Calendula arvensis L. as an anti-cancer agent against breast cancer cell lines.

Calendula arvensis L. is used in traditional folk medicine for the treatment of several diseases. Leaves, stems, and flowers of C. arvensis were extracted using a Soxhlet extractor with different solvents (i.e., hexane, chloroform, ethyl acetate, and methanol). The ethyl acetate extract of C. arvensis flowers (CAF EtOAC) had cytotoxic activity against MCF-7 and MDA-MB-231 breast cancer cells, with IC50 values of 70 and 78 µg/mL, respectively. Microscopic examination revealed concentration-dependent cell shrinkage, cell detachment, nuclear fragmentation, and chromatin condensation. The CAF EtOAC inhibited the migration of cultured cells in a scratch wounding assay, indicating a possible defense against metastasis. The same extract also caused apoptosis by downregulating Bcl-2 and upregulating Bax and caspase 3/7 activity. Phytochemical analyses revealed the presence of phenols and flavonoids, and gas chromatography-mass spectroscopy (GC-MS) revealed a high content of linolenic acid in the extract. Based on our data, the CAF EtOAC may provide active ingredients for the development of novel chemotherapeutics for breast cancer therapy.

Bitter gourd (Momordica charantia) possess developmental toxicity as revealed by screening the seeds and fruit extracts in zebrafish embryos.

BACKGROUND: Bitter gourd (Momordica charantia) has attracted the focus of researchers owing to its excellent anti-diabetic action. The beneficial effect of Momordica charantia on heart has been reported by in vitro and in vivo studies. However the developmental toxicity or potential risk of M. charantia on fetus heart development is largely unknown. Hence this study was designed to find out the developmental toxicity of M. charantia using zebrafish (Danio rerio) embryos. METHODS: The crude extracts were prepared from fruit and seeds of M. charantia. The Zebrafish embryos were exposed to serial dilution of each of the crude extract. The biologically active fractions were fractionated by C18 column using high pressure liquid chromatography. Fourier-transform infrared spectroscopy and gas chromatography coupled with mass spectrophotometry was done to identify chemical constituents in fruit and seed extract of M. charantia. RESULTS: The seed extract of M. charantia was lethal with LD50 values of 50 µg/ml to zebrafish embryos and multiple anomalies were observed in zebrafish embryos at sub-lethal concentration. However, the fruit extract was much safe and exposing the zebrafish embryos even to 200 µg/ml did not result any lethality. The fruit extract induced severe cardiac hypertrophy in treated embryos. The time window treatment showed that M. charantia perturbed the cardiac myoblast specification process in treated zebrafish embryos. The Fourier-transform infrared spectroscopy analyses revealed diverse chemical group in the active fruit fraction and five new type of compounds were identified in the crude seeds extract of M. charantia by gas chromatography and mass spectrophotometry. CONCLUSION: The teratogenicity of seeds extract and cardiac toxicity by the fruit extract of M. charantia warned that the supplementation made from the fruit and seeds of M. charantia should be used with much care in pregnant diabetic patients to avoid possible damage to developing fetus.

Apoptotic Induction and Anti-Migratory Effects of Rhazya Stricta Fruit Extracts on a Human Breast Cancer Cell Line.

