bluuebunny/crossref_metadata_2025 huggingface dataset:
Neuroscience laboratory instruction using a computer‐based neuroanatomical learning tool (720.2)
Authors: Severson Arlen
Published: June 2021

An interactive computer‐based neuroanatomical learning tool is being used in an integrated neuroscience laboratory to facilitate learning neuroanatomical structures, relationships, significant morphological and functional features of the human central nervous system, and clinical relevance of the neuroanatomical structures. Gross images of intact and dissected brains, thick and stained thin brain sections, thin stained brainstem and spinal cord sections, and MR images are studied. Neural structures to be identified are listed on the right‐hand side of the computer screen. Clicking on the name of a specific structure produces a transparent overlay of that structure. Structures can be identified also by moving the pointer over the image and clicking on a specific structure. With either approach, the structure is identified by a colored overlay and a collapsible dialog box at the bottom on the screen that provides a brief description of the structure and its functional role in the nervous system. In addition, images from the program are incorporated into clinical vignettes to reinforce learning. Data from student surveys and comments indicate that this is an effective way to learn the anatomical and functional aspects of the nervous system that is available at the student’s convenience. The computer‐assisted learning tool accomplishes our laboratory goals and is enthusiastically accepted by the students.

Grant Funding Source: Supported by Minnesota Medical Foundation Herz Development Award
Incorporating bioinformatics into K‐12 Outreach activities (720.8)
Authors: Toby Inimary , Widmer Jonah
Published: June 2021

The use of biological databases has emerged as an exciting part of most research laboratories and programs. Until the turn of the decade, tasks such as searching genomes for differences between genes or transcription factors would have seemed daunting and perhaps laborious. However nowadays in the post‐genomic era, researchers all over the world can have easily access these various types of information. Many have suggested that we must find ways to continue inspiring our nation’s young people to develop a love or appreciation for science.

We wanted to exploit the availability of these databases to develop research projects aimed at students in grades 9th‐ 12th. This population of students can be challenging to teach because they are always looking for some practical applications to the subject learned and often have misapprehensions about how science is done. Our goal for the students research projects was to use Bioinformatics tools such as computer programming and database mining as a way to extract physiological information about microbial organisms. We utilized Pfam, signalp, CDART, and TmmHmm databases for analysis. Students were provided with protein sequences from various microorganisms for comparative analysis.

We found that these computer‐based projects provided a unique opportunity for the student to engage in research while testing their knowledge from basic biology such as examining differences amongst genes, proteins, and genomes. Students were also able to learn skills, including programming, statistical analysis, and MySQL database management. Upon completion of their projects, we reviewed the students’ responses from surveys and final reports. These enabled us to ascertain what the students gained from the internship, what they enjoyed and what if any, improvements were necessary as we offer this to more students. In the future, we would like to track the progress of these students as they return to high school and go on to pursue further studies in postsecondary institutions.

Grant Funding Source: NIH INBRE grant
Alzheimer’s is associated with neuropathological changes in olfactory bulb that are related to APOE genotype (728.1)
Authors: Parcon Paul , Aboud Orwa , Biedermann JoAnn , Mrak Robert , Griffin W. Sue
Published: June 2021

Alzheimer patients (AD) may experience changes in olfaction before clinical or neuropathological manifestations. Neuroinflammation, in particular interleukin‐1 (IL‐1), is known to induce each of the precursors of these manifestations.

We studied anatomical and pathological features in frozen and paraffin‐embedded tissue sections of olfactory bulbs resected from autopsy tissue of 15 patients (5 APOE ε3,3 neurologically normal, 5 APOE ε3,3 and 5 APOE ε4,4 AD patients) with an age range from 68 to 85 yrs, using fluorescent and western immunochemistry.

Compared to control, patients with AD had greater expression of hyper‐phosphorylated tau, particularly in those of APOE ε4,4 genotype compared to ε3,3. These findings were associated with increased expression of IL‐1, and the number of activated microglia was elevated. Furthermore, hyperphosphorylated tau expression was high in neuron cell bodies compared to that in axons in the olfactory tract. Although corpora amylacea were abundant, there was no disease‐related difference in distribution, number, or size.

Our findings are consistent with the idea that changes in expression of disease‐associated precursor proteins in olfactory structures and the cytokines that drive their expression are related to APOE genotype and are important elements in the neuroanatomical spread of the neuropathological changes in AD.

Grant Funding Source: Supported by NIH/NIA AG12411 and the Windgate Foundation
Fkbp5 and its interaction in fear extinction (728.15)
Authors: Criado‐Marrero Marangelie , Porter James
Published: June 2021

Introduction: The hypothalamic‐pituitary‐adrenal (HPA) axis and the limbic system are involved in the response to stressful situations. Disruption of this negative feedback loop has been related to depression and anxiety like behaviors such as Post‐traumatic Stress Disorders (PTSD). To prevent an excessive (pathological) stress response, the neuronal effects induced by the stress hormone corticosterone (or cortisol in humans) are modulated by negative feedback through the glucocorticoid receptors (GR) in neurons. Two key regulators of this negative intracellular feedback are the FK506‐binding protein 4 (Fkbp4) and the FK506‐binding protein 5 (Fkbp5). Deletion of the Fkbp5 gene is associated with less anxiety and variants of the Fkbp5 gene are associated with PTSD. The objective of this study is to examine the role of Fkbp5 during fear extinction learning. Hypothesis: Signaling via Fkpb5 and its interaction with glucocorticoid receptors in the prefrontal cortex plays an important role in fear extinction by modulating the neuronal effects of corticosterone. Methodology: Young and adult male rats were subjected to habituation, conditioning, and extinction learning phases. They were sacrificed and their prelimbic (PL) and infralimbic (IL) cortical tissue was extracted. Microarrays and RT‐ PCR were used to determine the expression of Fkbp4 and Fkbp5 mRNA during the different behavioral phases. Results: Fkbp5 mRNA expression, but not Fkbp4, in IL is reduced 5 hrs after cued fear extinction in rats. However, no changes were observed in PL. Conclusion: These results suggest that a reduction in Fkbp5 activity is necessary for consolidation of fear extinction and that fear extinction could be enhanced by reducing Fkbp5 expression in IL. Future Projections: By further examining the role of Fkbp5 in fear conditioning and extinction, we could identify novel targets for enhancing the treatment of PTSD patients.

