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Abaffy, Tatjana

Positions:

Assistant Research Professor in Molecular Genetics and Microbiology

Molecular Genetics and Microbiology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 2000

Ph.D. — University of Auckland (New Zealand)

Grants:

Peripheral Odor Coding in Mammals

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Assistant Research Professor
Start Date
April 01, 2015
End Date
March 31, 2020

Olfactory moding in mammals

Administered By
Molecular Genetics and Microbiology
AwardedBy
University of Pennsylvania
Role
Assistant Research Professor
Start Date
August 01, 2015
End Date
July 31, 2018

Publications:

Discovery of novel ligands for mouse olfactory receptor MOR42-3 using an in silico screening approach and in vitro validation.

The ligands for many olfactory receptors remain largely unknown despite successful heterologous expression of these receptors. Understanding the molecular receptive range of olfactory receptors and deciphering the olfactory recognition code are hampered by the huge number of odorants and large number of olfactory receptors, as well as the complexity of their combinatorial coding. Here, we present an in silico screening approach to find additional ligands for a mouse olfactory receptor that allows improved definition of its molecular receptive range. A virtual library of 574 odorants was screened against a mouse olfactory receptor MOR42-3. We selected the top 20 candidate ligands using two different scoring functions. These 40 odorant candidate ligands were then tested in vitro using the Xenopus oocyte heterologous expression system and two-electrode voltage clamp electrophysiology. We experimentally confirmed 22 of these ligands. The candidate ligands were screened for both agonist and antagonist activity. In summary, we validated 19 agonists and 3 antagonists. Two of the newly identified antagonists were of low potency. Several previously known ligands (mono- and dicarboxylic acids) are also confirmed in this study. However, some of the newly identified ligands were structurally dissimilar compounds with various functional groups belonging to aldehydes, phenyls, alkenes, esters and ethers. The high positive predictive value of our in silico approach is promising. We believe that this approach can be used for initial deorphanization of olfactory receptors as well as for future comprehensive studies of molecular receptive range of olfactory receptors.

Authors
Bavan, S; Sherman, B; Luetje, CW; Abaffy, T
MLA Citation
Bavan, S, Sherman, B, Luetje, CW, and Abaffy, T. "Discovery of novel ligands for mouse olfactory receptor MOR42-3 using an in silico screening approach and in vitro validation." PloS one 9.3 (January 2014): e92064-.
PMID
24637889
Source
epmc
Published In
PloS one
Volume
9
Issue
3
Publish Date
2014
Start Page
e92064
DOI
10.1371/journal.pone.0092064

A non-invasive method for in vivo skin volatile compounds sampling.

The use of volatile organic compounds (VOCs) emanating from human skin presents great potential for skin disease diagnosis. These compounds are emitted at very low concentrations. Thus, the sampling preparation step needs to be implemented before gas chromatography-mass spectrometry (GC-MS) analysis. In this work, a simple, non-invasive headspace sampling method for volatile compounds emanating from human skin is presented, using thin film as the extraction phase format. The proposed method was evaluated in terms of reproducibility, membrane size, extraction mode and storage conditions. First, the in vial sampling showed an intra- and inter-membrane RSD% less than 9.8% and 8.2%, respectively, which demonstrated that this home-made skin volatiles sampling device was highly reproducible with regard to intra-, inter-membrane sampling. The in vivo sampling was influenced not only by the skin metabolic status, but also by environmental conditions. The developed sampling set-up (or "membrane sandwich") was used to compare two different modes of sampling: headspace and direct sampling. Results demonstrated that headspace sampling had significantly reduced background signal intensity, indicating minimized contamination from the skin surface. In addition, membrane storage conditions both before and after sampling were fully investigated. Membranes stored in dry ice for up to 72 h after collection were tested and showed no or minimal change in volatile profiles. This novel skin volatile compounds sampling approach coupled with gas chromatography-mass spectrometry (GC-MS) can achieve reproducible analysis. This technique was applied to identify the biomarkers of garlic intake and alcohol ingestion. Dimethyl sulphone, allyl methyl sulfide and allyl mercaptan, as metabolites of garlic intake, were detected. In addition, alcohol released from skin was also detected using our "membrane-sandwich" sampling. Using the same approach, we analyzed skin VOCs from upper back, forearm and back thigh regions of the body. Our results show that different body locations share a number of common compounds (27/99). The area with most compounds detected was the upper back skin region, where the density of sebaceous glands is the highest.

