Rich Fraction Research Articles

Wild olive fruits: Phenolics profiling, antioxidants, antimicrobial, thrombolytic and haemolytic activities

In this study, wild olive fruits were evaluated for the occurrence of phenolic antioxidant components and valuable nutrients which are distributed wildly in Soon valley of Pakistan. The shade-dried fruits of wild olive were extracted using different solvents to recover phenolic antioxidants. The highest concentration of extractable antioxidant components was recovered from tested fruits using aqueous ethanol compared to other solvents used. Crude concentrated extracts (CCEs) and phenolic rich fractions (PRFs) of tested fruits using hydroxyethanol were found to contain higher amount of total phenolic compounds and total flavonoid compounds along with superior biological attributes. According to ICP-OES analysis, potassium (17.96 g/kg) was the dominant macro element among other identified twenty-five minerals. The tested wild olive fruits juice was found to contain individual natural sugars including galactose (4.92 g/100 g dry weight), sucrose (2.75 g/100 g dry weight), glucose (0.73 g/100 g dry weight); and succinic acid (8.80 mg/100 g of dry matter) as major organic acid when analyzed on HPLC. Oleic acid (47.41 %) was the major monounsaturated fatty acid in the oil extracted from tested fruits. The concentration of phenolic antioxidants and biological activities vary significantly (p < 0.05) among extracting systems used. A strong correlation was also recorded among total phenolic (TP), total flavonoids (TF) and biological attributes of tested wild olive fruits. The results of this study explored wild olive fruits as a propitious source of natural phenolic components and valuable nutrients which reveal its potential use in the development of functional food and nutra-pharmaceuticals.

Polyphenolic rich fraction of Physalis peruviana calyces and its nano emulsion induce apoptosis by caspase 3 up-regulation and G2/M arrest in hepatocellular carcinoma

Phytocompounds found in fruit wastes serve as nature's precious gift to humans in cancer treatment. In this context, the present work aimed to use of Physalis peruviana calyces (PPC) and its nano-emulsion as a novel method in hepatocellular carcinoma treatment. PPC butanolic fraction (PPCBF) was prepared and tested for its total phenol, total flavonoids, and HPLC analysis for phenolics, as well as the in vitro anticancer activity for both normal and nano-emulsion was evaluated. Twenty-four male Sprague-Dawley rats were used; 18 rats were administered intraperitoneally diethylnitrosamine (DEN) once (200 mg/kg body weight) in a saline solution. All nutritional parameters, biochemical parameters, and histopathological examinations were assessed. Results revealed that PPCBF and its nano-emulsion showed high anti-HepG2 activity and induced Pre-G1 apoptosis and cell cycle arrest (G2/M phase). Moreover, PPCBF and its nano-emulsion caused up-regulation of caspase-3 mRNA in HepG2, which could indicate that the intrinsic mitochondrial route triggered apoptosis. All the nutritional and biochemical changes were confirmed by histopathological examination. In conclusion, PPC and its nano-emulsion could be serving as a successful and effective drug from waste with a bright future in the treatment of hepatocellular carcinoma (HCC).

Integrated xylooligosaccharides production from imidazole-treated sugarcane bagasse with application of in house produced enzymes

The application of biorefinery concepts to produce different value-added biomolecules such as xylooligosaccharides (XOs) generates economical competitive, sustainable and environmentally friendly processes. The objective of this work was to develop an efficient imidazole-pretreatment process of sugarcane bagasse (SB) and the use of the obtained hemicellulose fraction in the production of XOs with the application of in house produced xylanolytic enzymes using SB as substrate, under a biorefinery approach. SB imidazole pretreatment allowed the recovery of a hemicellulose rich fraction (34%) with 91.2% of delignification. Xylanase production by Aspergillus niger reached 53.1 U·mL−1 at 120 h. The application of produced xylanases in the enzymatic hydrolysis of extracted xylan, allowed the production of 6.06 g·L−1 of XOs, where xylotriose represented >70%. Great perspectives are viewed for the implementation of mixed processes in a sustainable closed cycle to produce biomolecules with concomitant valorization of subproducts from SB chain.

Melaleuca rugulosa (Link) Craven Tannins: Appraisal of anti-inflammatory, radical scavenging activities, and molecular modeling studies

Ethnopharmacological relevanceGenus Melaleuca or tea tree species are well known to be an important source of biological active oils and extracts. The biological significance appears in their usage for treatment of several clinical disorder owing to their traditional uses as anti-inflammatory, antibacterial, antifungal, and cytotoxic activities.Aim of the study: Our study aimed to investigate the metabolic profile of the M. rugulosa polyphenol-rich fraction along with determination of its anti-inflammatory potential, free radical scavenging and antiaging activities supported with virtual understanding of the mode of action using molecular modeling strategy. Materials and methodsThe anti-inflammatory activity of the phenolic rich fraction was investigated through measuring its inhibitory activity against inflammatory mediators viz tumor necrosing factor receptor-2 (TNF-α) and cyclooxygenases 1/2 (COX-1/2) in a cell free and cell-based assays. Moreover, the radical scavenging activity was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC) and β-carotene assays, while the antiaging activity in anti-elastase, anti-collagenase, and anti-tyrosinase inhibitory assays. Finally, the biological findings were supported with molecular docking study using MOE software. ResultsThe chromatographic purification of the polyphenol-rich fraction of Melaleuca rugulosa (Link) Craven afforded fourteen phytoconstituents (1–14). The anti-inflammatory gauging experiments demonstrated inhibition of inflammatory-linked enzymes COX-1/2 and the TNF-α at low μg/mL levels in the enzyme-based assays. Further investigation of the underlying mechanism was inferred from the quantification of protein levels and gene expression in the lipopolysaccharide (LPS)-activated murine macrophages (RAW264.7) in vitro model. The results revealed the reduction of protein synthesis of COX-1/2 and TNF-α with the down regulation of gene expression. The cell free in vitro radical scavenging assessment of the polyphenol-rich fraction revealed a significant DPPH reduction, peroxyl radicals scavenging, and β-carotene peroxidation inhibition. Besides, the polyphenol-rich fraction showed a considerable inhibition of the skin aging-related enzymes as elastase, collagenase, and tyrosinase. Ultimately, the computational molecular modelling studies uncovered the potential binding poses and relevant molecular interactions of the identified polyphenols with their targeted enzymes. Particularly, terflavin C (8) which showed a favorable binding pose at the elastase binding pocket, while rosmarinic acid (14) demonstrated the best binding pose at the COX-2 catalytic domain. In short, natural polyphenols are potential candidates for the management of free radicals, inflammation, and skin aging related conditions. ConclusionNatural polyphenols are potential candidates for the management of free radicals, inflammation, and skin aging related conditions.

Renoprotective activity of anethole- rich fraction from aromatic herbs on junk food induced diabetic nephropathy in rats

PurposeThis study was carried out to study the effect of anethole rich fraction on the Diabetic Nephropathy (DN) rats, and explore the mechanisms.MethodsMale wistar rats were grouped into 4 (n = 6 per group): Control, junk food fed DN induced rats, low dose and high dose groups. DN was induced by oral junk food feeding. They were monitored for blood glucose levels and urine protein content at regular intervals. At the end of the study, the biological and hematological parameters were measured. Pancreatic and kidney viscera were taken to observe histopathological changes.ResultsBoth the doses of anethole rich fraction could drastically lower blood glucose levels, Low Density Lipoprotein (LDL), Glycated Serum Protein (GSP), Total Cholesterol (TC), Triglycerides (TG) (P < 0.01), Malondialdehyde (MDA) level (P < 0.01 or P < 0.05), increases insulin level (P < 0.01), High Density Lipoproteins (HDL), Glutathione Reductase (GSH) and Superoxide Dismutase (SOD) (P < 0.01 or P < 0.05 or P > 0.05). Both doses of anethole rich fraction also improved the pathological changes of kidney and pancreatic tissues in DN rats (P < 0.01 or P < 0.05 or P > 0.05).ConclusionsHence it can be hypothesized that the high dose of anethole rich fraction (100 mg/kg) could reach the intervention effect and could ameliorate renal damage in DN rats by improving the renal functions, oxidative stress levels, glycometabolism and pathological changes of pancreas and kidney.

