Yaz Kisanuki - Academia.edu (original) (raw)

Papers by Yaz Kisanuki

Japanese Society of Neurology, May 8, 2019

Introduction Mesenchymal stem cells (MSCs) are defined as a population of multipotent cells able ... more Introduction Mesenchymal stem cells (MSCs) are defined as a population of multipotent cells able to differentiate and produce any cell type needed in a repair process, such as osteoblasts, chondroblasts, neurons, epithelial cells, and cardiac cells (1,2). This cell type has become the focus of numerous studies worldwide for providing clinically promising perspectives for cell therapy and also for its immunomodulatory potential (3,4), although the mechanisms of immunosuppression on inflammatory response and the mechanisms of transplant rejection are not fully elucidated (5). Recent studies have described the use of allogeneic and autologous MSCs for the repair of various tissues (4,6). However, there is little research involving xenotransplantation in animals and most of them only evaluated the cellular interaction in vitro between MSCs and T lymphocytes. Because of the great therapeutic potential of MSCs, in addition to the persistent doubts about their immunosuppressive capacity in vivo, further studies are needed to investigate the real potential of xenogenic transplantation using these cells for tissue repair in animals. Therefore, this study evaluated clinical and radiographic aspects of xenogenic transplantation of rat bone marrow-derived MSCs for the repair of radial bone defects created in rabbits. 2. Materials and methods This study was approved by the Institutional Animal Care Committee (protocol 97/2010). 2.1 Cellular culture A total of five male, 4-week old Wistar rats were euthanized using anesthetic overdose. The animals were immersed in alcohol 70° to ensure antisepsis for cell collection and were taken to the laminar flow cabinet. The femurs were disarticulated and removed aseptically. The distal epiphyses were cut and the medullary canal was flushed with Dulbecco's modified Eagle's medium (DMEM, Gibco, Grand Island, NY, USA) with low glucose, containing 10% fetal bovine serum (FBS) (Gibco), 50.0 mg L-1 gentamicin,

Scientific Reports, Nov 3, 2017

Endothelin 1 (ET-1), mainly produced from vascular endothelial cells, induces vasoconstriction in... more Endothelin 1 (ET-1), mainly produced from vascular endothelial cells, induces vasoconstriction in physiological conditions. The endothelin receptor antagonist is among the most effective agents for pulmonary hypertension. However, little is known about the production source of ET-1 in inflammation and immunity. Here, we studied whether T cell-mediated ET-1 production system exists and operates independent of the production system in vascular endothelial cells. ET-1 production was readily detectable in the culture supernatant of human PBMCs and murine spleen cells stimulated with anti-CD3 antibody. Immunocytostaining showed that ET-1-producing cells emerged only in PBMCs stimulated with anti-CD3 antibody. Using the Transwell system, both murine and human monocytes sorted with magnetic beads in the inner chamber produced ET-1 when T cells were activated with antigen or anti-CD3 antibody in the outer chamber. This ET-1 production was inhibited by anti-IFN-γ and/or TNF-α antibody. Furthermore, monocytes purified from ET flox/flox ;Tie2-Cre(+) mice, which conditionally lack ET-1 in hematopoietic stem cells and vascular endothelial cells, did not produce ET-1 even when stimulated by antigen-specific T cell activation. This study demonstrates the existence of an immune-mediated ET-1 production induced by T cells upon activation through IFN-γ and TNF-α.

Molecular Genetics and Metabolism, Feb 1, 2022

Journal of Clinical Investigation, May 1, 2003

The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: in... more The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: insulin receptor (IR); endothelin-1 (ET-1); vascular endothelial cell insulin receptor knockout (VENIRKO); glucose-tolerance test (GTT); insulintolerance tests (ITT); systolic blood pressure (SBP); mean blood pressure (MBP); diastolic blood pressure (DBP); heart rate (HR); glucose infusion rate (GIR); lipoprotein lipase (LPL).

