CLEISTANTHIN B SHOWS A POTENT CYTOTOXIC ACTIVITY AGAINST COLORECTAL CANCER CELLS

Objective: The aim was to find out the cleistanthin B sensitive cancer cell type among a panel of cancer cell lines. Methods: The 50% inhibitory concentrations (IC50) of cleistanthin B against different cancer cells were determined by 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide assay. The cell death caused by cleistanthin B in colorectal cancer (CRC) cells was evaluated by acridine orange and ethidium bromide (AO-EB) dual staining. Using short exposure, we generated the 5-fluorouracil</AQ1> and oxaliplatin (5-FU+Ox) surviving cells from the parental HT-29 CRC cell lines. These surviving CRC cells were further treated with cleistanthin B either alone or combined with 5-FU. Annexin V apoptosis assay was used to determine the combined effect of cleistanthin B with 5-FU against HT-29 cells. Results: The IC50 values of cleistanthin B were found to be 3.6±0.55, 5.2±0.51, 8.6±1.02, 10.5±1.50, 18.3±3.71, 25.8±5.50, and 26.7±5.90 μg/mL against HT-29, SW-480, HCT-15, HELA, MDA-MB-231, A549, and DU145, respectively. The IC50 value of cleistanthin B against L132 cells was >100 μg/mL. The cleistanthin B treated HT-29, SW-480, and HCT-15 CRC cells showed apoptotic changes such as chromatin condensation, nuclear fragmentation, and formation of apoptotic bodies in the AO-EB dual staining method. Flow cytometry analysis revealed that cleistanthin B enhances the 5-FU induced apoptosis against 5-FU+Ox surviving HT-29 CRC cells. Conclusion: Cleistanthin B is relatively more potent against CRC cells than other cancer cells, and it induces apoptosis mediated cell death in CRC cells. Cleistanthin B enhances the anticancer activity of 5-FU against HT-29 CRC cells

Association between placental vitamin D receptor expression and cord blood vitamin D level and its effect on the birth weight of newborns

Background: Vitamin D deficiency during pregnancy affects fetal growth and development. The present study explored the association between vitamin D levels in cord blood and placental vitamin D receptor expression.Methods: A total of 54 subjects with live-born singleton deliveries were included in SRM Medical College Hospital and Research Centre, Kattankulathur, India. Cord blood and placenta was obtained at the time of delivery.      Results: The mean newborn vitamin D level was 20.07 ng/mL (13-27.50). The mean of relative vitamin D receptor (VDR) expression was 1.45Ct (range 0.10-2.6). Mean comparison of cord serum 25(OH) D3 and VDR revealed significant at p<0.05 by student’s t-test. The newborn anthropometric measurement of head circumference 31.44±3.99 cm, Chest circumference 28.88±4.30 cm, Body length 45.52±8.05 cm, Birth weight 2660.61±353.27 g had significant association with mean VDR expression at p <0.05 level by student t-test analysis. Further, Pearson correlation coefficient also revealed strong positive correlation of placental VDR and cord serum vitamin D (r=0.853, p <0.05).Conclusions: The present study indicates that there was a high prevalence of vitamin D deficiency in the south Indian newborns and the study suggested that VDR had crucial role in maternal to fetal nutrient transfer mechanism. Vitamin D status in cord blood is associated with the birth weight of newborns. Hence, Vitamin D levels during gestation may affect fetal growth and development

Correlation between maternal and neonatal blood vitamin D levels and its effect on the newborn anthropometry

Background: Vitamin D deficiency during pregnancy has important implications for the new born and infant. In India, there were few data about the prevalence of hypovitaminosis D in pregnancy and in new-born and its correlation. Hence, this study aim was to determine vitamin D deficiency in pregnant women and their new-borns and to assess the correlation between maternal and new born serum levels of the vitamin D3 and also to study its effect on the newborn anthropometry.Methods: This cross-sectional, hospital based study was conducted with 54 consecutive women in labour presenting with a singleton term pregnancy at a large tertiary centre in Tamilnadu, India. Data were recorded on a special form and 25-hydroxy vitamin D3 was assessed before delivery in mothers and after delivery in their infant’s umbilical cord blood.Results: The mean vitamin D levels were 20.21 ng/mL for the participants and 20.07 ng/mL for the newborns. Vitamin D severe deficiency was noted in 3 (6%), deficiency 31 (57%), insufficiency 12 (22%) and sufficiency in 8 (15%) of 54 participants whereas severe deficiency, insufficiency and sufficiency respectively, were noted in 4 (7%), 30 (56%), 11 (20%) and 9 (17%) of the newborns. Maternal vitamin D level significantly associated with socioeconomic status, term of delivery and type of delivery respectively were p=0.000, p=0.000 and p=0.038 whereas age (p=0.081), BMI (p=0.085) and parity (p=0.038) was not significant at p <0.05 level by chi-square analysis. Similarly, the new born characteristics of birth weight (p=0.000), birth length (p=0.016), head and chest circumference (p=0.001, p=0.000) exists significant association at p<0.05 level by analysis of variance (ANOVA) method and also there was a strong positive correlation between the vitamin D levels in maternal and cord blood (r=0.898; P <0.05).Conclusions: There was a high prevalence of vitamin D deficiency in the south Indian pregnant women and their newborns. Vitamin D deficiency increased the risk of low birth weight neonate and it also had an effect on the length of the baby, head circumference and chest circumference. This public health problem needs urgent attention

