Characterization of the core oligosaccharide and the O-antigen biological repeating unit from Halomonas stevensii lipopolysaccharide: the first case of O-antigen linked to the inner core

A novel core structure among bacterial lipopolysaccharides (LPS) that belong to the genus Halomonas has been characterized. H. stevensii is a moderately halophilic microorganism, as are the majority of the Halomonadaceae. It brought to light the pathogenic potential of this genus. On account of their role in immune system elicitation, elucidation of LPS structure is the mandatory starting point for a deeper understanding of the interaction mechanisms between host and pathogen. In this paper we report the structure of the complete saccharidic portion of the LPS from H. stevensii. In contrast to the finding that the O-antigen is usually covalently linked to the outer core oligosaccharide, we could demonstrate that the O-polysaccharide of H. stevensii is linked to the inner core of an LPS. By means of high-performance anion-exchange chromatography with pulsed amperometric detection we were able to isolate the core decasaccharide as well as a tridecasaccharide constituted by the core region plus one O-repeating unit after alkaline degradation of the LPS. The structure was elucidated by one- and two-dimensional NMR spectroscopy, ESI Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry, and chemical analysis.     

Redox Imbalance and Mitochondrial Release of Apoptogenic Factors at the Forefront of the Antitumor Action of Mango Peel Extract

Today, an improved understanding of cancer cell response to cellular stress has become more necessary. Indeed, targeting the intracellular pro-oxidant/antioxidant balance triggering the tumor commitment to cell demise could represent an advantageous strategy to develop cancer-tailored therapies. In this scenario, the present study shows how the peel extract of mango—a tropical fruit rich in phytochemicals with nutraceutical properties—can affect the cell viability of three colon cancer cell lines (HT29, Caco-2 and HCT116), inducing an imbalance of cellular redox responses. By using hydro-alcoholic mango peel extract (MPE), we observed a consistent decline in thiol group content, which was accompanied by upregulation of MnSOD—a mitochondrial scavenger enzyme that modulates the cellular response against oxidative damage. Such an effect was the consequence of an early production of mitochondrial superoxide anions that appeared after just 30 min of exposure of colon cancer cells to MPE. The effect was accompanied by mitochondrial injury, consisting of the dissipation of mitochondrial membrane potential and a decrease in the level of proteins localized in the mitochondrial membrane—such as voltage-dependent anion-selective channel (VDAC1), mitofilin, and some members of Bcl-2 family proteins (Mcl-1, Bcl-2 and Bcl-X_L)—with the mitochondrial release of apoptogenic factors (cytochrome C and AIF). The analysis of the cytotoxic effects exerted by the different constituents of MPE (gallic acid, mangiferin, citric acid, quinic acid, pentagalloyl glucose, and methyl gallate) allowed us to identify those phytochemicals responsible for the observed anticancer effects, sustaining their future employment as chemopreventive or therapeutic agents.     

Cooperation and game-theoretic cost allocation in stochastic inventory models with continuous review

We study cooperation strategies for companies that continuously review their inventories and face Poisson demand. Our main goal is to analyze stable cost allocations of the joint costs. These are such that any group of companies has lower costs than the individual companies. If such allocations exist they provide an incentive for the companies to cooperate.     

Microwave-assisted extraction and determination of enrofloxacin and danofloxacin photo-transformation products in soil

Here we describe the extraction from soil of the major photo-transformation products (PTPs) of enrofloxacin (ENR) and danofloxacin (DAN), two fluoroquinolones (FQs) widely used in veterinary medicine and of growing environmental concern, because their PTPs have been shown to retain high antibacterial activity. The microwave-assisted extraction (MAE) technique developed previously for determination of FQs, and based on use of an alkaline aqueous solution of Mg(2+) as a complexing agent for the analytes, was applied to agricultural soil samples fortified with different amounts of the PTPs and residues of the parent compounds (53-1000 ng g(-1) for ENR, 24-148 ng g(-1) for DAN). The PTPs, obtained by irradiation of thin layers of the two drugs, were, after extraction, separated and quantified by HPLC-FD. Good recovery (70-130%) and precision (RSDs 1-6% for repeatability and 9-22% for reproducibility) were obtained by use of the overall analytical procedure. The method was applied for the first time to study the in-soil lifecycle of ENR and DAN PTPs, generated in the matrix by irradiation under natural sunlight, at environmentally significant concentrations. Results indicated that soil-adsorbed FQ PTPs are themselves liable to photodegradation and have lifetimes comparable with those of parent compounds.     

A Degenerate Birdcage with Integrated Tx/Rx Switches and Butler Matrix for the Human Limbs at 7 T

The theoretically known degeneracy condition of the band-pass birdcage coil has rarely been exploited in transmit coil designs. We have created an eight-channel degenerate birdcage for the human limbs at 7 T, with dedicated Tx/Rx switches and a Butler matrix. The coil can be split into two half cylinders, as required for its application to patients with limited mobility. The design of the coil, the Butler matrix, and Tx/Rx switches relied on a combination of analytical, circuital, and numerical simulations. The birdcage theory was extended to the degenerate case. The theoretical and practical aspects of the design and construction of the coil are presented. The performance of the coil was demonstrated by simulations, workbench, and scanner measurements. The fully assembled prototype presents good performance in terms of efficiency, B₁ homogeneity, and signal-to-noise ratio, despite the asymmetry introduced by the splittable design. The first in vivo images of the knee are also shown. A novel RF coil design consisting of an eight-channel splittable degenerate birdcage has been developed, and it is now available for 7 T MRI applications of the human lower limbs, including high-resolution imaging of the knee cartilages and of the patellar trabecular structure.     

