Biochemical Composition of Pumpkin Seeds and Seed By-Products.

The goal of the current work was to assess the nutritional profile and phytochemical properties of cucurbit (Cucurbita maxima L.) seeds, seed oils and oil extraction by-products (e.g., seed-cakes). Our results suggest a high nutritional value for both cucurbit seeds and cucurbit cake, while γ-tocopherol was the richest compound, with traces of α, ß and δ-tocopherol compounds also detected. Regarding the free sugars composition, there were recorded significant statistical differences between seeds and cucurbit seed-cake, although sucrose content was the highest for both matrices (1.97 and 2.9 g/100 g dw, respectively) followed by trehalose (0.26 and 0.25 g/100 g dw, respectively), fructose (0.20 and 0.34 g/100 g dw, respectively) and glucose (0.21 and 0.19 g/100 g dw, respectively). In terms of organic acids, oxalic was the only compound detected in seed cake (0.006 g/100 g dw), while in seeds only traces of oxalic and malic acid were detected. In relation to fatty acid composition, linolenic acid was the most abundant compound in both seeds and seed-cake (43.9% and 41.5%, respectively), while oleic acid (37.0% and 36.3%, respectively), palmitic acid (12.2% and 14.0%, respectively) and stearic acid (4.83% and 5.46%, respectively) were detected in lesser amounts. Moreover, polyunsaturated fatty acids (PUFA) were the major fatty acids class (44.5% and 42.3% in seeds and seed cake, respectively) compared to monounsaturated fatty acids (MUFA; 37.4% and 36.7% in seeds and seed cake, respectively) and saturated fatty acids (SFA; 18.1% and 21.0% in seeds and seed cake, respectively) which were detected in lower amounts. Furthermore, the tested extracts did not present any cytotoxic or hepatoxic activity at the maximum tested concentration (GI50 > 400 µg/mL), while seed oils presented satisfactory antimicrobial properties with inhibitory activity against the studied bacterial strains and fungi. Our findings provide valuable knowledge regarding the exploitation of pumpkin seeds and seed by-products as valuable natural sources of nutrients and phytochemicals in the food industry sector within the context of a circular economy.

Seed Priming and Biopriming in Two Squash Landraces (Cucurbita maxima Duchesne) from Tunisia: A Sustainable Strategy to Promote Germination and Alleviate Salt Stress.

In recent years, seed priming has gained interest, with researchers aiming to enhance seed germination and early growth, especially under abiotic stress conditions. In this study, seeds from two squash landraces (Cucurbita maxima Duchesne; i.e., Galaoui large seeds (Galaoui hereafter) and Batati green (Batati hereafter)) were subjected to different priming methods ((a) 0.3% and 0.4% KNO3 (halopriming); (b) 0.1% and 0.2% GA3 (hormopriming); (c) inoculation with Trichoderma spp. (T. harzianum, T. viride, and T. virens), Bacillus subtilis, and Pseudomonas fluorescens (biopriming) in order to promote germination parameters and seedling growth under salinity stress (0, 100, and 200 mM of NaCl). Our findings indicate the better performance of primed seeds compared to the untreated ones in terms of germination and seedling growth traits, although a varied response depending on the priming method and the landrace was observed. The highest germination percentage (GP) and the lowest mean germination time (MGT) were observed in 0.4% KNO3-primed seeds. The positive effects of 0.4% KNO3 were also depicted in all traits related to seedling growth and the seedling vigor index (SVI), indicating its effectiveness as a priming agent in squash seeds. Under salinity stress conditions, priming with 0.4% KNO3 significantly improved the germination and seedling growth traits for both landraces, while the application of 0.2% GA3 at high salinity significantly improved photosynthetic quantum yield (Fv/Fm ratio). Regarding the effects of biopriming in germination and seedling growth traits, our results indicate that T. harzianum and B. subtilis were the most effective bioagents in promoting germination and seedling growth in Galaoui and Batati seeds, respectively. In conclusion, our findings provide important information regarding the practice of using priming and biopriming agents to enhance the germination and seedling growth capacity of squash seeds, as well to mitigate the negative effects of salinity stress at the critical stages of germination and early growth.

