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Article ; Online: Keeping mammalian mutation load in check: regulation of the activity of error-prone DNA polymerases by p53 and p21.

Livneh, Zvi

2006 Volume 5, Issue 17, Page(s) 1918–1922

Abstract: To overcome DNA lesions that block replication the cell employs translesion DNA synthesis (TLS) polymerases, a group of low fidelity DNA polymerases that have the capacity to bypass a wide range of DNA lesions. This TLS process is also termed error-prone ...

Abstract	To overcome DNA lesions that block replication the cell employs translesion DNA synthesis (TLS) polymerases, a group of low fidelity DNA polymerases that have the capacity to bypass a wide range of DNA lesions. This TLS process is also termed error-prone repair, due to its inherent mutagenic nature. We have recently shown that the tumor suppressor p53 and the cell cycle inhibitor p21 are global regulators of TLS. When these proteins are missing or nonfunctional, TLS gets out of control: its extent increases to very high levels, and its fidelity decreases, causing an overall increase in mutation load. This may be explained by the loss of selectivity in the bypass of specific DNA lesions by their cognate specialized polymerases, such that lesion bypass continues to a maximum, regardless of the price paid in increased mutations. The p53 and p21 proteins are also required for efficient UV light-induced monoubiquitination of PCNA, which is consistent with a model in which this modification of PCNA is necessary but not sufficient for the normal activity of TLS. This regulation suggests that TLS evolved in mammals as a system that balances gain in survival with a tolerable mutational cost, and that disturbing this balance causes a potentially harmful increase in mutations, which might play a role in carcinogenesis.
MeSH term(s)	Animals ; Cyclin-Dependent Kinase Inhibitor p21/metabolism ; DNA/biosynthesis ; DNA Damage ; DNA Repair ; DNA-Directed DNA Polymerase/metabolism ; Models, Genetic ; Mutation ; Proliferating Cell Nuclear Antigen/metabolism ; Tumor Suppressor Protein p53/metabolism ; Ubiquitin/metabolism
Chemical Substances	Cyclin-Dependent Kinase Inhibitor p21 ; Proliferating Cell Nuclear Antigen ; Tumor Suppressor Protein p53 ; Ubiquitin ; DNA (9007-49-2) ; DNA-Directed DNA Polymerase (EC 2.7.7.7)
Language	English
Publishing date	2006-09-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2146183-1
ISSN	1551-4005 ; 1538-4101 ; 1554-8627
ISSN (online)	1551-4005
ISSN	1538-4101 ; 1554-8627
DOI	10.4161/cc.5.17.3193
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Article: A Highly Sensitive Flow Cytometric Approach to Detect Rare Antigen-Specific T Cells: Development and Comparison to Standard Monitoring Tools.

Dror Levinsky, Meytal / Brenner, Baruch / Yalon, Michal / Levi, Zohar / Livneh, Zvi / Cohen, Zoya / Paz-Elizur, Tamar / Grossman, Rachel / Ram, Zvi / Volovitz, Ilan

Cancers

2023 Volume 15, Issue 3

Abstract: Personalized vaccines against patient-unique tumor-associated antigens represent a promising new approach for cancer immunotherapy. Vaccine efficacy is assessed by quantification of changes in the frequency and/or the activity of antigen-specific T cells. ...

