Neuromuscular Disorders 33 (2023) S66-S192 treatment at 10mg/kg/day that was not observed at 20mg/kg/day.MitoQ-treated YS mice showed significantly decreased FDB fiber mitoSOX activity in response to stimulation (10 minutes) versus untreated and control mice.While elevated urine 15-F2t-isoprostane was not mitigated by ad lib MitoQ dosing, EM analyses revealed the absence or reduction of contracture/unstructured cores and z-line streaming.Completion of the combined dosing of MitoQ and n-3 PUFA will inform their efficacy on oxidative stress and muscle function in both mouse models.

Intracerebral hemorrhage (ICH) is a significant health concern associated with high mortality. Cofilin plays a crucial role in stress conditions, but its signaling following ICH in a longitudinal study is yet to be ascertained. In the present study, we examined the cofilin expression in human ICH autopsy brains. Then, the spatiotemporal cofilin signaling, microglia activation, and neurobehavioral outcomes were investigated in a mouse model of ICH. Human autopsy brain sections from ICH patients showed increased intracellular cofilin localization within microglia in the perihematomal area, possibly associated with microglial activation and morphological changes. Various cohorts of mice were subjected to intrastriatal collagenase injection and sacrificed at time points of 1, 3, 7, 14, 21, and 28 days. Mice suffered from severe neurobehavioral deficits after ICH, lasting for 7 days, followed by a gradual improvement. Mice suffered post-stroke cognitive impairment (PSCI) both acutely and in the chronic phase. Hematoma volume increased from day 1 to 3, whereas ventricle size increased from day 21 to 28. Cofilin protein expression increased in the ipsilateral striatum on days 1 and 3 and then decreased from days 7 to 28. An increase in activated microglia was observed around the hematoma on days 1 to 7, followed by a gradual reduction up to day 28. Around the hematoma, activated microglia showed morphological changes from ramified to amoeboid. mRNA levels of inflammatory [tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), and interleukin-6 (IL-6) and anti-inflammatory markers [interleukin-10 (IL-10), transforming growth factor-β TGF-β, and arginase I (Arg1)] increased during the acute phase and decreased in the chronic phase. Blood cofilin levels increased on day 3 and matched the increase in chemokine levels. slingshot protein phosphatase 1 (SSH1) protein, which activates cofilin, was increased from day 1 to 7. These results suggest that microglial activation might be the sequel of cofilin overactivation following ICH, leading to widespread neuroinflammation and consequent PSCI.

Intracerebral hemorrhage (ICH) is a major health concern associated with high mortality and morbidity. 50% of the stroke victims die within the first month and the rest of the survivors have to deal with long-term disabilities. The understanding of the molecular mechanism of the secondary injury-induced neuroinflammation or microglial activation is not studied well. Our previous study demonstrated a major causative role of the cytoskeletal protein, cofilin in ICH-induced brain injury. We demonstrated that knockdown of cofilin in a mouse model of collagenase induced-ICH improved neurobehavioral deficits, decreased hemorrhagic volume and microglia activation. In the present study, we aimed to study the cofilin signaling up to 14 days following ICH. We subjected different cohorts of mice to intrastriatal collagenase injection-induced ICH and mice were sacrificed at different time-points of 1, 3, 7, and 14 days. Using Western blotting (WB) and quantitative real-time PCR, we observed an upregulation of cofilin protein and mRNA levels in the ipsilateral striatum on day 3 and then a decreasing trend was observed from day 7 to day 14. Using immunohistochemistry analysis, activated microglia were observed to be increased after ICH from day 1, especially around the hematoma and lasted until day 7 and there was a gradual decrease observed at day 14. Activated microglia were associated with morphological changes from ramified into an amoeboid shape particularly around the hematoma from day 1 up to day 14. Human ICH autopsy brain sections also indicated that intracellular cofilin is localized within microglia and is associated with microglia morphological changes and activation of surrounding microglia. In conclusion, we believe that cofilin overactivation plays a causative role in the activation of microglia and subsequently leads to widespread neuroinflammation following ICH. Developing cofilin inhibitors might provide novel alternatives for ICH treatment.

