Klaus Heese | Hanyang University (original) (raw)

Papers by Klaus Heese

Neuroscience & Biobehavioral Reviews, 2024

Microglia, the intrinsic neuroimmune cells residing in the central nervous system (CNS), exert a ... more Microglia, the intrinsic neuroimmune cells residing in the central nervous system (CNS), exert a pivotal influence on brain development, homeostasis, and functionality, encompassing critical roles during both aging and pathological states. Recent advancements in comprehending brain plasticity and functions have spotlighted conspicuous variances between male and female brains, notably in neurogenesis, neuronal myelination, axon fasciculation, and synaptogenesis. Nevertheless, the precise impact of microglia on sex-specific brain cell plasticity, sculpting diverse neural network architectures and circuits, remains largely unexplored. This article seeks to unravel the present understanding of microglial involvement in brain development, plasticity, and function, with a specific emphasis on microglial signaling in brain sex polymorphism. Commencing with an overview of microglia in the CNS and their associated signaling cascades, we subsequently probe recent revelations regarding molecular signaling by microglia in sex-dependent brain developmental plasticity, functions, and diseases. Notably, C-X3-C motif chemokine receptor 1 (CX3CR1), triggering receptors expressed on myeloid cells 2 (TREM2), calcium (Ca2+), and apolipoprotein E (APOE) emerge as molecular candidates significantly contributing to sex-dependent brain development and plasticity. In conclusion, we address burgeoning inquiries surrounding microglia's pivotal role in the functional diversity of developing and aging brains, contemplating their potential implications for gender-tailored therapeutic strategies in neurodegenerative diseases.

Neurochemistry International, 2024

Family with sequence similarity 72 (FAM72) is a protein-coding gene family located on chromosome ... more Family with sequence similarity 72 (FAM72) is a protein-coding gene family located on chromosome 1 in humans, uniquely featuring four paralogs: FAM72A, FAM72B, FAM72C, and FAM72D. While FAM72's presence as a gene pair with the SLIT-ROBO Rho GTPase-activating protein 2 (SRGAP2) is intriguing, its functional roles, particularly in neural stem cells, remain incompletely understood. This review explores the distinct characteristics of FAM72, shedding light on its expression patterns, potential roles in cell cycle regulation, stem cell renewal and implications in neurogenesis and tumorigenesis.

Experimental Brain Research, Feb 9, 1998

Journal of Molecular Neuroscience, 2004

Molecular Brain Research, Aug 1, 2002

Experimental and Molecular Medicine, Sep 6, 2013

Folia Pharmacologica Japonica, Sep 1, 2004

Molecular Neurobiology, May 16, 2017

PubMed, 2004

Alzheimer's disease (AD) is one of the most common and challenging neurodegenerative diseases in ... more Alzheimer's disease (AD) is one of the most common and challenging neurodegenerative diseases in humans and is characterized by: progressive impairment in cognitive function, degeneration of cholinergic neurons of the basal forebrain (CBF), neurofibrillary tangles and amyloid beta-peptide (Abeta) depositions. The amyloid precursor protein (APP) is a transmembrane protein of which abnormal processing produces Abeta that is associated with the pathogenesis of AD. Neurotrophic factors have attracted much attention for their potential as a remedy for neurological disorders. In this regard, nerve growth factor (NGF) has generated a great interest as a potential target for the treatment of AD. This interest is based on the observation that CBF neurons, which provide the major source of cholinergic innervation to the cerebral cortex and hippocampus, undergo selective and severe degeneration in advanced AD and that the survival of CBF neurons depends upon NGF and its receptors, namely, trkA and p75NTR. This review focuses on recent findings about APP, NGF and their potential signaling-connections to the protein encoded by the 'Sunday-driver' (SYD) gene.

PubMed, Feb 1, 2006

In the mammalian brain, four neurotrophins have been identified: nerve growth factor (NGF), brain... more In the mammalian brain, four neurotrophins have been identified: nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5). NGF exerts an important role in the development and functions of the central and peripheral nervous system. However, it has recently been documented that several types of immune cells, such as mast cells, lymphocytes, basophils and eosinophils, produce, store and release NGF. Accumulating preclinical and clinical data indicate that dysfunctions of NGF and the other neurotrophins may contribute to impaired immune responses and concentration of NGF frequently correlates with disease severity. Thus, the aim of this study was to elucidate the potential signaling mechanisms of cytokine- neurotrophins interactions contributing to increased NGF levels. Our data show that the transcription factor NF-kappaB plays a pivotal role in regulating B-cell-derived NGF expression.

PubMed, 2012

Tianma (Gastrodia elata Blume) is a traditional Chinese medicine (TCM) often used for the treatme... more Tianma (Gastrodia elata Blume) is a traditional Chinese medicine (TCM) often used for the treatment of headache, convulsions, hypertension and neurodegenerative diseases. Tianma also modulates the cleavage of the amyloid precursor protein App and cognitive functions in mice. The neuronal actions of tianma thus led us to investigate its specific effects on neuronal signalling. Accordingly, this pilot study was designed to examine the effects of tianma on the proteome metabolism in differentiated mouse neuronal N2a cells using an iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomics research approach. We identified 2178 proteins, out of which 74 were found to be altered upon tianma treatment in differentiated mouse neuronal N2a cells. Based on the observed data obtained, we hypothesize that tianma could promote neuro-regenerative processes by inhibiting stress-related proteins and mobilizing neuroprotective genes such as Nxn, Dbnl, Mobkl3, Clic4, Mki67 and Bax with various regenerative modalities and capacities related to neuro-synaptic plasticity.

Brain Research, Dec 1, 2000

Molecular Neurobiology, Jan 7, 2015

Neuroscience Letters, Dec 1, 1997

Neuroscience Letters, Aug 1, 1997

Journal of Neurochemistry, Nov 14, 2002

Molecular and Cellular Biochemistry, Dec 14, 2010

Neuroscience & Biobehavioral Reviews, 2024

Neurochemistry International, 2024

Experimental Brain Research, Feb 9, 1998

Journal of Molecular Neuroscience, 2004

Molecular Brain Research, Aug 1, 2002

Experimental and Molecular Medicine, Sep 6, 2013

Folia Pharmacologica Japonica, Sep 1, 2004

Molecular Neurobiology, May 16, 2017

PubMed, 2004

PubMed, Feb 1, 2006

PubMed, 2012

Brain Research, Dec 1, 2000

Molecular Neurobiology, Jan 7, 2015

Neuroscience Letters, Dec 1, 1997

Neuroscience Letters, Aug 1, 1997

Journal of Neurochemistry, Nov 14, 2002

Molecular and Cellular Biochemistry, Dec 14, 2010

Recent data show that as populations age, the number of people affected by neurodegenerative deme... more Recent data show that as populations age, the number of people affected by neurodegenerative dementia is growing at an epidemic pace in various regions of the world. This cross-cultural study examined the relationships among age, gender, ethnicity, religion, and education as well as the attitudes and perceptions related to ageing and dementia. A random sample of 980 participants was selected to represent the multicultural population of Singapore. Data were collected using standardised questionnaires through online portals and by conducting interviews. These data were ultimately analysed by comparing percentage responses and correlation coefficients and by conducting a multiple regression analysis. The results indicate that the perceptions and attitudes of individuals toward ageing and dementia differ among different age groups. Moreover, the level of education attained was significantly correlated with understanding dementia; regardless of education level, Christians had the most positive mindset toward dementia, although most religious individuals did not believe in divine healing. In this study, it was determined that attitudes and perceptions about ageing and dementia are influenced by multiple factors, such as education, age, and religion, and that it is imperative that younger generations develop coping strategies, including healthy lifestyles and social and/or religious communities to provide quality care to the elderly, in general, and to dementia patients, in particular.

The p60 transcription regulator protein (p60TRP) is a basic helix-loop-helix (bHLH) domain-contai... more The p60 transcription regulator protein (p60TRP) is a basic helix-loop-helix (bHLH) domain-containing neuroprotective protein and a member of the G-protein-coupled
receptor (GPCR)-associated sorting protein (GPRASP) family. In the present study, multiple theoretical physico-chemical methods (e.g. Modeller v.9.13, I-TASSER, PROCHECK and ClusPro v2.0 with PIPER) were applied to unveil the three dimensional (3D) protein structure of the p60TRP homodimer protein and explore potential ligand-protein interactions. Our results suggest a Mg2+-containing 3D p60TRP dimer protein that potentially interacts with 5-(1-aziridinyl)-2,4-dinitrobenzamide (CB1954) and [2-(3-dodecylimidazolidin-1-yl)-1-phosphonoethyl] phosphonic acid (B73). The discovery of CB1954 and B73 may serve as a potential lead for further drug screening tests to normalize the p60TRP signaling pathway in neurodegenerative diseases. Interference with p60TRP signaling via CB1954/B73-related molecules might be a novel option for modifying neurodegenerative signaling pathways (e.g. RIN1, PP2A, RanBP5, CREB and SYNJ1) to treat various brain diseases.

p60 transcription regulator protein (p60TRP) facilitates the processing of the amyloid precursor ... more p60 transcription regulator protein (p60TRP) facilitates the
processing of the amyloid precursor protein towards the
non-amyloidogenic pathway by inhibiting the -secretase
action. This protein was initially identified to be downregulated
in the temporal lobe of brains from Alzheimer’s disease
patients. p60TRP is one of the G-protein-coupled receptor
(GPCR)-associated proteins which directly influences the signalling
capacity of GPCRs. In the present study, we investigated
the brain-region-specific proteome profile of transgenic
p60TRP mice to gain an insight into the molecular
events mediated by the long-term effect of neuronal p60TRP
overexpression on brain proteome changes and its potential
implication for neuronal functions in the central nervous system.
Using a proteomics research approach based on isobaric
tags for relative and absolute quantitation, we identified
2,025 proteins, whereby 1,735 proteins were quantified,
out of which 56 were found to be significantly altered in the
cortex and/or hippocampus of neuronal transgenic neuronal
p60TRP mice. Our data suggests that in vivo overexpression
of neuronal p60TRP significantly affects cognitive and
neuroprotective capacities.

Alzheimer’s disease (AD) is a complex brain disorder of the limbic system and association cortice... more Alzheimer’s disease (AD) is a complex brain
disorder of the limbic system and association cortices. The
disease is characterized by the production and deposition of
the amyloid β-peptide (Aβ) in the brain, and the neuropathological
mechanisms involved must be deciphered to gain
further insights into the fundamental aspects of the protein
biology responsible for the development and progression of
this disease. Aβ is generated by the intramembranous cleavage
of the β-amyloid precursor protein, which is mediated
by the proteases β- and γ-secretase. Accumulating evidence
suggests the importance of the coupling of this cleavage
mechanism to G protein signaling. Heterotrimeric G proteins
play pivotal roles as molecular switches in signal
transduction pathways mediated by G protein-coupled
receptors (GPCRs). Extracellular stimuli activate these
receptors, which in turn catalyze guanosine triphosphate–
guanosine diphosphate exchange on the G protein α-
subunit. The activation–deactivation cycles of G proteins
underlie their crucial functions as molecular switches for a
vast array of biological responses. The novel transcription
regulator protein p60 transcription regulator protein and its
related GPCR signaling pathways have recently been described
as potential targets for the development of alternative
strategies for inhibiting the early signaling mechanisms
involved in neurodegenerative diseases such as AD.

In the present study, we show that overexpression of the G-protein-coupled receptor (GPCR)-associ... more In the present study, we show that overexpression of the G-protein-coupled receptor (GPCR)-associated sorting protein p60TRP (transcription
regulator protein) in neural stem cells (NSCs) and in a transgenic mouse model modulates the phosphorylation and proteolytic
processing of amyloid precursor protein (App), N-cadherin (Cdh2), presenilin (Psen) and protein (Mapt). Our results suggest that
p60TRP is an inhibitor of Bace1 (-site App cleaving enzyme) and Psen. We performed several apoptosis assays [Annexin-V, TdT-mediated
dUTP Nick-End Labeling (TUNEL), caspase-3/7] using NSCs and PC12 cells (overexpressing p60TRP and knockdown of p60TRP)
to substantiate the neuroprotective role of p60TRP. Functional analyses, both in vitro and in vivo, revealed that p60TRP promotes
neurosynaptogenesis. Characterization of the cognitive function of p60TRP transgenic mice using the radial arm water maze test demonstrated
that p60TRP improved memory and learning abilities. The improved cognitive functions could be attributed to increased synaptic
connections and plasticity, which was confirmed by the modulation of the -aminobutyric acid receptor system and the elevated expression
of microtubule-associated protein 2, synaptophysin and Slc17a7 (vesicle glutamate transporter, Vglut1), as well as by the inhibition of
Cdh2 cleavage. In conclusion, interference with the p60TRP/ GPCR/secretase signalling pathway might be a new therapeutic target for
the treatment of Alzheimer’s disease (AD).

