Investigating in Vivo and in Vitro Effects of Ethanolic and Aqueous Extracts of Myrtle (Myrtus communis) on Leishmania major

Background: The extract of myrtle plant contains polyphenolic compounds that show antibacterial, antiviral, and anti-parasitic properties. We aimed to investigate the therapeutic effect of aqueous and ethanolic myrtle extract against leishmaniasis caused by L. major in vivo and in vitro conditions. Methods: This study was carried out in Tarbiat Modares University, Tehran, Iran in 2018. Aqueous and ethanolic extract of myrtle plant at 6.25 to 400 mg/ml concentrations were tested on Leishmania major promastigotes, non-infected macrophages, and macrophages infected with amastigotes in vitro using counting, MTT and flow cytometry techniques. Then, BALB/c mice were treated with ethanolic, aqueous and a mixture of both extracts of myrtle plant. The treatment was carried out for four weeks. Then, the effectiveness of the herbal medicine was assessed by measuring wounds diameters, mice weights and their mortality rate on weekly basis. Results: The IC50 values of aqueous and ethanolic extracts for promastigotes were 7.86 and 11.66 μg/mL respectively. The IC50 values of the aqueous and ethanolic extracts for amastigotes were 12.5 and 47.2 μg/mL respectively. Flow cytometry indicates 62.88% and 60.16% apoptosis induced by ethanolic and aqueous extract of myrtle plant respectively. The lowest parasitic load was seen in the group treated with ethanolic extract. Conclusion: The lesion sizes for treated groups with extracts were similar to those treated with glucantime. Oral administration instead of injection is another advantage of myrtle plant over glucantime, which makes the herb easy and more practical.

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Evaluating the Effect of Cinnarizine on Promastigotes and Amastigotes forms of Leishmania major

Infectious Disorders - Drug Targets ◽

10.2174/1871526518666181113114820 ◽

2020 ◽

Vol 20 (4) ◽

pp. 550-555 ◽

Cited By ~ 1

Author(s):

Lima Asgharpour Sarouey ◽

Parvaneh Rahimi-Moghaddam ◽

Fatemeh Tabatabaie ◽

Khadijeh Khanaliha

Keyword(s):

Flow Cytometry ◽

Cutaneous Leishmaniasis ◽

Leishmania Major ◽

Inhibitory Effect ◽

Control Group ◽

Secondary Infections ◽

Ic50 Values ◽

J774 Cells ◽

Mtt Assays

: As an important global disease, cutaneous leishmaniasis is associated with complications such as secondary infections and atrophic scars. The first line treatment with antimonials is expensive and reported to have serious side effects and enhance resistance development. The main objective of this study was to evaluate the effect of Cinnarizine on standard strains of Leishmania major because of paucity of information on this subject. Methods: In this experimental study, four concentrations of the drug (5, 10, 15 and 20 μg/ml) were added to Leishmania major cultures at 24, 48 and 72 hours intervals. MTT assays were performed to determine parasite viability and drug toxicity. Leishmania major promastigotes were augmented to the in vitro cultured macrophages (J774 cells) and then incubated for 72 hours. Half maximal inhibitory concentration (IC50) was ascertained by counting parasites. The inhibitory effect of the drug was compared with that of Glucantime. Flow-cytometry was performed to investigate apoptosis. Each test was repeated thrice. Results: The IC50 values of Cinnarizine after 72 hours were calculated to be 34.76 μg/ml and 23.73 μg/ml for promastigotes and amastigotes, respectively. The results of MTT assays showed 48 % promastigote viability after 72 hour-exposure to Cinnarizine at 20 μg/ml concentration. Programmed cell death in promastigote- and amastigote-infected macrophages was quantified to be 13.66 % and 98.7 %, respectively. Flow- cytometry analysis indicated that Cinnarizine induced early and late apoptosis in parasites. All treatments produced results which differed significantly from control group (P<0.05). Conclusion: Cinnarizine showed low toxicity with anti-leishmanial and apoptosis effects on both promastigote and intracellular amastigote forms. Therefore, we may suggest further assessment on animal models of this drug as candidates for cutaneous leishmaniasis therapy.

