Anticancer property of Bryophyllum pinnata (Lam.) Oken. leaf on human cervical cancer cells

Background: Bryophyllum pinnata (B. pinnata) is a common medicinal plant used in traditional medicine of India and of other countries for curing various infections, bowel diseases, healing wounds and other ailments. However, its anticancer properties are poorly defined. In view of broad spectrum therapeutic potential of B. pinnata we designed a study to examine anti-cancer and anti-Human Papillomavirus (HPV) activities in its leaf extracts and tried to isolate its active principle. Methods: A chloroform extract derived from a bulk of botanically well-characterized pulverized B. pinnata leaves was separated using column chromatography with step-gradient of petroleum ether and ethyl acetate. Fractions were characterized for phyto-chemical compounds by TLC, HPTLC and NMR and Biological activity of the fractions were examined by MTT-based cell viability assay, Electrophoretic Mobility Shift Assay, Northern blotting and assay of apoptosis related proteins by immunoblotting in human cervical cancer cells. Results: Results showed presence of growth inhibitory activity in the crude leaf extracts with IC50 at 552 mu g/ml which resolved to fraction F4 (Petroleum Ether: Ethyl Acetate:: 50: 50) and showed IC50 at 91 mu g/ml. Investigations of anti-viral activity of the extract and its fraction revealed a specific anti-HPV activity on cervical cancer cells as evidenced by downregulation of constitutively active AP1 specific DNA binding activity and suppression of oncogenic c-Fos and c-Jun expression which was accompanied by inhibition of HPV18 transcription. In addition to inhibiting growth, fraction F4 strongly induced apoptosis as evidenced by an increased expression of the pro-apoptotic protein Bax, suppression of the anti-apoptotic molecules Bcl-2, and activation of caspase-3 and cleavage of PARP-1. Phytochemical analysis of fraction F4 by HPTLC and NMR indicated presence of activity that resembled Bryophyllin A. Conclusions: Our study therefore demonstrates presence of anticancer and anti-HPV an activity in B. pinnata leaves that can be further exploited as a potential anticancer, anti-HPV therapeutic for treatment of HPV infection and cervical cancer.

Biochemical characterization of C4 protein of cotton leaf curl Kokhran virus-Dabawali

Background: Cotton leaf curl Kokhran Virus-Dabawali (CLCuKV-Dab) is a monopartite begomovirus encoding two proteins V1 and V2 in the virion sense and four proteins Cl, C2, C3 and C4 in the complementary sense. The C4 protein of monopartite begomoviruses has been implicated to play a role in symptom determination and virus movement. The present work aims at the biochemical characterization of this protein. Methods: The C4 protein of CLCuKV-Dab was purified in fusion with GST and tested for the ability to hydrolyze ATP and other phosphate containing compounds. ATPase activity was assayed by using radiolabeled gamma-32P]-ATP and separating the product of reaction by thin layer chromatography. The hydrolysis of other compounds was monitored by the formation of a blue colored phosphomolybdate complex which was estimated by measuring the absorbance at 655 nm. Results: The purified GST-C4 protein exhibited metal ion dependent ATPase and inorganic pyrophosphatase activities. Deletion of a sequence resembling the catalytic motif present in phosphotyrosine phosphatases resulted in 70% reduction in both the activities. Mutational analysis suggested arginine 13 to be catalytically important for the ATPase and cysteine 8 for the pyrophosphatase activity of GST-C4. Interaction of V2 with GST-C4 resulted in an increase in both the enzymatic activities of GST-C4. Conclusions: The residues important for the enzymatic activities of GST-C4 are present in a motif different from the classical Walker motifs and the non-classical ATP binding motifs reported so far. General significance: The C4 protein of CLCuKV-Dab, a putative natively unfolded protein, exhibits enzymatic activities.

Anti-inflammatory activity of the ethanol extract of Dictamnus dasycarpus leaf in lipopolysaccharide-activated macrophages

Background: Dictamnus dasycarpus is widely used as a traditional remedy for the treatment of eczema, rheumatism, and other inflammatory diseases in Asia. The current study investigates the molecular mechanism of anti-inflammatory action of the ethanol extract of Dictamnus dasycarpus leaf (DE) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: Nitric oxide (NO) production was assessed by Griess reaction and the mRNA and protein expressions of pro inflammatory cytokines, transcription factor, and enzymes were determined by real-time RT-PCR and immunoblotting analysis. Results: DE (0.5 and 1 mg/mL) suppressed the NO production by 10 and 33%, respectively, compared to the untreated group in LPS-stimulated RAW 264.7 cells. DE (0.5 and 1 mg/mL) reduced the mRNA expression of key transcription factor nuclear factor-kappa B by 7 and 24%, respectively compared to the untreated group in LPS activated macrophage. The pro inflammatory cytokines such as tumor necrosis factor a and interleukin 1 beta were also decreased by DE treatment. Moreover, the protein expression of pro inflammatory enzymes, inducible nitric oxide synthase and cyclooxygenase 2 were also dramatically attenuated by DE in a dose dependent manner. Conclusions: These results suggest that Dictamnus dasycarpus leaf has a potent anti-inflammatory activity and can be used for the development of new anti-inflammatory agents.

