scholarly journals Evaluation of the anthelmintic properties of a traditional remedy based on a mixture of red algae using an in vitro assay on gastrointestinal nematodes of donkeys

2021 ◽  
Vol 4 (1) ◽  
pp. 1-7
Author(s):  
Michela Maestrini ◽  
◽  
Marcelo Beltrão Molento ◽  
Simone Mancini ◽  
Francesco Saverio Robustelli della Cuna ◽  
...  
Keyword(s):  
Red Algae ◽  
Gastrointestinal Nematode ◽  
Gastrointestinal Nematodes ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Egg Hatch ◽  
Vitro Assay ◽  
Reference Drug ◽  
Negative Controls

The anthelmintic properties and composition of an Italian traditional anthelmintic remedy based on a red algae mixture (RAE) was assessed using the egg hatch test (EHT). The ability of different dilutions \((1.0, 5.0, 50, or 100%)\) of RAE was determined and compared with the positive and negative controls against gastrointestinal nematode (GIN) of donkeys. The experiment was performed in triplicate. Data were analysed using the ANOVA and Tukey test. In the mixture, Palisada tenerrima, Laurencia intricata and Laurencia spp. red algae were identified. The \(100%\) RAE was able to totally inhibit the egg hatch, showing an efficacy comparable \((P < 0.05)\) to that of the reference drug \((98.7%)\). An egg hatch reduction of \(89.5, 43.7\), and \(23.4%\) was observed at \(50, 5\) and \(1%\) dilutions, respectively. In conclusion, RAE was able to inhibit the egg hatch of GIN of donkeys in a concentration-dependent manner with a correlation coefficient \((R2)\) of \(0.968\), corroborating with its anthelmintic effect.

scholarly journals In Vitro Anthelmintic Activity of Saponins from Medicago spp. Against Sheep Gastrointestinal Nematodes

Molecules ◽  
2020 ◽  
Vol 25 (2) ◽  
pp. 242 ◽  
Author(s):  
Michela Maestrini ◽  
Aldo Tava ◽  
Simone Mancini ◽  
Doriana Tedesco ◽  
Stefania Perrucci
Keyword(s):  
Small Ruminants ◽  
Anthelmintic Activity ◽  
Gastrointestinal Nematodes ◽  
Animal Origin ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Reference Drug ◽  
Synthetic Drugs

Gastrointestinal strongyle nematodes (GIS) are included among the most important parasites of small ruminants. The widespread drug resistance and drug residues in products of animal origin have increased the interest in the search for natural compounds with anthelmintic activity as a valid alternative to current synthetic drugs. The aim of the present investigation was to test the ‘in vitro’ anthelmintic activity of saponins and prosapogenins from different Medicago species, selected for their importance as a forage crop worldwide for animal feeding. From these plants, saponin mixtures were extracted, purified and used at scalar concentrations to evaluate their anthelmintic activities against sheep gastrointestinal strongyles (GISs), by the egg hatch test. Treated and untreated controls were used as the comparison. Data were statistically analyzed, and EC50 and EC90 were also calculated. All saponins and prosapogenins showed inhibiting effects on GIS eggs in a concentration-dependent manner. At higher concentrations, most of them showed an efficacy comparable to the reference drug (Thiabendazole 3 µg/mL) (P < 0.001). With 1.72 mg/mL EC50 and 3.84 mg/mL EC90, saponin from M. polymorpha cultivars Anglona was the most active. Obtained results encourage further studies aimed at evaluating the efficacy ‘in vivo’ of saponins which resulted as most effective ‘in vitro’ in this study.

scholarly journals In vitro Anthelmintic Activity of Two Aloe-derived Active Principles against Sheep Gastrointestinal Nematodes

2017 ◽  
Vol 12 (12) ◽  
pp. 1934578X1701201
Author(s):  
Gianluca Fichi ◽  
Matteo Mattellini ◽  
Elisa Meloni ◽  
Guido Flamini ◽  
Stefania Perrucci
Keyword(s):  
Larval Development ◽  
Larval Mortality ◽  
Plant Secondary Metabolites ◽  
Anthelmintic Activity ◽  
Gastrointestinal Nematodes ◽  
Egg Hatch ◽  
Reference Drug ◽  
Faecal Samples ◽  
Aloe Emodin