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Characterization and selective inhibition of myristoyl-CoA:protein N-myristoyltransferase from Trypanosoma brucei and Leishmania major

Biochemical Journal ◽

10.1042/bj20051886 ◽

2006 ◽

Vol 396 (2) ◽

pp. 277-285 ◽

Cited By ~ 46

Author(s):

Chrysoula Panethymitaki ◽

Paul W. Bowyer ◽

Helen P. Price ◽

Robin J. Leatherbarrow ◽

Katherine A. Brown ◽

...

Keyword(s):

Trypanosoma Brucei ◽

Leishmania Major ◽

Homo Sapiens ◽

Selective Inhibition ◽

Fungal Species ◽

Chemotherapeutic Agents ◽

Human Pathogens ◽

Ic50 Values

The eukaryotic enzyme NMT (myristoyl-CoA:protein N-myristoyltransferase) has been characterized in a range of species from Saccharomyces cerevisiae to Homo sapiens. NMT is essential for viability in a number of human pathogens, including the fungi Candida albicans and Cryptococcus neoformans, and the parasitic protozoa Leishmania major and Trypanosoma brucei. We have purified the Leishmania and T. brucei NMTs as active recombinant proteins and carried out kinetic analyses with their essential fatty acid donor, myristoyl-CoA and specific peptide substrates. A number of inhibitory compounds that target NMT in fungal species have been tested against the parasite enzymes in vitro and against live parasites in vivo. Two of these compounds inhibit TbNMT with IC50 values of <1 μM and are also active against mammalian parasite stages, with ED50 (the effective dose that allows 50% cell growth) values of 16–66 μM and low toxicity to murine macrophages. These results suggest that targeting NMT could be a valid approach for the development of chemotherapeutic agents against infectious diseases including African sleeping sickness and Nagana.

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Antioxidant and Hypoglycemic Properties of Seeds of Cucurbitaceae Family from Western Nepal

Journal of Health and Allied Sciences ◽

10.37107/jhas.118 ◽

2019 ◽

Vol 9 (2) ◽

pp. 1-5

Author(s):

Atisammodavardhana Kaundinnyayana ◽

Anita G.C ◽

Asmita Banstola ◽

Kishwor Poudel ◽

Nirmal Acharya ◽

...

Keyword(s):

Antioxidant Activity ◽

Radical Scavenging ◽

Ethanolic Extract ◽

Positive Control ◽

Lagenaria Siceraria ◽

Benincasa Hispida ◽

Ic50 Values ◽

In Vitro Antioxidant Activity

Introduction: Diabetes mellitus is an endocrine and metabolic disorder characterized by the defect in insulin secretion or insulin action or both. The association between diabetes mellitus and oxidative stress is well established. Cucurbitaceae is the largest family containing 120 genera mostly grown for its sweet and juicy fruit in warm climates all over the world. In the present work ethanolic extracts of the seeds of Trichosanthes cucumerina Linn (Chichinno), Lagenaria siceraria (Lauka), Cucurbita pepo (Pharsi), Luffa aegiptiaca (Ghiraula), and Benincasa hispida (Kubindo) collected from Western Nepal has been evaluated for their in vitro antioxidant activity and in vivo hypoglycemic effects. Methods: In vitro antioxidant activity was assessed by using DPPH free radical scavenging activities and their IC50 values were calculated. In vivo hypoglycemic effect was examined on normoglycemic rats. The clinical significance of ethanolic extract at the doses of 250 and 500 mg/kg body weight was investigated in 0, 30, 60, 120 and 180 minutes of oral administration. Metformin treated group was used as the positive control. Results: For DPPH radical scavenging action, the IC50 values of Trichosanthes cucumerina, Luffa aegiptiaca, Benincasa hispida, Cucurbita pepo, Lagenaria siceraria were found to be 60.72, 127.73, 49.63, 98.16 and 52.46 µg/ml respectively which were compared to the IC50 value of ascorbic acid (Positive control) which was found to be 38.11 µg/ml. The extracts of Benincasa hispida, and were having higher antioxidant activity and tested for in vivo hypoglycemic activity.In vivo administration of two doses of ethanolic extract of Lagenaria siceraria reduced the level of blood glucose while the best result was obtained at 250 mg/kg. Conclusions: Present study revealed promising antioxidant and hypoglycemic activity of ethanolic extract of Lagenaria siceraria. The further exploration of Lagenaria siceraria for its effective use in the traditional medicinal system is essential.

