Analysis of bitter limonoids in citrus juices by atmospheric pressure chemical ionization and electrospray ionization liquid chromatography-mass spectrometry

Improved analytical techniques for bitter limonoids in citrus and citrus juices can expedite the evaluation of freeze-induced citrus damage for citrus growers and juice quality for citrus juice producers. Microbore normal-phase and reverse-phase chromatography coupled to a mass spectrometer operating in a positive ion atmospheric pressure chemical ionization and electrospray ionization modes were found to be rapid, selective, and sensitive methods for the analysis of the bitter limonoids limonin and nomilin in citrus juices. Analysis was performed on a chloroform extract of citrus juice to which an internal standard was added. The methods are capable of detecting citrus limonoids in citrus juice in the 60-200 picogram range and quantifying citrus juice limonoids in concentrations as low as 120 picograms. An accurate "total limonoid bitterness" in citrus juice, as represented by the combined occurrence of limonin and nomilin, is easily determined by these methods.     

ADVERSE EFFECTS OF HUMIC SUBSTANCES FROM DIFFERENT ORIGIN ON LUPIN AS RELATED TO IRON SOURCES

Humic substances improve the efficiency of different iron (Fe) sources overcoming Fe deficiency chlorosis of plants. However, applied at high rates, they can promote negative effects on plants. The main objective of this work was to study the potential adverse effect of three humic acids from different origin when they were applied with two effective Fe sources for plants: Fe- ethylenediaminedihydroxyphenylacetic acid (EDDHA) and Vivianite. To this end, an experiment with lupin (Lupinus albus L.) was performed involving two factors: (i) Fe source, and (ii) humic substances from three different origin (composted cork, leonardite, and compost obtained from a mixture of olive husk with cotton gin trash) applied at 0, 0.1, and 0.5 g organic carbon (C) kg^?1 of growing media. At the rates used, humic substances promoted adverse effects on plant development, chlorophyll meter readings, and Fe content in lupin grown in calcareous media. Overall, the effect on dry matter and Fe content in plants was more relevant when Fe was supplied with Vivianite, the effect on chlorophyll meter readings being more significant when Fe was applied as Fe-EDDHA. Differences were also observed depending on the source of humic substances, those from leonardite promoting the greatest decrease in dry matter in roots and shoots. These humic substances possessed the highest values of spectroscopy index for aromaticity (A₂₅₄ ). On the other hand, the application of humic substances from olive husk compost, which exhibited the lower aromaticity index, resulted in the smallest decrease in dry matter production and chlorophyll meter readings. Dry matter in roots decreased logarithmically with increased values of the estimates of the amounts of aromatic compounds accumulated in the growing media (R² = 0.92; P < 0.01) with Vivianite as Fe source. Thus, the effects decreasing dry matter production, particularly in roots, and chlorophyll meter readings can be ascribed at least partially to the presence of phytotoxic aromatic compounds in humic substances.     

Nomenclature and genetic relationships of apples and pears from Terceira Island

Heritage apple (Malus domestica Borkh. hybrids) and pear (Pyrus communis L. hybrid) trees grow in villages throughout Terceira Island, Azores, Portugal. Some of these pears have different names but similar morphology. The objective of this study was to determine synonymy, homology, and phylogeny of apples and pears from Terceira and to examine potential relationships of the island pears with standard apples and pears of Portuguese or American descent. Nine apple microsatellite markers were used to determine genetic relationships. Distance- and parsimony-based cluster analysis grouped these genotypes into separate apple and pear clades. The Terceira apples were divided into two clades: the maçā and the reineta-reinette. Among the 17 heritage apple genotypes, seven unique accessions were identified and four groups of synonyms, or possibly clones, were detected including: ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares; ‘Reineta Castanha’ and ‘Reineta Verde Miuda’; ‘Maçā Pêra,’ ‘Maçā Calhau’, ‘Pêro Branco’ from Salga and from Terra-Chā and ‘Maçā Marmelo’; and the five genotypes ‘Maçā Sao Joao’, ‘Malápio Rosa’, ‘Maçā Gaspar’, ‘Maçā Branca’ and ‘Maçā Pato’. In addition, two homonyms were detected. ‘Pêro Vermelho’ from Terra Chā was a separate genotype from a tree from Doze Ribeiras of the same name, but Pêro Branco from Terra Chā appears to be a clone that can be distinguished by an additional allele at CH1F07a from a tree with that name from Salga. One pair of apple clones, ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares appear to be derived from an unreduced gamete of ‘Golden Delicious.’ Another apple genotype ‘Maçā Acida’ could be a sibling of the ‘Maçā Pêra’ clonal group. Other tested standard apples from the US genebank were unrelated to Terceira genotypes. Of the seven heritage pears, five unique genotypes and one pair of synonyms were detected. ‘Pêra Papo Pintassilgo’ from Raminho and ‘Pêra Vermelha’ from the nursery of Serviço de Desenvolvimento Agario da Terceira (SDAT) were synonyms. ‘Passans du Portugal’ was related to ‘Pêra Cabaca’ but other standard pears from the US genebank were unrelated to Terceira genotypes. Future studies will include additional apple and pear cultivars from other Islands of the Azores and continental Portugal, and wild Asian species to further explore genetic relationships.     

