bims-medebr 2025-06-29 papers

bims-medebr

Biomed News

on Metabolism of the developing brain

Issue of 2025–06–29
twelve papers selected by
Regina F. Fernández, Johns Hopkins University

Analysis of Energy Metabolism and Lipid Spatial Distribution in Hypoxic-Ischemic Encephalopathy Revealed by MALDI-MSI.
Astrocytic lipidopathy and bioenergetic failure in ApoE4-associated late-onset Alzheimer's disease: A unifying hypothesis.
Combined effects of β-hydroxybutyrate and therapeutic hypothermia in a neonatal hypoxia-ischemia model.
Lactate-induced metabolic signaling is the potential mechanism for reshaping the brain function - role of physical exercise.
The relationship between abnormal glucose metabolism and chronic pain.
Mass spectrometry-based spatiotemporal characterization of lipids in early focal cerebral ischemia.
Changes in glucose metabolism in essential tremor: within and beyond the cerebello-thalamo-cortical circuit.
Developmental Changes in Brain Cellular Membrane and Energy Metabolism: A Multi-Occasion 31P Magnetic Resonance Spectroscopy Study.
Lipidomic Network Analysis Reveals Amyloid-β-Induced Lysosomal Lipid Accumulation in the Cortex and Hippocampus of 5xFAD Mice.
Metabolomic alterations in human brain microvascular endothelial cells induced by traumatic injury.
The effects of hyperglycemia on brain physiology in a healthy and injured state: An experimental pig study with state-of-the-art multimodal neuromonitoring.
Uncovering injury-specific proteomic signatures and neurodegenerative risks in single and repetitive traumatic brain injury.

Biomedicines. 2025 Jun 11. pii: 1431. [Epub ahead of print]13(6):

Analysis of Energy Metabolism and Lipid Spatial Distribution in Hypoxic-Ischemic Encephalopathy Revealed by MALDI-MSI.

Xingxing Zhao, Peipei Chen, Lun Yu, Chuchu Gao, Sannan Wang, Zuming Yang, Zongtai Feng.

  Background: Neonatal hypoxic-ischemic encephalopathy (HIE) is a major cause of neonatal death and neurodevelopmental disorders, and its pathological mechanisms are closely related to disturbed energy metabolism and lipid remodeling. Exploring the spatial heterogeneity of metabolomics is essential to analyze the pathological process of HIE. Methods: In this study, we established a neonatal mouse hypoxic-ischemic brain damage (HIBD) model by the modified Rice method, and analyzed various metabolic pathways such as the tricarboxylic acid (TCA) cycle, purine metabolism, and lipid metabolism in the ischemic edema area, with contralateral and control brain tissues using matrix-assisted laser desorption mass spectrometry imaging (MALDI-MSI) with a spatial resolution of 50 μm. Results: In the HIBD model, key metabolites of the tricarboxylic acid (TCA) cycle (citrate, succinate, L-glutamate, glucose, aspartate, and glutamine) were significantly enriched in the edematous area compared with the control (fold change: 1.52-2.82), which suggests a blockage of mitochondrial function; ATP/ADP/AMP levels were reduced by 53-73% in the edematous area, and xanthine was abnormally accumulated in the hippocampus of the affected side, suggesting energy depletion and altered purine metabolism; lipid remodeling showed regional specificity: some unsaturated fatty acids, such as docosahexaenoic acid, were abnormally accumulated in the hippocampus. In contrast, pentadecanoic acid levels were reduced across the entire brain in the HIBD model, with a more pronounced decrease in the ipsilateral hippocampus, suggesting impaired membrane stability. Conclusions: The neonatal mouse HIBD model exhibits reprogramming of energy metabolism, characterized by a blockage in the tricarboxylic acid (TCA) cycle and ATP depletion, along with an abnormal spatial distribution of lipids. By targeting xanthine metabolic pathways, restoring mitochondrial function, and intervening in region-specific lipid remodeling, brain energy homeostasis may be improved and neurological damage attenuated. Further studies should validate the clinical feasibility of xanthine and lipid imbalance as diagnostic markers of HIBD and explore the critical time window for metabolic intervention to optimize therapeutic strategies.

