Structural characterization and functional evaluation of polysaccharides extracted from the heart of date palm (Phoenix dactylifera L.): Insights into α-amylase inhibition and antidiabetic potential.

Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne disease in cattle and other ruminants, is proposed to be at least one of the causes of Crohn disease in humans. MAP and Mycobacterium avium subspecies avium, a closely related opportunistic environmental bacterium, share 95% of their genes and exhibit homologies of more than 99% between these genes. The identification of molecules specific for MAP is essential for understanding its pathogenicity and for development of useful diagnostic tools. The application of gas chromatography, mass spectrometry, and nuclear magnetic resonance led to the structural identification of a major cell wall lipopeptide of MAP, termed Para-LP-01, defined as C20 fatty acyl-D-Phe-N-Me-L-Val-L-Ile-L-Phe-L-Ala methyl ester. Variations of this lipopeptide with different fatty acyl moieties (C16 fatty acyl through C17, C18, C19, C21 to C22) were also identified. Besides the specificity of this lipopeptide for MAP, the presence of an N-Me-L-valine represents the first reported N-methylated amino acid within an immunogenic lipopeptide of mycobacteria. Sera from animals with Johne disease, but not sera from uninfected cattle, reacted with this lipopeptide, indicating potential biological importance.

Structural characterization and rheological behavior of a heteroxylan extracted from Plantago notata Lagasca (Plantaginaceae) seeds

Date palm (Phoenix dactylifera L.) is the major fruit tree grown in arid areas in the Middle East and North Africa. In the last century, dates were introduced to new regions including the USA. Date palms are traditionally propagated through offshoots. Expansion of modern date palm groves led to the development of Tissue Culture propagation methods that generate a large number of homogenous plants, have no seasonal effect on plant source and provide tools to fight the expansion of date pests and diseases. The disadvantage of this procedure is the occurrence of off-type trees which differ from the original cultivar. In the present project we focused on two of the most common date palm off-types: (1) trees with reduced fruit setting, in which most of the flowers turn into three-carpel parthenocarpic fruits. In a severe form, multi-carpel flowers and fruitlets (with up to six or eight carpels instead of the normal three-carpel flowers) are also formed. (2) dwarf trees, having fewer and shorter leaves, very short trunk and are not bearing fruits at their expected age, compared to the normal trees. Similar off-types occur in other crop species propagated by tissue culture, like banana (mainly dwarf plants) or oil palm (with a common 'Mantled' phenotype with reduced fruit setting and occurrence of supernumerary carpels). Some off-types can only be detected several years after planting in the fields. Therefore, efficient methods for prevention of the generation of off-types, as well as methods for their detection and early removal, are required for date palms, as well as for other tissue culture propagated crops. This research is aimed at the understanding of the mechanisms by which off-types are generated, and developing markers for their early identification. Several molecular and genomic approaches were applied. Using Methylation Sensitive AFLP and bisulfite sequencing, we detected changes in DNA methylation patterns occurring in off-types. We isolated and compared the sequence and expression of candidate genes, genes related to vegetative growth and dwarfism and genes related to flower development. While no sequence variation were detected, changes in gene expression, associated with the severity of the "fruit set" phenotype were detected in two genes - PdDEF (Ortholog of rice SPW1, and AP3 B type MADS box gene), and PdDIF (a defensin gene, highly homologous to the oil palm gene EGAD). We applied transcriptomic analyses, using high throughput sequencing, to identify genes differentially expressed in the "palm heart" (the apical meristem and the region of embryonic leaves) of dwarf vs. normal trees. Among the differentially expressed genes we identified genes related to hormonal biosynthesis, perception and regulation, genes related to cell expansion, and genes related to DNA methylation. Using Representation Difference Analyses, we detected changes in the genomes of off-type trees, mainly chloroplast-derived sequences that were incorporated in the nuclear genome and sequences of transposable elements. Sequences previously identified as differing between normal and off-type trees of oil palms or banana, successfully identified variation among date palm off-types, suggesting that these represent highly labile regions of monocot genomes. The data indicate that the date palm genome, similarly to genomes of other monocot crops as oil palm and banana, is quite unstable when cells pass through a cycle of tissue culture and regeneration. Changes in DNA sequences, translocation of DNA fragments and alteration of methylation patterns occur. Consequently, patterns of gene expression are changed, resulting in abnormal phenotypes. The data can be useful for future development of tools for early identification of off-type as well as for better understanding the phenomenon of somaclonal variation during propagation in vitro.