Spectrophotometric determination of molybdenum with Syzygium jambolanum DC leaf extracts

A new spectrophotometric method for the determination of molybdenum in industrial materials has been developed using the leaf extract of Syzygium jambolanum DC based on the reaction of Mo (VI) at pH 7.0 to produce an orange-yellow complex with an absorption maximum at 426 nm. The molar absorptivity of the complex is 4.27 x 10(4) l mol(-1) cm(-1) and the absorbance, is linear in the range 0.05-0.8 ppm. Sandell sensitivity coefficient was found to be 2.25 x 10(-3) mu g/cm(2). The method is ten times more sensitive than the aqueous thiocyanate system. It has been applied successfully in micronutrient fertilizer, artificial freshwater and sea-water analyses.

Divergence in Patterns of Leaf Growth Polarity Is Associated with the Expression Divergence of miR396

Lateral appendages often show allometric growth with a specific growth polarity along the proximo-distal axis. Studies on leaf growth in model plants have identified a basipetal growth direction with the highest growth rate at the proximal end and progressively lower rates toward the distal end. Although the molecular mechanisms governing such a growth pattern have been studied recently, variation in leaf growth polarity and, therefore, its evolutionary origin remain unknown. By surveying 75 eudicot species, here we report that leaf growth polarity is divergent. Leaf growth in the proximo-distal axis is polar, with more growth arising from either the proximal or the distal end; dispersed with no apparent polarity; or bidirectional, with more growth contributed by the central region and less growth at either end. We further demonstrate that the expression gradient of the miR396-GROWTH-REGULATING FACTOR module strongly correlates with the polarity of leaf growth. Altering the endogenous pattern of miR396 expression in transgenic Arabidopsis thaliana leaves only partially modified the spatial pattern of cell expansion, suggesting that the diverse growth polarities might have evolved via concerted changes in multiple gene regulatory networks.

Assessment Method for Leaf Litters Allelopathic Effect on Cyanobacteria

A new bioassay technique combining leaf disk and softagar over-layer methods was developed to investigate the allelopathic effect of deciduous leaf litters on the growth of cyanobacteria ( Microcystis aeruginosa Kütz.). Bioactive substances exuded from leaf disks caused inhibitory plaques on the agar plate containing cyanobacteria , and the rate of diffusion depended on the specific leaf disk area. Most of the leaf litters collected around reservoirs in Japan showed inhibitory activity to M. aeruginosa , with Rhus trichocarpa Miq., Quercus variabilis Blume and Mallotus japonicus (Thunb.) Muell. Arg. being the strongest among the 22 tested species.(PDF has 4 pages.)

Combining Plant Pathogenic Fungi and the Leaf-Mining Fly, Hydrellia pakistanae, Increases Damage to Hydrilla

Four fungal species, F71PJ Acremonium sp., F531 Cylindrocarpon sp., F542, Botrytis sp., and F964 Fusarium culmorum [Wm. G. Sm.] Sacc. were recovered from hydrilla [ Hydrilla verticillata (L. f.) Royle] shoots or from soil and water surrounding hydrilla growing in ponds and lakes in Florida and shown to be capable of killing hydrilla in a bioassay. The isolates were tested singly and in combination with the leaf-mining fly, Hydrellia pakistanae (Diptera: Ephydridae), for their capability to kill or severely damage hydrilla in a bioassay.

Effects of Triclopyr on Variable-Leaf Watermilfoil

The objective of the study described here was to determine the effect on variable-leaf watermilfoil of various combinations of triclopyr concentrations and exposure times using dosage rates that controlled Eurasian watermilfoil under laboratory and field conditions (Netherland and Getsinger 1992, Getsinger et al. 1997, Petty et al. 1998).