Heart failure, including myocardial infarction, is the leading cause for death and the incidence ... more Heart failure, including myocardial infarction, is the leading cause for death and the incidence of cardiovascular diseases is predicted to continue to rise worldwide. In the present study we investigated the whole heart proteome profile of transgenic p60- Transcription Regulator Protein (p60TRP) mice to gain an insight into the molecular events caused by the long-term effect of neural p60TRP over-expression on cardiac proteome changes and its potential implication for cardiovascular functions. Using an iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomics research approach, we identified 1148 proteins, out of which 116 were found to be significantly altered in the heart of neural transgenic p60TRP mice. Based on the observed data, we conclude that in vivo neural over-expression of transgenic p60TRP with its neuroprotective therapeutic potential significantly affects cardiovascular capacities.

Active cell death ('apoptosis' or 'programmed cell death') is essential in the development and ho... more Active cell death ('apoptosis' or 'programmed cell death') is essential in the development and homeostasis of multicellular organisms and abnormal inhibition of apoptosis is an indicator of cancer and autoimmune diseases, whereas excessive cell death might be implicated in neurodegenerative disorders such as Alzheimer's disease (AD). Using bioinformatics-, Western-blotting-, yeast-two-hybrid-system-, polymerase chain reaction (PCR)-, and fluorescence microscopy-analyses, we demonstrate here that the neuroprotective protein p60TRP (p60-transcription-regulator-protein) is a basic helix-loop-helix (bHLH) domain-containing member of a new protein family that interacts with the Ran-binding-protein-5 (RanBP5) and the protein-phosphatase-2A (PP2A). The additional findings of its influence on NNT1 and p48ZnF (new-neurotrophin-1, p48-zinc-finger-protein)-signaling and its down-regulation in the brain of AD subjects point to a possible pivotal role of p60TRP in the control of cellular aging and survival.

Apoptosis, the cell's intrinsic death program, plays a crucial role in the regulation of tissue h... more Apoptosis, the cell's intrinsic death program, plays a crucial role in the regulation of tissue homeostasis, and abnormal inhibition of apoptosis is an indicator of cancer and autoimmune diseases, whereas excessive cell death is implicated in neurodegenerative disorders such as Alzheimer's disease (AD). Using cDNA subtraction analysis, we compared p60TRP (p60 transcription regulator protein) expressing cells with control cells during the process of apoptosis and we identified the new zinc-finger protein p48ZnF that is predominantly located in the cytoplasm of the cell. Additionally, we demonstrate here that p48ZnF is up-regulated in rat neuronal PC12 cells upon stimulation with the neurotrophic factor NGF (50 ng/ml). These findings point to a possible pivotal role of p48ZnF in the control of neuronal survival.

p48ZnF is a C3H1 zinc finger domain-containing protein that is involved in the control of gene tr... more p48ZnF is a C3H1 zinc finger domain-containing protein that is involved in the control of gene transcription and translation. In the present study a novel transgenic p48ZnF mouse model is described that is useful for in vivo brain imaging using luciferase as bioluminescence-mediating reporter gene. Yeast two-hybrid screening and western blot analyses revealed Drg1 developmentally regulated GTP binding protein 1) and Pcbp1 (poly (rC)-binding protein 1) as p48ZnF-associated proteins. Interestingly, p48ZnF’ cellular location of action depends on the cell’s differentiation status: nuclear in proliferating cells and
cytoplasmic in differentiated neurons.

Tianma (Gastrodia elata Blume) is a traditional Chinese medicine (TCM) often used for the treatme... more Tianma (Gastrodia elata Blume) is a traditional Chinese medicine (TCM) often used for the treatment of headache, convulsions, hypertension and neurodegenerative diseases. Tianma also modulates the cleavage of the amyloid precursor protein App and cognitive functions in mice. The neuronal actions of tianma thus led us to investigate its specific effects on neuronal signalling. Accordingly, this pilot study was designed to examine the effects of tianma on the proteome metabolism in differentiated mouse neuronal N2a cells using an iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomics research approach. We identified 2178 proteins, out of which 74 were found to be altered upon tianma treatment in differentiated mouse neuronal N2a cells. Based on the observed data obtained, we hypothesize that tianma could promote neuro-regenerative processes by inhibiting stress related proteins and mobilizing neuroprotective genes such as Nxn, Dbnl, Mobkl3, Clic4, Mki67 and Bax with various regenerative modalities and capacities related to neuro-synaptic plasticity.

Gastrodia elata (tianma) is a traditional Chinese herbal medicine (TCM) often used for the treatm... more Gastrodia elata (tianma) is a traditional Chinese herbal medicine (TCM) often used for the treatment of cerebrovascular diseases. In this study, we investigated the effects of tianma on the brain protein metabolism by quantitative proteomics to gain evidence for a direct relationship between tianma treatment and brain functions. One-year-old rats were treated with tianma (~2.5 g/kg/day) for 3 months and the brain tissue proteome was analyzed by using the iTRAQ (isobaric tag for relative and absolute quantification) technology. According to our results, the long-term treatment with tianma could modulate the brain protein metabolism at the proteome level by down-regulating the expressions of various proteins, such as Gnao1 and Dctn2, which are related to neuronal growth cone control and synaptic activities. In addition, tianma treatment also induced the up-regulation of molecular chaperons and proteins related to the misfolded protein response, like Anxa5, and also other proteins involved in Huntington's disease (HD) (e.g. Pacsin1 and Arf3). Concluding, tianma could eventually contribute to activities related to synaptic plasticity and neurorestorative processes and thus might be a novel candidate agent for the treatment of neurodegenerative diseases by regulating the brain proteome.

Tianma is a traditional Chinese medicine (TCM) often used for the treatment of hypertension and h... more Tianma is a traditional Chinese medicine (TCM) often used for the treatment of hypertension and heart diseases.
To elucidate the function of tianma at the molecular level, we investigated the effect of tianma on vascular functions
and aortic protein metabolism. We found that long-term treatment with tianma (~2.5g/kg/day for three months) in
one-year-old rats could enhance acetylcholine (ACh)-induced vasorelaxation in endothelium-intact thoracic aortic rings
against both KCl (80 mM)- and phenylephrine (PE)-induced contraction. By using the iTRAQ (isobaric tag for relative
and absolute quantification) technique, we confirmed from the functional data at the proteome level that tianma treatment
down-regulated the expressions of contractile proteins (e.g. Acta2) and other related structural proteins (e.g. desmin), and
up-regulated the expressions of extracellular matrix (ECM) glycoproteins (e.g. Fbln5) and anti-thrombotic proteins (e.g.
Anxa2) in aortic tissue. By inductive reasoning, tianma could perform its vasodilatory effect not only by inhibiting vascular
smooth muscle contraction, but also by enhancing blood vessel elasticity and stabilizing the arterial structure. Thus,
tianma might become a novel therapeutic herbal medicine for cardiovascular diseases by regulating the aortic proteome
metabolism.

Gastrodia elata (Tianma) is a traditional Chinese medicine often used for the treatment of headac... more Gastrodia elata (Tianma) is a traditional Chinese medicine often used for the treatment of headache, convulsions, hypertension, and cardiovascular diseases. The vasodilatory actions of Tianma led us to investigate its specific effects on memory and learning as well as on Alzheimer's disease (AD)-related signaling. We conducted a radial arm water maze analysis and the novel object recognition test to assess the cognitive functions of Tianma-treated mice. Our data show that Tianma enhances cognitive functions in mice. Further investigations revealed that Tianma enhances the α-secretase-mediated proteolytic processing of the amyloid precursor protein (App) that precludes the amyloid-β peptide production and supports the non-amyloidogenic processing of App which is favorable in AD treatment. We hypothesize that Tianma promotes cognitive functions and neuronal survival by inhibiting β-site App-cleaving enzyme 1 activity and promoting the neuroprotective α-secretase activity.

Gastrodia elata blume (tianma) is a traditional Chinese herb often used in the treatment of convu... more Gastrodia elata blume (tianma) is a traditional Chinese herb
often used in the treatment of convulsions, headaches, and
hypertension. Although interest in neuronal-related actions
of tianma is increasing, minimal studies have been conducted
to determine its specific effects on neuronal cells. This
study was designed to examine the effects of tianma on the
metabolism in differentiated neuroblastoma cells using the
isobaric tag for relative and absolute quantitation (iTRAQ)
technology. Stimulation of these cells with tianma caused
changes in the expression of 38 proteins that were subsequently
classified according to their physiological functions
and association with neurodegenerative diseases. We identified
six proteins with altered functional activities in neurodegenerative
disease states that were modulated by tianma:
triosephosphate isomerase (Tpi1), peptidyl-prolyl cis-trans
isomerase A (Ppia), neural cell adhesion molecule 1 (Ncam1),
ubiquitin carboxyl-terminal hydrolase isozyme L1 (Uchl1),
septin-2 (Sept2) and heat shock protein 90 (Hsp90aa1). We postulate that tianma mediates its neuroprotective effects
via upregulation of Ncam1, Hsp90aa1, Tpi1 and Ppia while
downregulating Sept2 and Uchl1. These changes in protein
expression aid in the restoration of the intracellular environment
to a metabolically balanced state, promoting cell survival.
Based on these observed data, we conclude that tianma
has therapeutic potential, especially for neurodegenerative
diseases.

Tianma (Rhizoma gastrodiae) is the dried rhizome of the plant Gastrodia elata Blume (Orchidaceae ... more Tianma (Rhizoma gastrodiae) is the dried rhizome of the plant Gastrodia elata Blume (Orchidaceae family).
As a medicinal herb in traditional Chinese medicine (TCM) its functions are to control convulsions, pain,
headache, dizziness, vertigo, seizure, epilepsy and others. In addition, tianma is frequently used for the
treatment of neurodegenerative disorders though the mechanism of action is widely unknown. Accordingly,
this study was designed to examine the effects of tianma on the proteome metabolism in differentiated
human neuronal SH-SY5Y cells to explore its specific effects on neuronal signaling pathways. Using
an iTRAQ (isobaric tags for relative and absolute quantitation)-based proteomics research approach, we
identified 2390 modulated proteins, out of which 406 were found to be altered by tianma in differentiated
human neuronal SH-SY5Y cells. Based on the observed data, we hypothesize that tianma promotes
neuro-regenerative signaling cascades by controlling chaperone/proteasomal degradation pathways (e.g.
CALR, FKBP3/4, HSP70/90) and mobilizing neuro-protective genes (such as AIP5) as well as modulating
other proteins (RTN1/4, NCAM, PACSIN2, and PDLIM1/5) with various regenerative modalities and capacities
related to neuro-synaptic plasticity.

Gastrodia elata (Tianma) is a traditional Chinese herb with demonstrated vasodilatory effects. Th... more Gastrodia elata (Tianma) is a traditional Chinese herb with demonstrated vasodilatory effects. This pilot study examined the effects of Tianma treatment on bladder smooth muscle contractility. Rats were treated with 2.5 g Tianma per kg body weight over 7 weeks. Contractility was measured in detrusor strips isolated in both transverse (Tr) and longitudinal (Lg) directions with the urothelium intact (+UE) or denuded (-UE). Spontaneous phasic activity was enhanced in longitudinal +UE strips. No differences between control and Tianma-treated detrusor strips were detected in contractions elicited by K+-Krebs’ solution or carbachol. Isoprenaline (IPNA)-induced relaxation remained unchanged in –UE strips after Tianma treatment. However, potency of IPNA was lower in Tianma-treated +UE strips in the longitudinal direction. These findings provided an initial assessment of how Tianma altered bladder smooth muscle function. It will be of interest in future investigations to elucidate the mechanisms via which Tianma exerts its actions on the bladder.

Stem cells intrigue. They have the ability to divide exponentially, recreate the stem cell compar... more Stem cells intrigue. They have the ability to divide exponentially, recreate the stem cell compartment, as well as create differentiated cells to generate tissues. Therefore, they should be natural candidates to provide a renewable source of cells for transplantation applied in regenerative medicine. Stem cells have the capacity to generate specific tissues or even whole organs like the blood, heart, or bones. A subgroup of stem cells, the neural stem cells (NSCs), is characterized as a self-renewing population that generates neurons and glia of the developing brain. They can be isolated, genetically manipulated and differentiated in vitro and reintroduced into a developing, adult or a pathologically altered central nervous system. NSCs have been considered for use in cell replacement therapies in various neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Characterization of genes with tightly controlled expression patterns during differentiation represents an approach to understanding the regulation of stem cell commitment. The regulation of stem cell biology by the ATP-binding cassette (ABC) transporters has emerged as an important new field of investigation. As a major focus of stem cell research is in the manipulation of cells to enable differentiation into a targeted cell population; in this review, we discuss recent literatures on ABC transporters and stem cells, and propose an integrated view on the role of the ABC transporters, especially ABCA2, ABCA3, ABCB1 and ABCG2, in NSCs' proliferation, differentiation and regulation, along with comparisons to that in hematopoietic and other stem cells.