Comparing the differences in quality profiles and antioxidant activity in seven pumpkin cultivars (Cucurbita moschata and Cucurbita maxima) at harvest and during postharvest storage.

Pumpkin, nutritious vegetable, is renowned for its extended shelf life. In this study, seven pumpkin cultivars from Cucurbita moschata and Cucurbita maxima were comparatively characterized for 25 physiochemical quality factors, starch granule structures, antioxidant activity, and correlations at 0-60 days of postharvest (dop). The findings revealed that sucrose and carotenoid contents increased in C. moschata, while they initially increased and then decreased in C. maxima. Additionally, acidity, primarily driven by malic acid, decreased in C. maxima but increased in C. maxima. The starch content of C. moschata and C. maxima reached its maximum value at 30 dop and 20 dop, respectively. The DPPH radical scavenging activity correlated with the carotenoid content in both pumpkin species. Conclusively, C. moschata demonstrated improved nutritional and quality at 20-30 dop, while C. maxima exhibited higher commercial suitability at 10-20 dop. The findings suggested that pumpkin storage was crucial for quality improvement.

Diversity and biological traits of bees visiting flowers of Cucurbita maxima var. zapallito differ between biodiversity-based and conventional management practices.

Low diversity of pollinators and the modified composition of functional groups of bees have been proposed as the causes of pollination deficiency in cultivated Cucurbitaceae species. Functional groups of bees are determined by traits, such as body size, nesting site, and social behavior. The presence of bees with specific traits can be differentially affected by agricultural management practices. This work aimed to assess how management types (agroecological and conventional) in Cucurbita maxima var. zapallito crops affect the abundance of bees with different biological traits. The study was conducted on four farms located in horticultural areas of central-eastern Santa Fe province, Argentina. A total of 108 10-min censuses were conducted to record bee species abundance in flowers. The species were assigned to categories for each of the three biological traits. A total of 552 individuals, belonging to 16 bee species, were recorded. Honey bees were more abundant under conventional management, whereas the native bees Eucera fervens and other species were more abundant under agroecological management. Species of the categories analyzed (body size: small, medium, and large; nesting site: above-ground cavities or ground-nesting; and social behavior: solitary or social) were present on farms under both management types. We found that management type affected bees, and their effects differed among bees with specific biological traits. Medium-sized and small bees, ground-nesting bees, and solitary bees were found in greater abundance on agroecological farms than on conventional farms. Our data allowed us to explain the diversity and abundance of bees relative to the management type and biological traits of the species. Implications for insect conservation: This study suggests that incorporating biodiversity-based management strategies might increase abundance and richness of native bees with different biological traits, ensuring the free pollination service they provide and a taxonomically and functionally diverse assemblage.

Genome-Wide Identification and Characterization of the WRKY Gene Family in Cucurbita maxima.

In higher plants, WRKY transcription factors are broadly involved in a variety of life activities and play an important role in both biotic and abiotic stress responses. However, little is known about the functions of WRKY genes in the popular species, such as Cucurbita maxima (pumpkin), which is planted worldwide. In the present study, 102 CmWRKY genes were identified in the C. maxima genome. Chromosome location, multiple sequence alignment, phylogenetic analysis, and synteny analysis of the CmWRKYs were performed. Notably, we found that silencing CmWRKY22 promoted cucumber mosaic virus (CMV) infection, whereas overexpression of CmWRKY22 inhibited the CMV infection. Subsequently, an electrophoretic mobility shift assay (EMSA) confirmed that CmWRKY22 was able to bind to the W-box at the promoter of CmPR1b, which is a responsive gene of the salicylic acid (SA) signaling pathway. In summary, this study has provided a foundation for the antiviral functions of WRKY transcription factors in C. maxima.

Effects of including of Japanese Pumpkin Seeds and Pomace in the Diets of Pacific White Shrimp (Penaeus vannamei).

