scholarly journals Stoichiometric optimization of Gata4, Hand2, Mef2c, and Tbx5 expression for contractile cardiomyocyte reprogramming

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Zhentao Zhang ◽  
Wenhui Zhang ◽  
Young-Jae Nam
Keyword(s):  
Basic Tool ◽  
Future Studies ◽  
Sarcomeric Protein ◽  
Protein Levels ◽  
Tbx5 Expression ◽  
Low Efficiency ◽  
Tool Set ◽  
Vector Transduction

Abstract Reprogramming of fibroblasts to induced cardiomyocyte-like cells (iCMs) offers potential strategies for new cardiomyocyte generation. However, a major challenge of this approach remains its low efficiency for contractile iCMs. Here, we showed that controlled stoichiometric expression of Gata4 (G), Hand2 (H), Mef2c (M), and Tbx5 (T) significantly enhanced contractile cardiomyocyte reprogramming over previously defined stoichiometric expression of GMT or uncontrolled expression of GHMT. We generated quad-cistronic vectors expressing distinct relative protein levels of GHMT within the context of a previously defined splicing order of M-G-T with high Mef2c level. Transduction of the quad-cistronic vector with a splicing order of M-G-T-H (referred to as M-G-T-H) inducing relatively low Hand2 and high Mef2c protein levels not only increased sarcomeric protein induction, but also markedly promoted the development of contractile structures and functions in fibroblasts. The expressed Gata4 and Tbx5 protein levels by M-G-T-H transduction were relatively higher than those by transductions of other quad-cistronic vectors, but lower than those by previously defined M-G-T tri-cistronic vector transduction. Taken together, our results demonstrate the stoichiometric requirement of GHMT expression for structural and functional progresses of cardiomyocyte reprogramming and provide a new basic tool-set for future studies.

The orexin system: a potential player in the pathophysiology of absence epilepsy?

2021 ◽  
Vol 19 ◽  
Author(s):  
Roberta Celli ◽  
Gilles Van Luijtelaar
Keyword(s):  
Somatosensory Cortex ◽  
Molecular Mechanisms ◽  
Recent Literature ◽  
Absence Epilepsy ◽  
Experimental Animals ◽  
Future Studies ◽  
Protein Levels ◽  
System A ◽  
Cortical Circuit

Background : Absence epilepsy is characterized by the presence of spike-and-wave discharges (SWDs) at the EEG generated within the cortico-thalamo-cortical circuit. The molecular mechanisms involved in the pathophysiology of absence epilepsy are only partially known. WAG/Rij rats older than 2-3 months develop spontaneous SWDs, and they are sensitive to anti-absence medications. Hence, WAG/Rij rats are extensively used as a model for absence epilepsy with predictive validity. Objective : To examine the possibility that the orexin system, which supports the wake status in experimental animals and humans, plays a role in the pathophysiology of absence seizures. Methods : The perspective grounds its method on recent literature along with measurements of orexin receptor type-1 (OX1) protein levels in the thalamus and somatosensory cortex of WAG/Rij rats and non-epileptic Wistar control rats at two ages (25 days and 6-7 months). OX1 protein levels were measured by immunoblotting. Results : The analysis of the current literature suggests that the orexin system might be involved in the pathophysiology of absence epilepsy and might be targeted by therapeutic intervention. Experimental data are in line with this hypothesis showing that OX1 protein levels were reduced in the thalamus and somatosensory cortex of symptomatic WAG/Rij rats (6-7 months of age) with respect to non-epileptic controls, whereas these differences were not seen in pre-symptomatic, 25 days-old WAG/Rij rats. Conclusions : This might pave the way to future studies on the involvement of the orexinergic system in the pathophysiology of SWDs associated with absence epilepsy and its comorbidities.

scholarly journals Limitations to intergenerational inheritance: subchronic paternal stress preconception does not influence offspring anxiety

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
K. A. Fennell ◽  
R. G. G. Busby ◽  
S. Li ◽  
C. Bodden ◽  
S. J. Stanger ◽  
...  
Keyword(s):  
Low Dose ◽  
Sperm Count ◽  
Body Weight Gain ◽  
Epigenetic Inheritance ◽  
Receptor Protein ◽  
Future Studies ◽  
Protein Levels ◽  
Plus Maze ◽  
History Of ◽  
Paternal Stress