The protein family of the neurotrophins (NTs) comprises structurally and functionally related mol... more The protein family of the neurotrophins (NTs) comprises
structurally and functionally related molecules such as nerve
growth factor (NGF) which infl uences the proliferation, differentiation,
survival and death of neuronal cells. In addition
to their established functions for cell survival, NTs also mediate
higher brain activities such as learning and memory.
Changes in NT expression levels have thus been implicated
in neurological diseases such as Alzheimer’s disease (AD), an
age-related neurodegenerative disorder that is characterized
by progressive loss of memory and deterioration of
higher cognitive functions. The present review provides an
overview of the functional role of NGF in neural stem cells
and AD while pointing to a potential application of this peptide
for the treatment of AD.

Besides its wide range of action as a proinflammatory cytokine in the immune system, interleukin-... more Besides its wide range of action as a proinflammatory cytokine in the immune system, interleukin-6 (IL-6) has also
attracted much attention due to its influence on the nervous system. In the present study we show that the designer fusion
protein H-IL-6, consisting of IL-6 and its specific receptor IL-6R-, but not IL-6 alone, mediates both neuro- as well as
gliogenesis. Using immunocytochemistry, Western blot, and patch-clamp recording, we demonstrate that H-IL-6 induces
the differentiation of neural stem cells (NSCs) specifically into glutamate-responsive neurons and two morphological
distinctive astroglia cell types. H-IL-6–activated neurogenesis seems to be induced by the MAPK/CREB (mitogenactivated
protein kinase/cAMP response element-binding protein) cascade, whereas gliogenesis is mediated via the
STAT-3 (signal transducers and activators of transcription protein-3) signaling pathway. Our finding that IL-6 mediates
both processes depending on its specific soluble receptor sIL-6R- has implications for the potential treatment of
neurodegenerative diseases.

Neurospheres can be generated from the mouse fetal forebrain by exposing multipotent neural stem ... more Neurospheres can be generated from the mouse fetal forebrain by exposing multipotent neural stem cells
(NSCs) to epidermal growth factor (EGF). In the presence of EGF, NSCs can proliferate continuously while retaining the
potential to differentiate into neurons, astrocytes and oligodendrocytes. We examined the expression pattern of the
neurotrophin (NT) receptors tropomysin-related kinase (TRK)-A, TRK-B, TRK-C and p75 neurotrophin receptor
(p75NTR) in NSCs and the corresponding lineage cells. Furthermore, we analyzed the action of the NT Brain-Derived
Neurotrophic Factor (BDNF) on NSCs’ behavior. The effects of BDNF on NSC proliferation and differentiation were
examined together with the signaling pathways by which BDNF receptors transduce signaling effects. We found that all
the known NT receptors, including the truncated isoforms of TRK-B (t-TRK-B) and TRK-C (t-TRK-C), were expressed
by Nestin-positive cells within the neurosphere. Proliferation was enhanced in Nestin-positive and BrdU-positive cells in
the presence of BDNF. In particular, we show that t-TRK-B was abundantly expressed in NSCs and the corresponding
differentiated glia cells while full length TRK-B (fl-TRK-B) was expressed in fully differentiated post-mitotic neurons
such as the neuronal cells of the newborn mouse cortex, suggesting that BDNF may exert its proliferative effects on NSCs
through the t-TRK-B receptor. Finally, we analyzed the cell fates of NSCs differentiated with BDNF in the absence of
EGF and we demonstrate that BDNF stimulated the formation of differentiated cell types in different proportions through
the MAP kinase, AKT and STAT-3 signaling pathways. Thus, the in-vivo regulation of neurogenesis may be mediated by
the summation of signals from the BDNF receptors, in particular the t-TRK-B receptor, regulating physiological fates as
diverse as normal neural replacement, excessive neural loss, or tumor development.

The effect of neurotrophin-4 (Ntf4) on mouse embryonic (day-14) neural stem cell (mE14-NSC) fate ... more The effect of neurotrophin-4 (Ntf4) on mouse
embryonic (day-14) neural stem cell (mE14-NSC) fate
determination and the mechanisms involved were investigated.
Using primary mE14-NSCs, immunocytochemistry
and molecular-cell biological methods, such as Westernblotting,
we characterized the effect of Ntf4 on mE14-NSC
differentiation. Obtained in-vitro data revealed an interesting
phenomenon of Ntf4 action resulting in enhanced
mE14-NSC commitment to progenitor cells of the neuronal
lineage. During this process, Ntf4 suppresses the interleukin
6 (Il6) family receptor and the Notch signalling pathways
by modulating their specific receptor cleavages. The
observed lineage commitment is controlled via an Ntf4-
mediated modulation of protein kinase B (PKB/Akt)
activity and characterized by a decreased Stat3 (signal
transducer and activator of transcription-3) phosphorylation
status. These findings suggest that the Ntf4-activated
signalling cascade is responsible for initiating a concert
among sheddases, kinases, and phosphatases to mediate
neurogenesis.

The Nerve Growth Factor (NGF) is the prototypic member of the neurotrophin (NT) family, which pla... more The Nerve Growth Factor (NGF) is the prototypic member of the neurotrophin (NT) family, which plays an essential role in the development and functioning of the vertebrate nervous system. Although originally defined by their actions on neuronal survival and differentiation in the peripheral (PNS) and central nervous systems (CNS), accumulating data indicate the presence of extensive interactions between the NTs and the immune system. NTs are released normally during lymphocyte and leukocyte development by the bone marrow and the thymus and later by secondary lymph organs to maintain responsiveness of these circulating naïve and memory immune cells. Functional NT receptors have been detected on the cells of the immune system and increased levels of NGF protein are found during the acute phase of various diseases with a significant inflammatory component. Furthermore, in certain conditions such as allergic asthma, the released NTs exacerbate the severity of the inflammation and prolong the diseased state. However, in the CNS, if one can control homeostasis of the internal environment, then the natural response of the infiltrating immune cells to release these NTs can be used to intervene at key points in the disease progression. These wider functions are likely to be of concern in any attempted therapeutic use of NGF or related NTs.

In the mammalian brain, four neurotrophins have been identified: nerve growth factor (NGF), brain... more In the mammalian brain, four neurotrophins have been identified: nerve growth factor (NGF), brain-derived
neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5). NGF exerts an important role
in the development and functions of the central and peripheral nervous system. However, it has recently been
documented that several types of immune cells, such as mast cells, lymphocytes, basophils and eosinophils, produce,
store and release NGF. Accumulating preclinical and clinical data indicate that dysfunctions of NGF and the other
neurotrophins may contribute to impaired immune responses and concentration of NGF frequently correlates with
disease severity. Thus, the aim of this study was to elucidate the potential signaling mechanisms of cytokineneurotrophins
interactions contributing to increased NGF levels. Our data show that the transcription factor NF-κB plays a pivotal role in regulating B-cell-derived NGF expression.

lzheimer's disease (AD) is characterized by the presence of beta-amyloid (Abeta) protein deposits... more lzheimer's disease (AD) is characterized by the presence of beta-amyloid (Abeta) protein deposits in the brain and increased Abeta (1-42) peptide production is thought to be one of the early events in the pathogenesis of AD that leads to progressive neurodegenerative processes and dementia. Using cDNA subtraction and reverse transcription-polymerase chain reaction, we examined the Abeta (1-42) peptide-induced gene expression in rat neuroblastoma B104 cells. In addition we hypothesized that interleukin-11 (IL-11) supports neuronal survival. We found that Abeta (1-42) activates L-phosphoserine phosphatase in neuronal cells which is inhibited by IL-11. Moreover, IL-11 inhibits Abeta (1-42)-induced neurotoxicity in a dose-dependent manner. Our study suggests that L-phosphoserine phosphatase may play a role in altered neuronal function in AD via enhancing glutamate-induced neurotoxicity by D-serine and the IL-11 receptor system may act as a neuroprotective cytokine in human brain.

There is increasing evidence that nerve growth factor (NGF) acts on cells of the immune system, a... more There is increasing evidence that nerve growth factor (NGF) acts on cells of the immune system, apart from its neurotrophic
effects. In human basophils, NCF potentiates mediator release and primes the cells to produce leukotriene C, in response to C5a. It is, however, unknown whether other homologous neurotrophins also act outside the nervous system, and whether activation of basophils by NCF requires interaction with trk tyrosine kinase receptors, the low affinity NGF receptor (LNCFR), or both. A triple mutant NCF designed to interrupt binding to the LNCFR was found to activate basophils with equal efficacy as wild-type NGF, demonstrating that the LNCFR is not necessary. Despite a 10 times lower potency of mutant NGF, no LNCFR expression was detected by FACS analysis. Brain-derived neurotrophic factor, which interacts with trkB, was inactive at concentrations up to 1000 ng/ml (>30,000-fold lower potency than NGF), while neurotrophin-3, which is thought to interact with trkC, trkB, and more weakly with trk, induced a threshold effect at 300 ng/ml (-10,000-fold lower potency), demonstrating that 1) the LNCFR cannot deliver a direct signal; and 2) basophils do not express functional trkB and trkC receptors. In agreement with the functional data, basophils (in contrast to other granulocyte types) expressed mRNA for trk, but not trkB or trkC, and no or minimal mRNA for LNGFR. These data demonstrate that human blood basophils express functional trk receptors that do not require the participation of LNCFR, and that, among the neurotrophin family, NCF is unique in priming basophils.

Microglial response to stimuli is characterized by secretion of both neurotoxic and neurotrophic ... more Microglial response to stimuli is characterized by secretion of both neurotoxic and neurotrophic factors. Various adenosine receptor agonists stimulated the production of nerve growth factor (NGF) in microglia. Using reverse transcription-polymerase chain reaction (RTPCR)- and ELISA-techniques, we show that the mixed A1- and A2-agonist 5¢-(N-ethylcarboxamido)-adenosine (NECA) induces an increase in NGF mRNA expression and NGF protein release. Whereas the A1-specific agonist cyclopentyladenosine (CPA) only minimally affected NGF release, the A2a-specific agonist CGS-21680 triggered the greatest increase in microglial NGF synthesis. Analyzing the selective antagonist (E)-8-(3,4-dimethoxystyryl)-1,3-dipropyl-7-methylxanthine (EXIP), as well as modulators of the cyclic AMP (cAMP) pathway, we identified an adenosine A2a-receptor-sensitive, cAMP-mediated mechanism of microglial NGF synthesis. Our results indicate that A2aadenosine receptors modulate microglial neurotrophin expression and release.

Microglia/brain macrophages activated in response to injury, infection, or inflammation of the ce... more Microglia/brain macrophages activated in response to injury, infection, or inflammation of the central nervous system (CNS) mediate both neurotoxic and neurotrophic activities. Although the cytotoxic effects of microglia have been analyzed in detail, little is known about the signaling pathways involved in microglial neurotrophin expression. Using purified rat microglial cell cultures, the effects of inflammatory agents such as lipopolysaccharide (LPS) on microglial nerve growth factor (NGF) expression were studied. Application of LPS (0.1–100 ng/ml) induced a rapid (2–4 h), dose-dependent increase in NGF mRNA expression followed by enhanced release of NGF protein within 24 h. To determine whether the transcription factor NF-kB, known to be stimulated in activated microglia, is involved in inflammatory mediator-induced NGF expression, we used the NF-kB inhibitor pyrrolidine dithiocarbamate (PDTC). Addition of PDTC (100 μM) to microglia completely abolished LPS-induced NGF synthesis, suggesting a key role for NF-kB in microglial NGF expression by inflammatory mediators. In conclusion, NF-kB-controlled NGF expression by activated microglia appears to contribute to the cross-talk between the immune and nervous systems during inflammation in the CNS.

Nerve growth factor (NGF) accumulates at sites of inflammation and modulates local immune reactio... more Nerve growth factor (NGF) accumulates at sites of inflammation and modulates local immune reactions. To characterize the mechanisms of cytokine-induced NGF expression under physiological and pathophysiological conditions, we have used cultured glomerular mesangial cells, which play a key role in glomerular inflammatory diseases such as diabetic nephropathy. To study the effects of high glucose on cytokine-induced NGF expression, rat mesangial cells were treated with the cytokines interleukin-1beta and tumor necrosis factor alpha under normal (1.0 g/L) and high (4.5 g/L) glucose concentrations. In the presence of high glucose concentrations, the cytokines drastically potentiated NGF protein but not mRNA expression when compared to physiological glucose levels. The specific protein kinase C inhibitors Ro31-8220 and CGP41251 suppressed cytokine-induced NGF expression. Moreover, blocking the oxidative activation of the protein kinase C pathway by N-acetylcysteine inhibited glucose effects on NGF synthesis. Neutralizing antibodies against transforming growth factor-beta inhibited cytokine-induced NGF expression under normal glucose concentrations but not under high glucose conditions. Enhanced expression of NGF under high glucose conditions may contribute to kidney diseases such as diabetic nephropathy.

