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Trabajo presentado al: 73rd Annual Meeting of the European Federation of Animal Science (EAAP), Oporto (Portugal), 5-9 de septiembre. 2022.

Trabajo presentado al: 73rd Annual Meeting of the European Federation of Animal Science (EAAP). Oporto (Portugal), 5-9 de septiembre.

Trabajo presentado al: 73rd Annual Meeting of the European Federation of Animal Science (EAAP), Oporto (Portugal), 5-9 de septiembre.

Currently, upon understanding the metabolomics of carotenoids, it is important to address the key role of carotenoid derived products. In this regard, aim of the study was to elucidate and explore the role of lycopene (LYC) oxidative products generated through autoxidation (AOL) or chemical (KMnO4) oxidation (COL) against proliferation of selected cancer cells. Preliminary, we investigated the effect of LYC on cell viability of various cancer cell lines (PC-3, MCF-7, A431, HepG(2), HeLa and A549). Based on the results of LYC treatment on cell cytotoxicity levels, MCF-7, PC-3 and HeLa cell lines were further tested with AOL and COL products. The decreased cell viability with depleted GSH and increased MDA levels were observed when treated with COL products than control, LYC and AOL In addition, COL products increased ROS levels and percent apoptosis. The typical morphological changes and nuclear condensations showed that COL products have anti-proliferation and apoptosis inducing activity. Based on results, we hypothesized that ROS generation by LYC oxidation products may be one of intermediate step involved in apoptosis. The redox status and therapeutic approach of COL products in modulating ROS and induction of apoptosis in cancer cells were reported for the first time, to our knowledge. To conclude, COL products involves in cancer growth inhibition efficiently than intact LYC and AOL. Hence, there is a great potential for synthesizing or producing such carotenoid oxidation products to augment cancer complication. (C) 2016 Elsevier Ltd. All rights reserved.

Search for novel anticancer lead molecules continues to be a major focus of cancer research due to the limitations of existing drugs such as lack of tumor selectivity, narrow therapeutic index and multidrug resistance of cancer types. Natural molecules often possess better pharmacokinetic traits compared to synthetic molecules as they continually evolve by natural selection process to interact with biological macromolecules. Microbial metabolites constitute nearly half of the pharmaceuticals in market today. Endophytic fungi, owing to its rich chemical diversity, are viewed as attractive sources of novel bioactive compounds. In the present study, we report the purification and characterization of a novel steroidal saponin, cholestanol glucoside (CG) from Saraca asoca endophytic fungus Lasiodiplodia theobromae. The compound was assessed for its cytotoxic potentialities in six human cancer cell lines, A549, PC3, HepG2, U251, MCF7 and OVCAR3. CG exhibited significant cytotoxicities towards A549, PC3 and HepG2 among which A549 cells were most vulnerable to CG treatment. However, CG treatment exhibited negligible cytotoxicity in non malignant human lung fibroblast cell line (WI-38). Induction of cell death by CG treatment in A549 cells was further investigated. CG induced the generation of reactive oxygen species (ROS) and mitochondrial membrane permeability loss followed by apoptotic cell death. Mitochondrial membrane depolarization and apoptotic cell death in CG treated A549 cells were completely blocked in presence of an antioxidant, N-acetyl cysteine (NAC). Hence it could be concluded that CG initiates apoptosis in cancer cells by augmenting the basal oxidative stress and that the generation of intracellular ROS is crucial for the induction of apoptosis.

Lycopene (LYC) has been correlated with the reduction of certain cancers and chronic diseases. However, the existence and biofunctionality of degraded, oxidized, and biotransformed LYC products in vivo have not been revealed. Therefore, this study aimed to screen and elucidate the potential bioactive lycopene-derived products in breast-cancer and noncancerous cells. LYC-oxidation or -cleavage products were generated using KMnO4. These oxidation products were separated as fractions I-III by silica column chromatography using gradient solvent systems. Further, LC-MS/MS (ESI)(+) was used to elucidate their possible fragmentation patterns and structures. Fraction II showed higher cytotoxicity (IC50 value of 64.5 mu M), cellular uptake, and apoptosis-inducing activity in MCF-7 cells. This fraction consists of major peak m/z 323, identified as apo-8,6'-carotendial. The cytotoxicity-inducing activity may be due to partial ROS generation with mitochondrial dysfunction. Further, the role of apo-8,6'-carotendial in the induction of apoptosis is demonstrated for the first time. These results illustrated that LYC-oxidation derivatives or metabolites are involved in growth inhibition of cancer cells. Exploration of specific oxidized-carotenoid products will give further insight into the field of nutritional biochemistry.