Grant Funding Source: Supported by RISE grant R25GM082406, Molecular Biology Core Lab (Grant RR003050/MD007579),
Affordable assessment of newborn brain health following perinatal asphyxia in East Africa: a pilot study (728.6)
Authors: Clay David , Stojanoski Bobby , Linke Annika , Cameron Daniel , Wasunna Aggrey , Rulisa Stephen , Cusack Rhodri
Published: June 2021

Perinatal asphyxia is a major cause of infant mortality in the developing world with 280,000 deaths per year in sub‐Saharan Africa alone. Brain injuries resulting from asphyxia can have detrimental effects on brain function, which can negatively impact quality of life in those who survive. Since clinical manifestations are often subtle and the therapeutic window is narrow, investigating effective diagnostics in low‐resource hospitals is imperative. We used affordable cranial ultrasonography (cUS) and electroencephalography (EEG) to assess how structural abnormalities and impairments in auditory processing are related to neurodevelopmental outcome in East African newborns. Healthy infants (N=15) and infants diagnosed with asphyxia (N=17) were recruited from urban hospitals in Rwanda and Kenya. Structural images of the brain were obtained by cUS through the anterior fontanel. EEG was recorded at rest and during three auditory stimulation tasks. We expect negative health outcomes to be correlated with enlarged lateral ventricle size, thalamus echodensity and the neural strength reflected in the EEG data. We will compare the prognostic value of these measures, after completing “offline” analyses, with clinical assessment of raw data. This study forms a solid foundation for further work in identifying early clinical markers of brain injury and establishing protocols that are viable in the developing world.
Laminin‐coated filaments co‐grafted with Schwann cells and GDNF direct axonal growth following rat thoracic spinal cord injury (729.2)
Authors: Deng Lingxiao , Walker Chandler , Wen Xuejun , Xu Xiao‐Ming
Published: June 2021

Biomaterials are ideal for promoting directional axon growth after spinal cord injury (SCI). We assessed effects of unidirectional polypropylene filaments of varying densities combined with laminin, glial cell line‐derived neurotrophic factor (GDNF) and Schwann cells (SCs) on axon regrowth after T10 spinal hemisection in adult rats. Filaments reduced lesion cavity ~90% compared with non‐filament group and attenuated astrogliosis and inflammation (p < 0.001). Low density filaments promoted more axons (324.4±36.5) and vascular growth (226±49.3) over no filament, middle and high density filament grafting (p < 0.05). Laminin pre‐coated filaments were optimal for axon and vascular regrowth compared with matrigel and hydrogel. To evaluate axon orientation, axons randomly selected at the graft interface were traced by ‘best fit’ lines. The angle between axon lines and filaments parallel to the rostral‐caudal axis of the cord defined orientation (0° = parallel and 90° = perpendicular to the axis). Filament group axon angles were less than the non‐filament group (24.3±7.8° vs. 44.7°±8.3°; p < 0.01). This directional effect was enhanced by co‐grafting SCs and GDNF. Thus, combination of filaments, laminin, SCs and GDNF provided sub‐micron topographic cues for directional axon regrowth revealing new insight for large lesion gap repair and functional recovery after SCI.

Grant Funding Source: NIH grants NS036350, NS052290, NS050243, NS059622, the Daniel Heumann Fund for Spinal Cord Research
Expression and distribution of the scavenger receptor CD36 suggests important roles in the pathophysiology of spinal cord injury (729.3)
Authors: Cordero‐Cabán Kathia , Licero‐Campbell Jennifer , Figueroa Johnny , De León Marino
Published: June 2021

Spinal cord injury (SCI) remains a devastating cause of lifetime medical comorbidities in modern society. Unfortunately, only a few preventive strategies have been shown to prevent the profound pathophysiological mechanisms associated with this condition. In previous studies, our laboratory has shown that a diet enriched in polyunsaturated fatty acids (O3PUFAs), including docosahexaenoic acid (DHA), can reduce neurodegeneration, chronic pain, motor functional deficit, loss of sensitivity, and accelerates autonomic bladder function in a rat model of SCI. CD36 is a multifunctional protein that has been recently implicated as a scavenger receptor and lipid transporter. This membrane protein plays a major role in physiological and pathological inflammation. The purpose of this study was to investigate the expression and roles of CD36 in the context of SCI and dietary O3PUFAs. We hypothesized that trauma to the spinal cord result in a reduction in the protein levels of this protein. Further, we proposed that dietary O3PUFAs normalize the expression of CD36 in the injured spinal cord. Here we determined the protein expression of CD36 after SCI. In particular, we analyzed the temporal and spatial expression of CD36 after contusive injury to the rat spinal cord using immunoblotting and double‐labeling immunofluorescent experiments. Our data demonstrates that acute SCI results in a marked downregulation of CD36. This protein was predominantly observed in both neuronal and glial cells. Importantly, we found that dietary lipids may be implicated in the regulation of this important protein. Altogether, our data supports a potential molecular link between dietary O3PUFAs beneficial effects in the injured spinal cord. This protein may represent an important therapeutic target to reduce the burden of SCI in humans.

