Crude Extract Research Articles

Nematicidal activity of Pleurotus djamor metabolites on Meloidogyne incognita under greenhouse conditions

This study evaluated the nematicidal activity of the cell-free crude extract (CFCE) of Pleurotus djamor on Meloidogyne incognita under greenhouse conditions. The fungus was grown in a solid medium for 14 days in the dark. Distilled water was added to the medium, filtered, and centrifuged, and the proteolytic activity of the CFCE produced was measured. Three experimental groups were established: control group (distilled water), CFCE (denatured enzymes), and CFCE (active enzymes). The nematicidal activity of the extracts was assessed in vitro on second-stage juveniles (J2) and eggs of M. incognita, separately. Under greenhouse conditions, CFCEs were applied to tomato seedlings grown on substrate infested with M. incognita eggs. About the in vitro tests, the active CFCE showed a significant difference concerning the control group (p<0.05) according to the t-test, with a 22% reduction in eggs. On the other hand, about J2, both treatments showed a significant difference compared to the control (p<0.01) with a 100% reduction, but no significant difference between themselves (p>0.05). Similarly, the treatments in the greenhouse showed the same statistical behavior as the control (p<0.01). However, there was no significant difference between them (p>0.05) regarding the number of eggs and the number of galls. The reduction in the number of eggs was 94 and 97 % and in the number of galls was 71 and 67% for denatured and active CFCE, respectively. This study points to the nematicidal action of metabolites on J2 and the activity of enzymes on M. incognita eggs.

Nematicidal efficacy of Cymbopogon nardus, Pelargonium graveolens essential oils and Annona squamosa extract against Meloidogyne incognita and M. graminicola

A crude hexane extract of seed kernel was processed into fatty acid methyl esters (FAME) using methanolic-sulfuric acid. Herbal mixtures (seed extracts of A. squamosa and essential oils of Cymbopogon nardus (citronella) and Pelargonium graveolens (geranium)) were evaluated against root knot nematode (Meloidogyne spp). The mortality of J2s of M. incognita in hexane and methanol extracts (AHE+AME) of A. squamosa with geranium oil (GO) ranged from 50.75-89.75% to 71.50-99.00% at 31.25 to 1000 μgmL-1 in 24 to 96 hours. The number of galls after CO+AHE+AME treatment was 4.0 ± 0.0 and 5.67 ± 0.58 galls/seedling and were recorded at 1000 and 500 μgmL-1 treatments on 30 days after inoculation, compared to carbofuran (3.0 ± 0.0 and 4.3 ± 0.6 galls/seedling) at 1000 and 500 μgmL-1 and velum prime (3.6 ± 0.6 and 4.6 ± 0.6 galls/seedling) at 1000 and 500 μgmL-1. The maximum shoot length of brinjal seedlings was recorded in CO+AHE+AME treatment compared to carbofuran (47.3 ± 0.6cm) and velum prime (48.7 ± 0.6 cm) at 1000 μg mL-1 in soil drenching application. Methanol and hexane extracts (AHE+AME) obtained from Annona squamosa seeds were investigated for their nematicidal properties. M. graminicola in direct-seeded rice at the nursery level and M. incognita in brinjal (eggplant) were controlled for up to sixty days when treated with these mixtures.

Serodiagnosis of urogenital schistosomiasis and profiling of immunoreactive protein(s) in Schistosoma haematobium soluble egg and adult worm antigens.

To achieve schistosomiasis eradication plan by 2030, the development of efficient diagnosis is crucial. This study focuses on assessing the immunodiagnostic potential of S. haematobium (Sh) soluble egg antigen (SEA) and worm antigen (SWA) for urogenital schistosomiasis. Urine microscopy identified 50 S. haematobium-positive and 50 negative samples from a total of 500 examined. An additional 50 samples from a non-endemic area were included, bringing the total number of samples used for the assay to 150. Indirect ELISA immunoassays using SEA and SWA as the probing antigens evaluated 50 sera samples each from Sh positive, negative endemic (NE), and non-endemic (NNE) individuals. SDS-PAGE analysis of crude protein extracts was conducted, followed by Western blot analysis using primary antibodies from pooled Sh-infected sera samples. Diagnostic performance was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), sensitivity, and specificity. The AUC values for Sh SEA and SWA were 0.75 and 0.76 in NE samples, and 0.91 and 0.89 in NNE samples, respectively. Sensitivities 90 (95% CI: 78.64 - 95.65)/ 64.71 (95% CI: 52.17 - 75.92), and specificities 50 (95% CI: 36.64 - 63.36)/ 81.25 (95% CI: 63.56 - 92.79) were recorded for SEA and SWA, respectively in NE samples. In addition, sensitivities 90 (78.64 - 95.65)/ 92 (95% CI: 80.77 - 97.78), and specificities 72 (95% CI: 58.33 - 82.53)/ 72.00 (95% CI: 57.51 - 83.77) were recorded for SEA and SWA, respectively in NNE samples. The mean antibody titer against Sh SEA in infected samples was significantly higher than in non-infected samples (P <0.0001). Eight (8) immunoreactive protein bands; 4 each of SEA and SWA were identified, indicating potential for diagnostic tool development. Sh SEA and SWA demonstrate promise for diagnosing urogenital schistosomiasis in both endemic and non-endemic regions.

