Organoruthenium metallocycle induced mutation in gld-1 tumor suppression gene in JK1466 strain and appreciable lifespan expansion.

Four Ru(II) complexes (A2-A5) were synthesized from the reaction of coumarin Schiff base ligands (7da2-tsc, 7da3-mtsc, 7da4-etsc and 7da5-ptsc) with [RuHCl(CO)(PPh3)3]. The compounds were characterized by FT-IR, UV-Vis, 1H, 13C and 31P NMR, mass spectrometry and crystallographic analysis. Calf Thymus DNA (CT-DNA) binding studies revealed the intercalative mode of binding of the complexes with DNA. The results of Bovine serum albumin (BSA) binding studies established the interaction between BSA followed static quenching mechanism. The cytotoxic effects of the complexes and the ligands were evaluated against breast (MCF-7 and MDA-MB-231) and lung carcinoma cell lines (A549 and NCI-H460) using MTT assay. Complex A4 demonstrated potent cytotoxic effects on both breast and lung cancer cells. Furthermore, morphological observations and FACS analysis showed the decrease in cell density by complex A4 by induced morphological changes and apoptotic body formation and cell death in both breast and lung cancer cells. Moreover, the invertebrate model Caenorhabditis elegans was employed to assess the in vivo anticancer activity of compound A4. The findings indicated that the treatment with A4 reduced tumor development and significantly extended organismal lifespan by 64 % in the tumoral strain JK1466 without adversely affecting essential physiological functions of the worm. Additionally, A4 demonstrated an upregulation of two crucial antioxidant defense genes. Overall, these results suggested that the compound A4 can be a potential candidate with novel chemotherapeutic applications.

Effect of ruthenium(II) complexes on MDA-MB-231 cells and lifespan/tumor growth in gld-1mutant, Daf-16 TF and stress productive genes: A perspective study.

Pincer type coumarin based N-substituted semicarbazone ligands HL1-4 and their corresponding ruthenium(II) complexes (1-4) were synthesized, analyzed and confirmed by various spectro analytical techniques. The molecular structure of the ligand HL3 and complex 3 was confirmed by single crystal X-ray diffraction analysis. The stoichiometry of complexes 1, 2 and 4 was confirmed by high resolution mass spectroscopy (HRMS). The binding affinity of the compounds with CT-DNA (Calf Thymus DNA) and BSA (Bovine Serum Albumin) was established by absorption and emission titration methods. The results of In vitro cytotoxicity showed the significant cytotoxic potential of the complexes against MDA-MB-231 cells (TNBC- Triple-negative breast cancer). Among the complexes, 1 and 4 have shown appreciable results. Further, antimigratory activity against the MDA-MB-231 cells was studied for the complexes 1 and 4. The percentage cell cycle arrest, apoptosis and necrosis were explored by flow cytometry. The in vivo anti-tumor activity of the complexes 1 and 4 using C. elegans as model organism was established by using the tumoral C. elegans strain JK1466 (gld-1(q485)), which bears a mutation in the gld-1 tumor suppressor gene. We have determined the effect of our complexes on tumor gonad reduction and found to be non toxic to the JK1466 worms and they have prolonged their mean lifespan with potential antioxidant ability by overcoming stress responses. Overall, our study reported herein demonstrated that the complexes 1 and 4 could be established as potential metallo-drugs substantiating further exploration.

An aggregation-induced emission-based ratiometric fluorescent chemosensor for Hg(II) and its application in Caenorhabditis elegans imaging.

