In vitro antioxidant and cytotoxicity studies of Curcuma

Abstract: Curcuma amada Roxb, of Zingiberaceae is an important active spice used in traditional system of medicine. They also used in the manufacture of pickles, chutney, salad and jam. Phosphomolybdenam method was used to measure the total antioxidant capacity (TAC) of C. amada ethanolic rhizome extract (CAEREt). The free radical scavenging capacity of CAEREt was also carried out by ABTS and DPPH methods. The cytotoxicity effect of CAEREt was determined by Trypan blue dye exclusion method and MTT assay using two different cell lines such as Dalton Lyphoma Ascites (DLA) and Human Cervical Cancer Cell Line (HeLa). The results showed that the rhizome extract of C. amada rich in phytocompounds and have exhibited better antioxidant and cytotoxicity properties

Hypoglycaemic and Hypolipidaemic Effects of Withania somnifera Root and Leaf Extracts on Alloxan-Induced Diabetic Rats

Withania somnifera is an important medicinal plant, which is used in traditional medicine to cure many diseases. Flavonoids were determined in the extracts of W. somnifera root (WSREt) and leaf (WSLEt). The amounts of total flavonoids found in WSREt and WSLEt were 530 and 520 mg/100 g dry weight (DW), respectively. Hypoglycaemic and hypolipidaemic effects of WSREt and WSLEt were also investigated in alloxan-induced diabetic rats. WSREt and WSLEt and the standard drug glibenclamide were orally administered daily to diabetic rats for eight weeks. After the treatment period, urine sugar, blood glucose, haemoglobin (Hb), glycosylated haemoglobin (HbA1C), liver glycogen, serum and tissues lipids, serum and tissues proteins, liver glucose-6-phosphatase (G6P) and serum enzymes like aspartate transaminase (AST), alanine transaminase (ALT), acid phosphatase (ACP) and alkaline phosphatase (ALP) levels were determined. The levels of urine sugar, blood glucose, HbA1C, G6P, AST, ALT, ACP, ALP, serum lipids except high density lipoprotein-bound cholesterol (HDL-c) and tissues like liver, kidney and heart lipids were significantly (p < 0.05) increased, however Hb, total protein, albumin, albumin:globulin (A:G) ratio, tissues protein and glycogen were significantly (p < 0.05) decreased in alloxan-induced diabetic rats. Treatment of the diabetic rats with WSREt, WSLEt and glibenclamide restored the changes of the above parameters to their normal level after eight weeks of treatment, indicating that WSREt and WSLEt possess hypoglycaemic and hypolipidaemic activities in alloxan-induced diabetes mellitus (DM) rats

Determination of Mineral Content in Methanolic Safflower (Carthamus tinctorius L.) Seed Extract and Its Effect on Osteoblast Markers

Safflower (Carthamus tinctorius L.) seeds are used as a folk medicine to enhance bone formation or to prevent osteoporosis in Korea. Therefore, the methanolic extract of safflower seeds (MESS) containing high mineral content, such as calcium (Ca), potassium (K) and phosphorous (P), was evaluated for the role on osteoblast (Ob) markers of Sprague-Dawley rats. In serum of 3 to 11 weeks (wks) old rats, both osteocalcin (OC) content and bone-specific alkaline phosphatase (B-ALP) activity increased to their maximum levels in 4–7 wks. Hence, 3 wks old rats were selected for 8 wks oral treatment of MESS, resulted in the significant increase of Ob markers in serum such as OC content (4–8 wks), B-ALP activity (1–2 wks) and insulin-like growth factor I (IGF-I) level (1 wk), and the growth parameter such as the length of femur (2–8 wks) and tibia (4 wks). On the basis of Pearson’s correlation coefficient, there were a moderate correlation between OC and B-ALP at 8 wks, a low correlation between OC and IGF-I at 1, 4 and 8 wks, a moderate correlation between OC and femur length at 1, 2 and 8 wks, and a moderate correlations between OC and tibia length at 1 and 8 wks of MESS-treated groups. The result reveals that the changes of OC correlated at low to moderate level with the changes of B-ALP activity, IGF-I content and femur and tibia length in the MESS-treatment period. On the other hand, there were a strong correlation between IGF-I and femur length at 2 wks and moderate correlation between IGF-I and tibia length at 1, 2 and 8 wks of MESS-treated groups. Therefore, the effect of MESS on bone formation likely appears to be mediated by IGF-I at the early stage of treatment

