Fraction F1 Research Articles

Evaluation of herbicidal potential of Siderophores produced by Amycolatopsis lurida strain 407

The urgent need for sustainable agriculture has intensified the search for environmentally friendly alternatives to chemical herbicides. This study investigates the herbicidal potential of siderophores produced by Amycolatopsis lurida strain 407, focusing on its effects on the growth of ryegrass and redroot weeds. Strain 407 exhibited two distinct colony morphologies—red and white—when cultured under varying environmental conditions. The cell-free culture filtrate (CFCF) from both colony types significantly inhibited the growth of ryegrass and redroot. The concentration of siderophore produced in the iron-deficient medium was measured to be 613.4 ppm for 407 red and 388.5 ppm for 407 white, which indicates significant iron chelating activity. This study also showed a direct relationship between the presence of siderophore in plant culture medium and reduced growth. Also, analysis of fractions of the aqueous phase resulting from column chromatography revealed that all fractions from the 407 red reduced ryegrass shoot length by up to 45% and root length by 83–86%, while redroot seedling length decreased by up to 36%. Fractions from 407 white completely inhibited germination or reduced ryegrass root length by up to 94% and redroot seedling length by 52%. Fractions F4 W to F7 W and F2 R to F8 R, which showed iron chelating activity were most effective in reducing plant growth, suggesting that there are metabolites, alone or in company with siderophores, synergistically do herbicidal activity. The innovative application of siderophores as bioherbicide presents a promising environmentally friendly alternative to chemical herbicides.

Exploring the traits and possible ecological risks of heavy metals in biochars derived from rice husk and sugar beet pulp.

This research investigated the properties and potential environmental hazards associated with biochars derived from rice husk (RH) and sugar beet pulp (SBP), both of which are rich in heavy metals (HMs). The results indicated that the concentration of various HM fractions is significantly affected by the type of feedstock and the pyrolysis temperature. Specifically, the total concentrations of HMs in biochars produced at 600°C were found to be 10-140% higher than those in the original biomasses, a phenomenon attributed to the precipitation of HMs. Cd was a notable exception, exhibiting a reduction of 3-7% in the resultant biochars when compared to biomass, likely attributable to its volatilization. The results also revealed that the F1 + F2 fraction of HMs were largely transformed into F3 + F4 fraction during combustion, indicating that pyrolysis may reduce the ecotoxicity of HMs present in contaminated biomass. However, the process did not effectively diminish the F1 and F2 fractions of Cr and Cd. Elevated pyrolysis temperature significantly enhanced the reduction of HMs phytoavailability. Specifically, the phytoavailability of HMs in biochars produced at 600°C exhibited a decrease ranging from 10 to 92% when compared to the original biomass. Conversely, an unexpected rise in the phytoavailable fractions of Cr and Cd was noted in both RH and SBP biochars as the pyrolytic temperature increased, which correspondingly raised the potential ecological risk index (PERI). All materials analyzed exhibited a very high risk level, with PERI values exceeding 800, primarily due to the significant toxicity of Cd. Excluding Cd from consideration, the biomasses and their resultant biochars displayed PERI values ranging from 7 to 13. It is important to acknowledge that pyrolysis may not effectively diminish the environmental toxicity associated with HMs present in contaminated biomass, thereby limiting its safe application.

Aqueous Extracts of Rhus trilobata Inhibit the Lipopolysaccharide-Induced Inflammatory Response In Vitro and In Vivo.

Chronic noncommunicable diseases (NCDs) are responsible for approximately 74% of deaths globally. Medicinal plants have traditionally been used to treat NCDs, including diabetes, cancer, and rheumatic diseases, and are a source of anti-inflammatory compounds. This study aimed to evaluate the anti-inflammatory effects of Rhus trilobata (Rt) extracts and fractions in lipopolysaccharide (LPS)-induced inflammation models in vitro and in vivo. The aqueous extract (RtAE) and five fractions (F2 to F6) were obtained via C18 solid-phase separation and tested in murine LPS-induced J774.1 macrophages. Key inflammatory markers, such as IL-1β, IL-6, TNF-α, and COX-2 gene expression were measured using RT-qPCR, and PGE2 production was assessed via HPLC-DAD. The in vivo effects were tested in an LPS-induced paw edema model in Wistar rats. Results showed that RtAE at 15 μg/mL significantly decreased IL-1β and IL-6 gene expression in vitro. Fraction F6 further reduced IL-1β, TNF-α, and IL-6 gene expression, COX-2 expression, and PGE2 production. In vivo, F6 significantly reduced LPS-induced paw edema, inflammatory infiltration, and IL-1β and COX-2 protein expression. Chemical characterization of F6 by UPLC/MS-QTOF revealed at least eight compounds with anti-inflammatory activity. These findings support the anti-inflammatory potential of RtAE and F6, reinforcing the medicinal use of Rt.