A chromone-based ratiometric fluorescent probe L2 was developed for the selective detection of Hg(II) in a semi-aqueous solution based on aggregation-induced emission (AIE) and chelation-enhanced fluorescence (CHEF) effect. The probe L2 fluoresced significantly at 498 nm in its aggregated state, and when chelated with Hg(II), the soluble state fluoresced 1-fold higher. In addition, Job's plot reveals that the probe forms a 1:1 stoichiometry complex with Hg(II) with an association constant of 9.10 × 103M-1 estimated by the BH plot. The probe L2 detects Hg(II) down to 22.47 nM without interference from other interfering ions. The FTIR, ESI mass, and DFT-based computational studies investigated the binding mechanism of probe L2 with Hg(II). Taking advantage of its AIE characteristics, the probe L2 was successfully applied for bio-capability analysis in Caenorhabditis elegans (a nematode worm) imaging of Hg(II) in a living model.

Effect of gamma-ray irradiated reduced graphene oxide (rGO) on environmental health: An in-vitro and in-vivo studies.

The unique properties of reduced graphene oxide (rGO) have drawn the attention of scientists worldwide since the last decade and it is explored for a wide range of applications. However, the rapid expansion of rGO use in various products will eventually lead to environenal exposure and rises a safety concern on the environment and humal health risk. Moreover, the utilization of toxic chemicals for the reduction of graphene oxide (GO) into rGO is not environmentally friendly, warranting the exploration of non-toxic approaches. In the present work, rGO was synthesized using a different dose of gamma-ray irradiation and characterized. The in-vitro and in-vivo analysis indicated that the gamma-irradiated rGO induced toxicity depending on its degree of reduction and dosage. In the L929 cells, rGO-30 KGy significantly induced cytotoxicity even at low concentration (1 mg L-1) by inducing reactive oxygen species (ROS), lactate dehydrogenase (LDH) enzyme production, nuclear fragmentation and apoptosis. The change in morphology of the cells like membrane blebbing and cell rounding was also observed via FESEM. In the in-vivo model Caenorhabditis elegans, rGO-30 KGy significantly affected the functioning of primary and secondary targeted organs and also negatively influenced the nuclear accumulation of transcription factors (DAF-16/FOXO and SKN-1/Nrf2), neuronal health, and antioxidant defense mechanism of the nematodes. The real-time PCR analysis showed significant up-regulation (ced-3, ced-4, cep-1, egl-1, and hus-1) and down-regulation (ced-9) of the gene involved in germ-line and DNA damage-induced apoptosis. The detailed toxicity mechanism of gamma irradiated rGO has been elucidated. This work highlights the toxicity of rGO prepared by gamma-ray radiation and paves way for understating the toxicity mechanism.

Dual sensing of methionine and aspartic acid in aqueous medium by a quinoline-based fluorescent probe.

A quinoline-based Schiff base sensor, 6-methyl-2-oxo-1,2-dihydro-quinoline-3-carboxaldehyde-4(N)-phenylsemicarbazone (6MPS), has been developed for selective sensing of methionine and aspartic acid in aqueous medium through "on-off-on" type selective detection of copper ion. Fluorescence imaging of 6MPS, 6MPSC, 6MPSCN, 6MPSC-met, 6MPSCN-met, 6MPSC-asp and 6MPSCN-asp has been successfully demonstrated, in which the sensing probes 6MPSC-met, 6MPSCN-met, 6MPSC-asp and 6MPSCN-asp displayed bright green fluorescence in both in vitro and in vivo live cells.

Engineering the surface of graphene oxide with bovine serum albumin for improved biocompatibility in Caenorhabditis elegans.

