Antidiabetic properties of aqueous and methanol extracts from the trunk bark of Ceiba pentandra in type 2 diabetic rat

The type 2 diabetes is one of the major global health issues that affects millions of people. This study evaluated the antidiabetic activity of aqueous extracts (AECP) and methanol extracts (MECP) from Ceiba pentandra trunk bark on an experimental model of type 2 diabetes (T2D). This model was induced in rats by the combination of a high-fat diet (HFD) and a single dose of streptozotocin (40 mg/kg, intraperitoneal) at the seventh day of experimentation. Diabetes was confirmed on day 10 by fasting blood glucose more than or equal to 200 mg/dL. Diabetic animals still under HFD were treated orally and twice daily, with MECP and AECP (75 and 150 mg/kg) or metformin (40 mg/kg) for 14 days. During the experiment, blood glucose and animal weights were determined. Oral glucose tolerance test was performed on day 15, followed by animals sacrifice for blood, liver, and pancreas collection. Total cholesterol and triglyceride levels were evaluated in plasma, whereas malondialdehyde (MDA), glutathione (GSH), superoxide dismutase, and catalase were quantified in tissue homogenates. AECP and MECP significantly reduced the hyperglycemia by up to 62% and significantly improved the oral glucose tolerance test. The impaired levels of cholesterol and triglycerides registered in diabetic control were significantly reversed by both extracts at all the doses used. Alterations in diabetic pancreas weight, GSH, and MDA were also significantly reversed by plant extracts. AECP and MECP possess type 2 antidiabetic effects that could result from their ability to improve the peripheral use of glucose, lipid metabolism or from their capacity to reduce oxidative stress. These finding provide a new avenue for better control and management of early or advanced T2D.     

Experimental study on the antioxidant activity of Malus hupehensis (Pamp.) Rehd extracts in vitro and in vivo

Extracts of Malus hupehensis (Pamp.) Rehder, containing flavonoids with good antioxidant and antiliver injury properties, possess various biological activities. The aim of this study was to explore the antioxidant activity of these extracts in vitro and in vivo. The antioxidant activity of the extracts was studied using scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, and superoxide free radicals and by inhibiting mushroom tyrosinase activity in vitro. An in vivo antioxidant experiment was performed using a rat-aging model. Aging was induced in rats with D-galactose through treating them at doses of extracts about 150, 300, and 600 mg·kg⁻¹·day⁻¹. The Malus hupehensis extracts showed high antioxidant activity; the IC₅₀ values of DPPH radicals, ABTS radicals, superoxide radicals, and mushroom tyrosinase inhibition were 19.00 μg/mL, 303.94 μg/mL, and 3.71 mg/mL, and 1.16 mg/mL, respectively. Our experiments showed that the extracts significantly increased the activity of antioxidant enzymes in the serum and tissue homogenate in vivo, and that the effects were positively correlated with the dose, compared with the activity observed in controls. Histopathological observation also confirmed that the extracts had protective effects after oxidative injury in rat tissues. In conclusion, the extracts of M. hupehensis showed effective antioxidant activity both in vitro and in vivo.     

Ethyl acetate extracts of alfalfa (Medicago sativa L.) sprouts inhibit lipopolysaccharide-induced inflammation in vitro and in vivo

This study aimed to investigate if food components that exert anti-inflammatory effects may be used for inflammatory disorders by examining alfalfa sprout ethyl acetate extract (ASEA). The cytokine profile and life span of BALB/c mice with acute inflammation after intra-peritoneal (ip) injection of 15 mg/kg BW lipopolysaccharide (LPS) were determined. The results showed that the life span of LPS-induced inflammatory mice were negatively correlated with serum levels of TNF-α, IL-6, and IL-1β at 9 hr after LPS-injection, which indicated that suppressing these cytokines in the late phase of inflammation may be beneficial for survival. The in vitro experiment then showed that ASEA significantly reduced IL-6 and IL-1β production and the NF-κB trans-activation activity of mitogen-stimulated RAW264.7 cells. To further evaluate the anti-inflammatory effects of ASEA in vivo, BALB/c mice were tube-fed with 25 mg ASEA/kg BW/day in 50 μl sunflower oil, while the control and PDTC (pyrrolidine dithiocarbamate, an anti-inflammatory agent) groups were tube-fed with 50 μl sunflower oil/day only. After one week of tube-feeding, the PDTC group was injected with 50 mg/kg BW PDTC and one hour later, all of the mice were injected with 15 mg/kg BW LPS. The results showed that the ASEA and PDTC groups had significantly lower serum TNF-α, IL-6, and IL-1β levels at 9 hr after LPS challenge, and significantly higher survival rates than the control group. This study suggests that ASEA supplementation can suppress the production of pro-inflammatory cytokines and alleviate acute inflammatory hazards.     

