Synthesis and biological evaluation of chalcone derivatives as neuroprotective agents against glutamate-induced HT22 mouse hippocampal neuronal cell death

Article abstract of DOI:10.1016/j.bmcl.2020.127597

Seventeen chalcone analogues were synthesized from 7-methoxy-3,4-dihydronaphthalen1(2H)-one and various aromatic aldehydes under basic conditions and their therapeutic properties were studied in mouse hippocampal cell line HT-22 against neuronal cell deat

Full text of DOI:10.1016/j.bmcl.2020.127597

Journal Pre-proofs
Synthesis and Biological Evaluation of Chalcone Derivatives as Neuroprotec‐
tive Agents against Glutamate-Induced HT22 Mouse Hippocampal Neuronal
Cell Death
Baskar Selvaraj, Uyen Tran Tu Nguyen, Gyewon Huh, Duc Hung Nguyen, Il-
Kyoon Mok, Heesu Lee, Kyungsu Kang, Ae Nim Bae, Dae Won Kim, Jae
Wook Lee
PII:
S0960-894X(20)30708-3
DOI:
Reference:
https://doi.org/10.1016/j.bmcl.2020.127597
BMCL 127597
To appear in:
Bioorganic & Medicinal Chemistry Letters
Received Date:
Revised Date:
Accepted Date:
17 July 2020
22 September 2020
28 September 2020
Please cite this article as: Selvaraj, B., Tran Tu Nguyen, U., Huh, G., Hung Nguyen, D., Mok, I-K., Lee, H.,
Kang, K., Nim Bae, A., Won Kim, D., Wook Lee, J., Synthesis and Biological Evaluation of Chalcone
Derivatives as Neuroprotective Agents against Glutamate-Induced HT22 Mouse Hippocampal Neuronal Cell
Death, Bioorganic & Medicinal Chemistry Letters (2020), doi: https://doi.org/10.1016/j.bmcl.2020.127597
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Synthesis and biological evaluation of
chalcone derivatives as neuroprotective
agents against glutamate-induced HT22
mouse hippocampal neuronal cell death
Leave this area blank for abstract info.
Baskar Selvaraj, Uyen Tran Tu Nguyen, Gyewon Huh, Duc Hung Nguyen, Il-Kyoon Mok, Heesu Lee,
Kyungsu Kang, Ae Nim Bae, Dae Won Kim, Jae Wook Lee
Glutamate
O
[Ca2+
]
O
ROS
NH
C01
MAPKs activation
Cell death

Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com
Synthesis and Biological Evaluation of Chalcone Derivatives as Neuroprotective
Agents against Glutamate-Induced HT22 Mouse Hippocampal Neuronal Cell
Death
Baskar Selvaraja,b,h, Uyen Tran Tu Nguyenc, Gyewon Huha,h, Duc Hung Nguyend, Il-Kyoon Moke, Heesu
Leef, Kyungsu Kangc,h, Ae Nim Baeb,h, Dae Won Kimg, and Jae Wook Leea,b,h,
aNatural Product Research Center, Korea Institute of Science and Technology (KIST), Gangneung, 25451, Republic of Korea
bConvergent Research Center for Diagnosis, Treatment, and Care system of Dementia, Korea Institute of Science and Technology (KIST), Seoul, 02792,
Republic of Korea
cNatural Product Informatics Research Center, Korea Institute of Science and Technology (KIST), Gangneung, 25451, Republic of Korea
dDepartment of Biotechnology, Vietnam-Korea Institute of Science and Technology (VKIST), Hanoi, Vietnam
eInstitute of Food industrialization, Institutes of Green Bio Science & Technology and Graduate School of International Agricultural Technology, Seoul National
University, Pyeongchang-gun, Gangwon-do, 25354, Republic of Korea
fDepartment of Anatomy, College of Dentistry, Gangnueng Wonju National University (GWNU), Gangneung, 25457, Republic of Korea
gDepartment of Biochemistry and Molecular Biology, College of Dentistry, Gangneung Wonju National University (GWNU), Gangneung, 25457, Republic of
Korea
hDivision of Bio-Medical Science and Technology, University of Science and Technology (UST), Daejun, 34113, Republic of Korea
ARTICLE INFO
ABSTRACT
Article history:
Received
Revised
Accepted
Available online
Seventeen chalcone analogues were synthesized from 7-methoxy-3,4-dihydronaphthalen1(2H)-
one and various aromatic aldehydes under basic conditions and their therapeutic properties were
studied in mouse hippocampal cell line HT-22 against neuronal cell death induced by glutamate.