Grant Funding Source: Supported by 5P20MD006988 and 2R25GM060507
Bridging the gap: a cell and molecular study of muscle after rotator cuff injury (731.12)
Authors: Gigliotti Deanna , Upadhaya Ritika , Macek Bryce , MacDonald Peter , Leiter Jeff , Anderson Judith
Published: June 2021

Rotator cuff injury is a debilitating condition that surgical intervention often fails to resolve; failure rates range between 30 ‐ 94%. Muscle from 13 patients (4 females, 9 males; 49 ‐ 65 yrs) with rotator cuff injury was studied at a cellular and molecular level to test the notion that it shows signs of atrophy, with or without features related to denervation with the hope of highlighting possible explanations for surgical failures. Satellite cell response to activation via a nitric oxide donor drug called isosorbide dinitrate (ISDN) was also investigated with the idea that satellite cells could be activated during the conservative management phase to promote muscle growth and repair. Supraspinatus and deltoid muscle biopsies taken during arthroscopic rotator cuff repair were analyzed in pairs for histology and showed muscle atrophy in the affected supraspinatus muscle. There were more small diameter fibres (p<0.005) and a smaller mean fibre diameter (17.9 ± 0.3μm) in supraspinatus vs. control deltoid muscle (21.8 ± 0.3μm & p<0.003). Alpha‐bungarotoxin staining of acetylcholine receptors and western blotting for their gamma and epsilon subunits showed tendencies (p=0.13) to a linear arrangement and increased gamma to epsilon ratio respectively; both are signs of denervation. Response to ISDN showed more mitotically active Pax7+ satellite cells in supraspinatus muscle cultured with ISDN than in untreated supraspinatus or in the deltoid (p<0.03). Results are consistent with the notion that supraspinatus muscle of the injured rotator cuff is atrophic with possible denervation; together these features may increase susceptibility to muscle injury and surgical failure. Results also suggest the potential for the using ISDN to promote muscle growth and increase surgical success.

Grant Funding Source: Supported by Canadian Orthopedic Foundation, Pan Am Clinic Foundation & Department of Surgery
Effect of exercise on limb muscle fiber architecture in a Pompe mouse model (731.4)
Authors: Daly Shane , Case Laura , Pastva Amy , Kishnani Priya , Koeberl Dwight , Taylor Andrea
Published: June 2021

Pompe disease, a glycogen storage disease causing muscle weakness, is treated by enzyme replacement (ERT). As part of a larger study of exercise effects, we studied the impact of exercise on limb muscle fiber architecture using a mouse knockout model (GAA‐KO) on ERT via gene therapy. Thirty‐seven mice were assigned to 1 of 3 exercise groups: high treadmill (HT), low treadmill (LT), voluntary wheel (WL) vs. sedentary control (SED) and sacrificed after eight weeks. Muscle weight, fiber length (Lf) and in situ sarcomere length were measured and physiologic cross‐sectional area computed for tibialis anterior (TA), gastrocnemius (GA) and soleus (SOL). Initial results: The nonparametric Kruskal‐Wallis revealed significant group differences in Lf for TA (p=.030) and GA (p=.049). Post‐hoc Mann‐Whitney U tests showed significantly longer TA fibers in WL and HT vs. LT (p=.034) and a trend toward longer fibers in WL vs. SED (p=.059). GA fibers were significantly shorter (p=.034) in HT and WL vs. LT. We observed a trend toward shorter GA fibers in HT vs. all other groups and in WL vs. SED (p=.083). Lengthened TA fibers and correspondingly shorter GA fibers may indicate dorsiflexion weakness, as in human infantile onset Pompe disease on ERT, supporting prior concerns about potential negative effect of more strenuous exercise regimens. Further data analysis will continue to explore effects of exercise on muscle strength and function.

Grant Funding Source: Supported by R24 HD050837‐01 and NSF BCS 0962677
Blockade of receptor activator of nuclear factor‐κB signaling greatly improves SERCA activity and contractility in atrophic and dystrophic skeletal muscles (731.7)
Authors: Dumont Nicolas , Dufresne Sebastien , Dufresne Sebastien , Boulanger‐Piette Antoine , Lavergne liane , Bouchard Patrice , Penninger Josef M. , Frenette Jrme
Published: June 2021

Receptor‐activator of nuclear factor‐κB (RANK), its ligand RANKL and the soluble decoy receptor osteoprotegerin (OPG) are members of the tumor necrosis factor (TNF) superfamily that control osteoclast differentiation, bone remodelling and osteoporosis. Since bone and skeletal muscle physiopathology occurs synchronously, we tested whether RANK/RANKL pathway is involved in muscle dysfunction following different muscle wasting conditions. Contractile properties of EDL muscles were obtained following 14d of denervation in mice specifically deficient in RANK skeletal muscle or in dystrophic mdx mice treated daily for 10d with OPG [0.3mg/kg], the natural inhibitor of RANKL. The activity and expression of sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) were determined in denervated or dystrophic muscles. RANK muscle deletion did not prevent atrophy but greatly reduced the losses of force production and SERCA activity in denervated EDL muscles. Force production of EDL muscles also increased by 140% in OPG‐treated relative to PBS‐treated mdx mice while the treatment with OPG prevented the loss of the fast twitch SERCA1 protein content in dystrophic mice. Together, these data indicate that the deletion or blockage of RANK/RANKL pathway has inotropic effects and increases SERCA activity and expression in fast‐twitch skeletal muscles opening promising new avenues for several muscle diseases.
An increase in microRNA‐23 prevents muscle wasting in mice with chronic kidney disease in resistance exercise (731.8)
Authors: Wang Xiaonan
Published: June 2021

