Pharmaceutical characterization and exploration of Arkeshwara rasa in MDA-MB-231 cells.

BACKGROUND: The diverse specificity mode of cancer treatment targets and chemo resistance demands the necessity of drug entities which can address the devastating dynamicity of the disease. OBJECTIVES: To check the anti-tumour potential of traditional medicine rich in polyherbal components and metal nanoparticle namely Arkeshwara rasa (AR). MATERIAL METHODS: The AR was prepared in a modified version with reference from Rasaratna Samuchaya and characterized using sophisticated instrumental analysis including XRD, SEM-EDAX, TEM, TGA-DSC, and LC-MS and tested against the MDA-MB-231 cell line to screen cell viability and the cytotoxicity with MTT, SRB and the AO assay. RESULTS: XRD pattern shows cubic tetrahedrite structure with Sb, Cu, S peaks and trace elements like Fe, Mg, etc. The particle size of AR ranges between 20 and 30 nm. The TGA points thermal decomposition at 210 °C and the metal sulphide peaks in DSC. LC-MS analysis reveals the components of the formulation more on the flavonoid portion. The IC50 value of MTT and SRB are 25.28 µg/mL and 31.7 µg/mL respectively. The AO colorimeter substantiated the cell viability and the apoptosis figures of the same cell line. The AR exhibits cytotoxicity and reaffirms the apoptosis fraction with SRB assay. CONCLUSIONS: The Hesperidine, Neohesperidin, Rutin components in the phytochemical pool can synergize the anti-tumour potential with either influencing cellular pathways or decreasing chemo resistance to conventional treatment. AR need to be further experimented with reverse transcription, flow cytometry, western blotting, etc.

Low grade cirrhosis induces cognitive impairment and motor dysfunction in rats: could be a model for minimal hepatic encephalopathy.

Hepatic encephalopathy (HE) is a nervous system disorder developed in the patients with liver cirrhosis. The low grade cirrhosis is of common occurrence, however, whether and the extent to which it affects brain function is not clearly understood. The present article examines certain neurobehavioral parameters in the rats with low grade chronic liver failure (CLF) induced by intraperitoneal administration of 50mg/kgb.w. thioacetamide up to 14 days. During Morris Water Maze tasks, the CLF rats, as compared to the control, showed insignificant decline in the escape latency score to find out the hidden platform throughout the learning days and also stayed for a significantly declined (p<0.01) time period at the place of the hidden platform during the retrieval test. They also showed impairment in the conditional discrimination ability, reflected by a significant decline in the active avoidance score (p<0.05) and increment in the number of non-response (p<0.05) during shuttle box tests. On rotarod performance, they exhibited significant decline in their riding time (p<0.01-001) on the rotating rod as a function of increasing speed. The findings suggest a moderate level cognitive impairment and motor dysfunction in the low grade CLF rats. Since, these impairments correlate with the early stage manifestation of HE in the patients, these CLF rats could serve as a model to study the pathogenesis of minimal hepatic encephalopathy.

One pot synthesis of Cu(II) 2,2'-bipyridyl complexes of 5-hydroxy-hydurilic acid and alloxanic acid: synthesis, crystal structure, chemical nuclease activity and cytotoxicity.

A barbiturate derivative [1,5-dihydro-5-[5-pyrimidine-2,4(1H,3H)-dionyl]-2H-chromeno[2,3-d] pyrimidine-2,4(3H)-dione] (LH(4)) was allowed to react with 2,2'-bipyridyl-dinitrato-Copper(II)-dihydrate which provides two complexes, characterized as [Cu(bpy)(L1)]·3H(2)O (1) and [Cu(bpy)(L2)]·H(2)O (2), where bpy = 2,2'-bipyridine, L1 = 5-hydroxy-hydurilic acid and L2 = alloxanic acid. In a separate reaction of LH(4) with Cu(NO(3))(2)·H(2)O another type of complex [Cu(LH(3))(2)·(H(2)O)(2)]·4H(2)O (3) is formed. The complexes were characterized by single crystal X-ray crystallography, physicochemical and electrochemical studies. The interaction of complexes 1 and 3 with DNA was monitored using absorption and emission titrations as well as circular dichroism spectroscopy. The complexes were found to cleave supercoiled plasmid DNA to nicked circular and linear DNA. Complexes 1 and 3 were also tested against T-cell lymphoma (Dalton lymphoma DL) and showed significant cytotoxic activity with IC(50) values of ~9.0 nM and 0.6 nM.

Anti-HIV and cytotoxic ruthenium(II) complexes containing flavones: biochemical evaluation in mice.

Ru(II) polypyridyl complexes containing 3-hydroxyflavone derivatives as coligands were screened for anti-HIV and cytotoxic activities against eleven tumor cell lines. In order to check the effect of flavones containing Ru(II) complexes in vivo on a mammal, a representative complex Ru(L)2(DMSO)2 x 5H2O (LH-3-Hdroxy-4'-benzyloxyflavone; M5) was orally administered to adult male mice. Its effects on protein content and LDH were studied in different tissues of the animal. The compound got absorbed and retained in the blood between 1-3 hr after feeding. As compared to the normal and DMSO control sets, tissue specific significant reversible changes in the protein content as well as in LDH activity were observed between 1-4 hr of treatment. However, on polyacrylamide gel electrophoresis, except some tissue specific transitory alterations, expression patterns of five LDH isozymes were unchanged after feeding the compound. The present results suggested that in addition to its potent cytotoxic and anti-HIV effects on cell lines in vitro, M5 inhibited LDH activity, but reversibly with a little effect on biosynthetic status of the enzyme in mice.