From this study, we selected an analogue C01 as a active compound which showed significantly
high neuroprotection. This compound inhibited Ca2+ influx and reactive oxygen species (ROS)
accumulation inside cells. The glutamate-induced cell death was analyzed by flow cytometry and
it showed that C01 significantly reduced apoptotic or dead cell induced by 5 mM glutamate.
Western blot analysis indicates that glutamate-mediated activation of MAPKs were inhibited by
compound C01 treatment. In addition, the C01enhanced Bcl-2 and decreased Bax, the anti and
pro apoptotic proteins respectively. Further analysis showed that, C01 prevented the nuclear
translocation of AIF (apoptosis inducing factor) and inhibited neuronal cell death. Taken together,
compound C01 treatment resulted in decreased neurotoxicity induced by 5 mM of glutamate. Our
finding confirmed that compound C01 has neuro-therapeutic potential against glutamate-mediated
neurotoxicity.
Keywords:
HT22 cells
MAPK
AIF
ROS
Neuroprotection
2009 Elsevier Ltd. All rights reserved.
———
 Corresponding author. Tel.: +82-33-640-2229; fax: +82-33-642-6410; e-mail: kimdw@gwnu.ac.kr
 Corresponding author. Tel.: +82-33-650-3514; fax: +82-33-650-3529; e-mail: jwlee5@kist.re.kr

Oxidative stress is regarded as a main cause of neuronal
cytotoxicity in the central nervous system (CNS). The neuronal
cells are very sensitive to oxidative stress due to their demand for
relatively high oxidative energy metabolism.1 Glutamate is known
as an excitatory neurotransmitter in the CNS. However, if the
concentration of glutamate is elevated, it can lead to neuronal cell
death due to excessive oxidative stress.2,3,4 Oxidative stress is
implicated in various neuropathologies including Alzheimer’s
disease5, Parkinson’s disease6, brain ischemic injury7, and
amyotrophic lateral sclerosis8. Glutamate-induced oxidative stress
is caused by blocking cysteine uptake, which results in diminished
the level of glutathione (GST) and accumulation of the cellular
reactive oxygen species (ROS).9,10,11 The homeostatic maintenance
of the intracellular calcium ion (Ca2+) levels is crucial in the
physiology and pathology of neuronal cell functions. Under
glutamate-induced oxidative stress, calcium ion (Ca2+) levels are
dramatically increased. Disrupting calcium homeostasis results in
neuronal cell death.10,12,13 Oxidative stress and Ca2+ influx induced
by glutamate activate various cellular signaling pathways in
neuronal cells. The oxidative stress induced by glutamate activate
the mitogen-activated protein kinase (MAPK) pathway proteins
such as ERK and p38 and JNK.14 The MAPK signaling pathway
has critical roles in many cell types in controlling cell death and
survival. In previous reports, ERK activation induced by
glutamate-induced oxidative stress has been shown to be related to
neuronal cell toxicity.15,16 Oxidative stress seems to induce and
reduce the pro and anti-apoptotic proteins respectively.17
Furthermore, apoptosis inducing factor (AIF) activated and
translocate to the nucleus and causes apoptotic cell death.18,19
Accordingly, we assumed that various chalcone derivatives
may have significant neuroprotective potential. Therefore, we
designed and synthesized chalcone derivatives, (E)-2-
benzylidene-7-methoxy-3,4-dihydronaphthalen-1(2H)-one (2), as
novel neuroprotective agents. For the synthesis of (E)-2-
benzylidene-7-methoxy-3,4-dihydronaphthalen-1(2H)-one
derivatives, 7-methoxy-3,4-dihydronaphthalen-1(2H)-one and
various aryl aldehydes were used under basic conditions.
Compared with a conventional chalcone structure (1), (E)-2-
benzylidene-7-methoxy-3,4-dihydronaphthalen-1(2H)-one (2) has
additional fused cyclohexanone ring, which decreases the
flexibility of ,-unsaturated bond of the chalcone scaffold (Fig.