Abstract Independent studies have observed that a paternal history of stress or trauma is associated with his children having a greater likelihood of developing psychopathologies such as anxiety disorders. This father-to-child effect is reproduced in several mouse models of stress, which have been crucial in developing a greater understanding of intergenerational epigenetic inheritance. We previously reported that treatment of C57Bl/6J male breeders with low-dose corticosterone (CORT) for 28 days prior to mating yielded increased anxiety-related behaviours in their male F1 offspring. The present study aimed to determine whether subchronic 7-day CORT treatment of male mice just prior to mating would be sufficient to induce intergenerational modifications of anxiety-related behaviours in offspring. We report that subchronic CORT treatment of male breeders reduced their week-on-week body weight gain and altered NR3C1 and CRH gene expression in the hypothalamus. There were no effects on sperm count and glucocorticoid receptor protein levels within the epididymal tissue of male breeders. Regarding the F1 offspring, screening for anxiety-related behaviours using the elevated-plus maze, light–dark box, and novelty-suppressed feeding test revealed no differences between the offspring of CORT-treated breeders compared to controls. Thus, it is crucial that future studies take into consideration the duration of exposure when assessing the intergenerational impacts of paternal health.

A basic tool set for a generalized directional model

Public Choice ◽  
2009 ◽  
Vol 140 (1-2) ◽  
pp. 85-104 ◽  
Author(s):  
Eric Linhart ◽  
Susumu Shikano
Keyword(s):  
Basic Tool ◽  
Directional Model ◽  
Tool Set

scholarly journals Spectra: a specification language for reactive systems

2021 ◽  
Author(s):  
Shahar Maoz ◽  
Jan Oliver Ringert
Keyword(s):  
Reactive Systems ◽  
Specification Language ◽  
Lessons Learned ◽  
Science Students ◽  
Future Studies ◽  
Reactive Synthesis ◽  
Quality Specifications ◽  
Software Engineers ◽  
Tool Set ◽  
Potential Use

AbstractWe introduce Spectra, a new specification language for reactive systems, specifically tailored for the context of reactive synthesis. The meaning of Spectra is defined by a translation to a kernel language. Spectra comes with the Spectra Tools, a set of analyses, including a synthesizer to obtain a correct-by-construction implementation, several means for executing the resulting controller, and additional analyses aimed at helping engineers write higher-quality specifications. We present the language in detail and give an overview of its tool set. Together with the language and its tool set, we present four collections of many, non-trivial, large specifications, written by undergraduate computer science students for the development of autonomous Lego robots and additional example reactive systems. The collected specifications can serve as benchmarks for future studies on reactive synthesis. We present the specifications, with observations and lessons learned about the potential use of reactive synthesis by software engineers.

scholarly journals APOE4 Copy Number-Dependent Proteomic changes in the Cerebrospinal Fluid

2020 ◽  
Author(s):  
Miles Berger ◽  
Mary Cooter ◽  
Alexander S. Roesler ◽  
Stacey Chung ◽  
John Park ◽  
...  
Keyword(s):  
Copy Number ◽  
Disease Risk ◽  
Clinical Status ◽  
Future Studies ◽  
Complement Proteins ◽  
Protein Levels ◽  
Proteomic Data ◽  
False Discovery ◽  
Significant Expression ◽  
Lower Expression

AbstractBackgroundAPOE4 has been hypothesized to increase Alzheimer’s disease risk by increasing neuroinflammation, though the specific neuroinflammatory pathways involved are unclear.ObjectivesTo characterize CSF proteomic changes as a function of APOE4 copy number.MethodsWe analyzed targeted proteomic data obtained on ADNI CSF samples using a linear regression model adjusting for age, sex, and APOE4 copy number, and a second linear model also adjusting for AD clinical status. False Discovery Rate (FDR) was used to correct for multiple comparisons.ResultsIn the first model, increasing APOE4 copy number was associated with significant expression decreases in a CRP peptide (q=0.006), and significant expression increases in peptides from ALDOA, CH3L1 (YKL-40), and FABPH (q<0.05 for each). In the second model (controlling for age, sex, and AD clinical status), increasing APOE4 copy number was associated with significant expression decreases in a CRP peptide (q=0.009). In both models, increased APOE4 copy number was associated with trends towards lower expression of all 24 peptides from all 8 different complement proteins measured here, although none of these differences were statistically significant. The odds of this happening by chance for 24 unrelated peptides would be less than 1 in 16 million.ConclusionsIncreasing APOE4 copy number was associated with decreased CSF CRP levels and increased CSF ALDOA, CH3L1 and FABH levels; the CRP decrease remained significant after controlling for AD clinical status. Increased APOE4 copy number may also be associated with decreased CSF complement pathway protein levels, a hypothesis for investigation in future studies.

scholarly journals TRPV1 Responses in the Cerebellum Lobules VI, VII, VIII Using Electroacupuncture Treatment for Chronic Pain and Depression Comorbidity in a Murine Model

