Conference

2018 ICAD abstracts

Cannabinoid receptor 1 antagonist (SR14176A) attenuates angiotensin II- induced premature senescence cellular in human coronary artery smooth muscle cells.

Al Abdullah W¹, Herbert K ¹, Lambert D ²

¹Department of Cardiovascular Sciences, University of Leicester, Cardiovascular Research Centre, Glenfield Hospital, UK.

²Department of Cardiovascular Sciences, University of Leicester, Division of Anaesthesia, Critical Care and Pain Management, UK.

Abstract:

Introduction: angiotensin II (Ang II) is associated with cardiovascular remodeling and ardiovascular disease via binding with its receptors Ang II type 1 receptor (hAT₁R). hAT₁R activation might induce vascular senescence that plays a critical role in age-related and vascular disease ^{(1, 2)}. It is essentially believed that senescence reflects some of the changes that occur through the aging of organisms, it detected in vivo of age related pathology such as atherosclerotic lesions⁽³⁾ and at sites predisposed to atheroma in man. Recent studies have demonstrated convincing of senescence occur in response to externally and internally induced stress signals in vivo⁽⁴⁾. Cellular senescence is a risk factor for progress of cardiovascular disease that include senescent vascular smooth muscle cells (VSMCs). However, cannabinoid receptor1 (hCB₁) antagonist utilise potent cytoprotective and anti-inflammatory effects in several preclinical disease models such as atherosclerosis⁽⁵⁾. Therefore, this project amid to investigate whether the possibility that hCB₁ antagonist could attenuates hAT₁R receptor mediated senescence of hCAVSMCs in vitro and whether the mechanism revers agonist of hCB₁ receptor SR141716A could have additional therapeutic value to prevent cellular senescence.

Methods: human coronary artery smooth muscle cells (hCAVSMCs) used from passage 1-6, incubated with SA- β-galactosidase staining solution for 24h in 37^°C without CO₂. SA- β-gal positive cells were detected under fluorescence microscopy. P53 and p21 express identified by immunoblotted assay.

Results: SA β-gal activity assessed in hCAVSMC cells after treated with Ang II over a range of concentrations (10^-12-10^-6 M) for 24 hours. Our data shows that Ang II caused a dose dependent increase in cell senescence with a maximal effect pEC ₅₀=8.98±0.08. Additional studies performed to investigate the reason of this rapid induction of premature senescence through pre-incubating hCAVSMCs cells with hCB₁ antagonist SR141716A (10µM) or hAT₁R antagonist E3174 (10µM), prior to challenge with Ang II (100nM) for 24h. The result show Ang II alone significantly increased SA β-gal stained cells, expression levels of P53 and P21 in hCAVSMCs. In contrast, cells were pre-treatment with SR141716A exhibited attenuation significantly of Ang II induced SA β-gal activity and expression of P53 and P21 (Figure1). Furthermore, E3174 (10µM) displayed a higher decrease and completely prevented Ang II-induced premature senescence.

Conclusion: the induction of premature senescence to be mainly hAT₁R mediated; however, it suggests that revers agonist of hCB₁ receptor regular VSMC senescence via mediating inactivation and/or blocking hAT₁R receptor signalling. It could be involved in modulate P53 or P21 pathway. Future work may include study activation hCB₁ for long time on hCASMC cells (Ministry of Higher Education and Scientific Research, Iraq, sponsors this study).

Key words: Cellular senescence, Angiotensin type 1 receptor (AT1R), CB1 antagonist SR141716A

References

1. Min LJ, Mogi M, Iwanami J, Li JM, Sakata A, Fujita T, et al. Cross-talk between aldosterone and angiotensin II in vascular smooth muscle cell senescence. Cardiovasc Res 2007; 76:506–16.

2.Kunieda T, Minamino T, Nishi J, Tateno K, Oyama T, Katsuno T, et al. Angiotensin II induces premature senescence of vascular smooth muscle cells and accelerates the development of atherosclerosis via a p21-dependent pathway. Circulation 2006; 114:953–60.

3. Vasile, E.; Tomita, Y.; Brown, LF.; Kocher, O. and Dvorak, HF. (2003). Differential expression of thymosin beta-10 by early passage and senescent vascular endothelium is modulated by VPF/VEGF: evidence for senescent endothelial cells in vivo at sites of atherosclerosis. FASEB J. 15:458-466.

4.Atyanarayana, S.U.; Wiemann, J.; Buer, J.; Lauber, K.E.J.; Dittmar, T.; Wustefeld, M.A.; Blasco, M.P.; Manns, and Rudolph, K.L. (2003). Telomere shortening impairs organ regeneration by inhibiting cell cycle re-entry of a subpopulation of cells. EMBO J. 22:4003-4013.

5.Sugamura, K.; Sugiyama, S.; Nozaki, T.; Matsuzawa, Y.; Izumiya, Y.; Miyata, K.; akayama, M.; Kaikita, K.; Obata, T.; Takeya, M. and Ogawa, H. (2009). Activated endocannabinoid system in coronary artery disease and anti-inflammatory effects of cannabinoid 1 receptor blockade on macrophages. Circulation, 119:28-36.

Differential Effects of Exosomes from Young and Old Rat Serum on Outcome after Ischemic Stroke in Aged Rats

Hongxia Zhang¹, Kunlin Jin

Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, TX76107, USA

Abstract:

Background: Aging is associated with striking increase in the incidence of stroke, which is the leading cause of disability and 5^th leading cause of death in the United States. Age also correlates with poor histology outcome and worsened neurobehavioral deficits after ischemic stroke. Our previous study evidenced that intraperitoneal administration of young plasma into aged ischemic rats significantly improved functional outcome and old plasma had verse effect. However, the underlying mechanisms remain largely unexplored. Here, we asked whether serum exosomes contribute to functional recovery after ischemic stroke.

Method: The exosomes were isolated from serum of young or old rats, and then were intravenously injected into aged ischemic rats for 3 days. Infarct volume was determined with TTC staining and motor function was determined with neurobehavioral tests.

Results: Our data showed that injection of serum exosomes derived from young rats into aged ischemic rats reduced infarct volume and improved functional impairments. On the contrary, injection of serum exosomes derived from aged rats increased infarct volume and worsened motor function.

Conclusion: Our data suggest that young and old serum exosomes differentiationally affect the ischemic stroke outcome.

Inferring Aging Regulation and Intervention from Big Data

Jing-Dong J. Han

Key Laboratory of Computational Biology, Chinese Academy of Sciences-Max Planck Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai, 200031, China.

ABSTRACT:

New high-throughput technologies, such as microarrays and deep sequencing technologies, have provided unprecedented opportunities for mapping mutations, transcripts, transcription factor binding and histone modifications at high resolution and at genome-wide level. This together with image-based phenome analysis has revolutionized the way regulations of diseases and other biological processes are studies and generated a large amount of heterogeneous data. We have explored to ab initio predict or reconstruct regulatory networks based on heterogeneous data on gene expression, histone modification and genomic changes and mapped various transcriptome and epigenome changes during aging and dietary interventions (Hou et al., 2016).

Recently by analyzing the 3D facial images, we generated the first comprehensive mapping of the aging human facial phenome. We found quantitative facial features, such as eye slopes, highly associated with age. We constructed a robust age predictor and found that on average people of the same chronological age differ by +/-6 years in facial age, with the deviations increasing after age 40. The predictor is as accurate as the most accurate to-date physiological age predictor – the one based on blood cell DNA methylation sites. Using this predictor we identified slow- and fast-agers that are significantly supported by health indicators (Chen et al., 2015). Based on our predicted outliers, we further profiled blood cell mRNA and lncRNA expression by RNA-seq and computationally predict their regulatory networks and their contributions to the difference in aging rate. By extending the study to a large Northern Chinese cohort of 10,000 people we can now use deep learning AI approaches to estimate health and aging status based on 3D facial images.

Astrocyte T-cell interaCtions Post-stroke: In vivo & In VITRO

Jessica Hersh, Luokun Xie, Wenjun Li, Ran Liu, and Shaohua Yang

University of North Texas Health Science Center at Fort Worth, Department of Neuroscience and Pharmacology UNT Health Science Center at Fort Worth, TX 76107, USA

ABSTRACT:

Post-ischemic stroke, T-lymphocytes enter the brain. The role of T-cells in the progression of cerebral infarction or repair mechanisms is unclear. We analyzed the function of T-cells in stroked regions by examining the pro- and anti-inflammatory interaction between T-cells and astrocytes.

In vivo, ischemic stroke was induced by middle cerebral artery occlusion in young adult C57/B6 male mice. Mice were sacrificed at 3 days or 1-month post-ischemic stroke. Paraffin-embedded brain sections demonstrated co-localization of astrocytes and CD4⁺ and CD8⁺T-cells in the ischemic region 1 month after stroke. T-cells were harvested from the brain by digestion; percoll enriched and incubated with anti-CD3 and CD25 antibodies. T-cells were sorted via flow cytometry. The cytokine profile of brain infiltrated CD4⁺ and CD8⁺ T-cells were compared to spleen T-cells using QT-PCR. The following cytokine gene expressions post stroke were found to be elevated: IFNγ, IL-10, IL-17, TNFα, and perforin.

In vitro, IL-10 gene expression was elevated in astrocytes and T-cells individually harvested from 1:1 co-culture compared to astrocytes and T-cells alone at 48 and 72 hours respectively. IL-10 was produced primarily by T-cells stimulated by direct contact with astrocytes. Anti-CTLA-4 antibodies block this direct cell-to-cell interaction by reducing IL-10 gene expression in both astrocytes and T-cells.

Our data suggests that T-cells release pro- and anti- inflammatory cytokines while in close proximity to astrocytes after ischemic stroke. In co-cultures, astrocytes directly interact with T-cells increasing their IL-10 gene expression by 72 h., implying a neuroprotective mechanism exists via astrocyte stimulation of T-cell IL-10 production.

The Association Between Gait Speed and Alzheimer’s Disease-vulnerable Cortical Thinning in Nondemented Older Adults

Seonjeong Byun¹, Hyang Jun Lee¹, Ji Won Han¹, Jun Sung Kim³, Euna Choi³, Ki Woong Kim^1,2,3,

¹Department of Neuropsychiatry, Seoul National University Bundang Hospital, Seongnam, Korea

²Department of Psychiatry, Seoul National University, College of Medicine, Seoul, Korea

³Department of Brain and Cognitive Science, Seoul National University College of Natural Sciences, Seoul, Korea

ABSTRACT:

Objectives: Slower gait speed predicts future risk of Alzheimer’s disease (AD). We investigated the association between gait speed and the magnetic resonance imaging derived thickness measurements across AD-vulnerable cortex in nondemented older adults.

Methods: We enrolled 98 community-dwelling nondemented elderly individuals aged 60 years or older (70.2 ± 7.0 years old, 49.0% women). We evaluated gait speed, cadence, and step length using a tri-axial accelerometer placed on the center of body mass and the GAITRite instrumented walkway as they walked at their comfortable speeds. We used FreeSurfer version 4.5 to process all MRI scans and computed average thickness values across AD-vulnerable cortical ROIs including the entorhinal cortex, temporopolar cortex, inferior parietal cortex, and posterior cingulate cortex from each hemisphere. Multivariable linear regression models were adjusted for total intracranial volume, age, sex, education level, and presence of the apolipoprotein E ε4 allele.

Results: In nondemented older adults, slower gait speed were associated with decreased average thickness in AD-specific cortex in both hemispheres after adjusting confounding factors. (beta=0.240, p=0.016 in left; beta=0.217, p=0.031 in right hemisphere) Among cadence and step length constituting gait speed, step length was associated with cortical thinning while cadence was not. (beta=0.213, p=0.030 in left; beta=0.221, p=0.025 in right hemisphere) In additional analysis to identify the association between each ROIs and gait parameters, enthorhinal and temporopolar cortical thickness exhibited a significant association with step length while inferior parietal and posterior cingulate regions did not. Posterior cingulate cortex thickness was associated with cadence.

Conclusion: Thinning in vulnerable cortical regions is the earliest cerebral structural imaging marker of AD. The observation that gait speed and step length are associated with thickness across AD-specific ROIs in nondemented older individuals suggests that the combination of those motor markers, neuroimaging, and laboratory markers may hold synergistic potential to aid in the prediction of AD.

Nucleophagy contributes to genomic stability

Muciño-Hernández Gabriel and Castro-Obregón Susana.