A 60-day feeding trial was conducted to evaluate the effects of including pumpkin seeds and pomace in the diets of Pacific white shrimp Penaeus vannamei, and the effects of these supplements on growth performance, body composition, and total polyphenol, flavonoid and carotenoid contents, as well as on total antioxidant activity, and body color parameters. Five diets were evaluated: pumpkin seeds (PS) at 50 and 100 g·kg-1, pumpkin pomace (PP) at 50 and 100 g·kg-1, and a control treatment. Pacific white shrimp (P. vannamei) juveniles (0.60 ± 0.01 g) were stocked in 15 tanks (310 L), containing 30 shrimps per tank, and the treatments were randomly distributed in triplicate. At the end of the experiment, shrimps were euthanized, weighed, and dissected for further analyses. The inclusion of PS in the diets impaired growth performance, reduced the total flavonoid content and had a pro-oxidative effect on muscle. The inclusion of PP in the diets did not affect growth performance, improved the feed conversion ratio, increased the total flavonoid content in the diets and hepatopancreas, and improved the antioxidant activity of the feeds and shrimp muscle. The total carotenoid content of the feeds increased with the inclusion of PS or PP in the diets; however, the total carotenoid content of shrimp increased only in those fed PP diets. Shrimp fed with PS diets showed a yellowish color and higher saturation when fresh and a reddish color and yellow hue angle after cooking. Shrimp fed PP diets turned reddish and yellowish, both when fresh and after cooking. The inclusion of PS in P. vannamei diets is not recommended; however, PP can be included at 100 g·kg-1 without affecting the growth parameters. Further studies evaluating the inclusion of higher PP levels in shrimp diets are recommended.

Effects of drought stress induced by D-Mannitol on the germination and early seedling growth traits, physiological parameters and phytochemicals content of Tunisian squash (Cucurbita maximaDuch.) landraces.

Introduction: Drought stress is one of the most devastating environmental stressors, especially in the arid and semi-arid regions of the world. Considering the major constraints that drought stress poses to crop production and the consequent yield losses in food crops, breeding for climate-resilient crops is an efficient means to mitigate stress conditions. Materials and methods: This study aimed at evaluating the response of four squash (Cucurbita maxima Duchesne) landraces to drought stress at germination and at plant stage. Drought stress was induced by different concentrations of D-mannitol (-0.24, -0.47 and -0.73 MPa). The tested parameters at germination stage included germination percentage, seedling vigor index, seed water absorbance and seedling growth potential. At the plant stage, leaf chlorophyll and carotenoids content, chlorophyll fluorescence, evapotranspiration, photosynthesis activity and several biomarkers, namely malondialdehyde, proline, total phenols content, total flavonoids content and DPPH radical scavenging activity were evaluated in both roots and leaves. Results and discussion: Our results indicate a magnitude of drought stress effects reflected via repression of germination and seedling growth as well as adjustments in physiological functions at later growth stages, in a genotype depended manner. Among landraces, "751" and "746" showed better performance, as evidenced by higher seed germination and seedling growth potential even at high stress levels (-0.47 and - 0.73 MPa), whereas "747" was the most sensitive landrace to drought stress at both tested stages. In conclusion, our findings highlight the importance of squash landraces selection for the identification of elite genotypes with increased tolerance to drought stress.

Metabolomics and Transcription Profiling of Pumpkin Fruit Reveals Enhanced Bioactive Flavonoids and Coumarins in Giant Pumpkin (Cucurbita maxima).

Atlantic giant (AG, Cucurbita maxima) is a type of giant pumpkin in the Cucurbitaceae family and has the world's largest fruit. AG possesses excellent ornamental and economic value due to its well-known large fruit. However, giant pumpkins are usually thrown away after viewing, thus generating a waste of resources. To explore the additional value of giant pumpkins, a metabolome assay was performed between AG and Hubbard (a small fruit pumpkin) fruits. We found that bioactive compounds, especially flavonoids (including 8-prenylnaringenin, tetrahydrocurcumin, galangin, and acacetin) and coumarins (including coumarin, umbelliferone, 4-coumaryl alcohol, and coumaryl acetate), with extensive antioxidant and pharmacological functions, showed higher accumulation in AG fruit than in Hubbard fruits. Comparative transcriptomics of the two pumpkin fruits indicated that the differentially expressed genes (DEGs) encoding PAL, C4H, 4CL, CSE, HCT, CAD, and CCoAOMT were relatively highly expressed, which promoted an increased accumulation of the identified flavonoids and coumarins in giant pumpkins. In addition, the construction of a co-expression network and cis-element analysis of the promoter demonstrated that differentially expressed MYB, bHLH, AP2, and WRKY transcription factors might play vital roles in regulating the expression of DEGs involved in the biosynthesis of several flavonoids and coumarins. Our current results provide new insights into the accumulation of active compounds in giant pumpkins.