Rhazya stricta is a medicinal plant that is widely used in Saudi folklore medicine for treatment of various diseases. R. stricta fruit powder was sequentially extracted with n-hexane, chloroform, ethyl acetate, and methanol using a Soxhlet extractor. The cytotoxic effects of these fractions on human breast cancer cells (MDA-MB-231 and MCF-7) and non-tumorigenic control cells (MCF-10A) were evaluated via cell viability measurements, microscopy, gene expression, and migration assays. Moreover, the effect of the most promising extract on 7,12-dimethyl-benz[a]anthracene (DMBA)-induced breast cancer was investigated in rats. The promising extract was also subjected to gas chromatography-mass spectrometry. Fruit extracts of R. stricta were significantly cytotoxic toward all tested cell lines, as demonstrated by MTT and LDH assays. Treatment of MDA-MB-231 cells with fruit ethyl acetate fraction (RSF EtOAc) increased expression 11of P53, Bax and activation of caspase 3/7. A cell migration scratch assay demonstrated that extracts at non-cytotoxic concentrations exerted a potent anti-migration activity against the highly invasive MDA-MB-231 cell line. Moreover, RT-PCR results showed that RSF EtOAc significantly downregulated MMP-2 and MMP-9 expression, which play an important role in breast cancer metastasis. Histological studies of breast tissue in experimental animals showed a slight improvement in tissue treated with fruit ethyl acetate extract. GC-MS chromatogram showed thirteen peaks with major constituents were camphor, trichosenic acid and guanidine. Our current study demonstrates that fruit extracts of R. stricta are cytotoxic toward breast cancer cell lines through apoptotic mechanisms.

Analysis of Chemical Composition and Assessment of Cytotoxic, Antimicrobial, and Antioxidant Activities of the Essential Oil of Meriandra dianthera Growing in Saudi Arabia.

The essential oil of Meriandra dianthera (Konig ex Roxb.) Benth. (Synonym: Meriandra bengalensis, Lamiaceae) collected from Saudi Arabia was studied utilizing GC and GC/MS. Forty four constituents were identified, representing 96.8% of the total oil. The M. dianthera essential oil (MDEO) was characterized by a high content of oxygenated monoterpenes (76.2%). Camphor (54.3%) was the major compound in MDEO followed by 1,8-cineole (12.2%) and camphene (10.4%). Moreover, MDEO was assessed for its cytotoxic, antimicrobial, and antioxidant activities. MDEO demonstrated an interesting cytotoxic activity against all cancer cell lines with IC50 values of 83.6 to 91.2 µg/mL, especially against MCF-7 cancer cells. Using labeling with annexin VFITC and/or propidium iodide (PI) dyes and flow cytometer analysis, the apoptosis induction was quantitatively confirmed for MCF-7 cells. The MDEO exhibited a considerable antimicrobial activity against all bacterial and fungal strains with minimum inhibitory concentration (MIC)-values of 0.07 to 1.25 mg/mL. The most sensitive microbial strain was Staphylococcus aureus (MIC: 0.07 mg/mL). Minimum bactericidal concentration (MBC) or minimum fungicidal concentration (MFC) values were determined one time higher than that of MIC's. Additionally, the MDEO revealed a strong activity for reducing ß-carotene bleaching with a total antioxidant value of 72.6% and significant DPPH free radical scavenging activity (78.4%) at the concentration 1000 µg/mL.

Anti-proliferative and anti-inflammatory activities of entophytic Penicillium crustosum from Phoenix dactylifer.

Natural sources have been and will remain an inspiration source for modern chemistry. The current study investigates the antiproliferative and anti-inflammatory action of the ethyl acetate fraction of Penicillium crustosum from Phoenix dactylifera. This paper reports the isolation of P. crustosum from leaves of P. dactylifera and the antiproliferative activities of ethyl acetate fraction on cancer cells. To reach this goal, the anti-proliferation and cytotoxicity effects were evaluated by MTT and LDH assay respectively. The quantitative real time PCR technique was used to investigate IL-6 and IL-8 gene expression. Our results revealed higher anti-proliferative activity against HepG2 (82µg/ml) than MCF7 (126µg/ml) and inhibited the migration of the cell lines. The ethyl acetate fraction significantly altered LDH levels and reduced IL-6 transcript expression on MCF7 cell line but not in HepG2 cell line which could be specific anti-inflammatory drug in breast cancer cell line. These results suggest that Phoenix dactylifera extract has a potent anti-proliferative and anti-inflammatory action. Further investigation to isolate the active compounds and mode of action is required.

In vitro induction of human embryonal carcinoma differentiation by a crude extract of Rhazya stricta.