Department of Neurodevelopment and Physiology, Institute of Cellular Physiology, UNAM. Circuito Exterior S/N, Ciudad Universitaria, Coyoacán, 04510, Ciudad de México. Phone: (52)(55)5622-5676; e-mail: scastro@ifc.unam.mx

ABSTRACT:

Autophagy is a catabolic process which contributes to the cellular survival under stress conditions. From yeast to metazoans the genotoxic stress leads to the activation of this degradative process. However, autophagy seems to have more than one function as part of the DNA damage response (DDR). On one hand the repair of UV-induced DNA lesions or the proper DNA double strand breaks resolution requires autophagy. On the other hand, it could also contribute to the direct removal of damaged DNA, as micronuclei or DNA-protein crosslinks as Nucleophagy. Interestingly both genomic instability and autophagy disturbances occur during cancer development and aging. This way deepen in the study of autophagic degradation of nuclear damaged material contributes to understand the role of nucleophagy as a mechanism to maintain genomic stability. For that we have analyzed the process of damage and repair for DNA double strand breaks in MEF and A549 cancerous cells, in such a way that we detected the early and sustained activation of autophagy along the genotoxic stress and its resolution. In addition, we have been able to detect autophagic markers co-localizing with nuclear markers in alterations as micronuclei or nuclear blebbing suggesting the elimination of damaged material trough lysosomes. Finally, we have induced autophagy previous to DNA damage which surprisingly prevented DNA double-strand breaks in MEFs, while in A549 cells it had the opposite effect.

These results confirm the pro-survival role of autophagy by promoting genomic stability trough the degradation of nuclear damaged material.

Acknowledgements: Funding was provided by PAPIIT/UNAM IN206518, CONACyT CB2013-220515 and Fronteras 921 to SCO. CONACyT fellowship was awarded to GMH (417724). GMH is a student of the “Doctorado en Ciencias Bioquímicas, UNAM”. We acknowledge Dr. Beatriz Aguilar for her technical assistance.

The Association Between Gait Speed and Alzheimer’s Disease-vulnerable Cortical Thinning in Nondemented Older Adults

Seonjeong Byun¹, Hyang Jun Lee¹, Ji Won Han¹, Jun Sung Kim³, Euna Choi³, Ki Woong Kim^1,2,3,

¹Department of Neuropsychiatry, Seoul National University Bundang Hospital, Seongnam, Korea

²Department of Psychiatry, Seoul National University, College of Medicine, Seoul, Korea

³Department of Brain and Cognitive Science, Seoul National University College of Natural Sciences, Seoul, Korea

Abstract:

Effects of color and flickering frequency on gamma entrainment: a basic study for the therapeutics of Alzheimer’s disease

Kanghee Lee¹, Ye Seung Park², Seung Wan Suh¹, Ka-Young Tak², Ji Won Han¹, Jaeho Lee³, Jaehyeok Park³, Seunghyup Yoo³, Ki Woong Kim^1,2,4

¹Department of Neuropsychiatry, Seoul National University Bundang Hospital. Seongnam, 13620, Republic of Korea

²Department of Brain and Cognitive Science, Seoul National University, Seoul, 08826, Republic of Korea.

³School of Electrical Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Republic of Korea.

⁴Department of Psychiatry, Seoul National University, College of Medicine, Seoul, 03080, Republic of Korea

Abstract:

Brain photobiomodulation (PBM) is getting attention as a novel therapy for Alzheimer’s disease (AD). Gamma oscillation entrained by 40Hz flickering light was known to lead to a decreased amyloid β production and an increased amyloid endocytosis by microglial activities in the visual cortex of mouse model. To apply gamma entrainment to the therapeutics for AD, this study aims to find the optimal conditions of light stimuli including colors and flickering frequencies for entraining gamma oscillations in the brains of healthy adults. Eighteen participants put on the goggles during the electroencephalography (EEG) recording which contains a pair of organic light emitting diodes (OLED) panels. They were instructed to keep eyes forward to panels which flickered at four different colors (white, red, green, or blue) and ten different frequencies (32-50Hz) in 10 cd/㎡ luminance intensity. We found that steady-state visual evoked potentials (SSVEPs) under the white or red light was higher than those under the green or blue with slowly flickering frequencies (<38Hz) at Oz. The SSVEPs under white or red light was also higher than those under green or blue light in the parietal sites. A strong negative correlation between flickering frequency and relative power change at that frequency was also observed. These results imply that flickering light with lower gamma in white or red is efficient to induce the synchronization of brain activity. Entrainment of gamma EEG activity may take a role for restoration of cognitive function. This study takes the first step to develop a PBM therapeutics for AD.

Effect of hypotension on the risk of cerebral white matter hyperintensities in the elderly

Jun Sung Kim¹, Subin Lee¹, Jae Hyoung Kim³, Ji Won Han⁴, Jong Bin Bae⁴, Seung Wan Suh⁴, Ji Hyun Han⁴, Ki Woong Kim^1,2,4,*

¹Department of Brain and Cognitive Science, Seoul National University College of Natural Sciences, Seoul, South Korea

²Department of Psychiatry, Seoul National University, College of Medicine, Seoul, South Korea

³Department of Radiology, Seoul National University Bundang Hospital, Seongnam, South Korea

⁴Department of Neuropsychiatry, Seoul National University Bundang Hospital, Seongnam, South Korea

Abstract:

White matter hyperintensity (WMH) may have different etiologies by their locations. Smooth juxtaventricular WMH (JVWMH) possibly linked to the increase of interstitial fluid (cerebrospinal fluid leakage), irregular periventricular WMH (PVWMH) are likely determined by chronic hemodynamic insufficiency (hypoperfusion), whereas deep WMH (DWMH) may be attributed to small vessel disease. In this study, we investigated the effects of chronic low blood pressure (BP) on the volume of cerebral WMH. The study included 475 cognitively normal elderly subjects who participated in the Korean Longitudinal Study on Cognitive Aging and Dementia (KLOSCAD). We measured BP using automatic sphygmomanometer. From 3.0T FLAIR MR images, we segmented WMH and classified them into JVWMH, PVWMH and DWMH automatically using in-house program previously reported. In the linear regression models, the volume of PVWMH was associated with systolic BP (F = 4.723, p = 0.045) but not with diastolic BP (F = 4.723, p = 0.831). Neither JVWMH nor DWMH was associated with systolic and diastolic BPs. The participants whose systolic BP was 110 mmHg or below had more PVWMH than those whose systolic BP was over 110 mmHg (F=6.059, p=0.014). This was the case in the subjects who had controlled hypertension (F=6.387, p=0.012) but not in those who did not have hypertension (F=1.167, p=0.281). In conclusion, low systolic BP may be a risk factor of PVWMH in controlled hypertension patients.

Synergistic and antagonistic interventions between longevity genes - the SynergyAge database

BUNU Gabriela¹, ION Cătălin-Florentin¹, TĂCUTU Robi¹

¹Institute of Biochemistry of the Romanian Academy, Bucharest, Romania

Introduction: The aging process can be modulated using different approaches with the most extraordinary genetic interventions in model organisms increasing lifespan up to 10x (in C. elegans). Currently, there are more than 2000 known longevity-associated genes (LAGs), whose knockout or overexpression result in a long- or short-lived phenotype. However, when combining two or more genetic interventions, the effect is rarely additive, as genes can be epistatic and interact in nonlinear ways. In order to evaluate the synergism and antagonism of LAGs, we are developing a database with gene combinations that affect lifespan.

Methods: Data is manually curated from scientific articles with experimentally validated results, describing at least one long- or short-lived genetic model with at least two interventions (mutations, knockout, overexpression or RNA interference). We include only experiments in which the “multiple” mutant is compared with single gene mutants and, if possible, with wild type. Only articles which include survival curves characteristic for a normal aging pattern are selected. Besides lifespan values, we also record other factors (e.g.: temperature, diet, etc). We classify interactions as synergistic (independent), almost additive, dependent or antagonistic.

Results: So far, the database contains almost 600 combinations of genes and 1700 lifespan values for C. elegans, D. melanogaster and M. musculus. Data will be available for visualization through an easy to use web interface. This work is supported by the Competitiveness Operational Programme 2014-2020, POC-A.1-A.1.1.4-E-2015

Auditory guidance of imagined movements: Effects of real-time auditory feedback (sonification) guided mental imagery on knee proprioception

Shashank Ghai*, ¹Gerd Schmitz, ¹Tong-Hun Hwang, ¹Alfred O. Effenberg

Abstract:

Proprioception is an integral prophylactic component for executing motor tasks effectively across all age groups. Recently, motor imagery and augmented sensory feedback have garnered a lot of attention in rehabilitation interventions to facilitate proprioception. In this pilot study, we aimed to analyze the influence of real-time auditory feedback and mental imagery on proprioceptive learning. 42 healthy participants were randomly allocated into three groups. All the groups initially trained bilaterally at the knee joints with real-time auditory feedback for four target angles (20°, 40°, 60° and 80°). Thereafter, the groups continued the training with/without mental imagery, and with/without auditory guided mental imagery. During mental imagery condition the participants were verbally instructed to imagine knee re-positioning at the trained four angles upon verbal instructions. Retention measurements were performed after 15 minutes and 24 hours of the final proprioceptive test. Likewise, a generalized knee-proprioceptive test assessed the performance on four untrained angles (10º, 30º, 50º and 70º). Statistical analysis revealed significant enhancement in proprioceptive accuracy for the auditory guided mental imagery group as compared to group performing mental imagery without auditory guidance, and group performing no mental imagery. We also observed significant enhancement in generalized knee-proprioception on four untrained angles for all the groups. In addition, significant enhancement in self-reported attention was observed for auditory guided mental imagery group as compared to conventional mental imagery group. This study, for the first time demonstrates beneficial effects of auditory guided mental imagery on knee proprioception and provides evidence of intermodal learning during imagination of movements. The data from this study will support the development of future clinical trials where similar proprioceptive parameters will be evaluated in elderly fall prone population groups. This trial was registered in the German Clinical Trial Registry DRKS00014244.

Key words: Perception, imagery, rehabilitation, sonification, coordination, joint position sense, motor learning

Necrostatin-1 prevents necroptosis in brains after ischemic stroke via inhibition of RIPK1-mediated RIPK3/MLKL signaling

Feng-Yan SUN, Xu-Xu Deng and Shan-Shan Li,

Department of Neurobiology, School of Basic Medical Sciences，Institute for Basic Research on Aging and Medicine, and State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China, fysun@shmu.edu.cn

Abstract:

Pharmacological studies have indirectly shown that necroptosis participates in ischemic neuronal death. However, its mechanism has yet to be elucidated in the ischemic brain. TNFα-triggered RIPK1 kinase activation could initiate RIPK3/MLKL-mediated necroptosis under inhibition of caspase-8. In the present study, we performed middle cerebral artery occlusion (MCAO) to induce cerebral ischemia in rats and used immunoblotting and immunostaining combined with pharmacological analysis to study the mechanism of necroptosis in ischemic brains. In the ipsilateral hemisphere, we found that ischemia induced the increase of (i) RIPK1 phosphorylation at the Ser166 residue (p-RIPK1), representing active RIPK1 kinase and (ii) the number of cells that were double stained with P-RIPK1 (Ser166) (p-RIPK1⁺) and TUNEL, a label of DNA double-strand breaks, indicating cell death. Furthermore, ischemia induced activation of downstream signaling factors of RIPK1, RIPK3 and MLKL, as well as the formation of mature interleukin-1β (IL-1β). Treatment with necrostatin-1 (Nec-1), an inhibitor of necroptosis, significantly decreased ischemia-induced increase of p-RIPK1 expression and p-RIPK1⁺ neurons, which showed protection from brain damage. Meanwhile, Nec-1 reduced RIPK3, MLKL and p-MLKL expression levels and mature IL-1β formation in Nec-1 treated ischemic brains. Our results clearly demonstrated that phosphorylation of RIPK1 at the Ser166 residue was involved in the pathogenesis of necroptosis in the brains after ischemic injury. Nec-1 treatment protected brains against ischemic necroptosis by reducing the activation of RIPK1 and inhibiting its downstream signaling pathways. These results provide direct in vivo evidence that phosphorylated RIPK1 (Ser 166) plays an important role in the initiation of RIPK3/MLKL-dependent necroptosis in the pathogenesis of ischemic stroke in the rodent brain.

