Tomato Leaf Curl New Delhi Virus Spain Strain Is Not Transmitted by Trialeurodes vaporariorum and Is Inefficiently Transmitted by Bemisia tabaci Mediterranean between Zucchini and the Wild Cucurbit Ecballium elaterium.

Tomato leaf curl New Delhi virus (ToLCNDV) is a bipartite begomovirus (genus Begomovirus, family Geminiviridae) persistently transmitted, as with all other begomoviruses, by whiteflies (Hemiptera: Aleyrodidae) of the Bemisia tabaci cryptic species complex. The virus, originally from the Indian subcontinent, was recently introduced in the Mediterranean basin, where it is currently a major concern for protected and open-field horticulture. The Mediterranean ToLCNDV isolates belong to a novel strain named "Spain strain" (ToLCNDV-ES), which infects zucchini and other cucurbit crops but is poorly adapted to tomato. Recently, it has been reported that another whitefly, Trialeurodes vaporariorum, is able to transmit an isolate of ToLCNDV from India which infects the chayote plant, a cucurbit. The present work aimed to clarify some aspects of whitefly transmission of ToLCNDV-ES. It was shown that T. vaporariorum is not able to transmit ToLCNDV-ES between zucchini plants. In addition, Ecballium elaterium may not act as a relevant reservoir for this virus strain in the Mediterranean basin, as B. tabaci Mediterranean (MED), the most prevalent species of the complex in the region, is not an efficient vector of this begomovirus between cultivated zucchini and wild E. elaterium plants.

Metabolic engineering of cucurbitacins in Cucurbita pepo hairy roots.

In this paper we show that metabolic engineering in Cucurbita pepo hairy roots can be used to both effectively increase and modify cucurbitacins. Cucurbitacins are highly-oxygenated triterpenoids originally described in the Cucurbitaceae family, but have since been found in 15 taxonomically distant plant families. Cucurbitacin B, D, E and I are the most widespread amongst the Cucurbitaceae and they have both important biological and pharmacological activities. In this study C. pepo hairy roots were used as a platform to boost production and alter the structures of the afore mentioned cucurbitacins by metabolic engineering to potentially provide new or more desirable bioactivities. We report that the ability to induce cucurbitacin biosynthesis by basic Helix-Loop-Helix transcription factors is partially conserved within the Cucurbitaceae and therefore can potentially be used as a biotechnological tool to increase cucurbitacins in several genera of this family. Additionally, overexpression of a novel acyltransferase from cucurbitacin producing Iberis amara generates a hitherto undescribed acetylation at the C3-hydroxyl group of the cucurbitadienol backbone. While overexpression of the cytochromes P450 CsCYP88L2 and McCYP88L7 from Cucumis sativus and Momordica charantia (respectively), results in accumulation of new spectral feature as revealed by High resolution liquid chromatography mass spectroscopy analysis; the m/z of the new peak supports it might be a cucurbitacin hydroxylated at the C19 position in C. pepo hairy roots. Finally, this paper is a case study of how hairy roots can be used to metabolically engineer and introduce novel modifications in metabolic pathways that have not been fully elucidated.

Ecballium elaterium improved stimulatory effects of tissue-resident NK cells and ameliorated liver fibrosis in a thioacetamide mice model.

Ecballium elaterium (EE), widely used plant in Mediterranean medicine, showed anticancer activity. This study aimed to investigate EE effects on liver fibrosis in an animal model of thioacetamide (TAA). Intraperitoneal administration of TAA was performed twice weekly for four weeks in C57BL6J mice. Livers were extracted and serum were evaluated for inflammatory markers (H&E staining, ALT, AST, ALP), pro-inflammatory cytokines, fibrosis (Sirius red staining, Masson's trichrome, α-smooth muscle actin and collagen III), and metabolic (cholesterol, triglyceride, C-peptide, and fasting-blood-sugar) profiles. Glutathione, glutathione peroxidase, and catalase liver antioxidant markers were assessed. Tissue-resident NK cells from mice livers were functionally assessed for activating receptors and cytotoxicity. Compared to vehicle-treated mice, the TAA-induced liver injury showed attenuation in the histopathology outcome following EE treatment. In addition, EE-treated mice resulted in decreased serum levels of ALT, AST, and ALP, associated with a decrease in IL-20, TGF-ß, IL-17, IL-22 and MCP-1 concentrations. Moreover, EE-treated mice exhibited improved lipid profile of cholesterol, triglycerides, C-peptide, and FBS. EE treatment maintained GSH, GPX, and CAT liver antioxidant activity and led to elevated counts of tissue-resident NK (trNK) cells in the TAA-mice. Consequently, trNK demonstrated an increase in CD107a and IFN-γ with improved potentials to kill activated hepatic-stellate cells in an in vitro assay. EE exhibited antifibrotic and antioxidative effects, increased the number of trNK cells, and improved metabolic outcomes. This plant extract could be a targeted therapy for patients with advanced liver injury.