In vivo evaluation of anti-diarrheal activity of hydroalcoholic extract and solvent fractions of the leaf of Leucas deflexa Hook.f (Lamiaceae) in rodents

The emergence of antibiotic-resistant bacteria has intensified the search for novel antidiarrheal drug. Plant-derived extracts offer promising alternatives due to their ability to modulate gut motility and enhance water absorption. The leaves of Leucas deflexa Hook.f., a plant native to Ethiopia, have been traditionally utilized in the treatment of diarrheal diseases. Nonetheless, its traditional application has yet to be scientifically confirmed. Hence, the aim of the present study was to evaluate antidiarrheal activity of the crude extract and fractions of the leaf of Leucas deflexa Hook.f (Lamiaceae). Healthy Swiss albino mice were randomly divided into five groups per experimental model: castor oil-induced diarrhea, gastrointestinal motility, and anti-enteropooling assays. The control group received distilled water (10ml/kg), while the positive control was treated with loperamide (3mg/kg). The remaining groups were administered various dosages (LD100, LD200, and LD400 mg/kg) of a hydroalcoholic extract or its fractions from Leucas deflexa Hook.f. The crude extract and chloroform fraction substantially (P<0.001) delayed the onset of diarrhea, reduced fecal frequency, and decreased total fecal weight at all administered doses compared to the negative control. The butanol and aqueous fractions exhibited a substantial antidiarrheal effect at doses of 200mg/kg and 400mg/kg, respectively. All treatments, including the crude extract, chloroform, butanol, and aqueous fractions, substantially (P<0.001) reduced the volume and weight of intestinal contents at various doses. Additionally, the chloroform, aqueous fraction, and crude extract at doses of 100, 200, and 400mg/kg substantially inhibited gastrointestinal motility compared to the negative control. The results of this study corroborate the traditional use of Leucas deflexa leaves in the treatment of diarrhea. These findings give a scientific justification for further exploration of the underlying mechanisms and potential therapeutic applications of this plant extract.

Mitigating aluminum chloride-induced toxicity in Drosophila melanogaster with peptide fractions from Euphorbia species

This study assessed the antioxidative and protective effects of peptide extracts from selected Euphorbia species on Aluminum Chloride (AlCl3)-induced toxicity in Drosophila melanogaster. Euphorbia humifusa (EHU), Euphorbia hirta (EHI), and Euphorbia graminae (EHG) were screened for bioactive peptides. The crude peptide extract and partially purified peptide fractions of all the plants were subjected to preliminary antioxidants activities through 2, 2-diphenyl-1-picrylhyhdrazyl (DPPH), and nitric oxide (NO) scavenging activities. The most active peptide fraction was subjected to biochemical studies using Drosophila melanogaster. Flies were treated with AlCl3 (100 mg/kg diet), peptide fraction (5 and 10 mg/kg diet), and cotreatment of AlCl3 and the fraction, respectively. After treatment, flies were homogenized for the determination of total thiol and Glutathione (non-protein thiol) content, catalase and Glutathione-S-transferase activities, and nitric oxide (nitrite/nitrate) and hydroperoxide levels. The antioxidant screening revealed that the peptide fraction from Euphorbia humifusa (PEHU) was the most significant compared to the control (ascorbic acid). The PEHU (5 and 10 mg/kg diet) maintained the redox status of the flies in the biochemical study. The PEHU significantly counteracted AlCl3-induced reduction in antioxidants (catalase, GST, GSH and Total thiol), increased nitric oxide levels, and acetylcholinesterase activity and prevented behavioral deficits flies. Hence, the peptide fraction of Euphorbia humifusa may shield against the life-threatening effects of free radicals associated with aluminum chloride toxicity.