Assessment of Piper longum L. (Piperaceae) leaves toxicity on the adults of Tribolium castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae)

BackgroundNumerous insect pests attack stored grains causing both qualitative and quantitative losses. The most damaging pest that infests dry stored produce is the red flour beetle, Tribolium castaneum, a secondary pest of stored goods. This pest, especially in its adult stage, exhibits resistance to chemical insecticides, thereby rendering the traditional pesticides ineffective in controlling it. Phyto-derivatives, which are strong insecticides and also ecologically benign, have gained interest as non-chemical solutions for controlling this pest. Hence, the objective of this study was to investigate the potential of Piper longum leaf extract insecticidal action as an environmentally benign insecticide for the first time against the adults of T. castaneum. In this study, P. longum leaf ethanol extract was tested against the adults of T. castaneum by petri dish bioassay method. Ad hoc studies to verify significant mortality for the initial confirmation of adulticidal activity were conducted for 24 h at different dosages of 62.5, 125, 250, 500 and 1000 mg/L of P. longum leaf ethanol extract. Thereafter, dosages set at 10, 20, 30 and 40 mg/L for the fractions of P. longum leaf ethanol extract were conducted. Prior to this, the leaf extract of this plant was subjected to column chromatography for fractionation. The fractions tested for adulticidal activity were subjected to gas chromatography–mass spectroscopy.ResultsSignificant adulticidal action with 100% adult mortality was observed in ethanol extract of P. longum leaves. Among the fourteen fractions (F0–F13) obtained tested, only fractions, F5, F10 and F13, demonstrated adulticidal activity, and the remaining fractions displayed poor activity. One hundred per cent morality was noted in T. castaneum adults after 96 h at 40 mg/L in F5 and F10, and in F13 at 20 mg/L, and their respective LD50 values were 17.6, 26.6 and 10.0 mg/L. The fractions F5, F10 and F13 contained fatty acids, viz., hexadecanoic acid, dotriacontane and heptacosane in F5; tetradecanoic acid and nonadecanoic acid in F10; and octadecanoic acid, aspartame and tridecanoic acid in F13, revealed through gas chromatography–mass spectroscopy.ConclusionsThe results of the study showed that P. longum ethanol leaf extract revealed significant adulticidal activity and is a promising toxic agent to the adults of T. castaneum. The fatty acids in the ethanolic leaf extract fractions of P. longum could have caused toxicity to the adults of T. castaneum. According to the current literature survey, this is the first research report on the adulticidal activity of P. longum leaf extracts against the adults of T. castaneum.

Evaluation of antimalarial compounds in Prosopis africana stem bark fractions against Plasmodium berghei

Prosopis africana, a traditional medicinal plant for malaria treatment, requires further scientific evidence. This study aimed to assess the antimalarial potential of Prosopis africana stem bark fractions in Plasmodium berghei-infected mice and identify the responsible compounds. The aqueous extract underwent partitioning with n-hexane, ethyl acetate, and methanol, followed by antimalarial evaluation. F5 was the most potent fraction based on its antimalarial potential. The study pooled 36 chromatographic separation sub-fractions into five using TLC analysis Rf values. In Plasmodium berghei-infected mice, fraction F5 at 6.25, 12.5, 25, and 50 mg/kg body weight (bw) was tested for antimalarial activity. Compound analysis of F5 utilized ultra-performance liquid chromatography-quadrupolar mass spectrometry. Statistical analysis employed Analysis of Variance and Duncan's multiple range tests (p < 0.05). The methanol, n-hexane, and ethylacetate extracts displayed suppression of P. berghei by 90%, 72.2%, and 73.6%, respectively. Methanol, n-hexane, and ethylacetate extracts suppressed P. berghei by 90%, 72.2%, and 73.6% respectively. F5 showed significantly higher chemo-suppression (67.21%) than F1 (63.94%), F2 (6.99%), F3 (2.81%), and F4 (37.3%). At 12.5 mg/kg bw, F5 suppressed P. berghei by 91.09%, surpassing chloroquine (87.79%), and restored altered hematological indices and CD4+ counts. Compounds identified in F5 included caffeate, linalool, terpinene-4-ol, quinovic acid, quercetin, prosopinine, catechin, and apigeninin. In conclusion, the fraction F5 of P. africana stem bark exhibited higher antimalarial activity than chloroquine and the identified compound might likely contribute to its activity.

