Waters Micromass Quattro

The engineering of photosensitizers (PS) for photodynamic therapy (PDT) with nitric oxide (NO) photodonors (NOPD) is broadening the horizons for new and yet to be fully explored unconventional anticancer treatment modalities that are entirely controlled by light stimuli. In this work, we report a tailored boron-dipyrromethene (BODIPY) derivative that acts as a PS and a NOPD simultaneously upon single photon excitation with highly biocompatible green light. The photogeneration of the two key species for PDT and NOPDT, singlet oxygen (1O2) and NO, has been demonstrated by their direct detection, while the formation of NO is shown not to be dependent on the presence of oxygen. Biological studies carried out using A375 and SKMEL28 cancer cell lines, with the aid of suitable model compounds that are based on the same BODIPY light harvesting core, unambiguously reveal the combined action of 1O2 and NO in inducing amplified cancer cell mortality exclusively under irradiation with visible green light.

Loretta Lazzarato, Elena Gazzano, Marco Blangetti, Aurore Fraix, Federica Sodano, Giulia Maria Picone, Roberta Fruttero, Alberto Gasco, Chiara Riganti, Salvatore Sortino. Antioxidants, 8 (11), 531, 2019.

The aim of this study was to assess the impact of diet and active substances in beetroot juice on the parameters of oxidative stress, inflammation, and muscle damage as well as on the maximum rate of oxygen uptake (VO2max) in elite fencers (10 women, 10 men). Athletes during four weeks realized dietary recommendations (ID) and, after that, diet with freeze-dried beetroot juice supplementation (ID&BEET). At baseline and after each stage, fasting antioxidants, biomarkers of oxidative stress, inflammation, and skeletal muscle damage were measured, and a VO2max test was performed. Only after ID&BEET was a significant increase of VO2max observed, and changes of this parameter were negatively related with changes of serum lactate dehydrogenase (∆LDH) activity, as well as with serum ∆β-carotene and malondialdehyde concentration (∆MDA). Additionally, positive relationships were observed between ∆β-carotene versus changes of the serum concentration of advanced oxidation protein products (∆AOPP), changes of serum glutathione peroxidase activity (∆GPx3) versus both changes of physical activity level and ∆LDH, as well as erythrocyte glutathione peroxidase activity (∆GPx1) versus ∆LDH. To summarize, we showed that long-term beetroot juice supplementation increases lipid peroxidation, and improvement of VO2max after ID&BEET seems to be dependent on LDH activity, as well as on the serum concentration of MDA and β-carotene.

Lucyna Kozłowska, Olga Mizera, Jolanta Gromadzińska, Beata Janasik, Karolina Mikołajewska, Anna Mróz, Wojciech Wąsowicz. Antioxidants, 9 (7), 571, 2020.

Half of human cancers harbor TP53 mutations that render p53 inactive as a tumor suppressor. In these cancers, reactivation of mutant p53 (mutp53) through restoration of wild-type-like function constitutes a valuable anticancer therapeutic strategy. In order to search for mutp53 reactivators, a small library of tryptophanol-derived oxazoloisoindolinones was synthesized and the potential of these compounds as mutp53 reactivators and anticancer agents was investigated in human tumor cells and xenograft mouse models. By analysis of their anti-proliferative effect on a panel of p53-null NCI-H1299 tumor cells ectopically expressing highly prevalent mutp53, the compound SLMP53-2 was selected based on its potential reactivation of multiple structural mutp53. In mutp53-Y220C-expressing hepatocellular carcinoma (HCC) cells, SLMP53-2-induced growth inhibition was mediated by cell cycle arrest, apoptosis, and endoplasmic reticulum stress response. In these cells, SLMP53-2 restored wild-type-like conformation and DNA-binding ability of mutp53-Y220C by enhancing its interaction with the heat shock protein 70 (Hsp70), leading to the reestablishment of p53 transcriptional activity. Additionally, SLMP53-2 displayed synergistic effect with sorafenib, the only approved therapy for advanced HCC. Notably, it exhibited potent antitumor activity in human HCC xenograft mouse models with a favorable toxicological profile. Collectively, SLMP53-2 is a new mutp53-targeting agent with promising antitumor activity, particularly against HCC.