Graphene oxide (GO) has been extensively studied for its potential biomedical applications. However, its potential risk associated with the interactions of GO in a biological system hampers its biomedical applications. Therefore, there is an urgent need to enhance the biocompatibility of GO. In the present study, we decorated the surface of GO with bovine serum albumin (GO-BSA) to mitigate the in vivo toxic properties of GO. An in vivo model Caenorhabditis elegans has been used to study the potential protective effect of BSA decoration in mitigating GO induced toxicity. The BSA decoration on the surface of GO prevents the acute and prolonged toxicity induced by GO in primary and secondary organs by maintaining normal intestinal permeability, defecation behavior, development, and reproduction. Notably, GO-BSA treatment at 0.5-100 mg L-1 does not affect the intracellular redox status and lifespan of C. elegans. Reporter gene expression analysis revealed that exposure to GO-BSA (100 mg L-1) did not significantly influence the nuclear accumulation and expression patterns of DAF-16/FOXO and SKN-1/Nrf2 transcription factors and their downstream target genes sod-3, hsp-16.2, ctl-1,2,3, gcs-1, and gst-4 when compared to exposure to pristine GO. Also, quantitative real-time PCR results showed that GO-BSA did not alter the expression of genes involved in regulating DNA damage checkpoints (cep-1, hus-1 and egl-1) and core signaling pathways of apoptosis (ced-4, ced-3 and ced-9), in contrast to GO treatment. All these findings will have an impact on the future development of safer nanomaterial formulations of graphene and graphene-based materials for environmental and biomedical applications.

Multiple infestations of gastrointestinal parasites - Probable cause for high mortality of Spot-billed Pelican (Pelecanus philippensis) at Kokrebellur Community Reserve, India.

We witnessed mortalities of Spot-billed Pelicans Pelecanus philippensis between December 2017 and May 2018 in Mandya and Mysuru districts of Karnataka, especially at Kokrebellur Community Reserve in Mandya district. The region has experienced severe drought in recent years with negligible water in all the water tanks. A total of 67 Spot-billed Pelicans died in five locations, of which 55 adult birds died at Kokrebellur. We collected four dead pelicans along with 97 fecal samples of live birds at Kokrebellur, water samples from nine water tanks around Kokrebellur, and six fish samples. We isolated the endoparasite eggs by following sedimentation and flotation technique, and counted the eggs from the water and fecal samples, and identified at the genus level using light microscope. We approximately counted the endoparasites by dissecting the fish and conducting a necropsy on dead pelicans. Endoparasite eggs were detected in seven of the nine water tanks. Each fish sample had at least 50-100 L3 stage worms of Contracaecum sp., and 880.0 ±â€¯459.3SD of Contracaecum sp., worms in the digestive tracts and 60.0 ±â€¯36.5SD worms of Echinostoma sp. in the intestine of the four dead pelicans. The endoparasite prevalence was 84.5% (N = 83) with a mean abundance of 368.2 ±â€¯561.5SD eggs/g in the fecal samples of live pelicans. Contracaecum sp., Echinostoma sp. and Opisthorchis viverrini were recorded in 51, 67 and nine fecal samples respectively. The high load of endoparasite eggs in the water tanks, an infestation of Contracaecum sp. in fishes and a heavy load of fully-grown worms of Contracaecum sp. and Echinostoma sp. in the adult pelicans are indicative of their high mortality in Kokrebellur Community Reserve. The coordinated program was initiated with the support of all stakeholders to control the endoparasites in water, fish, and pelicans.

Organoruthenium(II) complexes attenuate stress in Caenorhabditis elegans through regulating antioxidant machinery.