Neuroinflammation, typified by elevated levels of interleukin-1 (IL-1) α and β, and deficits in proteostasis, characterized by accumulation of polyubiquitinated proteins and other aggregates, are associated with neurodegenerative disease independently and through interactions of the two phenomena. We investigated the influence of IL-1β on ubiquitination via its impact on activation of the E3 ligase parkin by either phosphorylated ubiquitin (P-Ub) or NEDD8. Immunohistochemistry and Proximity Ligation Assay were used to assess colocalization of parkin with P-tau or NEDD8 in hippocampus from Alzheimer patients (AD) and controls. IL-1β effects on PINK1, P-Ub, parkin, P-parkin, and GSK3β—as well as phosphorylation of parkin by GSK3β—were assessed in cell cultures by western immunoblot, using two inhibitors and siRNA knockdown to suppress GSK3β. Computer modeling characterized the binding and the effects of P-Ub and NEDD8 on parkin. IL-1α, IL-1β, and parkin gene expression was assessed by RT-PCR in brains of 2- and 17-month-old PD-APP mice and wild-type littermates. IL-1α, IL-1β, and parkin mRNA levels were higher in PD-APP mice compared with wild-type littermates, and IL-1α-laden glia surrounded parkin- and P-tau-laden neurons in human AD. Such neurons showed a nuclear-to-cytoplasmic translocation of NEDD8 that was mimicked in IL-1β-treated primary neuronal cultures. These cultures also showed higher parkin levels and GSK3β-induced parkin phosphorylation; PINK1 levels were suppressed. In silico simulation predicted that binding of either P-Ub or NEDD8 at a singular position on parkin opens the UBL domain, exposing Ser65 for parkin activation. The promotion of parkin- and NEDD8-mediated ubiquitination by IL-1β is consistent with an acute neuroprotective role. However, accumulations of P-tau and P-Ub and other elements of proteostasis, such as translocated NEDD8, in AD and in response to IL-1β suggest either over-stimulation or a proteostatic failure that may result from chronic IL-1β elevation, easily envisioned considering its early induction in Down’s syndrome and mild cognitive impairment. The findings further link autophagy and neuroinflammation, two important aspects of AD pathogenesis, which have previously been only loosely related.

We established a relationship between neuroinflammation, aggregate formation, and autophagic failure that occurs as a result of IL-1β-induced excess production of P-tau and Aβ aggregates, together with ApoE4-related failures in lysosomal autophagy; the latter giving rise to more plaques and tangles in APOE4,4 carriers. These ties between IL-1 and aggregate formation and clearance, as well as the known autophagy-related effects of the ubiquitin-like modifier NEDD8 on the E3-ligase parkin, prompted a new hypothesis explored here, viz., the profound neuroinflammation in AD is instrumental not only in aggregate formation, but also in aggregate processing through the NEDD8-parkin ubiquitination system. The subcellular distribution of parkin, NEDD8, and IL-1 was assessed immunochemically. NEDD8/parkin and parkin/P-tau was colocalized in hippocampal neurons in AD and AMC brains and in neural cell cultures by PLA. RT-PCR was used to assess their corresponding mRNAs in brains of young and old PD-APP mice. Subcellular fractionation and western blot were used to assess subcellular localization of NEDD8 in IL-1β-treated cells. Interactions between NEDD8 and parkin with E2-ubiquitin were predicted by computational simulations. Parkin-tau aggregates were more abundant in AD neurons adjacent to IL-1-expressing glia than were their counterparts in control patients, suggesting a potential connection between neuroinflammatory status and autophagy. Toward this, we found dual evidence of such a relationship: i) steady-state levels of IL-1α, IL-1β, and parkin mRNAs were elevated early in development in the PD-APP AD mouse model, and ii) IL-1β induced parkin expression in neuronal cell cultures. Further, NEDD8 was translocated from nucleus-to-cytoplasm in both primary cell cultures treated with IL-1β and in hippocampal neurons in Alzheimer brain; colocalization of parkin and NEDD8 was prominent in both. Computational modeling predicted NEDD8-parkin interactions, which may lead to increased affinity of parkin for E2-ubiquitin, the more active complex. Our findings are consistent with the idea that neuroinflammation results in dysfunctional interactions between the ubiquitin-like modifier NEDD8 and the E3-ligase parkin. The upregulation of parkin expression, alongside translocation of NEDD8 from nucleus-to-cytoplasm by excess IL-1β, suggests that such elevation fosters autophagy. However, this process may be thwarted by concomitant elevation of IL-1β-mediated P-tau aggregation.