In our previous studies we found that exercise partially prevents chronic kidney disease (CKD)‐induced muscle atrophy. We also saw that microRNA‐23 (miR‐23) is decreased in muscles of CKD mice. In silico analysis suggested that miR‐23 targets several proteins associated with muscle atrophy. In this study, we evaluated whether exercise restores the level of miR‐23 in CKD mice. CKD was induced in 20‐25g mice by 5/6th nephrectomy. Muscle overloading, a resistance exercise model, was produced in both control and CKD mice by removing the gastrocnemius and soleus muscles from both hindlimbs. Overexpression of miR‐23 in cultured muscle cells inhibits the reporter luciferase activities of PTEN and YY1, suggesting that miR‐23 attenuates the expression of these predicted targets. Exercise significantly increased the level of miR‐23 in muscle of CKD mice vs. unexercised CKD mice. Consistent with this, exercise decreased PTEN protein and increased consequent Akt phosphorylation (2.3‐fold, P<0.001) which would limit muscle atrophy. In CKD mice, exercise increased MyoD, myogenin and eMyHC, proteins that are linked to the myogenic transcription factor, YY1. Conclusions: Resistance exercise can increase miR‐23 which leads to a suppression of multiple atrophy‐related target proteins that may contribute to the muscle‐sparing effects of exercise in CKD.

Grant Funding Source: NIH AR060268 (XHW)
Low intensity laser irradiation in myogenic precursor cells: an in vitro study (732.4)
Authors: Amaral André , de Araújo Heloisa , Parizotto Nivaldo , Salvini Tânia
Published: June 2021

The aim of this study was to analyze the influence of low intensity laser irradiation at wavelengths (λ) of 670, 685 and 830 nm on the mitochondrial activity and gene expression profile of myogenic precursor cells (MPCs) in vitro. Primary rat MPCs were sown in multiwell dishes of 96 and 6 wells. Six experimental groups were set up: a non‐irradiated control group and five groups ranged by differing doses of radiation (0.3; 0.6; 0.9; 1.2 and 1.5 J/cm2). After 24 hours of irradiation, the mitochondrial activity was quantified using the colorimetric technique MTT‐formazan and established in terms of percentage in relation to the control. The gene expression profile was established through the quantitative analysis of the related isoforms IIa and IIx of the myosin heavy chain (MHC) by RT‐PCR five days after irradiation. The results showed that irradiation at λ of 670, 685 and 830 nm promoted respective increase of 56.8, 70.6 and 84.8 % on mitochondrial activity. These values were achieved with the 1.2, 0.6 and 0.3 J/cm2. As far as concerns MHC isoforms expression, this showed that there was a significant decrease in the IIa isoform (685 nm) and an increase in the IIx isoform (685 and 830 nm) in comparison to the non‐irradiated cells. Laser radiation was characterized as being a wavelength and dose‐dependent effect on the intensity of mitochondrial activity and gene expression profile among MPCs in vitro.

Grant Funding Source: Supported by: Fundação de Amparo à Pesquisa do Estado se São Paulo ‐ FAPESP
Interleukin‐6 and cyclooxygenase‐2 mediate myofibroblast‐induced polarization of alternatively activated macrophages (734.9)
Authors: Fernando Maria , McKay Derek
Published: June 2021

Alternatively activated macrophages (AAMΦs) are a macrophage subset induced primarily by the Th2 cytokines IL‐4 and IL‐13. They are associated with anti‐inflammatory functions, such as wound healing and tissue repair and are also protective in animal models of diseases, such as inflammatory bowel disease and multiple sclerosis. In this study we showed that myofibroblasts, also associated with wound healing, may have a putative in vivo role in the induction and maintenance of AAMΦs. Macrophages and myofibroblasts (MFbs) were derived from murine bone marrow and dermal tissue (wild‐type and IL‐6‐/‐), respectively. MΦ were exposed to IL‐4+IL‐13 conditioned medium (CM) from control MFbs or MFbs treated with a non‐selective cyclooxygenase (COX)‐1 and ‐2 inhibitor (indomethacin) or a selective COX‐1 (SC‐560) or COX‐2 inhibitor (NS‐398). Exposure to control MFb CM enhanced markers of AAMΦ polarization and reduced LPS‐evoked nitric oxide release. These effects were partially reduced using MFbs from IL‐6‐/‐ mice, suggesting a role for IL‐6 in this process. CM from indomethacin‐ and NS‐398‐treated MFbs failed to enhance an AAMΦ phenotype ‐ SC‐560 did not have this same effect. Inhibition of EP4 on MΦ also eliminated the effect of MFb CM, implicating PGE2. These data indicate a role for IL‐6 and PGE2, two molecules commonly linked with pro‐inflammatory functions, in the differentiation of anti‐inflammatory AAMΦs.