Sara Gomes, Bartolomeo Bosco, Joana B. Loureiro, Helena Ramos, Liliana Raimundo, Joana Soares, Nair Nazareth, Valentina Barcherini, Lucília Domingues, Carla Oliveira, Alessandra Bisio, Silvano Piazza, Matthias R. Bauer, João P. Brás, Maria Inês Almeida, Célia Gomes, Flávio Reis, Alan R. Fersht, Alberto Inga, Maria M. M. Santos, Lucília Saraiva. Cancers, 11 (8), 1151, 2019.

Consumer demand for safe and nutritious fruit juices has led to the development of a number of food processing techniques. To compare the effect of two processing technologies, thermo-sonication (TS) and ultra-high pressure (UHP), on the quality of mango juice, fresh mango juice was treated with TS at 25, 45, 65 and 95 °C for 10 min and UHP at 400 MPa for 10 min. The phenolic profile of mango was also analyzed using the newly developed ultra-performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry (UPLC-Q-TOF-HRMSn) and, based on this result, the effect of TS and UHP on the phenolics of mango juice was evaluated. Both treatments had minimal effects on the oBrix, pH, and titratable acidity of mango juice. The residual activities of three enzymes (polyphenol oxidase, peroxidase, and pectin methylesterase), antioxidant compounds (vitamin C, Total phenolics, mangiferin derivatives, gallotannins, and quercetin derivatives) and antioxidant activity sharply decreased with the increase in the temperature of the TS treatment. Nevertheless, the UHP treatment retained antioxidants and antioxidant activity at a high level. The UHP process is likely superior to TS in bioactive compounds and antioxidant activity preservation. Therefore, the mango juice products obtained by ultra-high-pressure processing might be more beneficial to health.

Abdul Ghani Dars, Kai Hu, Qiudou Liu, Aqleem Abbas, Bijun Xie, Zhida Sun. Foods, 8 (8), 298, 2019.

Protein-energy malnutrition (PEM) is a major health concern in sub-Saharan Africa (SSA). Relying on unexploited and regionally available rich sources of proteins such as insects and sorghum might contribute towards addressing PEM among at-risk populations. Insects are high in nutrients, especially protein, and are abundant in SSA. Sorghum is adapted to the tropical areas of SSA and as such it is an appropriate source of energy compared with temperate cereals like wheat. It is necessary to assess whether cookies fortified with sorghum and termite would be suitable for use in addressing PEM in SSA. Whole grain sorghum meal and termite meal were mixed at a 3:1 ratio (w/w sorghum:termite) to form a sorghum–termite meal blend. Composite cookies were prepared where the sorghum–termite blend partially substituted wheat flour at 20%, 40%, and 60% (sorghum–termite blend:wheat flour (w/w). The functional and nutritional qualities of the cookies were assessed. Compared with the control (100% wheat flour), the cookies fortified with sorghum and termite had about double the quantity of protein, minerals, and amino acids. However, with increased substitution level of the sorghum–termite blend, the spread factor of the cookies decreased. There is a potential to incorporate sorghum and termite in cookies for increased intake of several nutrients by communities that are vulnerable to nutrient deficiencies, especially PEM.

Temitope D. Awobusuyi, Muthulisi Siwela, Kirthee Pillay. Foods, 9 (10), 1427, 2020.

Maize is one of the most interesting dietary sources of hydroxycinnamic acids, widely known for their beneficial health effects, namely antioxidant properties. This work aims to identify hydroxycinnamic acids and their derivatives in broa, a Portuguese traditional ethnic maize bread, and corresponding maize flours. Soluble and insoluble phenolic fractions of diverse maize flours and corresponding broas were prepared and analysed by HPLC-DAD-MS/MS (high-performance liquid chromatography coupled with diode array detector and tandem mass spectrometry). Besides free hydroxycinnamic acids, mainly ferulic and p-coumaric acids, several structural isomers and stereoisomers of insoluble ferulic acid dehydrodimers (n = 18) and trimers (n = 11), were also identified. Hydroxycinnamic acid amides consisting of coumaroyl and feruloyl conjugates (n = 22) were present in both soluble and insoluble fractions of maize flours and breads, in different isomeric forms. A new compound was putatively identified as bis-N,N′-diferuloyl putrescine. Additionally, more complex and insoluble hydroxycinnamic acid amides, derived from ferulic acid dehydrodimers (n = 47) and trimers (n = 18), were also putatively identified for the first time, suggesting that hydroxycinnamic acid amides are also linked to maize cell walls. Since hydroxycinnamic derivatives were not only identified in maize flours, but also in broas, they can contribute to the antioxidant properties and beneficial health effects of maize-based foods.