Neurotrophins (NTs) such as nerve growth factor (NGF) as well as cytokines, for example, interleu... more Neurotrophins (NTs) such as nerve growth factor (NGF) as well as cytokines, for example, interleukin-6 (IL-6), are communicators between the nervous and immune systems. There is evidence for mutual interactions between NTs and cytokines. Strategies are being developed to elucidate the molecular mechanism/s of interactions and to understand how cytokines are involved in health and disease. Analysis of underlying signaling pathways in glial cells indicates that different transcription factors, such as NF-kB, cAMP–responsive-element binding protein (CREB), and activator protein 1 (AP-1), are involved in NT induction. IL-6 and NTs of the NGF family are coexpressed at sites of nerve injury. Interactions of these factors could modulate both neuronal de- and regeneration: IL-6 in conjunction with its soluble IL-6 receptor induces a specific pattern of NTs in astrocytes in defined brain regions. This indicates that the IL-6 system mediates a local supply of NTs that participate in diverse CNS functions, such as protection of neurons from insults, neuronal survival, and neuroimmune responses.

Inflammatory processes involving reactive microglia appear to contribute to neuronal degeneration... more Inflammatory processes involving reactive microglia appear
to contribute to neuronal degeneration in the central nervous
system (CNS). However, the fact that microglia release
neurotrophic factors led us to investigate the signaling
mechanisms involved in neurotrophin (NT) synthesis in a
human microglial cell line. This cell line shows typical
properties of human microglia including expression of NTs
such as nerve growth factor (NGF), brain-derived
neurotrophic factor (BDNF), NT-3 and NT-4/5, as well as
the NGF-receptor TrkA and the low affinity NT-receptor
p75NTR. We have analyzed various inflammatory stimuli
and their potency in modulating neurotrophin expression in
microglial cells. Exposure of these cells to complement factor
C3a induces NGF, BDNF and NT-3 expression. Analyzing
the mechanisms involved in this induction revealed that, in
contrast to cytokine-stimulated microglial NT expression
which involves the transcription factor NF-kB, other
transcription factors such as AP-1 (activator protein 1) and
CREB (cAMP-response element-binding protein) contribute
to complement-dependent NT synthesis in human microglial
cells. We discuss the potential of complement receptor agonists
as therapy for the treatment of neurodegenerative diseases.

In an aging society with dementia imposing an increasing threat to higher brain cognitive functio... more In an aging society with dementia imposing an increasing threat to higher brain cognitive functions, understanding the molecular and cellular events of adult neurogenesis is imperative. Interleukin-6 (IL-6), along with its agonistic acting soluble receptor sIL-6R (the combined proteins are also known as Hyper-IL-6), is a promising cytokine that can support neurogenesis under conditions of neurodegeneration when neuron replacement is needed. In contrast to the previously reported gliogenic effects of activation of the IL-6–signal transducer and activator of transcription 3 (STAT3) axis, this review summarizes recent studies showing that IL-6 activation can be neurogenic and has potential therapeutic applications for the treatment of neurodegenerative diseases such as Parkinson's disease.

In an aging society with an increasing threat to higher brain cognitive functions due to dementia... more In an aging society with an increasing threat to
higher brain cognitive functions due to dementia, it becomes
imperative to identify new molecular remedies for supporting
adult neurogenesis. Interleukin-6 (IL-6) is a promising cytokine
that can support neurogenesis under conditions of neurodegeneration,
and neuron replacement is eventually possible
due to its agonistic acting soluble receptor sIL-6R. Here, we
report that activation of the IL-6–signal transducer and activator
of transcription 3 (STAT3) axis is neurogenic and has potential
therapeutic applications for the treatment of neurodegenerative
diseases such as Parkinson’s disease (PD).

Expression of interleukin-6 (IL-6) and fibroblast growth factor-2 (FGF-2) in Schwann cells is mod... more Expression of interleukin-6 (IL-6) and fibroblast growth factor-2 (FGF-2) in Schwann cells is modulated by external stimuli. To study possible interactions of both factors we have analyzed mutual effects of exogenous IL-6 and FGF-2 on the expression of each other and the corresponding receptor (R) molecules IL-6R and FGFR1 after peripheral nerve lesion in vivo and in vitro using cultured Schwann cells. Using rat Schwann cells we found that IL-6 did not exert any effects on the expression of FGF-2 and FGF receptor type 1 (R1) whereas exogenously applied 18-kD FGF-2 strongly increased the expression of the mRNAs of IL-6 and its receptor. In addition, immortalized Schwann cells over-expressing the 18-kD FGF-2 isoform showed elevated levels of IL-6 and IL-6R whereas immortalized Schwann cells over-expressing the high-molecular-weight isoforms (21 kD and 23 kD) displayed unaltered IL-6 and IL-6R expression levels. According to in situ hybridization studies of intact and crushed sciatic nerves in vivo, Schwann cells seems to be the main source of IL-6 and IL-6R. Following sciatic nerve crush, the FGF-2 and the IL-6 system are upregulated after the first hours. Furthermore, we showed that the early increase of the FGF-2 protein is mainly confined to the 18-kD isoform. These results are consistent with the idea of a functional coupling of FGF-2 and the IL-6 system in the early reaction of Schwann cells to nerve injury.

Increasing evidence supports an essential role for interleukin-6 (IL-6) in the development, diffe... more Increasing evidence supports an essential role for interleukin-6 (IL-6) in the development, differentiation, as well as de- and re-generation of neurons in the central nervous system (CNS). Both IL-6 and its specific receptor (IL-6R) are expressed on neurons and glial cells including astrocytes. In this study, we have analyzed the responses of primary rat astrocytes of various brain regions to IL-6 with respect to morphological changes and neurotrophin expression. Since IL-6 alone failed to initiate effects on astrocytes, we have examined whether the soluble IL-6R (sIL-6R) can modulate the responsiveness of to IL-6 in these cells. For this purpose, we used a highly active fusion protein of IL-6 and sIL-6R, which is designated Hyper-IL-6 (H-IL-6).We show that treatment of cultured astrocytes with Hyper-IL-6 promotes region-specific morphological changes of GFAP-positive astrocytes from typical stellate- to fibrous-like cells. In addition, we find that Hyper-IL-6 induces expression of neurotrophins (NTs) of the nerve growth factor (NGF)-family in a dose-dependent manner. Interestingly, astrocytes of various brain regions show differing patterns of cytokine-induced NT expression: NGF is maximally induced in cortex and hippocampus, NT-3 in hippocampus, and NT-4/5 in cortex and cerebellum. In summary, our results indicate that IL-6 in conjunction with sIL-6R regulates specific neurotrophin expression in astrocytes in a brain region dependent manner. Thus, the IL-6 system provides a local supply of neurotrophins that participate in diverse CNS functions such as protection of neurons from insults, neuronal survival, and neuro-immune responses.

We quantitated interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R) and soluble form of the IL-6 ... more We quantitated interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R) and soluble form of the IL-6 signal-transducing protein gp130
(sgp130) in cerebrospinal fluid (CSF) of patients with Alzheimer’s disease (AD) (n = 17) and control subjects (n = 18) using
sensitive enzyme-linked immunosorbent assays (ELISA). Our results show that none of the parameters examined was significantly different in CSF of AD patients as compared to control age-matched non-demented patients. We conclude that CSF levels of IL-6 and their soluble receptors do not necessarily reflect local changes of the IL-6 system that has been shown to be involved in neurodegenerative events occurring in AD. Levels of sgp130 are substantially high (approximately 100 ng/ml) in the CSF of all individuals probably representing a high antagonistic potential.

In recent years, there has been an increasing worldwide interest in traditional Chinese medicine ... more In recent years, there has been an increasing worldwide interest in traditional Chinese medicine (TCM). This increasing demand for TCM needs to be accompanied by a deeper understanding of the mechanisms of action of TCM-based therapy. However, TCM is often described as a concept of Chinese philosophy, which is incomprehensible for Western medical society, thereby creating a gap between TCM and Western medicine (WM). In order to meet this challenge, TCM research has applied proteomics technologies for exploring the mechanisms of action of TCM treatment. Proteomics enables TCM researchers to oversee various pathways that are affected by treatment, as well as the dynamics of their interactions with one another. This review discusses the utility of comparative proteomics to better understand how TCM treatment may be used as a complementary therapy for Alzheimer's disease (AD). Additionally, we review the data from comparative AD-related TCM proteomics studies and establish the relevance of the data with available AD hypotheses, most notably regarding the ubiquitin proteasome system (UPS).

One of the shared hallmarks of neurodegenerative diseases is the accumulation of misfolded protei... more One of the shared hallmarks of neurodegenerative
diseases is the accumulation of misfolded proteins. Therefore,
it is suspected that normal proteostasis is crucial for neuronal
survival in the brain and that the malfunction of this mechanism
may be the underlying cause of neurodegenerative diseases.
The accumulation of amyloid plaques (APs) composed
of amyloid-beta peptide (Aβ) aggregates and neurofibrillary
tangles (NFTs) composed of misfolded Tau proteins are the
defining pathological markers of Alzheimer’s disease (AD).
The accumulation of these proteins indicates a faulty protein
quality control in the AD brain. An impaired ubiquitinproteasome
system (UPS) could lead to negative consequences
for protein regulation, including loss of function.
Another pivotal mechanism for the prevention of misfolded
protein accumulation is the utilization of molecular chaperones.
Molecular chaperones, such as heat shock proteins
(HSPs) and FK506-binding proteins (FKBPs), are highly
involved in protein regulation to ensure proper folding and
normal function. In this review, we elaborate on the molecular
basis of AD pathophysiology using recent data, with a particular
focus on the role of the UPS and molecular chaperones as
the defensive mechanism against misfolded proteins that have
prion-like properties. In addition, we propose a rational therapy
approach based on this mechanism.

Amyloid-beta peptide (Abeta) achieves neurodegeneration through unknown mechanisms. To elucidate ... more Amyloid-beta peptide (Abeta) achieves neurodegeneration through unknown mechanisms. To elucidate some of these mechanisms, we conducted a cDNA subtraction analysis of Abeta-mediated neurotoxicity in neuronal cells and observed an up-regulation of the novel gene p17. The p17 protein was also found elevated in Alzheimer's disease (AD) mouse model. Here, we characterised p17 primarily in cell lines with respect to its localisation, function and physiological expression. We discovered that p17 acts downstream of protein kinase C and inhibits the tyrosine receptor kinase B-brain-derived neurotrophic factor (TrkB-BDNF) pathway. It impedes survival factors and enhances amyloid precursor protein expression thus suggesting its involvement in the Abeta-mediated pro-apoptotic pathways in AD.

Alzheimer's disease (AD) is a complex brain disorder of the limbic system and association cortice... more Alzheimer's disease (AD) is a complex brain disorder of the limbic system and association cortices. The disease is characterized by the production and deposition of the amyloid β-peptide (Aβ) in the brain, and the neuropathological mechanisms involved must be deciphered to gain further insights into the fundamental aspects of the protein biology responsible for the development and progression of this disease. Aβ is generated by the intramembranous cleavage of the β-amyloid precursor protein, which is mediated by the proteases β- and γ-secretase. Accumulating evidence suggests the importance of the coupling of this cleavage mechanism to G protein signaling. Heterotrimeric G proteins play pivotal roles as molecular switches in signal transduction pathways mediated by G protein-coupled receptors (GPCRs). Extracellular stimuli activate these receptors, which in turn catalyze guanosine triphosphate-guanosine diphosphate exchange on the G protein α-subunit. The activation-deactivation cycles of G proteins underlie their crucial functions as molecular switches for a vast array of biological responses. The novel transcription regulator protein p60 transcription regulator protein and its related GPCR signaling pathways have recently been described as potential targets for the development of alternative strategies for inhibiting the early signaling mechanisms involved in neurodegenerative diseases such as AD.

In the present study, we show that overexpression of the G-protein-coupled receptor (GPCR)-associ... more In the present study, we show that overexpression of the G-protein-coupled receptor (GPCR)-associated sorting protein p60TRP (transcription regulator protein) in neural stem cells (NSCs) and in a transgenic mouse model modulates the phosphorylation and proteolytic processing of amyloid precursor protein (App), N-cadherin (Cdh2), presenilin (Psen) and τ protein (Mapt). Our results suggest that p60TRP is an inhibitor of Bace1 (β-site App cleaving enzyme) and Psen. We performed several apoptosis assays [Annexin-V, TdT-mediated dUTP Nick-End Labeling (TUNEL), caspase-3/7] using NSCs and PC12 cells (overexpressing p60TRP and knockdown of p60TRP) to substantiate the neuroprotective role of p60TRP. Functional analyses, both in vitro and in vivo, revealed that p60TRP promotes neurosynaptogenesis. Characterization of the cognitive function of p60TRP transgenic mice using the radial arm water maze test demonstrated that p60TRP improved memory and learning abilities. The improved cognitive functions could be attributed to increased synaptic connections and plasticity, which was confirmed by the modulation of the γ-aminobutyric acid receptor system and the elevated expression of microtubule-associated protein 2, synaptophysin and Slc17a7 (vesicle glutamate transporter, Vglut1), as well as by the inhibition of Cdh2 cleavage. In conclusion, interference with the p60TRP/ GPCR/secretase signalling pathway might be a new therapeutic target for the treatment of Alzheimer's disease (AD).