Grant Funding Source: CCFC, CIHR, CDHF, AIHS
Identification of novel aldosterone targets in mouse kidney cortical collecting duct cells (744.1)
Authors: Gumz Michelle , Richards Jacob , Solocinski Kristen
Published: June 2021

The kidney is critical for the regulation of blood pressure by maintaining sodium and fluid homeostasis. The hormone aldosterone is critical for the regulation of these processes. Aldosterone acts on principal cells of the renal collecting duct to increase sodium reabsorption by increasing transcription of the epithelial sodium channel and SGK1; these effects are mediated by the mineralocorticoid receptor (MR). Recent work by others identified two novel factors that may play a role in sodium reabsorption in the kidney. FK506 binding protein 5 (FKBP5), a member of the immunophilin family, is a co‐chaperone for MR, and causes increased translocation of MR into the nucleus. Growth arrest‐specific 5 (Gas5) is a long noncoding RNA (lncRNA) that acts as a decoy to the hormone response element (HRE) of MR. Gas5 may compete for binding of MR to the HREs of aldosterone target genes. However, it is unknown if FKBP5 or Gas5 are aldosterone targets. Thus we treated mouse kidney collecting duct cells (mpkCCDc14), with aldosterone for 24 hours. Aldosterone treatment led to increases in FKBP5 and Gas5 RNA expression. These data demonstrate that FKBP5 and Gas5 are potential aldosterone targets in kidney cortical collecting duct cells. This may be the first report demonstrating aldosterone‐mediated regulation of a lncRNA.

Grant Funding Source: Supported by NIH, ASN and AHA
Neuroprotective effect of estrogen on hypoglycemic injury in hypothalamic mouse cells (767.6)
Authors: Alicea Eilliut , Chakraborty Tandra , Cohen Joshua , Yohanan Darien , Chakraborty Sanjoy
Published: June 2021

Glucose is the main metabolic fuel of the brain. Recurrent episodes of hypoglycemia can lead to seizures, coma, and even death. The hypothalamus; a region involved in feeding and energy balance is most vulnerable to this injury. In the present study we report neuroprotection of estrogen on hypoglycemic injury in a hypothalamic cell line. The cell viability, toxicity and proliferation were determined using MTT, lactate dehydrogenase (LDH) assay, and cell count. There was a significant increase in the cell count and increase in mitochondrial function in the presence of estrogen (p<0.0001) when exposed to hypoglycemia within 24 hours. The attenuation of the death pathways was only seen within the first 24 hours and gradually decreased with time. Percent cytotoxicity also increased in the absence of glucose and decreased in the presence of estrogen within 24 hours. Two pathways seemed viable in mediating estrogen action, the phospho‐AKT pathway, and a pathway involving caveolin‐1 (CAV‐1). In order to determine estrogen mode of action, PRAS 40 was analyzed via ELISA assay, which showed no change when exposed to hypoglycemic shock. This suggested that the phospho‐AKT pathway was not involved in estrogen mode of action. Our current efforts are focused on determining whether estrogen neuroprotection is mediated through CAV‐1 and re‐testing the phospho‐AKT pathway via western blot analysis.
Genetic variation affecting temporal patterns of gene expression and disease in human brain (776.1)
Authors: Fraser Hunter , Martin Trevor
Published: June 2021

It is widely recognized that noncoding regions of the human genome, including those controlling the cis‐regulation of gene expression, harbor most disease‐associated genetic variants and most regions under selective constraint. Although the expression of many genes is a dynamic process that changes during an organism’s lifetime, most studies mapping genetic variants affecting gene expression have either ignored the variation in ages among individuals, or treated it as a confounding variable. Here we introduce a method for mapping genetic variants that affect the developmental timing of gene expression, which we call temporal expression quantitative trait loci (teQTL). Applying this approach to human prefrontal cortex, we found several thousand genes associated with teQTLs. These teQTLs are distinct from variants affecting expression levels throughout life, and are often associated with distal transcriptional enhancers or age‐related changes in DNA methylation. Moreover they coincide with dozens of SNPs previously implicated in neurological traits such as Alzheimer’s disease, Parkinson’s disease, and educational attainment. These results suggest that teQTLs are a widespread source of variation in human neurological phenotypes.

Grant Funding Source: Supported by the Pew Foundation
Clocking regional chemotherapy of liver metastases (780.2)
Authors: Turebaev D. , Urazova S.
Published: June 2021

Clocking regional chemotherapy ‐ is an effective treatment for neoplastic lesions of the liver at a new modern level.

Clocking regional chemotherapy was held in 58 patients (25 men and 33 women) with liver metastases of primary source different localization. Clocking method of regional chemotherapy is a method of improved chemotherapy, where drugs are introduced by the digital dosers, taking into account the pharmacodynamics of drugs and simultaneous administration of supporting drugs. The new modern digital infusion pumps and micropumps have been used, that allowed to keep a constant dose administration of drugs, their dosage requirement directly by the patient.

The results of clinical observations have shown that with adequate controlled introduction of drugs increases their therapeutic effect.

Improved schemes for using domestic Erbisol more clearly disclose its hepatoprotective properties and reparative and selectivelyimmunomodulatory properties ‐ stimulator of T‐ lymphocytes and NK cells.