Andreia Bento-Silva, Noélia Duarte, Elsa Mecha, Maria Belo, Maria Carlota Vaz Patto, Maria do Rosário Bronze. Foods, 9 (10), 1471, 2020.

As one of the most frequently detected pharmaceutical compounds in aquatic environments, carbamazepine (CBZ) has recently been shown to cause acute and chronic toxicity in a variety of non-target aquatic organisms. However, little is known about the ecotoxicological effects it has on the molting and reproduction of crustaceans. The aim of the present work was to evaluate the acute and chronic toxic responses to CBZ in the crustacean Daphnia similis. After acute exposure (4 days), CBZ did not cause lethal toxicity at the tested concentrations. However, CBZ did inhibit the molting and release of chitobiase at concentrations higher than 6.25 μg/L, with 96 h EC50 (median effective concentration) values of 864.38 and 306.17 μg/L, respectively. The results of chronic exposure showed that the mean number of molts, size of the first brood, mean number of offspring per brood, mean number of broods per female, and total offspring per female decreased significantly with increasing CBZ concentrations. Significant effects of CBZ on the molting or fecundity in D. similis were observed even at concentrations as low as 0.03 μg/L. In conclusion, CBZ can cause inhibition of molting, delayed reproduction, and reduced fecundity in D. similis. CBZ toxicity to D. similis depends on the timing and duration of the exposure. Moreover, our results indicated that CBZ would act as an endocrine disrupter in D. similis, as with vertebrates (e.g., fish).

Huihui Chen, Xiaohong Gu, Qingfei Zeng, Zhigang Mao. International Journal of Environmental Research and Public Health, 16 (2), 209, 2019.

Phytoalexins are inducible antimicrobial metabolites in plants, and have been indicated to be important for the rejection of microbial infection. HPLC analysis detected the induced accumulation of three compounds 1–3 in barley (Hordeum vulgare) roots infected by Fusarium culmorum, the causal agent of Fusarium root rot. Compounds 1–3 were identified as cinnamic acid amides of 9-hydroxy-8-oxotryptamine, 8-oxotryptamine, and (1H-indol-3-yl)methylamine, respectively, by spectroscopic analysis. Compounds 1 and 2 had been previously reported from wheat, whereas 3 was an undescribed compound. We named 1–3 as triticamides A–C, respectively, because they were isolated from barley and wheat, which belong to the Triticeae tribe. These compounds showed antimicrobial activities, indicating that triticamides function as phytoalexins in barley. The administration of deuterium-labeled N-cinnamoyl tryptamine (CinTry) to barley roots resulted in the effective incorporation of CinTry into 1 and 2, which suggested that they were synthesized through the oxidation of CinTry. Nine putative tryptamine hydroxycinnamoyl transferase (THT)-encoding genes (HvTHT1–HvTHT9) were identified by database search on the basis of homology to known THT gene sequences from rice. Since HvTHT7 and HvTHT8 had the same sequences except one base, we measured their expression levels in total by RT-qPCR. HvTHT7/8 were markedly upregulated in response to infection by F. culmorum. The HvTHT7 and HvTHT8 enzymes preferred cinnamoyl- and feruloyl-CoAs as acyl donors and tryptamine as an acyl acceptor, and (1H-indol-3-yl)methylamine was also accepted as an acyl acceptor. These findings suggested that HvTHT7/8 are responsible for the induced accumulation of triticamides in barley.

Naoki Ube, Yukinori Yabuta, Takuji Tohnooka, Kotomi Ueno, Shin Taketa, Atsushi Ishihara. International Journal of Molecular Sciences, 20 (22), 5541, 2019.