The in vitro anthelmintic activity on sheep gastrointestinal strongyle (GIS) eggs and larvae of 0.5% aloin and 0.1% aloe-emodin was investigated. From fresh faecal samples collected by ewes naturally infected by Haemonchus, Trichostrongylus and Teladorsagia nematodes, GIS eggs were isolated and cultivated in Petri dishes (100 eggs/dish). For the in vitro evaluation of the anthelmintic activity of tested compounds, the Egg hatch test (EHT), the Larval development test (LDT) and the Larval mortality/paralysis test (LMT) were used. In each assay, the activity of tested compounds was compared to untreated and treated (0.1% thiabendazole, TBZ) controls. Six repetitions were made through the experiment. Obtained data were statistically elaborated using the X2 test. In EHT, 0.5% aloin gave highly significantly different (P<0.01) results from the untreated controls. In LDT, both 0.1% aloe-emodin and 0.5% aloin almost completely prevented the larval development from L1 to L3, showing no significant differences (P<0.01) when compared to TBZ. In LMT, larval mortality observed in 0.5% aloin treated plates was significantly higher (P<0.01) than that observed in TBZ treated controls. These results show the in vitro anthelmintic properties on sheep GIS of the examined plant secondary metabolites. In LDT and/or LMT, the activity of 0.5% aloin and 0.1% aloe-emodin was comparable to or higher than that of the reference drug.

scholarly journals In Vitro Anthelmintic Activity of Saponins Derived from Medicago spp. Plants against Donkey Gastrointestinal Nematodes

2019 ◽  
Vol 6 (2) ◽  
pp. 35 ◽  
Author(s):  
Michela Maestrini ◽  
Aldo Tava ◽  
Simone Mancini ◽  
Federica Salari ◽  
Stefania Perrucci
Keyword(s):  
Anthelmintic Activity ◽  
Gastrointestinal Nematodes ◽  
Deionized Water ◽  
Chemical Compositions ◽  
Egg Hatching ◽  
Egg Hatch ◽  
Reference Drug ◽  
Fecal Samples ◽  
Nematode Eggs

With the aim to find new effective natural compounds for the control of nematodes, the in vitro anthelminthic properties of purified 1% saponins showing different chemical compositions and derived from Medicago sativa (MS), Medicago arborea (MA), Medicago polymorpha cultivar ‘Santiago’ (MPS), M. polymorpha cultivar ‘Anglona’ (MPA), and 1% prosapogenins from M. sativa (MSp), were evaluated and compared. As a source of nematode eggs, pooled fresh fecal samples taken from dairy donkeys naturally infected by gastrointestinal nematodes were used. From fecal samples, eggs were recovered, suspended in deionized water, and used immediately in the bioassay (egg hatch test). The activity of the tested compounds was compared to positive (0.1% thiabendazole) and negative (deionized water and 1% DMSO) controls. All experiments were repeated in triplicate and the obtained data were statistically analyzed. All the tested plant compounds caused a significant (p < 0.05) inhibition of nematode egg hatching (>80%). Moreover, all saponins and prosapogenins showed in vitro anthelmintic properties statistically comparable to that of the reference drug (p < 0.05), except for MPS extract. Obtained results showed that the different Medicago saponins evaluated in this study possess high anthelmintic properties against gastrointestinal nematodes of dairy donkeys, although to a different extent depending on their composition.