Investigate the efficacy of compounds found in the crude extract of the fungus Aspergillus niger on seeds germination of zucchini

For a long time, fungi have been considered the richest source of bioactive metabolites with numerous applications. Hence, this study was performed to identify various bioactive compounds contained in the ethyl acetate extract of the fungus Aspergillus niger that isolates from infected garlic by using Gas Chromatography-Mass Spectrometry and their effect on seed germination of Zucchini. The current results showed a variety of bioactive compounds such as n-hexadecanoic acid, Hexadecanoic acid, Methyl stearate, Benzoic acid, Hexamethyl-1,3,5-trithiane, and cis-aconitic anhydride with an area percentage of 15.37, 13.93, 6.87, 5.67, 4.75 and 3.59% respectively. The results were performed using the fungus extract that protected the zucchini seed from pathogenic fungi (Botrytis cinerea, Fusarium oxysporum, and Rhizoctonia solani) when soaking for less than 72 h and promoted increased seed germination by high percentage ratio of seeding (66.67%, 80.00%, and 80.00%) respectively, radical length (66.67, 80.00 and 7.67), plumule length (4.67, 7.67 and 6.00) and Number of secondary roots (7.00,8.00 and 7.00) respectively compared with control treatment.

Phytochemical and volatile components evaluation of antimicrobial root extracts of &lt;i&gt;Dichrostachys cinerea&lt;/i&gt; (sickle bush)(&lt;i&gt;Fabaceae&lt;/i&gt;) (L) Wight &amp; Arn

The crude ethanolic extract of the root of Dichrostachys cinerea (Fabaceae) was screened for phytochemicals and antimicrobial activity against Staphylococcus aureus, Escherichia coli, Pseudomonas aureginosa, Salmonella typhi, and Candida albicans using agar-well diffusion method. The phytochemical screening of the root extract results showed the presence of alkaloids, flavonoids, essential oils, phenols, terpenes, tannins, unsaturated sterols, and saponins. However, coumarins, and anthocyanins were absent. The crude extract was subjected to column chromatographic fractionation to give nine fractions of which two, DCR1 and DCR2, were subjected to gas chromatography-mass spectrometry (GC-MS) analysis. The crude extract and column chromatographic fractions DCR1 and DCR2 gave MIC of 1 mg/mL against Staphylococcus aureus, but were not active against the other test organisms. The gas chromatography-mass spectrometry analysis of the antibacterial fractions, DCR1 and DCR2, based on standard MS library data revealed the presence of caryophellene, phthalates, a phenol and fatty acids and their ester derivatives. These phytoconstituents could account for the biological properties of the extracts of the plant. The results of this work therefore support its use in ethno-medicinal preparations, particularly against infections.

Antiprotozoal Natural Products from Endophytic Fungi Associated with Cacao and Coffee

Background: Collectively, leishmaniasis and Chagas disease cause approximately 8 million cases and more than 40,000 deaths annually, mostly in tropical and subtropical regions. The current drugs used to treat these diseases have limitations and many undesirable side effects; hence, new drugs with better clinical profiles are needed. Fungal endophytes associated with plants are known to produce a wide array of bioactive secondary metabolites, including antiprotozoal compounds. In this study, we analyzed endophytic fungal isolates associated with Theobroma cacao and Coffea arabica crop plants, which yielded extracts with antitrypanosomatid activity. Methods: Crude extracts were subjected to bioassay-guided isolation by HPLC, followed by spectrometric and spectroscopic analyses via mass spectrometry (MS) and nuclear magnetic resonance (NMR), Results: Compounds 1–9 were isolated and displayed novel antitrypanosomal and antileishmanial activities ranging from 0.92 to 32 μM. Tandem liquid chromatography–mass spectrometry (LC–MS) analysis of the organic extracts from different strains via the feature-based Global Natural Products Social (GNPS) molecular networking platform allowed us to dereplicate a series of metabolites (10–23) in the extracts. Molecular docking simulations of the active compounds, using the 3-mercaptopyruvate sulfurtransferase protein from L. donovani (Ld3MST) and the cruzipain enzyme from T. cruzi as putative molecular targets, allowed us to suggest possible mechanisms for the action of these compounds. Conclusions: The isolation of these antiprotozoal compounds confirms that crop plants like coffee and cacao harbor populations of endophytes with biomedical potential that confer added value to these crops.