Keywords:  energy metabolism; lipid metabolism; neonatal hypoxic-ischemic encephalopathy; purine metabolism; spatial metabolome

DOI:  https://doi.org/10.3390/biomedicines13061431
J Alzheimers Dis. 2025 Jun 26. 13872877251350338

Astrocytic lipidopathy and bioenergetic failure in ApoE4-associated late-onset Alzheimer's disease: A unifying hypothesis.

James P Garrahy.

  Late-onset Alzheimer's disease (LOAD) is traditionally attributed to amyloid-β (Aβ) accumulation and tau pathology as primary drivers of neurodegeneration. However, growing evidence suggests these may be secondary events arising from earlier disturbances in brain metabolism and lipid homeostasis. The ε4 allele of apolipoprotein E (ApoE4) remains the strongest genetic risk factor for LOAD, with carriers exhibiting both increased lifetime risk and earlier age of onset compared to ε2 or ε3 carriers. ApoE4 disrupts lipid metabolism and is associated with increased lipid droplet accumulation within astrocytes, implicating astrocytic lipidopathy in disease pathogenesis. Here, we propose a self-reinforcing pathogenic feedback loop-driven by dysregulated lipid homeostasis, chronic neuroinflammation, impaired glucose-handling, and cerebrovascular dysfunction-that culminates in astrocytic bioenergetic failure. This framework helps explain why ApoE4 carriers reach a critical bioenergetic threshold earlier in life, triggering the clinical onset of LOAD. Targeting astrocytic lipid homeostasis, through interventions such as blood-brain barrier-permeable statins, choline supplementation, or metabolic therapies, may offer novel strategies to delay disease progression or onset. Beyond AD, the framework proposed here, if validated, may have broader implications for unifying the cellular origins of age-related degenerative diseases and cancer through a shared vulnerability to progressive bioenergetic collapse.

Keywords:  Alzheimer's disease; apolipoprotein E; astrocytes; cerebral small vessel disease; cerebrovascular disease; insulin; lipid metabolism; lipidomics; lipids; neuroinflammation

DOI:  https://doi.org/10.1177/13872877251350338
J Cereb Blood Flow Metab. 2025 Jun 25. 271678X251352694

Combined effects of β-hydroxybutyrate and therapeutic hypothermia in a neonatal hypoxia-ischemia model.

Rafael Bandeira Fabres, Andrey Vinicios Soares Carvalho, Djuly Karoliny da Silva Alós, Diorlon Nunes Machado, Francielle Fernandes Spies, Débora Sterzeck Cardoso, Melany Ferreira Costa Vido, Ana Paula Rodrigues Martini, Marcel de Medeiros de Mattos, Bibiana Pereira Antunes, Eduarda Hoeper, Gianina Teribele Venturin, Samuel Greggio, Silvia Honda Takada, Alexander Drobyshevsky, Luciano Stürmer de Fraga, Carlos Alexandre Netto.

  Hypoxia-ischemia (HI) is one of the leading causes of brain damage during the development of newborns. It can result in death or cause varying degrees of neurological disability. The only well-established treatment currently available for neonatal HI is therapeutic hypothermia (TH). However, TH is only partially protective, reducing severe disability by approximately 11%. Therefore, new therapeutic approaches are urgently needed. It is known that immature brains utilize higher levels of ketone bodies, such as β-hydroxybutyrate (BHB), that may contribute to resistance to hypoxic-ischemic events. In this study, 11-day-old animals were subjected to the neonatal HI (Rice-Vannucci model) and treated with TH alone or in combination with BHB administration. To assess brain metabolism, glucose uptake was evaluated using MicroPET at 72 hours post-injury and when the animals reached 65 days of age. Behavioral tests, brain volume analysis, hippocampal cell counting and the assessment of hippocampal inflammatory cytokines expression were also performed. Animals treated with BHB exhibited increased glucose uptake at 72 hours post-injury and a reduction in neuronal loss in the hippocampus. The combined use of BHB and TH resulted in enhanced hippocampal neuronal survival, suggesting that BHB may represent a promising future treatment for neonatal HI.

Keywords:  Hypoxia-ischemia; MicroPET; brain metabolism; therapeutic hypothermia; β-hydroxybutyrate

DOI:  https://doi.org/10.1177/0271678X251352694
Front Endocrinol (Lausanne). 2025 ;16 1598419

Lactate-induced metabolic signaling is the potential mechanism for reshaping the brain function - role of physical exercise.