Nutritional characterization and antioxidant properties of various edible portions of Cucurbita maxima: A potential source of nutraceuticals.

Pumpkin (Cucurbita maxima) is a widely grown vegetable in Bangladesh and is known as the sole supplier of various nutrients. Many studies evidence the nutritional value of flesh and seed while peel, flower, and leaves were reported scarcely with limited information. Therefore, the study aimed to investigate the nutritional composition and antioxidant properties of flesh, peel, seed, leaves, and flowers of Cucurbita maxima. The seed had a remarkable composition of nutrients and amino acids. Flowers and leaves possessed higher content of minerals, phenols, flavonoids, carotenes, and total antioxidant activity. The order of IC50 value (peel > seed > leaves > flesh > flower) indicates higher DPPH radicals scavenging activity of the flower. Moreover, a significant positive relationship was observed among these phytochemical constituents (TPC, TFC, TCC, TAA) and DPPH radicals scavenging activity. It could be concluded that these five parts of the pumpkin plant have an intense potency to be an exigent component of functional food or medicinal herbs.

Interactions between integrated pest management, pollinator supplementation, and normalized difference vegetation index in pumpkin, Cucurbita maxima (Cucurbitales: Cucurbitaceae), production.

Sustainable production of pumpkin (Cucurbita maxima Duchesne) partly relies on integrated pest management (IPM) and pollination services. A farmer-managed field study was carried out in Yatta and Masinga Sub-Counties of Machakos County, Kenya, to determine the effectiveness of a recommended IPM package and its interaction with stingless bee colonies (Hypotrigona sp.) for pollinator supplementation (PS). The IPM package comprised Lynfield traps with cuelure laced with the organophosphate malathion, sprays of Metarhizium anisopliae (Mechnikoff) Sorokin isolate ICIPE 69, the most widely used fungal biopesticide in sub-Saharan Africa, and protein baits incorporating spinosad. Four treatments-IPM, PS, integrated pest and pollinator management (which combined IPM and PS), and control-were replicated 4 times. The experiment was conducted in 600 m2 farms in 2 normalized difference vegetation index (NDVI) classes during 2 growing seasons (October 2019-March 2020 and March-July 2020). Fruits showing signs of infestation were incubated for emergence, fruit fly trap catches were counted weekly, and physiologically mature fruits were harvested. There was no effect of IPM, PS, and NDVI on yield across seasons. This study revealed no synergistic effect between IPM and PS in suppressing Tephritid fruit fly population densities and damage. Hypotrigona sp. is not an efficient pollinator of pumpkin. Therefore, we recommend testing other African stingless bees in pumpkin production systems for better pollination services and improved yields.

Valorization of Pumpkin Peel as a Source of Bioactive Compounds: Optimization of Heat- and Ultrasound-Assisted Extraction.

The peels from three pumpkin genotypes cultivated in Greece were assessed for their phenolic content and bioactive properties to obtain extracts with a high preservative capacity. The optimization of the extraction was performed through response surface methodology (RSM) based on a Box-Behnken experimental design after applying two extraction techniques: heat-assisted (HAE) and ultrasound-assisted (UAE) extraction. The implemented independent variables were time, solvent concentration, and temperature/power (for HAE/UAE), while as dependent variables the dry residue (DR), reducing power (RP), and total phenolic content (TP) were considered. In general, HAE was the most effective technique for 'TL' (75 min; 30 °C; 24% ethanol) and 'Voutirato' (15 min; 30 °C; 10% ethanol), while UAE was more effective for 'Leuka Melitis' (5 min; 400 W; 0% ethanol). The extracts obtained in the global optimum conditions for each genotype peel were then assessed for their phenolic profile, by HPLC-DAD-ESI/MS, and bioactive potential. Seven phenolic compounds were detected, including four flavonoids, two phenolic acids, and one flavan-3-ol. The extracts presented high antioxidant, antibacterial, and antifungal potential, with no cytotoxicity for non-tumor cells. The optimized conditions for the extraction of preservative compounds from bioresidues were defined, allowing the acquisition of antioxidant and antimicrobial extracts and proving their potential for food application.