Fractionation of carbohydrate polymers from Indonesian sorghum by-products

Extraction of cellulose and hemicellulose from natural resources has attracted considerable interest. In comparison to other cereals, there is relatively little knowledge on the cellulose, hemicellulose and lignin content of sorghum by-products. As such, the aims of this study were to develop a multi-step process for sorghum by-products (bran, stalk and panicles) fractionation and characterise them with regards to their physicochemical properties. In terms of chemical composition, sorghum panicles were found similar to bran, featuring a significant presence of arabinoxylans as well. A sequential alkaline extraction with varying concentrations of NaOH was applied, resulting in the generation of three fractions (residue, hemicellulose and alkali soluble lignin). Fractionation of sorghum stalks with 1.0 M NaOH at 50 °C for 3 h obtained ~64 % (w/w) cellulose rich fraction and 52 % (w/w) of xylose, predominant in the hemicellulose fraction. In the bran, the fractionation with 0.75 M NaOH at 50 °C for 3 h obtained ~40 % (w/w) cellulose rich fraction. Approximately 76 % of the glucose was in hemicellulose fraction reflecting starch solubilisation in the case of bran, while nearly 2 % (w/w) alkaline-soluble lignin was extracted. Overall, this study demonstrated an effective approach for the fractionation and the recovery of key components in sorghum by-products.

Network pharmacology-based study on apigenin present in the methanolic fraction of leaves extract of Cestrum nocturnum L. to uncover mechanism of action on hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the sixth most common type of cancer in the world. It is the third leading cause of cancer-related mortality. In more than 80% of people liver cancer-related death is due to its poor prognosis. The flavonoids obtained from natural sources have potent therapeutic effects on HCC. The flavonoid rich methanolic fraction obtained from ethyl acetate extract of leaf of Cestrum nocturnum (MFLCN) was analyzed by UPLC-QTOFMS/MS for the presence of different flavonoids. The physiochemical and pharmacokinetics properties of the identified flavonoids were performed by absorption, distribution, metabolism, excretion, and toxicity (ADMET). It was selected on the basis of Lipinski rule and hepatotoxicity evaluations. The potential gene dataset of HCC were taken from gene card database and targets compounds were selected from target net prediction. Gene ontology and pathway enrichment analysis of HCC was performed via enricher and David web tools. Cytoscape was used to visualize targets and network pathways. MFLCN contains 33 flavonoids. Among these flavonoids, apigenin was selected as principal active compound on the basis of their pharmacokinetic and ADMET properties. Apigenin has 92 targets out of 627 total targets related to HCC, while there was13 pathways in the target-pathway network. Results revealed that apigenin regulates cell proliferation and survival, primarily through different signaling pathways like estrogen, VEGF, PI3K/AKT1, TNF, FoXO, and Ras signaling pathways. Thus, integrating network pharmacology prediction with m-RNA and human protein atlas validation could be an effective method for understanding the molecular mechanism of apigenin on HCC.

A retrospective study of the relationship between canine age, semen quality, chilled semen transit time and season and whelping rate and litter size

The objectives of this study were to investigate the effect of the dog’s age, semen quality, and the duration and season of semen transit on whelping rate and litter size after insemination with transported chilled extended semen. The sperm rich fraction was collected from 43 dogs of 18 breeds, which were presented at the Clinic for chilled semen transport, in the period from 2017 to 2021. Immediately after collection, the total sperm concentration and count, motility, membrane integrity (HOS test), the percentage of live spermatozoa and sperm morphology (eosin nigrosin staining) were evaluated. The sperm rich fraction was centrifuged and diluted with Tris – fructose - citrate extender with the addition of 20% (v/v) egg yolk, then chilled and prepared for shipping. A dose consisted of at least 200x106 live, motile, morphologically normal spermatozoa. The data on the dog’s age, chilled sperm transit time, the season of transit, and the whelping rate and litter size after insemination were recorded. The whelping rate was 55.8% with a mean (±SEM) litter size of 4.71±0.58 pups. The total number of spermatozoa was higher in artificial insemination (AI) that resulted in whelping compared to unsuccessful AI (P&lt;0.05). No difference was observed for any of other sperm quality parameters tested, such as the dog’s age or season of transit regarding whelping rate or litter size. Transit time significantly affected the whelping rate (P&lt;0.01), at (mean±SEM) 21.50±1.28 and 37.00±5.59 h in successful and unsuccessful AI, respectively. In conclusion, analysis of the factors related to the dogs identified total sperm count and transit time as factors that significantly affected whelping rates in bitches inseminated with transported chilled extended semen.