BACKGROUND: Rhazya stricta Decne. is a medicinal plant that is widespread in Saudi Arabia and desert areas of the Arabian Peninsula. Its extract contains alkaloids, tannins, and flavonoids that are involved in different biological activities. The study aim was to evaluate the effects of Rhazya stricta plant extracts on the proliferation and differentiation of NTERA-2 (NT2) pluripotent embryonal carcinoma cells. METHODS: Soxhlet extraction was carried out using different solvents to extract stems, leaves and fruit parts of this plant. Cytotoxicity was evaluated by an MTS cell viability assay. The ability of the plant extract to induce cell differentiation was examined phenotypically using an inverted light microscope. The expression of pluripotency markers was investigated by reverse transcriptase polymerase chain reaction (RT-PCR) and immunocytochemistry. Phytochemical screening of chloroform stem extracts was carried out and a chromatographic fingerprint was generated using gas chromatography - mass spectrometry (GC-MS). RESULTS: Chloroform stem extract induced differentiation of NT2 cells at 5 µg/ml, and the differentiated cells exhibited neurite formation. Following induction of differentiation, there was significant down-regulation of the pluripotency marker genes Oct4 and Sox2. In addition, the surface antigen pluripotency marker, TRA-1-60, was strongly down-regulated. Phytochemical analysis of the extract showed the presence of alkaloids and saponins. The chromatogram revealed the presence of fifteen compounds with different retention times. CONCLUSION: Our results demonstrate for the first time that chloroform stem extract of R. stricta can induce neuronal differentiation of stem cells at an early stage and may contain potential therapeutic agent that can be used in neurodegenerative diseases.

Synthesis, Characterization, and Anti-Cancer Activity of Some New N'-(2-Oxoindolin-3-ylidene)-2-propylpentane hydrazide-hydrazones Derivatives.

Eight novel N'-(2-oxoindolin-3-ylidene)-2-propylpentane hydrazide-hydrazone derivatives 4a-h were synthesized and fully characterized by IR, NMR ((1)H-NMR and (13)C-NMR), elemental analysis, and X-ray crystallography. The cyto-toxicity and in vitro anti-cancer evaluation of the prepared compounds have been assessed against two different human tumour cell lines including human liver (HepG2) and leukaemia (Jurkat), as well as in normal cell lines derived from human embryonic kidney (HEK293) using MTT assay. The compounds 3e, 3f, 4a, 4c, and 4e revealed promising anti-cancer activities in tested human tumour cells lines (IC50 values between 3 and 7 µM) as compared to the known anti-cancer drug 5-Fluorouracil (IC50 32-50 µM). Among the tested compounds, 4a showed specificity against leukaemia (Jurkat) cells, with an IC50 value of 3.14 µM, but this compound was inactive in liver cancer and normal cell lines.

In vitro antiproliferative activity of partially purified Withania somnifera fruit extract on different cancer cell lines.

PURPOSE: Cancer is a major health problem worldwide. There is a continuous need to search for safer and more effective alternatives to overcome the side effects and resistance of the chemotherapeutic agents. Therefore, in this study we investigated the antiproliferative activity and the apoptotic potential of Withania somnifera (W. somnifera). METHODS: W. somnifera was extracted with methanol and then solvent partitioned by sequential extractions with hexane, dichloromethane and ethyl acetate. Each extract was assayed for antiproliferative activity against different cancer cell lines using MTT assay. The nuclear morphology of HepG2 cells was investigated by DNA-binding fluorescent dye (Hoechst 33342 stain). The percentage of viability, death and apoptosis were evaluated by the Tali(TM) Image-based cytometer using annex-in-V/PI (propidium iodide). A chromatographic fingerprint was constructed using high performance liquid chromatography (HPLC). RESULTS: The most potent anticancer activity of the crude extract was against HepG2 cell line (LC50=164.7µg/ml). Dichloromethane fraction showed remarkable changes in the chromatin structure i.e., fragmentation, uniform condensation. Of the HepG2 cells 43.6% were apoptotic when treated with dichloromethane fraction for 24 hrs at 95µg/ml concentration. HPLC showed the presence of a major peak at 11.85 min. CONCLUSION: W. somnifera may have the potential to serve as a template for future anticancer drug development. However, further investigation is required to identify the active compound/s.