Keywords: RIPK1; necroptosis; cerebral stroke;

The Effect of Aquaporin-4 Knockout on Interstitial Fluid Flow and the Structure of the Extracellular Space in the Deep Brain

Aibo Wang^1,2, Yuanyuan Li^1,2, Ze Teng^1,2, Peng Wang³, Rui Wang2, Wei Wang2, Hongbin Han^{1,2, *}

¹Department of Radiology, Peking University Third Hospital, Beijing 100191, China ²Beijing Key Lab. of Magnetic Resonance Imaging Technology, Beijing 100191, China ³Department of Orthopedics, Peking University Third Hospital, Beijing 100191, China

Abstract:

It has been reported that aquaporin-4 (AQP4) deficiency impairs transportation between the cerebrospinal fluid and interstitial fluid (ISF) as well as the clearance of interstitial solutes in the superficial brain area. However, the effect of AQP4 on ISF flow in the deep brain remains unclear. This study compared the brain ISF flow in the caudate nucleus and thalamus of normal rats (NO) and AQP4 knockout rats (KO) using tracer-based magnetic resonance imaging. The rate of brain ISF flow slowed to different degrees in the two regions of KO rats’ brains. Compared with NO rats, the half-life of ISF in the thalamus of KO rats was significantly prolonged, with a corresponding decrease in the clearance coefficient. The tortuosity of the brain extracellular space (ECS) was unchanged in the thalamus of KO rats. In the caudate nucleus of KO rats, the volume fraction of the ECS and the diffusion coefficient were increased, with significantly decreased tortuosity; no significant changes in brain ISF flow were demonstrated. Combined with a change in the expression of glial fibrillary acidic protein and AQP4 in two brain regions, we found that the effect of AQP4 knockout on ISF flow and ECS structure in these two regions differed. This difference may be related to the distribution of astrocytes and the extent of AQP4 decline. This study provides evidence for the involvement of AQP4 in ISF transportation in the deep brain and provides a basis for the establishment of a pharmacokinetic model of the brain’s interstitial pathway.

Experimental DNA damage induces IL-18 release and NLRP3-inflammasome-dependent IL-1 production in human retinal pigment epithelium cells

Korhonen Eveliina⁽¹, Piippo Niina⁽¹, Kaarniranta Kai^(2,3, Hytti Maria⁽¹, Kauppinen, Anu⁽¹

¹School of Pharmacy, University of Eastern Finland, Kuopio, Finland

²Department of Ophthalmology, Kuopio University Hospital, Kuopio, Finland

³Department of Ophthalmology, University of Eastern Finland, Kuopio, Finland

Abstract:

DNA damages accumulate in aged cells but their role in age-related diseases has remained elusive. Here, we have induced DNA damage by ultraviolet B (UVB) irradiation in IL-1-primed human retinal pigment epithelium (ARPE-19) cells and examined the subsequent inflammatory response. For studying mechanisms in more detail, cells were pre-treated with potassium chloride, n-acetyl-cysteine (NAC), or mitoTEMPO. Pro-forms of IL-1 and IL-18 were measured by qRT-PCR, and secreted inflammasome-related proteins IL-1β, IL-18, and caspase-1 using the enzyme-linked immunosorbent assay (ELISA). NLRP3 was also detected from the culture medium samples by ELISA and additionally, it was blocked using a specific siRNA. Cell-based ELISA was used to detect cyclobutane pyrimidine dimers (CPDs), ROS levels were measured by the DCFDA assay, and the levels of extracellular ATP were determined using a commercial assay. IL-1 priming induced the expression of pro-IL-1 but pro-IL-18 was constitutively expressed in ARPE-19 cells. Both cytokines were released from cells following the UVB exposure but NLRP3 was involved only in the production of IL-1. Potassium efflux contributed to the release of IL-1, whereas ROS participated in the secretion of IL-18. Our results indicate that one stress factor can induce distinct pathways for producing inflammasome-related cytokines IL-1 and IL-18 in human retinal pigment epithelium cells.

Association between inflammation, oxidative stress and cellular ageing in infection: Experimental human malaria challenge

Authors:

Aurelie Miglar1, Isaie Reuling2, Anna Färnert1,3, Robert Saeurwein2, Muhammad Asghar1*

1Division of Infectious Diseases, Department of Medicine Solna, Karolinska Institutet, SE-171

76 Stockholm, Sweden.

2Department of Medical Microbiology, Radboud University Medical Center, Geert Grooteplein

28, Microbiology 268, 6500 HB, Nijmegen, The Netherlands.

3Department of Infectious Diseases, Karolinska University Hospital, Stockholm, Sweden.

Abstract:

Background Infectious diseases can potentially affect cellular ageing by adding miles to the biological clock. Accelerated cellular ageing and reduced lifespan have been linked to chronic asymptomatic malaria infection in birds and single acute malaria infections have been shown to affect cellular ageing in travellers. In this controlled human malaria challenge study, we confirm the effect of acute malaria infection on cellular ageing and reveal the underlying mechanisms leading to this effect. Methods In an open-label randomized trial, healthy, malaria naïve individuals were infected with the malaria parasite Plasmodium falciparum, administered antimalarial treatment, and followed until day 64 post-infection. Telomere length was measured by real-time quantitative PCR (qPCR), whereas telomerase and cycling dependent kinase inhibitor 2A (CDKN2A) expression, oxidative stress levels were evaluated by reverse transcriptase PCR (RT-qPCR), and cytokines by Luminex technology. Results Our findings provide evidence that malaria infection affects cellular ageing which is linked to parasite burden in the host, reflected by telomere shortening and increased CDKN2A levels at day 12 post malaria challenge. Furthermore, after successful treatment the effect was reversed again, confirming that the effect was linked to the infection with malaria parasites. Telomere length and CDKN2A expression were significantly correlated with oxidative stress markers, inflammatory cytokines and liver function test abnormality, suggesting that even a short exposure to the malaria parasite affects cellular ageing by elevating the inflammatory cytokines production and disturbing the oxidative stress balance. In conclusion, our findings demonstrate underlying cellular mechanisms by which malaria infection affects cellular ageing.

Efficacy and safety of sequential alprostadil therapy in cerebral small vessel disease: A prospective randomized multicenter study

Wenyuan. Tao, Yun Xu

Drum Tower Hospital- Medical School of Nanjing University, Department of Neurology.

Abstract:

Objective: The role of alprostadil in cerebral small vessel disease (CSVD) still remains controversial. The purpose of this study was to evaluate the efficacy and safety of sequential alprostadil therapy in patients with CSVD.

Methods: This was a multicenter, prospective, randomized, controlled trial. 119 CSVD subjects from 25 centers were randomly assigned to aspirin group (n = 60) and alprostadil + aspirin group (n = 59). Both groups received 100 mg per day aspirin for 1 year, and the alprostadil + aspirin group additionally received 20 ug per day alprostadil for 10 days followed by 120ug per day for 3 months. The primary outcome were the risk of ischemic stroke and hemorrhagic events over the 1-year treatment period. Cognitive function, gait and brain MRI data were also assessed.

Results: No significant difference was found in the baseline characteristics between the two groups. At the end of follow-up, the risk of ischemic stroke was significantly lower in the alprostadil + aspirin group than in the aspirin group (relative risk, 0.74 [95% CI, 0.65 to 0.88]), controlling for demographic characteristics and vascular risk factors. In contrast, the risk for hemorrhagic events was similar between the two groups (relative risk, 0.92 [95% CI, 0.79 to 1.13]). In addition, smaller volume and a smaller number of lacunar infarction were found in the alprostadil + aspirin group (P = 0.022 and P = 0.045 respectively). Although no significant differences of cognitive function and gait performance were shown between groups, the alprostadil + aspirin group displayed a slower rate of decline in MoCA scores (P = 0.047).

Conclusion: Sequential alprostadil therapy is a safe and effective treatment for reducing the risk of ischemic stroke in CSVD subjects. These results provide further evidence that alprostadil probably has a therapeutic effect in the treatment of CSVD.

The relationship between white matter hyperintensities volume and cognitive function in elderly patients with hypertension

Lili Zhai and Yun Xu

Department of Neurology, Drum Tower Hospital, Medical School of Nanjing University, Nanjing 210008, Jiangsu, China

Abstract:

Objective: The aim of this study was to explore the correlation between white matter hyperintensities volume and cognitive function in elderly patients with hypertension. Methods: 82 elderly patients with hypertension from January 2017 to December 2017 were recruited in this study sequentially, aged between 50 and 80 years. All subjects underwent 3.0 T magnetic resonance imaging and used automatic white matter quantitative technique to get their personal white matter hyperintensities total volume, periventricular white matter hyperintensities volume and deep white matter hyperintensities volume. Neuropsychological scales were used to evaluate their global cognitive function, memory function, fluency, executive function and processing speed. We assessed the relationship between white matter hyperintensities volume and cognitive performance via linear regression analysis. Results: After adjusting for age, sex and education, Victorial Stroop Test-(2-1) which reflect executive function was related to all the position of white matter hyperintensities volume (P<0.05), Victorial Stroop Test-2 which reflects processing speed was related to periventricular white matter hyperintensities volume (Standardized β-values=0.228 P<0.05), after additional adjusting for systolic blood pressure and years of hypertension, these results still existed. Conclusion: Whole white matter hyperintensities volume，periventricular white matter hyperintensities volume and deep white matter hyperintensities volume are related to executive function, periventricular white matter hyperintensities volume is also related to processing speed in patients with hypertension, and the relationship between white matter hyperintensities volume and cognitive domain don’t influenced by blood pressure control state and hypertension years.

Functional network efficiency driving the association between cerebral small vessel disease and cognitive performance

Hui Zhao and Yun Xu

Department of Neurology, Drum Tower Hospital, Medical School of Nanjing University, Nanjing 210008, Jiangsu, China

BACKGROUND:

Cerebral small vessel disease (SVD) is a major cause of cognitive decline in the elderly. However, the topological network alterations in SVD with cognitive impairment remained unclear. We aimed to investigate how these changes in functional network were related to cognitive decline and whether they could be used in discriminant analysis for identifying SVD individuals with cognitive impairment.

METHODS:

We recruited 36 SVD with normal cognition (SVD-NC), 38 SVD with cognitive impairment (SVD-CI) and 38 healthy control (HC) individuals. The functional network was constructed from resting-state functional magnetic resonance imaging. The six subnetworks (default mode network; cingulo-opercular network; fronto-parietal network; occipital network; sensorimotor network; and cerebellum network) were obtained based on Dosenbach atlas. The measures of efficiency using graph theory were calculated. Linear regression analyses and mediation analyses were computed to assess the relationship between SVD burden, network measures and neurocognitive performance. The functional network measures were subsequently applied to discriminant analysis using a support vector machine (SVM) classification method.

RESULTS:

We found that global and local efficiency of the whole brain were significantly decreased in SVD-CI. The subnetwork analysis indicated that SVD-CI showed reduced functional efficiency, particularly in fronto-parietal network and cingulo-opercular network. Reduced nodal efficiency were locally significant in prefrontal, parietal and anterior cingulate cortex regions in SVD-CI. In multiple regressions models, nodal efficiency of prefrontal and parietal regions were related to information processing speed function and mediated the associations between periventricular white matter hyperintensity and cognition. Using the SVM algorithm, SVD-CI was differentiated from SVD-NC using the selected functional efficiency measures and the accuracy reached up to approximately 82% with leave-one-out cross-validation during the discrimination process.

CONCLUSION:

Our findings provide evidence that functional network efficiency might mediate the association between SVD and cognitive performance. The classification results indicate that network efficiency measures might provide an useful tool as disease marker.

KEY WORDS:

cerebral small vessel disease, graph theory, network efficiency, support vector machine

The resting-state functional pattern of parietal cortex in progressive cognitive impairment of cerebral small vessel disease

Ying Liu and Yun Xu

Department of Neurology, Drum Tower Hospital, Medical School of Nanjing University, Nanjing 210008, Jiangsu, China

Abstract:

BACKGROUND:

Progressive cognitive impairment (CI) is a common symptom in cerebral small vessel disease (CSVD). Emerging evidence has suggested that CI is associated with altered brain function. However, there is limited knowledge regarding the changing patterns of brain function in CSVD patients with progressive CI.

METHODS:

Ninety-four subjects were recruited in this study, including 29 non-CSVD subjects with normal cognition (NC), non-CSVD subjects with CI, 24 CSVD subjects with NC and 24 CSVD subjects with CI. All participants underwent multimodal magnetic resonance imaging (MRI) and a series of neuropsychological tests. The amplitude of low-frequency fluctuation (ALFF) was used to detect cerebral spontaneous activity. Moreover, the numbers of lacunes, perivascular spaces (PVs) and cerebral microbleeds (CMBs) were evaluated, and the volume of white matter hyperintensities (WMH) was automatically calculated with the Wisconsin WMH Segmentation Toolbox. The parietal cortex (i.e., the inferior parietal lobule and precuneus) is vulnerable to the interactive effects of disease status (non-CSVD versus CSVD) and cognitive status (NC versus CI).

RESULTS:

Post hoc analysis indicated that these interactive regions showed an inverse U-shaped pattern in these four groups: more brain activity was exhibited by the CSVD with NC subjects than the non-CSVD groups, followed by decreased activation with the progression of CI in the CSVD with CI subjects. Interestingly, the present study further found the lacunar numbers may participate in the regulation of brain function in CSVD with CI subjects.

CONCLUSION:

These findings suggest that dynamic changes to the inferior parietal lobule and precuneus may be considered neuroimaging biomarkers that indicate CSVD progression. Our results also indicate the vital role of lacunes in brain function and that anti-lacune therapy may slow the advancement of CI in CSVD subjects.