This study is aimed at gaining insights into the brain site-specific proteomic senescence signatu... more This study is aimed at gaining insights into the brain site-specific proteomic senescence signature while comparing physiologically aged brains with aging-related dementia brains (for example, Alzheimer's disease (AD)). Our study of proteomic differences within the hippocampus (Hp), parietal cortex (pCx) and cerebellum (Cb) could provide conceptual insights into the molecular mechanisms involved in aging-related neurodegeneration. Using an isobaric tag for relative and absolute quantitation (iTRAQ)-based two-dimensional liquid chromatography coupled with tandem mass spectrometry (2D-LC-MS/MS) brain site-specific proteomic strategy, we identified 950 proteins in the Hp, pCx and Cb of AD brains. Of these proteins, 31 were significantly altered. Most of the differentially regulated proteins are involved in molecular transport, nervous system development, synaptic plasticity and apoptosis. Particularly, proteins such as Gelsolin (GSN), Tenascin-R (TNR) and AHNAK could potentially act as novel biomarkers of aging-related neurodegeneration. Importantly, our Ingenuity Pathway Analysis (IPA)-based network analysis further revealed ubiquitin C (UBC) as a pivotal protein to interact with diverse AD-associated pathophysiological molecular factors and suggests the reduced ubiquitin proteasome degradation system (UPS) as one of the causative factors of AD.

Background: Alzheimer's disease (AD) is an age-related neurodegenerative disorder that is charact... more Background: Alzheimer's disease (AD) is an age-related neurodegenerative disorder that is characterized by a progressive loss of higher cognitive functions. The brain of an individual with AD exhibits extracellular senile plaques (SPs) of aggregated amyloid-beta peptide (Abeta) and intracellular neurofibrillary tangles (NFTs). Given the critical role of neuronal transport of both proteins and organelles, it is not surprising that perturbation of microtubule-based transport may play a major role in the pathogenesis of AD.
MATERIALS AND METHODS:
We used the cDNA subtraction methodology and in vitro neural cell culture analyses to study the meaning of the brain site-specific gene expression pattern in cerebral tissue obtained from AD patients and also from control subjects at autopsy.
RESULTS:
We observed that cytoskeleton-associated proteins were down-regulated in AD subjects. We also noted an altered expression of the microtubule-associated protein 1B (MAP1B), the heat-shock protein (HSP)-90 (a key chaperone molecule), the tripartite motif-containing proteins (TRIM)-32/37 (an anti apoptotic enzyme with ubiquitin-protein ligase activity) and the Reticulon-3 (a modulator of the amyloid-precursor-protein (APP) cleavage) in AD brains. Additional molecular- and cell-biological studies revealed that small interfering RNA (siRNA)-mediated down-regulation of MAP1B expression leads to neuronal cell death in vitro.
CONCLUSION:
Altered expression of MAP1B, HSP90, TRIM32/37 and Reticulon-3 provides new clues by which the ubiquitin-proteasome-, the protein-chaperon- and the APP-processing systems are disturbed in AD, thus, leading to neuritic amyloid plaques and neurofibrillary tangles.

Alzheimer's disease (AD) is an aging-related neurodegenerative disorder characterized by irrevers... more Alzheimer's disease (AD) is an aging-related neurodegenerative disorder characterized by irreversible loss of higher cognitive functions. The disease is characterized by the presence of amyloid plaques and neurofibrillary tangles (NFT). In the current study we isolated from an intra-cerebral brain-site-specific (AD temporal lobe vs. AD occipital lobe) polymerase chain reaction (PCR)-select cDNA suppression subtractive hybridization (PCR-cDNA-SSH) expression analysis the novel gene P9TLDR, potentially a microtubule-associated protein involved in neuronal migration, with an altered expression pattern: down-regulated in the temporal lobe cortex of early stage AD brains. In an in vitro AD-related cell model, amyloid-β peptide (Aβ)-treated neurons, reduced P9TLDR expression correlated with increased tau protein phosphorylation. In conclusion, interference with the P9TLDR signalling pathways might be a therapeutic strategy for the treatment of AD.

he protein family of the neurotrophins (NTs) comprises structurally and functionally related mole... more he protein family of the neurotrophins (NTs) comprises structurally and functionally related molecules such as nerve growth factor (NGF) which influences the proliferation, differentiation, survival and death of neuronal cells. In addition to their established functions for cell survival, NTs also mediate higher brain activities such as learning and memory. Changes in NT expression levels have thus been implicated in neurological diseases such as Alzheimer's disease (AD), an age-related neurodegenerative disorder that is characterized by progressive loss of memory and deterioration of higher cognitive functions. The present review provides an overview of the functional role of NGF in neural stem cells and AD while pointing to a potential application of this peptide for the treatment of AD.

The number of dementia patients has been growing in recent years and dementia represents a signif... more The number of dementia patients has been growing in recent years and dementia represents a significant threat to aging people all over the world. Recent research has shown that the number of people affected by Alzheimer's disease (AD) and dementia is growing at an epidemic pace. The rapidly increasing financial and personal costs will affect the world's economies, health care systems, and many families. Researchers are now exploring a possible connection among AD, vascular dementia (VD), diabetes mellitus (type 2, T2DM) and cardiovascular diseases (CD). This correlation may be due to a strong association of cardiovascular risk factors with AD and VD, suggesting that these diseases share some biologic pathways. Since heart failure is associated with an increased risk of AD and VD, keeping the heart healthy may prove to keep the brain healthy as well. The risk for dementia is especially high when diabetes mellitus is comorbid with severe systolic hypertension or heart disease. In addition, the degree of coronary artery disease (CAD) is independently associated with cardinal neuropathological lesions of AD. Thus, the contribution of T2DM and CD to AD and VD implies that cardiovascular therapies may prove useful in preventing AD and dementia.

Alzheimer's disease (AD) is an age-related neurodegenerative disorder that is characterized by a ... more Alzheimer's disease (AD) is an age-related neurodegenerative disorder that is characterized by a progressive loss in memory and deterioration of the higher cognitive functions. The brain of an individual with AD exhibits extracellular senile plaques of aggregated amyloid-beta-peptide (Abeta), intracellular neurofibrillary tangles (NFTs) that consist of hyperphosphorylated tau protein (P-tau) and a profound loss of basal forebrain cholinergic neurons that innervate the hippocampus and the neocortex. Recent data obtained via genomics, proteomics and molecular genetics, have gleaned new information with regard to the physiological and pathophysiological functions of the amyloid precursor protein (APP) and its cleavage product Abeta. This review glances over several aspects that may play a major role in the pathogenesis of AD providing an insight into APP's and Abeta's interplay with other cellular systems.

We demonstrated that nerve growth factor (NGF) levels were increased in hippocampus and cortical ... more We demonstrated that nerve growth factor (NGF) levels were increased in hippocampus and cortical areas, as well as in cerebrospinal fluid (CSF) of patients with Alzheimer's disease (AD). Such increases may, at least in part, be due to a decreased expression of the NGF high affinity receptor trkA. Measurement of CSF levels of NGF may add to the repertoire of potential biochemical diagnostic markers in living AD patients.

Background: Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (... more Background: Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), and neurotrophin 4/5 (NT-4/5) are members of the neurotrophin gene family that support the survival of specific neuronal populations, including those that are affected by neurodegeneration in Alzheimer disease (AD).
OBJECTIVE:
To determine whether neurotrophin protein levels are altered in the AD-affected brain compared with control brains.
METHODS:
We quantitated protein levels of NGF, BDNF, NT-3, and NT-4/5, and calculated neurotrophin/NT-3 ratios in AD-affected postmortem hippocampus, frontal and parietal cortex, and cerebellum, and compared them with age-matched control tissue (patients with AD/controls: hippocampus, 9/9 cases; frontal cortex, 19/9; parietal cortex, 8/5; and cerebellum, 5/7, respectively). We applied highly sensitive and specific enzyme-linked immunosorbent assays in rapid-autopsy-derived brain tissue (mean+/-SD postmortem interval, 2. 57+/-1.75 h, n=71) to minimize postmortem proteolytic activity.
RESULTS:
Levels of BDNF were significantly reduced in hippocampus and parietal cortex (P<.001, and P<.01) as well as BDNF/NT-3 ratios in frontal and parietal cortices (P<.05, and P<.01) in the group with AD compared with the control group. Levels of NGF and NGF/NT-3 ratio were significantly elevated in the group with AD compared with the control group in the hippocampus and frontal cortex (P<.001). Levels of NT-4/5 and the NT-4/NT-3 ratio were slightly reduced in hippocampus and cerebellum in the group with AD compared with the control group (P<.05). In contrast, the levels of NT-3 were unchanged in all brain regions investigated.
CONCLUSION:
Decreased levels of BDNF may constitute a lack of trophic support and, thus, may contribute to the degeneration of specific neuronal populations in the AD-affected brain, including the basal forebrain cholinergic system.

Neurotrophin 3 (NT-3) is a member of the neurotrophin gene family which supports the survival of ... more Neurotrophin 3 (NT-3) is a member of the neurotrophin gene family which supports the survival of specific neurons. NT-3 was shown to prevent the death of adult central noradrenergic neurons in vivo, a neuronal population which is associated with the pathophysiology of major depression. We quantitated CSF levels of NT-3 in elderly patients with major depression (DE) and compared them to patients with Alzheimer's disease (AD), and mentally healthy control subjects (CTR). CSF levels of NT-3 were markedly and significantly elevated in the DE group, as compared to either the AD or the CTR group (P < 0.01, and P < 0.001, respectively). In terms of diagnostic accuracy, measurement of NT-3 levels in DE resulted in 73.9% sensitivity, and 89.7% specificity. Increased CSF levels of NT-3 may indicate a disturbance of the central noradrenergic system in patients with DE. NT-3 may constitute a biochemical candidate marker for clinical diagnosis and for the evaluation of therapeutic strategies in DE.

The authors quantitated CSF levels of nerve growth factor (NGF) in patients with AD, nondemented ... more The authors quantitated CSF levels of nerve growth factor (NGF) in patients with AD, nondemented control subjects (CTR), and age-matched patients with major depression (DE). CSF levels of NGF were markedly higher in the AD group than in both the CTR and DE groups (p < 0.01 and p < 0.001). Increased CSF levels of NGF in AD patients may reflect reported accumulation of NGF in the AD brain and may constitute a candidate marker for clinical diagnosis and therapeutic monitoring.

The cholinergic neurons of the basal forebrain system are sensitive to nerve growth factor (NGF),... more The cholinergic neurons of the basal forebrain system are sensitive to nerve growth factor (NGF), a member of the neurotrophin gene family. Since the cholinergic system is affected early in the course of Alzheimer's disease (AD), it was hypothesized that a deficit in NGF, e.g. reduced neurotrophin uptake by specific receptors, may play a role in neuronal cell death in AD. We quantitated mRNA levels of neurotrophins (NGF, BDNF, NT-3, NT-4/5) and their receptors (trkA, trkB, trkC, p75) in AD postmortem parietal cortex (n = 16) and cerebellum (n = 11). We applied highly sensitive reverse transcription-polymerase chain reaction (RT-PCR) in rapid autopsy derived brain tissue (mean postmortem delay 147+/-96 min., n = 53) to minimize postmortem mRNA variations. In the AD parietal cortex trkA mRNA levels were more than two times lower as compared to controls (n = 16, mean+/-SEM 0.26+/-0.07 units/S12, range, 0-1.78, and n = 11, 0.59+/-0.10 units/S12, range, 0.17-1.10, respectively, P = 0.015). TrkA mRNA levels did not appear to be altered in the AD cerebellum as compared to normal human cerebellum. NGF, BDNF, NT-3, NT-4/5, as well as trkB, trkC and p75 mRNA levels were unchanged in AD parietal cortex and cerebellum as compared to controls. This finding suggests that a reduced expression of the trkA receptor may contribute to impaired NGF-trkA signalling and a reduced transport of NGF in cholinergic neurons. These results reveal a central specific role of the high affinity NGF receptor during neurodegeneration in AD.