PIM kinases, a small family of serine/threonine kinases, are important intermediates in the cytokine signaling pathway of inflammatory disease. In this study, we investigated whether the novel PIM kinase inhibitor KMU-470, a derivative of indolin-2-one, inhibits lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 cells. We demonstrated that KMU-470 suppressed the production of nitric oxide and inducible nitric oxide synthases that are induced by LPS in RAW 264.7 cells. Furthermore, KMU-470 inhibited LPS-induced up-regulation of TLR4 and MyD88, as well as the phosphorylation of IκB kinase and NF-κB in RAW 264.7 cells. Additionally, KMU-470 suppressed LPS-induced up-regulation at the transcriptional level of various pro-inflammatory cytokines such as IL-1β, TNF-α, and IL-6. Notably, KMU-470 inhibited LPS-induced up-regulation of a major component of the inflammasome complex, NLRP3, in RAW 264.7 cells. Importantly, PIM-1 siRNA transfection attenuated up-regulation of NLRP3 and pro-IL-1β in LPS-treated RAW 264.7 cells. Taken together, these findings indicate that PIM-1 plays a key role in inflammatory signaling and that KMU-470 is a potential anti-inflammatory agent.

Hye Suk Baek, Hyeon Ji Min, Victor Sukbong Hong, Taeg Kyu Kwon, Jong Wook Park, Jinho Lee, Shin Kim. International Journal of Molecular Sciences, 21 (14), 5138, 2020.

Breast cancer represents the most frequently diagnosed malignancy in women worldwide. Various therapeutics are currently used in order to halt the progression of breast tumor, even though certain side effects may limit the beneficial effects. In recent years, many efforts have been addressed to the usefulness of natural compounds as anticancer agents due to their low toxicity. Resveratrol, a stilbene found in grapes, berries, peanuts and soybeans, has raised a notable interest for its antioxidant, anti-inflammatory, and antitumor properties. Here, we report the design, the synthesis and the characterization of the anticancer activity of a small series of imino N-aryl-substituted compounds that are analogues of resveratrol. In particular, the most active compound, named 3, exhibited anti-tumor activity in diverse types of breast cancer cells through the inhibition of the human topoisomerase II and the induction of apoptotic cell death. Therefore, the abovementioned compound maybe considered as a promising agent in more comprehensive treatments of breast cancer.

Domenico Iacopetta, Rosamaria Lappano, Annaluisa Mariconda, Jessica Ceramella, Maria Stefania Sinicropi, Carmela Saturnino, Marianna Talia, Francesca Cirillo, Fabio Martinelli, Francesco Puoci, Camillo Rosano, Pasquale Longo, Marcello Maggiolini. International Journal of Molecular Sciences, 21 (20), 7797, 2020.

Protein kinases regulate protein phosphorylation, which are involved in fundamental cellular processes such as inflammatory response. In this study, we discovered a novel multi-protein kinase inhibitor, KMU-1170, a derivative of indolin-2-one, and investigated the mechanisms of its inflammation-inhibiting signaling in both THP-1 cells and human osteoarthritic fibroblast-like synoviocytes (FLS). We demonstrated that in THP-1 cells, KMU-1170 inhibited lipopolysaccharide (LPS)-induced upregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and, furthermore, suppressed LPS-induced phosphorylation of transforming growth factor-β-activated kinase 1, JNK, ERK, inhibitor of NF-κB kinase α/β (IKKα/β), and NF-κB p65 as well as nuclear translocation of NF-κB p65. Moreover, KMU-1170 suppressed LPS-induced upregulation of proinflammatory cytokines such as IL-1β, TNF-α, and IL-6, and, notably, inhibited LPS-induced upregulation of the NLRP3 inflammasome in THP-1 cells. Importantly, KMU-1170 attenuated LPS-mediated inflammatory responses in human osteoarthritic FLS, such as the upregulation of IL-1β, TNF-α, IL-6, iNOS, and COX-2 and the phosphorylation of IKKα/β and NF-κB p65. Collectively, these results suggest that KMU-1170 inhibits inflammatory signal transduction and could be developed as a potential anti-inflammatory agent.

Hye Suk Baek, Victor Sukbong Hong, Sang Hyon Kim, Jinho Lee, Shin Kim. International Journal of Molecular Sciences, 22 (3), 1194, 2021.

