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1

Ferraris, Christopher M., Rebecca Dunayev, Nour Kanaan, et al. "93 Acceptability and Usability of Tablet-Based Neuropsychological Tests among South African and Ugandan Adolescents With and Without HIV." Journal of the International Neuropsychological Society 29, s1 (2023): 495–96. http://dx.doi.org/10.1017/s1355617723006410.

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Objective:Neuropsychological (NP) tests are increasingly computerized, which automates testing, scoring, and administration. These innovations are well-suited for use in resource-limited settings, such as low- to middle- income countries (LMICs), which often lack specialized testing resources (e.g., trained staff, forms, norms, equipment). Despite this, there is a dearth of research on their acceptability and usability which could affect performance, particularly in LMICs with varying levels of access to computer technology. NeuroScreen is a tablet-based battery of tests assessing learning, memory, working memory, processing speed, executive functions, and motor speed. This study evaluated the acceptability and usability of NeuroScreen among two groups of LMIC adolescents with and without HIV from Cape Town, South Africa and Kampala, Uganda.Participants and Methods:Adolescents in Cape Town (n=131) and Kampala (n=80) completed NeuroScreen and questions about their use and ownership of, as well as comfort with computer technology and their experiences completing NeuroScreen. Participants rated their technology use -comfort with and ease-of-use of computers, tablets, smartphones, and NeuroScreen on a Likert-type scale: (1) Very Easy/Very Comfortable to (6) Very Difficult/Very Uncomfortable. For analyses, responses of Somewhat Easy/Comfortable to Very Easy/Comfortable were collapsed to codify comfort and ease. Descriptive statistics assessed technology use and experiences of using the NeuroScreen tool. A qualitative question asked how participants would feel receiving NeuroScreen routinely in the future; responses were coded as positive, negative, or neutral (e.g., “I would enjoy it”). Chi-squares assessed for group differences.Results:South African adolescents were 15.42 years on average, 50.3% male, and 49% were HIV-positive. Ugandan adolescents were 15.64 years on average, 50.6% male, and 54% HIVpositive. South African participants were more likely than Ugandan participants to have ever used a computer (71% vs. 49%; p<.005), or tablet (58% vs. 40%; p<.05), whereas smartphone use was similar (94% vs 87%). South African participants reported higher rates of comfort using a computer (86% vs. 46%; p<.001) and smartphone (96% vs. 88%; p<.05) compared to Ugandan participants. Ugandan adolescents rated using NeuroScreen as easier than South African adolescents (96% vs. 87%; p<.05).). Regarding within-sample differences by HIV status, Ugandan participants with HIV were less likely to have used a computer than participants without HIV (70% vs. 57%; p<.05, respectively).The Finger Tapping test was rated as the easiest by both South African (73%) and Ugandan (64%) participants. Trail Making was rated as the most difficult test among Ugandan participants (37%); 75% of South African participants reported no tasks as difficult followed by Finger Tapping as most difficult (8%). When asked about completing NeuroScreen at routine doctor’s visits, most South Africans (85%) and Ugandans (72%) responded positively.Conclusions:This study found that even with low prior tablet use and varying levels of comfort in using technology, South African and Ugandan adolescents rated NeuroScreen with high acceptability and usability. These data suggest that scaling up NeuroScreen in LMICs, where technology use might be limited, may be appropriate for adolescent populations. Further research should examine prior experience and comfort with tablets as predictors NeuroScreen test performance.
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2

Steenkamp, Nina S., Hetta-Mari Gouse, Rhiannon Changuion, et al. "32 Influence of Prior Experience with Computer-Based Technology on Tablet-Based Neurocognitive Test Performance: Data from a sample of cognitively impaired South African older adults." Journal of the International Neuropsychological Society 29, s1 (2023): 713–14. http://dx.doi.org/10.1017/s1355617723008901.