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Inhibition Of Nuclear Transport Modulator Xpo1/CRM1 For The Treatment Of Acute Myeloid Leukemia (AML)

Blood ◽

10.1182/blood.v122.21.237.237 ◽

2013 ◽

Vol 122 (21) ◽

pp. 237-237 ◽

Cited By ~ 2

Author(s):

Michael P. Rettig ◽

Matthew Holt ◽

Julie Prior ◽

Sharon Shacham ◽

Michael Kauffman ◽

...

Keyword(s):

Flow Cytometry ◽

Board Of Directors ◽

Cell Lines ◽

Nuclear Export ◽

Employment Equity ◽

Advisory Committees ◽

Ic50 Values ◽

Equity Ownership

Abstract Background Exportin 1 (XPO1) also called CRM1, is a widely expressed nuclear export protein, transporting a variety of molecules including tumor suppressor proteins and cell cycle regulators. Targeted inhibition of XPO1 is a new strategy to restore multiple cell death pathways in various malignant diseases. SINEs are novel, orally available, small molecule Selective Inhibitors of Nuclear Export (SINE) that specifically bind to XPO1 and inhibit its function. Methods We used WST-1 cell proliferation assays, flow cytometry, and bioluminescence imaging to evaluate the efficacy of multiple SINEs to induce apoptosis alone and in combination with cytarabine (AraC) or doxorubicin in vitro in chemotherapy sensitive and resistant murine acute promyelocytic leukemia (APL) cells. This murine model of APL was previously generated by knocking in the human PML-RARa cDNA into the 5’ regulatory sequence of the cathepsin G locus (Westervelt et al. Blood, 2003). The abnormal co-expression of the myeloid surface antigen Gr1 and the early hematopoietic markers CD34 and CD117 identify leukemic blasts. These Gr1+CD34+CD117+ APL cells partially retain the ability to terminally differentiate toward mature granulocytes (mimicking more traditional AML models) and can be adoptively transferred to secondary recipients, which develop a rapidly fatal leukemia within 3 weeks after tumor inoculation. To assess the safety and efficacy of SINEs in vivo, we injected cryopreserved APL cells intravenously via the tail vein into unconditioned genetically compatible C57BL/6 recipients and treated leukemic and non-leukemic mice (n=15/cohort) with 15 mg/kg of the oral clinical staged SINE KPT-330 (currently in Phase 1 studies in patients with solid tumors and hematological malignancies) alone or in combination with 200 mg/kg cytarabine every other day for a total of 2 weeks. Peripheral blood was obtained weekly from mice for complete blood counts and flow cytometry to screen for development of APL. Results The first generation SINE, KPT214, inhibited the proliferation of murine APL cell lines in a dose and time dependent manner with IC50 values ranging from of 95 nM to 750 nM. IC50 values decreased 2.4-fold (KPT-185) and 3.5-fold (KPT-249) with subsequent generations of the SINEs. Consistent with the WST-1 results, Annexin V/7-aminoactinomycin D flow cytometry showed a significant increase of APL apoptosis within 6 hours of KPT-249 application. Minimal toxicity against normal murine lymphocytes was observed with SINEs even up to doses of 500 nM. Additional WST-1 assays using AraC-resistant and doxorubicin-resistant APL cell lines demonstrated cell death of both chemotherapy-resistant cell lines at levels comparable to the parental chemosensitive APL cell lines. Combination therapy with low dose KPT-330 and AraC showed additive effects on inhibition of cell proliferation in vitro. This additive effect of KPT-330 and chemotherapy on APL killing was maintained in vivo. As shown in Figure 1, treatment with AraC or KPT-330 alone significantly prolonged the survival of leukemic mice from a median survival of 24 days (APL + vehicle) to 33 days or 39 days, respectively (P < 0.0001). Encouragingly, combination therapy with AraC + KPT-330 further prolonged survival compared to monotherapy (P < 0.0001), with some mice being cured of the disease. Similar in vivo studies with the AraC-resistant and doxorubicin-resistant APL cells are just being initiated. Conclusions Our data suggests that the addition of a CRM1 inhibitor to a chemotherapy regimen offers a promising avenue for treatment of AML. Disclosures: Shacham: Karyopharm Therapeutics Inc.: Employment, Equity Ownership, Membership on an entity’s Board of Directors or advisory committees, Patents & Royalties. Kauffman:Karyopharm Therapeutics Inc.: Employment, Equity Ownership, Membership on an entity’s Board of Directors or advisory committees, Patents & Royalties. McCauley:Karyopharm Therapeutics, Inc: Employment, Equity Ownership.