Ecballium elaterium attenuates neuroinflammation in an animal model of Alzheimer's disease through modulation of nuclear factor κB pathway.

OBJECTIVE: Sustained inflammation, which could be promoted by Aß aggregation and tau hyperphosphorylation, is a critical player in Alzheimer's disease (AD) pathogenesis. In the first phase, this study was designed to evaluate the anti-inflammatory properties of Ecballium elaterium (EE), as a Mediterranean therapeutic plant, and its effects on biochemical and behavioral signs of nucleus basalis of Meynert lesioned (NBML) rats, as an approved model of AD. In the second phase, we investigated the effect of EE on nuclear factor (NF)-κB pathway which is responsible for encoding proteins involved in the inflammatory cascade. MATERIALS AND METHODS: Animals were divided randomly into four groups as following: control, NBML rats (AD), AD rats that were treated by high- and low-dose EE. Prostaglandins (PGs) levels were measured by enzyme-linked immunosorbent assay (ELISA) kits. Cyclooxygenase-2 (COX-2) and acetylcholinesterase (AChE) levels were assessed by fluorometric kit and Elman method, respectively. Behavioral signs were evaluated by Morris Water Maze (MWM) test and inflammatory proteins content was analyzed by immunoblotting method. RESULTS: According to the results, treatment of NBML rats with EE fruit juice reduced PGs and cytokines more than 2-fold in comparison with AD rats through inhibition of COX-2 enzyme. Attenuation of inflammatory response in NBML rats was accompanied by reduced AChE activity (about 3-fold) and improved learning ability. Interestingly, EE reduced NF-κB expression for about 3-fold which resulted in a more than 10-fold increase in IκBα/P-IκBα ratio. CONCLUSION: Our results confirmed the TNF-α/cytokines/NF-κB/COX-2 pathway involves as the main inflammatory response in NBML rats. We also provided biochemical and behavioral evidence which introduces EE as an anti-inflammatory adjuvant to improve pathophysiological signs in patients suffering from AD and related dementia.

Investigation of phytochemical contents, in vitro antioxidant and antibacterial behavior and in vivo anti-inflammatory potential of Ecballium elaterium methanol fruits extract

Abstract Ecballium elaterium species are mostly used as therapeutic agents and food ingredient. The current work was designed to investigate phytochemical contents, antioxidant, antibacterial, and anti-inflammatory properties of methanol fruits extract of Ecballium elaterium. Good antioxidant activity was observed with IC50 values of 156 ± 4 and 377 ± 6 μg/mL for DPPH and ABTS, respectively, and EC50 of 126 ± 4 µg/mL for FRAP assays, which is related with their richness in total phenolic, flavonoid and condensed tannins contents. The results of antibacterial activity showed the effectiveness of methanol extract against Bacillus cereus with value of inhibition zone diameter of 15 ± 0 mm and a MIC and MBC values of 6 ± 0 and 12 ± 0 mg/mL, respectively. The in vivo anti-inflammatory effects have been also studied by carrageenan induced rat paw edema assay and the results revealed that a dose of 75 mg/kg induced a significant inhibition of 66.4% at 2 h. FT-IR spectral data justified the presence of biological functional groups such as ─OH, C─H, C─O, C─C and C=O. These results highlighted the potential using of Ecballium elaterium fruits extract as natural antimicrobial, antioxidant and anti-inflammatory agents for food applications and for the pharmaceutical industry.

Effects of Ecballium elaterium on brain in a rat model of sepsis-associated encephalopathy.