Xueqiang Zhu, Wenjia Chen, Ricardo A Pinho, Anand Thirupathi.

  Research into the effects of physical exercise on brain metabolism has revealed complex molecular mechanisms, with particular emphasis on lactate as a signaling molecule capable of transiently enhancing brain functions. This metabolite, once considered merely a byproduct of exercise, has been shown to enhance cognitive function through complex interactions with neural cells. This review examines how exercise-induced lactate formation acts as both an energy substrate and signaling molecule to reshape brain function, focusing on its metabolic and molecular mechanisms across different neural cell types. For that, we analyzed current literature on physical exercise-induced lactate production and its effects on brain metabolism, particularly examining lactate's dual role in cellular energetics and signaling pathways. The review synthesizes findings from both animal and human studies investigating exercise-dependent lactate mechanisms in brain function. We conducted a comprehensive analysis of peer-reviewed literature using databases including PubMed, Web of Science, and Scopus. The search terms included combinations of "physical exercise," "lactate," "brain metabolism," "cognitive function," and "neural plasticity." Both animal and human studies were included to provide a broad perspective on exercise-dependent lactate mechanisms in brain function. Understanding these lactate-mediated pathways is relevant for developing targeted physical exercise interventions that optimize brain health and cognitive function, potentially offering complementary therapeutic strategies for unfavorable neurological conditions.

Keywords:  brain health; energy homeostasis; lactate; metabolic signaling; physical exercise

DOI:  https://doi.org/10.3389/fendo.2025.1598419
Cell Biosci. 2025 Jun 21. 15(1): 86

The relationship between abnormal glucose metabolism and chronic pain.

Lulin Ma, Yaoling Wang, Yi Zhao, Meng Sun, Tao Zhu, Cheng Zhou.

  Chronic pain has a serious impact on the quality of patients' life. Currently, the mechanism of chronic pain has not been fully studied, and the treatments are often ineffective. Abnormal glucose metabolism plays an important role in the occurrence and development of chronic pain. It has been found that the abnormal glucose metabolism in anterior cingulate cortex (ACC), visual cortex, occipital cortex, brain stem, hippocampus, orbitofrontal cortex (OFC), thalamus and insula is involved in the primary headache (migraine); In addition, the abnormal glucose metabolism in the medial prefrontal cortex (mPFC), ACC, hippocampus, thalamus, primary somatosensory cortices (SI), OFC and cerebellum is involved in the neuropathic pain (NP); the abnormal glucose metabolism in the thalamus and brain stem is also involved in other types of chronic pain. Pain relieving therapies, such as transcranial magnetic stimulation (TMS), transcranial direct current stimulation (tDCS), electroacupuncture (EA) and acupuncture treatment can alleviate chronic pain by reversing abnormal glucose metabolism in some of the above brain regions. In conclusion, although further research is needed, the abnormal glucose metabolism and related treatment may be an important direction for the treatment of chronic pain, and relevant mechanisms still need to be further explored.

Keywords:  Abnormal glucose metabolism; Brain; Chronic pain; EA; Migraine; Neuropathic pain; TMS; tDCS

DOI:  https://doi.org/10.1186/s13578-025-01430-w
Anal Bioanal Chem. 2025 Jun 27.

Mass spectrometry-based spatiotemporal characterization of lipids in early focal cerebral ischemia.

Weifeng Li, Tengrui Shi, Zijun Liao, Yuan Zhang, Wenlan Liu.

  Understanding spatial lipid alterations in the brain during the hyper-acute phase of acute ischemic stroke (AIS) is crucial for elucidating the underlying mechanisms of disease progression and therapeutic response. In this study, we employed an integrated approach combining matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) and pseudo-targeted lipidomics to characterize lipid changes in the mice brain after 1 h, 2 h, and 3 h of middle cerebral artery occlusion (MCAO). Remarkably, MALDI-MSI detected significant lipid alterations in ischemic brain regions as early as 1 h post-MCAO, preceding visible tissue infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Lipidomics analysis further revealed region-specific lipid alterations at this early time point, suggesting that lipid profiles may serve as more sensitive biomarkers of ischemic brain injury than conventional histopathological methods. Notably, both MSI and lipidomics analysis demonstrated consistent temporal progression of lipid changes. Significantly altered lipids exhibited a nearly identical increasing trend with prolonged ischemic duration, with the ischemic cortex showing the most pronounced lipid dysregulation. In conclusion, our findings demonstrate that MALDI-MSI coupled with lipidomics provides a comprehensive platform for mapping the dynamic lipid landscape in the ischemic brain during early AIS, with observed patterns well correlating with the expansion of tissue infarction.