Breast Milk Lipidome Is Associated With Maternal Diet and Infants' Growth.

ObjectivesThe fatty acid (FA) composition of breast milk is a relevant aspect related to the development of the lactating infant. The present study aimed at exploring correlations between dietary intake of macro- and micronutrients with the FA profile in breast milk, and the possible implication for infants' growth.Study DesignBreast milk samples from a cohort of lactating women were collected 7–15 days postpartum. The FA profiles in triacylglycerol (TAG) and phospholipid (PL)-rich fractions were analyzed by gas chromatography. Diet was registered during the third trimester of pregnancy by means of a food frequency questionnaire (FFQ). In addition, anthropometric measurements of infants were collected from gestation and up to 12 months postpartum.ResultsThe FA profile in breast milk was characterized by a median of 37.4, 41.3 and 16.8% of saturated, monounsaturated, and polyunsaturated FAs, respectively. From the dietary components, zinc, iron, and B group vitamins were correlated positively with the proportion of total n-3 FAs in TAG and C20:5 n-3 in PL. Lycopene, vitamin E, zinc, and vitamin B2 showed a similar correlation with total polyunsaturated fatty acid (PUFA), total n-6 FAs, C20:4 n-6, and C18:2 n-6 in TAG. Regarding food groups, nuts showed the strongest association with several PUFA both in TAG and PL, while the vegetable group was also positively associated with C18:3 n-3. Furthermore, the concentration of linolenic acid (C18:3 n-3) and palmitic acid (C16:0) were positively associated with increased length for age (LFA) and weight for age (WFA) at 12 months compared with birth [ΔLFA −0.16 (−0.85, 0.37); ΔWFA −0.26 (−0.77, 0.21)].ConclusionsMothers' intake of nuts, dietary sources of zinc, iron, and B group vitamins were identified as potential predictors of a high-unsaturated FA profile in breast milk. In addition, linolenic and palmitic acids in breast milk were positively associated with infants' growth in the first year of life.

AlCoCrFeNi-NiTi high entropy alloy composites: Microstructure and wear performance

In the current research, AlCoCrFeNi–x wt% NiTi (x = 0, 5, and 10) high entropy alloy (HEA) composites were prepared using a combination of mechanical alloying and spark plasma sintering methods. Microstructural studies showed that the introduction of 5 wt% NiTi slightly changed the BCC and FCC fractions of the HEA matrix. However, in the HEA-10NiTi sample, which exhibited increase in the interfacial regions and acceleration of interdiffusion between elements, the (Ni,Al)-rich BCC phase fraction was considerably reduced. According to nano-indentation tests, the NiTi and BCC phases possess higher hardness-to-elastic modulus values that confirm the positive effect of these two phases on tribological behavior. Among the prepared specimens, HEA-5NiTi had the lowest wear rate (0.71 ± 0.01 mm3/N·m). This is attributable to the higher BCC phase fraction and reinforcing effect of the NiTi particles.

Exploring the performance of amaranth grain starch and protein microcapsules as β-carotene carrier systems for food applications

Underexploited sources of bio-based wall materials for bioactive compounds (such as β-carotene) encapsulation have gained increasing interest within the scientific community. In this study, the potential of amaranth (Amaranthus cruentus) grain starch and protein rich fractions as microcapsules’ wall materials to carrier β-carotene was evaluated. Microcapsules were produced by spray-drying and their morphological and physicochemical characterisation was carried out. The microcapsules presented a spherical shape (particle size distribution: 0.3–30 µm) and encapsulation efficiencies ranging from 64 % to 69 %. Results showed that protein-based microcapsules had better β-carotene storage stability as compared to starch-based microcapsules (at 8 and 25 °C). β-carotene release kinetics at 37 °C and pH 7.4 could be mainly described by structural-relaxation phenomenon using the linear superposition model. Moreover, encapsulated β-carotene exhibited higher bioaccessibility than its free form after simulated in vitro digestion tests. Microcapsules did not affect cell viability at 0.0625 mg L−1 of β-carotene. Thus, amaranth grain biopolymers-based microcapsules were successfully developed as promising β-carotene delivery systems to be added to food products and consequently, to improve their functionality.