Identification of cells expressing a D type G1 cyclin in matured brain: implication for its role in neuronal function.

The multiple cyclins and their catalytic subunit, cdc2-related kinase, play essential roles in the eucaryotic cell cycle. To examine their potential roles in highly differentiated nervous system, we determined their activity and localization in rat brain. p13suc1 associated histone H1 kinase activity was dramatically increased at the onset of brain maturation. The increased kinase activity was coprecipitated with cyclin D1, a type of G1 cyclin, which also increased with brain maturation. Immunohistochemical analysis demonstrated that anti-cyclin D1-like immunoreactivity was exclusively localized to neurons. From these findings, it is suggested that a type of cdc2-related kinase regulated by cyclin D1 might function in neurons independent of cell cycle progression.

Age-linked alterations in fructose-2,6-bisphosphate-induced modulation of rat muscle phosphofructokinase.

Native muscle phosphofructokinase (PFK: EC 2.7.1.11) isolated from 25- and 100-week-old rats was subjected to in vitro studies on fructose-2,6-bisphosphate-induced alterations in the regulatory roles of other key metabolic modulators of this enzyme. Although fructose 2,6-bisphosphate-mediated reversal of citrate inhibition did not show any age-related difference, synergism with glucose-1,6-bisphosphate effect was found to be slightly increased with the enzyme of 100-week-old rats. In addition, apart from a significant decrease in the extent of fructose-2,6-bisphosphate activation, synergism with AMP activation and reversal of ATP and pyridoxal-5-phosphate inhibitions were observed to be decreased markedly with the enzyme of 100-week-old rats in comparison with that of 25-week-old rats. Such age-dependent alterations in muscle PFK provide evidence for conformational modification in this enzyme as a function of age.

Evidence for tissue specific alterations in Zn2+-induced conformational changes in fructose-bisphosphatase of senescent rats.

In vitro studies on Zn2+-induced modulations in certain allosteric control of fructose-1,6-bisphosphatase (FBPase: EC 3.1.3.11), isolated from liver and muscle of 28- and 97-wk old rats were carried out in parallel. Similar chromatographic elution on ion-exchanger and electrophoretic mobility on polyacrylamide gels revealed similarity in charge and molecular size of the enzyme proteins from the two ages of rats. Regarding Zn2+ induced modulations, almost all the parameters used did not show any age-dependent significant alteration with liver enzyme. However, in case of muscle FBPase, apart from a significant increase in Ki for ZnCl2, Zn2+-induced modulations in substrate affinity and AMP inhibition were observed to be altered markedly with the enzyme of 97-wk-old rats in comparison to that of 28-wk-old rats. Thus, it suggests age-associated alterations in Zn2+-mediated conformational modification in the muscle enzyme. This has been further supported by tissue-specific usual pattern of substrate affinity in the absence of Zn2+ and exhibition of normal AMP inhibition after replacement of Zn2+ by EDTA. Such age-dependent changes induced by Zn2+ in muscle FBPase may be of high physiological significance with advancing age of the animal.

Changes in the modulations of kinetics and allosteric properties of muscle phosphofructokinase of young and old rats.

In vitro studies on various modulations in kinetics and allosteric properties of muscle phosphofructokinase (EC 2.7.1.11) were undertaken, using purified enzyme from young (25-weeks) and old (100-weeks) albino rats. In comparison to normal K0.5 values for fructose-6-phosphate, increase in this value in response to ATP and citrate inhibition, decrease in K0.5 due to AMP activation and extent of ATP inhibition with increase in pH, were observed to be decreased more markedly with the enzyme of old than with that of young rats. Extent of citrate inhibition, reversals of ATP and citrate effects in response to AMP activation, and synergism of citrate and ATP inhibitions were also seen to be decreased considerably with muscle PFK of old in comparison to that of young rats. Such age-related changes in muscle PFK suggest the alterations in allosteric regulation of this enzyme during aging of the animal.

Alterations in the sensitivity to endogenous proteolysis due to storage of fructose-1,6-bisphosphatase extracts of liver and muscle of young and old rats.

The sensitivity of fructose-1,6-bisphosphatase (FBPase: EC 3.1.3.11) to endogenous proteolysis was studied in liver and muscle extracts of young (28 weeks) and old (97 weeks) rats (stored at 4 degrees +/- 2 degrees C) by monitoring the alterations in its properties. Apart from the level of total enzyme activity, decrease in activity ratio (at pH 7.0/9.2) and sensitivity to AMP inhibition no age-dependent significant change was found during storage of the liver extract. However, a more significant increase in activity, comparatively less decrease in activity ratio, and retention of sensitivity to AMP inhibition were observed during the storage of muscle enzyme extract from old rats than from young rats. Using gel electrophoresis, an extra degradative active product was observed in all samples except those of old rat muscle. These results may indicate tissue-specific alterations in the susceptibility of FBPase to endogenous proteolysis during old age in the rat.