Labdanum resin or "gum" can be obtained from Cistus ladanifer L. by two different extraction methods: the Zamorean and the Andalusian processes. Although its main use is in the fragrance and perfumery sectors, ethnobotanical reports describe its use for medicinal purposes in managing hyperglycemia and mental illnesses. However, data concerning the bioactivities and pharmacological applications are scarce. In this work, it was found that the yield of labdanum resin extracted by the Andalusian process was 25-fold higher than the Zamorean one. Both resins were purified as absolutes, and the Andalusian absolute was purified into diterpenoid and flavonoid fractions. GC-EI-MS analysis confirmed the presence of phenylpropanoids, labdane-type diterpenoids, and methylated flavonoids, which are already described in the literature, but revealed other compounds, and showed that the different extracts presented distinct chemical profile. The potential antidiabetic activity, by inhibition of α-amylase and α-glucosidase, and the potential neuroprotective activity, by inhibition of acetylcholinesterase, were investigated. Diterpenoid fraction produced the higher α-amylase inhibitory effect (~30% and ~40% at 0.5 and 1 mg/mL, respectively). Zamorean absolute showed the highest α-glucosidase inhibitory effect (~14% and ~24%, at 0.5 and 1 mg/mL, respectively). Andalusian absolute showed the highest acetylcholinesterase inhibitory effect (~70% and ~75%, at 0.5 and 1 mg/mL, respectively). Using Caco-2 and HepG2 cell lines, Andalusian absolute and its purified fractions showed moderate cytotoxic/anti-proliferative activity at 24 h exposure (IC50 = 45-70 µg/mL, for Caco-2; IC50 = 60-80 µg/mL, for HepG2), whereas Zamorean absolute did not produce cytotoxicity (IC50 ≥ 200.00 µg/mL). Here we show, for the first time, that labdanum resin obtained by the Andalusian process, and its fractions, are composed of phytochemicals with anti-diabetic, neuroprotective and anti-proliferative potential, which are worth investigating for the pharmaceutical industry. However, toxic side-effects must also be addressed when using these products by ingestion, as done traditionally.

The genus Cedrela is one of the phytochemically rich genera of the family Meliaceae. In this study, two Cedrela species, namely, Cedrela odorata and Toona ciliata M. Roem (formerly Cedrela toona), were selected for in-depth phytochemical profiling with the aid of UPLC-ESI/MSn analysis followed by evaluation of their anti-diabetic potential through assessment of in vitro α-amylase and α-glucosidase inhibitory effects, alongside the molecular docking studies on these target enzymes. UPLC-ESI/MSn technique was applied to tentatively identify the extracts. The anti-diabetic properties were assessed using BioVision α-amylase and α-glucosidase inhibitor screening kits. Further, the molecular docking studies utilized PyRx® and Discovery Studio software. The UPLC-ESI/MSn analysis led to the identification and quantification of 55 metabolites with their fragmentation patterns for the first time for these two species. Flavonoids represented the main identified class, followed by phenylpropanoids, terpenes, tannins, and others. The two species showed potent enzyme inhibition, where C. odorata and C. toona significantly inhibited α-amylase (IC50 = 4.83 ± 0.01 and 3.50 ± 0.03μg/mL) compared to pioglitazone (IC50 = 2.17 ± 0.23μg/mL), while their α-glycosidase inhibitory properties were also potent with (IC50 = 7.17 ± 0.01 and 6.50 ± 0.69μg/mL), respectively, compared to acarbose (IC50 = 4.83 ± 1.02μg/mL). The enzyme inhibitory activities were further confirmed by in silico molecular docking of the main identified components with the respective binding sockets in both α-amylase and α-glycosidase enzymes. These promising results could pave the way for a novel discovery of natural phytoconstituents with potent anti-diabetic activity.