Japanese Circulation Journal-english Edition, 2007

The Journal of Neuroscience, Apr 27, 2011

Orexin-A and orexin-B are hypothalamic neuropeptides that play critical roles in the maintenance ... more Orexin-A and orexin-B are hypothalamic neuropeptides that play critical roles in the maintenance of wakefulness. Intracerebroventricular (ICV) administration of orexin-A has been shown to promote wakefulness and suppress both rapid eye movement (REM) sleep and non-REM (NREM) sleep through the orexin receptor-1 (OX 1 R) and orexin receptor-2 (OX 2 R). Here, we elucidated the differential roles of orexin receptors in the regulation of sleep and wakefulness by comparing the effects of ICV orexin-A administration in wild-type, OX 1 R Ϫ/Ϫ , and OX 2 R Ϫ/Ϫ mice. The effects of orexin-A on wakefulness and NREM sleep were significantly attenuated in both knockout mice as compared with wild-type mice, with substantially larger attenuation in OX 2 R Ϫ/Ϫ mice than in OX 1 R Ϫ/Ϫ mice. These results suggest that although the OX 2 R-mediated pathway has a pivotal role in the promotion of wakefulness, OX 1 R also plays additional roles in promoting arousal. In contrast, suppression of REM sleep by orexin-A administration was slightly and similarly attenuated in both OX 1 R Ϫ/Ϫ and OX 2 R Ϫ/Ϫ mice, suggesting a comparable contribution of the two receptors to REM sleep suppression. Histological studies demonstrated differential distributions of each receptor subtype in distinct neuronal populations with specific neurotransmitter identities in brainstem cholinergic/monoaminergic neurons. In the laterodorsal tegmental and pedunculopontine tegmental nuclei especially, cholinergic neurons exclusively expressed OX 1 R mRNA, but OX 2 R mRNA was expressed mainly in GABAergic putative interneurons. Thus, each orexin receptor subtype plays differential roles in gating NREM and REM sleep through distinct neuronal pathways.

Japanese Circulation Journal-english Edition, Mar 1, 2004

Infection and Immunity, Apr 1, 2005

Journal of Clinical Investigation, Aug 15, 2001

The thrombomodulin (TM) gene was ablated in mice in a cell type-restricted manner from vascular e... more The thrombomodulin (TM) gene was ablated in mice in a cell type-restricted manner from vascular endothelium by Cre-recombinase-mediated excision controlled by the endothelial cell lineage-specific Tie2 promoter. Forty percent of mutant (TMLox-) mice display a distinct lethal embryonic phenotype not observed in completely TM-deficient embryos. The remaining 60% of TMLox mice survive beyond birth, but invariably succumb to a severe hypercoagulable state and massive thrombosis after 3 weeks, terminating in a lethal consumptive coagulopathy. The progression of thrombosis was age-and sex-dependent. Disruption of the TM/protein C pathway was not associated with a latent proinflammatory state. Disease onset and progression could be prevented by warfarin anticoagulation. These results show that in mice, loss of endothelial cell TM function causes spontaneous and fatal thrombosis in the arterial and venous circulation, resulting from unfettered activation of the coagulation system. The combination of complete disease penetrance, uniform disease onset at young age, large vessel thrombosis of the extremities and multiple organ systems, and consumptive coagulopathy as the disease end-point provides a unique mouse model of human thrombotic disease.

Neurology, Feb 12, 2018

Go to Neurology.org/N for full disclosures. Funding information and disclosures deemed relevant b... more Go to Neurology.org/N for full disclosures. Funding information and disclosures deemed relevant by the authors, if any, are provided at the end of the article.

Rivista Di Neuroradiologia, Jan 7, 2019

Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a rare white m... more Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a rare white matter degenerative disease characterized by both axonal and glial injury due to a defect in the CSF1R gene. In this report, we describe ALSP in a previously healthy 40-year-old woman presenting with insidiously progressive confusion, memory loss, and loss of social inhibitions. Characteristic magnetic resonance imaging findings for ALSP elucidated the diagnosis, including chronic foci of diffusion restriction in a non-vascular distribution, lack of temporal/infratentorial involvement, cortical sparing, and lack of enhancement. CSF1R genetic testing further confirmed the diagnosis and the patient underwent supportive medical management for symptom control. ALSP can pose a unique diagnostic challenge given its particular adult-onset presentation, but early recognition is key given the poor prognosis and the potential for family genetic testing.