Sub-chronic toxicological evaluation of cleistanthin A and cleistanthin B from the leaves of Cleistanthus collinus (Roxb.)

AbstractObjectiveTo investigate the toxicological effects of cleistanthin A and cleistanthin B using sub-chronic toxicity testing in rodents.MethodCleistanthins A and B were isolated from the leaves of Cleistanthus collinus. Both the compounds were administered orally for 90 days at the concentration of 12.5, 25 and 50 mg/kg, and the effects on blood pressure, biochemical parameters and histology were assessed. The dose for sub-chronic toxicology was determined by fixed dose method according to OECD guidelines.ResultSub-chronic toxicity study of cleistanthins A and B spanning over 90 days at the dose levels of 12.5, 25 and 50 mg/kg (once daily, per oral) revealed a significant dose dependant toxic effect in lungs. The compounds did not have any effect on the growth of the rats. The food and water intake of the animals were also not affected by both cleistanthins A and B. Both the compounds did not have any significant effect on liver and renal markers. The histopathological analysis of both cleistanthins A and B showed dose dependent morphological changes in the brain, heart, lung, liver and kidney. When compared to cleistanthin A, cleistanthin B had more toxic effect in Wistar rats. Both the compounds have produced a dose dependent increase of corpora amylacea in brain and induced acute tubular necrosis in kidneys. In addition, cleistanthin B caused spotty necrosis of liver in higher doses.ConclusionThe present study concludes that both cleistanthin A and cleistanthin B exert severe toxic effects on lungs, brain, liver, heart and kidneys. They do not cause any significant pathological change in the reproductive system; neither do they induce neurodegenerative changes in brain. When compared to cleistanthin A, cleistanthin B is more toxic in rats

Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling

<p>Abstract</p> <p>Background</p> <p>Plant Growth Promoting Rhizobacteria (PGPR), <it>Pseudomonas fluorescens </it>strain KH-1 was found to exhibit plant growth promotional activity in rice under both <it>in-vitro </it>and <it>in-vivo </it>conditions. But the mechanism underlying such promotional activity of <it>P. fluorescens </it>is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to <it>P. fluorescens </it>treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry.</p> <p>Results</p> <p>Priming of <it>P. fluorescens</it>, 23 different proteins found to be differentially expressed in rice leaf sheaths and MS analysis revealed the differential expression of some important proteins namely putative p23 co-chaperone, Thioredoxin h- rice, Ribulose-bisphosphate carboxylase large chain precursor, Nucleotide diPhosphate kinase, Proteosome sub unit protein and putative glutathione S-transferase protein.</p> <p>Conclusion</p> <p>Functional analyses of the differential proteins were reported to be directly or indirectly involved in growth promotion in plants. Thus, this study confirms the primary role of PGPR strain KH-1 in rice plant growth promotion.</p