Seventeen endophytic fungi were isolated from various tissues of Cassia fistula and the ethyl acetate extracts obtained from 21-day cultures of all the endophytic fungal isolates were initially screened for their cytotoxicity against HeLa (human cervical carcinoma) cells using MTT assay. Of these, Penicillium sclerotiorum extract (PSE), significantly affected the viability of HeLa cells in a dose-dependent manner. The extract of P. Sclerotiorum was further analyzed by GC–MS, which showed three compounds, hexadecanoic acid, oleic acid and benzoic acid to be the major active principles in the extracts.The extract was further tested for invitro cytotoxicity against five cancer cell lines. Of the cell lines tested, HeLa cells showed maximum sensitivity followed by A549, while A431 and U251 were moderately sensitive and MCF-7 was insensitive to the treatment. In addition, normal human embryonic kidney cells, HEK293 remained insensitive to the treatment. Furthermore, the mechanism of cytotoxic activity exhibited by PSE was investigated by evaluating cell cycle progression and apoptotic induction in HeLa cells. Cell cycle analysis revealed that the PSE arrested cells at S and G2/M phase of the cell cycle in a dose-dependent manner. Annexin V- Propidium iodide double staining showed that, the extract potentiates apoptosis rather than necrosis in cells. This was supported by the down regulation in the proapoptotic protein BCL2 and up regulation of BAX (BCL2 Associated X), tumor suppressor protein, p53 and Apaf-1 [Apoptotic Peptidase Activating Factor 1]. Loss of mitochondrial membrane potential and a distinct DNA fragmentation pattern observed following the treatment, suggest that the PSE treatment leads to activation of mitochondrial pathway of apoptosis. Further, the extract also exhibited both antioxidant and anti-angiogenic properties. These results indicate that endophytic fungi isolated from medicinal plants may serve as potential sources of the anti-cancerous compounds.

Endophytic fungi isolated from Catharanthus roseus were screened for the production of vincristine and vinblastine. Twenty-two endophytic fungi isolated from various tissues of C. roseus were characterized taxonomically by sequence analysis of the internal transcribed spacer (ITS) region of rDNA and grouped into 10 genera: Alternaria, Aspergillus, Chaetomium, Colletotrichum, Dothideomycetes, Eutypella, Eutypa, Flavodon, Fusarium and Talaromyces. The antiproliferative activity of these fungi was assayed in HeLa cells using the MTT assay. The fungal isolates Eutypella sp-CrP14, obtained from stem tissues, and Talaromyces radicus-CrP20, obtained from leaf tissues, showed the strongest antiproliferative activity, with IC50 values of 13.5 mu g/ml and 20 mu g/ml, respectively. All 22 endophytic fungi were screened for the presence of the gene encoding tryptophan decarboxylase (TDC), the key enzyme in the terpenoid indole alkaloid biosynthetic pathway, though this gene could only be amplified from T. radicus-CrP20 (NCBI GenBank accession number KC920846). The production of vincristine and vinblastine by T. radicus-CrP20 was confirmed and optimized in nine different liquid media. Good yields of vincristine (670 mu g/l) in modified M2 medium and of vinblastine (70 mu g/l) in potato dextrose broth medium were obtained. The cytotoxic activity of partially purified fungal vincristine was evaluated in different human cancer cell lines, with HeLa cells showing maximum susceptibility. The apoptosis-inducing activity of vincristine derived from this fungus was established through cell cycle analysis, loss of mitochondrial membrane potential and DNA fragmentation patterns.