In vitro Evaluation of Polyphenol Rich Fraction of Ipomea batatas (Mother delight) Leaf Extract as Anti-inflammatory Agents and its LCMS Profile

High intake of plant foods is associated with lower risk of chronic diseases as suggested by epidemiological evidence. This study aimed to investigate the anti-inflammatory potential of polyphenol rich fraction of Ipomoea batatas leaf extract and the compounds possibly responsible for the activity using LCMS. The anti-inflammatory studies were carried out in vitro by protein denaturation technique. The result shows significant different (p&lt;0.05) percentage inhibition of denaturation 90.59% at 1000 μg/ml of the extract compared to the standard drug (declofenac) 95.12% at 1000 μg/ml. The LCMS profiling of the extract revealed the presence of many metabolites (polyphenols) such as chrogenic acid, myricetin, furocoumarinic acid, aromadendrin, naringenin, 4,5-dicaffeolquinic acid and abietinol among others. The presence of these metabolites could be the reason for the anti-inflammatory effect of the extract observed. This implies that the plant can be exploited for its medicinal, therapeutic properties and possibly used to reduce the risk of many chronic diseases. Keyword: Anti-inflammatory activity, Protein denaturation technique, Mother delight, LCMS, Phytochemicals.

Effect of Flavonoid Rich Fraction of Coriandrum sativum Leaf on Lipid Profile, Nitric Oxide, Ang II and Cardio Histopathology in L-NAME Intoxicated Experimental Rats

Coriandrum sativum has been used in traditional medicines as an anti-inflammatory, analgesic, and antibacterial agent. It is recently been shown to have antioxidant activities. This study evaluated the effects of Flavonoid Rich Fraction of Coriandrum sativum Leaf (FRFCSL) on Lipid Profile, Nitric Oxide, Ang II and Cardio Histopathology in L-NAME intoxicated Experimental Rats. Standard analytical method and DPPH radical scavenging activity were employed for the phytochemicals and antioxidant analysis respectively. The acute toxicity study showed the extract and flavonoids fraction are nontoxic at 5000mg/kg body weight. Thirty rats were divided into five groups of six (6) rats each. Group 1 was administered water and feed only, Group 2 was administered 40mg/kg N-Nitro-L-Arginine Methyl Ester (L-NAME) without treatment, Group 3 through 5 were administered 40mg/kg N-Nitro-L-Arginine Methyl Ester (L-NAME) with concomitant administration of Captopril (20mg/kg b.w), FRFCSL (200mg/kg b.w) and FRFCSL (400mg/kg b.w) respectively for 21 days. In phytochemical screening, presence of flavonoids, saponins, tannins, alkaloids, cardiac glycosides, steroids were confirmed as part of secondary metabolites in the extract. The flavonoid fraction showed increase % DPPH radical inhibition in concentration-dependent manner. FRFCSL treatment significantly (p&lt;0.05) and dose-dependently lowered low density lipoprotein and triglycerides. Also It significantly (p&lt;0.05) prevented L-NAME induced decrease in serum angiotensin II, high density lipoprotein and serum nitric oxide concentrations compared to the untreated rats. The Flavonoid Rich Fraction of Coriandrum Sativum Leaf showed great potential as antihypertensive, antihyperlipidemic and cardioprotective agent in rats thus confirming its usefulness in traditional health therapy and potential for antihypertensive drug development.

