V. Oommen

From chemoprevention to chemotherapy: common targets and common goals.

Three decades of research have revealed that cancer is easier to prevent than to treat and that consumption of certain fruits and vegetables can reduce the risk of cancer. Whereas chemotherapy is designed to destroy cancer after it appears, chemoprevention involves the abrogation or delay in the onset of cancer. Regardless of whether a chemopreventive or chemotherapeutic approach is taken, cancer is a multifactorial disease that requires modulation of multiple pathways and multiple targets. Various molecular targets of chemoprevention are also relevant to the therapy of cancer. These targets include the activation of apoptosis; suppression of growth factor expression or signalling; downregulation of antiapoptotic proteins; suppression of phosphatidylinositol-3′-kinase/Akt, NF-κB, Janus kinase-signal transducer and activator of transcription and activator protein-1 signalling pathways; and downregulation of angiogenesis through inhibition of vascular endothelial growth factor expression, cyclooxygenase-2, matrix metalloproteinase-9, urokinase-type plasminogen activator, adhesion molecules and cyclin D1. Pharmacologically safe phytochemicals that have been identified from plants or their variant forms can modulate these molecular targets. These phytochemicals include genistein, resveratrol, dially sulfide, S-ally cysteine, allicin, lycopene, capsaicin, curcumin, 6-gingerol, ellagic acid, ursolic acid, betulinic acid, flavopiridol, silymarin, anethol, catechins and eugenol. Recent work has shown that these phytochemicals also can reverse chemoresistance and radioresistance. Because of their pharmacological safety, these agents can be used alone to prevent cancer and in combination with chemotherapy to treat cancer.

A molecular phylogeny of ichthyophiid caecilians (Amphibia: Gymnophiona: Ichthyophiidae): out of India or out of South East Asia?

Recent molecular phylogenetic studies indicate that the rafting Indian plate harboured several isolated vertebrate lineages between ca. 130 and 56 Myr ago that dispersed and diversified ‘out of Indi’ following accretion with Eurasia. A single family of the amphibian order Gymnophiona, the Ichthyophiidae, presently occurs on the Indian plate and across much of South East Asia. Ichthyophiid phylogeny is investigated in order to test competing out of India and out of South East Asia hypotheses for their distribution. Partial sequences of mitochondrial 12S and 16S rRNA and cytochrome b genes for 20 ichthyophiids and proximate outgroups were assembled. Parsimony, maximum–likelihood and distance analyses all recover optimum trees in which uraeotyphlids plus Ichthyophis cf. malabarensis are the sister taxa to all other Ichthyophis, among which the South East Asian taxa are monophyletic. Tree topology and branch lengths indicate that the Indian lineages are more basal and older, and thus are more consistent with the hypothesis that ichthyophiids dispersed from the Indian subcontinent into South East Asia. The estimated relationships also support monophyly of Sri Lankan Ichthyophis, and non–monophyly of striped and unstriped Ichthyophis species groups. Mitochondrial DNA sequences provide evidence that should assist current problematic areas of caecilian taxonomy.

Phylogenetic relationships of Indian caecilians (Amphibia: Gymnophiona) inferred from mitochondrial rRNA gene sequences.

India has a diverse caecilian fauna, including representatives of three of the six currently recognized families, the Caeciliidae, Ichthyophiidae, the endemic Uraeotyphlidae, but previous molecular phylogenetic studies of caecilians have not included sequences for any Indian caecilians. Partial 12S and 16S mitochondrial gene sequences were obtained for a single representative of each of the caecilian families found in India and aligned against previously reported sequences for 13 caecilian species. The resulting alignment (16 taxa, 1200 sites, of which 288 cannot be aligned unambiguously) was analyzed using parsimony, maximum-likelihood, and distance methods. As judged by bootstrap proportions, decay indices, and leaf stabilities, well-supported relationships of the Indian caecilians are recovered from the alignment. The data (1) corroborate the hypothesis, based on morphology, that the Uraeotyphlidae and Ichthyophiidae are sister taxa, (2) recover a monophyletic Ichthyophiidae, including Indian and South East Asian representatives, and (3) place the Indian caeciliid Gegeneophis ramaswamii as the sister group of the caeciliid caecilians of the Seychelles. Rough estimates of divergence times suggest an origin of the Uraeotyphlidae and Ichthyophiidae while India was isolated from Laurasia and Africa and are most consistent with an Indian origin of these families and subsequent dispersal of ichthyophiids into South East Asia.

