Archive for December, 2014

Fibronectin Expression and Gene Transcription in Fibroblasts

December 12th, 2014 No comments

Like other fibroblasts, cultured NIH-3T3 fibroblasts produce fibronectin in a constitutive fashion, and we have previously demonstrated that nicotine greatly enhances fibronectin expression in these cells. To examine the effects of FP on fibronectin protein expression, NIH-3T3 fibroblasts were pretreated in the presence or absence of FP for 1 h, cultured with or without nicotine (50 pg/mL) for 48 h, followed by harvesting and processing for western blotting.

Fibronectin Expression

Canadian Pharmacy NORX

FP pretreatment inhibited the production of fibronectin protein in both untreated and nicotine-stimulated fibroblasts. In addition, nicotine also stimulated an increase in fibronectin mRNA of > 2.5-fold (p < 0.03) as compared to untreated control cells. Preincubation of fibroblasts for 1 h with FP was associated with a significant reduction (p < 0.04) in fibronectin mRNA accumulation in nicotine-treated fibroblasts as determined by real-time PCR.

The observations described above suggested that FP might exert its effect by inhibiting fibronectin gene transcription. To test this, NIH-3T3 fibroblasts were stably transfected with pFN(1.2kb)LUC, a DNA construct containing the human fibronectin promoter fused to a luciferase reporter gene.

As expected, the transfected cells showed constitutive fibronectin gene transcription as determined by measuring luciferase activity. The transcription of the fibronectin gene was enhanced by treatment with nicotine alone . Of note, the exposure of fibroblasts to FP inhibited the transcription of the fibronectin gene at baseline and after nicotine stimulation .

The effect of FP pretreatment on nicotine-induced fibronectin gene transcription was exerted in a dose-dependent manner with maximum activity between 1 and 5 pmol/L FP . The same effect was observed in primary mouse lung fibroblasts containing pFN(1.2 kb)LUC, where FP pretreatment significantly inhibited transcription of the fibronectin gene after nicotine stimulation.

We and others have shown that fibronectin gene transcription is dependent on phosphorylation and DNA binding by the transcription factor CREB. Consequently, we postulated that FP might affect fibronectin expression by inhibiting CREB. Consistent with this idea, we found that nicotine-treated fibroblasts show increased CREB phosphorylation when compared to untreated fibroblasts. FP pretreatment diminished phosphorylated CREB after nicotine stimulation, whereas the amount of total CREB protein remained relatively unchanged.

Treatment Through Gymnastic Exercises

December 10th, 2014 No comments

Left arm forward and slightly tilted, palm down. The right hand moves along the left arm. Start from the shoulder and go downward, while you are singing the exercise with the letter “Y” (the Bulgarian letter “Y” is pronounced [u] -transl. note). Then the hand goes down to the wrist, while you are singing the exercise with the letter “A” (the Bulgarian letter “A” is pronounced [a] – transl. note). Through Gymnastic Exercises

Finally the hand goes up from the wrist to the elbow, while you are singing the letter “H” (the Bulgarian letter “H” is pronounced [i] – transl. note). The distance from the shoulder to the elbow is the material world, the physical world of the stomach. The zone from the elbow to the wrist is the spiritual world – the heart, and the wrist is the mental world – the mind. Then put your right arm forward, and the left hand is to be moved along it.

Do this exercise for a week every morning and evening for ten times. This will improve the state of the stomach, the chest and the brain. You will treat yourself. One, who wants to be healthy, shall sing. If one is sad or does not feel well, he shall sing, too. Sing and praise God in your soul.

Right leg forward, easily bend the body to the earth, while imaging that you are lifting an enormous load. Do this exercise, gradually increasing the load until you reach 50 kg. By doing the exercise regularly, you will notice that your muscle strength has increased.

If you are able to lift 50 kg mentally and you will be also able to really lift it. Do this exercise when you do not feel well to see what powers you have.

In order your eyes not to weaken, you shall practice. There are exercises, which stir all eye muscles. If not exercised, eyes gradually weaken and lose their ability to see. When you go on an excursion, you want to reach the peak as soon as possible. This is not right. Go slowly and look up, left, right. Once the sun is up, stop for a while, to see how it rises and continue your way. If you are in a hurry, you will soon get old.

One of the reasons for weakening of the eyes is that they do not move. You move your neck when you want to see something and do not move your eyes. I would recommend an exercise that you can do in the morning and whenever you want during the day. Close your eyes and move them, look, as much as it is possible, up, then, as much as it is possible, down, as much as it is possible to the right, and as much as it is possible to the left.

Repeat this several times very slowly. Then, during the day, when you want to see something, do not twist your neck. The neck shall be still, and the eyes are to be moved -this will become the natural way of looking at things for you. This is the physical side. Inside is the following: if you make a mistake and do not fix it, then if you make a second mistake and do not fix it, third one, fourth one, you get used to that and your sight weakens. Another thing: You love someone and excuse his mistakes or you see them as too small ones. You do not love him, do not excuse his mistakes and you see them as too big. Third: eyes weaken as a result of worries, concerns, internal disharmonious states, or of dust.

Cold feet are the first condition of neurasthenia. It is good for neurotics and those, who often lose their temper, to remove their shoes from time to time and walk barefoot on sand or soft grass. In this way feet are in direct contact with the vital powers of the earth and tone. In the morning exercises, while lifting your arms up, you take in the solar energy and through legs – the earth energy. Hence, in the morning, you shall take in solar energies, and in the evening you shall do exercises to get rid of the energy, collected during the day. In the morning you will take in from the Sun, and in the evening you will give the excessive energy to the Earth.