Is It Alive or Dead?

How to Measure the Thermal Signatures of Single Cells

and Assess Their Biological Activity
To the ancients, probing the philosophical question of how to distinguish the living from the dead
centered on the "mystery of the vital heat." To modern microbiology, this question was always less
mysterious than it was annoying -- researchers have known that biological processes should
produce thermal signatures, even within single cells, but nobody ever knew how to measure them.
Now, a group of mechanical engineers from Pohang University of Science and Technology in Korea
have discovered a way to measure the "thermal conductivity" of three types of cells taken from
human and rat tissues and placed in individual micro-wells. They showed that they could detect
uniform heat signatures from the various cells and measured significant difference between dead
and living ones, suggesting a new way to probe cells for biological activity.
A lone cell is fantastically small, often only about 10 microns across (10 millionths of a meter), and
this size has thwarted thermodynamic measurements of single cells. Writing in the journal Applied
Physics Letters, a team led by Dongsik Kim and Jaesung Park describes how their novel nanoscale
biosensing technique can measure the thermal conductivity of a single cell.
"In the short-term, this biosensing technique can be used to measure cell viability," said Kim. "In the
long-term, we hope to refine it to develop a non-invasive, rapid means for early diagnosis of
diseases such as cancer based on differences in the thermal properties of cells."
While the fundamental heat signatures the researchers detected are not exactly what the ancient
philosophers imagined, measuring them may answer more mysteries than they could have dreamed.
The article, "Thermal conductivity of single biological cells and relation with cell viability" by Byoung
Kyoo Park, Namwoo Yi, Jaesung Park, and Dongsik Kim appears in the journal Applied Physics
Letters.
Reference:
http://www.sciencedaily.com/releases/2013/06/130628102927.htm

Mimicking Living Cells: Synthesizing Ribosomes

Synthetic biology researchers at Northwestern University, working with partners at Harvard Medical School, have for the first time
synthesized ribosomes -- cell structures responsible for generating all proteins and enzymes in our bodies -- from scratch in a test
tube.
Others have previously tried to synthesize ribosomes from their constituent parts, but the efforts have yielded poorly functional
ribosomes under conditions that do not replicate the environment of a living cell. In addition, attempts to combine ribosome synthesis
and assembly in a single process have failed for decades.
Michael C. Jewett, a synthetic biologist at Northwestern, George M. Church, a geneticist at Harvard Medical School, and colleagues
recently took another approach: they mimicked the natural synthesis of a ribosome, allowing natural enzymes of a cell to help facilitate
the human-made construction.
The technology could lead to the discovery of new antibiotics targeting ribosome assembly; an advanced understanding of how
ribosomes form and function; and the creation of tailor-made ribosomes to produce new proteins with exotic functions that would be
difficult, if not impossible, to make in living organisms.
"We can mimic nature and create ribosomes the way nature has evolved to do it, where all the processes are co-activated at the same
time," said Jewett, who led the research along with Church. "Our approach is a one-pot synthesis scheme in which we toss genes
encoding ribosomal RNA, natural ribosomal proteins, and additional enzymes of an E. coli cell together in a test tube, and this leads to
the construction of a ribosome."
Jewett is an assistant professor of chemical and biological engineering at Northwestern's McCormick School of Engineering and
Applied Science.
The in vitro construction of ribosomes, as demonstrated in this study, is of great interest to the synthetic biology field, which seeks to
transform the ability to engineer new or novel life forms and biocatalytic ensembles for useful purposes.
The findings of the four-year research project were published June 25 in the journal Molecular Systems Biology.
Comprising 57 parts -- three strands of ribonucleic acid (RNA) and 54 proteins -- ribosomes carry out the translation of messenger
RNA into proteins, a core process of the cell. The thousands of proteins per cell, in turn, carry out a vast array of functions, from
digestion to the creation of antibodies. Cells require ribosomes to live.
Jewett likens a ribosome to a chef. The ribosome takes the recipe, encoded in DNA, and makes the meal, or a protein. "We want to
make brand new chefs, or ribosomes," Jewett said. "Then we can alter ribosomes to do new things for us."
"The ability to make ribosomes in vitro in a process that mimics the way biology does it opens new avenues for the study of ribosome
synthesis and assembly, enabling us to better understand and possibly control the translation process," he said. "Our technology also
may enable us in the future to rapidly engineer modified ribosomes with new behaviors and functions, a potentially significant advance
for the synthetic biology field."
The synthesis process developed by Jewett and Church -- termed "integrated synthesis, assembly and translation" (iSAT) technology
-- mimics nature by enabling ribosome synthesis, assembly and function in a single reaction and in the same compartment.
Working with E. coli cells, the researchers combined natural ribosomal proteins with synthetically made ribosomal RNA, which selfassembled in vitro to create semi-synthetic, functional ribosomes.They confirmed the ribosomes were active by assessing their ability
to carry out translation of luciferase, the protein responsible for allowing a firefly to glow. The researchers then showed the ability of
iSAT to make a modified ribosome with a point mutation that mediates resistance to the antibiotic clindamycin.
The researchers next want to synthesize all 57 ribosome parts, including the 54 proteins.
"I'm really excited about where we are," Jewett said. "This study is an important step along the way to synthesizing a complete
ribosome. We will continue to push this work forward."
Jewett and Church, a professor of genetics at Harvard Medical School, are authors of the paper, titled "In Vitro Integration of
Ribosomal RNA Synthesis, Ribosome Assembly, and Translation." Other authors are Brian R. Fritz and Laura E. Timmerman,
graduate students in chemical and biological engineering at Northwestern.
The work was carried out at both Northwestern University and Harvard Medical School.

Reference:
http://www.sciencedaily.com/releases/2013/06/130629164739.htm