1).
Figure 2. Synthesis of chalcone derivatives and their structures.
O
O
O
R2
R1
1
2
Figure 1. Design of chalcone derivatives.
Chalcones are natural products that possess diverse therapeutic
capabilities such as anti-bacterial, anti-inflammatory and anti-
cancer, among other.20 In previous reports, hydroxyl chalcone and
its derivatives were shown to rescue the neuronal cells from cell
death caused by high concentration of glutamate treatment.21
O
O
O
O
10% NaOH
EtOH
R2
O
3
4
O
O
O
O
O
O
O
O
O
NH
OH
N
O
N
C01
O
C02
O
C03
O
O
O
Cl
O
O
C05
O
C04
O
C06
O
O
O
Br
C08
O
C07
O
C09
O
O
Cl
O
O
O
O
O
O
O
O
Cl
C10
O
C11
O
C12
O
O
Br
O
O
O
O
O
O
O
C15
C14
O
C13
O
O
O
OH
OH
C16
C17

Figure 3. Neuroprotective effects of chalcone derivatives. The neuroprotective
effects of 17 chalcone derivatives were evaluated on HT22 cells treated with 5
mM glutamate. ***P ≤ 0.001 , **P ≤ 0.01 , *P ≤ 0.05
Figure 4. (A) Cell Viability assay to identify live and dead cells revealed that
C01 shows neuroprotective effect. Calcein AM and propidium iodide staining
of live and dead cells (Scale bars, 200 μm). (B) The co-treatment of HT22 cells
with 12.8 μM C01 and 5 mM glutamate (glu) showed more than 80% cell
survival. ∗∗P≤0.01 than the glutamate. (C,D,E,F) FACS analysis of HT22 cells
after staining PI/annexin V. (C) Control. (D) 5 mM glutamate (glu) treatment.
The ratio of apoptotic or dead cells is 84.97% in 5 mM glutamate (glu) treated
cells, (E) but it was decreased to 8.63% by co-treatment of C01 (12.8 μM). (F)
C01 treatment only. The treatment of C01 shows no toxicity in HT22 cells.
The synthesis of chalcone derivatives was performed under
basic conditions using 10% aq. NaOH in EtOH (Fig. 2). These 17
derivatives were initially characterized by liquid chromatography-
mass spectrometry (LC-MS) and further by 1H-NMR. Their
neuroprotection capacity was studied in HT22 using glutamate.
Briefly, the cells were treated with derivatives and 5mM glutamate
for 12hrs time and cell viability was measured by counting the live
cell stained by calcein AM dye and propedium iodide (PI) which
selectively stains dead cells. The 12hr glutamate treatment alone
reduced the cell viability significantly to 50%. However, co-
treatment of the cells with chalcone derivaitves at the
concentration ranging from 4.27μM to 38.46 μM rescued cell
viability to more than 80% (Fig. 3)
The glutamate seems to induce cell death via apoptosis and also
necrosis in a different stages of cell death. The flow cytometry
analysis using the Annexin-V/PI stains showed that 24 h of
glutamate treatment induced apoptotic cell death extensively and
reduced the percentage of live cell to 15.03%. In the presence of
C01, the live cell population was restored to 91.37%. C01
treatment itself showed no cytotoxic effect on HT22 cells (Fig. 4C,
4D, 4E, 4F). This analysis indicates that C01 rescued the HT22
cells from apoptotic or death cells induced by glutamate
neurotoxicity.
Among the 17 synthesized chalcone derivatives, compound
C01 at 12.8 μM exhibited the most potent protective effects against
glutamate-induced cell death (Fig 3A, 3B). In the neuroprotective
assay, compounds C03, C07, C08, C12, and C14, which have a
methoxy group in the phenyl ring showed potent neuroprotective
effects. However, chalcone derivatives C13 and C15, which have
a tri-methoxy group on phenyl ring show weak neuroprotective
effects. Chalcone derivatives C16 and C17 with hydroxyl groups
on the phenyl ring showed no potent neuroprotective effects
against glutamate-induced cytotoxicity.
Glutamate initially blocks cysteine uptake, which results in
depletion of endogenous antioxidant, glutathione, in HT22 cells.