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Objective:The global prevalence of persons living with dementia will soon exceed 50 million. Most of these individuals reside in low- and middle-income countries (LMICs). In South Africa, one such LMIC, the physician-to-patient ratio of 9:10 000 severely limits the capacity of clinicians to screen, assess, diagnose, and treat dementias. One way to address this limitation is by using mobile health (mHealth) platforms to scale-up neurocognitive testing. In this paper, we describe one such platform, a brief tablet-based cognitive assessment tool (NeuroScreen) that can be administered by lay health-providers. It may help identify patients with cognitive impairment (related, for instance, to dementia) and thereby improve clinical care and outcomes. However, there is a lack of data regarding (a) the acceptability of this novel technology for delivery of neurocognitive assessments in LMIC-resident older adults, and (b) the influence of technology-use experience on NeuroScreen performance of LMIC-resident older adults. This study aimed to fill that knowledge gap, using a sample of cognitively impaired South African older adults.Participants and Methods:Participants were 60 older adults (63.33% female; 91.67% right-handed; age M = 68.90 years, SD = 9.42, range = 50-83), all recruited from geriatric and memory clinics in Cape Town, South Africa. In a single 1-hour session, they completed the entire NeuroScreen battery (Trail Making, Number Speed, Finger Tapping, Visual Discrimination, Number Span Forward, Number Span Backward, List Learning, List Recall) as well as a study-specific questionnaire assessing acceptability of NeuroScreen use and overall experience and comfort with computer-based technology. We summed across 11 questionnaire items to derive a single variable capturing technology-use experience, with higher scores indicating more experience.Results:Almost all participants (93.33%) indicated that NeuroScreen was easy to use. A similar number (90.00%) indicated they would be comfortable completing NeuroScreen at routine doctor's visits. Only 6.67% reported feeling uncomfortable using a tablet, despite about three-quarters (76.67%) reporting never having used a tablet with a touchscreen before. Almost one in five participants (18.33%) reported owning a computer, 10.00% a tablet, and 70.00% a smartphone. Correlations between test performance and technology-use experience were statistically significant (or strongly tended toward significance) for most NeuroScreen subtests that assessed higherorder cognitive functioning and that required the participant to manipulate the tablet themselves: Trail Making 2 (a measure of cognitive switching ability), r = .24, p = .05; Visual Discrimination A (complex processing speed [number-symbol matching]), r = .38, p = .002; Visual Discrimination B (pattern recognition), r = .37, p = .004; Number Speed (simple information processing speed), r = .36, p = .004. For the most part, there were no such significant associations when the NeuroScreen subtest required only verbal input from the participant (i.e., on the list learning and number span tasks).Conclusions:NeuroScreen, a tablet-based neurocognitive screening tool, appears feasible for use among older South Africans, even if they are cognitively impaired and have limited technological familiarity. However, test performance might be influenced by amount of technology-use experience; clinicians using the battery must consider this in their interpretations.
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3

CE, Oldham, Wooten CJ, and Lopez D. "Thyroid Hormone Enhances Neurite Outgrowth in Neuroscreen 1 Cells." International Journal of Biomedical Investigation 1, no. 1 (2018): 1–11. http://dx.doi.org/10.31531/2581-4745.1000104.

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4

Dayami, Lopez. "Thyroid Hormone Enhances Neurite Outgrowth in Neuroscreen 1 Cells." International Journal of Biomedical Investigation 1, no. 1 (2018): 1–11. https://doi.org/10.31531/2581-4745.1000104.

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<em>Objectives: Alzheimer&rsquo;s disease (AD) is a neurodegenerative disorder that affects millions of individuals. Moreover, hypothyroidism has been identified as one of the risk factors that may contribute to the development of AD. Here, we investigated whether there was a correlation among expression levels of proteins involved in the formation of AD lesions, neurite outgrowth, and thyroid hormone levels. </em> <em>Methods: Cells were grown in media supplemented with different levels of 3,5,3&rsquo;-triiodothyronine (T3) and then processed for neurite outgrowth and to prepare RNA samples.&nbsp; RNA samples were analysed using quantitative real-time PCR.&nbsp; Protein levels were measured using in cell-Western blotting analysis.</em> <em>Results: By using neurite outgrowth studies, it was demonstrated that T3 treatment enhanced neurite outgrowth in NS-1 cells in a time- and dose-dependent manner. Quantitative real-time PCR studies further confirmed that NS-1 cells expressed substantial levels of TR&alpha; and significantly less TR&beta;, either of which could be responsible for the T3-dependent effects on neurite outgrowth.&nbsp; Although the overall tau protein expression was not affected in response to T3 treatment, the splicing of tau exon 10 was impacted in the direction of producing more tau molecules that excluded the exon (tau 3R).&nbsp; </em> <em>Conclusion: The results of this study are critical not only to understand the probable link between hypothyroidism and AD but also in providing the basis for future prevention and treatment of AD in hypothyroid patients.</em>
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5

Gordon, C. Ibeanu. "Analysis of Gene Expression and Neuronal Phenotype in Neuroscreen-1 (NS-1) Cells." International Journal of Biomedical Investigation 1, no. 3 (2018): 1–13. https://doi.org/10.31531/2581-4745.1000115.