Synthesis and biological activity of Schiff base series of valproyl, N-valproyl glycinyl, and N-valproyl-4-aminobenzoyl hydrazide derivatives.

Series of Schiff bases of valproic acid hydrazide, N-valproylglycine hydrazide and N-valproyl-4-aminobenzoyl hydrazide derivatives were synthesized and characterized by IR, NMR ((1)H- and (13)C-NMR) and elemental analysis. The prepared compounds were evaluated in transgenic zebrafish embryos (Tg: flil-1: enhanced green fluorescent protein (EGFP)) for antiangiogenic activity and in HepG2 liver carcinoma cell line for anti cancer activity. The Schiff bases of N-valproylglycine hydrazide derivatives were most potent in term of suppressing angiogenic blood vessels formation and anticancer activity at very low concentration, followed by the Schiff bases of valproic acid hydrazide derivatives which exhibited moderate activity, while the Schiff bases of N-valproyl-4-aminobenzoyl hydrazide derivatives, ethyl 4-(2-propylpentanamido)benzoate (VABE) and N-(4-(hydrazinecarbonyl)phenyl)-2-propylpentanamide (VABH) in contrast exhibited pro-angiogenic activity and weaker anticancer activity which mean that these derivatives targeted a common signaling pathway in term of affecting the blood vessels formation.

Evaluation of the Anticancer Potential of Morus nigra and Ocimum basilicum Mixture against Different Cancer Cell Lines: An In Vitro Evaluation.

Morus nigra (M) and Ocimum basilicum (O) mixture (MO2) extract was extracted using hexane (MO2H), chloroform (MO2C), ethyl acetate (MO2E), and methanol (MO2M) in a Soxhlet apparatus. The cytotoxicity was evaluated using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. The IC50 values of the MO2C-treated cancer cells were 11.31 µg/mL (MDA-MB-231), 15.45 µg/mL (MCF-7), 18.9 µg/mL (HepG2), 26.33 µg/mL (Huh-7), 30.17 µg/mL (LoVo), and 36.76 µg/mL (HCT116). MO2C-treated cells showed cellular and nuclear morphological alterations like chromatin condensation and formation of apoptotic bodies as observed using light and fluorescent microscopy. The antioxidant and anti-inflammatory properties were investigated in vitro using 2,2'-diphenyl-1-picrylhydrazyl (DPPH) and egg albumin denaturation assays. It was evident that the MO2M extract exhibited the highest antioxidant activity (18.13%), followed by the MO2E extract (12.25%), MO2C extract (9.380%), and MO2H extract (6.31%). The highest inhibition percentage of albumin denaturation was observed in MO2H (28.54%), followed by MO2M (4.32%) at 0.2 and 0.1 mg/mL concentrations, respectively. The compounds identified using gas chromatography-mass spectrometry (GC-MS) analysis for MO2C extract were α-trans-bergamotene, germacrene D, selin-4,7(11)-diene, 2 tridecen-1-ol, and 2-decen-1-ol. The present study reveals that MO2C has promising anticancer activity and may serve as a potent polyherbal extract in cancer treatment.

The molecular interplay of known phytochemicals as Culex pipiens and Rift Valley fever virus inhibitors through molecular docking.