Extreme long-distance dispersal of the lowland tropical rainforest tree Ceiba pentandra L. (Malvaceae) in Africa and the Neotropics

Many tropical tree species occupy continental expanses of rainforest and flank dispersal barriers such as oceans and mountains. The role of long-distance dispersal in establishing the range of such species is poorly understood. In this study, we test vicariance hypotheses for range disjunctions in the rainforest tree Ceiba pentandra, which is naturally widespread across equatorial Africa and the Neotropics. Approximate molecular clocks were applied to nuclear ribosomal [ITS (internal transcribed spacer)] and chloroplast (psbB-psbF) spacer DNA sampled from 12 Neotropical and five West African populations. The ITS (N=5) and psbB-psbF (N=2) haplotypes exhibited few nucleotide differences, and ITS and psbB-psbF haplotypes were shared by populations on both continents. The low levels of nucleotide divergence falsify vicariance explanations for transatlantic and cross-Andean range disjunctions. The study shows how extreme long-distance dispersal, via wind or marine currents, creates taxonomic similarities in the plant communities of Africa and the Neotropics.     

Lenalidomide enhances antibody-dependent cellular cytotoxicity of solid tumor cells in vitro: influence of host immune and tumor markers

We evaluated the effect of combining lenalidomide with therapeutic antibodies on antibody-dependant cell-mediated cytotoxicity (ADCC) of solid tumor cells, and the requirement for expression of natural killer (NK) cell-activating receptors and their solid tumor surface ligands. Twenty-three human tumor cell lines (colon, breast, lung, head and neck, ovary, and bone sarcoma) were analyzed. NK effector cells were isolated from healthy donors, pre-treated with and without lenalidomide, and incubated with antibody-coated tumor cells to determine ADCC. In blocking experiments, NK cells were pre-incubated with anti-DNAM-1 or anti-NKG2D antibodies, and target colorectal cells were pre-incubated with anti-CD155 (PVR), anti-MIC-A/B, or anti-ULBP 3 antibodies. Differences between groups were assessed using unpaired and paired Student’s t test and one-way ANOVA. Lenalidomide enhanced NK cell-mediated ADCC of trastuzumab- and cetuximab-coated tumor cells. Activity against colorectal cancer cells was dependent on target antigen expression, but independent of KRAS status and FcγRIIIa genotype. The extent of ADCC and its enhancement by lenalidomide correlated with NK cell expression of NKG2D and DNAM-1, and tumor cell expression of PVR and MIC-A. Blocking of NKG2D and, to a lesser extent, DNAM-1 inhibited ADCC. Anti-MIC-A/B monoclonal antibody blocked natural cytotoxicity, but not ADCC. Lenalidomide enhances the ability of IgG1-isotype antibodies to mediate ADCC of solid tumor cells, the extent of which is largely dependent on NKG2D–NKG2D ligand interactions, but appears to be independent of MIC-A/B. This provides a rationale for exploratory clinical studies and an assessment of potential biomarkers predictive of clinical benefit.     

Proliferative activity of Echinacea angustifolia root extracts on cancer cells: Interference with doxorubicin cytotoxicity

Doxorubicin is an anticancer drug that causes apoptosis in cells, but cardiotoxicity limits the cumulative dose that can remain in the blood. Echinacea extracts have been prescribed to supplement cancer chemotherapy. In a recent study, it was reported that Echinacea purpurea extracts protected noncancerous cells from apoptosis. Our study aimed to determine interference with doxorubicin chemotherapy, and if fractions and compounds from Echinacea angustifolia roots protected the cells. Cervical and breast cancer cells were treated with the Echinacea samples and doxorubicin. At 0.05 and 0.5 microM doxorubicin concentration, cynarine increased HeLa cell growth by 48-125% and 29-101%, respectively (p<0.01). At 0.05 microM doxorubicin concentration, chicoric acid increased cell growth by 23-100% (p<0.01). When MCF-7 cells were treated with Echinacea and doxorubicin, the ethyl acetate fraction increased cell growth by 20-25%, and chicoric acid increased cell growth by 10-15%. Cynarine showed proliferative activity on HeLa cells, but showed antiproliferative activity on MCF-7 cells. Results indicate that phenolic compounds are responsible for proliferative activity. Studies with individual compounds show that chicoric acid and cynarine interfered with cells treated with 0.5 microM doxorubicin. The results of this study show that Echinacea herbal medicines affect cell proliferation despite cancer treatment, and that herbal medicines require further study with respect to anticancer drugs.     