Keywords:  Acute ischemic stroke; Hyper-acute phase; Lipidomics; Mass spectrometry imaging

DOI:  https://doi.org/10.1007/s00216-025-05981-2
Brain Commun. 2025 ;7(3): fcaf227

Changes in glucose metabolism in essential tremor: within and beyond the cerebello-thalamo-cortical circuit.

Sarah Jonker, Jelle R Dalenberg, Debora E Peretti, A M Madelein van der Stouwe, Marina A J Tijssen.

  The pathophysiology and cerebellar role in essential tremor are not yet fully understood. Previous fludeoxyglucose positron emission tomography (FDG-PET) studies comparing glucose metabolism between essential tremor and healthy participants have led to few and inconsistent results. We aimed to examine changes in brain glucose metabolism in [18F]FDG PET brain imaging in 19 right-handed essential tremor patients and compare them to 19 right-handed healthy participants using a voxel-based mass univariate analysis. The Montreal Cognitive Assessment and the Hospital Depression scale were used as covariates. In addition, the correlation between tremor severity as measured with the Fahn-Tolosa-Marin Tremor Rating Scale and brain glucose metabolism in essential tremor patients was assessed. Essential tremor patients showed significantly higher metabolism in the right dentate nucleus and, at a more liberal threshold, lower metabolism in the right parietal cortex. A positive correlation was detected between glucose metabolism in the left paracentral region and tremor severity, again at more liberal thresholding. Beyond the cerebello-thalamo-cortical circuit, the decreased activity in the right parietal areas in essential tremor patients may indicate that the sensorimotor integration is an important aspect of essential tremor pathophysiology. In conclusion, our findings show altered glucose metabolism within and beyond the cerebello-thalamo-cortical circuit.

Keywords:  FDG-PET; cerebello-thalamo-cortical circuit; cerebellum; essential tremor

DOI:  https://doi.org/10.1093/braincomms/fcaf227
J Neurosci. 2025 Jun 26. pii: e2222242025. [Epub ahead of print]

Developmental Changes in Brain Cellular Membrane and Energy Metabolism: A Multi-Occasion 31P Magnetic Resonance Spectroscopy Study.

Yana Fandakova, Naftali Raz, Ulman Lindenberger, Dalal Khatib, Usha Rajan, Jeffrey A Stanley.

Structural neuroimaging studies of typical development reveal increases in grey matter volume during childhood, followed by shrinkage in adolescence and early adulthood. With neuropil constituting the bulk of grey matter, these developmental changes may reflect neuropil reorganization accompanied by alterations in cellular membranes, as well as changes in related energy demand. Phosphorus magnetic resonance spectroscopy (31P MRS) allows in vivo assessment of changes in the brain's high-energy phosphates - phosphocreatine (PCr), inorganic phosphate (Pi), and adenosine triphosphate (ATP) - as well as metabolites associated with synthesis and degradation of membrane phospholipids (MPLs) - phosphocholine (PC) and phosphoethanolamine (PE), and their breakdown products, glycerophosphocholine (GPC) and glycerophosphoethanolamine (GPE). Forty-nine children and adolescents aged 6 to 14 years at baseline (37 boys, 12 girls) were assessed on up to three occasions approximately 12 months apart. MPL precursor levels decreased across all examined regions over time, including cortical and subcortical gray matter and two major white matter tracts. Breakdown products increased in the prefrontal cortex (PFC) in younger children but decreased in their older counterparts. While ATP and Pi decreased across most regions, PCr changes were heterochronic and regional: Hippocampal increases were more pronounced in older children, whereas most of the remaining regions showed no change. Changes in MPL precursors were positively associated with change in PFC cortical thickness, suggesting that the expansion and contraction of neuropil are coupled with structural brain changes during childhood and adolescence. Thus, in vivo 31P MRS provides new insights into the neurobiological mechanisms of normal brain development.Significance Statement In childhood and adolescence, structural neuroimaging reveals marked changes in the brain's grey matter, most likely indicating contraction and expansion of its main component - the neuropil. The neurobiological mechanisms of these changes are, however, poorly understood. In the first of its kind longitudinal study of 6- to 14-year-old children, we examined in vivo changes in metabolites associated with brain energetics and the synthesis and degradation of membrane phospholipids using phosphorus magnetic resonance spectroscopy. We identify developmental changes in the metabolites associated with contraction and expansion of the neuropil and their coupling with structural changes in late-to-mature brain regions of the prefrontal cortex, indicating candidate mechanisms of brain development.