Therefore, antioxidants may show neuroprotective effects by
reducing cellular concentrations of ROS. We therefore tested the
antioxidant effect of C01 using an in-vitro 1, 1-diphenyl-2-
picrylhydrazyl (DPPH) assay. It revealed that C01 has no free
radical scavenging effect (Fig. 1S).
Figure 5. Effect C01 on glutamate-induced elevation of intracellular Ca2+
level and reactive oxygen species. (A) Fluorescent image of ROS sensing using
DCFDA. (B) Fluorescent image of ROS sensor presented as bar graph. (C)

Fluorescent image of Fluo-3 Ca2+ sensor (Scale bars, 200 μm). (D) Fluorescent
image of calcium sensor presented as bar graph. **P ≤ 0.006, *P ≤ 0.03
compared with the glutamate-treated value, Scale bars, 200 μm.
expected the C01 treatment has decreased Bax and increased Bcl-
2 protein levels in the HT22 cells (Fig. 6B).
The previous literatures data shows that glutamate induce the
cell death via caspase independent AIF apoptotic pathway. So, we
studied the C01 effect on AIF translocation to the nucleus by
immunoblot and immunostaining (Fig. 7). Our both results have
showed AIF translocation to the nucleus by the glutamate
treatment. But, the C01 has inhibited the AIF translocation to the
nucleus which thereby prevented the apoptotic cell death in the
HT22 cells (Fig. 7A, B).
The excessive glutamate level elevates the intracellular ROS in
the neuronal cells. To study the effect of C01 on intracellular ROS
generation in HT22 cells we used ROS specific fluorescent dye 2’
,7’-dichlorodihydrofluorescein diacetate (DCF-DA), and we
observed that 12.8 μM C01 has significantly attenuated the ROS
generation induced by glutamate (Fig. 5A, 5B and 2S).
Ca2+ homeostasis is affected by the oxidative stress induced by
glutamate. Many literatures indicate that glutamate-induced
oxidative stress increases cellular Ca2+ uptake, which results in the
activation of apoptosis. Ca2+ fluorescent sensor Fluo-3 AM was
used to measure cellular Ca2+ levels. In this experiment, glutamate
increased Fluo-3 AM fluorescent intensity in HT22 cells.
However, co-treatment of C01 reduced the fluorescent intensity,
indicating that C01 inhibited glutamate-induced cellular Ca2+
uptake in the HT22 cells (Fig. 5C and 5D).
To evaluate the in vivo efficacy of C01, we tested C01 using 6-
hydroxyl
dopamine
(6-OHDA)-treated
C.
elegans
(Caenorhabditis elegans) model, which mimics oxidative stress-
induced dopaminergic neurodegeneration.22 It is known that 6-
OHDA can disrupt dopamine signaling and affect the locomotory
behavior of C. elegans, such as body bending activity.23
When C. elegans were treated with only 6-OHDA, the number
of body movement of the worms was significantly reduced than
the vehicle control group (P < 0.001). Cotreatment of the C.
elegans with C01 (100 µM) significantly increased the number of
body movement compared with that of worms treatment of 6-
OHDA (P < 0.001). These data demonstrated that C01
significantly reduced the neuronal dysfunction mediated by 6-
OHDA in the C. elegans model (Fig. 8).
Here, we have analyzed the neuroprotective effect of the
synthesized chalcone derivatives and found that C01 effectively
protected the HT22 neuronal cell death induced by glutamate.
Also, C01 treatment successfully reduced ROS accumulation and
cellular Ca2+ influx of glutamate. Additionally, it regulates the
expression levels of pro and anti-apoptotic proteins Bcl-2 and Bax.
In the downstream, C01 control the activation of MAP kinase
pathway proteins ERK, JNK and p38. Also, the C01 prevented
AIF-mediated apoptotic cell death by preventing the AIF nuclear
translocation. Further we studied the effect of C01 on NRF2
antioxidant signaling pathway using ARE-luciferase HepG2 stable
cell line. The C01 treatment has highly increased the luciferase
signal and it indicate that C01 seems to strongly activate NRF2
signaling pathway (Fig. 3S). We also studied the C01 effect on
ferroptotic cell death using erastin. Lately some studies showed
that inhibition of ferroptosis also prevent glutamate neurotoxicity
in rat hippocampal slice cultures. But there was no significant
protective effect of C01 against ferroptosis observed. (Fig. 4S). 24
Figure 6. (A-B) Western blot analysis of p-JNK, JNK, p-ERK, ERK
(p44/42 MAP kinase), p-p38, p38, Bcl-2, Bax, and -actin (control) in HT22
cells treated with indicated concentration of C01 and 5 mM glutamate.