Glioblastoma multiforme (GBM) is a World Health Organization (WHO) grade IV primary malignant astrocytoma. Aneurysms are devastating intracranial neurovascular pathologies. Intracranial dermoid cysts are common, benign lesions which can be clinically silent or associated with seizure disorder. We describe physically adjacent diagnoses of dermoid cyst, intracranial aneurysm, and GBM in a single patient. Records were collected and reviewed to compile the final clinical picture. A 72-year-old male with a long history of seizure disorder, presented with new focal, unilateral neurological deficits. Radiographic evaluation including computed tomography (CT) and magnetic resonance imaging (MRI) demonstrated a dermoid cyst with an underlying developing GBM, which also, by happenstance, contained an aneurysm. During open surgical resection, multiple macroscopically distinct tissue types were noted. Histological analysis of tissue from each lesion confirmed the diagnoses including dermoid cyst, GBM, and aneurysm. Pathological analysis revealed the presence of extensive inflammatory cells throughout. Subsequent staining identified CD68 positive cells indicating a probable chronic inflammatory state. Chronic inflammation resulting from the presence of a long term dermoid cyst and ongoing seizures may have led to dystrophic changes in adjacent vasculature and approximating glial tissues, inducing the formation of an aneurysm and a secondary GBM. Therefore, while benign in nature, dermoid cysts can be related to seizure disorder and may cause chronic inflammation in surrounding brain tissue.

Assessment of physician workloads has become increasingly important in modern academic physician practice, where it is commonly used to allocate resources among departments, to determine staffing, and to set the compensation of individual physicians. The physician work relative value unit system is a frequently used metric in this regard. However, the application of this system to the practice of pathology has proven problematic. One area of uncertainty is the validity of using work relative value unit norms that were derived from general surgical pathology practice to assess the various subspecialties within anatomic pathology. Here, we used data from the 2017 Association of Pathology Chairs practice survey to assess salary and work relative value unit data for single-subspecialty practitioners in US academic pathology departments in the prior year (2016). Five subspecialties were evaluated: dermatopathology, gastrointestinal pathology, hematopathology/hematology, renal pathology, and neuropathology. Data for general surgical pathologists and cytopathologists were included for comparison. For this analysis, survey data were available for 168 practitioners in 43 US academic departments of pathology. Salary ranges varied little among subspecialties, with the exception of dermatopathology, where salaries were higher. In contrast, work relative value unit productivity varied widely among different subspecialties, with median values differing as much as 4- to 7-fold between subspecialties. These results suggest that the use of a single overall work relative value unit standard is not appropriate for specialty- or subspecialty-based anatomic pathology practice, and that either the benchmark norms should be tailored to individual practice patterns, or an alternative system of workload measurement should be developed.

American hospitals are increasingly turning to service outsourcing to reduce costs, including laboratory services. Studies of this practice have largely focused on nonacademic medical centers. In contrast, academic medical centers have unique practice environments and unique mission considerations. We sought to elucidate and analyze clinical laboratory outsourcing experiences in US academic medical centers. Seventeen chairs of pathology with relevant experience were willing to participate in in-depth interviews about their experiences. Anticipated financial benefits from joint venture arrangements often eroded after the initial years of the agreement, due to increased test pricing, management fees, duplication of services in support of inpatients, and lack of incentive for utilization control on the part of the for-profit partner. Outsourcing can preclude development of lucrative outreach programs; such programs were successfully launched in several cases after joint ventures were either avoided or terminated. Common complaints included poor test turnaround time and problems with test quality (especially in molecular pathology, microbiology, and flow cytometry), leading to clinician dissatisfaction. Joint ventures adversely affected retention of academically oriented clinical pathology faculty, with adverse effects on research and education, which further exacerbated clinician dissatisfaction due to lack of available consultative expertise. Resident education in pathology and in other disciplines (especially infectious disease) suffered both from lack of on-site laboratory capabilities and from lack of teaching faculty. Most joint ventures were initiated with little or no input from pathology leadership, and input from pathology leadership was seen to have been critical in those cases where such arrangements were declined or terminated.