Culture system and long-term storage of culture media in the in vitro production of bovine embryos

The optimum culture system for in vitro matured and fertilised oocytes still remains to be clarified. Culture media (CM) for mammalian embryos are routinely prepared fresh for use and preserved under refrigeration during one or two weeks. The purposes of this work were (1) to compare the efficiency of a synthetic oviduct fluid (SOF) with two different bovine serum albumin (BSA) concentrations (3 and 8 g/L) for the in vitro production of bovine blastocysts, (2) to test the effect of timing on adding fetal calf serum (FCS) to the SOF, and (3) to evaluate the effects on bovine embryo development of freezing and lyophilisation as procedures for preserving the SOF. Supplementation of SOF with 3 g/L BSA increased Day-7 blastocyst expansion rates (18.3 ± 1.6 vs. 14.4 ± 0.7; P < 0.05), although no differences in hatching rates were found. Addition of FCS to SOFaa (SOF with amino acids) medium supplemented with sodium citrate (SOFaaci) at 48 and at 72 h post-insemination (PI) allowed obtaining higher Day-6 embryo development rates than when FCS was added at 18 or 96 h PI (Day-6 morulae + blastocyst rate: 30.0 ± 1.1, 40.8 ± 1.1, 43.9 ± 2.3 and 39.3 ± 0.5 for FCS addition at 18, 48, 72 and 96 h, respectively). Hatching rates were significantly improved when serum was added at 72 h PI. Finally, both refrigeration and lyophilisation appeared as useful cryopreservation procedures for SOFaaci, although a significant loss of its ability to support embryo development, compared to the control fresh culture medium, was observed.     

Evaluation of alternative sanitizers to chlorine disinfection for reducing foodborne pathogens in fresh-cut apple

The risk of undesirable by-products from chlorine disinfection in fresh-cut industries, together with its limited efficacy, has led to a search for alternative agents. The aim of this study was to test several alternative putative antimicrobial substances to reduce Escherichia coli O157:H7, Salmonella spp. and Listeria spp. populations on fresh-cut apple. Carvacrol, vanillin, peroxyacetic acid, hydrogen peroxide, N-acetyl-l-cysteine and Citrox were selected for their results in in vitro assays against E. coli O157:H7 and Listeria spp., to be tested on fresh-cut apple plugs. Apple flesh was inoculated by dipping in a suspension of a mix of the studied pathogens at 10⁶ cfu mL^?1, and then treated with the antimicrobial substances. All treatments were compared to deionized water and a standard sodium hypochlorite treatment (SH, 100 mg L^?1, pH 6.5). Pathogen population on apple plugs was monitored for up to 6 days at 10 °C. Bacterial reductions obtained by peroxyacetic acid (80 and 120 mg L^?1), vanillin (12 g L^?1), hydrogen peroxide (5, 10, 20 mL L^?1) and N-acetyl-l-cysteine (5 and 10 g L^?1) were similar or higher than reduction obtained by SH. In addition, bacterial populations were maintained at low levels throughout storage. No cells of any of the pathogens were detected in the peroxyacetic acid, hydrogen peroxide, Citrox and SH washing solutions after apple treatment. Peroxyacetic acid, hydrogen peroxide and N-acetyl-l-cysteine could be potential disinfectants for the fresh-cut industry as an alternative to chlorine disinfection. However, their effect on sensory quality and effectiveness under commercial processing conditions should be evaluated.     

Mitotic Golgi partitioning is driven by the membrane-fissioning protein CtBP3/BARS

Organelle inheritance is an essential feature of all eukaryotic cells. As with other organelles, the Golgi complex partitions between daughter cells through the fission of its membranes into numerous tubulovesicular fragments. We found that the protein CtBP3/BARS (BARS) was responsible for driving the fission of Golgi membranes during mitosis in vivo. Moreover, by in vitro analysis, we identified two stages of this Golgi fragmentation process: disassembly of the Golgi stacks into a tubular network, and BARS-dependent fission of these tubules. Finally, this BARS-induced fission of Golgi membranes controlled the G2-to-prophase transition of the cell cycle, and hence cell division.     

Effect of Fucoidan on the Mitochondrial Membrane Potential (ΔΨm) of Leukocytes from Patients with Active COVID-19 and Subjects That Recovered from SARS-CoV-2 Infection