Molecular Mechanisms influencing the Cytokine Inflammation Pathway in Healthy Ageing and Age-related diseases

Irene Maeve Rea, Queens University, Belfast

The inflammatory response must be tightly controlled to ensure effective immune protection thorough out life: otherwise uncontrolled inflammation can become damaging and destructive. Cytokine dysregulation is believed to play a key role in the remodeling of the immune system at older age, where loss of the fine-control of systemic inflammation seems to be associated with both good quality ageing and age-related disease. The reshaping of the cytokine expression pattern, with a progressive tendency toward a pro-inflammatory phenotype has been called ‘inflamm-aging’ and seems to associate with most major age-related diseases such as atherosclerosis, diabetes, rheumatoid arthritis and ageing itself.

Several common molecular pathways have been associated with low-grade inflammation, and activation of the NF-kB cytokine and the IL-1b-mediated inflammatory cascade. The age-related change in redox balance, the increase in age-related senescent cells and senescence-associated secretory phenotype (SASP), and the decline in effective autophagy can trigger the inflammasome, which is central to the inflammatory cascade. Here we will discuss some aspects of the present understanding of the molecular mechanisms that trigger inflammation and related it to findings in various age-related diseases and from long-lived nonagenarian cohorts who exemplify good quality ageing.

Key words: “Age-related diseases”, “Healthy Ageing”, “Inflammatory Triggers”

Sex Difference in the Effect of Aging on Sleep: A Prospective Elderly Cohort Study

Seung Wan Suh, Ki Woong Kim

Seoul National University Bundang Hospital, 82, Gumi-ro 173beon-gil, Bundang-gu, Seongnamsi, Gyeonggi-do, Republic of Korea.

ABSTRACT:

OBJECTIVES: To present normative sleep parameters in Korean elderly and to examine the sex difference in the effect of aging on these parameters.

METHODS: We gathered data from the Korean Longitudinal Study on Cognitive Aging and Dementia (KLOSCAD) cohort (N = 2,222, mean age 69), composing of community-dwelling elderly aged 60 or older. Exclusion criteria included those with any serious psychiatric/neurologic disorders and sleeping pill users. Sleep parameters and other clinically relevant information were obtained at baseline, and after 4 years of follow-up, using Pittsburgh Sleep Quality Index. The effect of physiologic aging was analyzed using repeated measure analysis of variance (RM-ANOVA) in male and female groups respectively, by each of the age tertiles, adjusted for the type of residential area, presence of cohabitant, smoking/alcohol/coffee consumption status, and physical activity.

RESULTS: We observed that the normative value of sleep duration, bedtime, wake time, sleep latency, and sleep efficiency are 388 (76) mins, 22:30 (00:18), 05:42 (00:17), 22 (25) mins, and 90 (12) % at baseline, respectively (mean [SD]). After 4 years, we found that female participants showed a significant increase in the scores of subjective sleep quality and advance in bedtime, while male participants showed no significant findings.

CONCLUSION: Unlike previously reported cross-sectional data, physiologic aging may have different effect on each of sleep parameters according to sex. Therefore, clinicians should be cautious when determining whether a patient has a pathologic sleep-wake cycle or not.

Key Words: Ageing, Sleep, Sex difference, Normative value, Observational Study

Sequential Sleep Architecture May Predict Cognitive Decline: A Prospective Cohort Study

Ki Woong Kim

Seoul National University Bundang Hospital, 82, Gumi-ro 173beon-gil, Bundang-gu, Seongnamsi, Gyeonggi-do, Republic of Korea.

ABSTRACT:

Study Objectives: To investigate the effect of successive NREM/REM cycles and their associated sleep architecture on incident cognitive decline.

Methods: Data were acquired from a subset of the Korean Longitudinal Study on Cognitive Ageing and Dementia (KLOSCAD) cohort composed of cognitively normal elderly aged 60 or older who underwent an overnight polysomnography at baseline. A NREM/REM cycle was defined as a sequence of NREM and REM sleep, uninterrupted by more than 2 minutes of a waking period. After 4 years, cognitive status was evaluated and related to measures of sleep architecture including cycle-related parameters by logistic regression analyses.

Results: Of the 235 cognitively normal subjects (mean [SD] age 68 [5]; 60% female) at baseline, 14 (5.9%) demonstrated cognitive decline (becoming mild cognitive impairment or dementia) at follow-up. The number of cycles (odds ratio [OR], 1.63 [95% CI, 1.09–2.43]; p=.016) and average cycle time (OR, 0.97 [95% CI, 0.94–0.99]; p=.014), when analyzed separately, were significantly associated with cognitive decline. When sleep period time, total cycle time, and the number of cycles were analyzed simultaneously, total cycle time (OR, 0.99 [95% CI, 0.98–0.99]; p=.034) and the number of cycles (OR, 2.35 [95% CI, 1.33–4.15]; p=.003) were found to be significantly related to cognitive decline.

Conclusions: Short and fragmented NREM/REM cycles in cognitively normal elderly may be early markers of incident cognitive decline.

Impaired activation of oval cells is associated with malfunction of hepatic stellate cells in aged mice

Qiwei Wang¹, Liu Qian¹, Yanan Wang¹and Yanyun Zhang¹

¹Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

Abstract:

Aging predisposes to structural and functional impairment of liver which is associated with increased risk of hepatic injury. Recently tissue specific stem cells are reported to participated in the regulation of age-induced organism injury. In this study, taking advantage of well-established murine oval cells (OC) activation model, we demonstrated that OC was less activated with aging, which was mainly due to malfunction of the niche. We found that laminin which mainly produced by hepatic stellate cells was lower in aged mice tissues than that in young liver tissues constructing a “senescent” OC niche resulting in the decrease regenerative ability of OC delaying the restoration of liver function.

Activation of central neuropeptide Y receptors counteracts spatial memory impairments associated with ageing and amyloid-β oligomers (AβOs)

Fernanda Gomes de Queiroz Barros-Aragão^1,2, Ana Cristina Guerra de Souza², Jessica Barbosa³, Marissa Schamne², Angela Patricia França², Sérgio Teixeira Ferreira^1,4, Rui Daniel Prediger², Claudia Pinto Figueiredo^1,3

¹Programa de Ciências Morfológicas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro

²Programa de Pós-Graduação em Farmacologia, Universidade Federal de Santa Catarina

³Faculdade de Farmácia, Universidade Federal do Rio de Janeiro

⁴Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro

Abstract:

Several evidences indicate that the neuropeptide Y (NPY) signaling is compromised in the central nervous system during ageing and Alzheimer’s disease (AD). Here we evaluated the effects of central administration of NPY Y receptors agonists over the cognitive deficits associated to ageing or to amyloid-β oligomers (AβOs) administration in mice. Moreover, the effects of NPY and NPY Y2 receptor selective agonist NPY (13-36) over the reactive oxygen species (ROS) production induced by AβO were evaluated in a neuronal hippocampi cell culture. A single NPY (23,4 pmol) intracerebroventricular (i.c.v.) injection reversed age-related spatial memory deficits in 12-month-old C57BL/6 mice addressed in the object location test. Pre-treatment with NPY or the Y2 receptor agonist NPY(13-36) (23,4 pmol, i.c.v.) prevented spatial memory deficits induced by AβOs (10 pmol, i.c.v.) in the object location test in 3-month old Swiss mice. Moreover, treatment with NPY(13-36) prevented ROS generation induced by AβOs (0,5μM) in vitro. None of the tested compounds significantly altered the behavioral parameters related to locomotion or emotionality. These results support the hypothesis that NPY signaling restoration and the use of Y2 receptor agonists are potential strategies for the management of age-associated cognitive deficits as well as those present in initial stages of AD.

Keywords: Alzheimer’s Disease, Ageing, Neuropeptide Y2 Receptor.

Tuberculosis in Elderly: A spatial temporal analysis using a Conditional auto-regressive model in Mainland China

Endawoke Amsalu^{1, 2}, Mengyang Liu^{1, 2}, Qihuan Li^{1, 2}, Xiaonan Wang^{1, 2}, Lixin Tao^{1, 2}, Xiangtong Liu^1,2, Yanxia Luo^1,2, Xinghua Yang^1,2, Yingjie Zhang³, Weimin Li⁴, Xia Li⁵, Wei Wang ⁶, Xiuhua Guo^{1, 2*}

¹Department Epidemiology and Health Statistics, School of Public Health, Capital Medical University, Beijing 100069, China;

²Beijing Municipal Key Laboratory of Clinical Epidemiology, Beijing 100069, China;

³Chinese Center for Disease Control and Prevention, Beijing 102206, China;

⁴Beijing Chest Hospital, Beijing 101149, China

⁵Department of Mathematics and Statistics, La Tribe University, Melbourne, 3086, Australia

⁶Global Health and Genomics, School of Medical Sciences and Health, Edith Cowan University,

Joondalup, Perth, WA6027, Australia

Abstract:

Background: Tuberculosis (TB) morbidity and mortality in the elderly is an increasing problem in many countries. Understanding of geographical variation of disease especially with respect to climate factors at the small-area scale in the specific group is necessary. We examined spatial-temporal pattern TB in Elderly and its relation with meteorological & socio demographic factors using Bayesian conditional autoregressive (CAR) model.

Method: An Ecological design was conducted in elderly (age >=65) using Chinese Centre for Disease Prevention and Control (CCDC) surveillance database from 2005-2015. Spatial auto-correlation and hot map was explored using geographical information system (GIS) statistics. Bayesian CAR model using Markov chain Monte Carlo (MCMC) simulation with Gibbs sampling technique was used for modelling TB disease and to estimate the parameter in WinBUGS software. Deviance information criteria (DIC) used to select the best performing model.

Results: Spatially, TB in elderly was clustered in Central and South-East China. Temporally, an increasing trend and high peak of TB in spring was detected. Bayesian CAR model revealed that spatial-clustering of TB in elderly was significantly associated with temperature of posterior mean value: -0.165(95%CI: -0.235, -0.108) and average wind: -0.028 (95%CI: -0.043, -0.018) negatively, and with rainfall 0.095(95%CI: 0.045, 0.163) positively. Relative humidity was not significant. TB in elderly was significantly and positively associated with population density 0.088(95%CI: 0.031, 0.129) and sex ratio (M: F) with 0.162 (95%CI: 0.091, 0.284), and negatively related with growth domestic product (GDP) with: -0.046(95%CI: -0.156, -0.037). Out of 31 provinces, 17 provinces had the higher risk of tuberculosis.

Conclusion: TB in elderly shows a clear spatial variation and geographically aggregation. TB in elderly is a seasonal disease and men are more affected. The elderly populations in areas with underprivileged economy, high population density, high rainfall, week wind speed, and low temperature have higher risk for TB.

Keywords: TB, Bayesian, Spatial, Spatial temporal, Elderly

Melatonin inhibits inflammasome mediated pyroptosis and improves functional outcome after mild traumatic brain injury

Felix Siaw-Debrah^1,2*, Mark Nyanzu^1,2*, Pan Sishi^1,2, He Zibin^1,2, Jin Kunlin², Zhuge Qichuan^1,2, Huang Lijie^1,2

1 Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China;

2 Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China

*Co-authors contributed equally to this study.

Abstract:

Traumatic brain injury (TBI) refers to injury to the brain due to penetrating object or blow causing dysfunction of the brain, resulting in either temporary or permanent impairment of neurological function. TBI episode results in the activation of inflammasomes mediated pathways, as well as other cellular death pathways. These pathways help in the removal of inactivated and damaged cells but could also result in nerve system damage. Pyroptosis, a newly discovered cell death pattern has received major attention in recent years. It mainly depends on Caspase-1-mediated pathways to cause cell death. Our research focused on the effect of melatonin, a known neuroprotective agent, on inflammasomes mediated pyroptosis. We examined the effect of melatonin on the activity of inflammasome complex NLRP3, ASC, and procaspase BV2 cells and in mild TBI mice model. LPS was used to induced inflammation in murine microglia cell (bv2) TBI by cortical contusion injury (CCI) and discovered that the administration of melatonin((2.5mg/100g) 1ml/100g) could reduce caspase-1 activation, NLRP3 inflammasome activation and inhibit IL-1β production and maturation. We conclude that melatonin attenuates pyroptosis and improves functional outcome and therefore warrant further studies for the treatment of TBI.

Keywords：Traumatic brain injury; Pyroptosis; Caspase-1; IL-1β; microglia

Resveratrol attenuates endoplasmic reticulum stress (ERS) induced cell death and results in functional improvement after traumatic brain injury

Mark Nyanzu^1,2*, Felix Siaw-Debrah^1,2*, Pan Sishi^1,2, He Zibin Jin^1,2, Kunlin², Zhuge Qichuan^1,2, Huang Lijie^1,2

¹ Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China;

² Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China

*Co-authors contributed equally to this study.