Genome-Wide Identification of WD40 Proteins in Cucurbita maxima Reveals Its Potential Functions in Fruit Development.

WD40 proteins, a super gene family in eukaryotes, are involved in multiple biological processes. Members of this family have been identified in several plants and shown to play key roles in various development processes, including acting as scaffolding molecules with other proteins. However, WD40 proteins have not yet been systematically analyzed and identified in Cucurbita maxima. In this study, 231 WD40 proteins (CmWD40s) were identified in C. maxima and classified into five clusters. Eleven subfamilies were identified based on different conserved motifs and gene structures. The CmWD40 genes were distributed in 20 chromosomes; 5 and 33 pairs of CmWD40s were distinguished as tandem and segmental duplications, respectively. Overall, 58 pairs of orthologous WD40 genes in C. maxima and Arabidopsis thaliana, and 56 pairs of orthologous WD40 genes in C. maxima and Cucumis sativus were matched. Numerous CmWD40s had diverse expression patterns in fruits, leaf, stem, and root. Several genes were involved in responses to NaCl. The expression pattern of CmWD40s suggested their key role in fruit development and abiotic stress response. Finally, we identified 14 genes which might be involved in fruit development. Our results provide valuable basis for further functional verification of CmWD40s in C. maxima.

Biodegradable scaffolds based on plant stems for application in regenerative medicine.

Several synthetic and natural materials have been studied for the confection of temporary grafts for application in regenerative medicine, however, the development of a material with adequate properties remains a challenge, mainly because its degradation kinetics in biological systems. Nature provides materials with noble properties that can be used as such for many applications, thus, taking advantage of the available morphology and assembled structures of plants, we propose to study the vegetable stems for use as temporary graft. Since thein vivodegradation is maybe one of the most important features of the temporary grafts, here we have implanted the plant stems from pumpkin, papaya, and castor into the subepithelial tissue of animals and followed their biodegradation process and the local inflammatory response. Mechanical tests, FTIR and contact angle with water were also analysed. The results indicated the mechanical properties and the contact angle were adequate for use in regenerative medicine. The results of thein vivostudies indicated a beneficial inflammatory process and a gradual disintegration of the materials within 60 days, suggesting the plants stems as new and potential materials for development of grafts for use in the field of regenerative medicine.

Effect of pretreatments and solar tunnel dryer zones on functional properties, proximate composition, and bioactive components of pumpkin (Cucurbita maxima) pulp powder.

Pumpkin fruits are large in size and need to be cut into pieces for use. The quality and storage life of cuts rapidly deteriorated. Big size fruits can be converted to shelf stable product to minimize postharvest loss using solar driers. However, drying temperature and RH in long solar tunnel drier not uniform and may affects quality of dried products. Therefore this research work aimed to investigate the effects of pre-drying treatments and solar tunnel dryer zones on the functional properties, proximate composition, and bioactive components of pumpkin pulp powder. Three groups of pumpkin slices were pre-treated in 1% citric acid (20 min), 2% salt (20 min) solutions, and the other group blanched at 65 °C in 1% salt solution (2 min), untreated sample used as a control. Pre-treated samples then dried in three zones of tunnel solar drier (zone I, zone II and zone III). Treatment combinations were laid down in factorial RCBD replicated 3 times. Results showed that pulp powder from salt pre-treated slice and dried at zone III results in the highest values of shrinkage, rehydration ratio, water holding capacity, and bulk density. Moisture content decreased from zone I to III, and with salt blanching in range of 8.2 to 6.4%, no effect in crude fat content, slices pre-treated in 2% salt solution results is better crude protein and fiber contents in zones. Better retention of total polyphones, beta-carotene, ascorbic acid, with high DPPH scavenging activity and lowest IC50 values were observed for salt pre-treated sample but dried in zone II of the drier. In general relatively better functional properties, proximate composition and bioactive compounds of the powder can be preserved when slices pre-treated in 2% salt solution and dried in zone II of solar tunnel drier characterized by 54.9 ± 3.7 °C, RH value of 31.4 ± 3.4% and air velocity of 0.45 m/s.