Early mouse endoderm is patterned by soluble factors from adjacent germ layers

Development ◽  
2000 ◽  
Vol 127 (8) ◽  
pp. 1563-1572 ◽  
Author(s):  
J.M. Wells ◽  
D.A. Melton
Keyword(s):  
Primitive Streak ◽  
In Vitro Assay ◽  
Specific Gene ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Germ Layers ◽  
Vitro Assay ◽  
Organ Specific ◽  
Early Mouse

Endoderm that forms the respiratory and digestive tracts is a sheet of approximately 500–1000 cells around the distal cup of an E7.5 mouse embryo. Within 2 days, endoderm folds into a primitive gut tube from which numerous organs will bud. To characterize the signals involved in the developmental specification of this early endoderm, we have employed an in vitro assay using germ layer explants and show that adjacent germ layers provide soluble, temporally specific signals that induce organ-specific gene expression in endoderm. Furthermore, we show that FGF4 expressed in primitive streak-mesoderm can induce the differentiation of endoderm in a concentration-dependent manner. We conclude that the differentiation of gastrulation-stage endoderm is directed by adjacent mesoderm and ectoderm, one of the earliest reported patterning events in formation of the vertebrate gut tube.

scholarly journals In Vitro Modulation of Glibenclamide Transport by P-glycoprotein Inhibitory Antidiabetic African Plant Extracts

Planta Medica ◽  
2019 ◽  
Vol 85 (11/12) ◽  
pp. 987-996
Author(s):  
Udoamaka F. Ezuruike ◽  
Elisabetta Chieli ◽  
Jose M. Prieto
Keyword(s):  
Transport Model ◽  
Diabetic Patients ◽  
Diabetes Incidence ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Reference Drug ◽  
P Glycoprotein ◽  
Efflux Ratio ◽  
Syzygium Guineense

AbstractThe rise of diabetes incidence in Nigeria enhances the use of popular remedies that may interact with conventional therapies. The aqueous extracts of 27 popular Nigerian “antidiabetic” plants were tested for their in vitro effects on glutathione levels within HepG2 cells, P-glycoprotein (P-gp)-mediated Rh-123 efflux activity in Caco-2 vincristine-resistant cells, and modulation of glibenclamide transport in Caco-2 monolayers. The extract from Ximenia americana significantly depleted intracellular glutathione at 100 µg/mL similarly to the reference buthionine sulphoximine (p < 0.05). Other 10 extracts raised glutathione levels. Eight extracts inhibiting P-gp efflux in a concentration-dependent manner (p < 0.01) were selected for further evaluation in a bi-directional transport model across Caco-2 monolayers: Annona senegalensis, Bridellia ferruginea, Cassytha filiformis, Daniellia ogea, Khaya ivorensis, Syzygium guineense, Terminalia avicennioides, and X. americana. When interferences in paracellular transport were discarded, only 3 of them may be modulating the efflux ratio of glibenclamide (efflux ratio: 2.65 ± 0.13) in the same manner the reference drug verapamil (efflux ratio: 1.14 ± 0.25, p < 0.01) does: Syzygium guineense (efflux ratio: 1.70 ± 0.23, p < 0.01), Terminalia avicennioides (efflux ratio: 1.80 ± 0.25, p < 0.05), and X. americana (efflux ratio: 1.66 ± 0.10, p < 0.01). HPLC-UV analyses for P-gp inhibitors in these extracts revealed several phenolic compounds such as rutin, gallic acid, and ellagic acid reported to decrease P-gp expression and/or directly modify its function. In conclusion, some popular herbal medicines used by Nigerian diabetic patients are here shown to potentially affect glibenclamide absorption at concentrations that could be reached in the intestinal tract.

scholarly journals Homotypic Lysosome Fusion in Macrophages: Analysis Using an In Vitro Assay

1997 ◽  
Vol 139 (3) ◽  
pp. 665-673 ◽  
Author(s):  
Diane M. Ward ◽  
Jonathan D. Leslie ◽  
Jerry Kaplan
Keyword(s):  
Fusion Reaction ◽  
In Vitro Assay ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Dependent Manner ◽  
Toxin A ◽  
Concentration Dependent Manner ◽  
Vitro Assay ◽  
Sensitive Factor