Despite recent advances in antibiotic therapy, sepsis remains a major clinical challenge in intensive care units. Here we examined the anti-inflammatory and antioxidant effects of Ecballium elaterium (EE) on brain, and explored its therapeutic potential in an animal model of sepsis-associated encephalopathy (SAE) [induced by cecal ligation and puncture (CLP)]. Thirty rats were divided into three groups of 10 each: control, sepsis, and treatment. Rats were subjected to CLP except for the control group, which underwent laparatomy only. The treatment group received 2.5 mg/kg EE while the sepsis group was administered by saline. Twenty-four hours after laparotomy, animals were sacrificied and the brains were removed. Brain homogenates were prepared to assess interleukin 1beta (IL-1ß), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), total antioxidant capacity (TAC), and total oxidant status (TOS). Brain tissue sections were stained by hematoxylin and eosin (H&E) to semi-quantitatively examine the histopathologic changes such as neuron degeneration, pericellular/perivascular edema and inflammatory cell infiltration in the cerebral cortex. We found a statistically significant reduction in brain tissue homogenate levels of TNF-α 59.5 ± 8.4/50.2 ± 6.2 (p = 0.007) and TOS 99.3 ± 16.9/82.3 ± 7.8 (p = 0.01) in rats treated with EE; although interleukin 6 levels were increased in the treatment group compared to the sepsis group, this was not statistically significant. Neuronal damage (p = 0.00), pericellular/perivascular edema and inflammatory cell infiltration (p = 0.001) were also significantly lower in the treatment group compared to those in the sepsis group. These data suggest that Ecballium elaterium contains some components that exert protective effects against SAE in part by attenuating accumulation of proinflammatory cytokines, which may be important contributors to its anti-inflammatory effects during sepsis.

Purification of Cucurbitacins D, E, and I from Ecballium Elaterium (L.) A. Rich Fruits and Study of Their Cytotoxic Effects on the AGS Cell Line

Background: The plant Ecballium elaterium (L.) A. Rich, belongs to the Cucurbitaceae family which occupies an important position in traditional medicine prescriptions. It has been reported that a freeze-dried aqueous extract of E. elaterium fruits has cytotoxic effects on the AGS human stomach adenocarcinoma cell line. We here focused on anticancer effects of the main chemicals purified from E. elaterium fruits. Materials and Methods: We isolated cucurbitacins D, E, and I from chloroform, and ethyl acetate fractions of a methanolic extract of E. elaterium fruits and assessed their cytotoxic effects on the AGS cell line by MTT assay. The methanolic extract was fractionated to petroleum ether, chloroform, and ethyl acetate fractions. The compounds isolated by column chromatography were identified by NMR spectroscopy. Results: After 24 h of incubation with AGS cells, the IC50 values were 0.3, 0.1, and 0.5 µg/ml for cucurbitacins D, E, and I respectively. Conclusions: This finding suggests that because of its cucurbitacins, E. elaterium fruit may have some cytotoxic effects on gastric cancer cells. Also, compared with D and I, cucurbitacin E showed greater potency in this regard.

Targetting αvß3 and α5ß1 integrins with Ecballium elaterium (L.) A. Rich. seed oil.

In the present study, the effect of Ecbalium elaterium seed oil on adhesion, migration and proliferation of human brain cancer cell line (U87) was determined. Treatment of U87 cell line with the seed oil resulted in strong inhibition of their adhesion to fibrinogen (Fg), fibronectin (Fn). It also reduced their migration and proliferation in a dose-dependent manner without being cytotoxic. Concomitantly, by using Matrigel™ assays, the oil significantly inhibited angiogenesis. The anti- tumor effect of the oil is specifically mediated by αvß3 and α5ß1 integrins. The presence of integrin antagonists in seed oil from E. elaterium could be used for the development of anticancer drugs with targeted "multi-modal" therapies combining anti-adhesif, antiproliferative, antimetastasic and anti-angiogenic, approaches.

Effects of Ecballium Elaterium on Proinflammatory Cytokines in a Rat Model of Sepsis.

OBJECTIVE: Ecballium elaterium (EE) is a plant from Cucurbitaceae family. Its anti-inflammatory role in sepsis is not well understood. We investigated the effects of EE on serum levels of proinflammatory cytokines and further explored the mechanisms underlying histological changes in liver and ileum following EE administration in a polymicrobial sepsis model. METHODS: Thirty rats were divided into three groups of 10 rats each. Rats were subjected to sham laparotomy plus normal saline administration (control group, CG), laparotomy with cecal ligation and puncture (CLP) (sepsis group, SG), and laparotomy with CLP plus 2.5 mg/kg EE administration (experimental group, EG). Twenty-four hours after laparotomy, animals underwent cardiac puncture, and blood was collected for interleukin 1 (IL-1), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) assessment. Whole sections of liver and ileum tissues were collected for histologic examination. RESULTS: The serum level of IL-6 was significantly lower in EG as compared to SG. Although IL-6 levels were shown a statistically significant (p < 0.0001) decline to near control values, no significant changes were observed in serum levels of IL-1 and TNF-α after EE treatment. Histologic examination revealed statistically significant reduction in collagen formation (p = 0.001) on serosal surface of ileum and hepatic venous congestion (p = 0.040) in EG as compared to SG. CONCLUSION: EE might play a protective role in sepsis prevention and treatment by decreasing IL-6 production and reducing liver damage and may influence bacterial translocation by reinforcing intestinal barrier function.