Date palm (Phoenix dactylifera L.) is a globally edible fruit and a traditional dietary component in various cultures. The fruit’s fleshy part is consumed for its nutritional value, while the seeds are discarded or valorized for oil production and as a coffee substitute. The current study aimed to investigate date seeds’ metabolome, in addition to macro- and micro-elements composition within12 major Egyptian cultivars (cvs.) for the first time using gas chromatography coupled with mass spectrometry (GC-MS). Post-silylation GC-MS analysis and headspace coupled with solid-phase microextraction (HS-SPME) were used for nutrients and aroma profiling in roasted seeds, respectively. Furthermore, multivariate data analyses were employed for samples classification and markers identification via principal component analysis (PCA) and orthogonal projection to least square discriminant analysis (OPLS-DA). Models are further validated by permutation test. Moreover, absolute quantification of potential markers was attempted based on reference standards A total of 101 and 65 nutrient and aroma metabolites were annotated, respectively. Fatty acids/esters (38 peaks), sugars (18), organic acids (17), sugar alcohols (7), steroids/triterpenoids (5), alcohols and aldehydes (6), in addition to flavonoids (1) and phenolic acids (3) were identified as major components in GC-MS post-silylation platform. ‘’Khalas’’ cv. seed appeared the most nutritive being enriched in sugars and fatty acids/esters. Moreover, date seed volatiles from different cvs. were dominated by fatty acids/esters (19 peaks), esters (6), and phenols/ethers (9). Anethole (peak 47) was the most abundant at 9.1–23.3% of seeds contributing to their unique aroma, especially ‘’Barhi’’ a premium date cv. PCA score plot of primary metabolites’ dataset revealed for 1-monopalmitin and monostearin as potential markers for ‘’Aref’’ and ‘’Khalas’’. Furthermore, ‘’Barhi’’, ‘’Omeldehn’’, and ‘’Lolo’’ cvs. showed comparable aroma profile and in partial agreement with nutrient results. OPLS-DA model revealed that anethole, estragole, methyl esters of dodecanoic acid and octanoic acid were characteristic in case of ‘’Barhi’’ cv. which are likely to impart a fine aroma and flavor. With regards to minerals, ‘’Zamli’’, ‘’Barhi’’, and ‘’Hasawi’’ cvs. were most rich in calcium, copper, and selenium, respectively. This study offers new perspectives for the phytochemical makeup and valorization potentials of date palm seeds. Fatty acids/esters and sugars were the major components in date palm seeds found enriched in ‘’Khalas’’ cv, while anethole, estragole, methyl esters of dodecanoic acid and octanoic acid were potential markers of ‘’Barhi’’ cultivar. Such extensive profiling identified premium cvs. to be considered for food applications.

Short chain fatty acids (SCFAs) are important regulators of host physiology and metabolism and may contribute to obesity and associated metabolic diseases. Interest in SCFAs has increased in part due to the recognized importance of how production of SCFAs by the microbiota may signal to the host. Therefore, reliable, reproducible, and affordable methods for SCFA profiling are required for accurate identification and quantitation. In the current study, four different methods for SCFA (acetic acid, propionic acid, and butyric acid) extraction and quantitation were compared using two independent platforms including gas chromatography coupled with mass spectrometry (GC-MS) and 1H nuclear magnetic resonance (NMR) spectroscopy. Sensitivity, recovery, repeatability, matrix effect, and validation using mouse fecal samples were determined across all methods. The GC-MS propyl esterification method exhibited superior sensitivity for acetic acid and butyric acid measurement (LOD < 0.01 μg mL-1, LOQ < 0.1 μg mL-1) and recovery accuracy (99.4%-108.3% recovery rate for 100 μg mL-1 SCFA mixed standard spike in and 97.8%-101.8% recovery rate for 250 μg mL-1 SCFAs mixed standard spike in). NMR methods by either quantitation relative to an internal standard or quantitation using a calibration curve yielded better repeatability and minimal matrix effects compared to GC-MS methods. All methods generated good calibration curve linearity (R2 > 0.99) and comparable measurement of fecal SCFA concentration. Lastly, these methods were used to quantitate fecal SCFAs obtained from conventionally raised (CONV-R) and germ free (GF) mice. Results from global metabolomic analysis of feces generated by 1H NMR and bomb calorimetry were used to further validate these approaches.

Protective effect of linseed oil varnish on archaeological wood treated with alum

Chapter 8 - Plant-Derived Antidiabetic Compounds Obtained From African Medicinal Plants: A Short Review

Objective: This work was performed to evaluate the effect of priming with exogenous sources of calcium ion and nitric oxide on the antidiabeticactivity and the alkaloid contents of fenugreek sprouts along with isolation and identification of trigonelline, a bioactive alkaloid responsible for hypoglycemic property of fenugreek.Methods: The fenugreek seeds were pre-treated with calcium chloride (CC), lanthanum chloride (LC) a calcium channel blocker; ethylene glycol-bis (2-aminoethylether) -N, N, N´, N tetra acetic acid (EG) a calcium chelator; sodium nitroprusside (SNP) and 2-(4-carboxyphenyl) -4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CP) a nitric oxide scavenger and germinated for 72 hrs. The sprout extracts were evaluated for their in vitro antidiabetic potential by α-amylase and α-glucosidase inhibition along with their trigonelline content. Trigonelline was isolated from fenugreek sprouts and identified by Infrared analysis and nuclear magnetic resonance (NMR) spectroscopy.Results: The results revealed that sprouts pre-treated with CC and SNP exhibited enhanced antidiabetic potential as well as alkaloid content overcontrol; on the other hand, their action was reversed by their antagonists, EG, LC, and CP. The sprouts pre-treated with 2mM CC showed the best elicitation of alkaloid content and antidiabetic activity followed by SNP-20 mM.Conclusions: The study suggests probable involvement of the signaling molecules, calcium ion, and nitric oxide in pathways associated withbiosynthesis of bioactive compounds responsible for hypoglycemic activity of fenugreek sprouts one of which being trigonelline.Keywords: Antidiabetic, Calcium, Fenugreek sprouts, Nitric oxide, Priming, Trigonelline, Nuclear magnetic resonance.