DOI: https://doi.org/10.1523/JNEUROSCI.2222-24.2025
J Proteome Res. 2025 Jun 24.

Lipidomic Network Analysis Reveals Amyloid-β-Induced Lysosomal Lipid Accumulation in the Cortex and Hippocampus of 5xFAD Mice.

Yun Jae Cha, Young-Kwang Kim, Yun Ji Lim, Chan Ho Kim, Se Eun Park, Yun Pyo Kang, Min-Kyoo Shin.

  The 5xFAD mouse model serves as a valuable experimental system for investigating Alzheimer's disease (AD), specifically amyloid-beta (Aβ)-induced AD pathology. In this study, we explored temporal, regional, and sex-specific alternations in the lipidome within the cortex and hippocampus of 5xFAD mice. Our results revealed that lipid alternations become more pronounced with the progression of Aβ pathology in the cerebral cortex and hippocampus. These lipid changes were also more significant in the female mice, which exhibited more severe Aβ pathology than male mice. Through lipid network analysis, we identified AD-specific lipid coexpression network modules in both brain regions, marked by enriched lysosomal lipids such as BMP and GM3. Notably, this lipid profile was also observed in microglia cells overexpressing the Swedish mutant form of Aβ precursor protein (APPswe). Given the critical role of BMP in lysosomal lipid and membrane degradation, and the observed enhancement of GM3 accumulation under lysosomal inhibition in APPswe-transfected microglial cells, these findings suggest that Aβ-mediated microglial lysosomal dysfunction may contribute to AD progression. Overall, we discovered a previously unrecognized role of Aβ in dysregulating lysosomal lipid metabolism and highlighted the utility of lipidomics and network analysis as complementary approaches for elucidating disease mechanisms.

Keywords:  5xFAD; Alzheimer’s disease; WGCNA; lipidomics; lysosome

DOI:  https://doi.org/10.1021/acs.jproteome.4c01133
Metabolomics. 2025 Jun 27. 21(4): 93

Metabolomic alterations in human brain microvascular endothelial cells induced by traumatic injury.

Enis Cela, David Tweddell, Eric K Patterson, Mark Daley, Gediminas Cepinskas, Douglas D Fraser.

   INTRODUCTION: Altered metabolic pathways are critical in the progression of traumatic brain injury (TBI). Identifying differentially abundant metabolites (DAMs) from specific cell types can offer valuable diagnostic and prognostic insights.
OBJECTIVE: This study aimed to characterize the metabolomic profile of injured human brain microvascular endothelial cells (hBMEC) at 2-, 12-, 24-, and 48 h post-injury.
METHODS: Using an in vitro TBI model, we analyzed metabolites in cell culture media through a combination of direct injection mass spectrometry and a custom reverse-phase LC-MS/MS assay. We evaluated 644 metabolites at each time point.
RESULTS: Phosphatidylcholines were significantly upregulated across all time intervals. At 2- and 12 h post-injury, the most significantly upregulated metabolites included sphingomyelin (OH) C22:1, ethylmalonic acid, and methylhistidine, while guanosine and the combination of butyric acid + isobutyric acid were the most downregulated. At 24 and 48 h, deoxyadenosine and inosine, respectively, emerged as the most upregulated metabolites, with butyric acid + isobutyric acid and quinoline-4-carboxylic acid showing the greatest downregulation.
CONCLUSION: Metabolomic profiling identified various DAMs after traumatic injury that are linked to human endothelial dysfunction. Future experiments should expand the number of metabolites measured to determine the underlying signaling pathways.

