A withanolide coagulin-L inhibits adipogenesis modulating Wnt/ß-catenin pathway and cell cycle in mitotic clonal expansion.

Obesity is a result of adipocyte hypertrophy followed by hyperplasia. It is a risk factor for several metabolic disorders such as dyslipidemia, type-2 diabetes, hypertension, and cardiovascular diseases. Coagulanolides, particularly coagulin-L isolated from W. coagulan has earlier been reported for anti-hyperglycemic activity. In this study, we investigated the effect of coagulin-L on in vitro models of adipocyte differentiation including 3T3-L1 pre-adipocyte, mouse stromal mesenchymal C3H10T1/2 cells and bone marrow derived human mesenchymal stem cells (hMSCs). Our results showed that, coagulin-L reduces the expressions of peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer-binding protein α (C/EBPα), the major transcription factors orchestrating adipocyte differentiation. Detailed analysis further proved that early exposure of coagulin-L is sufficient to cause significant inhibition during adipogenesis. Coagulin-L inhibited mitotic clonal expansion (MCE) by delayed entry in G1 to S phase transition and S-phase arrest. This MCE blockade was caused apparently by decreased phosphorylation of C/EBPß, modulation in expression of cell cycle regulatory proteins, and upregulation of Wnt/ß-catenin pathway, the early stage regulatory proteins of adipogenic induction. Taken together all evidences, a known anti-hyperglycemic agent coagulin-L has shown potential to inhibit adipogenesis significantly, which can be therapeutically exploited for treatment of obesity and metabolic syndrome.

Antifilarial activity of marine sponge Haliclona oculata against experimental Brugia malayi infection.

The present study incorporates the findings on in vitro and in vivo antifilarial activity in the marine sponge, Haliclona oculata using an experimental rodent infection of human lymphatic filarial parasite, Brugia malayi. The in vitro antifilarial action was determined on both adult female worms as well as microfilariae using two parameters viz. adverse effect on motility and inhibition in MTT reduction by the treated adult parasite over control worm. The antifilarial activity could be located in the methanol extract and one of its four fractions (chloroform). Bioactivity guided fractionation of chloroform fraction led to localization of in vitro activity in one of its eight chromatographic fractions. Methanol extract, chloroform fraction and one of the chromatographic fractions revealed IC(50) values of 5.00, 1.80, and 1.62µg/ml, respectively when adult B. malayi were exposed to these test samples for 72h at 37°C. Under similar exposure conditions, the IC(50) values for microfilariae were 1.88, 1.72 and 1.19µg/ml, respectively. The active test samples were found to be safe revealing >10 selectivity indices (SI) on the basis of cytotoxicity to Vero cells (monkey kidney cells) and therefore selected for in vivo evaluation against primary (adult B. malayi intraperitoneal transplanted jird) and secondary (subcutaneous infective larvae induced mastomys) screens. In primary jird model, the three test samples at 100mg/kg for five consecutive days by subcutaneous route demonstrated significant macrofilaricidal efficacy to the tune of 51.3%, 64% and 70.7% by methanol extract, chloroform and chromatographic fraction, respectively. The three samples demonstrated 45-50% macrofilaricidal activity with moderate embryostatic effect in secondary model at 5×500, 5×250 and 5×125mg/kg by oral route. Chromatographic fraction possessing highest antifilarial action was primarily found to be a mixture of four alkaloids Mimosamycin, Xestospongin-C, Xestospongin-D and Araguspongin-C in addition to few minor compounds.

Cassane diterpenes from Caesalpinia bonduc.

Three cassane diterpene hemiketals, caesalpinolide-C, caesalpinolide-D, caesalpinolide-E and one cassane furanoditerpene were isolated from Caesalpinia bonduc. The molecular structures were elucidated using NMR spectroscopy in combination with IR, UV and mass spectral data and relative stereochemistries were determined through ROESY correlation. The isolated compounds were tested for their antiproliferative activity against MCF-7 (breast adenocarcinoma), DU145 (prostate carcinoma), C33A (Cervical carcinoma) and Vero (African green monkey kidney fibroblast) cells.