Evaluation of in vivo antitumor activity of cleistanthin B in Swiss albino mice

To evaluate the in vivo antitumor activity of cleistanthin B in Ehrlich's ascites carcinoma (EAC) and Dalton's ascites lymphoma (DAL) cell lines induced malignant ascites mouse models and DAL cell line induced solid tumor mouse model. All animals were injected with 2 × 106 EAC/DAL cells i.p./s.c. to induce malignant ascites and solid tumor and treated with 5-fluorouracil (5-FU) 20 mg/kg or cleistanthin B for 10 days. Cleistanthin B was given at three doses viz. 25, 50 and 100 mg/kg. The percentage increase in life span and the overall survival in malignant ascites animals and the tumor volume in solid tumor animals were measured. The haematological parameters were assessed in all animals before and 2 weeks after the treatment. Cleistanthin B 50 mg/kg and 5-FU significantly prolonged the life span (>25%) of malignant ascites tumor bearing animals. The overall survival was significantly improved by both. Only cleistanthin B 50 mg/kg significantly reduced the elevated WBC counts in EAC tumor bearing animals. Both 5-FU and cleistanthin B 50 mg/kg reversed the malignancy induced increase in neutrophils and platelet counts and decrease in lymphocyte counts but not to the normal range. Only 5-FU significantly reduced the solid tumor volume. None of the three doses of cleistanthin B was effective against the solid tumor. Cleistanthin B has antitumor activity against EAC and DAL tumor mice but it is not as effective as 5-FU. At 50 mg/kg dose cleistanthin B exerts significant antitumor activity compared to 25 and 100 mg/kg dose. Its effect on WBC count is higher and advantageous when compared to 5-FU. But cleistanthin B in the doses used is not effective against solid tumor

Association between placental vitamin D receptor expression and cord blood vitamin D level and its effect on the birth weight of newborns

Background: Vitamin D deficiency during pregnancy affects fetal growth and development. The present study explored the association between vitamin D levels in cord blood and placental vitamin D receptor expression.Methods: A total of 54 subjects with live-born singleton deliveries were included in SRM Medical College Hospital and Research Centre, Kattankulathur, India. Cord blood and placenta was obtained at the time of delivery.      Results: The mean newborn vitamin D level was 20.07 ng/mL (13-27.50). The mean of relative vitamin D receptor (VDR) expression was 1.45Ct (range 0.10-2.6). Mean comparison of cord serum 25(OH) D3 and VDR revealed significant at p&lt;0.05 by student’s t-test. The newborn anthropometric measurement of head circumference 31.44±3.99 cm, Chest circumference 28.88±4.30 cm, Body length 45.52±8.05 cm, Birth weight 2660.61±353.27 g had significant association with mean VDR expression at p &lt;0.05 level by student t-test analysis. Further, Pearson correlation coefficient also revealed strong positive correlation of placental VDR and cord serum vitamin D (r=0.853, p &lt;0.05).Conclusions: The present study indicates that there was a high prevalence of vitamin D deficiency in the south Indian newborns and the study suggested that VDR had crucial role in maternal to fetal nutrient transfer mechanism. Vitamin D status in cord blood is associated with the birth weight of newborns. Hence, Vitamin D levels during gestation may affect fetal growth and development

Correlation between maternal and neonatal blood vitamin D levels and its effect on the newborn anthropometry

Background: Vitamin D deficiency during pregnancy has important implications for the new born and infant. In India, there were few data about the prevalence of hypovitaminosis D in pregnancy and in new-born and its correlation. Hence, this study aim was to determine vitamin D deficiency in pregnant women and their new-borns and to assess the correlation between maternal and new born serum levels of the vitamin D3 and also to study its effect on the newborn anthropometry.Methods: This cross-sectional, hospital based study was conducted with 54 consecutive women in labour presenting with a singleton term pregnancy at a large tertiary centre in Tamilnadu, India. Data were recorded on a special form and 25-hydroxy vitamin D3 was assessed before delivery in mothers and after delivery in their infant’s umbilical cord blood.Results: The mean vitamin D levels were 20.21 ng/mL for the participants and 20.07 ng/mL for the newborns. Vitamin D severe deficiency was noted in 3 (6%), deficiency 31 (57%), insufficiency 12 (22%) and sufficiency in 8 (15%) of 54 participants whereas severe deficiency, insufficiency and sufficiency respectively, were noted in 4 (7%), 30 (56%), 11 (20%) and 9 (17%) of the newborns. Maternal vitamin D level significantly associated with socioeconomic status, term of delivery and type of delivery respectively were p=0.000, p=0.000 and p=0.038 whereas age (p=0.081), BMI (p=0.085) and parity (p=0.038) was not significant at p &lt;0.05 level by chi-square analysis. Similarly, the new born characteristics of birth weight (p=0.000), birth length (p=0.016), head and chest circumference (p=0.001, p=0.000) exists significant association at p&lt;0.05 level by analysis of variance (ANOVA) method and also there was a strong positive correlation between the vitamin D levels in maternal and cord blood (r=0.898; P &lt;0.05).Conclusions: There was a high prevalence of vitamin D deficiency in the south Indian pregnant women and their newborns. Vitamin D deficiency increased the risk of low birth weight neonate and it also had an effect on the length of the baby, head circumference and chest circumference. This public health problem needs urgent attention