Abstract	Personalized vaccines against patient-unique tumor-associated antigens represent a promising new approach for cancer immunotherapy. Vaccine efficacy is assessed by quantification of changes in the frequency and/or the activity of antigen-specific T cells. Enzyme-linked immunosorbent spot (ELISpot) and flow cytometry (FCM) are methodologies frequently used for assessing vaccine efficacy. We tested these methodologies and found that both ELISpot and standard FCM [monitoring CD3/CD4/CD8/IFNγ/Viability+CD14+CD19 (dump)] demonstrate background IFNγ secretion, which, in many cases, was higher than the antigen-specific signal measured by the respective methodology (frequently ranging around 0.05-0.2%). To detect such weak T-cell responses, we developed an FCM panel that included two early activation markers, 4-1BB (CD137) and CD40L (CD154), in addition to the above-cited markers. These two activation markers have a close to zero background expression and are rapidly upregulated following antigen-specific activation. They enabled the quantification of rare T cells responding to antigens within the assay well. Background IFNγ-positive CD4 T cell frequencies decreased to 0.019% ± 0.028% and CD8 T cells to 0.009% ± 0.013%, which are 19 and 13 times lower, respectively, than without the use of these markers. The presented methodology enables highly sensitive monitoring of T-cell responses to tumor-associated antigens in the very low, but clinically relevant, frequencies.
Language	English
Publishing date	2023-01-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers15030574
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Article ; Online: Effective bubble-based testing for SARS-CoV-2 using swab-pooling.

Cohen, Yuval / Bamberger, Nadav / Mor, Orna / Walfisch, Ronen / Fleishon, Shay / Varkovitzky, Itay / Younger, Asaf / Levi, Danit Oz / Kohn, Yishai / Steinberg, David M / Zeevi, Danny / Erster, Oran / Mendelson, Ella / Livneh, Zvi

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases

2022 Volume 28, Issue 6, Page(s) 859–864

Abstract: Objectives: Despite the success in developing COVID-19 vaccines, containment of the disease is obstructed worldwide by vaccine production bottlenecks, logistics hurdles, vaccine refusal, transmission through unvaccinated children, and the appearance of ... ...

Abstract	Objectives: Despite the success in developing COVID-19 vaccines, containment of the disease is obstructed worldwide by vaccine production bottlenecks, logistics hurdles, vaccine refusal, transmission through unvaccinated children, and the appearance of new viral variants. This underscores the need for effective strategies for identifying carriers/patients, which was the main aim of this study. Methods: We present a bubble-based PCR testing approach using swab-pooling into lysis buffer. A bubble is a cluster of people who can be periodically tested for SARS-CoV-2 by swab-pooling. A positive test of a pool mandates quarantining each of its members, who are then individually tested while in isolation to identify the carrier(s) for further epidemiological contact tracing. Results: We tested an overall sample of 25 831 individuals, divided into 1273 bubbles, with an average size of 20.3 ± 7.7 swabs/test tube, obtaining for all pools (≤37 swabs/pool) a specificity of 97.5% (lower bound 96.6%) and a sensitivity of 86.3% (lower bound 78.2%) and a post hoc analyzed sensitivity of 94.6% (lower bound 86.7%) and a specificity of 97.2% (lower bound 96.2%) in pools with ≤25 swabs, relative to individual testing. Discussion: This approach offers a significant scale-up in sampling and testing throughput and savings in testing cost, without reducing sensitivity or affecting the standard PCR testing laboratory routine. It can be used in school classes, airplanes, hospitals, military units, and workplaces, and may be applicable to future pandemics.
MeSH term(s)	COVID-19/diagnosis ; COVID-19 Testing ; COVID-19 Vaccines ; Child ; Humans ; Pandemics ; RNA, Viral ; SARS-CoV-2/genetics ; Sensitivity and Specificity ; Specimen Handling
Chemical Substances	COVID-19 Vaccines ; RNA, Viral
Language	English
Publishing date	2022-02-17
Publishing country	England
Document type	Journal Article
ZDB-ID	1328418-6
ISSN	1469-0691 ; 1470-9465 ; 1198-743X
ISSN (online)	1469-0691
ISSN	1470-9465 ; 1198-743X
DOI	10.1016/j.cmi.2022.02.016
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Zs.A 4541: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: TENT4A Non-Canonical Poly(A) Polymerase Regulates DNA-Damage Tolerance via Multiple Pathways That Are Mutated in Endometrial Cancer.