Inflammatory processes involving reactive microglia, e.g., those associated with beta-amyloid con... more Inflammatory processes involving reactive microglia, e.g., those associated with beta-amyloid containing neuritic and core plaques in Alzheimer's disease, appear to contribute to neuronal degeneration in the CNS. The fact that increased nerve growth factor (NGF) protein levels were found throughout brains of Alzheimer's disease patients led us to investigate neurotrophin synthesis in a human microglial cell line showing typical properties of human microglial cells, including expression of neurotrophins such as NGF, as well as the NGF receptor trkA and the low-affinity neurotrophin receptor p75. We found that the cytokines interleukin-1beta and tumor necrosis factor-alpha synergistically stimulate microglial NGF transcription and protein release. Moreover, exposure of microglial cells to complement factor C3a induces NGF expression. To assess the role of the transcription factor nuclear factor-kappaB (NF-kappaB) in inflammatory mediator-induced microglial NGF expression, the effect of the NF-kappaB inhibitor pyrrolidine dithiocarbamate (PDTC) was analyzed. In the presence of PDTC, a dose-dependent inhibition of cytokine-activated NGF expression occurred. In contrast, the C3a-dependent stimulation of NGF synthesis was not influenced by PDTC. In addition, microglial neurotoxicity-mediating beta-amyloid peptides A beta(1-40) and A beta(1-42) failed to alter NGF synthesis, whereas A beta(25-35) specifically induced NF-kappaB-dependent microglial NGF expression. In conclusion, inflammatory signals (cytokines and complement factors), as well as A beta(25-35), are potent stimulators of human microglial NGF synthesis involving NF-kappaB-dependent and -independent mechanisms. Microglial secretion of neurotrophins appears to be involved in early processes of neuronal regeneration.

The β-amyloid precursor protein (APP) is a type I transmembrane protein whose functions and metab... more The β-amyloid precursor protein (APP) is a type I transmembrane protein whose functions and metabolic processing have been implicated in the pathogenesis of Alzheimer's disease (AD). APP's physical resemblance as a glycosylated receptor and the presence of several conserved motifs which are characteristics of a membrane-associated receptor has prompted interest to study and understand the role of downstream signaling events mediated by the activation of APP during both physiological and pathological conditions. Efforts to elucidate the mechanisms of APP signaling have not been conclusive. In order to further characterize the intracellular signaling activities of APP, we have constructed a chimeric APP receptor which is made up of the extracellular domain of human tumor necrosis factor receptor superfamily, member 1B (TNFRSF1B), the membrane-spanning segment of human APP(770) isoform's β-cleaved carboxyl-terminal fragment (C99), and the green fluorescent protein (GFP) tagged to the carboxyl-terminus of C99. The mammalian rat pheochromocytoma (PC12) cell line is used as the cellular model for transfection of this TNFRSF1B-APP-GFP (TAF) fusion receptor. Ligand-specific stimulation of this TAF receptor using human recombinant TNF reveals the induction of cytoplasmic TAF phosphorylation on Thr743 (numbering for APP(770) isoform) and an elevated release of APP intracellular domains (AICDs). Time-lapse microscopy shows the trafficking of GFP-tagged AICDs into the nucleus upon TAF receptor activation. The increased presence of AICDs is believed to suppress neuronal differentiation of PC12 cells in response to nerve growth factor treatment, by negatively regulating the levels of p53, cyclin D1 and phosphorylation of the signal transducer and activator of transcription STAT3.

Ischemic stroke, still lacking an effective neuroprotective therapy is the third leading cause of... more Ischemic stroke, still lacking an effective neuroprotective therapy is the third leading cause of global mortality and morbidity. Here, we have applied an 8-plex iTRAQ-based 2D-LC-MS/MS strategy to study the commonly regulated infarct proteome from three different brain regions (putamen, thalamus and the parietal lobe) of female Japanese patients. Infarcts were compared with age-, post-mortem interval- and location-matched control specimens. The iTRAQ experiment confidently identified 1520 proteins with 0.1% false discovery rate. Bioinformatics data mining and immunochemical validation of pivotal perturbed proteins revealed a global failure of the cellular energy metabolism in the infarcted tissues as seen by the parallel down-regulation of proteins related to glycolysis, pyruvate dehydrogenase complex, TCA cycle and oxidative phosphorylation. The concomitant down-regulation of all participating proteins (SLC25A11, SLC25A12, GOT2 and MDH2) of malate-aspartate shuttle might be responsible for the metabolic in-coordination between the cytosol and mitochondria resulting in the failure of energy metabolism. The levels of proteins related to reactive gliosis (VIM, GFAP) and anti-inflammatory response (ANXA1, ANXA2) showed an increasing trend. The elevation of ferritin (FTL, FTH1) may indicate an iron-mediated oxidative imbalance aggravating the mitochondrial failure and neurotoxicity. The deregulated proteins could be useful as potential therapeutic targets or biomarkers for ischemic stroke.
BIOLOGICAL SIGNIFICANCE:
Clinical proteomics of stroke has been lagging behind other areas of clinical proteomics like Alzheimer's disease or schizophrenia. Our study is the first quantitative clinical proteomics study where iTRAQ-2D-LC-MS/MS has been utilized in the area of ischemic stroke to obtain a comparative profile of human ischemic infarcts and age-, sex-, location- and post-mortem interval-matched control brain specimens. Different pathological attributes of ischemic stroke well-known through basic and pre-clinical research such as failure of cellular energy metabolism, reactive gliosis, activation of anti-inflammatory response and aberrant iron metabolism have been observed at the bedside. Our dataset could act as a reference for similar studies done in the future using ischemic brain samples from various brain banks across the world. A meta-analysis of these studies could help to map the pathological proteome specific to ischemic stroke that will guide the scientific community to better evaluate the pros and cons of the pre-clinical models for efficacy and mechanistic studies. Infarct being the core of injury should have the most intense regulation for several key proteins involved in the pathophysiology of ischemic stroke. Hence, a part of the up-regulated proteome could leak into the general circulation that may offer candidates of interest as potential biomarkers. In support of our proposed hypothesis, we report ferritin in the current study as one of the most elevated proteins in the infarct, which has been documented as a biomarker in the context of ischemic stroke by an independent study. Overall, our approach has the potential to identify probable therapeutic targets and biomarkers in the area of ischemic stroke.

Cerebral ischemia or stroke, an acute neurological injury lacking an effective therapy, is the se... more Cerebral ischemia or stroke, an acute neurological injury lacking an effective therapy, is the second leading cause of death globally. The unmet need in stroke research is to identify viable targets and to understand their interplay during the temporal evolution of ischemia/reperfusion (I/R) injury. Here we report a temporal signature of the ischemic hemisphere revealed by the isobaric tag for relative and absolute quantification (iTRAQ)-based 2D-LC-MS/MS strategy in an in vivo middle cerebral artery occlusion (MCAO) model of focal cerebral I/R injury. To recapitulate clinical stroke, two hours of MCAO was followed by 0, 4, and 24 h of reperfusion to capture ischemia with an acute and subacute durations of reperfusion injury. The subsequent iTRAQ experiment identified 2242 proteins from the ischemic hemisphere with <1.0% false discovery rate. Data mining revealed that (1) about 2.7% of detected proteins were temporally perturbed having an involvement in the energy metabolism (Pygb, Atp5b), glutamate excitotoxicity (Slc1a3, Glud1), neuro-inflammation (Tf, C3, Alb), and cerebral plasticity (Gfap, Vim, Gap43); (2) astrocytes participated actively in the neurometabolic coupling underlining the importance of a cerebro-protective rather than a neuro-protective approach; and (3) hyper-acute yet progressive opening of the blood brain barrier (BBB), accompanied by stimulation of an innate immune response and late activation of a regenerative response, which provides an extended therapeutic window for intervention. Several regulated proteins (Caskin1, Shank3, Kpnb1, Uchl1, Mtap6, Epb4.1l1, Apba1, and Ube1x) novel in the context of stroke were also discovered. In conclusion, our result supports a dynamic multitarget therapy rather than the traditional approach of a unilateral and sustained modulation of a single target to address the phasic regulation of an ischemic proteome.

Cerebral ischemia is a major cause of death and long-term disability worldwide. Ischemic penumbra... more Cerebral ischemia is a major cause of death and long-term disability worldwide. Ischemic penumbra, the electrically silent but metabolically viable perifocal brain tissue, is the target for the much elusive stroke therapy. To characterize the molecular events of the dynamic penumbra, we applied an iTRAQ-based shotgun proteomic approach in an in vitro neuronal model, using the rat B104 neuroblastoma cell line. Various functional and cytometric assays were performed to establish the relevant time-point and conditions for ischemia to recapitulate the pathology of the penumbra. Two replicate iTRAQ experiments identified 1796 and 1566 proteins, respectively (<or=1.0% false discovery rate). Mining of proteomic data indicated the up-regulation of proteins involved in ammoniagenesis, antiapoptotic, anti-inflammatory and mitochondrial heat shock response and down-regulation of proteins pertaining to antioxidative defense and protein metabolism. Additionally, many proteins (for instance, park7 and VAP-A) involved in the chronic neurological disorders (such as Alzheimer's disease, Parkinson's disease or Bipolar disorder) were also regulated in this model of acute neuronal injury. Our results also provide preliminary evidence about the presence of a relative glucose paradox under in vitro conditions indicating possible application of this cell system to study the mechanisms of transient protection induced by concomitant glucose deprivation under hypoxia. In conclusion, our study shows the potential application of iTRAQ-based quantitative proteomics for the elucidation of pathophysiology and the discovery of novel therapeutic targets in the field of neuroproteomics.

ACS Omega, 6 (29), 19045−19057, 2021

Bioactive constituents from natural sources are of great interest as alternatives to synthetic co... more Bioactive constituents from natural sources are of great interest as alternatives to synthetic compounds for the treatment of various diseases, including diabetes mellitus. In the present study, phytochemicals present in Leucaena leucocephala (Lam.) De Wit leaves were identified by gas chromatography−mass spectrometry and further examined by qualitative and quantitative methods. α-Amylase enzyme activity assays were performed and revealed that L. leucocephala (Lam.) De Wit leaf extract inhibited enzyme activity in a dose-dependent manner, with efficacy similar to that of the standard α-amylase inhibitor acarbose. To determine which phytochemicals were involved in α-amylase enzyme inhibition, in silico virtual screening of the absorption, distribution, metabolism, excretion, and toxicity properties was performed and pharmacophore dynamics were assessed. We identified hexadecenoic acid and oleic acid ((Z)-octadec-9-enoic acid) as α-amylase inhibitors. The binding stability of α-amylase to those two fatty acids was confirmed in silico by molecular docking and a molecular dynamics simulation performed for 100 ns. Together, our findings indicate that L. leucocephala (Lam.) De Wit-derived hexadecanoic acid and oleic acid are natural product-based antidiabetic compounds that can potentially be used to manage diabetes mellitus.

We evaluated the production and characterization of endoglucanase from Ganoderma lucidum using di... more We evaluated the production and characterization of endoglucanase from Ganoderma lucidum using different lignocellulose biomasses. We purified a novel carboxymethyl cellulose (CMC) hydrolyzing endoglucanase from the white-rot fungus G. lucidum when the medium was supplemented with 1% (w/v) wheat bran. Endoglucanase was purified 12.5-fold via ammonium sulfate fractionation, Sephadex G-100, and Q-Sepharose column chromatography with a final yield of 15%. SDS-PAGE analysis revealed that the endoglucanase had a molecular mass of 64.0 kDa. The optimal activity of purified endoglucanase was at pH 5.0 and 35 °C, though it was stable between pH 4.0–7.0 and temperatures of 30–60 °C. The purified enzyme was specific to CMC as a suitable substrate. The metal ions Hg2+, Fe2+, and Cr2+ inhibited enzyme activity, while Ca2+, Mg2+, and Mn2+ enhanced enzyme activity. The endoglucanase showed high activity and stability in the presence of different surfactants and non-polar hydrophobic organic solvents. This endoglucanase is tolerant to high temperature, metal ions, surfactants, and solvents, suggesting that it is appropriate for use in biomass conversion for biofuel production under harsh environmental conditions.

The development of alternative energy sources by applying lignocellulose-based biofuel technology... more The development of alternative energy sources by applying lignocellulose-based biofuel technology is critically important because of the depletion of fossil fuel resources, rising fossil fuel prices, security issues regarding the fossil fuel supply, and environmental issues. White-rot fungi have received much attention in recent years for their valuable enzyme systems that effectively degrade lignocellulosic biomasses. These fungi have powerful extracellular oxidative and hydrolytic enzymes that degrade lignin and cellulose biopolymers, respectively. Lignocellulosic biomasses from either agricultural or forestry wastes are abundant, low-cost feedstock alternatives in nature but require hydrolysis into simple sugars for biofuel production. This review provides a complete overview of the different lignocellulose biomasses and their chemical compositions. In addition, a complete list of the white-rot fungi-derived lignocellulolytic enzymes that have been identified and their molecular structures, mechanism of action in lignocellulose hydrolysis, and biochemical properties is summarized in detail. These enzymes include ligninolytic enzymes (laccase, manganese peroxidase, lignin peroxidase, and versatile peroxidase) and cellulolytic enzymes (endo-glucanase, cellobiohydrolase, and beta-glucosidase). The use of these fungi for low-cost lignocellulolytic enzyme production might be attractive for biofuel production.