One of the important achievements of the proposed method the clocking regional chemotherapy is an active participation of the patient in the conducting therapy. It is controlled analgesia during manipulation and treatment ‐ the amount of pain medication was reduced by 35‐50%, further patient‐controlled administration of antiemetic drugs ‐ the volume was reduced by 40‐65%.
Biocompatible evaluation of fluorescent nanodiamonds in neurons in vitro (780.5)
Authors: Kao Jun‐Wei , Huang Yung‐An , Chiang Ming‐Han , Huang Hou‐Syun , Liu Kuang‐Kai , Chao Jui‐I , Hwang Eric
Published: June 2021

In recent years, nanodiamond has emerged as a promising biomaterial for diagnostic and therapeutic applications. Fluorescent nanodiamond (FND) containing nitrogen‐vacancy centers is a new addition to the nanodiamond family. The ﬂuorophore produced by nitrogen‐vacancy made FND extremely photostable. Furthermore, the high refractive index makes FNDs visible under light microscopy. In previous studies, FND has been reported to have outstanding biocompatibility and high uptake rate in various cell types. However, the application of FND on neurons or nervous tissue remains elusive. In order to study the compatibility of FND on neurons, the viability and morphological alteration of neurons after treated with FND were examined. Dissociated primary neurons from both central nervous system (CNS) and peripheral nervous system (PNS) were cultured in the presence of FND and neither exhibited a reduction in viability. However, we did observe a FND dosage‐dependent decrease in neurite length in both CNS and PNS neurons. Time‐lapse live cell imaging suggested that the reduction of neurite length was due to the spatial hindrance of FND on advancing neuronal growth cone. Additionally, neuronal uptake of FNDs was confirmed using confocal microscopy and flow cytometry. These results indicated that FND exhibited low neuronal toxicity and can be readily taken into neurons. However, high dosage of FND may interfere with neurite elongation and should be taken into consideration when applications involve actively growing neurites (e.g. axon regeneration).

Grant Funding Source: Supported by National Science Council of Taiwan
NSAIDs inhibit cell migration and calpain expression in rat duodenum (784.1)
Authors: Silver Kristopher , Bawa Bhupinder , Lillich James
Published: June 2021

Nonsteroidal anti‐inflammatory drugs (NSAIDs) are one of the most commonly used drugs for the alleviation of pain and inflammation. Unfortunately, daily use of NSAIDs is associated with significant adverse side effects, including erosion and ulceration of the gastrointestinal (GI) tract. Previous studies have shown that NSAIDs inhibit migration of IEC‐6 cells (a small intestinal epithelial cell line) by inhibiting calpain activity and membrane expression. The current study was undertaken to determine if NSAIDs affect calpain expression and cell migration in vivo. Rats were treated with indomethacin or NS‐398 for 72 h and injected with bromodeoxyuridine (BrdU) at 6, 12, 18, or 24 h prior to tissue harvest. Sections of duodenum were harvested for RNA and protein extraction as well as for histomorphometric analysis of crypt and villous length and anti‐BrdU immunohistochemistry to estimate cell migration. Our results show that there was stunting of villi and a reduction in cell migration in rat duodenum by NSAID treatment. In addition, protein expression for calpains 1, 2, and 8 was significantly reduced in duodenal mucosa by NSAID treatment despite enhanced expression of calpain 8 mRNA. These results suggest that reductions in calpain expression in vivo are associated with deficits in cell migration caused by NSAIDs, and that calpains may be clinically important targets of NSAID‐induced GI toxicity.

Grant Funding Source: Supported by the National Institute of Diabetes and Digestive and Kidney Diseases (1R15DK091791)
In vivo exploration of the significance of N‐glycolylneuraminic acid exclusion from the vertebrate brain (787.3)
Authors: Naito‐Matsui Yuko , Davies Leela , Takematsu Hiromu , Chou Hsun‐Hua , Tangvoranuntakul Pam , Carlin Aaron , Verhagen Andrea , Gagneux Pascal , Varki Ajit
Published: June 2021

Sialic acids are acidic sugars often occupying the non‐reducing terminus of glycans. In mammals, the major sialic acids are N‐acetylneuraminic acid (Neu5Ac) and N‐glycolylneuraminic acid (Neu5Gc). The nucleotide donor CMP‐Neu5Gc is biosynthesized from CMP‐Neu5Ac by CMP‐Neu5Ac hydroxylase (Cmah). Regulation of Cmah expression results in markedly varying ratios of Neu5Ac and Neu5Gc among different tissues in different animal species. However, the brain shows extremely low Neu5Gc expression (<3%) in all species tested to date. We used Neu5Gc‐overexpressing Cmah transgenic mice to ask if brain expression of Neu5Gc negatively affects development and/or function. In initial attempts using CMV or NSE promoters, no live mice overexpressing Neu5Gc were obtained, suggesting toxicity. Next, we applied a Cre‐loxP approach for controlled transgene‐derived Cmah expression by Cre recombinase. Interestingly, forced Cmah expression did not show lethality or gross developmental defects with several different Cre mouse lines. The transgenic mice showed remarkably high and widely distributed Neu5Gc expression in the whole brain. Thus, the embryonic lethality seen initially is due to some other effect of Neu5Gc overexpression. This mouse model will provide new insights in understanding unique features of sialic acids in the brain. We are currently studying sialoglycan structures and phenotypes in such mice.

