Starches from various botanic origins (maize, quinoa, wheat, potato and rice) were studied. The thermal and pasting properties and their connection with enzyme digestibility were evaluated. Various hydrothermal treatments were applied, taking the starch physical parameters into account, in order to obtain partial and total gelatinisation of the starch structure and determine its influence on enzymatic action. Onset and pasting temperatures of the gelatinisation and pasting processes, respectively, followed the same order in the cereal starches (rice > maize > wheat > quinoa). These results were accompanied by an opposite trend in the percentage of raw starch hydrolysis, with quinoa reaching a level more than 2-fold higher than that of raw maize starch in in vitro digestion kinetics. Other technological parameters, such as high peak viscosity or low breakdown, also reflected modifications in the quinoa starch structure which were related to improved digestibility. However, starch from potato, the only tuber, displayed different characteristics from those of cereal starch, showing greater resistance to digestion. When the starches were pretreated, digestibility increased in all of them compared to their raw counterparts, with the pretreated quinoa and wheat starches showing greater susceptibility to modification of their structure. Although the hydrothermally pretreated maize and rice starches reached about 75% of the hydrolysis index of the corresponding gelatinised starches, raw quinoa had a similar hydrolysis index and quinoa obtained a higher value for total starch hydrolysed. Thus, quinoa starch could be potentially beneficial in the design of more digestible formulations for patients with metabolic disorders such as glycogen storage disease, among others. | This work was financially supported by grants QuiSalhis-Food (AGL2016-75687-C2-1-R) from the Ministry of Science, Innovation and Universities (MICIU) and CYTED, LA ValSe-Food (119RT0S67). The contract given to R. Selma-Gracia as part of LINCE (PROMETEO/2017/189) by the Generalitat Valenciana is gratefully acknowledged. | Peer reviewed

Pompe disease, also known as glycogen storage disease type II, is caused by the lack or deficiency of a single enzyme, lysosomal acid alpha-glucosidase, leading to severe cardiac and skeletal muscle myopathy due to progressive accumulation of glycogen. The discovery that acid alpha-glucosidase resides in the lysosome gave rise to the concept of lysosomal storage diseases, and Pompe disease became the first among many monogenic diseases caused by loss of lysosomal enzyme activities. The only disease-specific treatment available for Pompe disease patients is enzyme replacement therapy (ERT) which aims to halt the natural course of the illness. Both the success and limitations of ERT provided novel insights in the pathophysiology of the disease and motivated the scientific community to develop the next generation of therapies that have already progressed to the clinic.

Mangoes tend losses of fruit quality due to ripening and deteriorating quickly during storage at room temperature. In this study, the effect of postharvest calcium treatments with two different concentrations on storability and disease resistance was investigated in pre‐bagging mango during storage at room temperature. The results showed that the calcium treatments delayed fruit ripening and decreased fruit sweetness in pre‐bagging mango fruit. Both 1% and 2% of calcium treatment could effectively maintain lower level of hydrogen peroxide (H₂O₂) via higher activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) during the late stage of storage, which might be beneficial to delaying fruit ripening and enhancing disease resistance in mango fruit. Moreover, a significant decrease in the disease index in calcium‐treated fruit might be attributed to the increases in activities of defense‐related enzymes and the accumulations of disease resistance‐related substances. PRACTICAL APPLICATIONS: “Keitt” mango is susceptible to fruit fly and deteriorates quickly during room temperature storage. Therefore, finding a complementary postharvest approach to improve fruit storability and quality of mango is beneficial for producers and traders. Calcium is thought to be one of the most important mineral elements associated with fruit quality and storability. Our results suggested that postharvest calcium treatment might be a useful method to improve storability and enhance disease resistance in pre‐bagging mango fruit during storage.

Loss-of-function of the glucose-6-phosphate transporter is caused by biallelic mutations in SLC37A4 and leads to glycogen storage disease Ib. Here we describe a second disease caused by a single dominant mutation in the same gene. The mutation abolishes the ER retention signal of the transporter and generates a weak Golgi retention signal. Intracellular mislocalization of the transporter leads to a congenital disorder of glycosylation instead of glycogen storage disease.

Patients and Methods: A retrospective chart review of 32 GSD- I patients, followed at the American University of Beirut Medical Center, between 2007 and 2018 was conducted. Diagnosis was confirmed by enzymatic and/or genetic studies. Clinical presentation, growth, and kidney outcome were assessed. All patients were evaluated for body mass index, blood parameters of metabolic control including uric acid, alanine, lactic acid, and triglycerides in blood. Kidney evaluation included creatinine clearance, microalbuminuria, citraturia, and calciuria as well as urine microalbumin/creatinine ratio. Results: Almost one third of GSD-I patients developed microalbuminuria. This was detected below 7 months of age in 36% of patients who required early treatment with ACEI with significant reduction in albuminuria. Kidney stones were present in 6% and were associated with hypercalciuria and hypocitraturia. Poor metabolic control reflected by hyperuricemia, lactic acidosis, and hyperalaninemia were noted only in patients who developed microalbuminuria. Conclusion: Glomerular injury may appear in early infancy in poorly controlled patients. Adequate metabolic control and ACEI therapy may improve kidney outcome in GSD I patients. Plasma alanine appears to be a promising and reliable marker reflecting metabolic control in GSD-I patients. © Copyright © 2020 Aoun, Sanjad, Degheili, Barhoumi, Bassyouni and Karam.