Frontiers in Neuroscience, 2013

Orexin neuropeptides influence multiple homeostatic functions and play an essential role in the e... more Orexin neuropeptides influence multiple homeostatic functions and play an essential role in the expression of normal sleep-wake behavior. While their two known receptors (OX 1 and OX 2) are targets for novel pharmacotherapeutics, the actions mediated by each receptor remain largely unexplored. Using brain slices from mice constitutively lacking either receptor, we used whole-cell and Ca 2+ imaging methods to delineate the cellular actions of each receptor within cholinergic [laterodorsal tegmental nucleus (LDT)] and monoaminergic [dorsal raphe (DR) and locus coeruleus (LC)] brainstem nuclei-where orexins promote arousal and suppress REM sleep. In slices from OX −/ 2 − mice, orexin-A (300 nM) elicited wild-type responses in LDT, DR, and LC neurons consisting of a depolarizing current and augmented voltage-dependent Ca 2+ transients. In slices from OX −/ 1 − mice, the depolarizing current was absent in LDT and LC neurons and was attenuated in DR neurons, although Ca 2+-transients were still augmented. Since orexin-A produced neither of these actions in slices lacking both receptors, our findings suggest that orexin-mediated depolarization is mediated by both receptors in DR, but is exclusively mediated by OX 1 in LDT and LC neurons, even though OX 2 is present and OX 2 mRNA appears elevated in brainstems from OX −/ 1 − mice. Considering published behavioral data, these findings support a model in which orexin-mediated excitation of mesopontine cholinergic and monoaminergic neurons contributes little to stabilizing spontaneous waking and sleep bouts, but functions in context-dependent arousal and helps restrict muscle atonia to REM sleep. The augmented Ca 2+ transients produced by both receptors appeared mediated by influx via L-type Ca 2+ channels, which is often linked to transcriptional signaling. This could provide an adaptive signal to compensate for receptor loss or prolonged antagonism and may contribute to the reduced severity of narcolepsy in single receptor knockout mice.

Japanese Circulation Journal-english Edition, Mar 1, 2005

Proceedings of the National Academy of Sciences of the United States of America, Feb 5, 2004

Journal of Clinical Investigation, May 1, 2003

The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: in... more The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: insulin receptor (IR); endothelin-1 (ET-1); vascular endothelial cell insulin receptor knockout (VENIRKO); glucose-tolerance test (GTT); insulintolerance tests (ITT); systolic blood pressure (SBP); mean blood pressure (MBP); diastolic blood pressure (DBP); heart rate (HR); glucose infusion rate (GIR); lipoprotein lipase (LPL).

The Cerebellum, Apr 5, 2023

To accelerate and facilitate clinical trials, the Ataxia Global Initiative (AGI) was established ... more To accelerate and facilitate clinical trials, the Ataxia Global Initiative (AGI) was established as a worldwide research platform for trial readiness in ataxias. One of AGI's major goals is the harmonization and standardization of outcome assessments. Clinical outcome assessments (COAs) that describe or reflect how a patient feels or functions are indispensable for clinical trials, but similarly important for observational studies and in routine patient care. The AGI working group on COAs has defined a set of data including a graded catalog of COAs that are recommended as a standard for future assessment and sharing of clinical data and joint clinical studies. Two datasets were defined: a mandatory dataset (minimal dataset) that can ideally be obtained during a routine clinical consultation and a more demanding extended dataset that is useful for research purposes. In the future, the currently most widely used clinician-reported outcome measure (ClinRO) in ataxia, the scale for the assessment and rating of ataxia (SARA), should be developed into a generally accepted instrument that can be used in upcoming clinical trials. Furthermore, there is an urgent need (i) to obtain more data on ataxia-specific, patient-reported outcome measures (PROs), (ii) to demonstrate and optimize sensitivity to change of many COAs, and (iii) to establish methods and evidence of anchoring change in COAs in patient meaningfulness, e.g., by determining patient-derived minimally meaningful thresholds of change. Keywords Activities of daily living (ADL) • Ataxia • Clinical outcome assessment (COA) • Scale for the assessment and rating of ataxia (SARA) • Standardization Ataxias, which have long been considered untreatable, are now becoming models for the development of targeted therapies. Consequently, an increasing number of observational studies and clinical trials is expected within the next years [1]. This situation requires harmonization and consensus on the use of appropriate clinical outcome assessments (COAs). In this manuscript, we provide (1) an overview of available COAs and (2) a proposal for the data content of ataxia registries based on the consensus of the Ataxia Global Initiative (AGI) working group on COAs.