Fucoidan is a polysaccharide obtained from marine brown algae, with anti-inflammatory, anti-viral, and immune-enhancing properties, thus, fucoidan may be used as an alternative treatment (complementary to prescribed medical therapy) for COVID-19 recovery. This work aimed to determine the ex-vivo effects of treatment with fucoidan (20 µg/mL) on mitochondrial membrane potential (ΔΨm, using a cationic cyanine dye, 3,3′-dihexyloxacarbocyanine iodide (DiOC₆(3)) on human peripheral blood mononuclear cells (HPBMC) isolated from healthy control (HC) subjects, COVID-19 patients (C-19), and subjects that recently recovered from COVID-19 (R1, 40 ± 13 days after infection). In addition, ex-vivo treatment with fucoidan (20 and 50 µg/mL) was evaluated on ΔΨm loss induced by carbonyl cyanide 3-chlorophenylhydrazone (CCCP, 150 µM) in HPBMC isolated from healthy subjects (H) and recovered subjects at 11 months post-COVID-19 (R2, 335 ± 20 days after infection). Data indicate that SARS-CoV-2 infection induces HPBMC loss of ΔΨm, even 11 months after infection, however, fucoidan promotes recovery of ΔΨm in PBMCs from COVID-19 recovered subjects. Therefore, fucoidan may be a potential treatment to diminish long-term sequelae from COVID-19, using mitochondria as a therapeutic target for the recovery of cellular homeostasis.     

Six Edible Wild Fruits as Potential Antioxidant Additives or Nutritional Supplements

Total antioxidant activity of six non-cultivated but traditionally collected fruits from the south of Europe was assessed by measuring their ability to reduce the hydroxyl radical ( OH^· ) \left( {{\hbox{O}}{{\hbox{H}}^\bullet }} \right) and hydrogen peroxide (H₂O₂), and their Trolox equivalent antioxidant capacity (TEAC). This antioxidant activity was compared with that shown by the synthetic antioxidants BHA (E-320), BHT (E-321) and propyl gallate (E-310). Total phenolics, ascorbic acid and the carotenoid content of the fruits were also analyzed. All fruits showed a high ability to scavenge the OH^· {\hbox{O}}{{\hbox{H}}^\bullet } radical, ranging from 60.61% to 81.04% inhibition for Rosa canina and Crataegus monogyna, respectively. The H₂O₂ scavenging capacity and the TEAC value varied widely, ranging between 3.63% and 87.26% inhibition of H₂O₂ and between 0.47 and 416.64 mM trolox g⁻¹ FW for Sorbus domestica and Rosa canina, respectively. The antioxidant activity of fruits was higher than that of the synthetic additives analyzed, except in the TEAC assay. The phenolic and carotenoid content of R. canina was much higher than that of the other fruits analyzed and its ascorbic acid concentration was also high, reflecting its higher efficacy towards ABTS ^·- {\hbox{ABT}}{{\hbox{S}}^{ \bullet - }} (TEAC assay) and H₂O₂ species_. In spite of these associations, the correlation coefficients between total antioxidant activity and the antioxidant compounds analyzed were not very significant; only phenolics and carotenoids showed a marginal correlation with the TEAC assay. The results support the possible use of R. canina as natural antioxidant to replace the synthetic additives, as well as their use in the production of functional foods with a high antioxidant activity.     

Chemical composition and pasting properties of einkorn (Triticum monococcum L. subsp. monococcum) whole meal flour

Einkorn (Triticum monococcum L. subsp. monococcum) is an ancient wheat appreciated for its high proteins, carotenoids and tocols content. To better understand its potential for human consumption and food preparation, composition and pasting properties of 65 einkorn accessions, of different geographical origin but cultivated at S. Angelo Lodigiano (Italy), were evaluated. Eight Triticum turgidum and seven Triticum aestivum cultivars, belonging to different subspecies, were assessed as controls.On average, the einkorn samples had light seeds (25.0 mg/kernel), extra-soft texture (238.3 g), high protein content (18.2%), high ash content (2.35%), low SDS sedimentation volume (25.6 ml), high carotenoids (8.36 μg/g) and high yellow pigments (8.46 μg/g). The pasting parameters studied were peak viscosity (average: 2426 cP), breakdown (765 cP), final viscosity (2788 cP) and setback (1126 cP). Total amylose (25.7%) and total starch (65.5%) were also measured. Significant differences from the controls and a broad variation for all the traits analysed were observed. Einkorns from diverse geographical areas showed different mean values for all characters, apart from ash content.The good pasting properties, coupled with high proteins and carotenoids content, suggest that einkorn is particularly suited for the production of baby and specialty foods.     

Chemical markers for the evaluation of raw material hygienic quality in egg products

The aim of this research was to study uracil and lactic and acetic acids as chemical markers for hygienic quality evaluation of raw material in liquid pasteurized egg products. Uracil, absent in sound whole eggs, was formed in raw and pasteurized egg products as a consequence of high microbial contamination (>10(6) cfu/g) after a sufficient lag time, remaining stable at 4 degrees C but disappearing after 7 days of storage at 25 degrees C. Both lactic and acetic acids, starting from initial values of 1-7 mg/kg dry matter, presented trends similar to that of uracil; however, acetic acid never decreased during the storage of raw egg products. With few exceptions, all three metabolites were produced by Enterobacter cloacae, Escherichia coli, Morganella morganii, Serratia liquefaciens, Aeromonas hydrophyla, Pseudomonas fluorescens, Enterococcus avium, and Enterococcus faecalis, separately inoculated in whole egg samples. Uracil seems to be the most sensible marker, with a suggested limit corresponding to the detectable level.