bluuebunny/crossref_metadata_2025_split huggingface dataset:
Data from single nuclei RNA‐sequencing reveals a prodromal gene network response in excitatory neurons of a humanized rat Alzheimer’s disease model
Authors: Tipton Allison E , George Jonique , Ratner Marcia , Farb David , Russek Shelley 
Published: December 2021

AbstractBackgroundDespite significant investment into treatments for AD, little progress has been made in bringing new therapeutics to market. Recent failure of monoclonal antibodies targeting Ab plaques to slow disease progression in phase 3 clinical trials, suggests now more than ever it is imperative to identify early disease signatures before symptom onset whose modification may alter disease trajectory. Our collaborating lab recently identified early dysfunction in the transgenic tri‐synaptic hippocampal circuit using in vivo recordings during awake immobility in the TgF344‐AD rat model (doi: 10.1101/2021.05.10.443512). This dysfunction emerges between 6‐9 months, prior to when TgF344‐AD rat shows overt pathology (including Ab plaques, frank neuronal loss, and endogenous tau pathology) at 16 and 26 mo, but not at 6 mo (Towne, 2013).MethodsThe molecular basis of this trisynpatic circuit dysfunction was explored using 10x Genomics’ v3 gene expression assay to perform snRNA‐seq on freshly dissected hippocampi from 6 mo male and female littermate pairs of Tg and Wt rats (n=16). ∼2000 cells/subject were collected, and cDNA libraries were sequenced (NovaSeq 600) to a depth of ∼120k reads/nuclei. Cell Ranger Pipeline demultiplexed and aligned reads to rat pre‐mRNA reference and generated gene count matrices. A final dataset of 33,079 high quality nuclei were clustered (Louvain graph‐based), and cell‐type specific marke used to determine identity. Identity was collapsed into broad neuronal and glial cell types for differential gene expression (DEG) across genotype (Hurdle Model).ResultsDEGs revealed profound alterations between genotype in cell types important in the trisynaptic circuit, including dentate granule cells (DGCs), and excitatory neurons (ExcNs). (FDR cutoff=0.05, 1111 (DGCs) and 1070 (ExcN’s) DEGs between genotypes). Pathway enrichment revealed dysregulation relevant to memory function: including decreased synaptogenesis, neurite branching, proliferation, and microtubule and cytoskeleton dynamics. DEG’s were enriched for neurological signs including epilepsy, seizures, hyperactivity, and dementia.ConclusionPrior to development of overt pathology or memory deficits, TgF344‐AD rats display transcriptomic alterations in their excitatory neurons, suggestive of impaired ability to form new connections and susceptibility to aberrant, hyperactivity. Electrophysiological detection, followed by intervention targeting these early alterations in gene expression may prevent progression of disease to a symptomatic stage.
The short term continuous darkness affects brain and behavior in a sex‐dependent manner: A comparative study in C57BL/6 male and female mice
Authors: Singh Dhyanendra 
Published: December 2021

AbstractBackgroundStandard light‐dark cycle (12h light and 12h dark) has an imperative role in keeping experimental animals healthy and adaptive. If it gets perturbed, it can negatively impact the brain, behavior, and Physiology. The circadian clock (central clock) of hypothalamic suprachiasmatic nuclei received light signals from the environment. Subsequently, in collaboration with the peripheral clocks of the tissues, the central clock transformed these light signals into oscillations in behavior and physiology to maintain homeostasis in coping with environmental fluctuations or stressors. To understand how light‐dark cycle disruption affects the brain and, as a result, behavior, it's critical to simultaneously look at the molecular and behavioral changes. Further, sex is one of the inevitable variables neglected in neurobiology and behavioral study even after the advancement of science in the 20th centuryMethodIn the current study, we evaluated the influence of the altered light‐dark cycle on the brain (prefrontal cortex), stress response, and behavior of C57BL/6 male and female mice by exposing them to the continuous darkness (0h light and 24h dark) for three weeksResultOur results show that constant darkness leads to the change in behavior (in the open field test), the prefrontal cortex (pro‐ and anti‐inflammatory genes and neurotrophic factors), and stress response (stress hormones) in a sex‐dependent mannerConclusionThe present study laid the foundation for understanding how the altered light‐dark cycle perturbed behavior, brain, and stress response in a sex‐dependent way.
Intra‐ and inter‐rater reliability of brain's activation area measurement on imagej: An easy and low‐cost method for cognitive and neurophysiological studies
Authors: Monteiro‐Junior Renato Sobral , Santos Iaggo Raphael David , Machado Frederico Sander ,   Research and Study Group on Neuroscience, Exercise, Health and Sport ‐ GENESEs
Published: December 2021

