The E1 Copper Binding Domain of Full-Length Amyloid Precursor Protein Promotes Epithelial to Mesenchymal Transition in DU145 Cells in an Isoform-Specific Manner
Epithelial to mesenchymal transition (EMT) confers migratory and dynamic properties on cells and, as such, plays a pivotal role in the development of metastatic, castration resistant prostate cancer. The amyloid precursor protein (APP), although most closely associated with the neurodegenerative condition Alzheimer's disease, has also been linked to the pathogenesis and prognosis of several cancers including prostate cancer.
Aims: To investigate whether over-expression of APP could promote EMT in prostate cancer (PCa) DU145 cells and to determine the molecular prerequisites for this effect.
Methodology: A range of APP molecular constructs were stably expressed in DU145 cells and their effects on EMT were monitored by morphological analysis and by immunoblotting for the EMT marker proteins, E-cadherin and vimentin.
Results: Our results show that the full-length 695 amino acid isoform (APP695), but not APP751 or APP770, promoted EMT via a mechanism requiring an intact extracellular E1 copper binding domain and tyrosine687 within the cytosolic domain of the protein.
Conclusion: Targeting the expression of APP695 or the E1 copper binding domain of the protein may, therefore, contribute to therapeutic strategies for the delay or prevention of prostate cancer metastasis.
Evaluation of the Effects of Methanolic Extract of Alligator Pepper (Aframomum melegueta) on Serum creatinine, Blood Urea Nitrogen and Electrolytes in Wistar Albino Rats
The effect of methanolic extract of Alligator pepper (Aframomum melegueta) (AME) on the serum creatinine, blood urea nitrogen (BUN) and electrolytes were investigated in Wistar albino rats. The extraction was done by cold maceration of pulverized seeds in absolute methanol for 48 hours. The extract was filtered and concentrated in vacuo in a rotary evaporator at 40ºC. Twenty four male albino Wistar rats were randomly assigned into four groups of six animals each. Group A served as the control and received 10 ml/kg of 5 % Tween 20 while Group B-D received 100, 200 and 400 mg/kg respectively, of the AME for 21 days. Twenty four hours later, blood was collected from the rats through direct cardiac puncture. The separated serum was used to determine the serum creatinine, BUN and electrolyte assay. The extract produced no significant (p > 0.05) difference in mean serum creatinine, BUN and electrolytes level in the treated rats when compared to the control group rats. In conclusion the extract did not exhibit any adverse effect on the kidney at the doses used and for the period of experiment.
Prognostic Value of Leptin Gene Polymorphisms in Type 2 Diabetes Mellitus Patients in Egypt
Aim: The work wasto study polymorphisms in the LEP gene in type 2 diabetics in Minia, Egypt and determined the relationship between the leptin and c-peptide levels in different genotypes and insulin resistance in obese patients. The study also has evaluated the role of leptin gene polymorphism in prediction of diabetes mellitus prognosis and its prevention.
Study Design: Investigative.
Place and Duration of Study: Samples were analyzed at Biochemistry Department, Faculty of Medicine- Minia University, Egypt, between August 2012 and April 2014.
Methodology: This study was performed in 80 patients with type 2 diabetes mellitus (54 men and 26 women) and 15 normal controls (12 men and 3 women). In our thesis we measured HbAc, fasting blood glucose, leptin hormone and c-peptide. DNA extracted and the human leptin gene (for product 242 bp) was amplified by PCR, Restriction analysis of the PCR products was performed with restriction enzyme HhaI and genotyped at the restriction site located -2549 bp from the transcription initiation site of leptin gene. Presence of allele (C at -2549 bp) and absence of allele (A at the same position) were identified through the GCGC sequence.
Results: Our study showed C-2549 A variant is associated with the fasting leptin levels, the results which are in agreement with previous studies. The polymorphism in leptin gene has an effect on the level of plasma leptin in different genotypes, and individuals with AA genotype have the lower plasma levels of leptin and also c-peptide than AC and CC variants.
Conclusion: Our study reveals that both diabetic patients and their non-diabetic relatives have different basal leptin and c-peptide specific to different leptin genotypes. This suggests that the association between leptin and insulin in members of diabetic families may be controlled by inheritance.
Antiplasmodial Activity of Ethanolic Leaf Extracts of Spilanthes uliginosa, Ocimum basilicum
(Sweet Basil), Hyptis spicigera and Cymbopogon citratus on Mice Exposed to Plasmodium berghei
In Africa and elsewhere, medicinal plants including Spilanthes uliginosa, Ocimum basilicum, Hyptis spicigera and Cymbopogon citratus are still widely used in the treatment of malaria and other ailments. The aim of the present study was to investigate in vivo antiplasmodial effect of ethanolic leaf extract of these plants in mice. Oral acute toxicity of the extracts was evaluated in mice using modified Lorke’s method and their in vivo anti-plasmodial effect against early infection, curative effect against established infection and prophylactic effect against residual infection were studied using total WBC count in chloroquine-sensitive Plasmodium berghei NK 65-infected mice. The oral median lethal dose of the extract in mice was determined to be greater than 2000 mg kg-1 body weight. The results indicated a significant (P<0.05) daily increase in the level of parasitaemia in the parasitized untreated groups and a significant (P<0.05) dose dependent decrease in the level of parasitaemia in the parasitized groups treated with varying doses of the various medicinal plants and the standard drug. Overall, the dose dependent effects were in the order of: 5mg/kg body weight of chloroquine > 800 mg/kg > 400 mg/kg > 200 mg/kg body weight of the plant extracts with the efficacy of the plants in the order of: H. Spicigera > O. basilicum > C. citratus > S. uliginosa (Sw) with minor variations. The implications of these results is that Spilanthes uliginosa, Ocimum basilicum, Hyptis spicigera and Cymbopogon citratus ethanolic leaf extracts posses potent antimalarial effects and may therefore serve as potential sources of safe, effective and affordable antimalarial drugs.
Clinical Evaluation and Serum Lipid Profile between Individuals with Acute Hepatitis C
Aim: Hepatitis C virus (HCV) exerts an intense impact on host lipid metabolism. It has been shown that the synthesis of cholesterol and fatty acids (FA) is directly affected in HCV patients but serum FA profile of acute HCV patients have not been directly quantified in humans.
Methodology: In present study the serum lipid and FA’s profile (free and total) of acute hepatitis C patients (n=50) is evaluated in comparison to healthy controls (n=50). The acute HCV patient’s were diagnosed by center of diseases control (CDC) criteria. Blood hematology, serum proteins, enzymes, waste metabolites and nutrition status were also assessed by standard methods.
Results: The acute HCV patients have significantly lower (P<0.05) lipid profile including triglycerides, cholesterol, HDL and LDL in relation to controls. Results of serum lipid FA’s (total and free form) reveal elevated level of saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA), with lower content of polyunsaturated fatty acid (PUFA) in acute hepatitis C patient as compared to controls. Among SFA, myristic and palmitic acid were increased and five unsaturated FA including nervonic, linoleic, α-linolenic, docosahexaenoic and arachidonic acid were reduced than control subjects. Significantly higher (P<0.05) alanine transaminase, alkaline phosphatase, direct bilirubin and globulin levels were found in acute HCV patients possibly due to viral infection.
Conclusion: This work makes available direct proof that lipogenesis is elevated in acute HCV patients while as evident from reduced serum PUFA and elevated SFA as well as MUFA. Hence acute HCV patients have low PUFA levels, which put forward importance of PUFA supplementation.