Bioactivity of marine organisms: part X--screening of some marine fauna from the Indian coasts.

Methanolic and chloroform-methanol extracts of 25 identified species of marine fauna have been screened for a wide range of biological activities. Of these, 2 extracts exhibited diuretic activity, while antibacterial, antiviral, oxytocic and spasmolytic activities, were observed in 1 extract each.

Bioactivity of marine organisms: Part IX--screening of some marine flora from the Indian coasts.

Alcoholic extracts of 48 identified species of marine flora were screened for a wide range of biological activities. Of these, 3 extracts showed diuretic activity while 2 extracts showed hypotensive effect.

Systematic and ecological studies on Chlorophyceae of North India and their relationship with water quality.

In the course of systematic and ecological studies on algal flora of fresh water environment of three different agroclimatic zone of Uttar Pradesh revealed one hundred eighty two species represented by fifty-two genera inhabiting fresh water bodies having different physico-chemical properties. In both the regions members of order Conjugales were dominant and represented by ninety nine species belonging to fourteen genera. This is followed by Chlorococcales having fifty two species represented by nineteen genera and Chaetophorales with nine species of four genera only. In the central Uttar Pradesh a positive correlation was found between hydrogen ions concentration with temperature and species diversity, while in western Uttar Pradesh a highly positive correlation was found in electrical conductivity and total dissolved solids.

Tagging and mapping of a rice gall midge resistance gene, Gm8, and development of SCARs for use in marker-aided selection and gene pyramiding.

Using amplified fragment length polymorphisms (AFLPs) and random amplified polymorphic DNAs (RAPDs), we have tagged and mapped Gm8, a gene conferring resistance to the rice gall midge (Orseolia oryzae), a major insect pest of rice, onto rice chromosome 8. Using AFLPs, two fragments, AR257 and AS168, were identified that were linked to the resistant and susceptible phenotypes, respectively. Another resistant phenotype-specific marker, AP19(587), was also identified using RAPDs. SCAR primers based on the sequence of the fragments AR257 and AS168 failed to reveal polymorphism between the resistant and the susceptible parents. However, PCR using primers based on the regions flanking AR257 revealed polymorphism that was phenotype-specific. In contrast, PCR carried out using primers flanking the susceptible phenotype-associated fragment AS168 produced a monomorphic fragment. Restriction digestion of these monomorphic fragments revealed polymorphism between the susceptible and resistant parents. Nucleotide BLAST searches revealed that the three fragments show strong homology to rice PAC and BAC clones that formed a contig representing the short arm of chromosome 8. PCR amplification using the above-mentioned primers on a larger population, derived from a cross between two indica rice varieties, Jhitpiti (resistant parent) and TN1 (susceptible parent), showed that there is a tight linkage between the markers and the Gm8 locus. These markers, therefore, have potential for use in marker-aided selection and pyramiding of Gm8 along with other previously tagged gall midge resistance genes [ Gm2, Gm4(t), and Gm7].

The antifilarial activity of a marine red alga, Botryocladia leptopoda, against experimental infections with animal and human filariae.

The antifilarial activity of the marine red alga Botryocladia leptopoda against rodent and human lymphatic filarial parasites is described. The animal filarial species included Litomosoides sigmodontis and Acanthocheilonema viteae maintained in cotton rats and Mastomys coucha, respectively, while a subperiodic strain of the human lymphatic filarial parasite Brugia malayi was maintained in M. coucha. The crude extract and its hexane fraction brought about a marked reduction in the peripheral microfilarial level in both of the rodent filarial parasites L. sigmodontis and A. viteae. The microfilaricidal effect began slowly from day 8 or 15 after initiation of treatment and increased with time with a very high efficacy at the end of the observation period against both rodent filariids. The microfilaricidal efficacy was, however, not as prominent in the case of B. malayi. The antifilarial activity, which occurred in the hexane fraction, exerted action at a much lower dose. The product killed a significant proportion of A. viteae and L. sigmodontis adult parasites. In the case of B. malayi, although the macrofilaricidal efficacy was much less than that of the rodent parasites, it (hexane fraction) caused sterilization of a significant proportion of the surviving female parasites. The present findings indicate the possibility of developing an adulticidal and female sterilizing agent against filarial parasites from a marine red alga.