Grant Funding Source: NIH R01 CA38701
Functional characterization of the N‐glycans in lysosomal alpha‐mannosidase revealed a different sorting mechanism in the brain compared to peripheral organs (787.4)
Authors: Greiner‐Tollersrud Ole
Published: June 2021

Functional characterization of the N‐glycans in lysosomal alpha‐mannosidase revealed a different sorting mechanism in the brain compared to peripheral organs. Ole K. Greiner‐Tollersrud, Christophe Flahaut, Jean C. Michalski, G. Evjen, Hilde M.F. Stensland, Karl E. Eilertsen, G. Hansen: VBRG, IMB, University of Tromso, 9037 Tromso, Norway Lysosomal alpha‐mannosidase (LAMAN) is a homodimer of 250 kDa, which is sorted to the lysosomes via the mannose 6‐P dependent pathway. N‐glycans linked to specific sites or regions of the folded LAMAN surface exerted multiple roles: 1) ER to Golgi transport of nascent LAMAN which requires an N‐glycan linked to a small region including positions 310 and 367; 2) Trafficking from Golgi to lysosomes via the mannose 6‐P dependent pathway which requires an N‐glycan linked to an accessible surface region of 40 Å in diameter, including positions 310, 367, 766 and 930; 3) Conformational stability in the lysosomes exerted by a high mannose structure linked to N497 within the dimer cleft; 4) Protection from lysosomal proteolyses by critically positioned N‐glycosylation sites in some species The surface region to which mannose‐phosphorylated glycans were attached, the “footstep” of GlcNAc 1‐phosphotransferase, included only one of the two LAMAN monomers, since to each of the N310, N367, N766 and N930‐sites there were linked two glycan types: A) GlcNAc2Man5‐7‐structures which are likely lysosomally processed forms of mannose 6‐phosphorylated glycans and B) Fucosylated paucimannosidic/bisected glycans which are processed forms of complex type glycans. The N‐glycans linked to the accessible N‐glycosylation sites outside this “footstep” were processed to endo H‐resistant complex glycans in bovine liver, kidney and lung. In bovine brain, however, these N‐glycans remained mainly endo H‐sensitive, and likely processed into hybrid type structures. Thus, brain LAMAN may be subjected to a unique trafficking diversion during transit through the Golgi complex.
Antioxidant activity of flavanoids on basis of vegetation raw material of Kazakhstan (794.2)
Authors: Zeinuldina Aizhan , Gabbasova Anar , Tuleuov Borash , Abduldayeva Aigul
Published: June 2021

The most important of flavanoids in metabolism in vegetable cell is connected with its ability to make up tough chelate complexes with various ions of metals. Also the ability to interact with free radicals, to participate in electrons’ transport, to connect with various enzymes, changing their activity. Flavanoids are able to penetrate into cell membranes where the flavanoids destroy the primary and by‐productive of lipids peroxidation (PLP). As we know pathological state in human’s body in that or another degree is closely connected with the processes PLP. We have made research of the comparative study of individual compounds of routine, quercertin’s influence and total medicines on the basis of hawthorn blood‐red, Russian thistle and saw‐wort upon the dynamics of lipids peroxidation under strict gycopsy at laboratory rats. About the PLP’s condition we have judged by the maintenance of malonic dialdehyde (MDA). Antioxidant activity we have valued by changing catalase activity in serum blood but antioxidant effect by inhibition peroxidation of liposomes at presence exogenous substances. It should be in mind that changes of catalase activity adequate antioxidant effect of having researched medicines. More marked antioxidant activity was expressive quercetin which can be explained by two factors: by the same doze of medicines, the less molar mass quercetin rises the concentration of substances.
Pharmacological evaluation of novel furan‐2‐carboxamide derivatives as antihyperlipidemic agents in triton‐induced rats (794.4)
Authors: Al‐Qirim Tareq , Shattat Ghassan , Abu‐Skeika Ghassan , Alhusaini Nadeem
Published: June 2021

A new series of Furan‐2‐carboxamide derivatives of anthraquinone (compounds N1, N3, N5) were synthesized and tested in vivo using Triton WR‐1339‐induced hyperlipidemic rat as an experimental model for their hypolipidemic activity. The tested animals were divided into six groups: control, hyperlipidemic, N1, N3, N5 and bezafibrate. The designed target derivatives, compounds N1, N3 were inactive. Compound N5 has shown a significant reduction in triglyceride level in rats by 66.5% after 18 h in comparison to the hyperlipidemic group. Furthermore, high‐density lipoprotein‐cholesterol levels were remarkably increased in compound N5 (p < 0.001) after 18 h in comparison to the hyperlipidemia group and also compound N5 significantly reduced LDL‐C level (p< 0.001) after 18 h in comparison to the hyperlipidemia group. Furthermore, the total cholesterol level was reduced by 85% after 18 h in compound N5 compared to HLG‐treated rats. The present study demonstrated new properties of novel series of furan‐2‐carboxamides N5 as a potent lipid‐lowering agent. It is therefore reasonable to assume that this compound may have a promising potential in the treatment of hyperlipidemia and coronary heart diseases.

Grant Funding Source: This research is supported by Al‐Zaytoonah University of Jordan
A weak PAR2 partial agonist that can block PAR2 signaling initiated via protease and/or potent peptidomimetics (794.5)
Authors: Boitano Scott , Flynn Andrea , Hoffman Justin , Asiedu Marina , Zhang Zhenyu , Sherwood Cara , DeFea Kathryn , Vagner Josef , Price Theodore
Published: June 2021

Protease‐activated receptor‐2 (PAR2) is a GPCR activated by proteolytic cleavage of the extracellular NH2‐terminus resulting in a newly exposed peptide sequence or “tethered ligand.” Potent and efficacious peptidomimetic agonists have been developed to PAR2, however, similar antagonists have been lacking. We have modified the peptidomimetic agonist, 2‐furoyl‐LIGRLO‐NH2, substituting LIG2‐4 with an azabicycloalkane scaffold and O6 with a Tyr. At 10 ‐ 50 M, C391 blocks protease and/or peptidomimetic induced, PAR2 dependent, Ca2+ and MAPK signaling and, ‐arrestin recruitment. In a PAR2‐dependent behavioral mouse model, C391 dose‐dependently blocked (75 g max effect) mast cell degranulation‐induced thermal hyperalgesia. We transformed C391 into a synthetic tethered ligand (C391‐PEG3‐Hdc) in an effort to increase PAR2 affinity and efficacy. C391‐PEG3‐Hdc performed as a partial agonist across PAR2 assays. We conclude that C391 is a weak partial agonist with low M PAR2 affinity that can act as an antagonist when applied at appropriate concentrations. This represents a pharmacological mechanism of action for PAR2 that can be fruitful for the development of therapeutics.