Journal of the American College of Cardiology, Mar 1, 2002

As compared to patients of Group 1, those of Group 2 were younger (47±14 vs 41±15 years, p<0.001)... more As compared to patients of Group 1, those of Group 2 were younger (47±14 vs 41±15 years, p<0.001), and showed less advanced LV dysfunction (EF 28±9 vs 35±9%, p<0.001) and remodelling (end-diastolic diameter index 38±7 vs 35±7 minim2, p<0.001) at the time of diagnosis. Most patients of Group 1 and 2 were treated with ACE-I (93% and 88%, p=NS) and BB (83% and 74%, p<0.05). Five and 10-year transplant-free survival was respectively 73 and 57% in Group1 vs 93 and 86% in Group 2, while hospitatisation-free survival was 47 and 32% in Group1 vs 70 and 57% in Group 2 (both p<0.001). No outcome difference was observed between Group 2a and 2b, whereas 5 and 10-year transplant-free survival was significantly better in patients of Group 2 who were treated with BB than in those not receiving BB (p=0.007 after stratification for the severity of the disease). At 6 to 8 years of follow-up, 38% of patients of Group 2 developed HF symptoms, and/or a decrease of LVEF>10%, and/or the need of hospitalisation for cardiovascular reasons. The long-term progression of HF symptoms and LV dysfunction was similar in asymptomatic patients of Group 2a and 2b and not significantly different to that of patients of Group 1. Even though asymptomatic DC patients receiving an optimal medical treatment are characterized by low rates of death or heart transplant, nevertheless during a long-term follow-up they frequently exhibit a worsening of clinical status and LV function. Our data suggest that in these patients an early and aggressive BB strategy should be carefully considered in order to counteract as much as possible the Ioeg-term progression of the disease.

European Journal of Neuroscience, Jun 22, 2010

Pharmacological studies of narcoleptic canines indicate that exaggerated pontine cholinergic tran... more Pharmacological studies of narcoleptic canines indicate that exaggerated pontine cholinergic transmission promotes cataplexy. Since disruption of orexin (hypocretin) signaling is a primary defect in narcolepsy with cataplexy, we investigated whether markers of cholinergic synaptic transmission might be altered in mice constitutively lacking orexin receptors (double receptor knockout; DKO). We found that choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT) and the high-affinity choline transporter (CHT1) but not acetylcholinesterase (AChE) were significantly higher in samples from DKO compared to wild-type (WT) mice. This was region-specific since levels were elevated in samples from the laterodorsal tegmental nucleus (LDT) and the fifth motor nucleus (Mo5) but not in whole brainstem samples. Consistent with region-specific changes, we were unable to detect significant differences in Western blots for ChAT and CHT1 in isolates from brainstem, thalamus and cortex or in ChAT enzymatic activity in the pons. However, using ChAT immunocytochemistry, we found that while the number of cholinergic neurons in the LDT and Mo5 were not different, the intensity of somatic ChAT immunostaining was significantly greater in the LDT, but not Mo5, from DKOs compared to WTs. We also found that ChAT activity was significantly reduced in cortical samples from DKOs compared to WTs. Collectively, these findings suggest that the orexins can regulate neurotransmitter expression and that the constitutive absence of orexin signaling results in an upregulation of the machinery necessary for cholinergic neurotransmission in a mesopontine population of neurons that have been associated with both normal REM sleep and cataplexy.

PLOS ONE, Apr 13, 2011

To investigate how cholinergic systems regulate aspects of the sleep disorder narcolepsy, we vide... more To investigate how cholinergic systems regulate aspects of the sleep disorder narcolepsy, we video-monitored mice lacking both orexin (hypocretin) receptors (double knockout; DKO mice) while pharmacologically altering cholinergic transmission. Spontaneous behavioral arrests in DKO mice were highly similar to those reported in orexin-deficient mice and were never observed in wild-type (WT) mice. A survival analysis revealed that arrest lifetimes were exponentially distributed indicating that random, Markovian processes determine arrest lifetime. Low doses (0.01, 0.03 mg/kg, IP), but not a high dose (0.08 mg/ kg, IP) of the cholinesterase inhibitor physostigmine increased the number of arrests but did not alter arrest lifetimes. The muscarinic antagonist atropine (0.5 mg/kg, IP) decreased the number of arrests, also without altering arrest lifetimes. To determine if muscarinic transmission in pontine areas linked to REM sleep control also influences behavioral arrests, we microinjected neostigmine (50 nl, 62.5 mM) or neostigmine + atropine (62.5 mM and 111 mM respectively) into the nucleus pontis oralis and caudalis. Neostigmine increased the number of arrests in DKO mice without altering arrest lifetimes but did not provoke arrests in WT mice. Co-injection of atropine abolished this effect. Collectively, our findings establish that behavioral arrests in DKO mice are similar to those in orexin deficient mice and that arrests have exponentially distributed lifetimes. We also show, for the first time in a rodent narcolepsy model, that cholinergic systems can regulate arrest dynamics. Since perturbations of muscarinic transmission altered arrest frequency but not lifetime, our findings suggest cholinergic systems influence arrest initiation without influencing circuits that determine arrest duration.