Abstract:

Traumatic brain injury (TBI) is often caused by accidents that damage the brain. TBI can induce endoplasmic reticulum stress and lead to neuronal and glial cell death. Persistent overwhelming stimuli trigger ER stress to initiate apoptosis, autophagy, and cell death. Caspase-12 and CHOP signaling pathways are important players of ER stress, which is also modulated by ROS production, calcium disturbance, and inflammatory factors. In this study, we investigated the mechanism of cell death during the damage caused by TBI in vivo and in vitro, as well as the protective effect of resveratrol (RV), which is a polyphenol antioxidant found in red wine and has been shown to play a neuroprotective role. Here we report that Endoplasmic reticulum stress was activated in mice brains exposed to TBI. In the in vitro TBI model, apoptotic and neuroinflammatory cytokines were induced through the activation of BV2 microglial cell by LPS. LPS triggered activation of microglia, detected by an increase of TNF-α, and IL-1β, detected by qPCR. Treatment of RSV reduced neuroinflammation by inhibited LPS-induced TNF-α, IL-1β production in a concentration-dependent manner.

We investigated the effect of the administration of resveratrol (RSV) post-traumatic brain injury (TBI) on reducing ERS markers. TBI was induced by cortical contusion injury in mice. RSV (40 mg/kg in dimethyl sulfoxide) was administered intraperitonially at 5 min after TBI, followed by a daily dose for 3days. The expressions of GRP78, caspase-12, chop and caspase-3 were evaluated by Western blot. Neurological function was further evaluated to investigate the development of TBI. We found that post-TBI treatment with RSV could markedly inhibit the expressions of GRP78, CHOP and CASPASE-12. However, RSV treatment failed to reduce caspase-3 although neurological deficits at 72 h after TBI improved. These results indicated that RSV treatment could alleviate EBI after TBI, at least in part, via inhibition of ERS signaling pathway.

Keywords: Traumatic brain injury; Endoplasmic reticulum stress; Microglia, Resveratrol, Neuroinflammation

Modeling successful immune aging in long-lived mice

Abbe N. de Vallejo

Children's Hospital of Pittsburg, University of Pittsburgh, PA, USA

Abstract:

Human studies have led to identification of successful or exceptional aging, which is pertains to the maintenance of functional independence in the late years of life despite a long history of disease and/or pre-existing comorbid conditions. We have shown that successful agers have a unique repertoire of highly oligoclonal (pre)senescent NK-like T cells that are nevertheless functionally active capable of elaborating beneficial effector activities. To further understand the biology of successful aging, we have been studying mice that are deficient in PAPPA, pregnancy-associate plasma protein A, the enzyme that regulates the bioavailability of insulin-like growth factor (IGF) for signaling in tissues. PAPPA^-/- mice have ~40% extension in lifespan compared to their wildtype littermates. They are proportional dwarfs with normal growth curves and IGF/insulin levels and energy expenditure, as well as reduced incidence of spontaneous tumors and inflammatory pathology. Their most significant anatomical characteristic is their resistance to age-related degeneration of the thymus, the center to T cell selection and maturation. In this presentation, we will provide evidences for the maintenance of vigorous antigen-specific immunity in old animals in this long-lived strain. We will also show unpredicted evidences for vigorous innate function in old PAPPA^-/- mice in the setting of sepsis. And lastly, we will show unique brain structure associated with strong immunity in this successful aging model.

Keywords: Acquire immunity, Dendritic cells, Innate immunity, Longevity, Magnetic resonance imaging, Successful aging, T cells,

Comparison Between CT-guided Percutaneous Microwave Ablation and Thoracoscopic Lobectomy for StageⅠNon-Small Cell Lung Cancer

Bin LIU^1,2, Yongzheng WANG^1,2, Pikun CAO³, Wujie WANG^1,2, Wei WANG^1,2, Haiyang CHANG^1,2, Dong LI³, Xiao LI³, Xiaogang ZHAO⁴, Yuliang LI^1,2*

1.Department of Interventional Medicine, The Second Hospital of Shandong University, Jinan, Shandong Province, CN–250033, PR of China

2.Interventional Oncology Institute of Shandong University, Jinan, Shandong Province, CN–250033, China

3.School of Medicine, Shandong University, Jinan, Shandong Province, CN–250014, PR of China

4.Department of Thoracic Surgery, The Second Hospital of Shandong University, Jinan, Shandong Province, CN–250033, China

Abstract:

Objectives: To investigate the effectiveness and cost of CT-guided percutaneous microwave ablation (MWA) and thoracoscopic lobectomy for stageⅠ Non-Small Cell Lung Cancer (NSCLC) and to evaluate the efficacies of these two methods. Materials and Methods: 46 and 85 patients with stageⅠNSCLC treated with CT-guided percutaneous MWA or thoracoscopic lobectomy in our center from July 2013 to June 2015 were retrospectively analyzed, including the overall survival, disease free survival, local control rate, hospital stay and expense. Survival curves were constructed with the Kaplan-Meier method and compared by using the log-rank test. Result: The 1, 2 year-overall survival were estimated as 97.82%, 91.30% and 97.65%, 90.59% in MWA group and lobectomy group retrospectively. The 1, 2 year-disease free survival were 95.65%, 76.09% and 95.29%, 75.29%. Log-rank analysis of two sets of data was not significantly different (p=0.169). Hospital stay in MWA group and lobectomy group were 6.62±2.31 days and 9.57±3.19 days. Cost of MWA group and lobectomy group were 3274.50±233.91 and 4678.87±155.96 U.S. dollars. The differences were all significant (p=0.003). Conclusion: MWA and thoracoscopic lobectomy for stageⅠnon-small cell lung cancer demonstrate similar 1, 2 year-overall survival and disease-free survival, with no significant differences between the two groups, and shorter hospital stay & less expense. MWA should be considered the better choice for those patients with severe cardiopulmonary comorbidity, as well as those patients who were unwilling to accept surgery.

Key words: disease free survival; microwave ablation; non-small-cell lung cancer; overall survival; surgery

Supported by National Natural Science Foundation of China (61671276) and the Natural Science Foundation of Shandong Province (2014ZRE27479、ZR2018PH032).

Modified Percutaneous Transhepatic Papilla Balloon Dilation for High-Surgical-Risk Patients with Hepatolithiasis

Bin Liu^1,2, Pikun Cao³, Yongzheng Wang^1,2, Wujie Wang^1,2, Wei Wang^1,2, Haiyang Chang^1,2, Dong Li³, Xiao Li³, Yancu Hertzanu^1,4, Yuliang Li^1,2^＊

¹Department of Interventional Medicine, The Second Hospital of Shandong University, Jinan, Shandong Province, PR of China

²Interventional Oncology Institute of Shandong University, Jinan, Shandong Province, PR of China

³School of Medicine, Shandong University, Jinan, Shandong Province, PR of China

⁴Ben-Gurion University, Negev, Israel

Abstract:

Objectives: We aimed to evaluate the clinical efficacy of an innovative modified percutaneous transhepatic balloon dilation (PTBD) technique for removal of intrahepatic bile duct stones.

Methods: The data of 21 consecutive patients with intrahepatic bile duct stones, which underwent modified PTBD for clearance of gallbladder stones were retrospectively analyzed. With the aid of auxiliary apparatus, such as Fogarty catheter or basket, intrahepatic bile duct stones were pulled down into CBD, and pushed into duodenum with inflated balloon catheter. Outcomes recorded included operative time, postoperative hospital stay, success rate, causes of failure, and procedure-related complications. Biliary duct infection, hemorrhage, pancreatitis, gastrointestinal & biliary duct perforation, considered as short-term complications, were accessed before discharging. Long-term complications such as reflux cholangitis and calculi recurrence were monitored for 2 years.

Results: Intrahepatic bile duct stones were successfully removed in 20 (95.23%) patients. The median operative time was 65.8±5.3 (45-97) minutes. The median hospitalization duration was 10.7±1.5 (9-13) days. The mean hospitalization duration was 11.3 days. The procedure failed in one patient due to stone incarceration in intrahepatic bile duct. No pancreatitis or gastrointestinal & biliary duct perforation were confirmed in this research. During two year’s follow-up, there was no evidence of reflux cholangitis and calculi recurrence.

Conclusions: Modified PTBD is safe, feasible and effective treatment alternative for intrahepatic bile duct stones in a subgroup of patients with who cannot tolerate the risk of general anesthesia.

Key Words: intrahepatic bile duct; hepatolithiasis; Forgaty catheter; papilla

Sex-dependent aging in gait; results of the Korean Longitudinal Study on Cognitive Aging and Dementia (KLOSCAD) Gait Study

Hyang Jun Lee¹, Seonjeong Byun¹, Ji Won Han¹, Jun Sung Kim³, Euna Choi³, Ki Woong Kim^1,2,3,

1. Department of Neuropsychiatry, Seoul National University Bundang Hospital, Seongnam, Korea

2. Department of Psychiatry, Seoul National University, College of Medicine, Seoul, Korea

3. Department of Brain and Cognitive Science, Seoul National University College of Natural Sciences, Seoul, Korea

Objectives: We developed the normative data of the multi-points inertial measurement unit (IMU) sensor based gait analysis in healthy seniors, and investigated the impact of age and gender on gait parameters.

Methods: We enrolled 649 community-dwelling healthy elderly individuals aged 60 years or older (286 men and 363 women). We excluded the subjects who were diagnosed as frail based on the 5-item FRAIL scale. We also excluded the subjects who got less than 20 points in Tinetti Performance Oriented Mobility Test (POMA). We evaluated their gaits using 18 IMU sensors attached on their center of mass, forehead, neck, chest, both arms and legs. We asked each subject to stand up from a chair, walk 14m at their comfortable speeds, turn around, walk back and sit down for three times. We analyzed their gaits using the center 10m of steady walking period out of 14m walkway after discarding the 2m-acceleration and 2m-deceleration period at the ends. We compared gait characteristics between the demographic groups by age and sex. The IMU sensor data was extracted in time series data and processed with MATLAB 2016. Statistics were performed by SPSS 18.00.

Results: Compared to women, men showed the larger arm movement (t= 9.475, p<0.000) and smaller pelvic yaw angle (t= -8.836, p<0.000) and torso yaw angle (t= -8.782, p<0.000). With advancing age, gait speed (b = -0.292, p < 0.000) and maximum accelerations at feet (b = -0.214, p < 0.000) and legs (b = -0.129, p = 0.001) decreased while the acceleration of arms (b = -0.077, p = 0.051) and the angular difference of arms (b = 0.095, p = 0.016) and legs (b= -0.067, p = 0.088) modestly changed.

Interpretation or conclusion: With advancing age, elderly gait changed. In speed and foot accelerations which were directly connected gait performance, subjects showed distinctive decrease. The arm movement in angle and acceleration showed that the subjects had slower movement but didn’t show visible change in operating range. The elderly gait had characteristics of slowness but maintained visible form.

Differentiated embryo chondrocyte 1 (DEC1) is a novel negative regulator of hepatic fibroblast growth factor 21 (FGF21) in aging mice

Ujjal K. Bhawal¹ and Makoto Makishima²

¹Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, Chiba 271-8587, Japan.

²Division of Biochemistry, Department of Biomedical Sciences, Nihon University School of Medicine, Tokyo 173-8610, Japan

Abstract:

Human differentiated embryo chondrocyte expressed gene 1 (DEC1) is frequently used as a marker of senescence in vivo. Fibroblast growth factor 21 (FGF21), a novel endocrine-like member of the FGF superfamily, is highly expressed in the liver, and FGF21-transgenic mice have extended lifespans. Thus, we hypothesized that FGF21 may play a role in the DEC1-mediated aging process. In this study, DEC1 knockout (KO) mice were used to characterize the mechanism by which FGF21 protects mice from aging. Aging is strongly diminished in DEC1 KO mice, which is reflected by decreased lipid levels and oxidative stress, leading to an amelioration of liver function and structure. The expression of FGF21 decreased with aging in wild-type (WT) mice, whereas ATF4, Phospho-ERK and Phospho-p38 expression was maintained and was accompanied by a compensatory rise of FGF21 mRNA and protein expression in DEC1 KO mice. Over-expression of DEC1 markedly abolished the hepatic expression of FGF21, and siRNA-mediated inhibition of endogenous DEC1 increased the expression of FGF21. DEC1 further diminished the expression of ATF4 in HepG2 cells over-expressing DEC1. The induction of FGF21 and ATF4 at the mRNA and protein levels during the course of aging supports the view that DEC1 KO mice are able to restore the age-related imbalance of metabolism. Collectively, the data obtained in this study suggest that DEC1 is a novel negative regulator of hepatic FGF21 expression.