Renal transplant recipients (RTR) commonly suffer from vitamin B6 deficiency and its functional consequences add to an association with poor long-term outcome. It is unknown whether niacin status is affected in RTR and, if so, whether this affects clinical outcomes, as vitamin B6 is a cofactor in nicotinamide biosynthesis. We compared 24-h urinary excretion of N1-methylnicotinamide (N1-MN) as a biomarker of niacin status in RTR with that in healthy controls, in relation to dietary intake of tryptophan and niacin as well as vitamin B6 status, and investigated whether niacin status is associated with the risk of premature all-cause mortality in RTR. In a prospective cohort of 660 stable RTR with a median follow-up of 5.4 (4.7–6.1) years and 275 healthy kidney donors, 24-h urinary excretion of N1-MN was measured with liquid chromatography-tandem mass spectrometry LC-MS/MS. Dietary intake was assessed by food frequency questionnaires. Prospective associations of N1-MN excretion with mortality were investigated by Cox regression analyses. Median N1-MN excretion was 22.0 (15.8–31.8) μmol/day in RTR, compared to 41.1 (31.6–57.2) μmol/day in healthy kidney donors (p < 0.001). This difference was independent of dietary intake of tryptophan (1059 ± 271 and 1089 ± 308 mg/day; p = 0.19), niacin (17.9 ± 5.2 and 19.2 ± 6.2 mg/day; p < 0.001), plasma vitamin B6 (29.0 (17.5–49.5), and 42.0 (29.8–60.3) nmol/L; p < 0.001), respectively. N1-MN excretion was inversely associated with the risk of all-cause mortality in RTR (HR 0.57; 95% CI 0.45–0.71; p < 0.001), independent of potential confounders. RTR excrete less N1-MN in 24-h urine than healthy controls, and our data suggest that this difference cannot be attributed to lower dietary intake of tryptophan and niacin, nor vitamin B6 status. Importantly, lower 24-h urinary excretion of N1-MN is independently associated with a higher risk of premature all-cause mortality in RTR.

Carolien P.J. Deen, Anna van der Veen, Martijn van Faassen, Isidor Minović, António W. Gomes-Neto, Johanna M. Geleijnse, Karin J. Borgonjen-van den Berg, Ido P. Kema, Stephan J.L. Bakker. Journal of Clinical Medicine, 8 (11), 1948, 2019.

N1-methylnicotinamide (N1-MN) and N1-methyl-2-pyridone-5-carboxamide (2Py) are successive end products of NAD+ catabolism. N1-MN excretion in 24-h urine is the established biomarker of niacin nutritional status, and recently shown to be reduced in renal transplant recipients (RTR). However, it is unclear whether 2Py excretion is increased in this population, and, if so, whether a shift in excretion of N1-MN to 2Py can be attributed to kidney function. Hence, we assessed the 24-h urinary excretion of 2Py and N1-MN in RTR and kidney donors before and after kidney donation, and investigated associations of the urinary ratio of 2Py to N1-MN (2Py/N1-MN) with kidney function, and independent determinants of urinary 2Py/N1-MN in RTR. The urinary excretion of 2Py and N1-MN was measured in a cross-sectional cohort of 660 RTR and 275 healthy kidney donors with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Linear regression analyses were used to investigate associations and determinants of urinary 2Py/N1-MN. Median 2Py excretion was 178.1 (130.3–242.8) μmol/day in RTR, compared to 155.6 (119.6–217.6) μmol/day in kidney donors (p < 0.001). In kidney donors, urinary 2Py/N1-MN increased significantly after kidney donation (4.0 ± 1.4 to 5.2 ± 1.5, respectively; p < 0.001). Smoking, alcohol consumption, diabetes, high-density lipoprotein (HDL), high-sensitivity C-reactive protein (hs-CRP) and estimated glomerular filtration rate (eGFR) were identified as independent determinants of urinary 2Py/N1-MN in RTR. In conclusion, the 24-h urinary excretion of 2Py is higher in RTR than in kidney donors, and urinary 2Py/N1-MN increases after kidney donation. As our data furthermore reveal strong associations of urinary 2Py/N1-MN with kidney function, interpretation of both N1-MN and 2Py excretion may be recommended for assessment of niacin nutritional status in conditions of impaired kidney function.

Carolien P. J. Deen, Anna van der Veen, António W. Gomes-Neto, Johanna M. Geleijnse, Karin J. Borgonjen-van den Berg, M. Rebecca Heiner-Fokkema, Ido P. Kema, Stephan J. L. Bakker. Journal of Clinical Medicine, 9 (2), 437, 2020.