Swain, Umakanta / Friedlander, Gilgi / Sehrawat, Urmila / Sarusi-Portuguez, Avital / Rotkopf, Ron / Ebert, Charlotte / Paz-Elizur, Tamar / Dikstein, Rivka / Carell, Thomas / Geacintov, Nicholas E / Livneh, Zvi

International journal of molecular sciences

2021 Volume 22, Issue 13

Abstract: TENT4A (PAPD7) is a non-canonical poly(A) polymerase, of which little is known. Here, we show that TENT4A regulates multiple biological pathways and focuses on its multilayer regulation of translesion DNA synthesis (TLS), in which error-prone DNA ... ...

Abstract	TENT4A (PAPD7) is a non-canonical poly(A) polymerase, of which little is known. Here, we show that TENT4A regulates multiple biological pathways and focuses on its multilayer regulation of translesion DNA synthesis (TLS), in which error-prone DNA polymerases bypass unrepaired DNA lesions. We show that TENT4A regulates mRNA stability and/or translation of DNA polymerase η and RAD18 E3 ligase, which guides the polymerase to replication stalling sites and monoubiquitinates PCNA, thereby enabling recruitment of error-prone DNA polymerases to damaged DNA sites. Remarkably, in addition to the effect on RAD18 mRNA stability via controlling its poly(A) tail, TENT4A indirectly regulates RAD18 via the tumor suppressor CYLD and via the long non-coding antisense RNA
MeSH term(s)	Blotting, Western ; Cell Line, Tumor ; Chromosomal Proteins, Non-Histone/genetics ; Chromosomal Proteins, Non-Histone/metabolism ; Computational Biology ; DNA Damage/genetics ; DNA Damage/physiology ; DNA Repair/genetics ; DNA Repair/physiology ; DNA Replication/genetics ; DNA Replication/physiology ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; DNA-Directed DNA Polymerase/genetics ; DNA-Directed DNA Polymerase/metabolism ; Endometrial Neoplasms/genetics ; Endometrial Neoplasms/metabolism ; Female ; HEK293 Cells ; Humans ; Immunoprecipitation ; MCF-7 Cells ; Mutation/genetics ; Polymerase Chain Reaction ; Polynucleotide Adenylyltransferase/genetics ; Polynucleotide Adenylyltransferase/metabolism ; RNA Stability/genetics ; RNA Stability/physiology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Ubiquitin-Protein Ligases/genetics ; Ubiquitin-Protein Ligases/metabolism ; Ubiquitination/genetics ; Ubiquitination/physiology
Chemical Substances	Chromosomal Proteins, Non-Histone ; DNA-Binding Proteins ; PAXIP1 protein, human ; RAD18 protein, human ; RNA, Messenger ; Ubiquitin-Protein Ligases (EC 2.3.2.27) ; Polynucleotide Adenylyltransferase (EC 2.7.7.19) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; TENT4A protein, human (EC 2.7.7.7)
Language	English
Publishing date	2021-06-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22136957
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Article ; Online: Recurrent deletions in clonal hematopoiesis are driven by microhomology-mediated end joining.

Feldman, Tzah / Bercovich, Akhiad / Moskovitz, Yoni / Chapal-Ilani, Noa / Mitchell, Amanda / Medeiros, Jessie J F / Biezuner, Tamir / Kaushansky, Nathali / Minden, Mark D / Gupta, Vikas / Milyavsky, Michael / Livneh, Zvi / Tanay, Amos / Shlush, Liran I

Nature communications

2021 Volume 12, Issue 1, Page(s) 2455

Abstract: The mutational mechanisms underlying recurrent deletions in clonal hematopoiesis are not entirely clear. In the current study we inspect the genomic regions around recurrent deletions in myeloid malignancies, and identify microhomology-based signatures ... ...