BACKGROUND: Cavernous hemangiomas account for 5% e13% of central nervous system vascular lesions. They are usually found intra-axially but rarely involve extra-axial structures, most commonly the middle cranial fossa. A cavernous hemangioma manifesting as a clinoid meningioma is extremely rare.CASE DESCRIPTION: A 36-year-old man presented with an asymptomatic intracranial mass found incidentally after a motor vehicle accident. Preoperative magnetic resonance imaging revealed a well-defined dural-based lesion arising from the right anterior clinoid process that was nearly homogeneously enhancing. The radiologic diagnosis was meningioma. The mass was resected via a right pterional craniotomy with microsurgical technique. Complete resection of the mass was performed with no complications and no significant bleeding. Contrasting with the radiologic and gross tumor appearance, histopathologic examination revealed dilated vascular spaces, sclerotic vessels without intervening neural tissue, and intravascular thrombi suggesting slow blood flow-all consistent with cavernous hemangioma.CONCLUSIONS: Anterior clinoid dural-based cavernous hemangiomas are extremely rare. Although preoperative diagnosis is difficult using imaging, this etiology should be considered for any dural-based middle fossa lesion, owing to the tendency for these lesions to bleed heavily during resection in some instances.

INTRODUCTION:Subependymomas are benign intraventricular tumours that most often occur asymptomatically and are found incidentally on autopsy. Symptomatic examples requiring surgical intervention are exceedingly rare.CASE PRESENTATION:A 55-year-old man with no history of neurological symptoms presented with multiple episodes of loss of consciousness and increasing headaches. MRI revealed a lobulated intraventricular mass centred at the right Foramen of Monro. Obstructive hydrocephalus with localised midline shift and a second lesion were noted. Right frontal craniotomy with total removal via transcortical resection was performed.DISCUSSION:Symptomatic subependymomas generally present with signs of hydrocephalus due to obstruction of cerebrospinal fluid pathways. There is only one other reported case of multifocal subependymomas in a symptomatic patient. An example of multiple supratentorial subependymomas causing obstructive hydrocephalus has not been previously reported.CONCLUSIONS:Multiple subependymomas are rare. Judicious surgical management with full excision led to symptomatic improvement in our patient.

INTRODUCTION:Alzheimer apolipoprotein E (APOE) ɛ4/ɛ4 carriers have earlier disease onset and more protein aggregates than patients with other APOE genotypes. Autophagy opposes aggregation, and important autophagy genes are coordinately regulated by transcription factor EB (TFEB) binding to "coordinated lysosomal expression and regulation" (CLEAR) DNA motifs.METHODS:Autophagic gene expression was assessed in brains of controls and Alzheimer's disease (AD) patients parsed by APOE genotype and in a glioblastoma cell line expressing either apoE3 or apoE4. Computational modeling assessed interactions between apoE and mutated apoE with CLEAR or modified DNA.RESULTS:Three TFEB-regulated mRNA transcripts-SQSTM, MAP1LC3B, and LAMP2-were lower in AD ɛ4/ɛ4 than in AD ɛ3/ɛ3 brains. Computational modeling predicted avid specific binding of apoE4 to CLEAR motifs. ApoE was found in cellular nuclei, and in vitro binding assays suggest competition between apoE4 and TFEB at CLEAR sites.CONCLUSION:ApoE4-CLEAR interactions may account for suppressed autophagy in APOE ɛ4/ɛ4 carriers and, in this way, contribute to earlier AD onset.

Introduction: Williams syndrome (WS), an autosomal dominant condition linked to gene deletions on chromosome 7, can cause supravalvular aortic narrowing and death. WS-associated mutations are believed to disrupt arterial elastin fibers, causing smooth muscle malformation, endomysial fibrosis and severe hypertension. Previous studies demonstrated arterial ultrastructural anomalies in adult WS patients. It is not presently known if the arterial phenotype of WS is also present in utero. Case report: A 34-week stillborn was delivered to a 28-year-old with genetically confirmed WS. Aortic tissue from the patient was compared with non-WS fetal aorta of similar gestational age using EM and light microscopy. Both sections were taken from the proximal aortic root. This demonstrated internal elastic lamina disruption, malformed elastic fibers, smooth muscle proliferation and abnormal collagen fibers, consistent with adult WS phenotype. Conclusion: Our analysis indicated the cardiovascular changes of WS in a fetus as young as 34 weeks.