Phytochemical content of leaf and stem of Marsilea quadrifolia (L.)

The present study was aimed to screen and quantify the phytochemicals by qualitative and quantitative analysis in methanol and aqueous leaf and stem extracts of Marsilea quadrifolia(L.). In qualitative analysis, the phytochemical compounds such as tannins, saponins, flavonoids, steroids, terpenoids, triterpenoids, alkaloids, carbohydrates, proteins, anthroquinones, phenolic compounds and phytosterol were screened. Among these phytocompounds tannins, saponins, flavonoids, steroids, alkaloids, carbohydrates, proteins and phenolic compounds were observed in methanol and aqueous leaf and stem extracts of M. quadrifolia. Anthroquinones were absent in both leaf and stem extracts of M. quadrifolia. The content of phenolic compounds 8.34&plusmn;0.92 mg/g and 7.31&plusmn;0.46 mg/g, flavonoids 7.46&plusmn;0.64 mg/g and 6.45&plusmn;0.68 mg/g, alkaloids 6.12&plusmn;0.51 mg/g and 5.89&plusmn;0.61 mg/g, tannins 6.58&plusmn;0.72 mg/g and 6.07&plusmn;0.56 mg/g and saponins 5.32&plusmn;0.48 mg/g and 6.30&plusmn;0.58 mg/g were determined in leaf and stem of M. quadrifolia, respectively. So, the present study confirmed that the presence of phytocompounds in leaf and stem of M. quadrifolia

Phytochemical Analysis and Nematicidal Activity of Ethanolic Leaf Extracts of Datura metel, Datura innoxia and Brugmansia suaveolens Against Meloidogyne incognita

The present study was aimed to screen the phytochemicals and quantification of alkaloids, phenolic compounds and flavonoids and to evaluate the nematicidal activity of ethanolic leaf extracts of Datura metel, Datura innoxia and Brugmansia suaveolens against Meloidogyne incognita. Phytochemical screening of leaf extracts of D. metel, D. innoxia and B. suaveolens was carried out by qualitative analysis and the results revealed that the presence of bioactive compounds like alkaloids, steroids, flavonoids, terpenoids, phenolic compounds, tannins, anthroquinone glycosides, saponins and triterpenes. The quantitative phytochemical analysis showed that the maximum amount of alkaloids 12.12 mg/g in leaf of D. innoxia, phenolic compounds 5.675 mg/g in leaf of D. metel and flavonoids 4.945 mg/g in leaf of B. suaveolens. The nematicidal activity of extracts of D. metel, D. innoxia and B. suaveolens were studied against the root knot nematode M. incognita. For nematicidal activity, the leaf extracts were used at different concentrations like 250 ppm, 500 ppm, 750 ppm and 1000 ppm in different exposure time like 24 hrs, 48 hrs and 72 hrs. The in vitro nematicidal activity results suggested that the extracts of D. metel, D. innoxia and B. suaveolens were possessed nematicidal activity. From this study, we concluded that the extracts of D. metel, D. innoxia and B. suaveolens contain phytochemicals and the plants were exhibited nematicidal activity against root knot nematode M. incognita. The present study may be useful for the identification and isolation of novel active nematicidal compounds from the leaf of D. metel, D. innoxia and B. suaveolens

Expression of Aspergillus nidulans phy Gene in Nicotiana benthamiana Produces Active Phytase with Broad Specificities

A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs