vaccinated against SARS-CoV-2 infection, five recovered COVID-19 patients, and we used nine healthy women as the control group. At the time of oocytes retrieval from participants in the study, follicular fluids were collected and metabolomic analysis was performed by 1H NMR spectroscopy in combination with multivariate analysis to interpret the spectral data. The search for antibody positivity in the follicular fluid aspirates was also carried out, together with the western blotting analysis of some inflammatory proteins, interleukin-6, tumor necrosis factor α (TNFα), and the free radical scavenger superoxide dismutase 2. Higher levels of Ala and Pro together with lower levels of lipids and trimethylamine N-oxide (TMAO) were found in follicular fluids (FFs) of vaccinated women while lower levels of many metabolites were detected in FFs of recovered COVID patients. Expression level of TNF-α was significantly lower both in recovered COVID-19 patients and vaccinated women in comparison to healthy controls

Induction of OCT2 contributes to regulate the gene expression program in human neutrophils activated via TLR8

The transcription factors (TFs) that regulate inducible genes in activated neutrophils are not yet completely characterized. Herein, we show that the genomic distribution of the histone modification H3K27Ac, as well as PU.1 and C/EBP beta, two myeloid-lineage-determining TFs (LDTFs), significantly changes in human neutrophils treated with R848, a ligand of Toll-like receptor 8 (TLR8). Interestingly, differentially acetylated and LDTF-marked regions reveal an over-representation of OCT-binding motifs that are selectively bound by OCT2/POU2F2. Analysis of OCT2 genomic distribution in primary neutrophils and of OCT2-depletion in HL-60-differentiated neutrophils proves the requirement for OCT2 in contributing to promote, along with nuclear factor kappa B (NF-kappa B) and activator protein 1 (AP-1), the TLR8-induced gene expression program in neutrophils. Altogether, our data demonstrate that neutrophils, upon activation via TLR8, profoundly reprogram their chromatin status, ultimately displaying cell-specific, prolonged transcriptome changes. Data also show an unexpected role for OCT2 in amplifying the transcriptional response to TLR8-mediated activation

New insights on the functional role of URG7 in the cellular response to ER stress.

BACKGROUND INFORMATION: Up-regulated Gene clone 7 (URG7) is an ER resident protein, whose expression is upregulated in the presence of hepatitis B virus X antigen (HBxAg) during HBV infection. In virus-infected hepatocytes, URG7 shows an anti-apoptotic activity due to the PI3K/AKT signaling activation, does not seem to have tumorigenic properties, but it appears to promote the development and progression of fibrosis. However, the molecular mechanisms underlying URG7 activity remain largely unknown. RESULTS: To shed light on URG7 activity, we first analyzed its interactome in HepG2 transfected cells: this analysis suggests that URG7 could have a role in affecting protein synthesis, folding and promoting proteins degradation. Moreover, keeping into account its subcellular localization in the ER and that several viral infections give rise to ER stress, a panel of experiments was performed to evaluate a putative role of URG7 in ER stress. Our main results demonstrate that in ER stressed cells URG7 is able to modulate the expression of Unfolded Protein Response (UPR) markers toward survival outcomes, upregulating GRP78 protein and downregulating the pro-apoptotic protein CHOP. Furthermore, URG7 reduces the ER stress by decreasing the amount of unfolded proteins, by increasing both the total protein ubiquitination and the AKT activation and reducing caspase 3 activation. CONCLUSIONS: All together these data suggest that URG7 plays a pivotal role as a reliever of ER stress-induced apoptosis. SIGNIFICANCE: This is the first characterization of URG7 activity under ER stress conditions. The results presented here will help to hypothesize new strategies to counteract the antiapoptotic activity of URG7 in the context of the viral infection. This article is protected by copyright. All rights reserved