Phytochemistry, antioxidant and anti-diabetic activities of Sterculia villosa in-vitro

Fatty acid methyl esters from waste beef tallow using supercritical methanol transesterification

The aim of this study was to determine the chemical composition, minerals and antioxidants of heart of date palm from three Saudi cultivars. Moisture (ranged from 80.44% - 82.82%) was the predominant component in the heart date palm. Sukkari cultivar had the highest protein (5.27%) and ash (7.9%) compared to the other two cultivars. Fat was low in the three cultivars. Sucrose was the predominant sugar in the heart date palm and ranged from 7.65% - 20.25%. Potassium was the predominant mineral in the heart date palm and was significantly (p 50 for Solleg, Sukkari and Naboat Saif were 0.12, 0.13 and 0.29 mg/ml, respectively. The results of the present study may highlight the potential importance of heart of date palm as a product rich in essential minerals and antioxidants. Future researches are needed to explore other characteristics of heart of date palm which could be considered for human nutrition.

Microalgae are widely used in the pharmaceutical and energy industries, therefore the conditions for their cultivation and extraction methods play an important role in the profiling and acquisition of lipids. The efficiency of lipid extraction from microalgae has attracted great interest from industry because of the wide variety of lipids and amounts that can be obtained. Acutodesmus obliquus (Scenedesmus obliquus UTEX 393) was used in this study. It was cultivated in Bold 3N medium modified with 75% nitrogen at 25°C, pH 6.8, 125 r/min and a photoperiod of 18/6 h and illuminated with white light provided by a Light-Emitting Diode Surface Mount Device extensions (LED SMD) with an intensity of 1200 µE/(m2•s). The cells were stained with the Red Nile (RN) technique to indicate lipid production. Four extraction methods were compared, classical, microwave (MW), Soxhlet, and ultrasound (US), using the same solvent proportions (hexane: chloroform: methanol=1:2:3). All samples were analyzed with Fourier Transform Infrared Spectroscopy (FTIR) and Gas Chromatography coupled to Mass Spectrometry (GC-MS). The results showed: 1) lipid production detected by RN was consistent with microalgal growth; 2) the MW technique was the best extraction method, according to the statistical analysis through Randomized Complete Block (RCB) design and performance of 4.6%; and 3) the presence of saturated and unsaturated acids was indicated by FTIR spectra. GC-MS was able to identify palmitic and linoleic acids as the likely major constituents of the sample. Keywords: Acutodesmus obliquus, lipid extraction, trans-esterification, Red Nile (RN) DOI: 10.25165/j.ijabe.20181105.3748 Citation: Hurtado D X, Garzon-Castro C L, Cortes-Romero J, Tello E. Comparison of lipid extraction methods for the microalgae Acutodesmus obliquus. Int J Agric & Biol Eng, 2018; 11(5): 211–217.

In this research article, we present a simple and rapid method for synthesis of zinc oxide nanoparticles (ZnONPs) using zinc acetate dehydrate, sodium hydroxide and leaf extract of an important anti-diabetic plant Andrographis serpelifolia (As). The results reveal that UV-visible absorption spectrum indicates peaks at 325 nm which confirms the biosynthesis of ZnONPs. Fourier transform infrared spectroscopy (FT-IR) results indicate the characteristic peak at 511.93 cm−1 for ZnO nanoparticles. EDX analysis revealed the purity of As-ZnONPs and the particle size was between 70 nm and 80 ± 10 nm by both dynamic light scattering analysis (DLS) and Transmission electron microscopy (TEM) analysis. The As-ZnONPs were moderately stable with negative zeta potential value −11.1 mV. Atomic force microscopy (AFM) analysis also revealed the same results as TEM and DLS. The x-ray diffraction (XRD) data revealed that the particles are facets of face-centered cubic crystal structure of ZnO. The As-ZnONPs have exhibited potential antioxidant activity and effective antibacterial activity against gram-positive bacteria. The As-ZnONPs have also potential anti-diabetic activity in vivo (rat model). The antidiabetic activity of As-ZnONPs comparable to that of the standard drug glibenclamide was evidenced by various biochemical parameters like HbA1C, serum lipid profiles, liver and renal functional markers.