Isodeoxyelephantopin, a Novel Sesquiterpene Lactone, Potentiates Apoptosis, Inhibits Invasion, and Abolishes Osteoclastogenesis through Suppression of Nuclear Factor-κB (NF-κB) Activation and NF-κB-Regulated Gene Expression

Purpose: Deoxyelephantopin (ESD) and isodeoxyelephantopin (ESI) are two sesquiterpene lactones derived from the medicinal plant Elephantopus scaber Linn. (Asteraceae). Although they are used for the treatment of a wide variety of proinflammatory diseases, very little is known about their mechanism of action. Because most genes that control inflammation are regulated by activation of the transcription factor nuclear factor-κB (NF-κB), we postulated that ESD and ESI mediate their activities through modulation of the NF-κB activation pathway. Experimental Design: We investigated the effect of ESI and ESD on NF-κB activation by electrophoretic mobility shift assay and NF-κB-regulated gene expression by Western blot analysis. Results: We found that ESI suppressed NF-κB activation induced by a wide variety of inflammatory agents, including tumor necrosis factor (TNF), interleukin-1β, phorbol 12-myristate 13-acetate, and lipopolysaccharide. The suppression was not cell type specific, and both inducible and constitutive NF-κB activation was blocked. ESI did not interfere with the binding of NF-κB to DNA but rather inhibited IκBα kinase, IκBα phosphorylation, IκBα degradation, p65 phosphorylation, and subsequent p65 nuclear translocation. ESI also suppressed the expression of TNF-induced NF-κB-regulated, proliferative, antiapoptotic, and metastatic gene products. These effects correlated with enhancement of apoptosis induced by TNF and suppression of TNF-induced invasion and receptor activator of NF-κB ligand-induced osteoclastogenesis. Conclusion: Our results indicate that ESI inhibits NF-κB activation and NF-κB-regulated gene expression, which may explain the ability of ESI to enhance apoptosis and inhibit invasion and osteoclastogenesis.

Thyroid hormones regulate lipid peroxidation and antioxidant enzyme activities in Anabas testudineus (Bloch).

The role of thyroid hormones in metabolic pathways are well known. However, their involvement in lipid peroxidation and antioxidant enzyme activities is not known. In this study, the in vivo injection of 6-propylthiouracil (6-PTU) did not alter the concentration of malondialdehyde (MDA) and conjugated dienes in liver. The administration of triiodothyronine (T3) or diiodothyronine (T2) increased the peroxidation rate in hypothyroid fish. However, in normal fish, only a high dose of T2 caused increased malondialdehyde (MDA) production, rather than T3. SOD activity was higher in T2-treated groups in both experiments. Glutathione peroxidase (GPx) activity was also high in hypothyroid fish treated with T2. In normal specimens, injections of T3 and T2 had no effect on GPx activity. Glutathione reductase (GR) activity was not altered by hypothyroidism while T3 (1 microg) and T2 (1 microg) increased it. Glutathione content was low in 6-PTU treated fish and high in both T3- and T2-treated groups. Thus it can be concluded that not only T3 but also T2, formed by sequential monodeiodination of T4, is also effective in influencing lipid peroxidation and antioxidant enzyme activities in Anabas. Furthermore, hypothyroidism as well as hyperthyroidism affects lipid peroxidation in this teleost.

Quantitative surveying of endogeic limbless vertebrates—a case study of Gegeneophis ramaswamii (Amphibia: Gymnophiona: Caeciliidae) in southern India

Many subterranean, limbless reptiles and amphibians are predators of invertebrate soil ecosystem engineers. The potential importance of these predators in soil ecology partly rests on whether they occur in high densities, but their abundance has rarely been measured, and there are no standard methods. The mostly tropical and fossorial caecilians (Amphibia: Gymnophiona) are often considered rare, but there are very few quantitative data, and some species, including Gegeneophis ramaswamii ,h ave been reported as abundant in some situations. Using simple and repeatable survey methods with randomised 1 m 2 quadrats, surveys of G. ramaswamii were conducted at five localities in southern India. Densities of 0–1.87 m −2 per survey were measured, with means of 0.51 and 0.63 m −2 at the beginning and middle of monsoon, respectively. These densities were far greater than for sympatric caecilians (ichthyophiids; uraeotyphlids) and fossorial snakes (typhlopids; colubrids). While ecological data remain very scant, establishing quantitative methods to assess the abundance of endogeic limbless vertebrates is an important step toward greater understanding of subterranean predator–prey relations, and of monitoring populations of these poorly known organisms.