Japanese Society of Neurology, May 8, 2019

Introduction Mesenchymal stem cells (MSCs) are defined as a population of multipotent cells able ... more Introduction Mesenchymal stem cells (MSCs) are defined as a population of multipotent cells able to differentiate and produce any cell type needed in a repair process, such as osteoblasts, chondroblasts, neurons, epithelial cells, and cardiac cells (1,2). This cell type has become the focus of numerous studies worldwide for providing clinically promising perspectives for cell therapy and also for its immunomodulatory potential (3,4), although the mechanisms of immunosuppression on inflammatory response and the mechanisms of transplant rejection are not fully elucidated (5). Recent studies have described the use of allogeneic and autologous MSCs for the repair of various tissues (4,6). However, there is little research involving xenotransplantation in animals and most of them only evaluated the cellular interaction in vitro between MSCs and T lymphocytes. Because of the great therapeutic potential of MSCs, in addition to the persistent doubts about their immunosuppressive capacity in vivo, further studies are needed to investigate the real potential of xenogenic transplantation using these cells for tissue repair in animals. Therefore, this study evaluated clinical and radiographic aspects of xenogenic transplantation of rat bone marrow-derived MSCs for the repair of radial bone defects created in rabbits. 2. Materials and methods This study was approved by the Institutional Animal Care Committee (protocol 97/2010). 2.1 Cellular culture A total of five male, 4-week old Wistar rats were euthanized using anesthetic overdose. The animals were immersed in alcohol 70° to ensure antisepsis for cell collection and were taken to the laminar flow cabinet. The femurs were disarticulated and removed aseptically. The distal epiphyses were cut and the medullary canal was flushed with Dulbecco's modified Eagle's medium (DMEM, Gibco, Grand Island, NY, USA) with low glucose, containing 10% fetal bovine serum (FBS) (Gibco), 50.0 mg L-1 gentamicin,

Scientific Reports, Nov 3, 2017

Endothelin 1 (ET-1), mainly produced from vascular endothelial cells, induces vasoconstriction in... more Endothelin 1 (ET-1), mainly produced from vascular endothelial cells, induces vasoconstriction in physiological conditions. The endothelin receptor antagonist is among the most effective agents for pulmonary hypertension. However, little is known about the production source of ET-1 in inflammation and immunity. Here, we studied whether T cell-mediated ET-1 production system exists and operates independent of the production system in vascular endothelial cells. ET-1 production was readily detectable in the culture supernatant of human PBMCs and murine spleen cells stimulated with anti-CD3 antibody. Immunocytostaining showed that ET-1-producing cells emerged only in PBMCs stimulated with anti-CD3 antibody. Using the Transwell system, both murine and human monocytes sorted with magnetic beads in the inner chamber produced ET-1 when T cells were activated with antigen or anti-CD3 antibody in the outer chamber. This ET-1 production was inhibited by anti-IFN-γ and/or TNF-α antibody. Furthermore, monocytes purified from ET flox/flox ;Tie2-Cre(+) mice, which conditionally lack ET-1 in hematopoietic stem cells and vascular endothelial cells, did not produce ET-1 even when stimulated by antigen-specific T cell activation. This study demonstrates the existence of an immune-mediated ET-1 production induced by T cells upon activation through IFN-γ and TNF-α.

Molecular Genetics and Metabolism, Feb 1, 2022

Journal of Clinical Investigation, May 1, 2003

The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: in... more The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: insulin receptor (IR); endothelin-1 (ET-1); vascular endothelial cell insulin receptor knockout (VENIRKO); glucose-tolerance test (GTT); insulintolerance tests (ITT); systolic blood pressure (SBP); mean blood pressure (MBP); diastolic blood pressure (DBP); heart rate (HR); glucose infusion rate (GIR); lipoprotein lipase (LPL).