UV-B radiation (280-320 nm) is known to elicit multiple protective responses in plants, including antioxidant defense system and synthesis of phenolic compounds. The objective of this work was to investigate the potential of pre-storage treatment with UV-B light to enhance disease resistance and health-beneficial phyto-compounds in carrots. Fresh carrots (LSQUOSun255RSQUO) were treated with UV-B (0.0-14.0 kJ m(‑2)) and stored at 4°C. The disease resistance of treated carrots was determined by challenge-inoculation with Botrytis cinerea. The titers of phyto-compounds of significance in disease resistance; the GC-MS secondary metabolite profiles; and the physiological and quality characteristics of treated carrots were monitored at regular intervals during six weeks of storage. The UV-B dose-disease resistance response relationship was bi-phasic, and the hormetic dose of UV-B for the induction of disease resistance was 7.0 kJ m(‑2), where the disease inhibition was about 70%. The disease resistance strongly related to the accumulation of phytoalexin 6-MM. UV-B treatment also greatly enhanced the accumulation of myristicin, with its titer increasing with increasing UV-B dose. However, the levels of polyacetylenes were not affected significantly by the treatment. Furthermore, the treatment enhanced the levels of phytochemicals, including phenylpropanoids, terpenoids, and iso-coumarins. The initial respiration rate and electrolyte leakage of treated carrots were higher immediately after the exposure to UV-B, which increased with increasing dose; but they gradually decreased to steady state levels comparable to the control. In addition, the weight loss and antioxidant capacity of the treated carrots were not affected significantly during storage. The results suggest a strong potential for pre-storage treatment with UV-B to preserve fresh carrots by controlling diseases, maintaining quality, and enhancing the levels of plant protective and health-beneficial phyto-compounds.