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<em>Neuroscreen-1 (NS-1) a sub-clone of pheochromocytoma (PC12) cell is gaining broad acceptance as in vitro neuronal model for biochemical and phenotypic assays due to robust growth and differentiation profiles. However, the molecular characteristics of the cell remains to be documented. In this study, we performed comparative analysis for expression of neuronal marker genes in undifferentiated and nerve growth factor (NGF) differentiated NS-1 and PC12 by qPCR and immunoblot assays. We show that differentiation of NS-1 occurred under low concentrations of NGF relative to PC12. Cell growth also occurred more rapidly in NS-1.&nbsp; Transcriptional analysis of neuronal marker genes showed comparable expression of tyrosine receptor kinases (Ntrk1, Ntrk2, NGFR/p75NTR) and muscarinic acetylcholine (Chrm1, Chrm2, Chrm3, Chrm4) receptors in unspecialized cells. Ntrk2, adenosine receptors (Adora1, Adora2A) and choline acetyltransferase (ChAT) were altered in undifferentiated NS-1. In contrast, Ntrk1, Ntrk2, Chrm2 transcripts were vastly increased in NS-1 with NGF exposure, while Ntrk3, Adora1 and Adora2A transcripts were reduced. In differentiated PC12, Chrm4 and ChAT were markedly upregulated. Our data suggests that differences in morphological and phenotypic characteristics that distinguish NS-1 from PC12 is likely the product of altered gene expression. Furthermore, expression of neuron type genes in NS-1 support its use as an alternative model to PC12.</em>
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6

Ibeanu, Gordon C. "Analysis of Gene Expression and Neuronal Phenotype in Neuroscreen-1 (NS-1) Cells." International Journal of Biomedical Investigation 1, no. 3 (2018): 1–13. http://dx.doi.org/10.31531/2581-4745.1000115.

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7

Yeyeodu, Susan, Nailya Gilyazova, Eun Young Huh, et al. "A Trifluoromethyl Analog of Verbenachalcone Promotes Neurite Outgrowth and Cell Proliferation of NeuroScreen-1 Cells." Cellular and Molecular Neurobiology 31, no. 1 (2010): 145–53. http://dx.doi.org/10.1007/s10571-010-9563-3.

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8

Gopalakrishna, Rayudu, Usha Gundimeda, Sarah Zhou, et al. "Laminin-1 induces endocytosis of 67KDa laminin receptor and protects Neuroscreen-1 cells against death induced by serum withdrawal." Biochemical and Biophysical Research Communications 495, no. 1 (2018): 230–37. http://dx.doi.org/10.1016/j.bbrc.2017.11.015.

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9

Lin, Ching-Yi, Whitney J. Huang, Kevin Li, et al. "Differential intensity-dependent effects of magnetic stimulation on the longest neurites and shorter dendrites in neuroscreen-1 cells." Journal of Neural Engineering 12, no. 2 (2015): 026013. http://dx.doi.org/10.1088/1741-2560/12/2/026013.

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10

Chaurasiya, Narayan D., Surabhi Shukla, and Babu L. Tekwani. "A Combined In Vitro Assay for Evaluation of Neurotrophic Activity and Cytotoxicity." SLAS DISCOVERY: Advancing the Science of Drug Discovery 22, no. 6 (2017): 667–75. http://dx.doi.org/10.1177/2472555217698677.

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Neurotrophic assays are phenotypic methods to identify molecules that stimulate differentiation of neuronal cells. Bioactive small molecules with neurotrophic actions hold great promise as therapeutic agents for the treatment of neurodegenerative diseases and neuronal injuries by virtue of their ability to stimulate neuritic outgrowth. A combined in vitro method, which measures neurotrophic activity and cytotoxicity in a single assay, has been described. This assay, performed in 96-well microplates with PC12 and Neuroscreen-1 (NS-1; a subclone of PC12) cells, is a simple tool for identification of new neurotrophic agents. Stimulation of neurite outgrowth was measured with NIS software by analysis of digital cell images as multiple parameters, namely, mean neurite length, neurite length/cell, nodes/cell, and number of neurites/cell. The assay has been standardized and validated with dose-response analysis for nerve growth factor (NGF) and mechanism-based inhibitors of NGF-induced neurite outgrowth, namely, SU6656 (an Src family kinase inhibitor) and PD98059 (a MEK inhibitor). The assay has been successfully applied for screening natural and synthetic compound libraries for cytotoxicity and neurotrophic activity. Screening of a set of harmala alkaloids identified harmine as a potential neurotrophic molecule that significantly stimulated NGF-induced neurite outgrowth in the NS-1 cells. Important advantages of this method are its simplicity and determination of cytotoxicity and neurotrophic activity in a single assay. This assay may be suitable for primary and cultured neuronal cells.
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11