Chemiluminescence,suppression and cytotoxicity in peripheral blood mononuclear cells from solid tumor cancer patients

Summary Chemiluminescence, indomethacin-sensitive suppression, and adherent cell cytotoxicity were measured in peripheral blood mononuclear cells (PBMC) from normal subjects and solid tumor cancer patients. These functions were found to be differentially affected by malignant disease. In cancer patients with disseminated disease, indomethacin-sensitive suppression and chemiluminescence emission were increased to a level significantly higher than normal without a concurrent increase in adherent cell cytotoxic function. In cancer patients with at most minimum residual diseases, the levels of chemiluminescence, indomethacin-sensitive suppression, and adherent cell cytotoxicity found were comparable to those of the normal study population. In vitro stimulation of cells from patients with disseminated disease by phorbol myristic acetate (PMA) increased chemiluminescence overcame the suppressive effects of indomethacin-sensitive suppressor cells, and increased adherent cell cytotoxicity; in cells from patients with at most minimum residual disease, PMA increased chemiluminescence and cytotoxicity without influencing the activity of indomethacin-sensitive suppressor cells. Vaccination of lung cancer patients with Freund's complete adjuvant or Freund's complete adjuvant plus tumor antigen extracts led to increased levels of chemiluminescence and increased levels of adherent cell cytotoxicity without altering indomethacin-sensitive regulatory cell function.     

Evaluation of a gene-directed enzyme-product therapy (GDEPT) in human pancreatic tumor cells and their use as in vivo models for pancreatic cancer

Background

Gene-directed enzyme prodrug therapy (GDEPT) is a two-step treatment protocol for solid tumors that involves the transfer of a gene encoding a prodrug-activating enzyme followed by administration of the inactive prodrug that is subsequently activated by the enzyme to its tumor toxic form. However, the establishment of such novel treatment regimes to combat pancreatic cancer requires defined and robust animal model systems.

Methods

Here, we comprehensively compared six human pancreatic cancer cell lines (PaCa-44, PANC-1, MIA PaCa-2, Hs-766T, Capan-2, and BxPc-3) in subcutaneous and orthotopical mouse models as well as in their susceptibility to different GDEPTs.

Results

Tumor uptake was 83% to 100% in the subcutaneous model and 60% to 100% in the orthotopical mouse model, except for Hs-766T cells, which did not grow orthotopically. Pathohistological analyses of the orthotopical models revealed an infiltrative growth of almost all tumors into the pancreas; however, the different cell lines gave rise to tumors with different morphological characteristics. All of the resultant tumors were positive for MUC-1 staining indicating their origin from glandular or ductal epithelium, but revealed scattered pan-cytokeratin staining. Transfer of the cytochrome P450 and cytosine deaminase suicide gene, respectively, into the pancreatic cancer cell lines using retroviral vector technology revealed high level infectibility of these cell lines and allowed the analysis of the sensitivity of these cells to the chemotherapeutic drugs ifosfamide and 5-fluorocytosine, respectively.

Conclusion

These data qualify the cell lines as part of valuable in vitro and in vivo models for the use in defined preclinical studies for pancreas tumor therapy.     

Nitrate reductase activity (in vivo and in vitro) of ditelosomic stocks of wheat (Triticum aestivum L.)

The nitrate reductase activities (NRA) of 31 ditelosomic stocks were compared with that of the control plant [Chinese Spring (CS) euploid], using in vivo and in vitro assay procedures that had been optimized with respect to the euploid. Fourteen stocks exhibited significant differences in in vivo NRA from that of the euploid; the effect of removal of a chromosome arm was always to increase NRA. Eight of these stocks showed similar effects in vitro, although in three, a casein-sensitive factor had to be eliminated before the difference was expressed. Homoeologous group effects were evident among ditelosomics of groups 2, 4, and 7, while for three chromosomes (2D, 7A, and 7B), removal of either arm resulted in a similar increase in NRA in vivo and probably in vitro.P. W. Jones was supported by a Science Research Council C.A.S.E. award with the Plant Breeding Institute, Cambridge, U.K.     