This study clearly demonstrates the neuroprotective effect of
C01 against glutamate neurotoxicity which mainly induced by the
elevation of glutamate-induced oxidative stress. This compound
seems to strongly influence the anti-oxidant signaling pathways in
the downstream and attenuate the oxidative stress in the neuronal
cells. As, the oxidative stress is the main cause behind many
neurodegenerative diseases, the C01 might hold novel therapeutic
capabilities and it has to be evaluated further.
The MAPKs pathway associated with neuronal cell death in the
downstream of oxidative stress elevated by glutamate. The
increased intracellular Ca2+ and ROS levels have been suggested
to activate MAPK signaling pathway, including p38, ERK1/2, and
JNK kinases. Recent studies have indicated that inhibition of
MAPK phosphorylation in HT22 cells attenuate glutamate
neurotoxicity. Therefore, we investigated the effect of C01 on
MAPK activation by western blot analysis. It clearly shows that
the glutamate treatment increased activation of MAP kinases
whereas C01 co-treatment has reduced the amount of
phosphorylation of each MAPKs in concentration dependent
manner. (Fig. 6A). Further, the glutamate treatment seems to affect
the expression of pro and anti-apoptotic protein levels such as Bcl-
2 and Bax. To study the anti-apoptotic effect of C01, we also
evaluated Bax and Bcl-2 protein levels by immunoblot. As,

Figure 7. Effect of C01 on AIF nuclear translocation. (A) Western blot
analysis shows that 5 mM of glutamate treatment induced translocation of AIF
into nucleus. The treatment of 12.8 μM of C01 inhibited translocation of AIF
into nucleus. (B) Immunostaining of AIF in HT22 cells showed that 12.8 M
C01 treatment inhibited nuclear translocation of AIF. (i-iv) enlarged image of
the boxed region.
Figure 8. Effect of C01 on 6-OHDA-induced body bending depression in C.
elegans. Age-synchronized N2 adult worms were treated with 40 mM of L-
ascorbic acid, 50 mM of 6-OHDA, and 100 µM of C01 for 1 h. Then, the
number of body bending were counted for 20 s. The values shown are the
meansꢀ±ꢀSD (nꢀ=ꢀ14). The graph is a representative from three independent
experiments. ***Pꢀ<ꢀ0.001, compared to the control. ###P <ꢀ0.001, compared to
the 6-OHDA single-treatment group.

14. Ray PD, Huang BW, Tsuji Y. Reactive oxygen species (ROS)
homeostasis and redox regulation in cellular signaling. Cell
Signal, 2012, 24, 981-990.
15. Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M,
Berman K, Cobb MH. Mitogen-activated protein (MAP) kinase
pathways: regulation and physiological functions. Endocr Rev,
2001, 22, 153-183.
16. Tibbles LA, Woodgett JR. The stress-activated protein kinase
pathways. Cell Mol Lif Sci, 1999, 55, 1230-1254.
17. Kowaltowski AJ, Fiskum G, Redox mechanisms of cytoprotection
by Bcl-2. Antioxid Redox Signal., 2005, 7, 508-514.
Acknowledgments
This work was funded in part by grants from the KIST
Institutional Program (2Z06260, 2Z06440), the National Research
Council of Science & Technology (CRC-15-04-KIST), and the
Ministry of Oceans and Fisheries, Korea (Grant No. 20170488).
18. Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE, Brothers
GM, Mangion J, Jacotot E, Costantini P, Loeffler M, et al.
Molecular characterization of mitochondrial apoptosis-inducing
factor. Nature, 1999, 397, 441-446.
19. Daugas E, Susin SA, Zamzami N, Ferri KE, Irinopoulou T,
Larochette N, Prévost ME, Leber B, Andrews D, Penninger J, et
al. Mitochondrio-nuclear translocation of AIF in apoptosis and
necrosis. FASEB J, 2000, 14, 729-739.