Mutant erythropoietin enhances white matter repair via JAK2/STAT3 pathway in middle aged mice after cerebral ischemia

Rongliang Wang^1,2,3, Zhibin Fan^1,2,3, Fei Ju⁴, Zhen Tao^1,2,3, Haiping Zhao^1,2,3*, Yumin Luo^1,2,3*

¹Cerebrovascular Diseases Research Institute and Department of Neurology, Xuanwu Hospital of Capital Medical University, Beijing 100053, China

²Center of Stroke, Beijing Institute for Brain Disorders, Beijing 100053, China

³Beijing Key Laboratory of Translational Medicine for Cerebrovascular Diseases, Beijing 100053, China

⁴Internal Medicine Department, Central Hospital of Beijing Prison Administration Bureau, Beijing 100054, China

Abstract:

Previous studies indicated that EPO maintained the M2 microglia phenotype that contributes to white matter repair after ischemic stroke in young mice (2-month old). However, high doses and multiple administrations of EPO may cause deleterious side effects due to its erythropoietic activity. This study investigates the neuroprotective effects of non-erythropoietic mutant EPO (MEPO) on white matter and underlying mechanism in middle aged (8-month old) mice following cerebral ischemia. Methods: A total of forty 8-month old male C57 BL/6 mice were treated with MEPO (5000 IU/kg), or vehicle after focal middle cerebral artery occlusion (MCAO). In order to confirm the molecular mechanisms involved in the effects of MEPO on regulating microglial polarization. A specifically inhibitor, AG490 was used to block the JAK2/STAT3 pathway. Neurological function was assessed by beam walking and adhesive remove test. The body weight and brain tissue loss of each experimental group were calculated. Immunofluorescence staining, and western blotting were used to assess the severity of white matter injury and the phenotypic changes of microglia. Bromodeoxyuridine (BrdU) was injected intraperitoneally daily for measurements of newly proliferating oligodendrocyte. Results: MEPO significantly improved neurobehavioral outcomes, alleviated brain tissue loss, ameliorated white matter injury after MCAO compared with the vehicle group (p<0.05). Cerebral ischemia promoted microglial polarization toward the M1 phenotype, whereas MEPO promoted oligodendrocytogenesis by shifting microglia toward M2 polarization 14 days after cerebral ischemia-reperfusion (p<0.05). However, the MEPO’s effect on microglial M2 polarization and oligodendrocytogenesis was largely suppressed by AG490 treatment (p<0.05). Conclusion: MEPO treatment improves white matter integrity after cerebral ischemia, may be partly explained by that MEPO facilitates microglial polarization toward the beneficial M2 phenotype to promote oligodendrogenesis, which possibly mediated by JAK2/STAT3 signaling. This study provided a novel insight into MEPO treatment for ischemic stroke.

Keywords: Cerebral ischemia, Mutant erythropoietin, JAK2/STAT3 pathway, White matter repair, Oligodendrogenesis

Interaction of psychostimulant abuse, aging and disease

Michael Foster

Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, Fort Worth, TX 76107, USA

The latest reports published by agencies such as Substance Abuse and Mental Health Services Administration and the National Survey of Drug Use and Health have indicated a steady rise of substance use between the ages of 50-64. However, the potential health care issues stemming from increased drug use in the aging population is poorly understood and is unlikely to be resolved through clinical study due to the complexity of patient medical and drug-taking history. To better understand the nature of the aging/substance abuse interaction, we have studied the effects of psychostimulants and other abused drugs under the following conditions: (i) In drug naïve mice from separate age groups representing young adulthood, middle age, and senescence; (ii) In adults exposed to psychostimulants during adulthood and maintained drug- free into advanced age. Our results suggest that responsiveness to psychostimulants and psychostimulant-induced plasticity are markedly different in older drug-naïve age groups when compared with young adults. In other studies, young adult mice exposed to psychostimulants at doses much lower than those modeling chronic abuse, were profoundly impaired in tests of psychomotor and cognitive functions when tested at advanced age. These results suggest that drug abuse history may profoundly influence health, susceptibility to addiction and treatment outcomes in persons of advanced age.

Accelerated aging in Zmpste24 deficient mice result in attenuated experimental lung fibrosis.

Annie Pardo Jazmín Calyeca, Moisés Selman. Facultad de Ciencias,

Universidad Nacional Autónoma de México, Instituto Nacional de Enfermedades Respiratorias Ismael Cosío Villegas.

Abstract:

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and lethal disease of unknown etiology. Its incidence and prevalence increase with age, however, the mechanisms linking IPF to aging are uncertain. Experimental models in “normal” aging mice are scanty mainly because these animals are time-consuming and expensive. Recently, it was demonstrated that deletion of the gene Zmpste24 in mice causes an accelerated aging phenotype (including scoliosis, muscular dystrophy and dilated cardiomyopathy). Thus, we aimed to evaluate the lung response to a fibrotic injury in this accelerated aging mouse. Young (4 weeks) and old (16 weeks) Zmpste24 deficient mice and young (4 weeks) and old (82 weeks) WT littermate were examined for the fibrotic response at 21 days after bleomycin-induced lung injury. Young animals showed no differences in the extent and severity of lung fibrosis as analyzed by morphology and hydroxyproline quantification. By contrast, aged Zmpste24 deficient mice showed significant less fibrosis compared with aged wildtypes counterparts. The attenuation of the fibrotic response was confirmed by a significant decrease in lung collagen content. Global gene expression analysis revealed an increased expression of several antifibrotic microRNAs including miR23a, miR27a, miR29a and miR29b-1 in bleomycin damaged lungs of Zmpste24 mice, which was validated by qPCR. As expected, several targets of these microRNAs, including some extracellular matrix proteins were decreased. Our results suggest that the absence of Zmpste24 protects aged mice from the development of bleomycin-induced pulmonary fibrosis at least partially through the over-expression of microRNAs with antifibrotic activity. Funding: This research was supported by CONACYT 281074

Title of the talk: Na/H exchanger regulates glioma tumor microenvironment and immunogenicity

Dr. Dandan Sun, MD, PhD,

Department of Neurology, University of Pittsburgh, USA

Abstract:

The prognosis of malignant glioma patients remains poor under the standard therapies and the weak immunogenicity of gliomas presents a barrier for effective immunotherapy. Na/H exchanger isoform 1 (NHE1) maintains alkaline intracellular pH (pH_i) of glioma cells and immunosuppressive acidic extracellular pH (pH_e). In addition, NHE1 is expressed in tumor-associated microglia and macrophages (TAMs) and involved in communications between glioma and TAMs. Therefore, we speculate that NHE1 plays a role in developing tumor resistance to temozolomide (TMZ)-mediated chemotherapy and immunosuppressive tumor microenvironment. In this study, we investigated whether pharmacological inhibition of NHE1 presents a novel strategy for combinatorial therapies. Here, we show that TMZ treatment increased NHE1 protein expression in two syngeneic mouse glioma models. Pharmacological inhibition of NHE1 potentiated the cytotoxic effects of TMZ, leading to inhibition of tumor growth, abrogated tumor invasiveness, and increased median survival of mice. Especially, blockade of NHE1 stimulated proinflammatory activation of TAM and increased cytotoxic T cell infiltration into tumors. Combining TMZ, anti-PD-1 antibody treatment with NHE1 blockade prolonged the median survival in the mouse glioma models. These results clearly demonstrate that NHE1 protein plays a role in glioma survival and migration. Pharmacological blockade of NHE1 activity enhanced TMZ-induced cytotoxicity, and increased tumor immunogenicity to improve glioma immunotherapy.

Keywords: Anti-PD-1 antibody therapy, glioma, immune response, Na/H exchanger isoform 1 inhibitor HOE642, temozolomide, tumor microenvironment

How to favor clinical trials for longevity: A Manhattan project against senescence?

Didier Coeurnelle

Heales (Healthy Life Extension Society, Brussels, Belgium)

Abstract:

Diseases related to old age are responsible for 70 % of deaths worldwide and 90 % in countries like France and the USA. The only way to prevent illnesses linked to aging is to attack the root cause – that is aging itself. It is time to start working towards solutions to this universal human tragedy. Aging doesn’t just cause innumerable deaths, it is also the source of many debilitating illnesses like Alzheimer’s disease, cardiovascular diseases, muscle wasting, decline of vision and hearing, osteoporosis, rheumatoid arthritis. A longer and healthier life is enjoyed by the citizens who can benefit from it. Longevity is potentially useful for a sustainable environment and for a peaceful society.

How to inform ethicists, researchers and stakeholders? How can we have a "Moonshot longevity vision"? Can we consider research for a longer life as a moral obligation, a duty to rescue, a guarantee for a better future world? It is necessary to compare common aspects of the ideas of the mainstream stakeholders and the general public to the objectives of "longevists" concerning: Environmental aspects, Questions related to non-violence, Objectives of equality and equity, Priority for the weakest/oldest/frailest.
We should favor the possibility for well-informed citizens to make clinical tests for longevity in the following fields: Genomics and gene therapies, new drugs, stem cells, restorative nanotechnologies. There are improvements possible for the testing protocols, the recruitment procedures, the 11th Revision of the International Classification of Diseases and the legal frameworks, especially the rules making it more difficult for older patients to take part in clinical tests. This will enhance the existing testing capabilities of many involved scientists.
Heales (Healthy Life Extension Society) and the ILA (International Longevity Alliance) are among the organizations who are working resolutely in this direction. The current state of policy and funding for aging research, especially in France and other countries of the EU is promising.

Key words: Longevity advocacy, clinical trials, European funding

The pipeline to develop solutions to biological aging

Anton Kulaga

Computational Biology of Aging Group, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania; International Longevity Alliance

Miriam Leis

International Society on Aging and Disease (ISOAD), Munich, Germany

Abstract:

Although aging-related diseases (ARD) are being recognized as a great challenge around the world, with dementia alone expected to affect 152 million people globally by 2050, the development of dedicated medical treatments is stalling. Pharmaceutical companies have been forced to stop related clinical trials and recall their drugs due to failure. This caused an overall decline in research and investments by pharmaceutical companies, especially in regard to Alzheimer’s or Parkinson’s disease.

In view of the tremendous social and economic problems societies will face due to the increase in ARDs, it is necessary to explore alternative approaches in research and development. Instead of the traditional focus on the treatment of single diseases out of several dozen ARDs, the relatively young research field of biology of ageing (BA or biogerontology) is focussing on the scientific understanding of the underlying ageing process itself that is deemed to be the root cause of a multitude of ARDs. Although this holistic research approach of BA has uncovered new, promising hypothesis about the relations between different ARDs, it is a complex endeavour that requires international collaborative efforts. Describing the BA technology landscape in terms of available research methods and technologies, theories and translational pipelines on different technology readiness levels, forms the basis for identifying bottlenecks, extending the scientific and technological state-of-the-art and choosing promising paths for future R&D.

Keywords: aging-related diseases, biogerontology, translational pipeline, technology readiness levels

Global Aging: UN in Action

Danan Gu

(United Nations Population Division)

Abstract:

The world’s population has been aging rapidly over the last few decades and the pace will continue to increase into the foreseeable future. Population aging is one of the greatest social transformations in the twenty-first century. Although the increase in the number of older adults represents a triumph of human development and civilization, it also poses tremendous challenges to all sectors of every society—particularly for achieving sustainable development goals, ensuring healthy lives, and promoting well-being at all ages. It is more important than ever that governments design innovative policies and public services specifically targeted to older persons, including those that address housing, social security, healthcare, infrastructure, and other social protections. The United Nations (UN) has been a pioneer on this forefront and the shepherd in promoting healthy aging by addressing global aging issues over the last several decades. Providing global leadership, the UN has played a critical role in formulating resolutions, guidelines, program actions, and frameworks for addressing the many challenges of global aging. Within the headquarters and regional offices of the UN system, new offices and platforms have been established and the routine mandates of many existing UN entities have been incorporated (or shifted) to adapt to such global-social transformations. In working with member states, other international organizations, intergovernmental organizations, NGOs and civil societies, the UN has made substantial efforts —through multiple channels— to initiate, advocate, and organize interactive discussions, annual commissions, high-level dialogues, panels, platforms, and forums to address challenges of global aging, covering various thematic issues from the protection of human rights and the dignity to long-term and palliative care; from retirement life to poverty reduction; and from quality of life to living environment, and from data collection to program evaluation. The UN is now working with member states, NGOs, and civil societies to monitor the system of sustainable development goals. This presentation will review the efforts, contributions, and commitments of the UN in addressing the challenges of global aging.

Keywords: Global Aging, United Nations, Healthy Ageing, Sustainable Development Goals,

Public advocacy as a crucial part of longevity R&D and therapy

Ilia Stambler

Israeli Longevity Alliance; Vetek (Seniority) Association – the Senior Citizens Movement (Israel); International Society on Aging and Disease (ISOAD)

Abstract:

Public support is a necessary condition for enhancing the research, development and application of emerging longevity therapies. Public advocacy is a primary instrument for increasing the support, hence longevity advocacy is an indispensable part of the longevity field. Several recent longevity advocacy campaigns in Israel and internationally, demonstrate the utility of advocacy for promoting public engagement, research and health policy in favor of aging and longevity R&D. More support for and involvement in longevity advocacy is needed by the scientific community and the broad public.