Skeletal development of the direct-developing caecilian Gegeneophis ramaswamii (Amphibia: Gymnophiona: Caeciliidae)

A few previous studies of skeletal and especially skull development in Gymnophiona often provided contradictory results. We studied the development of the skull and vertebral column of Gegeneophis ramaswamii, a direct-developing Indian caeciliid, based on 13 specimens. The chondrocranium forms at (Brauer in Zool Jahrb Anat 12:477-508,1899) stage 38. First dermal and perichondral ossifications occur at stage 40. The first dermal bones to form are the mentomeckelian, dentary, angular, vomer, and premaxillary. These are followed by the coronoid, palatine, pterygoid, maxillary, and the skull-roofing bones. The last occurring dermal ossifications are the parasphenoid and the squamosal. We present evidence for the occurrence of a lacrimal bone. No ectopterygoid, basioccipital, supraoccipital, pleurosphenoid, postorbital, or supratemporal elements were found. We assess the homology of the bones constituting the caecilian skull and discuss the above-mentioned terminologies. The phylogenetic implications of our findings are briefly discussed and we conclude that the evidence from developmental morphology is at present consistent with a monophyletic Lissamphibia of temnospondyl origin.

Rapid action of glucocorticoids on branchial ATPase activity in Oreochromis mossambicus: an in vivo and in vitro study.

The rapid action of cortisol and corticosterone on branchial Na(+)-K(+) ATPase, Ca(2+) ATPase activity and Na(+), K(+) and Ca(2+) ion contents was studied both in vivo and in vitro employing transcription inhibitor actinomycin D in Oreochromis mossambicus. Cortisol and corticosterone administration had significantly increased the activity of branchial Na(+)-K(+) ATPase and Ca(2+) ATPase in vivo after 30 min of injection, and the trend continued for 60 and 120 min for cortisol. The ionic contents were also significantly increased after 30 min in vivo. Na(+)-K(+) ATPase activity was significantly increased 5 min after hormone application in the in vitro system. Actinomycin D did not inhibit the effect of glucocorticoids on ATPase activity both in vivo and in vitro. It is concluded from the present study that cortisol and corticosterone produced a rapid stimulatory effect on branchial ATPase activity and ions in O. mossambicus both in vivo and in vitro. This effect could be due to a non-genomic action of these hormones since the enzyme activity was insensitive to actinomycin D.

Rapid action of cortisol and testosterone on lipogenic enzymes in a fresh water fish Oreochromis mossambicus: short-term in vivo and in vitro study

Rapid action of steroid hormones on lipid metabolism is not reported so far in any vertebrate. The present study was intended to evaluate the quick actions of cortisol and testosterone on enzymes, namely malic enzyme (ME), glucose-6-phosphate dehydrogenase (G6PDH), and isocitrate dehydrogenase (ICDH) in Oreochromis mossambicus. Cortisol and testosterone produced rapid and opposite effects on the lipogenic enzymes studied. Cortisol significantly decreased the activities of ME, G6PDH, as early as 5 min and ICDH as early as 10 min in vitro (10(-6) M), and 30 min in vivo (0.1 microg/g body wt.) whereas the same doses of testosterone significantly stimulated the activity of all enzymes as early as 5 min in vitro and 30 min in vivo. Actinomycin D treatment did not interfere with the inhibiting effect of cortisol on enzyme activities when measured at 10 min in the in vitro system. The transcriptional inhibitor appeared to partially block the effect of cortisol in vivo. The stimulatory effect of testosterone was insensitive to the action of actinomycin D both in vivo and in vitro. These effects appear to be brought about independently of new protein synthesis because the rapid responses occurred within a latent period of 5-30 min and were insensitive to the action of actinomycin D, suggesting a non-genomic action.

Long-term feeding of dietary oils alters lipid metabolism, lipid peroxidation, and antioxidant enzyme activities in a teleost (Anabas testudineus bloch).

Anabas testudineus (climbing perch), average body weight 21±1 g, were maintained in culture tanks and fed a 35% protein feed plus an additional supplementation of three dietary oils (20% each of coconut oil, palm oil, or cod liver oil). Body weight gain was similar among all groups. However, several hepatic lipogenic enzymes such as malic enzyme (ME), NADP-isocitrate dehydrogenase (ICDH), glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH) and β-hydroxy-β-methyl glutaryl CoA reductase (HMG CoA reductase) were assayed, and they responded differently. Hepatic ME and G6PDH activities showed a significant decrease in the coconut oil and palm oil groups, but there was no significant change in ICDH activity. The 6PGDH activities were reduced, whereas HMG CoA reductase activity was increased in the palm oil-treated group. Cholesterol synthesis in the liver and muscle increased in the palm oil-treated group, but liver phospholipids did not show any significant change in fish supplemented with oils rich in saturated fatty acids. Triacyl-glycerol and free fatty acid concentrations were high in the coconut oil- and palm oil-supplemented groups. Lipid peroxidation products such as thiobarbituric acid-reactive substances and conjugated dienes decreased in the same two groups. Antioxidant potential was high in all groups as evidenced by increased activity of superoxide dismutase, glutathione peroxidase, and glutathione content. The results of this study indicate that in fish, dietary lipids depress hepatic lipogenic activity as well as lipid peroxidation products by maintaining high levels of antioxidant enzymes.