High-Throughput Screening System Evaluation of Andrographis paniculata (Burm.f.) Extracts and Their Fractions against Mosquito Vectors.

Infectious diseases that cause illness and/or death in humans can be contracted from mosquito bites. A viable and alternate method of personal protection that can lower the danger of human exposure to mosquito-borne diseases is the use of plant-based repellents. Using a high-throughput screening system, the current work examined the toxicity, contact irritancy, and spatial repellency of Andrographis paniculata crude extract and its fractions against Aedes aegypti, Anopheles minimus, and An. dirus. Five fractions (i.e., F1, F2, F3, F4, and F5) were separated from the crude extract by column and thin layer chromatography and analyzed using high-performance liquid chromatography and mass spectrometry. The major active compounds identified from F3 and F5 were 4-deoxy-11,12-didehydroandrographolide and andrographolide. Three concentrations (1.0, 2.5, and 5.0%) for each of the crude extracts and the five fractions were individually impregnated on nylon netting strips and evaluated against the three mosquito species. Results showed that the highest contact irritancy was elicited by the crude extract at 5% concentration against Ae. aegypti (43.70% escaped). Results of the spatial activity index (SAI) showed that fractions F3 and F5 at 2.5% demonstrated the strongest repellency against Ae. aegypti (SAI = 0.84) and An. minimus (SAI = 0.83), respectively. Both the crude extract and its components did not cause any knockdown or mortality. These findings suggest that fractionation of A. paniculata extracts is valuable in assessing their spatial repellent efficacy against mosquitoes. Fractions F3 and F5 hold promise as natural mosquito repellents and could contribute to developing effective mosquito control strategies.

Development of sustainable Lignin-Based coatings for layered double Hydroxides: Enhancing synergistic Anti-Aging properties in bitumen

Layered Double Hydroxides (LDHs) have poor dispersibility in bitumen and limited effectiveness in enhancing its thermal oxidative aging resistance. To address these issues, this study aims to purify lignin to improve its radical scavenging efficiency and use the purified lignin to coat LDHs (Φ@LDHs), thereby increasing their lipophilicity and radical scavenging efficiency for use in bitumen. sodium lignosulfonate (SL) was fractionated into five parts using a stepwise water/ethanol fractionation method, and these fractions were then applied to coat the LDHs to produce Φ@LDHs. The morphology, antioxidant activity, and chemical structure of Φ@LDHs were analyzed. Subsequently, the effect of Φ@LDHs on the aging properties of bitumen was evaluated. The test results indicated that SL and its fractions effectively coated the surfaces of LDHs, thereby enhancing their interlayer spacing and hydrophobicity. Among all fractions, lower molecular weight fractions, F4 and F5, retained most of the phenolic hydroxyl groups in SL, demonstrating significant antioxidant activities. The incorporation of F4@LDHs and F5@LDHs effectively mitigated changes in rheological properties of bitumen and oxidation product formation during thermal oxidation and UV aging, with F5@LDHs showing the best performance. Furthermore, a synergistic anti-aging mechanism was elucidated for the F4@LDHs and F5@LDHs composites. The purified F4 and F5 fractions not only improved the compatibility of LDHs with bitumen but also delayed the oxidative reactions initiated by free radicals to achieve a synergistic anti-aging effect.

Application of INTDOSKIT tool for assessment of uncertainties on dose coefficients for ingestion of uranium by workers