Whole-genome resequencing identified QTLs, candidate genes and Kompetitive Allele-Specific PCR markers associated with the large fruit of Atlantic Giant (Cucurbita maxima).

Atlantic Giant (AG) pumpkin (Cucurbita maxima) produces the world's largest fruit. Elucidating the molecular mechanism of AG fruit formation is of scientific and practical importance. In this research, genome-wide resequencing of an F2 population produced by a cross between AG and its small-fruit ancestor Hubbard was used to identify quantitative trait loci (QTLs) and candidate genes. Transgressive segregation of fruit size-related traits was observed in the F2 population, suggesting that fruit size was a quantitative trait controlled by multiple genes. A genetic map with an average physical distance of 154 kb per marker was constructed, and 13 QTLs related to fruit size were identified using bin-map construction. RNA sequencing analysis revealed that pathways associated with assimilate accumulation into the fruit, including carbohydrate metabolism, were significantly enriched in differentially expressed genes. According to the predicted impact of mutation on the biological function of certain proteins, 13 genes were selected as candidate genes associated with fruit size, among which two phytohormone-related genes, CmaCh17G011340 (a flavin-containing monooxygenase) and CmaCh04G029660 (a leucine-rich repeat protein kinase) were chosen for further investigation. Finally, one insertion-deletion (inDel) and three single nucleotide polymorphisms (SNPs) were successfully transformed to Kompetitive Allele-Specific PCR (KASP) markers. The novel QTLs and candidate genes identified provide insights into the genetic mechanism of large fruit formation of AG, and the genetic map and tightly linked KASP markers developed in this study can be employed for marker-assisted breeding to alter fruit size of C. maxima.

Genome-Wide Identification and Characterization of Members of the ACS Gene Family in Cucurbita maxima and Their Transcriptional Responses to the Specific Treatments.

Ethylene biosynthesis and signal transduction play critical roles in plant sex differentiation. ACS (1-aminocyclopropane-1-carboxylic acid synthase) is a rate-limiting enzyme in ethylene biosynthesis. However, the understanding of the ACS gene family in Cucurbita maxima is limited. Here, we identified and characterized 13 ACS genes in the C. maxima genome. All ACS genes could be divided into three groups according to a conserved serine residue at the C-terminus. Thirteen CmaACS genes were found to be randomly distributed on 10 of the 20 chromosomes of C. maxima. The ACS gene exhibits different tissue-specific expression patterns in pumpkin, and four ACS genes (CmaACS1, CmaACS4, CmaACS7, and CmaACS9) were expressed specifically in both the female and male flowers of C. maxima. In addition, the expression levels of CmaACS4 and CmaACS7 were upregulated after ethephon and IAA treatments, which ultimately increased the number of female flowers, decreased the position of the first female flower and decreased the number of bisexual flowers per plant. These results provide relevant information for determining the function of the ACS genes in C. maxima, especially for regulating the function of ethylene in sex determination.

In Silico Study of Cucurbita maxima Compounds as Potential Therapeutics Against Schistosomiasis.