Authors
Jiang, R; Cudjoe, E; Bojko, B; Abaffy, T; Pawliszyn, J
MLA Citation
Jiang, R, Cudjoe, E, Bojko, B, Abaffy, T, and Pawliszyn, J. "A non-invasive method for in vivo skin volatile compounds sampling." Analytica chimica acta 804 (December 2013): 111-119.
PMID
24267071
Source
epmc
Published In
Analytica Chimica Acta
Volume
804
Publish Date
2013
Start Page
111
End Page
119
DOI
10.1016/j.aca.2013.09.056

Comparative analysis of volatile metabolomics signals from melanoma and benign skin: a pilot study.

The analysis of volatile organic compounds (VOC) as biomarkers of cancer is both promising and challenging. In this pilot study, we used an untargeted approach to compare volatile metabolomic signatures of melanoma and matched control non-neoplastic skin from the same patient. VOC from fresh (non-fixed) biopsied tissue were collected using the headspace solid phase micro extraction method (HS SPME) and analyzed by gas chromatography and mass spectrometry (GCMS). We applied the XCMS analysis platform and MetaboAnalyst software to reveal many differentially expressed metabolic features. Our analysis revealed increased levels of lauric acid (C12:0) and palmitic acid (C16:0) in melanoma. The identity of these compounds was confirmed by comparison with chemical standards. Increased levels of these fatty acids are likely to be a consequence of up-regulated de novo lipid synthesis, a known characteristic of cancer. Increased oxidative stress is likely to cause an additional increase in lauric acid. Implementation of this study design on larger number of cases will be necessary for the future metabolomics biomarker discovery applications.

Authors
Abaffy, T; Möller, MG; Riemer, DD; Milikowski, C; DeFazio, RA
MLA Citation
Abaffy, T, Möller, MG, Riemer, DD, Milikowski, C, and DeFazio, RA. "Comparative analysis of volatile metabolomics signals from melanoma and benign skin: a pilot study." Metabolomics : Official journal of the Metabolomic Society 9.5 (January 2013): 998-1008.
PMID
24039618
Source
epmc
Published In
Metabolomics
Volume
9
Issue
5
Publish Date
2013
Start Page
998
End Page
1008
DOI
10.1007/s11306-013-0523-z

A protocol for characterizing the impact of collateral flow after distal middle cerebral artery occlusion.

In humans and in animal models of stroke, collateral blood flow between territories of the major pial arteries has a profound impact on cortical infarct size. However, there is a gap in our understanding of the genetic determinants of collateral formation and flow, as well as the signaling pathways and neurovascular interactions regulating this flow. Previous studies have demonstrated that collateral flow between branches of the anterior cerebral artery (ACA) and the middle cerebral artery (MCA) can protect mouse cortex from infarction after middle cerebral artery occlusion. Because the number and diameter of collaterals varies among mouse strains and after transgenic manipulations, a combination of methods is required to control for these variations. Here, we report an inexpensive approach to characterizing the cerebrovascular anatomy, and in vivo monitoring of cerebral blood flow as well. Further, we introduce a new, minimally invasive method for the occlusion of distal MCA branches. These methods will permit a new generation of studies on the mechanisms regulating collateral remodeling and cortical blood flow after stroke.