The International Commission on Radiological Protection (ICRP) has developed the Human Alimentary Tract Model (HATM) to calculate radiation doses from the ingestion of radionuclides for the protection of workers and the public. In parallel, the ICRP's Occupational Intake of Radionuclides (OIR) series provides biokinetic models and dose coefficients based on a reference human, primarily for regulatory purposes. Although these coefficients are not usually checked for uncertainties, the investigation of such uncertainties is crucial to ensure their reliability in radiation protection. This study uses INTDOSKIT, a software tool developed using the R programming language and RStudio as the Integrated Development Environment (IDE), to calculate doses and explore uncertainties following ingestion of U-238 by workers. Two scenarios were investigated: Case I, using the latest HATM model and the systemic uranium model from ICRP publications 100 and 137 respectively, and Case II, based on data from literature, using older ICRP models. In both cases, intake of U-238 in its soluble form (Type F) was modeled and the results were validated using the ICRP OIR dataviewer software. Following validation, uncertainty and sensitivity analyses were performed. In Case I, uncertainties were assigned to the particle transport parameters in both the systemic model model and HATM as well as the uranium uptake fraction into the blood. While in Case II they were limited to the uptake fraction (f1) and the systemic uranium model. A Monte Carlo simulation of 60,000 runs was performed for both cases, sampling model parameters from their respective probability distributions to generate dose distributions. The influence of each parameter on these distributions was also analyzed. Probability distributions were inferred to the calculated dose values using the maximum likelihood estimation method and the Kolmogorov-Smirnov goodness-of-fit. The results showed that for both cases the committed effective dose coefficient, e (50), followed a lognormal distribution. Case I had a geometric mean of 3.2E-08 Sv/Bq and GSD of 2.0, while case II had a slightly lower geometric mean of 3.1E-08 Sv/Bq and GSD of 1.9. Sensitivity analysis showed that the main contributor to dose uncertainty was the fraction of uranium absorbed from the small intestine into the blood. Both cases showed similar trends, with slightly higher results in case I. Overall, this study demonstrates the effectiveness of INTDOSKIT in calculating dose coefficients and analyzing uncertainties. It suggests that while the ICRP reference values remain useful for protection, the incorporation of additional statistical measures and distribution characteristics could further enhance radiation protection strategies.

Bioassay guided isolation and compounds identification of the anti-diabetic fractions of (rosemary) Rosmarinus officinalis leaves extract

BackgroundDiabetes mellitus is a metabolic disease characterized by prolonged elevated blood glucose levels. It is a common health problem with a high mortality and morbidity to the human race. A number of medicinal plants such as rosemary (Rosmarinus officinalis) have been used for the treatment of diabetes. Most of the anti-diabetic conventional drugs have been found to have some side effects and there is therefore need to explore new sources of anti-diabetic drugs. The aim of the current study was to investigate the possibility of getting anti-diabetic compounds from R. officinalis that can be used as leads for drug discovery.MethodologyR. officinalis leaves were macerated in 50% methanol in dichloromethane and the crude extract fractionated by column chromatography. The obtained fractions were subjected to an in-vitro alpha-amylase inhibition assay. The anti-hyperglycemic potential of the fractions was evaluated in diabetic induced Wistar rats. The most potent fractions were analyzed by gas chromatography-mass spectrophotometry (GC-MS) for identification of the compounds.ResultsA total of 21 chromatographic fractions were assembled with different alpha- amylase inhibition activity. Eleven of the fractions had more than 30% alpha-amylase inhibition activity. The ethyl acetate fraction had the highest inhibition potential (LC50 of 2.8 μg/mL). The anti-diabetic assay in rats showed that fractions (F1) and (F4) had highest blood glucose reduction of 44.5 ± 0.4 and 52.8 ± 1.3%, respectively (p < 0.05). GC-MS analysis of fractions F1 and F4 showed the presence of 21 and 23 compounds in F1 and 23, respectively.ConclusionThis study has demonstrated that R. officinalis crude extract fractions obtained from 50% methanol in dichloromethane possesses alpha-amylase inhibitory and anti-hyperglycemic activities as well as secondary metabolites with varying chemical structures. The hexane and hexane/ethyl acetate (8/2) fractions showed most potent alpha-amylase inhibition with high anti-hyperglycemic activity giving hope of a possibility of obtaining lead compounds for new anti-diabetic drugs.

Exploring the anticandidal potential: Evaluating leaf extract efficacy and GC–MS metabolite profiling of Chenopodium album var. album L.