Antimicrobial activity of blue mold (Penicillium italicum) Filtrates against some species of pathogenic bacteria

Background: Penicillium saprophytic species, which primarily consume organic biodegradable materials, is a common example of a fungal species. The preparation filtrates of Penicillium italicum, a saprophyte on citrus fruits frequently linked to post-harvest diseases in this crop, were included in the current investigation. Objective: This investigation aimed determine the most efficient culture medium for the production of antibacterial secondary metabolites. Patients and Methods: included identifying the growth medium for P. italicum and produce its metabolites through the use of gas chromatography-mass spectrometry (GC MS) for both solid-state fermentation filtrate (SSFF) and liquid fermentation filtrate (LFF). Additionally, the antimicrobial activity of the mold filtrate against certain pathogenic bacteria was assessed using the agar well diffusion method. Results: indicated that the biomass used for mold growth was heavier in SSFF than LFF, and according to the findings, the selective active isolate's crude filtrate from two duplicates of the P. italicum Yeast Extract Sucrose YES culture medium was 0.063 mg, while the crude extract from rotten orange (as a solid medium) containing P. italicum was 0.11 mg. Tetracosane and other substances with a track record of therapeutic activity were found in the two mold extracts, according to GC MS data. Overall, both SSFF and LFF demonstrated antibacterial activity against Klebsiella pneumonia, Escherichia coli, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus aureus, with the inhibition zones± standard deviation (IZ± SD) being 24.7± 0.57, 18.2± 0.28, 26.3± 0.59, 21.6± 0.51, and 32.8± 0.21 (for SSFF) mm and 0.0, 12.3±0.57, 28.16±0.20, 19.3±1.15, and 28±0.2 (for LFF) mm, respectively. Conclusion: the filtrate of P. italicum from a natural medium (rotted orange) as a solid state fermentation was more weighted and gave many effective metabolites compared to what was produced by liquid fermentation on a synthetic medium, and both liquid and solid fermentation filtrates demonstrated efficacy against harmful bacteria.

Toxigenic effects of sponges and benthic diatoms on marine invertebrates and their possible biotechnological applications

Secondary metabolites play important physiological roles being bioactive as defences against other organisms, or attractive signals used for various purposes, including reproduction. Their production and the emission in the environment may be viewed as an adaptive feature subjected to evolutionary selection. They were demonstrated to be useful for applications in various biotechnological fields, such as pharmaceutical, nutraceutical and cosmeceutical. Sponges and microalgae, including diatoms, are the most promising sources of bioactive compounds from the sea. We aimed at detecting the ecotoxicological effects of crude extracts and fractions obtained from three marine sponges, Geodia cydonium, Haliclona (Halichoclona) vansoesti and Agelas oroides and two benthic diatoms, Nanofrustulum shiloi and Cylindrotheca closterium on model marine organisms. We tested their effects on the Mediterranean purple sea urchin, Paracentrotus lividus, and on two diatoms, Phaeodactylum tricornutum and Cylindrotheca closterium, chosen because they are considered standard indicators for assessment of ecological impacts. Our results showed that extracts and fractions from both sponges and diatoms may be harmful for model invertebrates. However, eggs appeared “protected” from sponge allelochemicals when still unfertilized. The majority of sponge fractions exhibited noticeable impacts during the post-fertilization treatments. In contrast, fractions from diatoms notably increased the rate of malformations compared to the control, both in pre- and post-fertilization treatments.

Onion Peel Extract Prevents Intestinal Inflammation via AMK-Activated Protein Kinase Activation in Caco-2/HT-29 Cells.

Obesogenic diets cause intestinal inflammation and dysfunction. Polyphenols have shown a positive impact on reducing inflammation in in vitro studies. However, their bioactivity may not be the same in the in vivo system due to structural alteration by the gastrointestinal digestive process. The purpose of this study was to investigate the anti-inflammatory effect of onion peel and its major bioactive compound, quercetin, in the intestine and further examine the impact of intestinal digestion on this effect. Onion peel extract (OPE) and quercetin (Q) were digested using gastrointestinal digestive enzymes in vitro and then treated into lipopolysaccharide (LPS)-stimulated Caco-2/HT-29 cells. Genes and proteins related to tight junction, inflammation, and epithelial integrity were measured. OPE and digested OPE (DOPE) had a higher protective effect on LPS-induced tight junction and inflammatory genes and paracellular permeability than Q and digested Q (DQ). DOPE was more effective than OPE, while digestion did not change the activity of Q. The anti-inflammatory effect of OPE and Q with or without digestion was achieved by inhibiting nuclear factor kappa B through AMP-activated protein kinase-activated silent mating-type information regulation 2 homolog 1. It was the first to find that a crude extract, after undergoing gastrointestinal digestion, demonstrated a notably superior anti-inflammatory effect in the cell study, suggesting the consumption of onion peels could potentially yield similar benefits in the human intestine. This discovery underscores the potential of onion peel polyphenols in combating intestinal inflammation, making them a compelling area of research for future therapeutic applications using food byproducts.