The article deals with the sociology of labour. The author accepts the psychological criteria of differentiation of social groups and depicts the evolution of society and of the spirit of accumulation in relation to the changes that have occurred in the psychology of man the worker, throwing into relief the main stages of this development. The importance of this last stage - that of spontaneous and pleasurable work, of which the United States represents the most advanced type among the Caucasian peoples - is such as to induce the author to suggest once again the need for an economic science wider and more comprehensive than “bourgeois” economics, which he calls Integral Economics: this should be based on a more general theoretical outline covering the whole of the historical development of human psychology and institutions. JEL: J01, J20, Z13, PSL Quarterly Review, V. 1, N. 4 (1948)

The article discusses national income as it presents itself in pathological periods, and as seen more especially in the light of post-war experiences. From the statistical-theoretical angle where practical interests and economic inquiries converge and bring into focus the real problems of income, consumption, savings, investments, and their mutual relations, we find that even the most subtle premises of methodology need to be studied when framing plans for solving the problems of present day reconstruction and taxation. English-speaking students of such questions will note with interest the references the author makes to Pigou’s recent work on “Income” and the restatement of the opinions the author himself had occasion to sustain before the Royal Statistical Society in London in 1939. JEL: E01, PSL Quarterly Review, V. 1, N. 2 (1947)

The article shows that in the efforts to re-establish normal international relations on the basis of multilateral trade, confusion has often arisen concerning the distinction between those which are necessary effects or conditions, and those which are sufficient conditions, and that the disadvantages of bilateral trade have also been exaggerated. The inconvertibility of national currencies was not the cause but the effect of the system of bilateral trade, and the revival of the system of multilateral trade would not suffice to re-establish convertibility. The effects of agreements between groups of States for the purpose of assuring the periodical clearing of active and passive balances are also very limited. The most objectionable feature of bilateral treaties is that they deviate commercial currents from their most economically expedient directions, but they do not necessarily alter substantially the final distribution of the goods. Nor is it true that the system of bilateral trade necessarily determines the divergence, noted in some countries, between direct and indirect exchanges, which may be due rather to the fact that the exchanges are fixed artificially at a level other than the equilibrium rate. JEL: F10, F13, F31, PSL Quarterly Review, V. 2, N. 8 (1949)

The article compares the theory of saving, traditional in Europe, with the anti-saving theory formulated by Ford, which advocates spending all earning, thus stimulating production, raising the standard of living, and giving confidence in the future. The author shows that the two theories correspond to the different economic conditions prevailing in Europe and the United States. The Americans are not wealthy and optimistic because they spend all they earn: rather they spend all they earn and are optimistic because they are wealthy. The fundamental cause for the dazzling economic prosperity of the United States is to be found instead in the free contribution of wealth represented by the masses of emigrants, in the great majority Europeans, thanks to whom America has been saved the expense of bringing up a large part of her population. JEL: E21, PSL Quarterly Review, V. 1, N. 6 (1948)

The brief note makes a contribution to the methodology to be used in calculating the national income. The author shows that there is no alternative choice between the several price index numbers with which to value the commodities consumed by the producer himself. The difficulties involved in the valuation are indeed solved by making a simple analysis of structure of the price general index number. The solution therefore comes spontaneously, and this index number is proposed as the only possible one to be used in making the calculation. JEL: E01, E21, E31, PSL Quarterly Review, V. 2, N. 9 (1949)

The work is the second of two articles, the first of which was published under the title “The Mask of Government”, in which the author presents the results of his new inquires into the theory and calculation of the national income. The present work looks at the problem of finding a unit of measure for the purchasing power of money which will remain stable, and will therefore give precise meaning to its variations in successive periods. The author’s conclusion is that it is not possible to find such a satisfactory minimum common denominator of the purchasing power of several goods in different countries because of the disparities in the remuneration of labour which arise form the fact that labour cannot move freely in space. JEL: E01, E24, PSL Quarterly Review, V. 5, N. 20 (1952)

