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His team teased apart the critical difference between normal and cancerous cells by using the fluorescence of natural NADH.
Combining the state-of-the-art spectroscopy and microscopy techniques, he and his colleagues were able to convert such fluorescence into an accurate measure of NADH concentration in live cells.
Heikal and graduate student Yu have found that the average concentration of NADH in breast cancer cells is about twice that in normal breast cells.
“If we are given two live cells, one normal and the other cancerous, we could differentiate between the two with confidence,” said Heikal.
“For the first time, we have been able to quantify the concentration of NADH in both live breast cells and breast cancer cells,” he added.
The research team also looked at the amounts of NADH in the cell that is free and how much is bound to other enzymes. The amounts were found to be different in normal and cancer cells.
“We realized that the fluorescence intensity not only depends upon the concentration of NADH but also on its structure — free or enzyme-bound — as well as its place inside the cell — in the cytoplasm (non-nucleus part of the cell) or in mitochondria,” said Heikal.
“Since a free NADH molecule would rotate — tumble — faster than enzyme-bound NADH, we were able to develop a technique called rotational diffusion imaging to establish a direct measure of the concentrations of free and enzyme-bound NADH throughout a living cell, whether in the cytosol (cell fluid) or the mitochondria,” he added.
Heikal and Yu confirmed that disruption of chemical reactions that produce ATP could lead to an increase in NADH by exposing normal breast cells to potassium cyanide, a known inhibitor of some of these critical mitochondrial enzymes.
They found that the NADH concentration in the normal cells increased when exposed to potassium cyanide. The relative amounts of NADH in the mitochondria also rose significantly.
While previous studies measured the amount of NADH in cells using conventional biochemical techniques that require destroying the cells, Heikal believes that measurements of dead cells may not be accurate or relevant for diagnostic or clinical use.
“The advantage of our non-destructive approach is that the NADH location in a cell relates to its function in cell survival,” he said.
“When you destroy the cell, you do not know where the NADH molecules existed inside the cell and what role they might have played in cell survival. For accurate diagnosis, you need to have the cellular context to better understand the problem,” he added.
He is of the opinion that the ability to accurately measure NADH levels in a cell without killing it could have potential implications for related research on human health and drug delivery.
“Our technique is not limited to detecting cancer. Other neurodegenerative diseases related to mitochondrial anomalies can also be detected with our method,” Heikal said.
“We can also use our approach to quantify the efficiency of a new drug on manipulating the activities of mitochondrial enzymes associated with energy production in cells,” he added.
The findings of the study have been published in the Journal of Photochemistry and Photobiology B: Biology. (ANI)
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The life of Marie Schlau, a German Jewish girl born in 1833 hides great unsolved mysteries: accidents, disappearances, enigmas, unknown diagnoses, disturbing murders, love, tenderness, greed, lies, death ... alternatively a different story unfolds every time and takes us closer to the present. Thus, there are two parallel stories unravelling, each in a different age and place, which surprisingly converge in a revelatory chapter.
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Currently, BabelFAmily is financing two promising research projects aimed at finding a cure for Friedreich's Ataxia. Whenever you make a donation to us or purchase a copy of "The legacy of Marie Schlau", this is where all funds raised will be devoted to:
1) Gene Therapy for Friedreich's Ataxia research project:
The project is the result of an initiative of Spanish people affected by this rare disease who are grouped in GENEFA in collaboration with the Spanish Federation of Ataxias and the BabelFAmily. The Friedreich’s Ataxia Research Alliance (FARA), one of the main patients’ associations in the United States now joins the endeavour.
2) Frataxin delivery research project:
The associations of patients and families Babel Family and the Asociación Granadina de la Ataxia de Friedreich (ASOGAF) channel 80,000 euros of their donations (50% from each organisation) into a new 18-month project at the Institute for Research in Biomedicine (IRB Barcelona). The project specifically aims to complete a step necessary in order to move towards a future frataxin replacement therapy for the brain, where the reduction of this protein causes the most damage in patients with Friedreich’s Ataxia.
The study is headed by Ernest Giralt, head of the Peptides and Proteins Lab, who has many years of experience and is a recognised expert in peptide chemistry and new systems of through which to delivery drugs to the brain, such as peptide shuttles—molecules that have the capacity to carry the drug across the barrier that surrounds and protects the brain. Since the lab started its relation with these patients’ associations in 2013*, it has been developing another two projects into Friedrich’s Ataxia.