AbstractBackgroundNeuroimage is an important resource in cognitive neuroscience1 but its analysis is complex and requires reliable and affordable procedures. The present study sought to determine the intra‐ and inter‐rater reliability of brain´s activation area of neuroimages obtained derived from an integrated EEG system equipment.MethodThe study follows the Declaration of Helsinki. Fifteen cerebral cortex images were obtained through Neurometry v6 (Bioevolution®, Brazilian National Health Surveillance Agency registration 81403519002)2,3 from 15 individuals above 60 years, who consented to participate of the study. This device consists of a 19‐channel EEG (10‐20 system). Amplitude of brain waves (delta, theta, alpha, beta, and gamma) are automatically processed, filtered and analyzed with Math Library MtxVec, LAPACK and Math Kernel BLAS with codes Streaming SIMD (SSE2, SSE3 and SSE4)2. The neuroimages display different colors according to the amplitude of brain waves: brown/blue (low activation), green/yellow (moderate activation), and orange/red (high activation)2. Images retrieved were exported and analyzed with the ImageJ software (National Institutes of Health, Bethesda, Maryland, USA) in 3 phases: 1st An experienced researcher (RSMJ) standardized Broadmann´s area as the region of interest (ROI); 2nd The researcher specified the brain´s activation area in accordance with colored regions (brown, blue, green, yellow, orange, and red) in the Broadmann´s area 10 (BA10) covering 1,391 pixels. Colors were covered using 2 ImageJ tools (polygon and freehand selections). In order to calculate the colored area of interest, pixels quantification was performed with a 400% magnification filter (Figure 1). The procedure was repeated 24h later for the intra‐rater reliability; 3rd An inexperienced assessor (IRDS) independently performed the same procedure of the 2nd phase. Intraclass Correlation Coefficients (ICC3,1 and ICC3,k)4 were calculated to determine the intra‐ and inter‐rater reproducibility. Typical Error of Measurement (TEM=SD∑difference between measures÷√2)5 was also calculated to quantify typical deviations from the right measure.ResultsIntra‐ and inter‐rater reliability were excellent (ICC=0.99, p<0.01) and good (ICC=0.82, p<0.01), respectively (Table 1). TEM varied from 4 to 17%.ConclusionQuantification of the brain´s activation area of neuroimages derived from Neurometry v6 using the ImageJ ROI is highly reliable, independently of the assessor´s experience.
Evaluation and optimization of different cf‐RNA extraction methods for Alzheimer‐related genes detection
Authors: Le Hoang Anh P , Tran Tai T , Cao Minh Thien H , Le Mai Thao T , Pham Bao H , Ha Huong T T 
Published: December 2021

AbstractBackgroundCell‐free RNA (cf‐RNA) in plasma has become a fascinating less‐invasive diagnostic approach for Alzheimer’s Disease (AD) since brain‐originated mRNA and microRNA can enter the bloodstream through the blood‐brain barrier. Studying cf‐RNA expression in blood samples collected from AD‐patients and at‐risks patients allows further understanding of disease pathogenesis, contributes to the development of early diagnosis and prognosis methods. However, contaminants risks, low yield, and integrity can alter final cf‐RNA quality and concentration, challenging their application. This study aims to evaluate and optimize different cf‐RNA extraction protocols, as well as to detect the AD‐related transcripts in the final sample.MethodWe assessed different cf‐RNA extraction methods and evaluated hemolysis ‐ the main contamination threat to the final product. Blood samples were collected from healthy volunteers into designated collection tubes to compare their effects on hemolysis and RNA products. Plasma cf‐RNA was extracted using the Trizol and Zymo cf‐RNA kits, with two modifications to the Trizol protocol: addition of glycogen and with the use of Rneasy mini kit to increase cf‐RNA yield. Among 30 blood samples were collected, 12 were treated with Zymo, six with Trizol, eight with Trizol w/glycogen and four with Rneasy. cf‐RNA was then converted to cDNA and examined with PCR and qPCR.ResultNorgen tubes showed a lower hemolysis level compared to Streck tubes. The yield of cf‐RNA from the Zymo kit was about 5‐18 µg/ml. The Trizol protocol resulted in almost no detectable cf‐RNA. The addition of glycogen significantly increased the yield to a comparable level with the Zymo kit (4‐15 µg/ml). No genomic DNA contamination was detected. Despite high cf‐RNA yield from the modified Trizol protocol, the housekeeping gene Beta‐actin (ACTB) or other AD‐related transcripts (CNTF and MAG) could not be detected. Using cDNA obtained from the Zymo kit, ACTB was reliably detected, but AD‐related transcripts’ results varied. cDNA concentration was high‐correlated with the melt curve analysis of ACTB qPCR, suggesting Zymo kit obtained cDNA had high integrity.ConclusionOverall, the Zymo kit is superior to the Trizol‐based protocols in both cf‐RNA yield and integrity. The final sample showed detectable levels of ACTB and AD‐related transcripts.
Dissecting a neuron‐to‐liver crosstalk to modulate lipid metabolism in lisosomal storage diseases
Authors: Garcia‐Macia Marina , Bolaños Juan Pedro 
Published: December 2021