Authors
Defazio, RA; Levy, S; Morales, CL; Levy, RV; Dave, KR; Lin, HW; Abaffy, T; Watson, BD; Perez-Pinzon, MA; Ohanna, V
MLA Citation
Defazio, RA, Levy, S, Morales, CL, Levy, RV, Dave, KR, Lin, HW, Abaffy, T, Watson, BD, Perez-Pinzon, MA, and Ohanna, V. "A protocol for characterizing the impact of collateral flow after distal middle cerebral artery occlusion." Translational stroke research 2.1 (March 2011): 112-127.
PMID
21593993
Source
epmc
Published In
Translational Stroke Research
Volume
2
Issue
1
Publish Date
2011
Start Page
112
End Page
127
DOI
10.1007/s12975-010-0044-2

A case report - Volatile metabolomic signature of malignant melanoma using matching skin as a control.

Melanoma is the most serious form of skin cancer. The quest for melanoma diagnostic biomarkers is paramount since early detection of melanoma and surgical excision represent the only effective treatment of this capricious disease. Our recent study tested the hypothesis that melanoma forms a unique volatile signature that is different than control, healthy tissue. Here, we are reporting a case study, the analysis of the volatile metabolic signature of a malignant melanoma using matched, non-neoplastic skin tissue from the same patient as a control. This is a significant improvement in the methodology, since it is well known that diet, skin type, genetic background, age, sex and environment all contribute to individual variation in the skin volatile signature. In the present study, we have identified 32 volatile compounds; 9 volatile compounds were increased in melanoma when compared to normal skin and 23 volatile compounds were detected only in melanoma and not in normal skin. Out of these 32 compounds, 10 have been reported previously by our group, thus confirming our results and adding additional confidence in our untargeted metabolomics approach for detection of melanoma biomarkers.

Authors
Abaffy, T; Möller, M; Riemer, DD; Milikowski, C; Defazio, RA
MLA Citation
Abaffy, T, Möller, M, Riemer, DD, Milikowski, C, and Defazio, RA. "A case report - Volatile metabolomic signature of malignant melanoma using matching skin as a control." Journal of cancer science & therapy 3.6 (January 2011): 140-144.
PMID
22229073
Source
epmc
Published In
Journal of Cancer Science and Therapy
Volume
3
Issue
6
Publish Date
2011
Start Page
140
End Page
144
DOI
10.4172/1948-5956.1000076

The location of olfactory receptors within olfactory epithelium is independent of odorant volatility and solubility.

BACKGROUND: Our objective was to study the pattern of olfactory receptor expression within the dorsal and ventral regions of the mouse olfactory epithelium. We hypothesized that olfactory receptors were distributed based on the chemical properties of their ligands: e.g. receptors for polar, hydrophilic and weakly volatile odorants would be present in the dorsal region of olfactory epithelium; while receptors for non-polar, more volatile odorants would be distributed to the ventral region. To test our hypothesis, we used micro-transplantation of cilia-enriched plasma membranes derived from dorsal or ventral regions of the olfactory epithelium into Xenopus oocytes for electrophysiological characterization against a panel of 100 odorants. FINDINGS: Odorants detected by ORs from the dorsal and ventral regions showed overlap in volatility and water solubility. We did not find evidence for a correlation between the solubility and volatility of odorants and the functional expression of olfactory receptors in the dorsal or ventral region of the olfactory epithelia. CONCLUSIONS: No simple clustering or relationship between chemical properties of odorants could be associated with the different regions of the olfactory epithelium. These results suggest that the location of ORs within the epithelium is not organized based on the physico-chemical properties of their ligands.

Authors
Abaffy, T; Defazio, AR
MLA Citation
Abaffy, T, and Defazio, AR. "The location of olfactory receptors within olfactory epithelium is independent of odorant volatility and solubility." BMC research notes 4 (January 2011): 137-.
PMID
21548958
Source
epmc
Published In
BMC Research Notes
Volume
4
Publish Date
2011
Start Page
137
DOI
10.1186/1756-0500-4-137

Differential volatile signatures from skin, naevi and melanoma: a novel approach to detect a pathological process.