QS42. Closing The Gap on Clinical Translation of Cell-Based Therapies: A Novel Technique For Intraoperative Isolation of the Stromal Vascular Fraction for Bone Tissue Engineering

Purpose: Adipose-derived stems cells (ASC) have demonstrated promise across many areas of regenerative medicine, including bone tissue engineering. One major barrier to clinical translation and broad application of these therapeutics is current protocols which require laboratory cell culture and enzymatic digestion to produce stem cell rich product. More optimal methodologies would circumvent the current protocols, and instead, employ mechanically-based techniques to produce ASC rich stromal vascular fraction (SVF) intraoperatively for immediate clinical application. Therefore, the purpose of this study was to develop a clinically translatable technique for intra-operative harvest, isolation, and implantation of SVF with the primary aim of enhancing bone healing at irradiated fracture sites. Methods: Male Lewis rats (n=29) were divided into groups: Fracture (Fx), Radiation with Fracture (XRT), and Radiation with Fracture and SVF implantation (SVF). Experimental groups received 35Gy of radiation. All groups underwent mandibular osteotomy and external fixation. Inguinal fat pads were minced and serially processed using Tulip Sizing Transfers (2.4mm, 1.4mm, 1.2mm). Serial filtration (800 micron, 400 micron) and centrifugation was performed. The resultant oil and aqueous layers were discarded and the cell pellet was collected for immediate implantation at the osteotomy site. Animals were sacrificed on post-operative day 40. Gross pathology and MicroCT analysis were utilized to determine union rates and the quality of the bone formed at the osteotomy site. Biomechanical strength testing was performed until failure to evaluate yield and ultimate load of new bone at the osteotomy site. Results: Immediate implantation of SVF increased union rates compared to XRT alone (79% vs. 20%). Additionally, MicroCT analysis demonstrated high quality new bone formation in irradiated fractures treated with SVF compared to the control based on bone mineral density (666.2 ± 32.0 vs. 312.2 ± 51.7; p=0.000) and bone volume fraction (0.744 ± 0.072 vs. 0.350 ± 0.041; p=0.000). In fact, implantation of SVF into irradiated fracture sites resulted in bone quality similar to the bone formed at non-irradiated fracture sites, as there was no significant difference found between groups (BMD: 666.2 ± 32.0 vs. 710.3 ± 38.0; p=0.390, BVF: 0.744 ± 0.072 vs. 0.803 ± 0.04; p=0.300). Radiation significantly diminished the biomechanical properties of bone, including yield (23.6 ± 28.2 vs. 81.9 ± 31.3; p=0.002) and ultimate load (33.7 ± 30.9 vs. 87.3 ± 26.7; p=0.005). SVF implantation improved yield (52.0 ± 28.6 vs. 81.9 ± 31.3; p=0.161) and ultimate load (33.7 ± 30.9 vs. 87.3 ± 26.7; p=0.145) of the irradiated bone to the level of the non-irradiated control, as there was no significant difference between groups. Conclusions: Use of the stromal vascular fraction for bone tissue engineering demonstrates great potential, including applications in irradiated fracture healing. In this study, we developed a novel approach that eliminates laboratory dependent techniques and instead, utilizes mechanical methods that would enable intraoperative SVF harvest, isolation, and immediate implantation. While further studies are required to optimize this approach, the results of this study are incredibly promising for the long-awaited translation of cell-based therapeutics into the clinical arena.

Quercetin-Rich Extracts from Onions (Allium cepa) Play Potent Cytotoxicity on Adrenocortical Carcinoma Cell Lines, and Quercetin Induces Important Anticancer Properties.

Adrenocortical carcinoma (ACC) is a rare subtype of cancer, with a poor prognosis in children and adults. Mitotane is the only approved adrenolytic drug for the treatment of ACC, which has controversies regarding its efficacy and side effects on patients. Onion (Allium cepa), a worldwide consumed food, is associated with many health benefits. Along with its glycosides, the flavonoid quercetin is abundant in onions. After evaluating the cytotoxicity of A. cepa extracts on adrenocortical carcinoma cell line (H295R), the rich quercetin fractions had better results. Then, we aimed to compare the quercetin vs. mitotane effectiveness, using adrenocortical carcinoma cell lines H295R and SW-13. Quercetin showed a higher cytotoxicity response on both cancerous cell lines after 10 µM concentration, while mitotane only after 30 µM. Cell cycle dynamics were altered upon quercetin treatments, with G2 phase increase with 30 µM of quercetin on H295R cell line and G1 arrest on SW-13 cell line with 15 µM. Early and late apoptosis, alongside intracellular calcium, were increased on SW-13 treated with 30 µM of quercetin, and ROS rates were reduced by quercetin on H295R. Therefore, quercetin-rich onions have the potential to be a natural source of anticancer agents for adrenocortical carcinoma.