The 1:1 stoichiometric reactions of 3-methoxy salicylaldehyde-4(N)-substituted thiosemicarbazones (H2L1-4) with [RuCpCl(PPh3)2] was carried out in methanol. The obtained complexes (1-4) were characterized by analytical, IR, absorption and 1H NMR spectroscopic studies. The structures of ligand [H2-3MSal-etsc] (H2L3) and complex [RuCp(Msal-etsc) (PPh3)] (3), were characterized by single crystal X-ray diffraction studies. The interaction of the ruthenium(II) complexes (1-4) with calfthymus DNA (CT-DNA) has been explored by absorption and emission titration methods. Based on the observations, an intercalative binding mode of DNA has been proposed. The protein binding abilities of the new complexes were monitored by quenching the tryptophan and tyrosine residues of BSA, as model protein. From the studies, it was found that the new ruthenium metallacycles exhibited better affinity than their precursors. The free radical scavenging assay suggests that all complexes effectively scavenged the DPPH radicals as compared to that of standard control ascorbic acid and scavenging activities of complexes are in the order of 4 > 2 > 3 > 1. In addition, ruthenium(II) complexes (2-4) also exhibited an excellent in vivo antioxidant activity as it was able to increase the survival of worms exposed to lethal oxidative and thermal stresses possibly through reducing the intracellular ROS levels. It was interesting to note that complexes 2-4 failed to increase the lifespan of mev-1 mutant worms having shortened lifespan due to the over production of free radicals. This data confirmed that complexes 2-4 conferred stress resistance in C. elegans, but they also require an endogenous detoxification mechanism for doing so. The genetic and reporter gene expression analysis revealed that complexes 2-4 maintained the intracellular redox status and offered stress protection through transactivation of antioxidant defence machinery genes gst-4 and sod-3 which are directly regulated by SKN-1 and DAF-16 transcription factors, respectively. Altogether, our results suggested that complexes 2-4 might play a crucial role in stress modulation and they perhaps exert almost similar effects in higher models, which is an important issue to be validated in future.

East Indian sandalwood (Santalum album L.) oil confers neuroprotection and geroprotection in Caenorhabditis elegans via activating SKN-1/Nrf2 signaling pathway.

East Indian Sandalwood Oil (EISO) has diverse beneficial effects and has been used for thousands of years in traditional folk-medicine for treatment of different human ailments. However, there has been no in-depth scientific investigation to decipher the neuroprotective and geroprotective mechanism of EISO and its principle components, α- and ß-santalol. Hence the current study was undertaken to assess the protective effects of EISO, and α- and ß-santalol against neurotoxic (6-OHDA/6-hydroxydopamine) and proteotoxic (α-synuclein) stresses in a Caenorhabditis elegans model. Initially, we found that EISO and its principle components exerted an excellent antioxidant and antiapoptotic activity as it was able to extend the lifespan, and inhibit the ROS generation, and germline cell apoptosis in 6-OHDA-intoxicated C. elegans. Further, we showed that supplementation of EISO, and α- and ß-santalol reduced the 6-OHDA and α-synuclein-induced Parkinson's disease associated pathologies and improved the physiological functions. The genetic and reporter gene expression analysis revealed that an EISO, or α- and ß-santalol-mediated protective effect does not appear to rely on DAF-2/DAF-16, but selectively regulates SKN-1 and its downstream targets involved in antioxidant defense and geroprotective processes. Together, our findings indicated that EISO and its principle components are worth exploring further as a candidate redox-based neuroprotectant for the prevention and management of age-related neurological disorders.

Organoruthenium(II) Complexes Ameliorates Oxidative Stress and Impedes the Age Associated Deterioration in Caenorhabditis elegans through JNK-1/DAF-16 Signalling.

New ruthenium(II) complexes were synthesised and characterized by various spectro analytical techniques. The structure of the complexes 3 and 4 has been confirmed by X-ray crystallography. The complexes were subjected to study their anti-oxidant profile and were exhibited significantly greater in vitro DPPH radical scavenging activity than vitamin C. We found that complexes 1-4 confered tolerance to oxidative stress and extend the mean lifespan of mev-1 mutant worms and wild-type Caenorhabditis elegans. Further, mechanistic study and reporter gene expression analysis revealed that Ru(ƞ6-p-cymene) complexes maintained the intracellular redox status and offers stress resistance through activating JNK-1/DAF-16 signaling axis and possibly by other antioxidant response pathway. Notably, complex 3 and 4 ameliorates the polyQ (a Huntington's disease associated protein) mediated proteotoxicity and related behavioural deficits in Huntington's disease models of C. elegans. From these observations, we hope that new Ru(ƞ6-p-cymene) complexes could be further considered as a potential drug to retard aging and age-related neurodegenerative diseases.

Losing its ground: A case study of fast declining populations of a 'least-concern' species, the bonnet macaque (Macaca radiata).

