Secretory TAT-peptide-mediated protein transduction of LIF receptor α-chain distal cytoplasmic motifs into human myeloid HL-60 cells

The distal cytoplasmic motifs of leukemia inhibitory factor receptor α-chain (LIFRα-CT3) can independently induce intracellular myeloid differentiation in acute myeloid leukemia (AML) cells by gene transfection; however, there are significant limitations in the potential clinical use of these motifs due to liposome-derived genetic modifications. To produce a potentially therapeutic LIFRα-CT3 with cell-permeable activity, we constructed a eukaryotic expression pcDNA3.0-TAT-CT3-cMyc plasmid with a signal peptide (ss) inserted into the N-terminal that codes for an ss-TAT-CT3-cMyc fusion protein. The stable transfection of Chinese hamster ovary (CHO) cells via this vector and subsequent selection by Geneticin resulted in cell lines that express and secrete TAT-CT3-cMyc. The spent medium of pcDNA3.0-TAT-CT3-cMyc-transfected CHO cells could be purified using a cMyc-epitope-tag agarose affinity chromatography column and could be detected via SDS-PAGE, with antibodies against cMyc-tag. The direct administration of TAT-CT3-cMyc to HL-60 cell culture media caused the enrichment of CT3-cMyc in the cytoplasm and nucleus within 30 min and led to a significant reduction of viable cells (P < 0.05) 8 h after exposure. The advantages of using this mammalian expression system include the ease of generating TAT fusion proteins that are adequately transcripted and the potential for a sustained production of such proteins in vitro for future AML therapy.

Effect of gala apples (Malus domestica Borkh) on lipidemia of hyperlipidemic rats

A healthy life style and a balanced diet, associated with a high fruit and vegetable intake, are linked to good health and the prevention of diseases. Apples contain bioactive compounds that help in the prevention and control of hyperlipidemia. One of the Public Health concerns in Brazil is to reduce cardiovascular diseases. Therefore, the objective of this work was to analyze the chemical composition of Gala apples and to study the effect of their consumption on weight gain, food intake, serum levels of total cholesterol, HDL-C, LDL-C, triglycerides, hepatic cholesterol and fecal cholesterol in male albino Wistar rats fed a hypercholesterolemic diet. Six animals were utilized for each treatment (control, 5, 15 and 25% apple diet), during 30 and 60 days. This study showed that one apple (200 g) can provide 14.5% of recommended total fiber and 55% of recommended vitamin C, besides supplying considerable quantities of phenolic compounds (0.38 g.100 g-1) and tannins (0.16 g.100 g-1). All animals showed a non-significant reduction in their weight gain and food intake with an increase in the concentration of apple in the diets. At the end of 30 days, all of the diets provided a significant reduction in the levels of triglycerides compared to the control group. The 15 and 25% apple diets showed significant reductions in the serum levels of total cholesterol and LDL-C and an increase in the level of fecal cholesterol in relation to the control group. The 25% apple diet provided a significant reduction in the hepatic cholesterol levels compared to the control group. After 60 days, the serum levels of total cholesterol, LDL-C, HDL-C and triglycerides in rats fed with 5, 15 and 25% apple diets were similar to the control group. This probably happened due to a revertion of the process. These results show the importance of Gala apples in the control of hyperlipidemia in rats. A diet rich in vegetables and fruits, including apples, associated to a healthy life style, over time, could prevent or reduce the risk of heart disease.

Effects of defatted amaranth (Amaranthus caudatus L.) snacks on lipid metabolism of patients with moderate hypercholesterolemia

We evaluated the effects of defatted amaranth (Amaranthus caudatus L.) snacks on plasma lipids in moderate hypercholesterolemic patients. Twenty-two subjects [30-65 years old), 11 males, with total cholesterol (TC) > 240 mg.dL-1, low-density cholesterol (LDL-c) 160-190 mg.dL-1 and plasma triglycerides (TG) < 400 mg.dL-1] were randomized in a double blind clinical trial to receive an amaranth snack (50 g/day) or equivalent corn snack (placebo) for 2 months. There were no differences between amaranth and placebo on TC and LDL-c, and TG respectively: -8.4 and -5.7% (p = 0.17); -12.3 and -9.7% (p = 0.41) and -0.6 and -7.3% (p = 0.47). However, amaranth snacks significantly reduced high-density cholesterol (HDL-c): -15.2 vs. -4% (p = 0.03). In conclusion, the intake of 50 g of extruded amaranth daily during 60 days did not significantly reduce LDL-c in moderate hypercholesterolemic subjects; furthermore there was a significant reduction in HDL-c. Studies with greater number of subjects and greater quantity of this food are necessary to test the effects of amaranth on lipid metabolism in humans.