Staurosporine augments EGF-mediated EMT in PMC42-LA cells through actin depolymerisation, focal contact size reduction and Snail1 induction – A model for cross-modulation

<p>Abstract</p> <p>Background</p> <p>A feature of epithelial to mesenchymal transition (EMT) relevant to tumour dissemination is the reorganization of actin cytoskeleton/focal contacts, influencing cellular ECM adherence and motility. This is coupled with the transcriptional repression of E-cadherin, often mediated by Snail1, Snail2 and Zeb1/δEF1. These genes, overexpressed in breast carcinomas, are known targets of growth factor-initiated pathways, however it is less clear how alterations in ECM attachment cross-modulate to regulate these pathways. EGF induces EMT in the breast cancer cell line PMC42-LA and the kinase inhibitor staurosporine (ST) induces EMT in embryonic neural epithelial cells, with F-actin de-bundling and disruption of cell-cell adhesion, via inhibition of aPKC.</p> <p>Methods</p> <p>PMC42-LA cells were treated for 72 h with 10 ng/ml EGF, 40 nM ST, or both, and assessed for expression of E-cadherin repressor genes (Snail1, Snail2, Zeb1/δEF1) and EMT-related genes by QRT-PCR, multiplex tandem PCR (MT-PCR) and immunofluorescence +/- cycloheximide. Actin and focal contacts (paxillin) were visualized by confocal microscopy. A public database of human breast cancers was assessed for expression of Snail1 and Snail2 in relation to outcome.</p> <p>Results</p> <p>When PMC42-LA were treated with EGF, Snail2 was the principal E-cadherin repressor induced. With ST or ST+EGF this shifted to Snail1, with more extreme EMT and Zeb1/δEF1 induction seen with ST+EGF. ST reduced stress fibres and focal contact size rapidly and independently of gene transcription. Gene expression analysis by MT-PCR indicated that ST repressed many genes which were induced by EGF (EGFR, CAV1, CTGF, CYR61, CD44, S100A4) and induced genes which alter the actin cytoskeleton (NLF1, NLF2, EPHB4). Examination of the public database of breast cancers revealed tumours exhibiting higher Snail1 expression have an increased risk of disease-recurrence. This was not seen for Snail2, and Zeb1/δEF1 showed a reverse correlation with lower expression values being predictive of increased risk.</p> <p>Conclusion</p> <p>ST in combination with EGF directed a greater EMT via actin depolymerisation and focal contact size reduction, resulting in a loosening of cell-ECM attachment along with Snail1-Zeb1/δEF1 induction. This appeared fundamentally different to the EGF-induced EMT, highlighting the multiple pathways which can regulate EMT. Our findings add support for a functional role for Snail1 in invasive breast cancer.</p

PK/Statistical model in depression hypertension.