Electrowetting On A Lotus Leaf

Electrowetting on dielectrics has been widely used to manipulate and control microliter or nanoliter liquids in micro-total-analysis systems and laboratory on a chip. We carried out experiments on electrowetting on a lotus leaf, which is quite different from the equipotential plate used in conventional electrowetting. This has not been reported in the past. The lotus leaf is superhydrophobic and a weak conductor, so the droplet can be easily actuated on it through electrical potential gradient. The capillary motion of the droplet was recorded by a high-speed camera. The droplet moved toward the counterelectrode to fulfill the actuation. The actuation speed could be of the order of 10 mm/s. The actuation time is of the order of 10 ms.

Electrowetting on a lotus leaf

Electrowetting on dielectrics has been widely used to manipulate and control microliter or nanoliter liquids in micro-total-analysis systems and laboratory on a chip. We carried out experiments on electrowetting on a lotus leaf which is quite different from the equipotential plate used in conventional electrowetting. This has not been reported in the past. The lotus leaf is superhydrophobic and a weak conductor so the droplet can be easily actuated on it through electrical potential gradient. The capillary motion of the droplet was recorded by a high-speed camera. The droplet moved toward the counterelectrode to fulfill the actuation. The actuation speed could be of the order of 10 mm/s. The actuation time is of the order of 10 ms.

The growth and survival rate of Oreochromis niloticus fries fed (private) with varying percentages of Leucaena leucocephala leaf meal based diets

A-ten-week feeding trial was carried out to evaluate the growth and survival rate of Oreochromis niloticus fed with varying percentage levels of Leucaena leucocephala leaf meal based diets. The substitution rates of L.leucocephala leaf meal for groundnut cake in the various diets were 0% - Diet 1, 25% - Diet 2, 50% - Diet 3 and 75% - Diet 4. Ten fries with an average weight of 0.44g were stocked at the rate of 10 fish per bowl and fed at 5% body weight. Diet 1 with 0% inclusion of Leucaena leaf meal gave a significant difference (P>0.05) in growth and survival rate compared with diets 2, 3 and 4. The water quality parameters recorded were appropriate for fish culture

The growth and survival rate of (private) Oreochromis niloticus fingerlings fed with varying percentages of Leucaena leucocephala leaf meal based diets

A-ten-week feeding trial was carried out to evaluate the growth and survival rate of Oreochromis niloticus fed with varying percentage levels of Leucaena leucocephala leaf meal as a substitute for groundnut cake. The levels in the various diets were 0% - Diet 1, 25% - Diet 2, 50% - Diet 3 and 75% - Diet 4. Ten fingerlings with an average weight of 0.44g were stocked at the rate of 10 fish per bowl and fed at 5% body weight. Diet 1 with 0% inclusion of leucaena leaf meal gave a significant difference (P>0.05) in growth and survival rate compared with diets 2, 3 and 4. The water quality parameters recorded were appropriate for fish culture

Effects of fermented fluted pumpkin leaf (Telfaria occidentalis) on the growth and nutrient utilization of Clarias (Clarias gariepinus)

Clarias (Clarias gariepinus) (Burshell, 1821) fingerlings were fed isonitrogenous diets (38.9% crude protein) with fermented fluted pumpkin leaves (FFPL) replacing different proportion (0,50,75,100%) of extruded soybean meal (ESM) for 8 weeks. Growth responses at the different substitution levels measured. Increasing FFPL intake resulted in better weight gains and higher specific growth rates (SGR) of 0.29, 0.36 and 0.38% per day respectively. The increase in growth from feeding diets containing 75% and 100% of the ESM replaced with FFPL were significantly higher (P<0.05) than those of other diets. Further more fish tissue protein deposition consistently increased with increasing level of FFPL concentration in their diets. Fish fed diets where whole ESM was replace 100% FFPL gave the best overall response in terms of their weight gain, food conversion ratio, protein efficiency ratio, and specific growth rate. Economic considerations indicate the replacement of ESM with FFPL, which is a cheaper ingredient in feeds for Clarias

Estimation of growth and financial analysis through the application of Ipil ipil (Leucaena leucocephala) leaf meal as supplements to soybean and fish meal in the diet of juvenile monosex tilapia (Oreochromis niloticus)

Among plant protein ingredients,ipil ipil (Leucaena leucocephala) leafmeal (ILLM) is considered the most nutritive plant protein source after soybean meal in aquatic feeds. That was proven in a 21-day experiment conducted to assess the response of juvenile Monosex Nile tilapia Oreochromis niloticus with four iso-nitrogenous formulated diets: One control diet was formulated based on fishmeal, one on soybean meal and one on rice bran, ipil ipil leafmeal was also included in experimental diets.