Japanese Circulation Journal-english Edition, 2007

The Journal of Neuroscience, Apr 27, 2011

Orexin-A and orexin-B are hypothalamic neuropeptides that play critical roles in the maintenance ... more Orexin-A and orexin-B are hypothalamic neuropeptides that play critical roles in the maintenance of wakefulness. Intracerebroventricular (ICV) administration of orexin-A has been shown to promote wakefulness and suppress both rapid eye movement (REM) sleep and non-REM (NREM) sleep through the orexin receptor-1 (OX 1 R) and orexin receptor-2 (OX 2 R). Here, we elucidated the differential roles of orexin receptors in the regulation of sleep and wakefulness by comparing the effects of ICV orexin-A administration in wild-type, OX 1 R Ϫ/Ϫ , and OX 2 R Ϫ/Ϫ mice. The effects of orexin-A on wakefulness and NREM sleep were significantly attenuated in both knockout mice as compared with wild-type mice, with substantially larger attenuation in OX 2 R Ϫ/Ϫ mice than in OX 1 R Ϫ/Ϫ mice. These results suggest that although the OX 2 R-mediated pathway has a pivotal role in the promotion of wakefulness, OX 1 R also plays additional roles in promoting arousal. In contrast, suppression of REM sleep by orexin-A administration was slightly and similarly attenuated in both OX 1 R Ϫ/Ϫ and OX 2 R Ϫ/Ϫ mice, suggesting a comparable contribution of the two receptors to REM sleep suppression. Histological studies demonstrated differential distributions of each receptor subtype in distinct neuronal populations with specific neurotransmitter identities in brainstem cholinergic/monoaminergic neurons. In the laterodorsal tegmental and pedunculopontine tegmental nuclei especially, cholinergic neurons exclusively expressed OX 1 R mRNA, but OX 2 R mRNA was expressed mainly in GABAergic putative interneurons. Thus, each orexin receptor subtype plays differential roles in gating NREM and REM sleep through distinct neuronal pathways.

Japanese Circulation Journal-english Edition, Mar 1, 2004

Infection and Immunity, Apr 1, 2005

Journal of Clinical Investigation, Aug 15, 2001

The thrombomodulin (TM) gene was ablated in mice in a cell type-restricted manner from vascular e... more The thrombomodulin (TM) gene was ablated in mice in a cell type-restricted manner from vascular endothelium by Cre-recombinase-mediated excision controlled by the endothelial cell lineage-specific Tie2 promoter. Forty percent of mutant (TMLox-) mice display a distinct lethal embryonic phenotype not observed in completely TM-deficient embryos. The remaining 60% of TMLox mice survive beyond birth, but invariably succumb to a severe hypercoagulable state and massive thrombosis after 3 weeks, terminating in a lethal consumptive coagulopathy. The progression of thrombosis was age-and sex-dependent. Disruption of the TM/protein C pathway was not associated with a latent proinflammatory state. Disease onset and progression could be prevented by warfarin anticoagulation. These results show that in mice, loss of endothelial cell TM function causes spontaneous and fatal thrombosis in the arterial and venous circulation, resulting from unfettered activation of the coagulation system. The combination of complete disease penetrance, uniform disease onset at young age, large vessel thrombosis of the extremities and multiple organ systems, and consumptive coagulopathy as the disease end-point provides a unique mouse model of human thrombotic disease.

Neurology, Feb 12, 2018

Go to Neurology.org/N for full disclosures. Funding information and disclosures deemed relevant b... more Go to Neurology.org/N for full disclosures. Funding information and disclosures deemed relevant by the authors, if any, are provided at the end of the article.

Rivista Di Neuroradiologia, Jan 7, 2019

Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a rare white m... more Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a rare white matter degenerative disease characterized by both axonal and glial injury due to a defect in the CSF1R gene. In this report, we describe ALSP in a previously healthy 40-year-old woman presenting with insidiously progressive confusion, memory loss, and loss of social inhibitions. Characteristic magnetic resonance imaging findings for ALSP elucidated the diagnosis, including chronic foci of diffusion restriction in a non-vascular distribution, lack of temporal/infratentorial involvement, cortical sparing, and lack of enhancement. CSF1R genetic testing further confirmed the diagnosis and the patient underwent supportive medical management for symptom control. ALSP can pose a unique diagnostic challenge given its particular adult-onset presentation, but early recognition is key given the poor prognosis and the potential for family genetic testing.