Histopathological and Biochemical Effects of Ecballium elaterium on Sepsis-Induced Lung Injury.

PURPOSE: The aim of this study was to investigate the role of Ecballium elaterium (EE) on sepsis-induced lung injury. MATERIALS AND METHODS: A total of 30 male rats were divided into three groups as follows: control, sepsis, and treatment groups (sepsis + EE) with each group containing 10 rats. A rat model of sepsis induced by cecal ligation and puncture (CLP) was used. In the treatment group, rats were injected intraperitoneally with 2.5 mg/kg EE after CLP. Interleukin-6 (IL-6), tumor necrosis factor (TNF)-α, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) values after a 24-hr period were measured via cardiac puncture. Animals were harvested after the procedure and biochemical analysis was done and histopathological changes of the tissue sections of lungs were examined thereafter. RESULTS: A statistically significant decrease was observed in the IL-6 (p < .05), TNF-α (p < .05), and TOS (p < .01) levels in the sera of the treatment group compared to those of the sepsis group. Following the treatment, the TOS (p = .01) and OSI (p < .05) levels in the lung tissue of rats indicated a statistically significant decrease compared to those of the sepsis group. The histopathological follow-up undertaken after the administration of the EE treatment to septic rats showed significantly lower values of alveolar wall thickness (p < .001), interstitial edema (p = .018), and neutrophil infiltration (p = .047). CONCLUSION: EE treatment may have beneficial effects on sepsis-induced lung injury, and therefore has potential for clinical use.

A new experimental rat model of pancreatitis using Ecballium elaterium.

BACKGROUND: We originally aimed to determine the beneficial effects of Ecballium Elaterium (EE) on acute pancreatitis; however, we observed negative effects of EE on the pancreas. Thus, we used EE in rats to generate a new model of pancreatitis, which we compared with other established pancreatitis models. METHODS: A total of 32 Wistar albino rats were used. Rats were divided into 4 groups, each of which contained 8 rats. Group 1 (Control), Group 2 (L-Arginine (LA), Group 3 (LA + EE), Group 4 (EE): Twenty-four hours after that serum parameters were analyzed in the collected blood. Blood samples were transported on mice to the Biochemistry Laboratory following cardiac puncture. The levels of amylase, interleukin (IL)-6, interleukin (IL) 1-ß (IL-1ß), malondialdhyde (MDA), tumor necrosis factor (TNF)-α, total antioxidant status (TAS), and total oxidant status (TOS) were analyzed. Histopathological analysis: The pancreas and lung tissue samples obtained from the rats. Edema, inflammation, vacuolization, and necrosis of the pancreas were assessed using a scoring system ranging from 0 to 4. Edema, hemorrhage and inflammation of the lung tissue were evaluated using a scoring system ranging from 0 to 3. RESULTS: Histopathological analysis revealed that edema, inflammation, necrosis, and hemorrhage were significantly higher in the LA + EE group than in the control group. Moreover, necrosis was higher in the rats that received LA and EE compared to the rats that received only LA or EE. Increases in inflammatory mediator levels, including IL-6, IL-1ß, TNF-α, MDA, and TOS, were observed in all groups as compared to the control group. Moreover, lower TAS levels were detected in all groups but the control group. The increase in IL-1ß and TNF-α levels and the decrease in TAS were statistically significant in all groups (P < 0.05). CONCLUSIONS: EE may be used to create a successful acute pancreatitis (AP) model, resulting in edema, necrosis, hemorrhage, and inflammation of the pancreas. The major advantage of this model is that it does not require laparotomy, and can be implemented with only an intraperitoneal injection (IP). Moreover, EE may be combined with other agents, such as LA, to create severe pancreatitis. Further molecular studies are warranted to determine the underlying effects of EE on the pancreas.

Hepatoprotective and proapoptotic effect of Ecballium elaterium on CCl4-induced hepatotoxicity in rats.

OBJECTIVES: To investigate the effects of perorally administered juice on tetrachloromethane (CCl4)-induced hepatotoxicity model in rats. METHODS: Male Wistar rats were tube-administrated silymarin, Ecballium juice at 0.2 mL/kg and 0.7 mL/kg daily for 3 consequent days, i.e., 3.28 µg and 11.48 µg of cucurbitacin B per kg of body weight respectively. On the third day, liver damage was induced by intraperitoneal application of CCl4. On the fourth day, abdominal cavity was macroscopically examined and liver samples were taken for histopathological and immunochemical evaluation. HPLC was used to determine the content of the active substance cucurbitacin B. RESULTS: The experiment revealed that 0.7 ml/kg juice concentration expressed the highest pro-apoptotic activity, but with prevailing negative effects. Compared with the lower concentration, there was an observable vasodilatation with consequent interstitial hemorrhages and a larger scope of inflammatory damage, which suppressed the hepatoprotective effect. In the 0.2 mL/kg concentration, there was a smaller pro-apoptotic activity but other parameters had better results, and the liver parenchyma damage was reversible. CONCLUSIONS: No reactions confirming the potentially allergic effect on laboratory rats were observed; its hepatoprotective and anti-inflammatory effect was confirmed on a model of acute liver damage.