Plasmodium falciparum (Pf)-mediated malaria is one of the most devastating diseases in the world,... more Plasmodium falciparum (Pf)-mediated malaria is one of the most devastating diseases in the world, and the search for suitable antimalarial drugs remains an extraordinary challenge for scientists working in this area. Novel unconventional approaches could reveal new potential targets that may be useful for the treatment of malaria. We used a bioinformatics approach to analyze the entire genome of the Pf3D7 strain. Because the carbon (C-) content is a pivotal parameter that determines the hydrophobicity of a protein, which in turn controls protein folding and function, we analyzed the entire Pf3D7 proteome based on the gene's thymine (T)-controlled amino acid expression. Our data disclose a total of 14 proteins encoded by chromosome-4 and chromosome-9 that have an outstanding T-encoded and C-controlled hydrophobic character. The identification of these proteins could open new pivotal drug-targeting avenues.

Although GlaxoSmithKline is on the way to launch the new vaccine candidate 'RTS, S', the search f... more Although GlaxoSmithKline is on the way to launch the new vaccine candidate 'RTS, S', the search for suitable antimalarial drugs still remains an exceeding challenge because Plasmodium falciparum-mediated malaria is one of the most lethal diseases in the world. Novel innovative ideas are required to identify new potential molecular targets to be able to fight this lethal parasite efficiently. We used an unconventional bioinformatics approach to analyze the entire genome and proteome of the Pf3D7 strain. Because the oxygen (O-) content is a decisive parameter that determines the function of a protein, we analyzed the entire Pf3D7 proteome based on O-containing amino acid expression. Our data disclose a total of four proteins encoded by chromosome (Chr)-4 and Chr-9 that have an outstanding O-controlled character. The identification of the biological significance of these proteins could eventually lead to new vital drug targets.

We show for the first-time Ganoderma lucidum laccase enzyme production using medium containing 3%... more We show for the first-time Ganoderma lucidum laccase enzyme production using medium containing 3%
(v/v) ethanol, which enhanced the enzyme production up to 14.1 folds. A more than 400-folds increase
could be achieved if grown in the presence of the novel lignocellulosic biomass tamarind shell plus
ethanol (3%, v/v), CuSO4 (0.4 mM) and gallic acid (1 mM). A 38.3 kDa laccase enzyme was purified from
the initial protein preparation with an overall yield of 32% using Sephadex G-100 and DEAE-cellulose column
chromatography. The enzyme was identified through MALDI-TOF/TOF tandem mass spectrometry
(MS/MS) as G. lucidum laccase-3. This enzyme exerted its optimal activity at a pH of 5 and a temperature
of 55 ◦C with ABTS (2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) as an ideal substrate. The
catalytic efficiencies (kcat/Km) determined for ABTS and guaiacol were 11.5 × 105 and 3.9 × 105 s−1M−1, respectively. The G. lucidum laccase decolorized various textile dyes and industrial textile dye effluent up to 90% and 97%, respectively.

Harmful environmental issues of fossil-fuels and concerns about petroleum supplies have spurred t... more Harmful environmental issues of fossil-fuels and concerns about petroleum supplies have spurred the search for renewable alternative fuels such as biofuel. Agricultural crop residues represent an abundant renewable resource for future biofuel. To be a viable alternative, a biofuel should provide a net energy gain, have environmental benefits, be economically feasible, and should also be producible in large quantities without reducing food supplies. We used these criteria to evaluate the white rot basidiomycota-derived fungus Ganoderma lucidum that secretes substantial amounts of hydrolytic and oxidative enzymes useful for the degradation of lignocellulosic biomass that were not described hitherto. The current bottleneck of lignocellulosic biofuel production is the hydrolysis of biomass to sugar. To understand the enzymatic hydrolysis of complex biomasses, we cultured G. lucidum with sugarcane bagasse as substrate and qualitatively analyzed the entire secretome. The secreted lignocellulolytic enzymes were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and diverse enzymes were found, of which several were novel lignocellulosic biomass hydrolyzing enzymes. We further explored G. lucidum-derived cellulose, hemicellulose and lignin degrading enzymes as valuable enzymes for the second generation of biofuel obtained from a lignocellulose substrate such as sugarcane bagasse.

FAM72 is a novel neuronal progenitor cell (NPC) self-renewal supporting protein expressed under p... more FAM72 is a novel neuronal progenitor cell (NPC) self-renewal supporting protein expressed under physiological conditions at low levels in other tissues. Accumulating data indicate the potential pivotal tumourigenic effects of FAM72. Our in silico human genome-wide analysis (GWA) revealed that the FAM72 gene family consists of four human-specific paralogous members, all of which are located on chromosome (chr) 1. Unique asymmetric FAM72 segmental gene duplications are most likely to have occurred in conjunction with the paired genomic neighbour SRGAP2 (SLIT-ROBO Rho GTPase activating protein), as both genes have four paralogues in humans but only one vertebra-emerging orthologue in all other species. No species with two or three FAM72/SRGAP2 gene pairs could be identified, and the four exclusively human-defining ohnologues, with different mutation patterns in Homo neanderthalensis and Denisova hominin, may remain under epigenetic control through long non-coding (lnc) RNAs.

FAM72A (p17) is a novel neuronal protein that has been linked to tumorigenic effects in non-neuro... more FAM72A (p17) is a novel neuronal protein that has been linked to tumorigenic effects in non-neuronal tissue. Using state of the art in silico physicochemical analyses (e.g., I-TASSER, RaptorX, and Modeller), we determined the three-dimensional (3D) protein structure of FAM72A and further identified potential ligand-protein interactions. Our data indicate a Zn(2+)/Fe(3+)-containing 3D protein structure, based on a 3GA3_A model template, which potentially interacts with the organic molecule RSM ((2s)-2-(acetylamino)-N-methyl-4-[(R)-methylsulfinyl] butanamide). The discovery of RSM may serve as potential lead for further anti-FAM72A drug screening tests in the pharmaceutical industry because interference with FAM72A's activities via RSM-related molecules might be a novel option to influence the tumor suppressor protein p53 signaling pathways for the treatment of various types of cancers.

Thymine is the one and only base transcribed into uracil during production of proteins. Thymine i... more Thymine is the one and only base transcribed into uracil during production of proteins. Thymine in DNA and uracil in mRNA plays a major role in producing proteins with appropriate carbon content for stability and activity. Thymine distribution is different frames of coding nucleic acids are investigated statistically. The results confirm that frame 1 supposed to have definite thymine content. Frame 3 prefers to have least thymine content. Frames 4 & 5 maintain some degree of thymine while 2 & 6 have a variable fraction of thymine.

Introduction Glioblastoma multiform (GBM) is a neural stem cell (NSC)-derived malignant brain tum... more Introduction Glioblastoma multiform (GBM) is a neural stem cell (NSC)-derived malignant brain tumor with complex
genetic alterations challenging clinical treatments. FAM72 is a NSC-specific protein comprised of four paralogous genes
(FAM72 A-D) in the human genome, but its functional tumorigenic significance is unclear.
Methods We conducted an in-depth expression and somatic mutation data analysis of FAM72 (A-D) in GBM using the
comprehensive human clinical cancer study database cBioPortal [including The Cancer Genome Atlas (TCGA)].
Results We established a FAM72 transcription profile across TCGA correlated with the expression of the proliferative
marker MKI67 and a tissue-specific gene-mutation signature represented by pivotal genes involved in driving the cell cycle.
FAM72 paralogs are overexpressed in cancer cells, specifically correlating with the mitotic cell cycle genes ASPM, KIF14,
KIF23, CENPE, CENPE, CEP55, SGO1, and BUB1, thereby contributing to centrosome and mitotic spindle formation.
FAM72 expression correlation identifies a novel GBM-specific gene set (SCN9A, MXRA5, ADAM29, KDR, LRP1B, and
PIK3C2G) in the de novo pathway of primary GBM predestined as viable targets for therapeutics.
Conclusion Our newly identified primary GBM-specific gene-mutation signature, along with FAM72, could thus provide a
new basis for prognostic biomarkers for diagnostics of GBM and could serve as potential therapeutic targets.

Adrenocortical carcinoma (ACC) is a rare and aggressive tumor whose molecular signaling pathways ... more Adrenocortical carcinoma (ACC) is a rare and aggressive tumor whose molecular signaling pathways are not fully understood. Using an in-silico clinical data analysis approach we retrieved human gene mutation data from the highly reputed Cancer Genome Atlas (TCGA). ACC-specific gene mutations were correlated with proliferation marker FAM72 expression and Mutsig along with the algorithmic implementation of the 20/20 rule were used to validate their oncogenic potential. The newly identified oncogenic driver gene set (ZFPM1, LRIG1, CRIPAK, ZNF517, GARS and DGKZ), specifically and most repeatedly mutated in ACC, is involved in tumor suppression and cellular proliferation and thus could be useful for the prognosis and development of therapeutic approaches for the treatment of ACC.

RAS protein is a small G protein linked to multiple G protein-coupled receptor (GPCR) signaling c... more RAS protein is a small G protein linked to multiple G protein-coupled receptor (GPCR) signaling cascades and is responsible for various types of cancer, but to this day, Ras is considered " undruggable. " Multiple alternative regulators of G protein signaling (RGS) pathways have become the focus of ongoing efforts to identify new cancer therapeutics. We analyzed human cancer genome datasets and describe p60TRP, a recently identified GPCR-associated sorting protein (GPRASP), and its role in various types of cancer. We found that some regions of p60TRP were more prone to specific mutations, with two hotspots for mutations at E15 and E171.

Keywords: FAM72 neuron cancer homo p17 FAM72 is a novel neuronal progenitor cell (NPC) self-renew... more Keywords: FAM72 neuron cancer homo p17 FAM72 is a novel neuronal progenitor cell (NPC) self-renewal supporting protein expressed under physiological conditions at low levels in other tissues. Accumulating data indicate the potential pivotal tumourigenic effects of FAM72. Our in silico human genome-wide analysis (GWA) revealed that the FAM72 gene family consists of four human-specific paralogous members, all of which are located on chromosome (chr) 1. Unique asymmetric FAM72 segmental gene duplications are most likely to have occurred in conjunction with the paired genomic neighbour SRGAP2 (SLIT-ROBO Rho GTPase activating protein), as both genes have four paralogues in humans but only one vertebra-emerging orthologue in all other species. No species with two or three FAM72/SRGAP2 gene pairs could be identified, and the four exclusively human-defining ohnologues, with different mutation patterns in Homo neanderthalensis and Denisova hominin, may remain under epigenetic control through long non-coding (lnc) RNAs.

Active cell death ('apoptosis' or 'programmed cell death') is essential in the development and ho... more Active cell death ('apoptosis' or 'programmed cell death') is essential in the development and homeostasis of multicellular organisms and abnormal inhibition of apoptosis is an indicator of cancer and autoimmune diseases, whereas excessive cell death is implicated in neurodegenerative disorders such as Alzheimer's disease (AD). Here we demonstrate new isoforms of the rat homologue of the drosophila tumor suppressor l(2)tid gene (rTid-1). Moreover, we show that rTid-1 interacts isoform-specifically with the heat-shock-cognate-glucose-regulated protein hscGRP75 and neither induces nor inhibits directly neuronal apoptosis. This finding points to a pivotal role of Tid-1 in the control of cellular survival.

P17 is a novel neuronal protein expressed under physiological conditions only at very low levels ... more P17 is a novel neuronal protein expressed under physiological conditions only at very low levels in other tissues. Accumulating data indicate its crucial involvement in tumorigenic effects. Using molecular, cellular, and biocomputational methods, the current study unraveled p17 mode of action. Data indicate that mitochondria-associated p17 interacts with the proteins TMEM115, YPEL3, ERP44, CDK5RAP, and NNAT. Moreover, p17 drives the cell cycle into the G0/G1 phase and enhances survival of proliferating cells. Interference with p17 activities thus might become a novel option to influence also the tumor suppressor protein p53 signaling pathways for the treatment of tumors.

Although the regulation of several Bcl-2 family molecules, including Puma, Noxa, Bax, and Bid, by... more Although the regulation of several Bcl-2 family molecules, including Puma, Noxa, Bax, and Bid, by p53 has been studied intensively, the interplay between Bad (Bcl-2 antagonist of cell death) and p53 has not yet been reported thus far. Here, we report that p53 activates Bad transcription and expression through binding to a short conserved sequence located approximately 6.6 kb upstream of the translation start point. We also demonstrate that Bad physically interacts with cytoplasmic p53, thereby preventing p53 from entering the nucleus and resulting in reduced transcription of Bad. Moreover, Bad is able to direct p53 to the mitochondria and forms a p53/Bad complex at the mitochondria. Two lines of evidences support this hypothesis: first, when mitochondria purified from p53-deficient H1299 cells are incubated with p53 and either wild-type (wt) Bad or mutant Bad (this mutant binds p53 yet is unable to migrate to mitochondria), p53 can be detected only in mitochondria incubated with wt Bad and not in those incubated with mutant Bad; second, knockdown of Bad expression reduces mitochondrial localization of p53. The mitochondrial p53/Bad complex promotes apoptosis via activation and oligomerization of Bak. Elimination of Bad expression by RNA interference notably attenuates apoptosis induced by etoposide. Hence, our collective data provide the first evidence that Bad plays dual roles in both p53 transcription-dependent and -independent pathways.