Lysosomes are dynamic structures capable of fusing with endosomes as well as other lysosomes. We examined the biochemical requirements for homotypic lysosome fusion in vitro using lysosomes obtained from rabbit alveolar macrophages or the cultured macrophage-like cell line, J774E. The in vitro assay measures the formation of a biotinylated HRP–avidin conjugate, in which biotinylated HRP and avidin were accumulated in lysosomes by receptor-mediated endocytosis. We determined that lysosome fusion in vitro was time- and temperature-dependent and required ATP and an N-ethylmaleimide (NEM)-sensitive factor from cytosol. The NEM-sensitive factor was NSF as purified recombinant NSF could completely replace cytosol in the fusion assay whereas a dominant-negative mutant NSF inhibited fusion. Fusion in vitro was extensive; up to 30% of purified macrophage lysosomes were capable of self-fusion. Addition of GTPγs to the in vitro assay inhibited fusion in a concentration-dependent manner. Purified GDP-dissociation inhibitor inhibited homotypic lysosome fusion suggesting the involvement of rabs. Fusion was also inhibited by the heterotrimeric G protein activator mastoparan, but not by its inactive analogue Mas-17. Pertussis toxin, a Gαi activator, inhibited in vitro lysosome fusion whereas cholera toxin, a Gαs activator did not inhibit the fusion reaction. Addition of agents that either promoted or disrupted microtubule function had little effect on either the extent or rate of lysosome fusion. The high value of homotypic fusion was supported by in vivo experiments examining lysosome fusion in heterokaryons formed between cells containing fluorescently labeled lysosomes. In both macrophages and J774E cells, almost complete mixing of the lysosome labels was observed within 1–3 h of UV sendai-mediated cell fusion. These studies provide a model system for identifying the components required for lysosome fusion.

Chickpea (Cicer arietinum L.) Lectin Exhibit Inhibition of ACE-I, α-amylase and α-glucosidase Activity

2019 ◽  
Vol 26 (7) ◽  
pp. 494-501 ◽  
Author(s):  
Sameer Suresh Bhagyawant ◽  
Dakshita Tanaji Narvekar ◽  
Neha Gupta ◽  
Amita Bhadkaria ◽  
Ajay Kumar Gautam ◽  
...  
Keyword(s):  
Biological Effects ◽  
Exchange Chromatography ◽  
Health Concern ◽  
Carbohydrate Binding ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Ic50 Values ◽  
Chickpea Lectin

Background: Diabetes and hypertension are the major health concern and alleged to be of epidemic proportions. This has made it a numero uno subject at various levels of investigation. Glucosidase inhibitor provides the reasonable option in treatment of Diabetes Mellitus (DM) as it specifically targets post prandial hyperglycemia. The Angiotensin Converting Enzyme (ACE) plays an important role in hypertension. Therefore, inhibition of ACE in treatment of elevated blood pressure attracts special interest of the scientific community. Chickpea is a food legume and seeds contain carbohydrate binding protein- a lectin. Some of the biological properties of this lectin hitherto been elucidated. Methods: Purified by ion exchange chromatography, chickpea lectin was tested for its in vitro antioxidant, ACE-I inhibitory and anti-diabetic characteristic. Results: Lectin shows a characteristic improvement over the synthetic drugs like acarbose (oral anti-diabetic drug) and captopril (standard antihypertensive drug) when, their IC50 values are compared. Lectin significantly inhibited α-glucosidase and α-amylase in a concentration dependent manner with IC50 values of 85.41 ± 1.21 ҝg/ml and 65.05 ± 1.2 µg/ml compared to acarbose having IC50 70.20 ± 0.47 value of µg/ml and 50.52 ± 1.01 µg/ml respectively. β-Carotene bleaching assay showed antioxidant activity of lectin (72.3%) to be as active as Butylated Hydroxylanisole (BHA). In addition, lectin demonstrated inhibition against ACE-I with IC50 value of 57.43 ± 1.20 µg/ml compared to captopril. Conclusion: Lectin demonstrated its antioxidant character, ACE-I inhibition and significantly inhibitory for α-glucosidase and α-amylase seems to qualify as an anti-hyperglycemic therapeutic molecule. The biological effects of chickpea lectin display potential for reducing the parameters of medically debilitating conditions. These characteristics however needs to be established under in vivo systems too viz. animals through to humans.