Antioxidant and anti-glycation activities of Mandevilla velutina extract and effect on parasitemia levels in Trypanosoma cruzi experimental infection: In vivo, in vitro and in silico approaches

Ethnopharmacological relevanceMandevilla velutina (Mart. Ex Stadelm.) Woodson, known in Brazil as "infalível" and "jalapa", is a medicinal plant native from the Cerrado region (Brazilian Savannah). The underground organ (xylopodium) of this species is prepared as ethanolic extract or infusion and it is commonly used in traditional medicine to treat snake venom. Although, locals and indigenous populations from Cerrado have used M. velutina for the treatment of infection by Trypanosoma cruzi (Chagas’ disease). Aim of the studyThis study aimed to evaluate the in vitro antioxidant and anti-glycation activities of the crude hydroethanolic extract of M. velutina xylopodium. Besides, it aimed to evaluate its effect on parasitemia levels in vivo T. cruzi experimental infection. In addition, this study aimed to determine possible interactions between the main compound of the extract and molecular targets associated with survival and virulence of T. cruzi in silico approaches. Materials and methodsDetermination of total polyphenols, flavonoids and steroidal aglycones content were performed. In addition, high performance liquid chromatography (HPLC) was carried out to identify main compounds of the extract. Antioxidant activity was determined by DPPH radical scavenging, ferric ion reducing power (FRAP), Thiobarbituric acid reactive species (TBARS) and Oxygen Radical Absorbance Capacity (ORAC) methods. Anti-glycation activity was demonstrated through relative mobility in electrophoresis (RME), determination of free amino groups and inhibition of AGEs formation. Determination of the action of extract in parasitemia levels was performed by T. cruzi experimental infection of mice and nitrite levels were measured in the serum of animals evaluated in this study. Molecular docking analyses of the main compound (Velutinol A) with DNA and molecular targets associated with survival and virulence of T. cruzi. ResultsPhytoconstituents evaluation exhibited the presence polyphenols, flavonoids and steroidal aglycone, and HPLC identified the major presence of Velutinol A. Antioxidant and anti-glycation evaluations showed that the extract present significant activity in all methods evaluated. In addition, extract reduced the number of trypomastigotes and increased the survival of treated animals. The treatment using extract showed an interference in the synthesis of physiological nitric oxide as an immune response to infection. In silico assays demonstrated interaction between Velutinol A and DNA and molecular targets of T. cruzi. ConclusionsThe results showed that the hydroethanolic extract of M. velutina xylopodium contains bioactive compounds including polyphenols, flavonoids and steroidal aglycones (mainly Velutinol A) of which may be responsible for the antioxidant, anti-glycation and anti-parasitic activity against T. cruzi. Trypanocidal activity of M. velutina compounds may be linked to their influence on NO synthesis during infection and/or their capacity to bind and inhibit molecules associated to virulence and survival of T. cruzi.

Ethanol crude leaf extract of weeping fig (Ficus benjamina) causes mild hepatotoxicity in Sprague dawley rats

Weeping fig (Ficus benjaminaeaf extract has diverse medicinal properties but little is reported about its hepatotoxicity. This study determined the mean lethal dose (LD50) and investigated the effects of F. benjamina ethanol crude leaf extract on biochemical parameters and liver histology of Sprague Dawley rats. Twenty-nineemale rats weighing 133-204 g were used. The LD50 was determined with nine rats based on Lorke’s method. The experimental groups consisted of twenty rats, divided into four groups of five. Each group received treatment as follows: Control (feed and water only) and low, medium, and high doses (500, 1000, 1500 mg/kg respectively) of the extract orally for 21 days. All animals were weighed and sacrificed using Ketamine intra-peritoneal injection. Blood samples were collected for biochemical parameters of total bilirubin, conjugated bilirubin, aspartate aminotransferase (AST) alanine aminotransferase (ALT), and alkaline phosphatase (ALP). Liver tissues were removed and processed by the formalin-fixed-paraffin wax-embedding method. The tissue blocks were sectioned and subjected to Hematoxylin/Eosin and Masson trichrome staining. The extract’s LD50 was &gt;5000 mg/kg. The rats’ body weights did not change statistically (p=0.985). Total bilirubin (p=0.003), conjugated bilirubin, AST, ALT, and ALP values (p=0.001) increased significantly. The AST high-doseroup had the highest fold increase (4.8). The liver histology showed mild sinusoidal dilation at 500 mg/kg. There was marked hemorrhage and fibrosis at medium and high doses. Although the extract had relatively low acute toxicity, 1000 and 1500 mg/kg doses were associated with mild hepatotoxicity characterized by veno-occlusion disease. The 500 mg/kg dose is safer for medicinal purposes.