Activation of the apical caspase-8 is crucial to the extrinsic apoptotic pathway. Although the de... more Activation of the apical caspase-8 is crucial to the extrinsic apoptotic pathway. Although the death effector domain (DED) of caspase-8 has been reported to be involved in death-inducing signaling complex formation, the detailed mechanism of how DED functions in regulating apoptosis remains largely unknown. Here, we demonstrate that the prodomain of the caspase-8/Mch5 can be further cleaved between two tandemly repeated DEDs (DEDa-DEDb) at the amino-acid residue Asp129 by caspase-8 itself. The DEDa fragment generated from the endogenous caspase-8 was detected in isolated nucleoli upon treatment with TRAIL (tumor necrosis factor-related apoptosis-inducing ligand). Cleaved DEDa appears to translocate into the nucleus by association with extracellular signal-regulated protein kinases-1/2 (ERK1/2). Elimination of ERK1/2 expression by RNA interference resulted in a significant attenuation of nuclear entry of DEDa and reduced caspase-8-dependent apoptosis. In the nucleus, DEDa interacts with TOPORS, a p53 and topoisomerase I binding protein, and possibly displaces p53 from TOPORS, allowing p53 to stimulate caspase-8 gene expression. In summary, we postulate a positive feedback loop involving DEDa, which enables the continual replenishment of procaspase-8 during apoptosis.

Livin, a member of the inhibitor of apoptosis protein (IAP) family, encodes a protein containing ... more Livin, a member of the inhibitor of apoptosis protein (IAP) family, encodes a protein containing a single baculoviral IAP repeat (BIR) domain and a COOH-terminal RING finger domain. It has been reported that Livin directly interacts with caspase-3 and -7 in vitro and caspase-9 in vivo via its BIR domain and is negatively regulated by Smac/DIABLO. Nonetheless, the detailed mechanism underlying its antiapoptotic function has not yet been fully characterized. In this report, we provide, for the first time, the evidence that Livin can act as an E3 ubiquitin ligase for targeting the degradation of Smac/DIABLO. Both BIR domain and RING finger domain of Livin are required for this degradation in vitro and in vivo. We also demonstrate that Livin is an unstable protein with a half-life of less than 4 h in living cells. The RING domain of Livin promotes its auto-ubiquitination, whereas the BIR domain is likely to display degradation-inhibitory activity. Mutation in the Livin BIR domain greatly enhances its instability and nullifies its binding to Smac/DIABLO, resulting in a reduced antiapoptosis inhibition. Our findings provide a novel function of Livin: it exhibits E3 ubiquitin ligase activity to degrade the pivotal apoptotic regulator Smac/DIABLO through the ubiquitin-proteasome pathway.

Mani (myelin-associated neurite-outgrowth inhibitor) protein is implicated in both axonal guidanc... more Mani (myelin-associated neurite-outgrowth inhibitor) protein is implicated in both axonal guidance and axonal regeneration after central nervous system (CNS) injury. Here, we applied a neurite outgrowth assay, coupled with a siRNA-driven investigation and immunocytochemistry, to unveil Mani's axonal outgrowth inhibitory effect in embryonic rat cortical primary neurons in vitro. We further demonstrate Mani's neuronal localization in comparison with a principal subunit, Cdc27, of the anaphase promoting complex (APC). Considering the protein structure of Mani obtained via a series of bio-computational studies, we propose a Cdc27-Mani-APC-related signalling pathway may be involved in CNS axon regeneration.

Neuronal regeneration and axonal re-growth in the injured mammalian central nervous system remain... more Neuronal regeneration and axonal re-growth in the injured mammalian central nervous system remains an unsolved field. To date, three myelin-associated proteins [Nogo or reticulon 4 (RTN4), myelin-associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMG)] are known to inhibit axonal regeneration via activation of the neuronal glycosylphosphatidylinositol-anchored Nogo receptor [NgR, together with p75 neurotrophin receptor (p75NTR) and Lingo-1]. In the present study we describe the novel protein MANI (myelin-associated neurite-outgrowth inhibitor) that localizes to neural membranes. Functional characterization of MANI overexpressing neural stem cells (NSCs) revealed that the protein promotes differentiation into catecholaminergic neurons. Yeast two-hybrid screening and co-immunoprecipitation experiments confirmed the cell division cycle protein 27 (Cdc27) as an interacting partner of Mani. The analyses of Mani-overexpressing PC12 cells demonstrated that Mani retards neuronal axonal growth as a positive effector of Cdc27 expression and activity. We show that knockdown of Cdc27, a component of the anaphase-promoting complex (APC), leads to enhanced neurite outgrowth. Our finding describes the novel MANI-Cdc27-APC pathway as an important cascade that prevents neurons from extending axons, thus providing implications for the potential treatment of neurodegenerative diseases.

In this study we show that single, physiologically-active and non-convulsive doses of the three G... more In this study we show that single, physiologically-active and non-convulsive doses of the three GABA(B) receptor antagonists CGP 36742, CGP 56433A and CGP 56999A increase NGF and BDNF mRNA levels by 200-400% and protein levels by 200-250% in rat neocortex, hippocampus as well as spinal cord. In all areas examined the increase in NGF protein preceded that of BDNF. Peak levels of both neurotrophins are transient and occur between 24 and 72 h, depending on the region. In contrast, NT-3 protein concentrations in the neocortex and hippocampus were decreased significantly to 50% of control values within 48-96 h. The decrease in the spinal cord was less than 30% and did not reach significant levels. These data clearly demonstrate that GABA(B) receptor antagonists induce a specific neurotrophin expression in the central nervous system at physiologically relevant doses, as opposed to the extreme conditions of seizure paradigms. The results are in line with the concept that neuronal neurotrophin synthesis and release in brain are controlled by afferent nerve activity. GABA(B) receptor antagonists could therefore be a valuable new approach to selectively increase endogenous neurotrophin levels in the central nervous system.

The novel protein p33MONOX (p33Monooxygenase) was over-expressed in neuroblastoma cells demonstra... more The novel protein p33MONOX (p33Monooxygenase) was over-expressed in neuroblastoma cells demonstrating its inhibitory effect on the phosphorylation of the App (amyloid precursor protein) and Bcl2 (B-cell lymphoma 2) proteins but mediating higher activation of Mapk1/3 (mitogen-activated protein kinase 1/3). We employed a variety of cell biology techniques to show the localization of p33MONOX to the cytoplasm of pyramidal neurons in the mouse brain hippocampus. We also carried out a yeast-two-hybrid screening plus co-immunoprecipitation and bio-informatics to determine COBRA1 (cofactor of BRCA1 (breast cancer type 1)), NOL12 (nucleolar protein 12), and PRNP (prion protein) as p33MONOX-interacting proteins. Bio-computational analyses revealed a flavine-containing monooxygenase (FMO)-1 motif, thus linking p33MONOX to a group of previously characterized proteins, the MICALs (molecule interacting with CasL). Concluding, p33MONOX might regulate pre- and post-transcriptional control of dynamic processes related to growth cone guidance.

Apoptosis is an active form of cell death that is carried out by proteins that are designed to ki... more Apoptosis is an active form of cell death that is carried out by proteins that are designed to kill the cell during normal mammalian development and tissue homeostasis. Cell death by apoptosis comprises a sequence of events leading to the activation of caspases which execute the fragmentation of the cellular protein and DNA leading to disintegration of the cell. This physiological neuronal apoptosis allows the nervous system to eliminate excess neurons. In addition, apoptotic cell death occurs in a variety of neuronal degeneration such as Alzheimer's disease. Here we describe second mitochondria-derived activator of caspases/Diablo as a new interacting protein of CGI-94 (comparative gene identification-94) which itself is probably involved in degenerative processes of Alzheimer's disease. Our findings that CGI-94 interacts with second mitochondria-derived activator of caspases/Diablo, inhibits nerve growth factor-induced neurite outgrowth and that its neuronal expression leads to cell death point to its pivotal role in the control of cellular survival. In conclusion, CGI-94 appears to be involved in processes of neuronal degeneration.

The treatment of Alzheimer's disease (AD) remains a major challenge because of the incomplete und... more The treatment of Alzheimer's disease (AD) remains a major challenge because of the incomplete understanding of the triggering events that lead to the selective neurodegeneration characteristic of AD brains. Here we describe a new protein, CGI-94, that is down-regulated at the mRNA level in the hippocampus of early stage AD brain. Transfection experiments with CGI-94 as a green fluorescent protein (GFP)-fusion-protein show that this protein is translocated into the nucleus of the cell. The finding that this protein, which has a bipartite nuclear localization signal, is also observed in the cytoplasm and extracellular space points to a multifunctional protein. Immunohistochemical analyses reveal that CGI-94 is mainly expressed in neurons of the hippocampal formation and the cortex but not in the cerebellar nucleus. In conclusion, the expression of the nucleolar phosphoprotein CGI-94 appears to be disturbed in early processes of neuronal degeneration.

Active cell death (‘apoptosis’ or ‘programmed cell death’) is essential in the development and ho... more Active cell death (‘apoptosis’ or ‘programmed cell death’) is essential in the development and homeostasis of multicellular organisms and abnormal inhibition of apoptosis is an indicator of cancer and autoimmune diseases, whereas excessive cell death is implicated in neurodegenerative disorders such as Alzheimer’s disease (AD). Here we show that Smac/Diablo (second mitochondria-derived activator of caspases) co-localizes with CGI-94 (comparative gene identification-94) in the cell. CGI-94 itself contains an utp11-motif and is probably involved in ribosome biogenesis and degenerative processes of AD. These findings point to a pivotal role of CGI-94 in the control of cellular survival.

Early postnatal application of thyroid hormones to rats results in morphological changes in septu... more Early postnatal application of thyroid hormones to rats results in morphological changes in septum and hippocampus. Modulation in the expression of either neurotrophins and/or their receptors is postulated to be responsible for these effects. In the present study we tested whether thyroxine administration leads to changes in the expression of neurotrophins of the nerve growth factor (NGF) family. Newborn rats were treated daily with subcutaneous injections of thyroxine until postnatal day (P) 12 at maximum. The pups were killed at defined intervals from P2 to 21. The septal area and the hippocampi were analyzed using the reverse transcriptase-PCR method for quantitation of NGF, brain-derived neurotrophic factor (BDNF), NT-3, and NT-4 messenger RNA (mRNA) levels. In hippocampus of hyperthyroid rats, as compared to controls, we found higher levels of BDNF and NT-3 mRNA over the total investigation period, whereas in the septum a thyroxine-dependent increase in NT-3 mRNA expression was observed. In addition, significant thyroxine-induced effects were found for all variables (except for NGF in the septum) at particular postnatal days. From these data we conclude that modulation of neurotrophin expression is a possible mechanism for the morphological modifications within the hippocampal mossy fiber system and the septohippocampal cholinergic system.

Early postnatal application of thyroid hormones to rats results in morphological changes of the s... more Early postnatal application of thyroid hormones to rats results in morphological changes of the septo-hippocampal cholinergic and the hippocampal mossy fiber systems. Modulation in the expression of either neurotrophins and/or their receptors is postulated to be involved in these effects. In a recent study, we showed that, after thyroxine application, the mRNA expression of neurotrophins of the nerve-growth-factor (NGF) family is significantly upregulated both in septum and hippocampus. To test whether the neurotrophin receptors (the low-affinity neurotrophin receptor p75 and the specific high-affinity receptors trkA, trkB, and trkC) were also affected by hormone administration, newborn rats were treated daily with subcutaneous injections of thyroxine until postnatal day 12 (P12) at latest. Control animals received corresponding injections of saline. The pups were sacrificed at defined intervals from P9 to P14. The septal areas and the hippocampi were analyzed using the reverse-transcription polymerase chain reaction (RT-PCR) method for quantification of p75, trkA, trkB, and trkC mRNA levels. Analysis of variance over the total investigation period revealed no significant general increases of the gene expressions of either neurotrophin receptor, neither in the septum nor in the hippocampus, although previous results have shown marked changes in neurotrophin levels. On particular postnatal days, significant upregulation could be observed in hippocampus for trkB and trkC. From these and recent data, we conclude that modulation of neurotrophin expression rather than neurotrophin-receptor expression contributes to the morphological modifications within the hippocampal mossy fiber system and the septo-hippocampal cholinergic system.