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Invitro Antimicrobial Activities of Mitracarpus scaber Against Some Common Bacteria of Aquatic Origin

Media Kedokteran Hewan ◽

10.20473/mkh.v32i3.2021.119-130 ◽

2021 ◽

Vol 32 (3) ◽

pp. 119

Author(s):

Abdullateef Abiodun Ajadi ◽

Benjamin Emikpe ◽

Ahmed Akeem

Keyword(s):

Antimicrobial Properties ◽

Ethanolic Extract ◽

Antibacterial Activities ◽

Antimicrobial Activities ◽

Phytochemical Analysis ◽

Ethanolic Extracts ◽

Zones Of Inhibition ◽

Mitracarpus Scaber

Some plants have been reported to be of medicinal values and reserve some antimicrobial properties. One of such plants is Mitracarpus scaber and its effect on bacterial growth is evaluated. The study aimed at evaluating the phytochemical analyses and antimicrobial potentials of Mitracarpus scaber against aquatic bacteria including Aeromonas, Bacillus, Vibrio and Staphylococcus Spp. Leaves of Mitracarpus scaber were collected, washed and air dried and phytochemical analysis and antimicrobial investigation of ethanolic and aqueous extracts of the leaves were carried out against a panel of bacteria isolated from diseased catfish from various farms. The phytoconstituents detected include saponins, tannins, flavonoids, tarpenoids, steroids, anthraquinones and alkaloids in both aqueous and ethanolic extracts. Both aqueous and ethanolic extracts of M. scaber showed varying degree of antibacterial activities but ethanolic extract showed a higher activity against the pathogens tested. The ethanolic extract had zones of inhibition similar to that of standard antibiotics (enrofloxacin) across all tested microbes. The lowest minimum inhibitory concentration of ethanolic extract of M. scaber was against Bacillus sp with 10mg/ml while the highest was 85mg/ml against Staphylococcus species. The results of the assays showed promising evidences that M. scaber is a potential antibacterial agent against aquatic microbes. However, further studies are recommended to fractionate its constituents and determine the in vitro and in vivo anti-microbial activities and the exact mechanism of action of the constituents.

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In vivo and In vitro effects of ethanolic extract of Trigonella foenum-graecum L. seeds on proliferation, angiogenesis and tube formation of endothelial cells

Research in Pharmaceutical Sciences ◽

10.4103/1735-5362.235161 ◽

2018 ◽

Vol 13 (4) ◽

pp. 343 ◽

Cited By ~ 4

Author(s):

Mohammad Soukhtanloo ◽

Mozhdeh Iranmanesh ◽

Reza Mohebbati ◽

Fatemeh Forouzanfar ◽

MostafaKarimi Roshan ◽

...

Keyword(s):

Endothelial Cells ◽

Ethanolic Extract ◽

Tube Formation ◽

Trigonella Foenum Graecum ◽

In Vitro Effects ◽

Trigonella Foenum Graecum L

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MicroRNAs Expression Induces Apoptosis of Macrophages in Response to Leishmania major (MRHO/IR/75/ER): An In-Vitro and In-Vivo Study

Iranian Journal of Parasitology ◽

10.18502/ijpa.v15i4.4851 ◽

2020 ◽

Author(s):

Mostafa GHOLAMREZAEI ◽

Soheila ROUHANI ◽

Mehdi MOHEBALI ◽

Samira MOHAMMADI-YEGANEH ◽

Mostafa HAJI MOLLA HOSEINI ◽

...