Grant Funding Source: NIH‐NS073664
High fat diet induces aberrant COX‐2 gene expression through demethylation of a transcriptional enhancer at a 5’ upstream region of the gene (800.7)
Authors: Hernandez Saavedra Diego , Zhou Dan , Liu Debra , Chen Hong , Pan Yuan‐Xiang
Published: June 2021

Cyclooxygenase‐2 (COX‐2) produces prostaglandins that participate in multiple physiological and pathological processes, including the activation of inflammatory responses. Induction of COX‐2 gene expression has been reported to be correlated with DNA hypomethylation. This study characterized a novel enhancer located at an upstream region on the COX‐2 gene and associated alterations of the DNA methylation patterns by high fat diet. Male Sprague‐Dawley rats received a high fat diet at different life stages, including maternal (HF/C), post‐weaning (C/HF), and lifelong (HF/HF). Liver was collected for analysis at 12 wks of age. Results showed that the high fat diet induced the expression of COX‐2 mRNA in all three high fat groups when compared to the control group (C/C). Genome‐wide methylated DNA immunoprecipitation (MeDIP) showed that DNA hypomethylation occurred at an upstream region of distal promoter of COX‐2 gene in all three high fat groups. Site‐specific hypomethylation of CpG in this region was further analyzed by bisulfite sequencing. Luciferase reporter assay demonstrated an enhancer activity of this particular region. Thus, it is likely that exposure to a high fat diet at various stages of life induces COX‐2 expression through the reduction of DNA methylation at an upstream enhancer region of the gene.
Altered expression of neuroplasticity related genes is associated with pathophysiology of human depression: expression analysis in postmortem human brain of depressed suicide victims (803.13)
Authors: Fuchsova Beata , Alvarez Julia Anabel , Rizavi Hooriyah , Frasch Alberto Carlos , Pandey Ghanshyam
Published: June 2021

Disruptions in normal plasticity mechanisms have been shown to play a part in the etiology of depression. Likewise, chronic stress, a well‐established risk factor for depression, affects plasticity: reduces dendritic spines, synapses, as well as the length and complexity of dendrites in the hippocampus and prefrontal cortex (PFC).

GPM6A, a neuronally expressed member of the proteolipid protein family, is a key modulator for neurite and filopodium outgrowth, and synapse formation. Previously, we identified GPM6A gene as a stress‐responsive gene downregulated in the hippocampus in animal models of chronic stress. This effect is reversed by antidepressants. Signaling pathways that mediate GPM6A neuroplastic function and its relevance to human depression are unknown.

Here we used qPCR to find out whether GPM6A is relevant to the pathogenesis of human depression. For the first time, we show that GPM6A is transcriptionally downregulated in the postmortem hippocampal tissue of suicide subjects with depression. Using coimmunoprecipitation of GPM6A followed by mass spectrometry we identify calcium/calmodulin dependent protein kinase II α, G protein‐coupled receptor kinase‐interacting protein 1, and coronin 1A to form complexes with GPM6A. We show that also mRNA level of these genes and the coordinated transcriptional response is altered in postmortem brain of depressed suicide subjects.

Grant Funding Source: ANPCyT
Expression of microRNAs associated with apoptosis in neurospheres and adhered cells in glioblastoma cell line culture treated with ionizing radiation and temozolomida (803.14)
Authors: Rodrigues Andressa , Trevisan Felipe , Peria Fernanda , Carlotti Junior Carlos , Tirapelli Daniela
Published: June 2021

Objectives: Evaluating the effect of ionizing radiation and temozolomide, alone or associated, on the expression of the miRNAs miR‐15, miR‐16 and miR‐21 in neurospheres and adhered cells in glioblastoma cell line (U343‐MG).

Methods: The trypan blue was used to examine cell viability before and after treatment, and real time PCR to analyze the expression of microRNAs in time 0h (immediately after treatments), and 48h after the exposure to treatments.

Results: The analysis of cell viability showed no statistically significant difference among the studied groups. The oncogene miR‐21 demonstrated a significantly higher expression in neurospheres in group treated with ionizing radiation (both 0h and 48h). The tumor suppressors microRNAs miR‐15 and miR‐16 had no significant expression among cell types and treatments modalities submissions. However, miR‐15 and miR‐16 had a considerably different expression between the analyzed periods: miR‐15 in the time 0h was highly expressed in adherent cells treated with ionizing radiation and temozolomide; and, during the 48 hour analysis, miR‐15 was more expressed in neurospheres with the same type of treatment. The same expression profile happened with the miR‐16, however being more expressed in the group treated with ionizing radiation.

Conclusions: MicroRNAs studied in glioblastoma cell line culture demonstrated differentially expressions when compared neurospheres and adhered cells and different treatments. MiR‐21 expression was significantly different between cell types (neurospheres and adhered cells), remaining more expressed in neurospheres treated with ionizing radiation.