Frontiers in Neuroscience, 2013

Orexin neuropeptides influence multiple homeostatic functions and play an essential role in the e... more Orexin neuropeptides influence multiple homeostatic functions and play an essential role in the expression of normal sleep-wake behavior. While their two known receptors (OX 1 and OX 2) are targets for novel pharmacotherapeutics, the actions mediated by each receptor remain largely unexplored. Using brain slices from mice constitutively lacking either receptor, we used whole-cell and Ca 2+ imaging methods to delineate the cellular actions of each receptor within cholinergic [laterodorsal tegmental nucleus (LDT)] and monoaminergic [dorsal raphe (DR) and locus coeruleus (LC)] brainstem nuclei-where orexins promote arousal and suppress REM sleep. In slices from OX −/ 2 − mice, orexin-A (300 nM) elicited wild-type responses in LDT, DR, and LC neurons consisting of a depolarizing current and augmented voltage-dependent Ca 2+ transients. In slices from OX −/ 1 − mice, the depolarizing current was absent in LDT and LC neurons and was attenuated in DR neurons, although Ca 2+-transients were still augmented. Since orexin-A produced neither of these actions in slices lacking both receptors, our findings suggest that orexin-mediated depolarization is mediated by both receptors in DR, but is exclusively mediated by OX 1 in LDT and LC neurons, even though OX 2 is present and OX 2 mRNA appears elevated in brainstems from OX −/ 1 − mice. Considering published behavioral data, these findings support a model in which orexin-mediated excitation of mesopontine cholinergic and monoaminergic neurons contributes little to stabilizing spontaneous waking and sleep bouts, but functions in context-dependent arousal and helps restrict muscle atonia to REM sleep. The augmented Ca 2+ transients produced by both receptors appeared mediated by influx via L-type Ca 2+ channels, which is often linked to transcriptional signaling. This could provide an adaptive signal to compensate for receptor loss or prolonged antagonism and may contribute to the reduced severity of narcolepsy in single receptor knockout mice.

Japanese Circulation Journal-english Edition, Mar 1, 2005

Proceedings of the National Academy of Sciences of the United States of America, Feb 5, 2004

Journal of Clinical Investigation, May 1, 2003

The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: in... more The authors have declared that no conflict of interest exists. Nonstandard abbreviations used: insulin receptor (IR); endothelin-1 (ET-1); vascular endothelial cell insulin receptor knockout (VENIRKO); glucose-tolerance test (GTT); insulintolerance tests (ITT); systolic blood pressure (SBP); mean blood pressure (MBP); diastolic blood pressure (DBP); heart rate (HR); glucose infusion rate (GIR); lipoprotein lipase (LPL).

The Cerebellum, Apr 5, 2023

To accelerate and facilitate clinical trials, the Ataxia Global Initiative (AGI) was established ... more To accelerate and facilitate clinical trials, the Ataxia Global Initiative (AGI) was established as a worldwide research platform for trial readiness in ataxias. One of AGI's major goals is the harmonization and standardization of outcome assessments. Clinical outcome assessments (COAs) that describe or reflect how a patient feels or functions are indispensable for clinical trials, but similarly important for observational studies and in routine patient care. The AGI working group on COAs has defined a set of data including a graded catalog of COAs that are recommended as a standard for future assessment and sharing of clinical data and joint clinical studies. Two datasets were defined: a mandatory dataset (minimal dataset) that can ideally be obtained during a routine clinical consultation and a more demanding extended dataset that is useful for research purposes. In the future, the currently most widely used clinician-reported outcome measure (ClinRO) in ataxia, the scale for the assessment and rating of ataxia (SARA), should be developed into a generally accepted instrument that can be used in upcoming clinical trials. Furthermore, there is an urgent need (i) to obtain more data on ataxia-specific, patient-reported outcome measures (PROs), (ii) to demonstrate and optimize sensitivity to change of many COAs, and (iii) to establish methods and evidence of anchoring change in COAs in patient meaningfulness, e.g., by determining patient-derived minimally meaningful thresholds of change. Keywords Activities of daily living (ADL) • Ataxia • Clinical outcome assessment (COA) • Scale for the assessment and rating of ataxia (SARA) • Standardization Ataxias, which have long been considered untreatable, are now becoming models for the development of targeted therapies. Consequently, an increasing number of observational studies and clinical trials is expected within the next years [1]. This situation requires harmonization and consensus on the use of appropriate clinical outcome assessments (COAs). In this manuscript, we provide (1) an overview of available COAs and (2) a proposal for the data content of ataxia registries based on the consensus of the Ataxia Global Initiative (AGI) working group on COAs.