Bioactivity of marine organisms: Part VIII--Screening of some marine flora of western coast of India.

Methanolic extracts of 31 botanically identified species of marine flora, collected from Gujarat Coast, have been screened for a wide range of biological activities. Of these, 3 extracts showed anti-implantation, 2 had antiviral, 2 showed hypotensive, 1 had anti-inflammatory while 12 extracts showed diuretic activities. The antiviral activity; against EMCV, was confirmed in one alga. The active principles and results of these studies are reported.

Chalcone synthase in rice (Oryza sativa L.): detection of the CHS protein in seedlings and molecular mapping of the chs locus.

The chalcone synthase is a key enzyme that catalyses the first dedicated reaction of the flavonoid pathway in higher plants. The chs gene and its protein product in rice has been investigated. The presence of a chalcone synthase (CHS) protein in rice seedlings and its developmental stage-specific expression has been demonstrated by western analysis. The chalcone synthase of rice was found to be immunologically similar to that of maize. A rice cDNA clone, Os-chs cDNA, encoding chalcone synthase, isolated from a leaf cDNA library of an indica rice variety Purpleputtu has been mapped to the centromeric region of chromosome 11 of rice. It was mapped between RFLP markers RG2 and RG103. RG2 is the nearest RFLP marker located at a genetic distance of 3.3 cM. Some segments of chromosome 11 of rice including chs locus are conserved on chromosome 4 of maize. The markers, including chs locus on chromosome 11 of rice are located, though not in the same order, on chromosome 4 of maize. Genetic analysis of purple pigmentation in two rice lines, Abhaya and Shyamala, used in the present mapping studies, indicated the involvement of three genes, one of which has been identified as a dominant inhibitor of leaf pigmentation. The Os-chs cDNA shows extensive sequence homology, both for DNA and protein (deduced), to that of maize, barley and also to different monocots and dicots.

PCR-based DNA markers linked to a gall midge resistance gene, Gm4t, has potential for marker-aided selection in rice.

Rice DNAs from a gall midge resistant variety, 'Abhaya', a susceptible variety, 'Tulsi' and their F3 progeny were screened using 500 random primers in conjunction with bulked-segregant analysis in a polymerase chain reaction (PCR) with a view to detecting random amplified polymorphic DNAs (RAPDs) linked to the gene, Gm4t, which confers resistance to gall midge, a dipteran insect pest of rice. A total of 454 primers were able to produce a distinct amplification pattern, and 3695 bands/loci were amplified between the phenotypically different parents. Of these, 304 bands were polymorphic between the parents, with 19 being phenotypespecific. One of these primers, E20, amplified 2 bands, E20570 and E20583, which are tightly linked to resistance and susceptibility, respectively. These specific bands were cloned and sequenced, and a 94% sequence homology was found between the two fragments. Two specific 20-mer oligonucleotides were synthesized, based on the sequence information of E20583, for use in PCR amplification directly from genomic DNAs. These PCR primers were able to amplify phenotype-specific bands, a 583-bp fragment in susceptible F3 lines and a 570-bp fragment in resistant F3 lines that had been derived from a cross between the parents, indicating their potential and utility for marker-aided selection of the Gm4t gene in rice. Its use would facilitate the early and efficient selection of resistant genes in plant breeding programmes and even in those areas where the insect is not known to occur. These phenotype-specific bands are single-copy sequences and are being mapped to ascertain their chromosomal location in rice.

Bioactivity of marine organisms: Part VI--Screening of some marine flora from Indian coasts.