IntroductionChenopodium album var. album L., renowned as lamb's quarters, is a versatile medicinal plant, revealing its efficacy in treating various ailments. This study evaluated the in vitro antifungal activity of ethyl acetate extract of C. album var. album L. MethodsChenopodium album leaf extract demonstrated antifungal potential via the Agar well-diffusion and Broth-dilution method. Active fractions, obtained through column chromatography, were rigorously tested against Candida spp. The most potent fraction underwent detailed analysis with High-Performance Thin-Layer Chromatography (HPTLC), Gas Chromatography Mass Spectrometry (GC–MS), and Fourier-Transform Infrared Spectroscopy (FT-IR), for comprehensive characterization. ResultsThe ethyl acetate extract (EAE) exhibited significant antifungal activity, with Candida albicans being the most susceptible (inhibition zone of 14.67 ± 0.58a). Fractionation of the EAE yielded a highly active F2 fraction, which showed a remarkable zone of inhibition against C. albicans (17.50 ± 1.00a). Comprehensive examinations, including HPTLC, GC–MS, and FT-IR, identified 12 compounds in the F2 fraction, and the major compounds were Furo [2,3-c] pyridine, 2,3-dihydro-2,7-dimethyl, Bis (2-ethylhexyl) phthalate, and 2,3,4,4-Tretrapropyl-1-(trimethylsilyl)-1-(trimethylsilyloxy)-1,3-diaza-2,4-diborabutane. This study marks the first report of these 12 compounds identified from the F2 fraction of C. album. ConclusionThe present study highlights the antifungal activity of EAE and its fractions from C. album var. album that could be further explored for the selection of leads that may contribute to novel drug development.

Indole alkaloids from Ochreinauclea maingayi (Rubiaceae) as butyrylcholinesterase inhibitors and their paralysis effect in transgenic Caenorhabditis elegans

This study investigated the butyrylcholinesterase (BChE) inhibitory activity of harmane (1), naucledine (2), and dihydrodeglycocadambine (3) isolated from fractions F7 and F9 of Ochreinauclea maingayi. Both fractions demonstrated significant inhibition, exceeding 80%, against BChE at 100 µg/mL. Compound 2, is the most potent inhibitor, exhibiting an IC50 value of 22.08 µM, followed by 1 and 3 (IC50 23.96 and 30.32 µM, respectively). Docking studies revealed that 1 and 2 effectively bind to BChE, with binding energies of −51.24 and −57.17 kcal/mol, respectively. Kinetic analysis of 2 indicated mixed-mode inhibition of BChE, with a Ki of 6.08 μM. In the paralysis assay, 1 showed a weak delay in paralysis and reduced the paralysis ratio from 72.59 ± 4.7% to 60.00 ± 7.0% (12.59% reduction) followed by 2 with 70.00 ± 1.7% (2.59% reduction) compared with negative standard (DMSO 0.1%) on human amyloid β-protein in a transgenic Caenorhabditis elegans (CL4176) model.

Activity concentration of indoor radon and its short-lived progeny (218Po, 214Pb, and 214Po) and their effect on atmospheric ionization rate in Sana'a, Yemen

Exposure to ionizing radiation inside houses, especially radionuclides of radon and its progeny, poses serious health risks that can be exacerbated when inhaled as a result of interaction with human lung tissue. Also, air ionization is mainly due to these radionuclides. Therefore, accurate measurements of radon activity concentrations and its short-lived progeny are required to assess dose and environmental pollution and estimate ionization rates in indoor environments. For this purpose, we employed a previously tested and approved reliable method, following the three-count procedure. This method is based on airborne radon progeny sampling on polycarbonate membrane filters and alpha counting using a passive α-dosimetry technique with CR-39 detectors. The method also relies on a PC-based software we developed for solving mathematical equations and calculating all the necessary physical quantities. In this study, the concentrations of radon and individual short-lived radon progeny were measured in 20 houses in Sana'a, Yemen. Measurement conditions and meteorological variables were considered. The average activity concentrations of 222Rn, Equilibrium-Equivalent Concentration (EEC), 218Po, 214 Pb, and 214Po were 73.1 ± 6.0, 29.2 ± 2.4, 44.4 ± 3.6, 30.5 ± 2.5, and 23.2 ± 1.9 Bq.m-3, respectively. The calculated average unattached fractions f1(218Po), f2(214 Pb), and fp were found to be 0.24, 0.04, and 0.07 % respectively. The annual average values of ion-pair production rate caused by 222Rn and their progeny and air ion concentration, were 27.25 ions.cm-3s-1 and 1829 ions.cm-3 respectively. The annual effective dose was estimated to be 1.93 ± 0.16 mSv.y−1, well lower than the recommended 10 mSv.y−1.

Transformation of As and Cd associated with Fe-Mn-modified biochar during simultaneous remediation on the contaminated soil.