Early detection of melanoma is of great importance to reduce mortality. Discovering new melanoma biomarkers would improve early detection and diagnosis. Here, we present a novel approach to detect volatile compounds from skin.We used Head Space Solid Phase Micro-Extraction (HS-SPME) and gas chromatography/mass spectrometry (GC/MS) to identify volatile signatures from melanoma, naevi and skin samples. We hypothesized that the metabolic state of tissue alters the profile of volatile compounds. Volatiles released from fresh biopsy tissue of melanoma and benign naevus were compared based on their difference in frequency distribution and their expression level. We also analyzed volatile profiles from frozen tissue, including skin and melanoma.Three volatiles, 4-methyl decane, dodecane and undecane were preferentially expressed in both fresh and frozen melanoma, indicating that they are candidate biomarkers. Twelve candidate biomarkers evaluated by fuzzy logic analysis of frozen samples distinguished melanoma from skin with 89% sensitivity and 90% specificity. Our results demonstrate proof-of-principle that there is differential expression of volatiles in melanoma. Our volatile metabolomic approach will lead to a better understanding of melanoma and can enable development of new diagnostic and treatment strategies based on altered metabolism.

Authors
Abaffy, T; Duncan, R; Riemer, DD; Tietje, O; Elgart, G; Milikowski, C; DeFazio, RA
MLA Citation
Abaffy, T, Duncan, R, Riemer, DD, Tietje, O, Elgart, G, Milikowski, C, and DeFazio, RA. "Differential volatile signatures from skin, naevi and melanoma: a novel approach to detect a pathological process." PloS one 5.11 (November 4, 2010): e13813-.
PMID
21079799
Source
epmc
Published In
PloS one
Volume
5
Issue
11
Publish Date
2010
Start Page
e13813
DOI
10.1371/journal.pone.0013813

Genetic elucidation of human hyperosmia to isovaleric acid.

The genetic basis of odorant-specific variations in human olfactory thresholds, and in particular of enhanced odorant sensitivity (hyperosmia), remains largely unknown. Olfactory receptor (OR) segregating pseudogenes, displaying both functional and nonfunctional alleles in humans, are excellent candidates to underlie these differences in olfactory sensitivity. To explore this hypothesis, we examined the association between olfactory detection threshold phenotypes of four odorants and segregating pseudogene genotypes of 43 ORs genome-wide. A strong association signal was observed between the single nucleotide polymorphism variants in OR11H7P and sensitivity to the odorant isovaleric acid. This association was largely due to the low frequency of homozygous pseudogenized genotype in individuals with specific hyperosmia to this odorant, implying a possible functional role of OR11H7P in isovaleric acid detection. This predicted receptor-ligand functional relationship was further verified using the Xenopus oocyte expression system, whereby the intact allele of OR11H7P exhibited a response to isovaleric acid. Notably, we also uncovered another mechanism affecting general olfactory acuity that manifested as a significant inter-odorant threshold concordance, resulting in an overrepresentation of individuals who were hyperosmic to several odorants. An involvement of polymorphisms in other downstream transduction genes is one possible explanation for this observation. Thus, human hyperosmia to isovaleric acid is a complex trait, contributed to by both receptor and other mechanisms in the olfactory signaling pathway.

Authors
Menashe, I; Abaffy, T; Hasin, Y; Goshen, S; Yahalom, V; Luetje, CW; Lancet, D
MLA Citation
Menashe, I, Abaffy, T, Hasin, Y, Goshen, S, Yahalom, V, Luetje, CW, and Lancet, D. "Genetic elucidation of human hyperosmia to isovaleric acid." PLoS biology 5.11 (October 2007): e284-.
PMID
17973576
Source
epmc
Published In
PLoS biology
Volume
5
Issue
11
Publish Date
2007
Start Page
e284
DOI
10.1371/journal.pbio.0050284

The molecular basis for ligand specificity in a mouse olfactory receptor: a network of functionally important residues.