Slow cytotoxicity of the polysaccharide levan on tumor cells in vitro

Previous studies have shown that 1 h preincubation with levan caused a sharp decrease in tumorigenicity of tumor cells, although cell viability was not affected. In the present study, the possibility that levan might change the sensitivity of Lewis lung carcinoma cells to the host immune system and that levan might cause delayed damage were tested. Cell morphology, DNA synthesis, cell multiplication and viability as well as osmotic fragility were followed during several days in culture. Levan was found not to affect cell immunogenicity. The polysaccharide destroyed tumor cells by a rather peculiar mechanism: after a seemingly normal or even enhanced growth in culture during 3-5 days, cells suddenly burst. During the apparently normal growth, several morphological changes were observed: cell volume increased, cytoplasm swelled by apparent water uptake and some cells contained two or more nuclei. Levan-treated cells were found to be more susceptible to osmotic shock than non-treated cells. The reason for the sudden cell death could be a gradual increase in volume, up to a point which is no more compatible with membrane integrity, resulting in cell lysis.     

Effect of liquid seaweed extracts on growth of tomato seedlings (Solanum lycopersicum L.)

Seaweed extracts are used as nutrient supplements, biostimulants, or biofertilizers in agriculture and horticulture to increase plant growth and yield. In this study, we examined the effect of liquid seaweed extracts (LSEs) made from Ulva lactuca, Caulerpa sertularioides, Padina gymnospora, and Sargassum liebmannii as biostimulants on the germination and growth of tomato (Solanum lycopersicum) under laboratory and greenhouse conditions using foliar and soil drench applications of LSEs. We assessed LSEs at different concentrations (0.2, 0.4, and 1.0 %) on germination parameters (percentage, index, mean time, energy, and seedling vigor index) and growth parameters (plumule length, radical length, shoot length, root length, fresh weight, and dry weight) of tomato seedlings. Our results indicate that seeds treated with LSEs of U. lactuca and P. gymnospora at lower concentrations (0.2 %) showed enhanced germination (better response in germination rate associated with lower mean germination time, high germination index and germination energy, and consequently greater seedling vigor and greater plumule and radicle length). Application as a soil drench was found to be more effective in influencing the height of the plant (up to 79 cm) than the foliar spray application (75 cm). Plants receiving LSEs of U. lactuca and P. gymnospora showed increased shoot length, root length, and weight. Furthermore, U. lactuca and P. gymnospora were found to be more successful and better candidates for developing effective biostimulants to improve the growth of tomato plants. This study provides important information on the identification and utilization of Mexican seaweed resources for agriculture and is the first study to report on the uses of these seaweeds as a source of liquid extracts as biostimulants in agriculture.     

Relationship between in vitro tumor stem cell assay and in vivo antitumor activity using the P388 leukemia

The relationship between in vitro tumor stem cell sensitivity and in vivo antitumor efficacy using 13 active antineoplastic agents was examined by means of the intraperitoneal P388 mouse leukemia system. The doses, which produced a 50% increase in life span after one, five and nine days of treatment, were all significantly correlated with the in vitro concentration producing a 70% reduction in colony formation during a seven-day continuous drug exposure. The five-day correlation was the best, followed by nine days and one day. Correlations were not improved by corrections for either in vitro drug stability or toxicity (LD50). The highly significant correlations observed in this simple retrospective analysis provide a basis for the development of more sophisticated models for the prediction of in vivo results from in vitro data.     

The antitumor activity of liposomal aclarubicin in vitro and in vivo]

Study on antitumor activity of free and liposomal anthracycline antibiotic aclarubicin in vitro and in vivo showed that liposomal aclarubicin was characterised by activity against ascitic Ehrlich carcinoma comparable to that of free aclarubicin when used in a dose of 25 mg/kg. Liposomal antibiotic had a more pronounced antimetastatic action and showed no toxicity (in a dose of 30 mg/kg). Liposomal aclarubicin had a higher activating capacity with respect to the macrophage tumoricidal properties.     