Keyword(s):

Flow Cytometry ◽

Leishmania Major ◽

Parasite Burden ◽

Parasite Load ◽

Lesion Size ◽

In Vivo Studies ◽

Raw 264.7 Cells ◽

Control Group

Background: We aimed to investigate the effect of miR-15a mimic and inhibitor of miR-155 expression on apoptosis induction in macrophages infected with Iranian strain of Leishmania major in-vitro and in-vivo. Methods: RAW 264.7 cells were infected with L. major promastigotes (MRHO/IR/75/ER), and then were treated with miRNAs. For in-vivo experiment, BALB/c mice were inoculated with L. major promastigotes, and then they were treated with miRNAs. For evaluation of miRNA therapeutic effect, in-vitro and in-vivo studies were performed using quantitative Real-time PCR, Flow cytometry, lesion size measurement, and Limiting Dilution Assay (LDA). This study was performed in Shahid Beheshti University of Medical Sciences in 2019. Results: In-vitro results of flow cytometry showed that using miR-15a mimic, miR-155 inhibitor or both of them increased apoptosis of macrophages. In in-vivo, size of lesion increased during experiment in control groups (P<0.05) while application of both miR-155 inhibitor and miR-15a mimic inhibited the increase in the size of lesions within 6 wk of experiment (P=0.85). LDA results showed that microRNA therapy could significantly decrease parasite load in mimic or inhibitor receiving groups compared to the control group (P<0.05). Conclusion: miR-155 inhibitor and miR-15a mimic in L. major infected macrophages can induce apoptosis and reduce parasite burden. Therefore, miRNA-based therapy can be proposed as new treatment for cutaneous leishmaniasis.

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Silybum marianum ethanolic extract: in vitro effects on protoscolices of Echinococcus granulosus G1 strain with emphasis on other Iranian medicinal plants

Tropical Medicine and Health ◽

10.1186/s41182-021-00363-7 ◽

2021 ◽

Vol 49 (1) ◽

Author(s):

Ali Taghipour ◽

Fatemeh Ghaffarifar ◽

John Horton ◽

Abdolhossein Dalimi ◽

Zohreh Sharifi

Keyword(s):

Medicinal Plants ◽

Echinococcus Granulosus ◽

High Performance ◽

Hplc Analysis ◽

Ethanolic Extract ◽

Silybum Marianum ◽

Wide Range ◽

In Vitro Effects

Abstract Background Cystic echinococcosis (CE), is a parasitic zoonosis caused by Echinococcus granulosus (E. granulosus) larvae in liver and lungs of both humans and animals. Surgical intervention is the mainstay for CE treatment, using scolicidal agents that inactivate live protoscolices. This study evaluated the scolicidal effects of Silybum marianum ethanolic extract and its combination with albendazole in vitro for the first time. Moreover, in a literature review, we investigated the effects of a wide range of Iranian medicinal plants on protoscolices of E. granulosus. Methods S. marianum ethanolic extract was prepared and high-performance liquid chromatography (HPLC) analysis was used to establish the proportions of its component compounds in the extract. Cytotoxicity was evaluated in mouse macrophage cells (J774A.1 cell line) using MTT method. Next, the scolicidal activity of the extract alone and combined with albendazole was tested as triplicate at various concentrations incubated for 5, 10, 20, 30, and 60 min. Finally, protoscolex viability was determined using 0.1% eosin as a vital stain. PCR–RFLP and DNA sequencing techniques were used to characterize the genotype of E. granulosus. Results HPLC analysis showed that S. marianum ethanolic extract contained mostly silydianin (14.41%), isosilybin A (10.50%), and silychristin (10.46%). The greatest scolicidal effects were obtained with the combination of S. marianum with albendazole (79%), S. marianum ethanolic extract alone (77%) and albendazole (69%), at a concentration of 500 μg/ml for 60 min, respectively (P < 0.05). Molecular analysis showed that all the cysts used were G1 genotype. Conclusion The data suggest that S. marianum ethanolic extract is a potential scolicide in vitro; however, further investigations are required to determine its efficacy in vivo.

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