Here, Fe- and Mn-modified biochar (BC-Fe-Mn) was applied to simultaneously stabilize As and Cd in the contaminated soil. The removal efficiencies for NaHCO3-extractable As and DTPA-extractable Cd by BC-Fe-Mn were 60.8% and 49.6%, respectively. The speciation analyses showed that the transformation to low-crystallinity Fe-bound (F3) As, Fe-Mn oxide-bound (OX) of Cd, and residual As and Cd was primarily attributed to stabilizing the two metal(loid)s. Moreover, the correlation analyses showed that the increase of As in F3 fraction was significantly and positively associated with the increase of OX fraction Mn (r = 0.64). Similarly, OX fraction Cd was increased notably with increasing OX fraction Fe (r = 0.91) and OX fraction Mn (r = 0.76). In addition, a novel dialysis experiment was performed to separate the reacted BC-Fe-Mn from the soil for intensively investigating the stabilization mechanisms for As and Cd by BC-Fe-Mn. The characteristic crystalline compounds of (Fe0.67Mn0.33)OOH and Fe2O3 on the surface of BC-Fe-Mn were revealed by SEM-EDS and XRD. And FTIR analyses showed that α-FeOOH, R-COOFe/Mn+, and O-H on BC-Fe-Mn potentially served as the reaction sites for As and Cd. A crystalline compound of MnAsO4 was found in the soil treated by BC-Fe-Mn in the dialysis experiment. Thus, our results are beneficial to deeper understand the mechanisms of simultaneous stabilization of As and Cd by BC-Fe-Mn in soil and support the application of the materials on a large scale.

Unveiling the anti-Candida albicans activity of bioactive fractions from mining soil-dwelling Streptomyces sp. DS104

Due to an increase in the incidence of Candida albicans infection and its growing resistance to currently used antifungal drugs, further development of more potent antimicrobials is needed. This study sought to assess the antifungal properties of Streptomyces sp. DS104, isolated from a mining soil, against C. albicans and identify the bioactive substances that are produced by this strain. Ethyl acetate supernatant's crude extract showed a high inhibition against C. albicans, with a minimal inhibitory concentration value as low as 4 µg/mL. Two bioactive fractions (F5 and F6) have been isolated using silica gel column chromatography, demonstrating zones of inhibition against C. albicans measuring 28.33 ± 01.52 to 29.00 ± 02.64 mm. The GC–MS analysis was carried out on the purified fractions and allowed the identification of 21 compounds predominantly consisting of carboxylic acids and pyrrolopyrazine derivatives. In Fraction F5, the major compounds detected are pentadecanoic and hexadecanoic acids, constituting the highest area percentages of 41.99 % and 33.77 %, respectively. As for fraction F6, the predominant compounds include pyrrolo[1,2-a]pyrazine-1,4‑dione, hexahydro-3-(2-methylpropyl) and pyrrolo[1,2-a]pyrazine-1,4‑dione, hexahydro-3-(methylphenyl), accounting for area percentages of 20.18 % and 12.16 %, respectively. Various other bioactive compounds, such as isocoumarins, and phenolic compounds, had previously been identified as components with antifungal activity, were also detected. It's worth highlighting that three compounds specifically, 2-hydrazino-8‑hydroxy-4-phenylquinoline, hydroxyphenylacetic acid, and 3-carboxaldehyde-1H-indole have been reported for the first time to be detected within the Streptomyces genus. These results suggested that Streptomyces sp. DS104 is a promising strain and thus appears to be a potential candidate for the development of antifungal drugs.

Prodelphinidins enhance dentin matrix properties and promote adhesion to methacrylate resin