AbstractBackgroundNeuronal ceroid lipofuscinoses (NCL), known as Batten disease, are the most common of the rare neurodegenerative disorders in children. To date, defects in thirteen different genes have been identified in NCL patients. Despite the genetic heterogeneity, Batten diseases are grouped together based on clinical similarities and broadly uniform neuropathological features, including accumulation of lipofuscin in lysosomes, as well as profound neurodegeneration and widespread gliosis. Amongst these, the incidence of Cln7 disease, caused by mutation in MFSD8 gene, is the highest in southern and Mediterranean Europe. CLN7/MFSD8 encodes a lysosomal membrane glycoprotein with unknown function. Lysosomes are the only organelles able to hydrolyse triacylglycerols, which fuels the mitochondria for energy generation. The autophagic machinery provides TGs to the lysosomes through a process known as lipophagy. Although defective autophagy has been related with Cln7 disease, Cln7 role in lipid metabolism is unknown, particularly in lipophagy. Here, we hypothesized that disruption of lipophagy links neuronal death and Cln7‐mediated Batten disease.MethodTo address this, we have studied lipophagy in Cln7 knockout (Cln7‐KO) mice and investigated whether Cln7 loss in the hypothalamus disrupts liver lipophagy. We have obtained experimental data from different techniques in Cln7‐KO mice fibroblasts, liver, brown fat and brain, to stablish a connection between the brain and the metabolism of the peripheral tissues.ResultThus, our data show that Cln7 deficiency in the hypothalamus damages liver lipophagy resulting in fat accumulation. Ongoing work is being developed to validate these observations using robust metabolic and in vivo uncoupling approaches.ConclusionCln7 deficiency causes neuronal damages which seem to produce lipophagy impairment in the periphery tissues.
In Vitro and In Vivo Pharmacological Comparison of Mu‐Opioid Receptor Activity of the Kratom (Mitragyna speciosa) Alkaloid Mitragynine and its Metabolite 7‐Hydroxymitragynine
Authors: Calvache Maria P Guerrero , Obeng Samuel , Leon Francisco , Gamez‐Jimenez Lea R , Patel Avi , Ho Nicholas P , Crowley Morgan L , Pallares Victoria , Mottinelli Marco , McCurdy Christopher R , McMahon Lance R , Hiranita Takato 
Published: December 2021

AbstractBackgroundThe µ‐opioid receptor (MOR) pharmacology of mitragynine and its 7‐hydroxy metabolite have not been fully established.MethodFor both we assessed binding affinity and efficacy in vitro, as well as discriminative stimulus and antinociceptive effects in rats.ResultBoth mitragynine (MG) and 7‐hydroxymitragynine (7‐MG) had higher binding affinities at human MOR (Ki values 77.9 and 709 nM, respectively) than at the human k‐ (KOR) or d‐opioid receptors (DOR). MG was identified as a MOR antagonist while 7‐MG was identified as a MOR partial agonist (Emax=41%) using the [35S]GTPγS functional assay. In Sprague Dawley rats trained to discriminate 3.2 mg/kg morphine from vehicle (i.p.), MG produced a maximum of 72% morphine‐lever responding. Whereas morphine produced a maximum of 65% MG‐lever responding in a separate group of rats trained to discriminate 32 mg/kg MG from vehicle (i.p.), other MOR agonists (i.p.) produced high percentages of morphine‐lever and MG‐lever responding (respectively): 7‐MG (100% and 98%), fentanyl (100% and 80%), buprenorphine (100% and 79%), and nalbuphine (99% and 98%). MG (up to 56 mg/kg, i.p.) did not produce significant antinociception, whereas 7‐MG (17.8 mg/kg, i.p.) produced robust antinociception in the hotplate assay with the temperature set at 52 °C. Naltrexone (0.032 mg/kg, i.p.) antagonized the discriminative stimulus, rate‐decreasing, and antinociceptive effects of morphine and 7‐MG. Conversly, naltrexone (0.032 mg/kg, i.p.) antagonized the discriminative stimulus effects of MG, but not its rate‐decreasing effects. Pretreatment with MG (5.6 mg/kg, i.p.) potentiated the discriminative‐stimulus effects of morphine but antagonized the antinociceptive effects of morphine.ConclusionStriking differences in MOR efficacy in vitro are consistent with 7‐MG having greater MOR efficacy than MG in vivo. MG exerts both MOR antagonist and agonist activity depending on the MOR efficacy required.
Exercise‐induced metabolic changes in astrocytes in the 5xFAD mouse model of Alzheimer’s Disease
Authors: Rosolen Sarah Alyssa 
Published: December 2021