2021 ◽  
Vol 22 (9) ◽  
pp. 5028
Author(s):  
Bernice Lottering ◽  
Yi-Wen Lin
Keyword(s):  
Chronic Pain ◽  
Patient Outcomes ◽  
Sham Group ◽  
Future Studies ◽  
Protein Levels ◽  
Voltage Gated Sodium Channels ◽  
Electroacupuncture Treatment ◽  
Subtype 3

Depression is a prominent complex psychiatric disorder, usually complicated through expression of comorbid conditions, with chronic pain being among the most prevalent. This comorbidity is consistently associated with a poor prognosis and has been shown to negatively impact patient outcomes. With a global rise in this condition presenting itself, the importance of discovering long-term, effective, and affordable treatments is crucial. Electroacupuncture has demonstrated renowned success in its use for the treatment of pain and is a widely recognized therapy in clinical practice for the treatment of various psychosomatic disorders, most notably depression. Our study aimed to investigate the effects and mechanisms of Acid-Saline (AS) inducing states of chronic pain and depression comorbidity in the cerebellum, using the ST36 acupoint as the therapeutic intervention. Furthermore, the role of TRPV1 was relatedly explored through the use of TRPV1−/− mice (KO). The results indicated significant differences in the four behavioral tests used to characterize pain and depression states in mice. The AS and AS + SHAM group showed significant differences when compared to the Control and AS + EA groups in the von Frey and Hargreaves’s tests, as well as the Open-Field and Forced Swimming tests. This evidence was further substantiated in the protein levels observed in immunoblotting, with significant differences between the AS and AS + SHAM groups when compared to the AS + EA and AS + KO groups being identified. In addition, immunofluorescence visibly served to corroborate the quantitative outcomes. Conclusively these findings suggest that AS-induced chronic pain and depression comorbidity elicits changes in the cerebellum lobules VI, VII, VIII, which are ameliorated through the use of EA at ST36 via its action on TRPV1 and related molecular pathways. The action of TRPV1 is not singular in CPDC, which would suggest other potential targets such as acid-sensing ion channel subtype 3 (ASIC3) or voltage-gated sodium channels (Navs) that could be explored in future studies.

scholarly journals APOE4 Copy Number-Dependent Proteomic Changes in the Cerebrospinal Fluid

2020 ◽  
pp. 1-20
Author(s):  
Miles Berger ◽  
Mary Cooter ◽  
Alexander S. Roesler ◽  
Stacey Chung ◽  
John Park ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Copy Number ◽  
Linear Models ◽  
Disease Risk ◽  
Clinical Status ◽  
Future Studies ◽  
Protein Levels ◽  
Proteomic Data ◽  
False Discovery ◽  
The One

Background: APOE4 has been hypothesized to increase Alzheimer’s disease risk by increasing neuroinflammation, though the specific neuroinflammatory pathways involved are unclear. Objective: Characterize cerebrospinal fluid (CSF) proteomic changes related to APOE4 copy number. Methods: We analyzed targeted proteomic data from ADNI CSF samples using a linear regression model adjusting for age, sex, and APOE4 copy number, and additional linear models also adjusting for AD clinical status or for CSF Aβ, tau, or p-tau levels. False discovery rate was used to correct for multiple comparisons correction. Results: Increasing APOE4 copy number was associated with a significant decrease in a CRP peptide level across all five models (q <  0.05 for each), and with significant increases in ALDOA, CH3L1 (YKL-40), and FABPH peptide levels (q <  0.05 for each) except when controlling for AD clinical status or neurodegeneration biomarkers (i.e., CSF tau or p-tau). In all models except the one controlling for CSF Aβ levels, though not statistically significant, there was a consistent inverse direction of association between APOE4 copy number and the levels of all 24 peptides from all 8 different complement proteins measured. The odds of this happening by chance for 24 unrelated peptides would be less than 1 in 16 million. Conclusion: Increasing APOE4 copy number was associated with decreased CSF CRP levels across all models, and increased CSF ALDOA, CH3L1, and FABH levels when controlling for CSF Aβ levels. Increased APOE4 copy number may also be associated with decreased CSF complement pathway protein levels, a hypothesis for investigation in future studies.