ObjectiveInvestigate the bioactivity and stability of Rhodiola rosea (RR) fractions as a natural source of prodelphinidin gallate (PDg) on dentin collagen via analysis of the viscoelastic and resin-dentin adhesive properties of the dentin matrix. MethodsThe biomimicry and stability of RR subfractions (F1, F2, F3 and F4) with collagen were determined by dynamic mechanical analysis (DMA). DMA used a strain sweep method to assess the dentin matrix viscoelastic properties [storage (E’), loss (E”), and complex (E*) moduli and tan δ] after treatment, 7-, 30- and 90-days of storage in simulated body fluids (SBF). Resin-dentin interface properties were assessed after 1 and 90-days in SBF by microtensile bond strength test and confocal laser scanning microscopy. Data were analyzed using two and one-way ANOVA and post-hoc tests (α = 0.05). ResultsRR fractions increased dentin matrix complex (96 - 69 MPa) and storage (95 - 68 MPa) moduli, compared to the control (∼9 MPa) in the ranking order: F2 ≥ F3 = F1 = F4 > control (p < 0.001). Treatment did not affect tan δ values. After 30- and 90-days, RR-treated dentin E*, E’ and tan δ decreased (p < 0.001). F2 fraction yielded the highest microtensile bond strength (43.9 MPa), compared to F1, F4 (35.9 - 31.7 MPa), and control (29 MPa). RR-treated interfaces mediated stable surface modifications and enhanced collagen-methacrylate resin interactions at the bioadhesive interface. SignificanceProdelphinidin gallates from RR are potent and reasonably stable biomimetic agents to dentin. Higher potency of F2 fraction with the dentin matrix and the adhesive interface is associated with a degree of polymerization of 2–3 and gallo(yl) motifs.

Angiotensin-I-converting enzyme and renin inhibitions by antioxidant shrimp shell protein hydrolysate and ultrafiltration peptide fractions

Protein hydrolysates with 20% and 30% degrees of hydrolysis (DH) were produced from alcalase hydrolysis of Pacific white shrimp shells. The study aimed to investigate the antioxidant and antihypertensive activities of these protein hydrolysates, focusing on peptide fractions of various molecular weights (MW) obtained through ultrafiltration. The 30% DH protein hydrolysate (SSPH) showed the highest antioxidant activities and was subjected to ultrafiltration to produce peptides of various molecular weights (MW) (F1:10–50, F2:5–10, F3:3–5, F4:1-3, and F5:<1 kDa). SSPH showed higher hydrophilic and non-essential amino acid content, among which alanine and aspartic acid were at higher levels. The obtained peptide fractions were analyzed for various antioxidant activities and inhibitory activities against angiotensin-converting enzyme and renin. F4 and F2 samples showed the highest DPPH (2.00 mM TE/g sample) and ABTS (60.68 mM TE/g sample) radical scavenging activities, respectively. Superoxide and hydrogen peroxide radical scavenging activity augmented with decreasing MW, aside from for F1, which showed higher activity than F2–F4 (p < 0.05). The highest ACE-inhibitory activity was found for the F2 fraction (44%–55%), and renin inhibitory activity increased with decreasing MW, in which F5 (12%–22%) was the most effective (p < 0.05). F5 showed 6 major peptides, among which KLVRGSKS and LPKKCLSARPG were the dominant sequences. Thus, this study suggests that shrimp shell protein isolate could be an alternative protein source for peptides with various antioxidant and antihypertensive activities, with potential applications in functional foods and nutraceuticals aimed at mitigating oxidative stress and hypertension.

Isolation of flavonoids from Tagetes patula(French Marigold) flowers: Cytotoxic and oxidant activity in human cervical carcinoma cells

Tagetes patula, known as French Marigold, belongs to the family Asteraceae. Human papillomavirus infection is considered one of the causes of cervical cancer. This study assessedthe cytotoxic activity and intracellular oxidative capacity of compounds isolated from extract of T. patulaflowers as anti-cancer cervical agents. Fraction F6 of n-butanol extract was subjected to column chromatography and HPLC-ESI-MS. The isolated compounds of T. patulawere used to examine cytotoxic activity and the production of total reactive oxygen species in SiHa and HeLa cells; the cells were also characterized using scanning electron microscopy. Patulitrin was cytotoxic to SiHa and HeLa cells. An increase in ROS production was observed at different times of treatment of cells with patuletin and patulitrin. Scanning electron microscopy showed morphological changes in SiHa and HeLa cells. Thus, compounds isolated from T. patulahave great treatment potential against cervical cancer.

Withanolides production by the endophytic fungus Penicillium oxalicum associated with Withania somnifera (L.) Dunal.