Enhanced Aryltetralin Lignans Production in Linum Adventi-Tious Root Cultures

Lignans are the main secondary metabolites synthetized by Linum species as plant defense molecules. They are also valuable for human health, in particular, for their potent antiviral and antineoplastic properties. In this study, the adventitious root cultures of three Linum species (L. flavum, L. mucronatum and L. dolomiticum) were developed to produce aryltetralin lignans. The effect of two elicitors, methyl jasmonate and coronatine, on aryltetralin lignans production was also evaluated. The adventitious root cultures from L. dolomiticum were obtained and analyzed for the first time and resulted as the best producer for all the aryltetralins highlighted in this system: Podophyllotoxin, 6-methoxypodophyllotoxin and 6-methoxypodophyllotoxin-7-O-β-glucoside, the last showing a productivity of 92.6 mg/g DW. The two elicitors differently affected the production of the 6-methoxypodophyllotoxin and 6-methoxypodophyllotoxin-7-O-β-glucoside.     

Unexpected detection of 3‐aroylbenzofuran side products in the preparation of 2‐arylbenzofurans: Identification,characterization, and comparison with chalcone's fragmentation patterns using EI/MSn

A gas chromatography‐mass spectrometry study of the intramolecular Wittig reaction revealed, together with the expected 2‐phenylbenzofuran, the formation of an unexpected side product that has not been reported until now. This study reports the identification of the by‐product, ie, the 3‐benzoyl‐2‐phenylbenzofuran, on the base of its mass spectrometric behaviour using a combination of electron ionization, exact mass measurement, multiple stage mass spectrometry, and labelled compounds. This study reports the common fragmentation pathways and discusses possible fragment structures of characteristic ions from a series of 3‐aroyl‐2‐arylbenzofuran derivatives obtained as by‐product under Wittig conditions. Emphasis is laid on the formation and structure investigation of the [M‐H]⁺ and [M‐OH]⁺ ions. Our results showed interesting analogies with the mass spectrometric behaviour of chalcones.     

Untargeted Metabolomics in Forensic Toxicology: A New Approach for the Detection of Fentanyl Intake in Urine Samples

The misuse of fentanyl, and novel synthetic opioids (NSO) in general, has become a public health emergency, especially in the United States. The detection of NSO is often challenged by the limited diagnostic time frame allowed by urine sampling and the wide range of chemically modified analogues, continuously introduced to the recreational drug market. In this study, an untargeted metabolomics approach was developed to obtain a comprehensive “fingerprint” of any anomalous and specific metabolic pattern potentially related to fentanyl exposure. In recent years, in vitro models of drug metabolism have emerged as important tools to overcome the limited access to positive urine samples and uncertainties related to the substances actually taken, the possible combined drug intake, and the ingested dose. In this study, an in vivo experiment was designed by incubating HepG2 cell lines with either fentanyl or common drugs of abuse, creating a cohort of 96 samples. These samples, together with 81 urine samples including negative controls and positive samples obtained from recent users of either fentanyl or “traditional” drugs, were subjected to untargeted analysis using both UHPLC reverse phase and HILIC chromatography combined with QTOF mass spectrometry. Data independent acquisition was performed by SWATH in order to obtain a comprehensive profile of the urinary metabolome. After extensive processing, the resulting datasets were initially subjected to unsupervised exploration by principal component analysis (PCA), yielding clear separation of the fentanyl positive samples with respect to both controls and samples positive to other drugs. The urine datasets were then systematically investigated by supervised classification models based on soft independent modeling by class analogy (SIMCA) algorithms, with the end goal of identifying fentanyl users. A final single-class SIMCA model based on an RP dataset and five PCs yielded 96% sensitivity and 74% specificity. The distinguishable metabolic patterns produced by fentanyl in comparison to other opioids opens up new perspectives in the interpretation of the biological activity of fentanyl.     

Resolution of a Configurationally Stable Hetero[4]helicene

We have developed an efficient chemical resolution of racemic hydroxy substituted dithia-aza[4]helicenes (DTA[4]H) 1(OH) using enantiopure acids as resolving agents. The better diastereomeric separation was achieved on esters prepared with (1S)-(−)-camphanic acid. Subsequent simple manipulations produced highly optically pure (≥ 99% enantiomeric excess) (P) and (M)-1(OH) in good yields. The role of the position where the chiral auxiliary is inserted (cape- vs. bay-zone) and the structure of the enantiopure acid used on successful resolution are discussed.     

Strategies Tackling Viral Replication and Inflammatory Pathways as Early Pharmacological Treatment for SARS-CoV-2 Infection: Any Potential Role for Ketoprofen Lysine Salt?

COVID-19 is an infective disease resulting in widespread respiratory and non-respiratory symptoms prompted by SARS-CoV-2 infection. Interaction between SARS-CoV-2 and host cell receptors prompts activation of pro-inflammatory pathways which are involved in epithelial and endothelial damage mechanisms even after viral clearance. Since inflammation has been recognized as a critical step in COVID-19, anti-inflammatory therapies, including both steroids and non-steroids as well as cytokine inhibitors, have been proposed. Early treatment of COVID-19 has the potential to affect the clinical course of the disease regardless of underlying comorbid conditions. Non-steroidal anti-inflammatory drugs (NSAIDs), which are widely used for symptomatic relief of upper airway infections, became the mainstay of early phase treatment of COVID-19. In this review, we discuss the current evidence for using NSAIDs in early phases of SARS-CoV-2 infection with focus on ketoprofen lysine salt based on its pharmacodynamic and pharmacokinetic features.