Background: Depression hypertension is a co-morbid disease and needs treatment with more than one drug. Material & Methods: 25 patients each from depression hypertension and hypertension were included in the study. In order to evaluate the PK/PD statistical model, single dose 100mg Amitriptyline was administered to the depression hypertension and hypertension subjects. PK parameters of Amitriptyline and its metabolite, nortriptyline was determined. The concentration of Amitriptyline and Nortriptyline was estimated by HPLC, which is the versatile, accurate and sensitive method. Plasma samples at 0, 1, 2, 4, 8, 12, 24 hours were drawn from each subject along with the Pharmacodynamic changes like blood pressure, heart rate and respiratory rate. After a wash-out period of 7 days, Amlodipine 10mg was administered to each group when 10-15mmHg diastolic blood pressure was decreased and then the Amitriptyline 100mg wad again administered. PK and PD parameters were again estimated. The groups were as follows: T1 and C1 of 23 patients each. These 2 groups were sub-divided into four groups i. E. (1) T1a (Depressive hypertension before Amlodipine), (2) T1b (Depressive hypertensive after Amlodipine), C1a (Hypertensive before amlodipine) and C1b (Hypertensive after Amlodipine). Results: Pharmacokinetic parameters of Amitriptyline like T½, Cmax, Tmax, AUC, Vd, clearance of these two groups were compared. T½ (hours) of C12, C1b were 20.213±4.51 and 23.78±3.99 having P value <.05, T ½ of T1a, T1b were 19.805±5.66 and 20.327±4.27 having p. Value of 0.752. Cmax (ng/ml) of C1a, C1b were 31.88±13.54 and 32.45±13.14 having p. Value of 0.649, Cmax of T1a and T1b were 30.90±11.8 and 31.08±12.0 having p. value of 0.584. AUC (ng/ml into hour) of C1a and C1b were 186.09±44.18 and 203.88±43.39 having p. value <0.05, AUC of T1a and T1b¬ were 182.80±39.6 and 210.57±48.86 having p. value <0.05. Volume of distribution (Vd l/kg) of C1a and C1b were 15.18±2.43 and 18.78±2.50, Vd of T1a and T1b were 14.61±3.43 and 16.56±2.54 having p. value <0.05. Clearance (l/hr) of C1a and C1b were 9.55±2.6 and 8.59±2.32 having p. value of 0.129, clearance of T1a and T1b were 9.59±2.32 and 8.59±2.32 having p. value <0.05. Conclusions: There is a significant change of PK parameters in the group C1a and C1b in T½, AUC and Vd. In the group T1a and T1b, there is also a significant change in AUC, Vd and Clearance as well. So by changing the blood flow, there is a significant change of PK parameters in the object drug Amitriptyline

Ecological Risk Assessment of Metal Pollution along Greater Cairo Sector of the River Nile, Egypt, Using Nile Tilapia, Oreochromis niloticus, as Bioindicator

The present work aims to evaluate seasonal metal pollution along Greater Cairo sector of the River Nile, Egypt, using wild Nile tilapia, Oreochromis niloticus, as bioindicator and to conduct a risk assessment for human consumers. Greater Cairo is the largest populated area along the whole course of River Nile with a wide range of anthropogenic activities. Effects of metal pollution on fish body indices were studied using condition factor (CF) and scaled mass index (SMI). Metal pollution index (MPI) showed that the total metal load in fish organs followed the follwoing order: kidney > liver > gill > muscle which gives a better idea about the target organs for metal accumulation. Metal concentrations in fish muscle (edible tissue) showed the following arrangement: Fe > Zn > Cu > Mn > Pb > Cd. Metal’s bioaccumulation factor (BAF) in fish muscle showed the following arrangement: Zn > Cu > Fe > Mn > Cd and Pb. The hazard index (HI) as an indicator of human health risks associated with fish consumption showed that adverse health effects are not expected to occur in most cases. However, the metals’ cumulative risk effects gave an alarming sign specifically at high fish consumption rates

Impact of nanosilver-profenofos on cotton leafworm, Spodoptera littoralis (Boisd.) larvae

Abstract Background The cotton leafworm, Spodoptera littoralis (Boisd.), is one of the most dangerous pests in Egypt and Africa, causing significant economic loss of cotton crop yield. The frequent use of insecticides to control this insect has led to the development of a generation’s resistance to them. The need of a new, safe control method and effective insecticides has become necessary after the recent increases in environmental pollution and insect resistance. This study is devoted to developing a novel synthetic scheme to produce a pesticide nanocomposite of very high efficiency compared to the originals. The nature of its chemical binding has been investigated via Fourier transform-infrared (FT-IR) and transmission electron microscopy (TEM) techniques. Results In this work, the method is based on using silver nanoparticles (AgNPS) as a pesticide carrier by loading the organophosphorus pesticide profenofos on to their surface. The profenofos, AgNPS alone and nanocomposite profenofos (AgNPS@P) have been tested against second- and fourth-instar larvae of laboratory and field cotton leafworm. Our findings indicate that the AgNPS@P is more effective on cotton leafworm larvae than each of profenofos and nanosilver alone. The activity of AgNPS@P (LC50 = 0.94 and 5.15 ppm) was increased to 85 and 69 times more than that of profenofos (LC50 = 79.52 and 356.97 ppm) against second- and fourth-instar larvae of field cotton leafworm. Conclusions This method may be successful for reducing environmental pollution and the resistance of this pest to many pesticides