Keywords:  Culture; Endothelial cells; Media; Metabolites; Traumatic brain injury

DOI:  https://doi.org/10.1007/s11306-025-02286-4
J Cereb Blood Flow Metab. 2025 Jun 26. 271678X251337633

The effects of hyperglycemia on brain physiology in a healthy and injured state: An experimental pig study with state-of-the-art multimodal neuromonitoring.

Teodor Svedung Wettervik, Anders Hånell, Kerstin M Ahlgren, Henrik Engquist, Anders Lewén.

  Although hyperglycemia is associated with worse outcome following acute brain injury, the pathomechanisms remain elusive. In this experimental pig study, we explored the effects of hyperglycemia on brain physiology. Six pigs were anesthetized and received multimodal neuromonitoring of intracranial pressure (ICP), cerebral perfusion pressure (CPP), cerebral autoregulatory metrics (PRx, CBFx, ORx, and with transfer function analysis), cerebral blood flow (CBF), partial brain tissue oxygenation (pbtO2), and cerebral microdialysis (MD). The effect of hyperglycemia was studied in the healthy brain after injection of intravenous glucose injections, which increased MD-glucose, while ICP, CPP, PRx, CBFx, ORx pbtO2, and cerebral energy metabolism remained unchanged. After normalization of arterial glucose, an intracranial balloon was inflated to increase ICP, followed by an intravenous glucose injection to study the effect of hyperglycemia in the injured brain. The latter induced a significant CBF elevation, but no changes in PRx, pbtO2, or cerebral energy metabolism (but a trend towards higher glucose). Hyperglycemia led to favorable short-term effects on cerebral physiology and the immediate increase in arterial glucose that usually follows acute brain injury may be physiologically neuroprotective and the detrimental role of hyperglycemia is more likely related to cellular and molecular pathophysiological mechanisms or merely a confounder.

Keywords:  Brain injury; brain physiology; cerebral blood flow; hyperglycemia; inverse translational; neurointensive care

DOI:  https://doi.org/10.1177/0271678X251337633
Signal Transduct Target Ther. 2025 Jun 23. 10(1): 195

Uncovering injury-specific proteomic signatures and neurodegenerative risks in single and repetitive traumatic brain injury.

Sarah Mantash, Soulaimane Aboulouard, Hassan Dakik, Yanis Zirem, Lydia Ziane-Chaouche, Ali Nehme, Khalil Mallah, Marya El-Kurdi, Naify Ramadan, Isabelle Fournier, Kazem Zibara, Firas Kobeissy, Michel Salzet.

Traumatic brain injury (TBI) is a major public health concern associated with an increased risk of neurodegenerative diseases including Alzheimer's disease (AD), Parkinson's disease (PD), and chronic traumatic encephalopathy, yet the underlying molecular mechanisms in repetitive TBI remain poorly defined. This study investigates proteomic and behavioral changes following single and repetitive mild TBI in a mouse model, focusing on molecular alterations in the cortex and hippocampus across acute (48 h) and subacute (1 week) stages. Using shotgun proteomics and bioinformatics approaches, including weighted gene co-expression network analysis (WGCNA) and machine learning, we analyzed the proteomic landscapes of TBI-affected brain regions including the hippocampus and the cortex. We assessed motor and cognitive outcomes at 2-, 7-, and 30-days post-injury to explore functional impairments associated with observed molecular changes. Our findings reveal spatio-temporal injury- and time-specific proteomic changes, with a single TBI promoting neuroprotective and repair mechanisms, while repetitive TBI exacerbating neuronal damage and synaptic deficits in the hippocampus. Key deregulated proteins, including Apoa1, ApoE, Cox6a1, and Snca, were linked to neurodegenerative pathways, suggesting molecular connections between TBI and diseases like AD and PD. Behavioral assessments indicated that repetitive TBI significantly impaired motor and cognitive functions, with recovery in motor function by day 30, whereas cognitive deficits persisted. This study provides a detailed analysis of the proteomic and behavioral consequences of TBI, identifying molecular networks as potential biomarkers or therapeutic targets for mitigating long-term cognitive decline associated with repetitive head trauma. These findings underscore the importance of mitochondrial and synaptic integrity in TBI response and suggest that targeting these pathways could reduce neurodegenerative risk following repetitive TBI.

DOI: https://doi.org/10.1038/s41392-025-02286-9