Abstract	The mutational mechanisms underlying recurrent deletions in clonal hematopoiesis are not entirely clear. In the current study we inspect the genomic regions around recurrent deletions in myeloid malignancies, and identify microhomology-based signatures in CALR, ASXL1 and SRSF2 loci. We demonstrate that these deletions are the result of double stand break repair by a PARP1 dependent microhomology-mediated end joining (MMEJ) pathway. Importantly, we provide evidence that these recurrent deletions originate in pre-leukemic stem cells. While DNA polymerase theta (POLQ) is considered a key component in MMEJ repair, we provide evidence that pre-leukemic MMEJ (preL-MMEJ) deletions can be generated in POLQ knockout cells. In contrast, aphidicolin (an inhibitor of replicative polymerases and replication) treatment resulted in a significant reduction in preL-MMEJ. Altogether, our data indicate an association between POLQ independent MMEJ and clonal hematopoiesis and elucidate mutational mechanisms involved in the very first steps of leukemia evolution.
MeSH term(s)	Aphidicolin/pharmacology ; Calreticulin/genetics ; Clonal Hematopoiesis/genetics ; DNA Breaks, Double-Stranded ; DNA End-Joining Repair/genetics ; DNA-Directed DNA Polymerase/drug effects ; DNA-Directed DNA Polymerase/genetics ; Enzyme Inhibitors/pharmacology ; Humans ; Leukemia, Myeloid/genetics ; Myeloid Progenitor Cells ; Poly (ADP-Ribose) Polymerase-1/metabolism ; Repressor Proteins/genetics ; Sequence Deletion/genetics ; Serine-Arginine Splicing Factors/genetics ; DNA Polymerase theta
Chemical Substances	ASXL1 protein, human ; CALR protein, human ; Calreticulin ; Enzyme Inhibitors ; Repressor Proteins ; SRSF2 protein, human (147153-65-9) ; Serine-Arginine Splicing Factors (170974-22-8) ; Aphidicolin (38966-21-1) ; PARP1 protein, human (EC 2.4.2.30) ; Poly (ADP-Ribose) Polymerase-1 (EC 2.4.2.30) ; DNA-Directed DNA Polymerase (EC 2.7.7.7)
Language	English
Publishing date	2021-04-28
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-021-22803-y
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Article: Translesion DNA synthesis across non-DNA segments in cultured human cells.

Adar, Sheera / Livneh, Zvi

DNA repair

2006 Volume 5, Issue 4, Page(s) 479–490

Abstract: DNA lesions that have escaped DNA repair are tolerated via translesion DNA synthesis (TLS), carried out by specialized error-prone DNA polymerases. To evaluate the robustness of the TLS system in human cells, we examined its ability to cope with foreign ... ...

Abstract	DNA lesions that have escaped DNA repair are tolerated via translesion DNA synthesis (TLS), carried out by specialized error-prone DNA polymerases. To evaluate the robustness of the TLS system in human cells, we examined its ability to cope with foreign non-DNA stretches of 3 or 12 methylene residues, using a gap-lesion plasmid assay system. We found that both the trimethylene and dodecamethylene inserts were bypassed with significant efficiencies in human cells, using both misinsertion and misalignment mechanisms. TLS across these non-DNA segments was aphidicolin-sensitive, and did not require poleta. In vitro primer extension assays showed that purified poleta, polkappa and poliota were each capable of inserting each of the four nucleotides opposite the trimethylene chain, but only poleta and polkappa could fully bypass it. Poleta and poliota, but not polkappa, could also insert each of the four nucleotides opposite the dodecamethylene chain, but all three polymerases were severely blocked by this lesion. The ability of TLS polymerases to insert nucleotides opposite a hydrocarbon chain, despite the lack of any similarity to DNA, suggests that they may act via a mode of transient and local template-independent polymerase activity, and highlights the robustness of the TLS system in human cells.
MeSH term(s)	Animals ; Cells, Cultured ; Cyclopropanes/chemistry ; DNA/biosynthesis ; DNA/chemistry ; DNA/metabolism ; DNA Replication ; DNA-Directed DNA Polymerase/isolation & purification ; DNA-Directed DNA Polymerase/metabolism ; Humans ; Hydrocarbons, Acyclic/metabolism ; Models, Molecular ; Nucleic Acid Conformation ; Nucleotides/metabolism ; Substrate Specificity
Chemical Substances	Cyclopropanes ; Hydrocarbons, Acyclic ; Nucleotides ; DNA (9007-49-2) ; cyclopropane (99TB643425) ; DNA polymerase iota (EC 2.7.7.-) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; POLK protein, human (EC 2.7.7.7) ; Rad30 protein (EC 2.7.7.7)
Language	English
Publishing date	2006-04-08
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2071608-4
ISSN	1568-7856 ; 1568-7864
ISSN (online)	1568-7856
ISSN	1568-7864
DOI	10.1016/j.dnarep.2006.01.001
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Article ; Online: Author Correction: Mutational signatures reveal the role of RAD52 in p53-independent p21-driven genomic instability.