The article is the first of two enquires by the author into the theory and calculation of national income. Here, he deals with certain problems concerning the relations between services or goods of State or Public Corporations and those of private persons. JEL: E01, PSL Quarterly Review, V. 4, N. 19 (1951)

The note takes up a position definitely critical of an indiscriminate application of Truman’s Point IV and, in particular, to its extension to tropical and equatorial lands. This controversial attitude is based on an outright appreciation of the danger which might arise for the supremacy of the Western peoples and Western civilisation. JEL: O11, O19, O20, PSL Quarterly Review, V. 4, N. 17 (1951)

A comment on Dr. Gioacchino d’Ippolito’s article on the problem of the concordance of direct exchange rates and cross-rates under a system of bilateral trade. JEL: F10, F31, PSL Quarterly Review, V. 3, N. 12 (1950)

Moneta e Credito, V. 1, N. 2 (1948)

Moneta e Credito, V. 1, N. 4 (1948)

Objectives: Endophytic strains of many plants-particularly medicinal plants-produce a plethora of secondary metabolites, many of which are of immense pharmaceutical importance e.g. anticancer properties. Datura metel L., an important medicinal plant in use as a topical application to remove tumors apart from its other medicinal uses, has not been extensively explored for the potential of its endophytes for their pharmacological significance. The present study was thus initiated to investigate the anticancer effects of the organic extract of an endophytic fungus isolated from this plant. Methods: We report the anticancer effects of an endophytic Fusarium solani fungal strain isolated from Datura metel L. as evaluated by its culture extract on five human cancer cell lines (HepG2, HeLa, MCF-7, OVCAR-3 and PC-3). The ethyl acetate (EtOAc) extract of three week grown fungal culture was tested for its cytotoxic activity on different cancer cell lines by MTT ([3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The apoptosis-inducing activity and its effect on mitochondrial membrane potential were measured by flow cytometry using JC-1 dye. Nuclear DNA condensation was assessed by fluorescence microscopy using Hoechst 33342 stain. DNA fragmentation was visualized by gel electrophoresis. Results: The ethyl acetate (EtOAc) fungal culture extract showed cytotoxic activity against all the tested human cancer cell lines, in particular against cervical cancer cells HeLa. Further, loss of mitochondrial membrane potential, DNA fragmentation and nuclear chromatin condensation strongly support the ability of organic extract to induce the cancer cell apoptosis though the mitochondrial pathway. Conclusion: The results demonstrate that the Fusarium solani organic extract harbor potential anticancer lead compound(s) which inhibited the proliferation of various cancer cells by inducing cell apoptosis.

Background: Cancer is one of the leading causes of morbidity and mortality globally. Cancer-associated thrombosis is well established in clinical settings, and thrombin has been found to induce angiogenesis at cancer sites. This establishes a link between cardiovascular diseases and cancer, where cancer and thrombin have been intricately associated. Various selective estrogen receptor modulators (SERMs) have been reported to exhibit anticancer activity. Therefore, we investigated estradiol-17β and SERMs dl-ormeloxifene (centchroman), raloxifene and levormeloxifene (l-centchroman) for their anticancer effects and their effect on thrombin activity. Methods: Anticancer activity was assessed against PC-3 cell line by flow cytometry following treatment with estradiol-17β and SERMs at 10 nM-1 mM concentrations. The cells were stained with propidium iodide and the percentage of cells in the sub-G0/G1 region was considered apoptotic. Thrombin inhibitory effect was evaluated by thrombin inhibition assay in vitro following incubation with 100 nM-3 mM concentrations of estradiol-17β or various SERMs. Further, the effect of estradiol-17β and SERMs on endogenous thrombin generation potential (ETP) was assessed by thrombin generation assay on rat plasma in vitro. Results: These compounds exhibited >90% cell death in PC-3 cell lines at 1 mM concentration except estradiol-17β. Neither estradiol-17β, dl-ormeloxifene and levormeloxifene showed any thrombin inhibitory or enhancing activity in thrombin inhibition assay, nor did they show any effect on ETP on rat plasma in vitro. However, raloxifene inhibited thrombin activity in a concentration-dependent manner. Raloxifene decreased ETP of the plasma at 3 and 1 mM,which is equivalent to that of 30-100 U/mL of heparin. Interestingly, raloxifene increased thrombin generation at lower concentrations and it inhibited thrombin generation at higher concentrations. Conclusions: These observations suggest that dl-ormeloxifene, estradiol-17β and levormeloxifene do not possess thrombin inhibitory activity. Raloxifene possesses thrombin modulatory effect in addition to its anticancer activity, and this observation may help us in understanding the thromboembolic complications associated with raloxifene.