AbstractBackgroundAlzheimer’s Disease (AD), a progressive neurodegenerative disease and the second most common cause of death in Australia (ABS 2019), is an increasingly prominent societal issue, exacerbated by an ageing population and the absence of effective disease modifying treatments. Focus has shifted to targeting modifiable risk factors to slow or prevent AD onset and progression, such as exercise, diet and other lifestyle factors. Adiponectin, an anti‐inflammatory adipokine, regulates energy metabolism and is associated with metabolic pathways potentiated following exercise. Animal and human studies involving both healthy individuals and those with metabolic dysfunction, have consistently demonstrated increased serum adiponectin levels following various exercise regimes. The ability of adiponectin to cross the blood brain barrier and thus mediate neuronal metabolism is contentious. However, the expression of adiponectin receptors within neuronal cells implies adiponectin holds an important function within the brain.MethodsWe investigated the cortical expression of adiponectin receptors, AdipoR1 and AdipoR2, in the aged 5XFAD mouse model of AD following an exercise intervention. Immunohistochemical techniques were applied to double stain brain tissue for adiponectin receptor and astrocyte expression.ResultsWe observed neuronal AdipoR1 and AdipoR2 expression throughout the cortex of both exercised and sedentary control mice, as well as extensive expression of activated astrocytes.ConclusionsColocalisation analysis suggests astrocytes may utilise adiponectin receptors to fuel their metabolic activity in degrading toxic amyloid plaques within the AD brain.
Increased Plasma brain‐derived neurotrophic factor (BDNF) as a Biomarker for Differentiating Mild Cognitive Impairment from Cognitive Healthy: A case‐control study
Authors: Ng Ted Kheng Siang , Coughlan Christina M , Heyn Patricia C , Tagawa Alex , Carollo James , Kua Ee‐Heok , Mahendran Rathi 
Published: December 2021

AbstractBackgroundAcross the spectrum of Alzheimer’s disease (AD), we previously conducted a meta‐analysis and concluded significantly decreased brain‐derived neurotrophic factor (BDNF) in AD. However, there are inconsistent findings on peripheral BDNF levels in mild cognitive impairment (MCI), even across different meta‐analyses, which have been attributed to the high heterogeneity caused by multiple clinical and laboratory factors. Thus, BDNF’s discriminative accuracy for identifying all‐cause MCI and its subtypes remains unknown.MethodsThis study, while accounting for heterogeneity, compared a healthy control cohort (n=56, 45 female) and an MCI cohort (n=40, 28 female) to determine whether plasma BDNF (pBDNF), hs‐CRP, and DHEA‐S can differentiate between healthy and MCI individuals, including the two MCI subtypes (amnestic [aMCI] and non‐amnestic [non‐aMCI]). Furthermore, the associations between the biomarkers and detailed neurocognitive tests were examined. Adults with cerebral palsy were included as sensitivity analyses.ResultCompared to healthy controls, only pBDNF was significantly increased in all‐cause MCI, aMCI, and non‐aMCI. Furthermore, pBDNF had good (AUC=0.84, 95% CI=0.74 to 0.95, p<0.001) and excellent discriminative accuracy (AUC=0.92, 95% CI=0.84 to 1.00, p<0.001) for all‐cause MCI and non‐amnestic MCI, respectively. pBDNF was also significantly and positively associated with plasma hs‐CRP (β=0.26, 95% CI=0.02 to 0.50, p=0.038), despite attenuated association upon controlling for BMI (β=0.15, 95% CI=‐0.08 to 0.38, p=0.186). Multiple associations with detailed neurocognitive tests were observed.ConclusionThese findings support increased pBDNF as a compensatory mechanism in preclinical dementia, and the neurotrophic and inflammatory hypotheses of cognitive impairment.
Astrocytic GLUT1 ablation improves systemic glucose metabolism and promotes cognition
Authors: Ardanaz Carlos G , Gil María Javier Ramírez , Smerdou Cristian , Solas Maite 
Published: December 2021

