An innovative and cost-effective way to estimate alkaline phosphatase activity in in vitro cellular model systems

Alkaline phosphatase is an enzyme that converts para-nitrophenyl phosphate to para-nitrophenol (yellow coloured) in 2-amino, 2-methyl, 1-propanol buffer at pH 10.5. However, when this protocol is applied to the in vitro cellular model systems to estimate alkaline phosphatase activity, it tends to generate clumps of genomic DNA, leading to inaccurate pipetting for protein estimation. The aim of the study was to introduce minor modifications in the existing protocol to make it simple, cost-effective, with minimal labor-intensive procedures while estimating alkaline phosphatase activity in cellular model systems. The genomic DNA clumps were dissolved by depurination (adding 0.2 N HCl) and fragmentation (adding 0.2 N NaOH) during enzyme estimation. Moreover, these minor modifications have been standardized and optimized extensively by using serum samples (rich source of alkaline phosphatase), hFOB/ER9 (human Fetal osteoblastic cell) and HepG2 cells. Our results suggest that the modification incorporated in previously published method was robust enough to estimate ALP activity and protein concentration accurately. There was no significant variation in ALP activity estimated after modification (P > 0.05). This innovative approach could be beneficial for a researcher by providing an easy, cost effective and less labor-intensive solution for estimation of enzymatic activity in cellular model systems.     

Gain-of-function p53 protein transferred via small extracellular vesicles promotes conversion of fibroblasts to a cancer-associated phenotype

1. Download : Download high-res image (201KB)
2. Download : Download full-size image

     

Phage-typing ofSalmonella weltevreden based on lysogeny I. The phage-typing system

All of the 1070 strains of Salmonella weltevreden tested were lysogenic, temperate phage being demonstrable in 83.9% of the strains in broth culture, 6.4% in mixed culture and the remaining 9.7% after induction by UV-irradiation and MC treatment. A phage-typing scheme for Salmonella weltevreden based on the host range of temperate phages was developed. The strains could be classified into eight groups by means of the host range of their temperate phages on six indicator strains and a ninth group, the symbiotic phages of which failed to lyse any of the indicator strains.     

Effect of denervation on loss of ions in chick skeletal muscle

Leaching behaviour of chick gastrocnemius muscle has been studied with respect to the loss of Na+, K+ and Ca2+ ions in a donor-solvent (muscle-water) system under normal as well as denervated conditions. An attempt has been made to explain the alterations in the rate of electrolytic loss in terms of membrane dysfunctions induced as a result of loss of neural control.     

Chromosomal polymorphism in Cassida syrtica Boh. (Coleoptera: Chrysomelidae)

Intraspecific polymorphism with regard to autosomes has been observed in Cassida syrtica, where an additional large autosome (marker chromosome) has been encountered in the diploid as well as the haploid set of chromosomes. The diploid number for the normal type is 18 (n = 8 + Xyp). Its polymorphic form reveals a diploid set of ninetten chromosomes with an exceptional karyotype (n = 8 + 1 Mt + Xyp).     

The effect of opioids and their antagonists on the nocifensive response of Caenorhabditis elegans to noxious thermal stimuli

Opiates modulate nociception in vertebrates. This has also been demonstrated in a number of invertebrate models. Herein, the effect of the opiate morphine and opioid neuropeptides Endomorphin 1 and 2 on the thermal avoidance (Tav) behavior of Caenorhabditis elegans is explored. Adult wild-type C. elegans N2 were collected from NGM plates using M9 buffer and exposed to morphine and endomorphine 1 and 2 in concentrations between 10^?8 and 10^?4 M (2.5 pmol/mg to 25 nmol/mg) for 30 min and tested for Tav. The opioid receptor antagonists Naloxone and CTOP were tested in combination with the drugs. Forty-seven percentage of the morphine exposed worms exhibited a class I response versus 76% of the control group (P < 0.001). Endomorphin 1 and 2 also caused a statistically significant reduction in class I responses, 36 and 39%, respectively. These effects were reversed with Naloxone and CTOP. Thermonocifensive behavior in C. elegans is modulated by opioids.