Komatsu, Masaaki, Heather E. Wheeler, Claudia Wing, Shannon Delaney, and M. Eileen Dolan. "Identification of pharmacogenetic target genes associated with chemotherapy-induced peripheral neuropathy." Journal of Clinical Oncology 31, no. 15_suppl (2013): e13541-e13541. http://dx.doi.org/10.1200/jco.2013.31.15_suppl.e13541.

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e13541 Background: Chemotherapy-induced Peripheral Neuropathy (CIPN) is a dose-limiting toxicity of many anticancer drugs, including taxanes, vinca alkaloids and platinating agents. There are no effective means to predict which patients are at risk for CIPN nor are there effective treatments. Therefore, we have used a whole genome approach in lymphoblastoid cell lines (LCLs) alone or combined with clinical study data to identify target genes associated with CIPN that we tested further in Neuroscreen-1 (NS-1: rat pheochromocytoma) cells and human neurons derived from induced pluripotent stem cells (iCell neurons). Methods: For selecting candidate target genes, we compared published genome-wide association studies (GWAS) results of sensory peripheral neuropathy in cancer patients receiving paclitaxel with GWAS results of pharmacologic phenotypes in LCLs after paclitaxel treatment. We evaluated neurotoxic drug-induced cell growth inhibition using Cell Titer Glo and neurite parameters of outgrowth, processes and branching through imaging following transient knockdown of target genes in NS-1 cells and iCell neurons. Results: After treatment of paclitaxel, vincristine and cisplatin, iCell neurons showed decrease of neurite outgrowth, processes and branching. Whereas, these parameters did not decrease for hydroxyurea, which is not reported to induce neuropathy. Through transient knockdown in NS-1 cells of RFX2, a gene identified through clinical and preclinical studies to be associated with paclitaxel-induced neuropathy, sensitivity to paclitaxel increased as measured by a decrease of neurite outgrowth. Conclusions: These findings imply our model system could identify pharmacogenetic target genes associated with CIPN. These genes have potential for biomarkers or therapeutic targets and enable future personalized treatments for cancer patients at risk for CIPN.
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12

Mwangala, Patrick N., Charles R. Newton, Melanie Abas, and Amina Abubakar. "Screening tools for HIV-associated neurocognitive disorders among adults living with HIV in sub-Saharan Africa: A scoping review." AAS Open Research 1 (November 19, 2018): 28. http://dx.doi.org/10.12688/aasopenres.12921.1.

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Background: People living with HIV are at risk of developing HIV-associated neurocognitive disorders (HAND) which adversely affects their quality of life. Routine screening of HAND in HIV care is recommended to identify subtle changes in cognitive functioning and allow for early interventions. However, HAND detection is rarely done in sub-Saharan Africa (SSA), partly due to a lack of adequately standardized screening tools. This review was conducted to identify the commonly used screening tools for HAND in SSA and document their psychometric properties and diagnostic accuracy.Methods:We searched Ovid Medline, PsycINFO and Web of Sciences databases for empirical studies published from 1/1/1980 to 31/8/2018 on HAND among adults living with HIV in SSA.Results:We identified 14 eligible studies, of which 9 were from South Africa. The International HIV Dementia Scale (IHDS) was the most frequently reported tool, being used in more than half of the studies. However most studies only reported the diagnostic accuracy of this and other tools, with specificity ranging from 37% to 81% and sensitivity ranging from 45% to 100%. Appropriate data on construct validity and reliability of tools was rarely documented. Although most tools performed well in screening for severe forms of HAND, they lacked sensitivity and specificity for moderate forms of HAND. NeuroScreen, one of the newer tools, yielded good diagnostic accuracy in its initial evaluation in South Africa (81% to 93% sensitivity and 71% to 81% specificity).Conclusions:This review identified a lack of adequately standardized and contextually relevant HAND screening tools in SSA. Most screening tools for HAND used in SSA possess inadequate psychometric properties and diagnostic accuracy. There is a need for further validation of existing tools and development of new tools to make them sensitive and specific enough to identify both severe and moderate forms of HAND in SSA.
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13

Mwangala, Patrick N., Charles R. Newton, Melanie Abas, and Amina Abubakar. "Screening tools for HIV-associated neurocognitive disorders among adults living with HIV in sub-Saharan Africa: A scoping review." AAS Open Research 1 (October 30, 2019): 28. http://dx.doi.org/10.12688/aasopenres.12921.2.