Thermothelomyces thermophilus cultivated with residues from the fruit pulp industry: enzyme immobilization on ionic supports of a crude cocktail with enhanced production of lichenase.

β-Glucans comprise a group of β-D-glucose polysaccharides (glucans) that occur naturally in the cell walls of bacteria, fungi, and cereals. Its degradation is catalyzed by β-glucanases, enzymes that catalyze the breakdown of β-glucan into cello-oligosaccharides and glucose. These enzymes are classified as endo-glucanases, exo-glucanases, and glucosidases according to their mechanism of action, being the lichenases (β-1,3;1,4-glucanases, EC 3.2.1.73) one of them. Hence, we aimed to enhance lichenase production by Thermothelomyces thermophilus through the application of response surface methodology, using tamarind (Tamarindus indica) and jatoba (Hymenaea courbaril) seeds as carbon sources. The crude extract was immobilized, with a focus on improving lichenase activity, using various ionic supports, including MANAE (monoamine-N-aminoethyl), DEAE (diethylaminoethyl)-cellulose, CM (carboxymethyl)-cellulose, and PEI (polyethyleneimine)-agarose. Regarding lichenase, the optimal conditions yielding the highest activity were determined as 1.5% tamarind seeds, cultivation at 50°C under static conditions for 72h. Moreover, transitioning from Erlenmeyer flasks to a bioreactor proved pivotal, resulting in a 2.21-fold increase in activity. Biochemical characterization revealed an optimum temperature of 50°C and pH of 6.5. However, sustained stability at varying pH and temperature levels was challenging, underscoring the necessity of immobilizing lichenase on ionic supports. Notably, CM-cellulose emerged as the most effective immobilization medium, exhibiting an activity of 1.01 U/g of the derivative (enzyme plus support), marking a substantial enhancement. This study marks the first lichenase immobilization on these chemical supports in existing literature.

Magnetic protein aggregates generated by supramolecular assembly of ferritin cages - a modular strategy for the immobilization of enzymes.

Efficient and cost-effective immobilization methods are crucial for advancing the utilization of enzymes in industrial biocatalysis. To this end, in vivo immobilization methods relying on the completely biological production of immobilizates represent an interesting alternative to conventional carrier-based immobilization methods. This study aimed to introduce a novel immobilization strategy using in vivo-produced magnetic protein aggregates (MPAs). MPA production was achieved by expressing gene fusions of the yellow fluorescent protein variant citrine and ferritin variants, including a magnetically enhanced Escherichia coli ferritin mutant. Cellular production of the gene fusions allows supramolecular assembly of the fusion proteins in vivo, driven by citrine-dependent dimerization of ferritin cages. Magnetic properties were confirmed using neodymium magnets. A bait/prey strategy was used to attach alcohol dehydrogenase (ADH) to the MPAs, creating catalytically active MPAs (CatMPAs). These CatMPAs were purified via magnetic columns or centrifugation. The fusion of the mutant E. coli ferritin to citrine yielded fluorescent, insoluble protein aggregates, which are released upon cell lysis and coalesce into MPAs. MPAs display magnetic properties, as verified by their attraction to neodymium magnets. We further show that these fully in vivo-produced protein aggregates can be magnetically purified without ex vivo iron loading. Using a bait/prey strategy, MPAs were functionalized by attaching alcohol dehydrogenase post-translationally, creating catalytically active magnetic protein aggregates (CatMPAs). These CatMPAs were easily purified from crude extracts via centrifugation or magnetic columns and showed enhanced stability. This study presents a modular strategy for the in vivo production of MPAs as scaffold for enzyme immobilization. The approach eliminates the need for traditional, expensive carriers and simplifies the purification process by leveraging the insoluble nature and the magnetic properties of the aggregates, opening up the potential for novel, streamlined applications in biocatalysis.