Withanolides are steroidal lactones with diverse bioactive potential and their production from plant sources varies with genotype, age, culture conditions, and geographical region. Endophytic fungi serve as an alternative source to produce withanolides, like their host plant, Withania somnifera (L.) Dunal. The present study aimed to isolate endophytic fungi capable of producing withanolides, characterization and investigation of biological activities of these molecules. The methanolic fungal crude extract of one of the fungal isolates WSE16 showed maximum withanolide production (219mg/L). The fungal isolate WSE16 was identified as Penicillium oxalicum based on its morphological and internal transcribed spacer (ITS) sequence analysis and submitted in NCBI (accession number OR888725). The methanolic crude extract of P. oxalicum was further purified by column chromatography, and collected fractions were assessed for the presence of withanolides. Fractions F3 and F4 showed a higher content of withanolides (51.8 and 59.1mg/L, respectively) than other fractions. Fractions F3 and F4 exhibited antibacterial activity against Staphylococcus aureus with an IC50 of 23.52 and 17.39µg/ml, respectively. These fractions also showed antioxidant activity (DPPH assay with IC50 of 39.42 and 38.71µg/ml, superoxide anion scavenging assay with IC50 of 41.10 and 38.84µg/ml, and reducing power assay with IC50 of 42.61 and 41.40µg/ml, respectively) and acetylcholinesterase inhibitory activity (IC50 of 30.34 and 22.05µg/ml, respectively). The withanolides present in fraction 3 and fraction 4 were identified as (20S, 22R)-1a-Acetoxy-27-hydroxywitha-5, 24-dienolide-3b-(O-b-D-glucopyranoside) and withanamide A, respectively, using UV, FTIR, HRMS, and NMR analysis. These results suggest that P. oxalicum, an endophytic fungus isolated from W. somnifera, is a potential source for producing bioactive withanolides.

Lemon basil seed-derived peptide: Hydrolysis, purification, and its role as a pancreatic lipase inhibitor that reduces adipogenesis by downregulating SREBP-1c and PPAR-γ in 3T3-L1 adipocytes.

The purpose of this study is to assess the bioactive peptides derived from the defatted lemon basil seeds hydrolysate (DLSH) for their ability to inhibit pancreatic lipase, decrease intracellular lipid accumulation, and reduce adipogenesis. Response surface methodology (RSM) was employed to optimize trypsin hydrolysis conditions for maximizing lipase inhibitory activity (LI). A hydrolysis time of 387.06 min, a temperature of 49.03°C, and an enzyme concentration of 1.61% w/v, resulted in the highest LI with an IC50 of 368.07 μg/mL. The ultrafiltration of the protein hydrolysate revealed that the fraction below 0.65kDa exhibited the greatest LI potential. Further purification via RP-HPLC identified the Gly-Arg-Ser-Pro-Asp-Thr-His-Ser-Gly (GRSPDTHSG) peptide in the HPLC fraction F1 using mass spectrometry. The peptide was synthesized and demonstrated LI with an IC50 of 0.255 mM through a non-competitive mechanism, with a constant (Ki) of 0.61 mM. Docking studies revealed its binding site with the pancreatic lipase-colipase complex. Additionally, GRSPDTHSG inhibited lipid accumulation in 3T3-L1 cells in a dose-dependent manner without cytotoxic effects. Western blot analysis indicated downregulation of PPAR-γ and SREBP-1c levels under GRSPDTHSG treatment, while an increase in AMPK-α phosphorylation was observed, suggesting a role in regulating cellular lipid metabolism. Overall, GRSPDTHSG demonstrates potential in attenuating lipid absorption and adipogenesis, suggesting a prospective application in functional foods and nutraceuticals.

Exploring novel peptides in adzuki bean and mung bean hydrolysates with potent antibacterial activity

SummaryThis study delved into the antimicrobial potential of hydrolysates from adzuki bean and mung bean protein concentrates, created using Alcalase and Flavourzyme enzymes. Targeting common foodborne pathogens (Salmonella Typhimurium, Salmonella Enteritidis, Vibrio parahaemolyticus, Staphylococcus aureus, and Bacillus subtilis) and Escherichia coli, the research underscored the crucial role of peptidase in generating antibacterial peptides. Fractions F2 of Flavourzyme hydrolysed adzuki bean (ABF) and F4 of Flavourzyme hydrolysed mung bean (MBF) showed the highest antibacterial activity against S. Typhimurium and S. aureus, respectively, whereas F2 of Alcalase hydrolysed mung bean (MBA) exhibited very poor activity against S. aureus. TripleTOF LC–MS/MS analysis revealed small cationic antibacterial peptides, with lysine being prominent against S. aureus and proline–proline against S. Typhimurium. These findings highlighted the significant contribution of protease enzymes in fostering antibacterial peptide production, offering valuable insights for developing effective antimicrobial agents in the context of food safety.