A Metabolomics Approach to Biomarker Discovery Reveals That Tocotrienols Have Potential Beneficial Impact on Maintenance Hemodialysis Population

ObjectivesCardiovascular disease is considered major comorbidity in the maintenance hemodialysis (MHD) population. A dietary supplement such as tocotrienols have shown to improve cardiovascular health in these patients. In our study, we hypothesized that the tocotrienols may possess potential beneficial impacts on MHD patients by improving their metabolomic profile. MethodsChanges in plasma metabolomic profile of the MHD patients with or without tocotrienol rich fraction (TRF) supplements (300 mg/day) for 12 months period on plasma metabolomic profile of the US (n = 85) and Malaysian (n = 80) MHD was investigated using 1H-NMR based metabolomics approach followed by Receiver Operating Characteristics (ROC) curve based biomarker model validation. ResultsPartial linear square discriminant analysis showed a significant difference in the metabolomic profile of the MHD patients between the placebo and the TRF supplemented groups over 12 months in both US and Malaysian cohorts. According to metabolite identification and quantification proline, 4-hydroxyproline, ornithine, glutamate, 4-aminobutyrate, 3-aminoisobutyrate, acetoacetate, and 3-hydroxybutyrate were found to be significantly (p < 0.05) altered between the placebo group and TRF groups over 12 months. The biomarker models created using these key metabolites had shown good predictive accuracy (Area under the ROC curve > 0.8) in both cohorts with high sensitivity (79% and 78%) and specificity (84% and 80%) in the US and Malaysian cohorts, respectively. Further model validation relied on the permutation test with 1000 permutations (p < 0.001), validating the predictive ability of the created biomarker models in both cohorts. The biomarker meta-analysis revealed that trans-4-hydroxyproline, proline, acetoacetate, ornithine, 3-hydroxybutyrate, 3-aminoisobutyrate, and glutamate had similar expression patterns in both cohorts. ConclusionsMetabolites linked to arginine and proline metabolism and synthesis and degradation of ketone bodies pathways were altered in MHD patients over 12 months. TRF modulated these changes in a favorable direction, indicating the potential beneficial impact of TRF supplements in these patients. These metabolites may serve as biomarkers for kidney disease and/or for evaluating the efficacy of TRF supplementation in MHD patients. Funding SourcesPEMANDU, Malaysia

Analysis of Uterine Blood Flow in Breeding Sows through the Estrus and Early Diestrus, and after Artificial Insemination.

This study aimed to determine uterine blood flow indices by transabdominal Doppler ultrasound in sows (n = 18) under different conditions: (i) sows after estrus detection (day 0, D0); (ii) sows 2 h after artificial insemination (AI), performed 24 h after detection of estrus (day 1, D1); (iii) sows in early diestrus (day 5, D5). Moreover, three different types of seminal doses were used for AI depending on the ejaculate fraction included (F1: doses containing only the rich fraction of the ejaculate; F2: F1 + the transition fraction between rich and poor fractions; F3: F2 and poor fraction). The statistical analysis revealed significant differences in some indices regarding the period of analysis (D0, D1, and D5). Diastolic velocity and mean velocity showed lower values at D5 in comparison with D0 and D1 (p < 0.01). On the other hand, the pulsatility index and the relationship systolic velocity/diastolic velocity indicated higher values at D5 in comparison with D0 and D1 (p < 0.01). No differences were observed regarding the type of seminal dose used in any of the time points analyzed (p > 0.05). Neither insemination per se nor the type of ejaculate fraction used immediately modified the uterine vascularity, but some indices are affected by the stage of the estrus cycle (estrus vs. early diestrus).