Educating the educators to innovate: the need to reinvent academia's mission and to reengineer their basic tool: the Educator

2013 ◽  
Vol 1 (1) ◽  
pp. 7-10
Author(s):  
José Tribolet
Keyword(s):  
Learning Environments ◽  
Social Value ◽  
Human Being ◽  
Academic Institutions ◽  
Basic Tool ◽  
Academic Activities ◽  
The World ◽  
Tool Set

Innovating is an attitude towards the world.For humanity to improve its capabilities to innovate, then innovation, as an individual capability with social value, must be nurtured since birth, in every human being, along with the full toolset of values, such as liberty, responsibility, solidarity, compassion, honesty, among others. A foundation for the acquisition and maturation of the innovation attitudes in an individual lay on its basic drive to question the world, to understand, i.e. model it, and in such process, to identify perplexities that confront him as problems to be solved or opportunities to be addressed. I consider the development and maturation of these capabilities in each individual an essential responsibility of academic institutions. To adopt encompassing processes for providing learning environments, for the students to acquire and develop innovation capabilities, Academic Institutions need to refurbish their production tool set, so that it acquires inherent innovation capabilities in all the dimensions of the academic activities. This is perhaps the biggest challenge today for these Institutions, because it requires massive reprogramming of the Professors mindsets and practices.

scholarly journals Increased Systemic HSP70B Levels in Spinal Muscular Atrophy Infants

2020 ◽  
Author(s):  
Eric J. Eichelberger ◽  
Christiano R. R. Alves ◽  
Ren Zhang ◽  
Marco Petrillo ◽  
Patrick Cullen ◽  
...  
Keyword(s):  
Spinal Muscular Atrophy ◽  
Therapeutic Response ◽  
Muscular Atrophy ◽  
Sequencing Analysis ◽  
Life Changes ◽  
Circulating Biomarkers ◽  
Future Studies ◽  
Protein Levels ◽  
Molecular Therapies

AbstractDespite newly available treatments for spinal muscular atrophy (SMA), novel circulating biomarkers are still critically necessary to track SMA progression and therapeutic response. To identify potential biomarkers, we performed whole-blood RNA sequencing analysis in SMA type 1 subjects under 1 year old and age-matched healthy controls. Our analysis revealed the Heat Shock Protein Family A Member 7 (HSPA7) as a novel candidate biomarker to track SMA progression early in life. Changes in circulating HSPA7 protein levels were associated with changes in circulating neurofilaments levels in SMA newborns and infants. Future studies will determine whether HSPA7 levels respond to molecular therapies.

Abstract 103: TBX20 Activates Cardiac Maturation Gene Programs Promoting Direct Human Cardiac Reprogramming

2020 ◽  
Vol 127 (Suppl_1) ◽  
Author(s):  
Yang Zhou ◽  
Yawen Tang ◽  
Vladimir G Fast ◽  
Lianzhong Zhao ◽  
Rui Lu ◽  
...  
Keyword(s):  
Stem Cell Differentiation ◽  
Developed Countries ◽  
Heart Tissue ◽  
Poor Quality ◽  
Transcriptomic Analysis ◽  
Mortality And Morbidity ◽  
T Box ◽  
Sarcomeric Protein ◽  
Low Efficiency ◽  
Computational Analyses

Direct cardiac reprogramming to generate induced cardiomyocyte like cells (iCMs) from fibroblasts has emerged as a promising therapeutic strategy for the treatment of heart failure, which is still the leading cause of mortality and morbidity in developed countries. While much is known regarding iCMs generated from mouse cells, the adaptation of direct cardiac reprogramming to human cells is hurdled with low efficiency and poor quality because of intrinsic differences between species. Recently, our single cell transcriptomic analysis during human direct cardiac reprogramming demonstrated the immature features of human iCMs (hiCMs) compared to endogenous cardiomyocytes (CMs) at the transcriptome level, suggesting that key factors required for CM maturation remain unidentified. We then utilized computational analyses of genomic and epigenomic data to identify the regulators underexpressed in hiCMs comparing to CMs derived from human heart tissue or stem cell differentiation. Among them, the top hit is T-box family transcription factor TBX20, whose role in direct cardiac reprogramming remains to be explored. Supplementing TBX20 in reprogramming cocktails, we found significantly increased sarcomeric protein expression, like αMHC and αActinin in hiCMs and the formation of well-organized myofibrils. TBX20’s effect has also been supported by transcriptomic analysis, where we found TBX20 activates a large number of underexpressed genes associated with sarcomere structure and ion channels, such as MYH7 , MYL2 , MYBPC3, SCN5A and KCNQ1 , suggesting that TBX20 likely enhances maturation and contractility of hiCMs. We also found that TBX20 cannot replace any of the reprogramming factors, suggesting that its potential role is complimentary to and interacting with the current cocktail. Moreover, known TBX20 targets showed inconsistent alteration in TBX20-transduced hiCMs, suggesting its novel regulation in a direct reprogramming setting. These findings assign a novel reprogramming factor TBX20 playing an essential role in generating mature hiCMs by establishing gene programs associated with cardiac muscle contractility.

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