Severe uvular edema and resulting hypoxemia due to single use of Ecballium elaterium extract.

BACKGROUND: Ecballium elaterium (also known as squirting cucumber in the Mediterranean countries) is a weedy plant that belongs to the Cucurbitacea family. The roots and juice of its fruits were used traditionally against a wide range of diseases and symptoms, since it was known for its anti-inflammatory and analgesic effects. The use of the extract, though, has been linked to allergic reactions and toxic effects. In this report, we present a case of severe uvular edema and hypoxemia due to a single use of Ecballium elaterium extract, and we discuss the possibility of life-threatening conditions resulting from the utilization of this particular plant as a remedy. CASE REPORT: A male Caucasian presented to the ER with complaints of troubled breathing and retrosternal discomfort that started a few hours before arriving at the ER. The patient's medical history revealed the use of Ecballium elaterium extract. Clinical examination confirmed severe uvular edema and hypoxemia. Administration of oxygen and IV drugs (Hydrocortisone) relieved the symptoms and improved the patient's condition. After a 24-hour hospitalization the patient was discharged without further complaints. CONCLUSIONS: The use of Ecballium elaterium extract as a remedy can cause severe adverse effects that should not be ignored. Uvular edema is one of the most common and can lead, as shown above, to hypoxemia, which is a potentially life-threatening condition.

Effect of ethanolic extract of Ecballium elaterium against Staphylococcus aureus and Candida albicans.

OBJECTIVE: To evaluate the antimicrobial activity of ethanolic extract of Ecballium elaterium (E. elaterium) fruits alone against Staphylococcus aureus (S. aureus) strains and Candida albicans (C. albicans) strains, or in combination with penicillin against Staphylococcus areus strains. METHODS: Evaluation of the antimicrobial activity or synergy interaction was carried out using microdilution method. RESULTS: The results showed that ethanolic extract of E. elaterium fruits has antimicrobial activity against methicillin resistant S. aureus (MRSA), methicillin sensitive S. aureus (MSSA) and C. albicans. This extract showed a significant decrease in minimum inhibitory concentrations (MIC) of penicillin against both MRSA and MSSA strains. Fractional inhibitory concentration index (FIC) between penicillin and ethanolic extract of E. elaterium fruits against these test strains was less than 0.5. CONCLUSIONS: This study suggests that ethanolic extract of E. elaterium fruits has antimicrobial activity against S. aureus and C. albicans and there is a possibility of concurrent use of penicillin and E. elaterium extract in combination in the treatment of infections caused by MRSA and MSSA strains. A wider study is needed to identify the effective components, the mode of action and the possible toxic effect in vivo of these ingredients.

Foraging behavior of three bee species in a natural mimicry system: female flowers which mimic male flowers in Ecballium elaterium (Cucurbitaceae).

The behavior of Apis mellifera and two species of solitary bees which forage in the flowers of monoecious Ecballium elaterium (L.) A. Rich (Cucurbitaceae) were compared. The female flowers of E. elaterium resemble male flowers visually but are nectarless, and their number is relatively smaller. Apis mellifera was found to discriminate between the two genders and to pay relatively fewer visits to female flowers (mean of 30% relative to male flowers) from the beginning of their activity in the morning. The time spent by honeybees in female flowers is very short compared to that spent in male flowers. It is surmised that the bees remember the differences between the flowers where they foraged on the previous days. In contrast, the two species of solitary bees Lasioglossum politum (Morawitz) (Halictidae) and Ceratina mandibularis Fiese (Anthophoridae) visit the female flowers with nearly equal frequencies at the beginning of each foraging day and stay longer in these flowers. Over the day there is a decline in the relative frequency of visits to female flowers and also in the mean time spent in them. The study shows that bees can collect rewards at high efficiency from the flowers of Ecballium elaterium because of their partial discrimination ability and the scarcity of the mimic flowers. It is suggested that the memory pattern of some solitary bees may be different from that of Apis mellifera. It seems that the limited memory and discrimination ability of bees can lead to a high frequency of visits to the mimic flowers during a long flowering season.