Schistosomiasis, a disease usually related to poverty and poor sanitation, affects more than 200 million people worldwide. Since the 1970s, the medical sector has depended on a single drug, praziquantel, for the treatment of the disease. The emerging evidence of resistance of the Schistosoma parasite to praziquantel and the drug's inefficacy against juvenile stages of the parasite makes the need to find alternative drugs an urgent matter. In this study, we explored the inhibition potential of compounds from Cucurbita maxima using molecular docking studies on Schistosoma mansoni purine nucleoside phosphorylase (SmPNP) and Schistosoma haematobium 28-kDa glutathione S-transferase (Sh28kDaGST). Following molecular docking studies and analysis of the active sites, the primary amino acids that were observed and shown to be involved in the SmPNP-ligand interaction are CYS 33, ARG 86, HIS 88, TYR 90, ALA 118, ALA 119, PRO 200, TYR 202, GLU 203, VAL 219, MET 221, THR 244, ASN 245, PRO 257 and HIS 259. For the Sh28dKa-ligand interaction, the primary amino acids were PHE 11, ARG 16, TRP 41, LEU 53, GLU 70 and SER 71. Momordicoside I aglycone binds to SmPNP with the lowest binding affinity of -7.9 kcal/mol by pi sigma bond interactions with HIS 88. Balsaminoside B binds to Sh28kDaGST with a binding affinity of -7.6 kcal/mol by hydrogen bond interaction with TRP 41, LEU 53 and SER 71. Pharmacokinetic studies showed favourable drug-like properties for the 10 compounds that exhibited the lowest binding energies. Therefore, we propose that bioactive compounds from C. maxima be considered as potential novel drug hits in the treatment of schistosomiasis.

Development of an Extraction Method of Aflatoxins and Ochratoxin A from Oral, Gastric and Intestinal Phases of Digested Bread by In Vitro Model.

Validated extraction methods from in vitro digestion phases are necessary to obtain a suitable bioaccessibility study of mycotoxins in bakery products. The bakery industry produces bread with different ingredients to enrich the nutritional properties of this product and protect it from fungal growth. This bread can be contaminated by AFB1, AFB2, AFG1, AFG2 and OTA, so an extraction method was developed to analyse these five legislated mycotoxins in digested phases of two types of bread, one with wheat and the other with wheat and also enriched with Cucurbita Maxima Pepo at 20%. The studied "in vitro" digestion model consists of oral, gastric and duodenal phases, each one with different salt solutions and enzymes, that can affect the extraction and most probably the stability of the mycotoxins. The proposed method is a liquid-liquid extraction using ethyl acetate by extract concentration. These analytes and components have an important effect on the matrix effect (MEs) in the analytical equipment, therefore, validating the method and obtaining high sensitivity will be suitable. In the proposed method, the highest MEs were observed in the oral phase of digested pumpkin bread (29 to 15.9 %). Regarding the accuracy, the recoveries were above 83% in the digested duodenal wheat bread and above 76 % in the digested duodenal pumpkin wheat bread. The developed method is a rapid, easy and optimal option to apply to oral, gastric and duodenal phases of digested bread contaminated at a level of established maximum levels by European legislation (RC. 1881/2006) for food.

Potential application of light element stable isotope ratio in crude fiber for geographical origin verification of raw and cooked kabocha pumpkin (Cucurbita maxima).

In this study, we investigated the possibility of the geographical origin discrimination between Japanese and Mexican kabocha pumpkin using δ13C and δ18O in lyophilized raw flesh. Kabocha harvested in Hokkaido, the major kabocha production area in Japan, could be discriminated against not only that grown in Mexico but also that in New Zealand. However, seasoning after cooking or processing affected the δ13C and the δ18O values. Crude fiber extraction eliminated the effect of seasonings and enabled the adoption of the δ13C and δ18O values in crude fiber for the geographical origin verification of kabocha even after cooking and processing. The usage of δ13C and the δ18O in crude fiber would extend the application possibilities of stable isotope analysis in the geographical origin determination of various cooked and processed vegetables and fruits as well as raw materials.

The computationally predicted drug-likeness, pharmacokinetics properties, medicinal chemistry parameters, and toxicity properties of Cucurbita maxima compounds.

Natural compounds are increasingly becoming an important source of drug leads for computer-aided drug design approaches. Cucurbita maxima has been observed to have medicinal properties and can, therefore, be a potential source of novel drug leads. However, before compounds can be synthesized in the lab for tests, modern approaches require that the candidate compounds be screened for drug-likeness characteristics and toxicity, among others. In this work, the computational tools, SwissADME and DataWarrior were used to screen C. maxima compounds for their potential consideration as drug leads. A total of 130 compounds, downloaded from the LOTUS natural products database, were computationally analysed. The data set presented in this work will be useful to researchers searching for novel drug leads based on natural compounds.