References and notes
1.
2.
Coyle TJ, Puttfarcken P. Oxidative stress, glutamate, and
neurodegerative disorders. Science, 1996, 262, 689-695.
Reynolds IJ, Hastings TG. Glutamate induces the production of
reactive oxygen species in cultured forebrain neurons following
NMDA receptor activation. J. Neurosci, 1995, 15, 3318-3327.
D.W. Choi, Glutamate neurotoxicity and diseases of the nervous
system. Neuron, 1988, 1, 623-634.
Dong XX, Wang Y, Qin ZH. Molecular mechanisms of
excitotoxicity and their relevance to pathogenesis of
neurodegenerative diseases. Acta Pharmacol Sin, 2009, 30, 379-
387.
3.
4.
20. Rani A, Anand A, Kumar K, Kumar V. Recent developments in
biological aspects of chalcones: the odyssey continues. Expert
Opin. Drug Discov., 2019, 14, 249-288.
21. Selvaraj B, Kim DW, Huh G, Lee H, Kang K, Lee JW. Synthesis
and biological evaluation of isoliquiritigenin derivatives as a
neuroprotective agent against glutamate mediated neurotoxicity in
HT22 cells. Bioorg. Med. Chem. Lett. 2020, 30, 127058.
22. Offenburger S-L, Gatner A. 6-hydroxydopamine (6-OHDA)
oxidative stress assay for observing dopaminergic neuron loss in
Caenorhabditis elegans. Bio Protoc. 2018, 8, e3025.
23. Li H, Shi R, Ding F, et al. Astragalus polysaccharide suppresses 6-
hydroxydopamine-induced neurotoxicity in Caenorhabditis
elegans. Oxid Med Cell Longev. 2016;2016:4856761.
doi:10.1155/2016/4856761.
5.
Niikura T, Tajima H, Kita Y. Neuronal cell death in Alzheimer’s
disease and a neuroprotective factor, humanin. Curr
Neuropharmacol, 2006, 4, 139-147.
6.
7.
Cookson MR. α-Synuclein and neuronal cell death. Mol
Neurodegener, 2009, 4, 9.
Kishimoto M, Suenaga J, Takase H, Araki K, Yao T, Fujimura T,
Murayama K, Okumura K, Ueno R, Shimizu N, Kawahara N,
Yamamoto T, Seko Y. Oxidative stress-responsive apoptosis
inducing protein (ORAIP) plays a critical role in cerebral
ischemia/reperfusion Injury. Sci. Rep. 2019, 9, 13512.
Barber SC, Mead RJ, Shaw PJ. Oxidative stress in ALS: A
mechanism of neurodegeneration and a therapeutic target.
Biochem Biophys Acta., 2006, 1762, 1051-1067.
Murphy TH, Miyamoto M, Sastre A, Schnaar RL, Coyle JT.
Glutamate toxicity in a neuronal cell line involves inhibition of
cysteine transport leading to oxidative stress. Neuron, 1989, 2,
1547-1558.
24. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM,
Gleason CE, Patel DN, Bauer AJ, Cantley AM, Tang SW,
Morrison III B, Stockwell BR. Ferroptosis: an iron-dependent
form of nonapoptotic cell death. Cell 2012, 149, 1060-1072.
8.
9.
Supplementary Material
10. Tan S, Schubert D, Maher P. Oxytosis: a novel form of
programmed cell death. Curr Top Med Chem, 2001, 1, 497-506.
11. Kang Y, Tiziani S, Park G, Kaul M, Paternostro G. Cellular
protection using Flt3 and PI3Kα inhibitors demonstrates multiple
mechanisms of oxidative glutamate toxicity. Nat Comm., 2014, 5,
1-12.
12. Tobaben S, Grohm J, Seiler A, Conrad M, Plesnila N, Culmsee C.
Bid-mediated mitochondrial damage is a key mechanism in
glutamate-induced oxidative stress and AIF-dependent cell death
in immortalized HT-22 hippocampal neurons. Cell Death Differ,
2011, 18, 282-292.
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13. Culmsee C, Plesnila N. Targeting Bid to prevent programmed cell
death in neurons. Biochem Soc Trans, 2006, 34, 1334-1340.