Key words: longevity advocacy, public support, longevity R&D and therapy

Long-term Survival and Regeneration of Neuronal and Vasculature Cells in the Core Region after Ischemic Stroke in Mice

Shan Ping Yu

Department of Anesthesiology, Emory University School of Medicine, Atlanta, GA 30322; Center for Visual and Neurocognitive Rehabilitation, Atlanta Veterans Affair Medical Center, Decatur, GA, 30033， USA

Abstract:

Focal cerebral ischemia results in an ischemic core surrounded by a peri-infarct region (penumbra). It is widely believed that neurons in the ischemic core die in a few days after stroke while cells in the penumbra can survive much longer and has been regarded as the primary therapeutic target. In the present investigation, we tested the hypothesis that, inside the ischemic core, some neuronal and vascular cells could survive the initial ischemic insult and regenerative niches could exist many days after stroke. Adult male mice were subjected to focal ischemic stroke. This ischemic insult uniformly reduced the local cerebral blood flow (LCBF) by 90%. Massive cell death and a significant infarction were cultivated in the ischemic cortex 1-3 days later. Nevertheless, significant levels of trophic/growth factors BDNF and VEGF remained in the core tissue, some NeuN-positive and Glut-1/College IV-positive cells with intact ultrastructural features still resided inside the core 7-14 days post stroke. BrdU-positive and TUNEL-negative neuronal and endothelial cells existed in the core at the same time, while extensive extracellular matrix structural networks developed in the region. Meanwhile, GFAP-positive astrocytes accumulated in the penumbra while Iba-1-positive microglial/macrophages invaded the ischemic core several days after stroke. These data suggest that the ischemic core remains an actively regulated brain region with residual but viable neuronal and vascular cells acutely and chronically after stroke. The remaining and regenerating cells and neurovascular structures may provide a minimal but indispensable infrastructure for tissue repair in the post-stroke brain.

Unraveling the mechanisms that master epithelial regeneration in the adult kidney

Margo Montandon, Lucile Yart, Catherine Pons, Lou Duret, Valérie Vial, Magali Mondin, Eric Gilson and Marina Shkreli

Institute for Research on Cancer and aging, Nice (IRCAN), Nice 06107, France

UCA, CNRS UMR7284, INSERM U1081

Abstract:

In contrast to the invertebrates (drosophila) and the basally branching vertebrates (zebrafish), mammals are thought to be unable to generate new nephrons under physiological or pathological conditions. Nonetheless, efficient regeneration of the cells shaping the mammalian kidney tubules is observed following injury. However, the identity of the stem cells that support kidney tubules replenishment remains unclear. In contrast to tubular epithelia, the highly specialized glomerular epithelial cells named podocytes show limited renewal capacities in the adult mammals, and unavailing podocyte regeneration in response to injury ultimately leads to the development of end-stage renal failure. Remarkably, efficient and fast podocyte regeneration has been described in a telomerase-induced regeneration mouse model. Here we show that renewed podocytes observed in this telomerase-enforced regeneration system are generated by activation of renal stem cells. We found using an unbiased stochastic multicolor tracing approach that efficient podocyte regeneration within the adult mammalian kidney is achieved by recruitment of an extra-glomerular stem cell population. This finding shows that ectopic transient activation of telomerase within the adult kidney is an efficient way to activate a latent podocyte regenerative program that involves resident stem cells and support the notion that telomerase carries potent effects on adult stem cells in mammals. Modulating telomerase therefore appears to be an appealing approach for regenerative medicine and for handling degenerative diseases in humans.

New methods for the diagnosis of old-age multimorbidity

Ilia Stambler¹, David Blokh², Eliyahu H. Mizrahi³

¹Department of Science, Technology and Society, Bar Ilan University, Ramat Gan, Israel

²C.D. Technologies Ltd., Beer Sheba, Israel

³Geriatric Medicine and Rehabilitation Department B, The Geriatric and Medicine Center "Shmuel-Harofe”, Beer Yaakov, Affiliated to Sackler School of Medicine, Tel-Aviv University, Tel-Aviv, Israel

Abstract:

Elderly patients are commonly characterized by the presence of several chronic aging-related diseases at once, or old-age “multimorbidity,” with critical implications for diagnosis and therapy. However, at the present, there is no agreed or formal method to diagnose or even define “multimorbidity.” There is also no formal quantitative method to evaluate the effects of individual or combined diagnostic parameters and therapeutic interventions on multimorbidity. We outline two information-theory based methodologies to provide such a measurement and definition, using the information theoretical measure of normalized mutual information. A cohort of geriatric patients, suffering from several age-related diseases (multimorbidity), including Ischemic Heart Disease, COPD, Dementia, and Hip Fracture, were evaluated by a variety of diagnostic parameters, including static as well as dynamic biochemical, functional-behavioral, immunological and hematological parameters. In the first method, multimorbidity was formally coded and measured as a composite variable of several chronic age-related diseases. In the second method, we evaluated the informative values of particular parameters or parameter combinations on particular diseases separately, and then combined the ranks of informative values to provide an overall estimation about several diseases at once. The normalized mutual information allowed establishing the exact informative value of particular parameters and their combinations about the multimorbidity values. With the currently intensifying attempts to reduce aging-related multimorbidity by therapeutic interventions into their underlying aging processes, with the addition of more clinical data, the proposed methodologies may outline a valuable direction toward the formal indication and evidence-based evaluation of effectiveness of such interventions. In addition to developing the methodologies to evaluate old-age multimorbidity, the normalized mutual information was also used to create diagnostic decision rules to predict hip fractures in order to facilitate their prevention.

DNA damage responses in aging and disease

Björn Schumacher

Institute for Genome Stability in Aging and Disease, Medical Faculty, Cologne Excellence Cluster for Cellular Stress Responses in Aging-Associated Diseases (CECAD) Research Centre and Centre for Molecular Medicine (CMMC), University of Cologne, Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany

ABSTRACT:

The causal contribution of DNA damage in driving the aging process has become evident in a variety of progeroid syndromes that are caused by defects in DNA repair systems. Congenital defects in genome maintenance mechanisms cause complex disease phenotypes characterized by developmental growth failure, cancer susceptibility, and premature aging. The distinct human disease outcomes of DNA repair defects are particularly apparent in syndromes caused by mutations in nucleotide excision repair (NER). While transcription-coupled (TC-) NER defects lead to growth and mental retardation and premature ageing in Cockayne syndrome (CS) patients, global-genome (GG-) NER mutations lead to highly skin cancer prone Xeroderma pigmentosum (XP). Intriguingly, the distinct outcomes of NER deficiencies are conserved in the simple metazoan C. elegans. TC-NER deficiency renders worms highly susceptible to DNA damage during developmental growth and with aging, while GG-NER defects give rise to genome instability in proliferating germ cells.

We employed the nematode model to investigate distinct DNA damage response (DDR) mechanisms in (postmitotic) somatic tissues and in the germline. DNA damage that persists in somatic tissues leads to activation of the insulin-like growth factor signalling (IIS) effector DAF-16. The FoxO transcription factor DAF-16 is efficiently activated in response to DNA damage during development while its DNA damage responsiveness declines with aging. We demonstrated that DAF-16 alleviates growth arrest and enhances DNA damage resistance in somatic tissues even in the absence of DNA repair. We propose that IIS mediates DNA damage responses in somatic tissues and that DAF-16 activity enables developmental growth amid persistent DNA lesions and promotes maintenance of differentiated tissues through enhanced tolerance of DNA damage that accumulates with aging. Mechanistically, we determined that the conserved ERK1/2 MAPK pathway regulates the DAF-16-mediated DDR.

An integrated proteomics, phospoproteomics and lipidomics analysis of the in vivo response to persistent UV-induced DNA lesions revealed a comprehensive picture of the organism’s DDR. We determined a shift in proteostasis towards autophagy, a dampening of glucose and lipid metabolism, and functionally implicate IIS, EGF-, and AMPK-like signalling. We found striking similarities between the acute response to DNA damage and the proteome of aging animals thus further supporting the underlying role of DNA damage accumulation in the aging process. Our data provide new insights into the organism’s response program to DNA damage during development and aging and suggest new intervention targets for triggering stress responses to antagonize the detrimental consequences of genome instability.

Keywords: Aging, DNA damage, nucleotide excision repair, progeroid syndromes, C. elegans

Some methodological characteristics of Alzheimer-associated urine neuronal thread protein detected by enzyme-linked immunosorbent assay

He Jin^a, Rong Wang^a,b*, Zhaohui Liu^c, Qiang Jia^c, Yanchuan Wu^a, Zhiwei Zhao^a, Yulan Wang^a, Xu Zhang^a

^aCentral Laboratory, Xuan Wu Hospital, Capital Medical University, Beijing Geriatric Medical Research Center, Key Laboratory for Neurodegenerative Disease of Ministry of Education, Beijing, China

^bBeijing Institute for Brain Disorders, Beijing, China

^Department of nephropathy, Xuanwu Hospital, Capital Medical University, Beijing, China.

ABSTRACT:

Background: Alzheimer-associated neuronal thread protein (AD7c-NTP) has been found to be a candidate biomarker of Alzheimer’s disease (AD).

Objective: To investigate the effects of urine collected time, different preservatives addition and storage condition on the measurement of urine AD7c-NTP by enzyme-linked immunosorbent assay (ELISA).

Methods: Three hundred urine samples were collected from 20 participants at three-time points on five consecutive days. These samples were immediately placed at 4°C and detected within 2h. The single spot samples of the first day morning was split into eleven duplicate aliquots (a-k) of 1 ml each, (a) without any preservative (untreated), (b) containing boric acid (2 g/L), (c) containing NaHCO₃ (5 g/L), (a-c) were detected at six different time points. For the other eight preservative-free samples, (d)-(g) were stored at −20°, (h)-(k) were stored at −70°C, respectively, detected at different time points. All of the results were compared with the baseline urine.

Results: The urine AD7c-NTP levels at different time points behaved stably (P>0.05). Urine samples without any preservative increased over time, and compared with the NaHCO₃ addition group, boric acid addition group behaved stably. Samples stored at −20°C and −70°C led to an obviously false positive.

Conclusions: AD7c-NTP can be tested using random urine instead of the first morning urine. If the specimen cannot be tested in time, boric acid appears to be an acceptable preservative with storage at 4°C, freezing is not recommended.

Keywords: Alzheimer’s disease, urine, biomarker

COL19A1 levels improve prognosis in Amyotrophic Lateral Sclerosis

Ana C. Calvo^1*, Gabriela Atencia Cibreiro², Paz Torre Merino², Juan F. Roy³, Adrián Galiana⁴, Alexandra Juárez Rufián², Juan M. Cano⁵, Miguel A. Martín², Laura Moreno¹, Pilar Larrodé¹, Pilar Cordero Vázquez², Lucía Galán⁶, Jesús Mora⁷, José L. Muñoz-Blanco⁸, María J. Muñoz¹, Pilar Zaragoza¹, Elena Pegoraro⁹, Gianni Sorarù⁹, Marina Mora¹⁰, Christian Lunetta¹¹, Silvana Penco¹², Claudia Tarlarini¹², Jesús Esteban², Rosario Osta¹, Alberto García Redondo²

¹Laboratory of Genetics and Biochemistry, Faculty of Veterinary-IIS, IA2-CITA, University of Zaragoza, Spain

²NEUROLOGY Department, ALS UNIT. CIBERER U-723. Health Research Institute, 28041 Madrid, Spain

³Ferkauf Graduate School of Psychology, Yeshiva University, Bronx, NY 10461, USA

⁴Servicio de Reumatología, Hospital Universitario de la Princesa, Instituto de Investigación Sanitaria La Princesa, 28006 Madrid, Spain.

⁵Orthopaedic Surgery Department, October 12^th Hospital, 28041 Madrid, Spain.

⁶Neurology Department, ALS Unit. Clínico Universitario San Carlos Hospital, Profesor Martín Lagos, s/n, 28040 Madrid, Spain.

⁷Neurology Department, ALS Unit. Carlos III Hospital, 28029 Madrid, Spain.

⁸Neurology Department, ALS Unit. HEALTH RESEARCH INSTITUTE, Gregorio Marañón Hospital “IISGM”. 28007 Madrid, Spain.

⁹Neurological Clinic, Department of Neurosciences, University of Padova, 35122 Padova, Italy.

¹⁰Muscle Cell Biology Laboratory, Neuromuscular Diseases and Neuroimmunology Unit, Fondazione IRCCS Istituto Neurologico C. Besta, 20133 Milan, Italy.