Sequence differences between members of the mouse olfac-tory receptor MOR42 subfamily (MOR42-3 and MOR42-1) are likely to be the basis for variation in ligand binding preference among these receptors. We investigated the specificity of MOR42-3 for a variety of dicarboxylic acids. We used site-directed mutagenesis, guided by homology modeling and ligand docking studies, to locate functionally important residues. Receptors were expressed in Xenopus oocytes and assayed using high throughput electrophysiology. The importance of the Val-113 residue, located deep within the receptor, was analyzed in the context of interhelical interactions. We also screened additional residues predicted to be involved in ligand binding site, based on comparison of ortholog/paralog pairs from the mouse and human olfactory receptor genomes (Man, O., Gilad, Y., and Lancet, D. (2004) Protein Sci. 13, 240-254). A network of 8 residues in transmembrane domains III, V, and VI was identified. These residues form part of the ligand binding pocket of MOR42-3. C12 dicarboxylic acid did not activate the receptor in our functional assay, yet our docking simulations predicted its binding site in MOR42-3. Binding without activation implied that C12 dicarboxylic acid might act as an antagonist. In our functional assay, C12 dicarboxylic acid did indeed act as an antagonist of MOR42-3, in agreement with molecular docking studies. Our results demonstrate a powerful approach based on the synergy between computational predictions and physiological assays.

Authors
Abaffy, T; Malhotra, A; Luetje, CW
MLA Citation
Abaffy, T, Malhotra, A, and Luetje, CW. "The molecular basis for ligand specificity in a mouse olfactory receptor: a network of functionally important residues." The Journal of biological chemistry 282.2 (January 2007): 1216-1224.
PMID
17114180
Source
epmc
Published In
The Journal of biological chemistry
Volume
282
Issue
2
Publish Date
2007
Start Page
1216
End Page
1224
DOI
10.1074/jbc.m609355200

Functional analysis of a mammalian odorant receptor subfamily.

Phylogenetic analysis groups mammalian odorant receptors into two broad classes and numerous subfamilies. These subfamilies are proposed to reflect functional organization. Testing this idea requires an assay allowing detailed functional characterization of odorant receptors. Here we show that a variety of Class I and Class II mouse odorant receptors can be functionally expressed in Xenopus laevis oocytes. Receptor constructs included the N-terminal 20 residues of human rhodopsin and were co-expressed with Galphaolf and the cystic fibrosis transmembrane regulator to allow electrophysiological measurement of receptor responses. For most mouse odorant receptors tested, these conditions were sufficient for functional expression. Co-expression of accessory proteins was required to allow functional surface expression of some mouse odorant receptors. We used this assay to examine the receptive ranges of all members of the mouse odorant receptor 42 (MOR42) subfamily. MOR42-1 responded to dicarboxylic acids, preferring a 10-12 carbon chain length. MOR42-2 responded to monocarboxylic acids (7-10 carbons). MOR42-3 responded to dicarboxylic acids (8-10 carbons) and monocarboxylic acids (10-12 carbons). Thus, the receptive range of each receptor was unique. However, overlap between the individual receptive ranges suggests that the members of this subfamily form one contiguous subfamily receptive range, suggesting that odorant receptor subfamilies do constitute functional units.

Authors
Abaffy, T; Matsunami, H; Luetje, CW
MLA Citation
Abaffy, T, Matsunami, H, and Luetje, CW. "Functional analysis of a mammalian odorant receptor subfamily." J Neurochem 97.5 (June 2006): 1506-1518.
PMID
16606354
Source
pubmed
Published In
Journal of Neurochemistry
Volume
97
Issue
5
Publish Date
2006
Start Page
1506
End Page
1518
DOI
10.1111/j.1471-4159.2006.03859.x

GSK3 involvement in amylin signaling in isolated rat soleus muscle.

Amylin can evoke insulin resistance by antagonizing insulin in a non-competitive manner. Here, we investigated the glycogenolytic effect of amylin in isolated skeletal muscle and compared it to the effects of a calcitonin gene-related peptide (CGRP). Amylin alone had no statistically significant effect on glucose transport. However, amylin decreased insulin-stimulated glucose transport by about 30%. The involvement of cAMP could not be detected at the concentrations shown to promote glycogenolysis. Previously, it has been shown that increased glycogen synthase kinase 3 (GSK3) activity plays a role in insulin resistance. Here, the ratio of GSK3 alpha:beta isoforms in rat soleus was found to be 1.2:1. We found that amylin increased GSK3alpha activity, which in turn led to increased phosphorylation of glycogen synthase and decreased glycogen synthesis de novo.