Genome biology

2022 Volume 23, Issue 1, Page(s) 107

Language	English
Publishing date	2022-04-28
Publishing country	England
Document type	Published Erratum
ZDB-ID	2040529-7
ISSN	1474-760X ; 1474-760X
ISSN (online)	1474-760X
ISSN	1474-760X
DOI	10.1186/s13059-022-02678-y
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Article ; Online: DNA sequence context greatly affects the accuracy of bypass across an ultraviolet light 6-4 photoproduct in mammalian cells.

Shriber, Pola / Leitner-Dagan, Yael / Geacintov, Nicholas / Paz-Elizur, Tamar / Livneh, Zvi

Mutation research

2015 Volume 780, Page(s) 71–76

Abstract: Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism carried out by low-fidelity DNA polymerases that bypass DNA lesions, which overcomes replication stalling. Despite the miscoding nature of most common DNA lesions, several of them are ... ...

Abstract	Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism carried out by low-fidelity DNA polymerases that bypass DNA lesions, which overcomes replication stalling. Despite the miscoding nature of most common DNA lesions, several of them are bypassed in mammalian cells in a relatively accurate manner, which plays a key role maintaining a low mutation load. Whereas it is generally agreed that TLS across the major UV and sunlight induced DNA lesion, the cyclobutane pyrimidine dimer (CPD), is accurate, there were conflicting reports on whether the same is true for the thymine-thymine pyrimidine-pyrimidone(6-4) ultraviolet light photoproduct (TT6-4PP), which represents the second most common class of UV lesions. Using a TLS assay system based on gapped plasmids carrying site-specific TT6-4PP lesions in defined sequence contexts we show that the DNA sequence context markedly affected both the extent and accuracy of TLS. The sequence exhibiting higher TLS exhibited also higher error-frequency, caused primarily by semi-targeted mutations, at the nearest nucleotides flanking the lesion. Our results resolve the discrepancy reported on TLS across TT6-4PP, and suggest that TLS is more accurate in human cells than in mouse cells.
MeSH term(s)	Animals ; Cell Line, Transformed ; DNA Damage ; Humans ; Mice ; Mutation ; Pyrimidine Dimers/genetics ; Pyrimidine Dimers/metabolism ; Species Specificity ; Ultraviolet Rays/adverse effects
Chemical Substances	Pyrimidine Dimers
Language	English
Publishing date	2015-10
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	206607-5
ISSN	1873-135X ; 1383-5718 ; 0027-5107 ; 0165-1110 ; 0165-1161 ; 0165-7992 ; 0921-8777 ; 0165-1218 ; 1383-5726 ; 0167-8817 ; 0921-8734 ; 1383-5742
ISSN (online)	1873-135X
ISSN	1383-5718 ; 0027-5107 ; 0165-1110 ; 0165-1161 ; 0165-7992 ; 0921-8777 ; 0165-1218 ; 1383-5726 ; 0167-8817 ; 0921-8734 ; 1383-5742
DOI	10.1016/j.mrfmmm.2015.08.002
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Article ; Online: Multiple two-polymerase mechanisms in mammalian translesion DNA synthesis.