The vast biodiversity of nature provides bioactive compounds that may be useful in the fight against chronic diseases. This study was designed to investigate the protective effects of the ethanol extract of Spirulina laxissima West (Pseudanabaenaceae) (EESL) against carbon tetrachloride (CCl(4)) induced hepatotoxicities in rats. Male albino rats of Sprague-Dawley strain were treated orally with the ethanol extract of S. laxissima (50, 100 mg kg(-1) body wt.) 1 h before each CCl(4) administration. The ethanol extract of S. laxissima showed the maximum antioxidant property in vitro. There were statistically significant losses in the activities of antioxidant enzymes and an increase in TBARS and liver function marker enzymes in the serum of the CCl(4)-treated group compared with the control group. However, all the tested groups were able to counteract these effects. The antioxidant activity of the extracts might be attributable to its proton-donating ability, as evidenced by DPPH. In the present study, the decline in the level of antioxidant observed in CCl(4)-treated rats is a clear manifestation of excessive formation of radicals and activation of the lipid peroxidation system resulting in tissue damage. The significant increases in the concentration of antioxidant enzymes in tissues of animals treated with CCl(4) + EESL indicate the antioxidant effect of EESL. This study suggests that EESL can protect the liver against CCl(4)-induced oxidative damage in rats, and the hepatoprotective effect might be correlated with its antioxidant and radical-scavenging effects.

Paracetamol caused liver damage as evident by significant increase in the activities of aspartate and alanine transferases. There were general statistically significant losses in the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione transferase and an increase in thiobarbituric acid reactive substances in the liver of paracetamol treated group compared with the control group. However, treatment with ethanol extract of A. flos-aquae (EEAFA) was able to counteract these effects. Protection offered by silymarin (standard reference drug) seemed relatively greater. The results suggest that EEAFA can act as hepatoprotective agent against paracetamol induced toxicity as an antioxidant.

Oxidative stress due to abnormal production of reactive oxygen molecules (ROM) is believed to be involved in the etiology of toxicities of many xenobiotics. Evidence suggested that ROM is involved in the nephrotoxicity of a widely used synthetic anticancer drug cisplatin. The nephroprotective effects of ethanol extract of Aulosira fertilisima Ghose (EEA) was evaluated using cisplatin (5 mg/kg(-1) i.p.)-induced renal damage in rats. EEA showed higher significant effect on DPPH radical scavenging activity as compared with methanol extract of A. fertilisima (MEA) and water extract of A. fertilisima (WEA). Thus, EEA was selected for further in vivo studies. The serum urea and creatinine levels in the cisplatin alone-treated group were significantly elevated with respect to normal group of animals. The levels were reduced in the EEA (100 mg/kg, p.o) plus cisplatin-treated groups. Renal oxidative stress was determined by renal TBARS, CD and reduced glutathione levels, and by enzymatic activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione transferase (GST). A single dose of cisplatin-produced marked renal oxidative and nitrosative stress and significantly deranged renal functions. Chronic EEA treatment significantly and dose-dependently restored renal functions, reduced lipid peroxidation, and enhanced reduced glutathione levels, superoxide dismutase, and catalase activities. The results of the study indicated that A. fertilisima significantly and dose-dependently protected the nephrotoxicity induced by cisplatin. This protection is mediated either by preventing the cisplatin-induced decline of renal antioxidant defense system or by their direct free radical scavenging activity.