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Background: People living with HIV are at risk of developing HIV-associated neurocognitive disorders (HAND) which adversely affects their quality of life. Routine screening of HAND in HIV care is recommended to identify clinically important changes in cognitive functioning and allow for early interventions. However, HAND detection in routine clinical practice has never been reported in sub-Saharan Africa (SSA), partly due to a lack of adequately standardized screening tools. This review was conducted to identify the commonly used screening tools for HAND in SSA and document their psychometric properties and diagnostic accuracy. Methods: We searched Ovid Medline, PsycINFO and Web of Sciences databases for empirical studies published from 1/1/1980 to 31/8/2018 on HAND among adults living with HIV in SSA. Results: We identified 14 eligible studies, of which 9 were from South Africa. The International HIV Dementia Scale (IHDS) was the most frequently reported tool, being used in more than half of the studies. However most studies only reported the diagnostic accuracy of this and other tools, with specificity ranging from 37% to 81% and sensitivity ranging from 45% to 100%. Appropriate data on construct validity and reliability of tools was rarely documented. Although most tools performed well in screening for severe forms of HAND, they lacked sensitivity and specificity for mild forms of HAND. NeuroScreen, one of the newer tools, yielded good diagnostic accuracy in its initial evaluation in South Africa (81% to 93% sensitivity and 71% to 81% specificity). Conclusions: This review identified a lack of adequately standardized and contextually relevant HAND screening tools in SSA. Most screening tools for HAND used in SSA possess inadequate psychometric properties and diagnostic accuracy. There is a need for further validation of existing tools and development of new HAND screening tools in SSA.
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14

Chua, PinFen, and William K. Lim. "Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents." Scientific Reports 11, no. 1 (2021). http://dx.doi.org/10.1038/s41598-021-87431-4.

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AbstractStroke causes death and disability globally but no neuroprotectant is approved for post-stroke neuronal injury. Neuroprotective compounds can be identified using oxygen glucose deprivation (OGD) of neuronal cells as an in vitro stroke model. Nerve growth factor (NGF)-differentiated PC12 pheochromocytoma cells are frequently used. However, investigators often find their clonal variant undifferentiable and are uncertain of optimal culture conditions. Hence we studied 3 commonly used PC12 variants: PC12 Adh, PC12 from Riken Cell Bank (PC12 Riken) and Neuroscreen-1 (NS-1) cells. We found DMEM the optimal media for PC12 Riken and NS-1 cells. Using a novel serum-free media approach, we identified collagen IV as the preferred adhesive substrate for both cell lines. We found PC12 Adh cells cannot attach without serum and is unable to differentiate using NGF. NS-1 cells differentiated to a maximal 72.7 ± 5.2% %, with substantial basal differentiation. We optimised differentiated NS-1 cells for an in vitro stroke model using 3 h of OGD resulting in ~ 70% viable cells. We screened 5 reported neuroprotectants and provide the first report that serotonin is antiapoptotic in a stroke model and the 5-HT1Aagonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) is neuroprotective in PC12 cells. Thus we demonstrate the optimisation and validation for a PC12 cell-based in vitro stroke model.
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Gopalakrishna, Rayudu, Angela Zhu, Andrew Oh, et al. "Abstract WP141: Cyclic-AMP Induces Nogo-A Receptor NgR1 Internalization and Inhibits Nogo-A-Mediated Collapse of Growth Cone." Stroke 51, Suppl_1 (2020). http://dx.doi.org/10.1161/str.51.suppl_1.wp141.