This thesis aimed at describing the pathology of a novel progressive neurologic disease in dogs of the Lagotto Romagnolo (LR) breed, at testing the hypothesis of altered autophagy in affected dogs, derived from genetic investigations, and at testing the hypothesis of a lysosomal storage disease affecting the dogs, derived from the histopathologic changes. Autophagy is a degradative cellular process, responsible for the turnover of damaged organelles and clearance of protein aggregates, that is disturbed in several neurodegenerative diseases in man. Histologically, altered autophagy commonly leads to neuronal proteinaceous inclusions as well as spheroid formation. Occasionally, neuronal vacuoles have been linked to altered autophagy. Cytoplasmic vacuolation is histologically more typical for lysosomal storage diseases, a group of inherited diseases characterized by defective lysosomal degradation, which often affect the nervous system. Compensatory rerouting of cargo during defective autophagy and lysosomal degradation may affect the content and release of extracellular vesicles (EVs) from cells. Post mortem examination of 13 affected LRs confirmed a distinct histopathological phenotype, consisting of intracellular vacuole storage in neurons, axonal damage and neuronal loss. Secretory epithelial cells and selected mesenchymal cells were also affected by vacuolation. In order to link the histopathologic findings to altered autophagy, affected organs and age- as well as breed matched control tissue, were immunohistochemically stained with markers for autophagosomal, lysosomal and endosomal membranous antigens, autophagy receptors and autophagic cargo. Electron microscopy and immunoelectron microscopy with the same membranous markers were used in order to obtain more specific results regarding the subcellular morphological changes. Autophagy was functionally monitored by the LC3 I/II conversion assay and by quantifying autophagosomes with immunofluorescence in cultured fibroblasts from affected dogs in comparison to control. The screening for a lysosomal storage disease included urine analysis to detect excessive oligosaccharide or glucosaminoglycan excretion. Histochemical staining and electron microscopy were used to examine the affected tissues for lysosomally stored material. In addition, the activity of three lysosomal enzymes were measured in fibroblasts from affected dogs and controls, as well as in culture medium, in order to identify alterations in lysosomal function. The size, amount and cargo of EVs released from fibroblasts of affected dogs were compared to those released from control cells, in order to detect changes in EV release or content that could support or contradict either hypothesis. Nano-particle tracking analysis of EVs, as well as mass spectrometry and functional enrichment analysis of the EV proteomes were performed. Both autophagosomal markers and autophagic cargo accumulated within the lesions of the nervous system in affected dogs. The stored vacuoles in affected tissues represented hybrid organelles of the autophagic and endolysosomal pathways, displaying membranous markers of both pathways. Fibroblasts showed changes in basal, but not starvation-induced autophagy, as both basal LC3II levels and basal number of LC3 positive spots were significantly elevated. In addition, the basal EV release was significantly increased and the basal EV proteome enriched in substrates of basal autophagy in the cells of affected dogs in comparison to control cells. These findings suggest altered basal autophagy as a pathogenetic mechanism in this novel disease. Lysosomal storage of a specific material or a lysosomal enzyme deficiency were not detected. The genetic groups in the collaborative research where this thesis was part discovered a novel candidate gene for neurodegeneration, ATG4D. In aspects of comparative pathology, altered basal autophagy was uncovered as an additional etiology and morphological differential diagnosis for cytoplasmic vacuolation reminiscent of lysosomal storage diseases in dogs, and functionally linked to the ATG4D variant through this thesis. The importance of basal autophagy in cells was also hereby shown, as disease occurred despite starvation-induced autophagy being functional in the dogs. For veterinary medicine, the characterization of this novel inherited progressive neurodegenerative disease provides a broader scientific knowledge as basis for veterinarians regarding prognostication of affected dogs, and for LR owners regarding breeding. | Avhandlingen beskriver vävnadsförändringarna hos Lagotto Romagnolo (LR) hundar, med en genvariant i ATG4D, som lider av en ny, fortskridande neurologisk sjukdom. Avhandlingens syfte är att undersöka om sjukdomen uppkommer till följd av störd autofagi eller om sjukdomen är en lysosomal inlagringssjukdom. Störd autofagi är en central sjukdomsmekanism i flera neurodegenerativa sjukdomar hos människan, därför är det av intresse att undersöka sjukdomsmekanismer och genetiken bakom liknande sjukdomar hos hundar. Lysosomala inlagringssjukdomar är också ärftliga, beträffar både människor och djur och skadar ofta nervsystemet. Både störd autofagi och störd lysosomal nedbrytning kan leda till att ämnen i stället frigörs som extracellulära vesiklar (EV); små signalbärare som potentiellt kan användas inom diagnostik av störningarna i dessa cellfunktioner. Obduktionsfynden hos 13 beträffade LR hundar bekräftade en enhetlig sjukdomsbild på cellnivå; upplagring av vakuoler i nervcellerna, skador i nervutskotten och nervcellsförlust. Körtelepitelceller och vissa mesenkymala celler uppvisade också cytoplasmiska vakuoler. I immunhistokemiska specialfärgningar syntes ansamling av autofagosomala och lysosomala membranprotein, autofagireceptorer och –last i vävnadsskadorna som tecken på störd autofagi. Med hjälp av elektronmikroskopi och immunelektronmikroskopi kunde delvis samma protein påvisas i de upplagrade vakuolernas membran. Bindvävsceller från beträffade hundar uppvisade förändrad basal autofagi, men oförändrad svält-inducerad autofagi, när autofagi-flödet undersöktes funktionellt i cellkultur. Motsvarade fynd syntes som signifikant flera autofagosomer i bindvävsceller från sjuka LRs under basala förhållanden. Dessutom utsöndrades signifikant flera EV ur de sjuka hundarnas celler under basala förhållanden och förmerat substrat för basal autofagi kunde påvisas i dessa EV. Sammanfattande tyder fynden på störd basal autofagi som en patogenetisk mekanism i sjukdomen. Lysosomal upplagring kunde inte påvisas hos de sjuka hundarna när urin, tre lysosomala enzyms aktivitet och vakuolernas innehåll undersöktes. En ny kandidatgen för nervcellsdegeneration, ATG4D, upptäcktes av genetikerna inom forskningssamarbetet där den här avhandlingen verifierade bristande autofagi hos LR hundar med genvarianten. Avhandlingens bidrag till jämförande patologi är att påvisa störd basal autofagi som en alternativ orsak till vävnadsförändringar som påminner om lysosomal upplagring. En grundläggande relevans för basal autofagi i specifika celler blir också tydlig genom avhandlingen då hundarna har symptom trots att svält-inducerad autofagi fungerar normalt. Inom veterinärmedicin ger avhandlingen, och de kliniska studierna inom forskningssamarbetet, en bredare vetenskaplig basis för veterinärer och LR ägare gällande vårdbeslut, prognosbedömning och rasens avel.

Following the withdrawal of diphenylamine (DPA), 'Irish Bramley' growers lost the facility to store the 'Bramley' for 12 months in controlled atmosphere stores. Manufacturers were grant aided to establish low oxygen stores resulting in a significant advantage over primary producers. Research into improving on farm crop management to reduce losses in storage was undertaken. Farmer perceptions of 'good' and 'poor' orchards under the same management were validated by differences in overall disease levels. Nectria ditissima was found to be the main disease. Acidified water treatments could not cope with the level of soil contamination on bins coming in from the orchard. Geoxe applied as a drench significantly reduced disease levels in storage, more so with wooden bins than plastic.