Journal of the American College of Cardiology, Mar 1, 2002

As compared to patients of Group 1, those of Group 2 were younger (47±14 vs 41±15 years, p<0.001)... more As compared to patients of Group 1, those of Group 2 were younger (47±14 vs 41±15 years, p<0.001), and showed less advanced LV dysfunction (EF 28±9 vs 35±9%, p<0.001) and remodelling (end-diastolic diameter index 38±7 vs 35±7 minim2, p<0.001) at the time of diagnosis. Most patients of Group 1 and 2 were treated with ACE-I (93% and 88%, p=NS) and BB (83% and 74%, p<0.05). Five and 10-year transplant-free survival was respectively 73 and 57% in Group1 vs 93 and 86% in Group 2, while hospitatisation-free survival was 47 and 32% in Group1 vs 70 and 57% in Group 2 (both p<0.001). No outcome difference was observed between Group 2a and 2b, whereas 5 and 10-year transplant-free survival was significantly better in patients of Group 2 who were treated with BB than in those not receiving BB (p=0.007 after stratification for the severity of the disease). At 6 to 8 years of follow-up, 38% of patients of Group 2 developed HF symptoms, and/or a decrease of LVEF>10%, and/or the need of hospitalisation for cardiovascular reasons. The long-term progression of HF symptoms and LV dysfunction was similar in asymptomatic patients of Group 2a and 2b and not significantly different to that of patients of Group 1. Even though asymptomatic DC patients receiving an optimal medical treatment are characterized by low rates of death or heart transplant, nevertheless during a long-term follow-up they frequently exhibit a worsening of clinical status and LV function. Our data suggest that in these patients an early and aggressive BB strategy should be carefully considered in order to counteract as much as possible the Ioeg-term progression of the disease.

European Journal of Neuroscience, Jun 22, 2010

Pharmacological studies of narcoleptic canines indicate that exaggerated pontine cholinergic tran... more Pharmacological studies of narcoleptic canines indicate that exaggerated pontine cholinergic transmission promotes cataplexy. Since disruption of orexin (hypocretin) signaling is a primary defect in narcolepsy with cataplexy, we investigated whether markers of cholinergic synaptic transmission might be altered in mice constitutively lacking orexin receptors (double receptor knockout; DKO). We found that choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT) and the high-affinity choline transporter (CHT1) but not acetylcholinesterase (AChE) were significantly higher in samples from DKO compared to wild-type (WT) mice. This was region-specific since levels were elevated in samples from the laterodorsal tegmental nucleus (LDT) and the fifth motor nucleus (Mo5) but not in whole brainstem samples. Consistent with region-specific changes, we were unable to detect significant differences in Western blots for ChAT and CHT1 in isolates from brainstem, thalamus and cortex or in ChAT enzymatic activity in the pons. However, using ChAT immunocytochemistry, we found that while the number of cholinergic neurons in the LDT and Mo5 were not different, the intensity of somatic ChAT immunostaining was significantly greater in the LDT, but not Mo5, from DKOs compared to WTs. We also found that ChAT activity was significantly reduced in cortical samples from DKOs compared to WTs. Collectively, these findings suggest that the orexins can regulate neurotransmitter expression and that the constitutive absence of orexin signaling results in an upregulation of the machinery necessary for cholinergic neurotransmission in a mesopontine population of neurons that have been associated with both normal REM sleep and cataplexy.

PLOS ONE, Apr 13, 2011

To investigate how cholinergic systems regulate aspects of the sleep disorder narcolepsy, we vide... more To investigate how cholinergic systems regulate aspects of the sleep disorder narcolepsy, we video-monitored mice lacking both orexin (hypocretin) receptors (double knockout; DKO mice) while pharmacologically altering cholinergic transmission. Spontaneous behavioral arrests in DKO mice were highly similar to those reported in orexin-deficient mice and were never observed in wild-type (WT) mice. A survival analysis revealed that arrest lifetimes were exponentially distributed indicating that random, Markovian processes determine arrest lifetime. Low doses (0.01, 0.03 mg/kg, IP), but not a high dose (0.08 mg/ kg, IP) of the cholinesterase inhibitor physostigmine increased the number of arrests but did not alter arrest lifetimes. The muscarinic antagonist atropine (0.5 mg/kg, IP) decreased the number of arrests, also without altering arrest lifetimes. To determine if muscarinic transmission in pontine areas linked to REM sleep control also influences behavioral arrests, we microinjected neostigmine (50 nl, 62.5 mM) or neostigmine + atropine (62.5 mM and 111 mM respectively) into the nucleus pontis oralis and caudalis. Neostigmine increased the number of arrests in DKO mice without altering arrest lifetimes but did not provoke arrests in WT mice. Co-injection of atropine abolished this effect. Collectively, our findings establish that behavioral arrests in DKO mice are similar to those in orexin deficient mice and that arrests have exponentially distributed lifetimes. We also show, for the first time in a rodent narcolepsy model, that cholinergic systems can regulate arrest dynamics. Since perturbations of muscarinic transmission altered arrest frequency but not lifetime, our findings suggest cholinergic systems influence arrest initiation without influencing circuits that determine arrest duration.