The populations of many species that are widespread and commensal with humans have been drastically declining during the past few decades, but little attention has been paid to their conservation. Here, we report the status of the bonnet macaque, a species that is considered 'least-concern' for conservation. We show that the widely ranging rhesus macaque is expanding its range into the distributional range of the bonnet macaque, a species endemic only to southern India. Bonnet macaques have very low abundance in forests of all types indicating that it is not a typically forest dwelling species. The traditionally preferred habitats of bonnet macaques have been Hindu temples/ tourist spots but our data reveal that nearly 50% population of bonnet macaques has disappeared from such previously occupied spots. Another preferred habitat of bonnet macaques has been roadsides with abundant Ficus trees adjoining croplands. We found that between 2003 and 2015, the roadsides have drastically changed where vegetation has been replaced with barren lands and urbanization. Consequently, the populations of bonnet macaques have declined by more than 65% over the past 25 years, and by more than 50% between 2003 and 2015 alone. We, therefore, conclude that this 'least-concern' species is actually facing serious conservation challenges. We also identify a few places such as small hillocks with natural vegetation and a few temples/tourist spots which are likely to remain stable and thus can serve as 'bonnet macaque conservation reserves'. Since the bonnet macaque shares many traits with several other commensal and 'low-risk' species, it can serve as a model for the development of long-term conservation strategies for most such species.

First Report of the Pale Cyst Nematode, Globodera pallida, in the United States.

In 2006, a cyst nematode was discovered in tare dirt at a potato (Solanum tuberosum) processing facility in eastern Idaho. The nematode was found during a routine survey conducted jointly by the Idaho State Department of Agriculture and the USDA Animal and Plant Health Inspection Service through the Cooperative Agricultural Pest Survey program. Extensive additional sampling from two suspect fields led to the identification of the same nematode in a 45-acre (18.2-ha) field located in northern Bingham County. The morphology of cysts and second-stage juveniles and molecular analyses established the identity of the species as the pale cyst nematode Globodera pallida (Stone 1973) Behrens 1975. Morphological characters used for identification included cyst shape, characteristics of cyst terminal cone including nature of fenestration, cyst wall pattern, anal-vulval distance, number of cuticular ridges between anus and vulva, and Granek's ratio (1,4). The second-stage juvenile morphologies critical for identification were the following: body and stylet length, shape of stylet knobs, shape and length of tail and hyaline tail terminus, and number of refractive bodies in the hyaline part of tail (1,4). Diagnosis as G. pallida was clearly confirmed by two molecular tests. First, PCR-RFLP (restriction fragment length polymorphism) profiles of a ribosomal DNA fragment using restriction enzymes RsaI, TaqI, and AluI (2) were consistent with a G. pallida control and not G. rostochiensis. Second, the ribosomal DNA region that extends from the 3' end of the 18S ribosomal subunit and includes all of ITS1, 5.8S, and ITS2 to the 5' end of the 28S ribosomal subunit was used to generate sequence for the most accurate species determination. Sequences obtained from three individual juveniles were compared with those from several Globodera species (3), revealing unequivocal similarity to G. pallida. This detection represents a new country record for G. pallida in the United States. Collection of additional information regarding distribution of this nematode within the region is underway. References: (1) J. G. Baldwin and M. Mundo-Ocampo. Heteroderinae, Cyst- and Non-cyst-forming Nematodes. Pages 275-362 in: Manual of Agricultural Nematology. W. R. Nickle, ed. Marcel Dekker, New York, 1991. (2) V. C. Blok et al. J. Nematol. 30:262, 1998. (3) L. A. Pylypenko et al. Eur. J. Plant Pathol. 111:39, 2005. (4) A. R. Stone. Nematologica 18:591, 1973.

Health education through non - formal education.

The non-availability of health care in the rural area leads to the problems like infant mortality, infectious disease deaths and malnutrition. Rural health can be promoted both at preventive and promotive levels through non-formal education.