Lycorine Alkaloid and Crinum americanum L. (Amaryllidaceae) Extracts Display Antifungal Activity on Clinically Relevant Candida Species

Candida species are the main fungal agents causing infectious conditions in hospital patients. The development of new drugs with antifungal potential, increased efficacy, and reduced toxicity is essential to face the challenge of fungal resistance to standard treatments. The aim of this study is to evaluate the in vitro antifungal effects of two crude extracts of Crinum americanum L., a rich alkaloid fraction and lycorine alkaloid, on the Candida species. As such, we used a disk diffusion susceptibility test, determined the minimum inhibitory concentration (MIC), and characterized the components of the extracts using Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (ESI FT-ICR MS). The extracts were found to have antifungal activity against various Candida species. The chemical characterization of the extracts indicated the presence of alkaloids such as lycorine and crinine. The Amaryllidaceae family has a promising antifungal potential. Furthermore, it was found that the alkaloid lycorine directly contributes to the effects that were observed for the extracts and fraction of C. americanum.

Bioactive phenolics fraction of Hedera helix L. (Common Ivy Leaf) standardized extract ameliorates LPS-induced acute lung injury in the mouse model through the inhibition of proinflammatory cytokines and oxidative stress

Hedera helix L. (family Araliaceae) is classified as a conventional plant used as a medicinal product in the cure and prevention of upper respiratory tract inflammation and infection due to its secretolytic and broncholytic effects. Our research was conducted to authenticate the anti-inflammatory effect of ivy leaves extract in the prevention of acute lung injury (ALI) caused by intranasal administration of lipopolysaccharides (LPS). In-vitro antimicrobial, anti-inflammatory, and anti-oxidant were evaluated, in addition to the in-vivo acute lung inflammation model induced by LPS in mice. The animals were divided into seven groups randomly (each group containing 10 mice): control negative (saline only), control positive (LPS group), standard (Dexamethasone 2 mg/kg), ethanolic ivy leaves extract (EIE, 100 mg/kg), ethanolic ivy leaves extract (EIE, 200 mg/kg), saponin rich fraction (SRF, 100 mg/kg) and phenolic rich fraction (PRF, 100 mg/kg). Right lungs were homogenized to determine the levels of SOD, MDA, catalase, IL-10, TNF-α, NO, IL-1β, IL-6, PGE2, and MPO. Left lungs were excised for histopathology and histomorphometry. Immunohistochemistry of Cox-2 and TNF-α levels were measured. Additionally, Western blotting was used to determine the levels of phosphorylated MAPK. Also, the ethanolic extract was also standardized through HPLC analysis for its content of rutin.The data showed that the oral supplementation with EIE, 200 mg/kg significantly (P < 0.05) decreased the pro-inflammatory mediators, and oxidative stress biomarkers induced by LPS. Interestingly, the phenolics showed promising activity, therefore they are responsible for the action. In conclusion, the standardized ivy leaf extract could be advised for acute lung injury for its antimicrobial, anti-oxidant, and anti-inflammatory activities.

Evaluation of immunological adjuvant activities of saponin rich fraction from the fruits of Asparagus adscendens Roxb. with less adverse reactions

The hemolytic activity, in vitro as well as in vivo toxicity, and immunomodulatory potential of saponins-rich fraction of Asparagus adscendens Roxb. fruit (AA-SRF) have been assessed in this study in order to explore AA-SRF as an alternative safer adjuvant to standard Quil-A saponin. The AA-SRF showed lower hemolytic activity (HD50 = 301.01 ± 1.63 µg/ml) than Quil-A (HD50 = 17.15 ± 2.12 µg/ml). The sulforhodamine B assay also revealed that AA-SRF was less toxic to VERO cells (IC50≥200 ± 4.32 µg/ml) than Quil-A (IC50 = 60 ± 2.78 µg/ml). The AA-SRF did not lead to mortality in mice up to 1.6 mg and was much safer than Quil-A for in vivo use. Conversely, mice were subcutaneously immunized with OVA 100 μg alone or along with Alum (200 μg) or Quil-A (10 μg) or AA-SRF (50 μg/100 μg/200 μg) on days 0 and 14. The AA-SRF at 100 μg dose best supported the LPS/Con A primed splenocyte proliferation activity, elevated the serum OVA-specific total IgG antibody, IL-12, CD4 titer and upsurged CD3/CD19 expression in spleen as well as lymph node sections which in turn advocated its adjuvant potential. Thus, AA-SRF can be further studied for use as a safe alternative adjuvant in vaccines.