AbstractBackgroundGlucose supply from the blood to the brain is controlled by the glucose transporter GLUT1, highly expressed in astrocytes, which coordinate brain glucose supply, metabolization and storage. Ablating GLUT1 at the blood‐brain barrier (BBB) endothelial cells leads to BBB breakdown, brain glucose hypometabolism and impaired cognition, but this approach cannot discriminate between insufficient glucose supply and BBB breakdown‐derived effects. Such question is the focus of the present work, which aims to elucidate the relevance of astrocytic GLUT1 to cellular, brain and systemic glucose metabolism, and to cognition.MethodsTo address these questions, GLUT1 was ablated from primary astrocytes. Cellular metabolism was examined using an extracellular flux analyzer (Seahorse). In vivo, astrocytic GLUT1 was ablated using a tamoxifen‐inducible Cre/LoxP approach (GLUT1ΔGFAP mice). 18F‐FDG PET, glucose and insulin tolerance and insulin secretion and fasting‐induced hyperphagia were characterized. BBB integrity was examined by vessel immunostaining and capillary‐depleted brain analysis. Recognition and spatial memory were assessed using Novel Object Recognition and Morris Water Maze tasks. To address the implication of purinergic signaling in those effects, a purinergic receptor antagonist (PPADS) was intracerebroventricularly administered before each behavioral test.ResultsGLUT1‐ablated astrocytes showed reduced glucose uptake and glycolysis, although preserving total ATP production. Unexpectedly, postnatal astrocytic GLUT1 deletion increased CNS glucose utilization. GLUT1ΔGFAP mice showed an improved metabolic status from which obese animals especially benefited. Specifically, GLUT1ΔGFAP mice were more efficient at suppressing hyperphagia and readjusting systemic glucose levels after hyperglycemia, exhibiting marked increase in insulin secretion. These effects were coupled with enhanced BAT activity, and reduced BAT adiposity. In parallel with this improved systemic homeostasis, GLUT1ΔGFAP mice performed both recognition and spatial memory tasks properly, even outperforming control mice. Noteworthy, those effects could be due to higher astrocytic ATP release. Indeed, central administration of PPADS could reverse improvements in metabolic and cognitive behaviors in mice with astrocyte GLUT1 knockout.ConclusionOverall, this study demonstrates that astrocytic GLUT1 ablation impairs astrocytic glucose availability but enhances brain glucose utilization, reprograms systemic glucose metabolism towards a more efficient glucose‐handling phenotype and promotes cognitive abilities, which could be a key factor in neurodegenerative diseases such as Alzheimer’s disease.
English Lessons with the Use of Songs for People with Mild Cognitive Impairment
Authors: Makri Marina , CHRISTAKIDOU ALEXANDRA , Tsolaki Magda 
Published: December 2021

AbstractBackgroundPeople suffering from Mild Cognitive Impairment (MCI) need to improve their cognitive abilities in order to avoid the deterioration of their condition. Learning a foreign language is a good way for cognitive enhancement. However, since these people have memory deficits and lack of attention abilities, conventional teaching methods cannot be used. The use of songs in foreign language lessons is proved to be a very good technique for more effective acquisition of the language while also improving well‐being and relaxation.MethodThe Erasmus+ funded “E.L.So.M.C.I” program is a 2 year pan‐European initiative that aims to develop an educational program on teaching English to people with MCI and using English songs as a main tool for the teaching process. The methodology of this educational program is based on innovative teaching approaches such as “Communicative Language Teaching” and “Natural Approach and the method of “Neuro‐linguistic Programming (NLP)”. These methods place great emphasis on verbal communication, creation of a positive environment in class, reduction of stress and encouragement of learners to learn step by step in a natural and pleasant way.ResultIn this moment, the project has entered its first operational phase, that has the objective of producing:‐ A Workshops’ Methodological Guide which is a document provides trainers with guidelines for teaching languages that focuses on communication in English (for the learners) and preparation of the lesson plan for each lesson. The Methodological Guide includes two kinds of materials, one for the participants‐students and one for the professionals (trainers).‐ Several Workshops for English learning, addressed to people with MCI (MMSE between 28 and 24).‐ Co‐Created Methodological Guide and Training for trainers via MOOC.ConclusionThis project and methodology can improve cognitive functions, prevent from Alzheimer’s disease, support the teaching process through the use of songs, reduce stress and increase positive emotions, improve participants’ socialization, as well as, develop the sense of belonging to a group.