¹¹NEMO (NEuroMuscular Omnicentre) Clinical Center. Fondazione Serena Onlus. Piazza Ospedale Maggiore 3, 20162 Milan, Italy.

¹²Medical Genetics Unit, Department of Laboratory Medicine, Niguarda Ca’ Granda Hospital. P.za Ospedale Maggiore 3, 20100 Milan, Italy

Abstract:

Background: Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease of unknown origin that prompts a progressive muscle weakening and a motor neuron loss. The identification of more reliable diagnostic or prognostic biomarkers in ALS is urgently needed. The objective in this study was to look at the associations among molecular markers and clinical variables to identify more reliable prognostic biomarkers mirroring neurodegeneration that could be of help in clinical trials.

Methods and Findings: This study included a total of 268 participants from three cohorts, matched for age and gender, whenever possible: Muscle biopsy cohort (87 participants), Blood cohort (141 participants) and Serial blood cohort (40 participants). The muscle and blood cohorts were analyzed in two cross-sectional studies. The serial blood cohort was analyzed in a longitudinal study at 6-monthly intervals. The expression levels of fifteen target genes and fourteen proteins involved in muscle physiology and differentiation, as well as metabolic processes and neuromuscular junction dismantlement were studied by real time PCR and Western Blot, respectively, in each sample from the three cohorts.

In the muscle biopsy cohort, the risk for a higher mortality in an ALS patient that showed high COL19A1 protein levels and a fast progression of the disease was 70.5% (ALSFRS-R slope, hazard ratio, HR: 1.569, CI: 1.245-1.976, p=0.000; COL19A1 protein levels, HR: 2.158, CI: 1.020-4.566, p=0.044). In the blood cohort, the risk for a higher mortality in an ALS patient that showed high COL19A1 gene levels and a fast progression of the disease was 92.47% (ALSFRS-R slope, hazard ratio, HR: 5.798, CI: 3.405-9.872, p=0.000; COL19A1 levels, HR: 2.291, CI: 1.435-5.945, p=0.003). In relation to the Serial blood cohort, the linear mixed model analysis showed a significant relationship between increasing COL19A1 levels along disease progression and a faster progression during the follow-up period of 24 months in each patient (Estimate: -0,0093, CI: (-0,0181) - (-0,00039); p=0,042). Time-dependent Cox regression analysis suggested that the risk for a higher mortality in a sporadic ALS patient that showed higher COL19A1 levels and a faster progression was 17.9% (95% CI 4.6%-32.7%, P=0.007). This study was conducted in seven centres from Spain, France and Italy from January 2010 to February 2016. Our main limitation of this study includes that other genetic cases of ALS different from SOD1 and C9ORF72 mutants could not be enrolled and therefore, the role of COL19A1 in these genetic cases should be also explored.

Conclusions: We provide evidence that the association of COL19A1 expression levels and the disease progression in muscle biopsies and blood from ALS patients can support an earlier prognosis of the disease, especially in those patients that share a high disability in the first symptomatic stages of the disease. The accurate monitoring of ALS patients at clinical and molecular levels is relevant to follow the disease progression. Consequently, COL19A1 could be considered a candidate biomarker that could help the selection of a homogeneous group of patients for upcoming clinical trial and may be pointed out as a promising and a novel therapeutic target in ALS.

GUANOSINE PROMOTES PROLIFERATION AND NEUROGENESIS IN ADULT MICE BUT NOT AFTER CHRONIC TREATMENT IN AGING MICE

Tetsade Piermartiri1, Beatriz Santos2, Fernanda Barros3, Rui D Prediger3, Carla I Tasca2

Universidade Federal de Santa Catarina - UFSC, 1 Programa de Pós-Graduação em Neurociências, 2 Departamento de Bioquímica, 3 Departamento de Farmacologia, Florianópolis, Brazil

Abstract:

In the mice adult brain, neural stem cells (NSCs) are capable of differentiating generating new neurons in a process called neurogenesis, in the subventricular zone (SVZ) and in the dentate gyrus (DG) of hippocampus. The guanine nucleoside, guanosine (GUO), may act as neuromodulatory agent and has a protective role on neurodegenerative processes. We are now investigating whether in vitro GUO treatment (100 μM, for 7 days) affect NSCs in DG of adult mice. GUO promotes proliferation (P<0.05) of NSC forming neurospheres and NSCs differentiation (P<0.05). We wonder where GUO would have similar effects in vivo. The adult mice (C57BL/6) were treated with GUO (8 mg/kg, i.p, intraperitoneal, for 26 days) and 5-bromo-2'-deoxyuridine (BrdU 50 mg/kg, i.p, for 5 days). GUO increases (P<0.05) the number of BrdU+ cells, and neuronal differentiation by the percentage of BrdU+, doublecourtin (DCX)+ cells colocalization in DG. No difference was found in SVZ. Behavior assay showed that GUO does not change general heath or cognitive tests, however exert an antidepressant-like behavior in the tail suspension test (P<0.05). In mice, adult neurogenesis drastically diminishes with aging, and sharply drops around 7 months. To circumvent age-related reductions in neurogenesis, we design an experimental treatment starting at 7 months until 9 months old with GUO (8 mg/kg, i.p, daily), and BrdU (50 mg/kg, i.p for 5 days). However, GUO fail to increase neurogenesis in aging mice, and did not change behavior. Our findings suggest that the mechanism of purinergic signaling on NSCs and neurogenesis occurs primarily in adult mice.

Hypoxia post-conditioning induced neuroprotection against cerebral ischemic injuries in mice via promoting glycolysis

Changhong Ren^{1, 2,} Rongrong han¹, Haiyan Li^{1, 2}, Sijie li¹, Yuanyuan Liu¹, Xunming Ji^{1, 2#}

1. Beijing Key Laboratory of Hypoxia Conditioning Translational Medicine

2. Institute of Hypoxia Medicine, Xuanwu Hospital, Capital Medical University

Abstract

Ischemic stroke initiated by transient or permanent cerebral blood flow decline remains the leading cause of permanent disability in industrialized nations. However, therapeutic strategies to improve patient recovery are still limited. Hypoxia post-conditioning (HPostC) has been known to be neuroprotective against ischemic injuries in vivo and in vitro. Understanding its mechanism of action may promote its clinical translation. In the present study, we devised our study to test a HPostC protocol on protection of a focal cerebral ischemic induced injury and to explore the underling mechanism. We found that our HPostC method improved energy supply by elevating the level of glucose，pyruvate and ATP/ADP ratio in cerebral hemisphere in mice. In distal middle cerebral artery occlusion (dMCAO) in mice, this HPostC protocol reduced infarct size, and associated with increased levels of pyruvate, pyruvate/lactate ratio and ATP/ADP ratio in the dMCAO mice. Western blot analysis indicated that the HPostC protocol up-regulated AMPK signaling activities in cerebral hemisphere. Our results suggest that this HPostC protocol exerts its neuroprotective effect by promoting glycolysis to elevate the ATP/ADP level, and the AMPK/KFPFB3 signaling pathway. This result may provide biomarkers for clinical use of HPostC methods.

Key words: hypoxic conditioning, stroke, metabolism

The effect of hypoxia preconditioning on DNA methyltransferase and PP1γ in hippocampus of hypoxia preconditioned mice

Guo shao

Inner Mongolia Key laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, 104060, Inner Mongolia Autonomous Region, China

Abstract:

It is well known that hypoxia preconditioning can increase hypoxic tolerance by changing the expressions of some genes in the brain. DNA methylation is important for regulating gene expression and is catalyzed by DNA methyltransferase (DNMT), an enzyme that is abundant in the brain. The impact of hypoxia preconditioning on DNA methylation remains unknown. Mice were randomly divided into three groups: blank control group with no exposure to hypoxia (H0), the hypoxia control group exposed to hypoxia once (H1), and the hypoxia preconditioning group exposed to 4 runs of hypoxia (H4). The mRNA and protein level of three kinds of DNMTs and the activity of DMNTs together were detected. Protein phosphatase 1(PP1)γ, which critically regulates neuroprotective pathways in brain, was measured in mRNA, protein, activity and promoter methylation. DNMT1 was unchanged in H1 and H4, DNMT3A and DNMT3B were decreased in H4. The mRNA and protein levels of PP1γ were decreased in H4. However, there was no detectable change in the level of DNA methylation of the promoter of PP1γ (-321 bp to 95bp). These findings suggest that DNA methylation may have a role in hypoxia neuroprotection, and the change of PP1γ, which did not depend on the change of its promoter (-321 bp to 95bp) DNA methylation, may be involved in neuroprotection.

The Activation of Full-Length Tropomyosin-Related Kinase Type B Receptors Signaling Regulated by Rno-miR-185-3p Suppresses the Epileptiform Discharges in Cultured Hippocampal Neurons

Zhanjun Yang

Baotou Medical College, Baotou, 104060, Inner Mongolia Autonomous Region, China.

Abstract:

Epilepsy is a common neurological disorder characterised by occurrence of spontaneous recurrent epileptiform discharges (SREDs) in neurons. The cellular mechanisms that underlie epilepsy are known to be regulated by brain-derived neurotrophic factor. However, some studies show that tropomyosin-related kinase B (TrkB), which is the receptor for BDNF, plays an important role in the pathophysiology of epilepsy. Our previous research revealed that truncated TrkB receptors are upregulated in a rat hippocampal neuronal model of SREDs. In contrast, full-length TrkB receptors are downregulated. Furthermore, the activation of full-length TrkB signaling is suppressed by the overexpression of truncated TrkB. In this study, to regulate the expression of truncated TrkB receptor and full-length TrkB signaling, rno-miR-185-3p was transduced into the SREDs model. Then, the changes in the activity of L-type voltage-gated calcium channels (VGCCs) and in epileptiform discharges were investigated. Transduction of rno-miR-185-3p downregulated the expression of truncated TrkB and dramatically activated full-length TrkB signaling in the model. Next, we found that the activation of full-length TrkB signaling decreased the maximal Ca²⁺ current density in the model, delayed the steady-state activation and accelerated the inactivation of L-type VGCCs. Finally, the epileptiform discharges in the model could be impaired. Based on the above results, we suggest that the activation of full-length TrkB signaling may suppress the properties of L-type VGCCs, and thus ameliorate the epileptiform discharges in the model. The activation of full-length TrkB signaling may affect the inhibition of epilepsy. This provides a rationale for the activation of full-length TrkB signaling in preventive therapies.

The Suppression of Epileptiform Discharges in Cultured Hippocampal Neurons Is Regulated via Alterations in TrkB.FL Signaling Activity

Hongjun Huo

Baotou Medical College, Baotou, 104060, Inner Mongolia Autonomous Region, China.

Abstract:

Epilepsy is a common neurological disease. Understanding the mechanisms of epileptogenesis at the cellular and molecular levels may provide novel targets for preventing this disorder. Brain-derived neurotrophic factor (BDNF) and its receptor tropomyosin-related kinase type B (TrkB) are believed to be critical for epileptogenesis. Previous studies have revealed possible changes in the expression of full-length TrkB receptors (TrkB.FL) and truncated TrkB receptors (TrkB.T) in neurodegenerative disorders. In this study, we investigated alterations in TrkB receptor expression and TrkB signaling activity in a rat hippocampal neuronal model of spontaneous recurrent epileptiform discharges (SREDs) and the effects on the epileptiform discharges. To induce epileptiform discharges, we established a model with Mg²⁺-free treatment. We found a dramatic upregulation of TrkB.T and a decrease in TrkB.FL in the SREDs model. Calpain contributed to the downregulation of TrkB.FL. The upregulation of TrkB.T required transcription and translation activity. Furthermore, BDNF induced the activation of TrkB.FL signaling. However, TrkB.FL signaling was inhibited in the SREDs model. Although calpain inhibitors prevented a decrease in TrkB.FL, they did not restrain the downregulation of TrkB.FL signaling activity in the model. However, a SREDs model with a translation inhibitor prevented the increase in TrkB.T and re-activated TrkB.FL signaling activity. Finally, we used electrophysiology to observe that a downregulation of TrkB.T could relieve the representative epileptiform discharges in the model. These results, taken together, demonstrate that alterations in TrkB.FL signalling may be regulated via TrkB.T receptors. Upregulation of TrkB.FL signalling suppresses epileptiform discharges in the SREDs model.

Auditory guidance of imagined movements: Effects of real-time auditory feedback (sonification) guided mental imagery on knee proprioception

Shashank Ghai*, 1Gerd Schmitz, 1Tong-Hun Hwang, 1Alfred O. Effenberg

Public advocacy as a crucial part of longevity R&D and therapy

Shashank Ghai*, ¹Gerd Schmitz, ¹Tong-Hun Hwang, ¹Alfred O. Effenberg