Allelopathic Effects of Vachellia nubica, Vachellia tortilis and Hyphaene compressa on Germination and Seedling Growth of Prosopis juliflora

Background: Prosopis juliflora (Sw.) DC is an invasive species negatively impacting plant ecology and livelihoods in introduced areas. Native tree species may produce allelochemicals that can inhibit growth of invasive species, resulting in a natural management technique. Aim: This study aimed to determine allelopathic effect of three native tree species (V. tortilis, V. nubica, and H. compressa) on seed germination and early seedling growth of P. juliflora to identify the tree species that can be used to suppress P. juliflora invasion as part of its management strategy. Methodology: Crude Leaf and root extracts of the native trees at 0%, 1%, 2.5%, 5%, and 10% concentrations were prepared. P. juliflora seeds were subjected to 24 crude extract treatments and a control in a completely randomized design in Petri dishes. The seeds were uniformly moistened with 10 ml extracts daily in a germination chamber for 14 days, and germination counts were done after every 2 days. Shoot and root length and wet weight were measured after 14 days. The data were subjected to three-way ANOVA and means separated by Tukey. Results: Prosopis seed coats treated with V. tortilis and V. nubica leaf extracts turned darkish before germination while germination of seeds in 10% leaf extracts of V. nubica aborted. Seed germination percentage in 10% leaf extracts of V. tortilis (30%) and V. nubica (23%) were the lowest compared to control (90%). P. juliflora seedlings under 5% V. nubica and 10% V. tortilis died before the experiment period. Depending on crude extract concentrations, percent inhibition on germination characteristics were; percent germination (21-74%), wet weight (6-49%), plumule length (10-47%), and radicle length (7-86%) of germinating P. juliflora. Conclusion: The native tree species studied have allelopathic effects on the germination of P. juliflora and thus can be promoted to suppress invasiveness of P. juliflora.

Strategies to reduce the culture medium costs for a high‐yield and high‐selectivity bio‐based 2,3‐butanediol production

AbstractBio‐based 2,3‐butanediol (2,3‐BDO) production on a large scale depends on critical factors, such as culture medium, oxygen supply, pH and biosafety. In this study, three strategies for reducing culture medium costs were investigated: carbon/nitrogen (C/N) ratio; low‐cost nitrogen sources (crude yeast extract, brewer's yeast extract, corn steep liquor, urea, sodium nitrate, ammonium chloride, ammonium sulfate and dibasic ammonium phosphate); and microbial pH autoregulation. Batch fermentations were performed in a microaerobic environment using wild‐type and safe Paenibacillus peoriae NRRL BD 62. The yield and selectivity of 2,3‐BDO were used as control variables. A ratio between 2,3‐BDO production and glucose consumption (YP/S) of almost 80% and an optical purity of 87% levo‐2,3‐BDO, with no acetoin accumulation, were achieved in an NH4Cl‐based medium at C/N = 8.5 g/g and without external pH control, considering an initial glucose of 10 g/L. Based on Free on Board prices, a 63% savings in culture medium costs was achieved by replacing commercial yeast extract at pH 5. Validation assays with higher initial glucose concentrations showed a YP/S of 0.40 g/g and an optical levo‐/meso‐2,3‐BDO ratio of 1:0.8, with negligible acetoin accumulation. Therefore, the NH4Cl‐based medium at C/N = 8.5 g/g and without pH control was considered economically promising for high‐yield and high‐selectivity bio‐based 2,3‐BDO production.

Brine shrimp lethality and antioxidant property of &lt;i&gt;Lagenaria breviflora&lt;/i&gt; (Benth.) Roberty fruit crude extract and fractions

Background: Oxidative injury plays a pivotal impact in the development of human diseases, and it is a major component of the pathophysiology of several inflammation-linked human medical disorders. Objective: The focus of this study was to evaluate the cytotoxicity and antioxidant potential of the fruit of Lagenaria breviflora (Benth.) Roberty (LB). Methods: The antioxidant capacity of L. breviflora's crude and solvent fractions was assessed using standard techniques by measuring the total phenolic content, total flavonoid content, and DPPH radical scavenging activity. The data we0re analyzed using One-way ANOVA, and Dunnett's Multiple Comparison was used to establish the threshold. Results: The total flavonoid contents of fruit methanol extract, LB hexane, chloroform, and ethyl acetate fractions were 32.00 ± 0.94, 224.32 ±0.05, 224.06 ±0.14, and 2615.67 ± 13.20 mg gallic acid equivalent/g of extract, respectively. In contrast, the total phenolic contents were 5218.83 ± 3.18, 2615.67 ± 13.20, and 4553.00 ± 24.27 mg gallic acid equivalent/g of extract, respectively. The crude extract of LB, hexane fraction, chloroform, and ethyl acetate fraction displayed IC50 values of 242.57 ± 8.90, 262.91 ± 6.49, 96.17 ± 2.74, and 221.45 ± 0.83 μg/mL, respectively, for their ability to scavenge DPPH radicals. Ascorbic acid and rutin had IC50 values of 2.76 ± 0.01 and 20.6 ± 9.26 μg/mL, respectively. Conclusions: Fruit from L. breviflora (LB) exhibited strong antioxidant potential, which may have resulted from the fruit's phenolic and flavonoid content. This demonstrates the reason this herb is used in traditional medicine. Keywords: Oxidative stress, brine shrimp lethality assay, total phenolic contents, total flavonoid contents, DPPH, Lagenaria breviflora fruit.