Authors
Abaffy, T; Cooper, GJS
MLA Citation
Abaffy, T, and Cooper, GJS. "GSK3 involvement in amylin signaling in isolated rat soleus muscle." Peptides 25.12 (December 2004): 2119-2125.
PMID
15572200
Source
epmc
Published In
Peptides
Volume
25
Issue
12
Publish Date
2004
Start Page
2119
End Page
2125
DOI
10.1016/j.peptides.2004.08.016

Adenylyl cyclase expression and modulation of cAMP in rat taste cells

cAMP is a second messenger implicated in sensory transduction for taste. The identity of adenylyl cyclase (AC) in taste cells has not been explored. We have employed RT-PCR to identify the AC isoforms present in taste cells and found that AC 4, 6, and 8 are expressed as mRNAs in taste tissue. These proteins are also expressed in a subset of taste cells as revealed by immunohistochemistry. Alterations of cAMP concentrations are associated with transduction of taste stimuli of several classes. The involvement of particular ACs in this modulation has not been investigated. We demonstrate that glutamate, which is a potent stimulus eliciting a taste quality termed umami, causes a decrease in cAMP in forskolin-treated taste cells. The potentiation of this response by inosine monophosphate, the lack of response to D-glutamate, and the lack of response to umami stimuli in nonsensory lingual epithelium all suggest that the cAMP modulation represents umami taste transduction. Because cAMP downregulation via ACs can be mediated through Gα i proteins, we examined the colocalization of the detected ACs with Gα i proteins and found that 66% of AC8 immunopositive taste cells are also positive for gustducin, a taste-specific Gα i protein. Whether AC8 is directly involved in signal transduction of umami taste remains to be established.

Authors
Abaffy, T; Trubey, KR; Chaudhari, N
MLA Citation
Abaffy, T, Trubey, KR, and Chaudhari, N. "Adenylyl cyclase expression and modulation of cAMP in rat taste cells." American Journal of Physiology - Cell Physiology 284.6 53-6 (June 1, 2003).
Source
scopus
Published In
American journal of physiology. Cell physiology
Volume
284
Issue
6 53-6
Publish Date
2003

Troponin T and serum enzymes in monitoring open heart surgery

The values of Troponin T (TnT), a structurally bound protein found in striated muscle cells, and of a group of enzymes were investigated as diagnostic criteria for acute myocardial infarction (AMI) following open heart surgery. In 16 patients subjected to open heart surgery, peripheral venous blood was collected over 5-70 h after surgery. TnT was measured by an enzyme immunoassay (Elisa, Boehringer Mannheim), CK-MB by the immunoinhibition method (Boehringer Mannheim), and total CK, LDH and AST were measured by standard spectrophotometric methods (Herbos, Sisak). The values of CK, LDH and AST were significantly above the reference range (p < 0.05) throughout the measurement, while the CK-MB values were rather inconsistent, sometimes significantly above and sometimes close to the upper reference limit. The TnT values were always slightly above the upper reference limit, but this increase was not statistically significant (p < 0.05). Since the outcome of open heart surgery in the patients was good, the data suggest that the enzyme pattern, being a consequence of abundant enzyme release from the heart tissue and blood cells during the surgery itself, cannot be taken as a reliable diagnostic parameter for AIM. TnT, with its slight increase, could be used as a sensitive and reliable parameter in monitoring the outcome of open heart surgery.

Authors
Topic, E; Zovko, V; Abaffy, T; Barbaric, V; Rudez, I; Jelic, I
MLA Citation
Topic, E, Zovko, V, Abaffy, T, Barbaric, V, Rudez, I, and Jelic, I. "Troponin T and serum enzymes in monitoring open heart surgery." Acta Clinica Croatica 33.1-2 (January 1, 1994): 77-85.
Source
scopus
Published In
Acta clinica Croatica
Volume
33
Issue
1-2
Publish Date
1994
Start Page
77
End Page
85
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