Livneh, Zvi / Ziv, Omer / Shachar, Sigal

Cell cycle (Georgetown, Tex.)

2010 Volume 9, Issue 4, Page(s) 729–735

Abstract: The encounter of replication forks with DNA lesions may lead to fork arrest and/or the formation of single-stranded gaps. A major strategy to cope with these replication irregularities is translesion DNA synthesis (TLS), in which specialized error-prone ... ...

Abstract	The encounter of replication forks with DNA lesions may lead to fork arrest and/or the formation of single-stranded gaps. A major strategy to cope with these replication irregularities is translesion DNA synthesis (TLS), in which specialized error-prone DNA polymerases bypass the blocking lesions. Recent studies suggest that TLS across a particular DNA lesion may involve as many as four different TLS polymerases, acting in two-polymerase reactions in which insertion by a particular polymerase is followed by extension by another polymerase. Insertion determines the accuracy and mutagenic specificity of the TLS reaction, and is carried out by one of several polymerases such as poleta, polkappa or poliota. In contrast, extension is carried out primarily by polzeta. In cells from XPV patients, which are deficient in TLS across cyclobutane pyrimidine dimers (CPD) due to a deficiency in poleta, TLS is carried out by at least two backup reactions each involving two polymerases: One reaction involves polkappa and polzeta, and the other poliota and polzeta. These mechanisms may also assist poleta in normal cells under an excessive amount of UV lesions.
MeSH term(s)	DNA/biosynthesis ; DNA Repair ; DNA-Directed DNA Polymerase/metabolism ; Humans
Chemical Substances	DNA (9007-49-2) ; DNA polymerase iota (EC 2.7.7.-) ; DNA polymerase zeta (EC 2.7.7.-) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; POLK protein, human (EC 2.7.7.7) ; Rad30 protein (EC 2.7.7.7)
Language	English
Publishing date	2010-02-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2146183-1
ISSN	1551-4005 ; 1538-4101 ; 1554-8627
ISSN (online)	1551-4005
ISSN	1538-4101 ; 1554-8627
DOI	10.4161/cc.9.4.10727
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Article ; Online: Quantitative measurement of translesion DNA synthesis in mammalian cells.

Ziv, Omer / Diamant, Noam / Shachar, Sigal / Hendel, Ayal / Livneh, Zvi

Methods in molecular biology (Clifton, N.J.)

2012 Volume 920, Page(s) 529–542

Abstract: Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism, in which specialized low-fidelity DNA polymerases bypass lesions that interfere with replication. This process is inherently mutagenic due to the miscoding nature of DNA lesions, but it ...

Abstract	Translesion DNA synthesis (TLS) is a DNA damage tolerance mechanism, in which specialized low-fidelity DNA polymerases bypass lesions that interfere with replication. This process is inherently mutagenic due to the miscoding nature of DNA lesions, but it prevents double strand breaks, genome instability, and cancer. We describe here a quantitative method for measuring TLS in mammalian cells, based on non-replicating plasmids that carry a defined and site-specific DNA lesion in a single-stranded DNA region opposite a gap. The assay is responsive to the cellular composition of TLS DNA polymerases, and TLS regulators. It can be used with a broad variety of cultured mammalian cells, and is amenable to RNAi gene silencing, making it a useful tool in the study of TLS in mammalian cells.
MeSH term(s)	Animals ; Base Sequence ; Cells, Cultured ; Chromosomes/genetics ; DNA Damage ; DNA, Single-Stranded/biosynthesis ; DNA, Single-Stranded/genetics ; Genetic Techniques ; Genetic Vectors/genetics ; Oligodeoxyribonucleotides/biosynthesis ; Oligodeoxyribonucleotides/genetics ; Plasmids/genetics
Chemical Substances	DNA, Single-Stranded ; Oligodeoxyribonucleotides
Language	English
Publishing date	2012
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-61779-998-3_35
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