In Vitro Antitumor, Antioxidant, and Hemolytic Activities of Chlorella sorokiniana Methanol Extracts and Collective Fractions

Chlorella species are fast-growing microalgae with significant industrial applications. The aim of the present study was to investigate the antitumor, antioxidant, and hemolytic activities of Chlorella sorokiniana UTEX 1230 crude methanol extracts and fractions. Ch. sorokiniana crude methanol extracts and collective fractions (CFs) were obtained from lyophilized biomass by maceration and column chromatography. Antitumor assays against murine lymphoma L5178Y-R and human breast cancer MCF-7 cells were performed by the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction technique, using human peripheral blood mononuclear cells (PBMC) as the control group. Antioxidant and hemolytic activities were evaluated using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay (DPPH) and erythrocyte hemolysis, respectively. We showed that crude methanol extracts (IC50) increased L5178Y-R and MCF-7 cell growth inhibition, without affecting PBMC. In addition, all evaluated CFs showed significantly higher antioxidant activity than the positive control (ascorbic acid). CF3 and CF4 showed the highest cytotoxicity against L5178Y-R, whereas CF3, CF4, and CF5 caused the highest antitumor activity against MCF-7 cells. CF3, CF4, and CF5 induced significantly higher hemolytic activity compared with all other fractions. CF characterization revealed loliolide, cinnamic acid, methyl dihydrojasmonate, salsalvamide A, 1-monolinolenin, cryptophycin 29, costunolide, riboflavin lumicrome, and germicidin B, which have been related to antitumor and antioxidant activities. In conclusion, we demonstrated that Ch. sorokiniana extracts and fractions possess antitumor and antioxidant potential, without affecting human erythrocytes and PBMC.

Phytochemical screening and toxicity assessment of compounds isolated from the leaves of Mangifera indica L. for the control of Spodoptera litura (Lepidoptera; Noctuidae)

This study aimed to analyze the phytochemical composition and antioxidative capabilities of mango leaves (Mangifera indica L., Nam Dok Mai), indigenous to Sa Kaeo Province, Thailand. Various solvents with differing polarities, including n-hexane, DCM, ethyl acetate, and MeOH, were utilized for leaf extraction. The findings revealed the existence of eight groups of phytochemical compounds: alkaloids, flavonoids, coumarins, saponins, tannins, terpenoids, steroids, and cardiac glycosides. The MeOH crude extract exhibited the highest concentration of total phenolic compounds at 409.88 ±0.02 mg GAE/g. Furthermore, the MeOH crude extract demonstrated the strongest antioxidant activity, with an IC50 value of 0.52 ±0.02 μg/ml, as determined by the DPPH method. High-performance liquid chromatography (HPLC) was employed to identify gallic acid and mangiferin in the MeOH crude extract. Laboratory tests were conducted using the topical application method to evaluate the toxicity of the M. indica leaf crude extract on 2nd instar Spodoptera litura larvae. The MeOH crude extract exhibited high efficacy, with an LD50 value of 10.58 µg per larvae within 24 hours. Gallic acid and mangiferin were identified as the primary active ingredients, with LCD50 values of 1.19 µg per larvae and 1.90 µg per larvae, respectively, within 24 hours. Additionally, the impact on detoxification enzymes (24 hours post-treatment) was assessed in surviving 2nd instar S. litura larvae using the topical application method. The MeOH extract treatment resulted in 1.31-fold inhibition of carboxylesterase (CE), 1.31-fold inhibition of glutathione-S-transferase (GST), and 1.32-fold inhibition of acetylcholinesterase (AChE).