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Recovery of stroke and neuronal injuries requires the promotion of axonal regeneration from the remaining neurons. However, axonal regeneration is inhibited by diverse axonal growth inhibitors, such as Nogo-A. C-terminal domain of Nogo-A, Nogo-66 binds to the Nogo-A receptor 1 (NgR1) and induces the collapse of growth cones and inhibition of neurite outgrowth. NgR1 is also a receptor for additional axonal growth inhibitors. In this study, by using indirect immunofluorescence and biotinylation method, we have found that a cell-permeable cAMP analog (dibutyryl-cAMP) and other intracellular cAMP-elevating agents, such as forskolin, which directly activates adenylyl cyclase, and rolipram, which inhibits cyclic nucleotide phosphodiesterase, all induced rapid internalization of the cell surface NgR1 in Neuroscreen-1 (NS-1) cells. This endocytosis of NgR1 is lipid raft mediated. These cAMP-elevating agents induced a reversible distribution of NgR1 between the cell surface and intracellular compartment; NgR1 distributed to the cell surface at low levels of cAMP and distributed to an intracellular compartment at high levels of cAMP. Using pharmacological activators and inhibitors of protein kinase A (PKA) and the exchange protein directly activated by cAMP (Epac), we found that NgR1 internalization is independent of PKA but dependent on Epac. There is a correlation between the decrease in cell surface expression of NgR1 decreased sensitivity of NS-1 cells to Nogo-66-induced growth cone collapse. Therefore, besides axonal growth inhibitors affecting neurons, neurons by themselves self-regulate their own sensitivity to extracellular cues such as axonal growth inhibitors. This normal cellular regulatory mechanism may be therapeutically applied to overcome axonal growth inhibitors and enhance functional recovery after stroke and neuronal injuries.
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16

Tekwani, BL, ND Chaurasiya, S. Shukla, I. Muhammad, and LA Walker. "Neurotrophic and neuritogenic drug leads from natural products: A combined in vitro assay for evaluation of cell viability and NGF-stimulated neuritic outgrowth in neuroscreen 1 cells." Planta Medica 81, no. 11 (2015). http://dx.doi.org/10.1055/s-0035-1556135.

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17

GOPALAKRISHNA, Rayudu, Charlotte Y. Lin, Andrew Oh, and William J. Mack. "Abstract P803: Pituitary Adenylate Cyclase-Activating Polypeptide via Cyclic-AMP Inhibits Nogo-A by Internalizing Its Receptor NgR1." Stroke 52, Suppl_1 (2021). http://dx.doi.org/10.1161/str.52.suppl_1.p803.

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Introduction: After a stroke, axonal regeneration is inhibited by diverse axonal growth inhibitors, such as Nogo-A. They bind to the Nogo-A receptor 1 (NgR1) and induce the collapse of growth cones and inhibit neurite outgrowth. Since NgR1 is the receptor for a variety of axonal growth inhibitors, it is a crucial target for the prevention of axonal growth inhibition. Pituitary adenylate cyclase-activating polypeptide (PACAP) has neuroprotective and neurotrophic activities and increases neuritogenesis and synaptic plasticity. It enhances functional recovery after stroke in various animal models. Methods: Neuroscreen-1 (NS-1) cells were selected for this study as they produce rapid and robust neurite outgrowth with NGF. Cell surface NgR1 was detected using the indirect immunofluorescence method. The internalization of NgR1 was quantitated using the biotinylation method and Western immunoblotting. Results: Using the indirect immunofluorescence method, we found that PACAP (PACAP-38) induced a rapid decrease in the cell surface expression of NgR1 in NS-1 cells. The biotinylation method revealed that PACAP induced the internalization of NgR1. This internalization of NgR1 was blocked by pretreatment of NS-1 cells with SQ 22536, an inhibitor for adenylate cyclase, suggesting that cAMP plays a crucial role in the internalization of NgR1. The protein kinase A (PKA)-specific inhibitor KT5720 did not block PACAP-induced NgR1 internalization, whereas the exchange protein directly activated by cAMP (Epac)-specific inhibitor ESI-09 blocked this internalization. Collectively, this data suggests that PACAP-induced NgR1 internalization is independent of PKA but is dependent on Epac. The PACAP-induced decrease in cell surface expression of NgR1 and its internalization desensitized NS-1 cells to Nogo-66-induced growth cone collapse and enhanced neuritogenesis. Conclusion: Cyclic-AMP and Epac are involved in the PACAP-induced desensitization of neuronal cells to Nogo-A and increase in neuritogenesis. Since PACAP crosses the blood-brain barrier, it may be a useful therapeutic agent to overcome axonal growth inhibitors and enhance functional recovery after stroke.
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