Synthesis,cytotoxicity, and in vivo antitumor activity study of parthenolide semicarbazones and thiosemicarbazones

Parthenolide is an important sesquiterpene lactone with potent anticancer activities. In order to further improve its biological activity, a series of parthenolide semicarbazone or thiosemicarbazone derivatives was synthesized and evaluated for their anticancer activity. Derivatives were tested in vitro against 5 human tumor cell lines, and many of these showed higher cytotoxicity than parthenolide. Five compounds were further studied for their antitumor activity in mice. The in vivo result indicated that compound 4d showed both promising antitumor activity against mice colon tumor and small side effects on immune systems. The cell apoptosis and cell cycle distribution of compound 4d were also studied. Molecular docking studies revealed multiple interactions between 4d and NF-κB. Our findings demonstrate the potential of semicarbazones as a promising type of compounds with anticancer activity.     

Steroidal constituents and anti-inflammatory activity of the horse chestnut (Aesculus hippocastanum L.) bark

The content and the composition of sterols in the bark of the horse chestnut, Aesculus hippocastanum L., have been examined. Stigmasterol and alpha-spinasterol were, with the beta-sitosterol, the most abundant sterols. The petrol extract from bark shows anti-inflammatory activity.     

Effects of aloe extracts on human normal and tumor cells in vitro

Fractions of leaf extracts from 2 local types, labeledAloe vera (subsequently identified asAloe barbadensis Mill, andA. saponaria Haw.), were prepared by differential centrifugation and tested by in vitro assays for the presence of lectinlike activities and for effects on the attachment and growth of human normal and tumor cells. Fractions of extracts of fresh leaves and commercially “stabilized”Aloe vera gel had high levels of lectin-like substances measured by immunodiffusion and hemagglutination assays. Substances in fluid fractions from both fresh leaf sources were found to markedly promote attachment and growth of human normal, but not tumor, cells and to enhance healing of wounded cell monolayers. In contrast, fractions of “stabilized”Aloe vera gel were equally cytotoxic for human normal and tumor cells in vitro. Results from cell assays suggested that the observed growth promotion and wound healing effects of aloe substances in vitro may be analogous to what has been observed in vivo during healing of wounds and burns.     

Low activity of antioxidant enzymes in tumor cells as a factor of oxygen dependence of antitumor cytotoxicity of macrophages

The author studied dependence of cytotoxicity of peritoneal macrophages of Wistar rats on the production of active oxygen form (AOF) and activity of antioxidative enzymes in target cells. Low activity of glutathione-peroxidase and glutathione-reductase in K562 cells could be the cause of macrophage cytotoxicity realization along AOF-dependent way.     

Evaluation of antitumor properties of novel saframycin analogs in vitro and in vivo

Novel analogs of (-)-saframycin A are described. The analogs are shown to be potent inhibitors of the in vitro growth of several tumor cells in a broad panel and promising as leads for further optimization. The first in vivo studies in a solid tumor model (HCT-116) reveal potent antitumor activity with associated toxicity of daily administration.     

Tumouricidal activity of gilthead seabream (Sparus aurata L.) natural cytotoxic cells: the role played in vitro and in vivo by retinol acetate

The natural cytotoxic activity of gilthead seabream head-kidney leucocytes was evaluated after in vitro incubation with retinol acetate as vitamin A source, and in samples taken from specimens receiving an intraperitoneal injection or a diet supplemented with this vitamin. Isolated leucocytes were incubated with 0 to 10(-10)m all-trans-retinol acetate-supplemented culture medium for 0, 6 or 24h and assayed for their tumouricidal activity which was found to increase for all the assayed concentrations and incubation times. Seabream specimens were intraperitoneally injected with 0 (control), 1.75 or 5.25 micro g retinol acetate 100 g(-1) biomass and sampled 1, 3 or 5 days post-injection. Leucocyte natural cytotoxic activity increased in a dose-dependent manner 1 and 3 days post-injection. When fish were fed a commercial diet supplemented with 0 (control), 50, 150 or 300 mg retinol acetate kg(-1) diet for 1, 2, 4 or 6 weeks, only fish which had been fed the highest supplement for 2 weeks showed any increase in head-kidney leucocyte cytotoxic activity. Serum was isolated and analysed for all-trans-retinol concentration by reverse-phase high-pressure-liquid-chromatography. The normal level was about 0.4 micro g ml(-1) serum, while treatment for 1 to 4 weeks with vitamin A increased this level.In conclusion, retinol acetate increases gilthead seabream head-kidney leucocyte cytotoxic activity both in vitro and in vivo.