Infectious diseases transmitted by vectors have claimed millions of lives. The mosquito Culex pipiens is a main vector species of Rift Valley Fever virus (RVFV) transmission. RVFV is an arbovirus that infects both people and animals. No effective vaccine or drugs are available for RVFV. Therefore, it is vital to find effective therapies for this viral infection. Because of their critical roles in transmission and infection, acetylcholinesterase 1 (AChE1) of Cx. Pipiens and RVFV glycoproteins, and nucleocapsid proteins are appealing protein targets. To understand intermolecular interactions, computational screening was carried out using molecular docking. More than 50 compounds were tested against different target proteins in the current study. Anabsinthin (-11.1 kcal/mol), zapoterin (-9.4 kcal/mol), porrigenin A (-9.4 kcal/mol), and 3-Acetyl-11-keto-beta-boswellic acid (AKBA) (-9.4 kcal/mol) were the top hit compounds for Cx. Pipiens. Similarly, the top hit compounds for RVFV were zapoterin, porrigenin A, anabsinthin, and yamogenin. The toxicity of Rofficerone is predicted as fatal (Class II), whereas Yamogenin is safe (Class VI). Further investigations are needed to validate the selected promising candidates against Cx. pipiens and RVFV infection using in-vitro and in-vivo methods.

Corrigendum to "Biogenic synthesis of silver nanoparticles: Antibacterial and cytotoxic potential' [Saudi J. Biol. Sci. 27(5) (2020) 1340-1351].

[This corrects the article DOI: 10.1016/j.sjbs.2019.12.014.].

Larvicidal activity and Histopathological changes of Cinnamomum burmannii, Syzygium aromaticum extracts and their combination on Culex pipiens.

In order to develop an eco-friendly botanical larvicide alternative to the synthetic larvicides, extracts were prepared from the Cinnamomum burmannii (C.B.) and Syzygium aromaticum (S.A.) with hexane using a sonicator. The extracts were evaluated for larvicidal activity individually and in combination against the Culex pipiens larvae. The LC50 value of C.B. and the S.A. hexane extracts tested individually were 184.2 and 363.7 µg/mL against Cx. pipiens respectively. All the combinations of the extract of C.B. and S.A. showed synergistic factors higher than one. Among the different ratios of extracts, the SA25%: CB75% extract was found to be more toxic than the other combinations (LC50:125.7 µg/mL). Midgut cells treated with S.A. 25%: C.B. 75% extract showed severe morphological alterations such as degradation of microvilli; degeneration of epithelial cells, and peritrophic membrane; loss of nuclei, irregular and damage of microvilli. The extract has a promising larvicidal potential against Cx. pipiens, However, the extract was toxic against HUVEC cells, as evident from MTT and cell morphology. Further investigation is required to assess the toxicity of the extract on aquatic animals.

Insecticidal effects of a novel polyherbal formulation (HF7) against Culex pipiens L. (Diptera: Culicidae).

Plant secondary metabolites represent the most efficient and convenient method to control and overcome environmental pollution and insecticidal resistance. This study explored the mosquitocidal activity of the combined extract of seven plants, (HF7) extracted using a Soxhlet extractor against Culex pipiens under laboratory conditions. Exposure of the 3rd instars of Cx. pipiens to HF7 hexane extract resulted in LC50:114.5 µg/mL and LC90:117.0 µg/mL values after 24 h. The ovicidal activities of hexane extract against Cx. pipiens eggs were 21.6%, 48.3%, and 71.6% at 187.5, 93.7, and 46.88 µg/mL, respectively. HF7-treated larvae showed the formation of irregular blebbing of epithelial cells toward the lumen and sloughing into the gut lumen. HF7 extract resulted in 100% adulticidal mortality at the concentration of 3.7 mg/test tube after 30 min of exposure. The IC50 of HF7 extract was 97.03 µg/ml against larvae, at which nuclear and morphological changes were observed. The spectroscopy spectrum of HF7 hexane extract disclosed the presence of 57 different secondary metabolites, among which the dominant compound was eugenol (32.3%). HF7 hexane extract could serve as a botanical insecticide for controlling Cx. pipiens and potentially other mosquito species.

Evaluating the efficacy of an innovative herbal formulation (HF6) on different human cancer cell lines.