Alcoholic extracts of 50 botanically identified species of marine flora have been screened for a wide range of biological activities. Of these, 2 extracts exhibited anti-amoebic and antiviral activity each, 3 of them had anti-implantation activity; 9 had hypoglycaemic activity while hypotensive activity was associated with 11 extracts; 14 extracts were found to be diuretic and 1 of them had anti-inflammatory activity. Further, 10 of these extracts exhibited 2 types of activities while a combination of 3 and 4 types of activities was observed in one extract each. Follow-up studies have been carried out in some plants with confirmed activity. The active principles and results of these studies are reported.

A note on etno - oceano biology of andaman and nicobar islands.

The present paper coins a new term Ethno-Oceanobiology, a new sub-discipline of ethnobiology and deals with the uses of some marine plants by tribals of Andaman and Nicobar Islands.

Stability constants and thermodynamic functions of molybdenum and uranium chelates formed with dl-alpha-aminobutyric acid.

The stability constants and thermodynamic functions involved in the formation of Mo(VI) and U(VI) chelates with dl-alpha-aminobutyric acid have been determined potentiometrically. It is observed that in the case of Mo(VI) system three chelates ML, ML(2) and ML(3) are formed stepwise, whereas in the U(VI) system only two chelates ML and ML(2) are formed before precipitation occurs, and both steps occur almost simultaneously. Results show that entropy makes a predominant contribution to the stability of both Mo(VI) and U(VI) chelates.

HYSTERIA OR HYPOGLYCAEMIA (A case Report of Insulinoma).

A case of Insulinoma is presented here because of the uncommon and peculiar clinical manifestation in the form of fits of unconsciousness with bizzare movements of limbs, which were considered to be due to Conversion Reaction and treated as such for a period of two years without any success. She recovered and was relieved of symptoms after surgery.

Potentiometric determination of stepwise stability constants of zirconium, thorium and uranium chelates of asparagine and glutamine.

The metal chelates of Zr(IV), Th(IV) and U(VI) with asparagine and glutamine have been studied potentiometrically. Stepwise stability constants in 0.1 M sodium perchlorate at 25 degrees are as follows. Asparaginate chelates-log K(1) 8.80, log K(2) 6.25 for Zr, log K(1) 6.79, log K(2) 6.16 for U, log K(1) 8.28, log K(2) 7.77 and log K(3) 7.72 for Th. Glutaminate chelates-log K(1) 8.75, log K(2) 6.10 for Zr, log K(1) 6.63, log K(2) 6.22 for U, log K(1) 8.30, log K(2) 7.61 and log K(3) 7.55 for Th.

Potentiometric determination of stepwise stability constants of vanadium, molybdenum and tungsten chelates formed with asparagine and glutamine.

The metal chelates of V(IV), Mo(VI) and W(VI) formed with asparagine and glutamine have been studied potentiometrically. Stepwise stability constants in O.1M sodium perchlorate at 25 degrees C are reported as follows. Asparagine chelates--log K(1)K(2) 14.50 and log K(3) 4.04 for V, log K(1) 8.06, log K(2) 7.29 and log K(3) 3.45 for Mo, and log K(1) 5.84, log K(2) 5.11 and log K(3) 3.30 for W. Glutamine chelates--log K(1)K(2) 14.45, and log K(3) 4.07 for V, log K(1) 7.90, log K(2) 6.93, and log K(3) 3.35 for Mo, and log K(1) 5.76, log K(2) 5.09 and log K(3) 3.20 for W.

Potentiometric determination of stepwise stability constants of zirconium and thorium chelates formed with aspartic and glutamic acids.

The metal chelates of Zr(IV) and Th(IV) with aspartic and glutamic acids have been studied potentiometrically. Stepwise stability constants in 0.1 M sodium perchlorate at 25 degrees are reported as: aspartate chelates-log K(1) 9.70, log K(2) 6.85, and log K(3) 3.50 for Zr, and log k(1) 9.23, log K(2) 8.57, log K(3) 4.55 and log K(4) 3.87 for Th; glutamate chelates-log K(1) 9.60, log K(2) 6.40 and log K(3) 3.32 for Zr and log K(1) 9.11, log K(2) 8.52, log K(3) 4.18 and log K(4) 3.62 for Th.