Improved Method for Obtaining of Arctigenin from Arctium Lappa L. and its Antiproliferative Effect on Human Hepatocarcinoma HepG2 Cells

2020 ◽  
Vol 16 (3) ◽  
pp. 358-362
Author(s):  
Renan S. Teixeira ◽  
Paulo H.D. Carvalho ◽  
Jair A.K. Aguiar ◽  
Valquíria P. Medeiros ◽  
Ademar A. Da Silva Filho ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Crude Extract ◽  
Hepg2 Cells ◽  
Liver Carcinoma ◽  
Adhesion Assay ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Arctium Lappa ◽  
Nih 3T3

Background: Arctigenin is a lignan found in Arctium lappa L. (Asteraceae) that displays anti-inflammatory activities. Previous studies showed that the crude extract of A. Lappa has antitumor activity in human liver carcinoma, lung and stomach cancer cells. The aim of this study was to obtain arctigenin from A. lappa L., as well as to evaluate its antiproliferative effects in cells of liver carcinoma (HepG2) and fibroblasts (NIH/3T3). Methods: Arctigenin was obtained from the hydrolysis of arctiin, which was isolated from the crude extract of A. lappa. The effects of arctigenin and arctiin on HepG2 cell viability and cell adhesion were analyzed by MTT method. Adhesion assay was also carried out to evaluate the antitumor activity. Results: Our results showed that the analytical process to obtain arctigenin was fast and easy. In vitro experiments showed that arctigenin (107-269 μM) decreased HepG2 cells viability and did not cause cytotoxicity on NIH/3T3 cells. Arctigenin (27-269 μM) demonstrated anti-adhesion in HepG2 cells in a concentration-dependent manner, when compared with control. Conclusion: These results suggest a promising pharmacological activity for arctigenin as an antiproliferative compound.

scholarly journals Human Neuronal Cell Lines as An In Vitro Toxicological Tool for the Evaluation of Novel Psychoactive Substances

2021 ◽  
Vol 22 (13) ◽  
pp. 6785
Author(s):  
Valeria Sogos ◽  
Paola Caria ◽  
Clara Porcedda ◽  
Rafaela Mostallino ◽  
Franca Piras ◽  
...  
Keyword(s):  
Cell Death ◽  
Neuronal Cell ◽  
In Vitro Study ◽  
Dependent Manner ◽  
Novel Psychoactive Substances ◽  
Psychoactive Substances ◽  
Concentration Dependent Manner ◽  
Mechanisms Of Toxicity ◽  
Neuronal Cell Lines

Novel psychoactive substances (NPS) are synthetic substances belonging to diverse groups, designed to mimic the effects of scheduled drugs, resulting in altered toxicity and potency. Up to now, information available on the pharmacology and toxicology of these new substances is very limited, posing a considerable challenge for prevention and treatment. The present in vitro study investigated the possible mechanisms of toxicity of two emerging NPS (i) 4′-methyl-alpha-pyrrolidinoexanophenone (3,4-MDPHP), a synthetic cathinone, and (ii) 2-chloro-4,5-methylenedioxymethamphetamine (2-Cl-4,5-MDMA), a phenethylamine. In addition, to apply our model to the class of synthetic opioids, we evaluated the toxicity of fentanyl, as a reference compound for this group of frequently abused substances. To this aim, the in vitro toxic effects of these three compounds were evaluated in dopaminergic-differentiated SH-SY5Y cells. Following 24 h of exposure, all compounds induced a loss of viability, and oxidative stress in a concentration-dependent manner. 2-Cl-4,5-MDMA activates apoptotic processes, while 3,4-MDPHP elicits cell death by necrosis. Fentanyl triggers cell death through both mechanisms. Increased expression levels of pro-apoptotic Bax and caspase 3 activity were observed following 2-Cl-4,5-MDMA and fentanyl, but not 3,4-MDPHP exposure, confirming the different modes of cell death.

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