Cancer is reported to be the leading cause of death and illness worldwide. This research aims to evaluate the phytochemicals, antioxidant, cytotoxic, and apoptotic activities of the polyherbal formulation HF6. HF6 was prepared by blending equal quantities of plants powder, namely, Curcuma longa, Salvia officinalis, Cinnamomum zeylanicum, Capsicum annuum, Zingiber officinale, and Syzygium aromaticum, and later extracted using hexane (HF6H), chloroform (HF6C), ethyl acetate (HF6E), and methanol (HF6M) in Soxhlet apparatus. Among the four different extracts, only the hexane extract (HF6H) was significantly effective. The HF6H extract showed antioxidant and anticancer potentials against different cancer cell lines, and moderate cytotoxicity against non-cancer cells, rendering it a promising remedy. In addition, it exerted tremendous cytotoxic effects on MCF-7, Huh-7, HCT116, MDA-MB-231, LoVo, and HepG2 cells with IC50 values of 2.02, 4.5, 6.9, 11.4, 23.5, and 34.7 µg/mL, respectively. The morphological hallmarks of apoptosis such as the rounding of cells, loss of contact with neighboring cells, formation of cell membrane blebbing, and microspike protrusion were detected using several different techniques. DAPI staining revealed apoptotic nuclear morphology such as condensation and DNA fragmentation. The morphological changes of MCF7 cells were also analyzed by AO/EB fluorescence staining. MCF7-stained green cells were viable cells, whereas the treated cells showed fragmented green nuclei representing early apoptosis. The phytochemical screening of HF6H showed positive results regarding the presence of alkaloids, polyphenols, flavonoids, and sterols. The GC-MS (gas chromatography-mass spectrometry) analysis of the HF6H extract indicated the presence of 12 compounds, mainly trans-caryophyllene (21.55%), cis-isoeugenol (18.42%), acetyleugenol (17.53%), alpha farnesene (10.0%), and zingiberene (8.55%). However, further investigation could be carried out to examine the toxicity of the extract on animal models.

Highly bioactive novel aryl-, benzyl-, and piperazine-selenoureas: synthesis, structural characterization and in vitro biological evaluation.

Selenoureas are widespread as useful elements for constructing important species and biologically active molecules. Finding an efficient and straightforward method to prepare this motif and biologically screen derivatives thereof is crucial. Herein, we demonstrate the effectiveness of using ethanol as a solvent in the preparation of various substituted aryl-, benzyl-, and piperazine-selenoureas from isoselenocyanates and amines. The synthetic method includes mild reaction conditions, large substrate scope, and good isolated yields. Biological evaluation of the prepared products on MDA-MB-231 and MCF-7 cancer cell lines revealed several remarkably active compounds (IC50 < 10 µΜ) with the best one exhibiting IC50 values of 1.8 µΜ and 1.2 µΜ observed against the challenging former triple-negative breast cancer cell line and the latter one, respectively. The chemical structures of all new compounds were fully characterized by multinuclear nuclear magnetic resonance (NMR) spectroscopy and high accuracy mass measurements.

Corrigendum to "Target and Non-target effects of Foeniculum vulgare and Matricaria chamomilla combined extract on Culex pipiens mosquitoes" [Saudi J. Biol. Sci. 28(10) (2021) 5773-5780].

[This corrects the article DOI: 10.1016/j.sjbs.2021.06.024.].

Cytotoxicity, in vivo toxicity, and chemical composition of the hexane extract of Plectranthus amboinicus (Lour.) Spreng.

Cancer is a universal health issue, and many anticancer therapeutic drugs have been isolated from natural products. This study analyzed the cytotoxic and apoptotic activity of Plectranthus amboinicus leaf hexane (PALH) extract in MDA-MB-231 (median inhibitory concentration [IC50] = 39.26 µg/mL) and MCF7 (IC50 = 89.05 µg/mL) breast cancer cell lines. Cells appeared rounded and shrunken, indicating morphological changes due to apoptosis induction. The primary constituent of PALH was phenol, 5-methyl-2-(1-methylethyl) (44%). PALH extract treatment increased the percentage of late apoptotic cells in the MDA-MB231 cell line (58% ± 1.5% at 200 µg/mL) compared to the control group, as evidenced by the activated caspase-3 and caspase-7 identified and captured by fluorescence microscopy. The relative migration rate in MDA-MB-231 cells treated with 10 µg/mL of PALH extract for 48 h was significantly lower compared to the control group. Analysis of acute (2000 mg/kg/BW) and subacute (250 and 500 mg/kg/BW) toxicity of PALH extract in mice showed no mortality or adverse effects in the kidney and liver histology compared to the control group. PALH extract can be considered nontoxic as it does not cause any adverse changes and so can be proposed as a potential breast anticancer agent.

Target and non-target effects of Foeniculum vulgare and Matricaria chamomilla combined extract on Culex pipiens mosquitoes.

The present study focused on extracting green larvicides from extracts of the combination of Foeniculum vulgare and Matricaria chamomilla using different solvents of increasing polarity in a Soxhlet extractor and evaluating their ovicidal, larvicidal, and cytotoxic activities. The most promising among all tested extracts was hexane extract. The ovicidal activity of the hexane PH2 extract resulted in a significant (p < 0.05) decrease in egg hatchability from 95.00 ± 6.16% to 15 ± 9.04% at doses ranging from 62.5 to 500 µg/mL. The larval mortality with the hexane extract ranged from 13.33 ± 3.3% to 93.33 ± 3.3% at doses ranging from 31.25 to 250 µg/mL, respectively. The LC50 and LC90 values of the larvicidal activity of the hexane extract were estimated to be 148.3 and 242.17 µg/mL, respectively, after 24 h of exposure. Similarly, the LC50 values after 48 and 72 h of exposure were 124.93 and 100.3 µg/mL, respectively, against the third instar of Cx. pipiens. PH2 treatment of larvae resulted in histopathological changes such as degenerated epithelial cells and destruction of microvilli on the epithelial cells. The PH2 extract achieved a dose-dependent decrease in the rate of cell survival. The IC50 value of PH2-treated HUVECs was 192.07 µg/mL after 24 h of incubation. The cells showed changes in cellular and nuclear morphology. In conclusion, the hexane extract of PH2 could be used in mosquito management programs.

Target and Nontarget Toxicity of Cassia fistula Fruit Extract Against Culex pipiens (Diptera: Culicidae), Lung Cells (BEAS-2B) and Zebrafish (Danio rerio) Embryos.

Mosquitoes transmit serious diseases, which threaten humans and severely affect livestock. The half-lethal concentration (LC50) was calculated by log probit analysis. The LC50 and LC90 values of larvicidal activity of Cassia fistula Linn. hexane-methanol soluble fraction (HMSF) after 24 h of exposure were 21.04 and 34.68 µg/ml, respectively. The LC50 values after 24 h of exposure were 84.09 µg/ml and 108.08 µg/ml for chloroform-methanol soluble fraction (CMSF) and ethyl acetate-methanol soluble fraction (EMSF) respectively. The percent hatchability of eggs exposed to the hexane extract was 90 ± 5.0, 68.33 ± 7.6, 46.6 ± 11.5, 10 ± 0.0, and 0 ± 0.0% at 10, 20, 40, 60, and 80 ppm, respectively. The pupicidal activity of the hexane extract at 40 µg/ml was 0.0%. The LC50 value of adulticidal activity of the hexane extract was 12.8 mg/test tube. The biosafety of the hexane extract was assessed in nontarget organisms, i.e., zebrafish (Danio rerio) embryos and normal lung cells (BEAS-2B). The hexane extract of C. fistula was well tolerated by zebrafish embryos, and no mortality or toxicity was found in the embryos exposed to the highest tested concentration of 300 µg/ml. Similarly, all the concentrations tested against the normal lung cells (BEAS-2B) showed more than 95% survival. The gas chromatography-mass spectroscopy analysis identified 12 compounds, and 2-methyl hexanoic acid and 2-methyl butanoic acid were the major compounds identified in the hexane extract. The larvicidal activity of C. fistula extracts will help in the development of natural substitutes for vector management of mosquito populations.