Carotid splaying, also known as the Lyre sign, is a widening of the carotid bifurcation due to the displacement of the internal carotid artery and the external carotid artery just distal to the point of divergence. This phenomenon is classically exhibited by highly vascularized carotid body tumors and, in rare cases, by cervical sympathetic chain schwannomas. Demonstration of the Lyre sign by a cervical vagal neurofibroma, however, is a unique occurrence that has not been previously documented in the literature.Neurofibromas are slow growing, poorly vascularized soft tissue masses and are a hallmark of the autosomal dominant genetic disorder, neurofibromatosis type 1 (NF-1). While targeted genetic therapies are evolving, management is currently dependent on a case-by-case resection of tumors with specific indications for chemo and radiation therapy. These resections rely on magnetic resonance imaging (MRI) to visualize tumor location and infiltration, but even in the setting of an established NF-1 diagnosis, additional imaging can be beneficial in ruling out more precarious tumors and optimizing surgical outcomes.In this case, a 25-year-old female with known NF-1 presented with an enlarging cervical mass that demonstrated splaying of the left internal and external carotid arteries on MRI. Due to the typical association of the Lyre sign with carotid body tumors, magnetic resonance angiography (MRA) was crucial in guiding surgical decision making. Carotid body tumors are highly vascularized, may compress carotid branches, and carry a high risk of intraoperative bleeding. They are best visualized with MRA, which assesses carotid splaying and patency, and demonstrates vascular blushing within the tumor.This patient's MRA demonstrated the Lyre sign, patency of all carotid vessels, and a lack of vascularity within the mass, thus lowering suspicion for a carotid body tumor. Intraoperative use of imaging results facilitated a successful resection of a soft tissue tumor with minimal blood loss and no complications. Postoperative histologic examination confirmed a neurofibroma and definitively ruled out a carotid body tumor. This case highlights the importance of utilizing MRA whenever carotid splaying is seen on MRI and supports the consideration of neurofibromas in the differential for this finding.

Multiple spinal tumors are known to occur in association with phacomatosis like neurofibromatosis. Rarely, sporadic occurrences have been described in the absence of a clear underlying pathogenesis. The majority of intradural extramedullary spinal tumors are meningiomas or nerve sheath tumors like schwannoma and neurofibroma. Ependymomas, which are the most common intramedullary spinal tumors in adults, can also occur in the extramedullary intradural space and impose a diagnostic challenge. Only a few such cases have been described in the literature. We report the first sporadic occurrence of an intradural extramedullary ependymoma synchronously presenting with a different neoplasm, a schwannoma. A 43-year-old woman presented to the authors’ institution with lumbosacral pain and neurological deficits lasting a decade. On magnetic resonance imaging, she was found to have two stable, contrast-enhancing intradural extramedullary spinal lesions in the L2-3 and L4-5 vertebral areas. She subsequently underwent L2-L5 laminectomy with total resection of the former lesion and subtotal resection of the later lesion. Histological and immunohistochemical analysis revealed two distinct tumors: a cellular ependymoma and a schwannoma at the two respective locations. Ependymomas are low grade, slowly growing tumors that mimic other, benign tumors clinically as well as radiologically. However, the management of these tumors differs since ependymomas have a potential for recurrence and metastasis. The underlying etiology and pathogenesis of these tumors is not clearly established. However, an association with an underlying phacomatosis needs to be considered. Our purpose is to create awareness that an intradural extramedullary ependymoma can coexist with other spinal tumors and should be included in the differential diagnosis of a multifocal spinal lesion presentation. Owing to difficulties in differentiating these closely mimicking spinal tumors clinically and radiologically, early identification with surgical resection, when possible, might be a recommended strategy

The editors of Journal of Neuroinflammation would like to thank all our reviewers who have contributed to the journal in Volume 12 (2015).

Contributing reviewers The editors of Journal of Neuroinflammation would like to thank all our reviewers who have contributed to the journal in Volume 12 (2015).

Protoplasmic astrocytomas are a poorly characterized and extremely rare subtype of astrocytoma. We describe the CT, MR and 18F-fludeoxyglucose positron emission tomography (FDG-PET) findings of a multifocal protoplasmic astrocytoma in a 29-year-old male with neurological deficits. He was initially diagnosed with neurosarcoidosis based on imaging. MRI demonstrated intraparenchymal lesions involving the right temporal lobe and cerebellum. These appeared as extremely hyperintense signals on T2 weighted imaging and as homogeneous enhancements with a small non-enhancing cystic component on contrast-enhanced MR. Diffuse post-contrast enhancement of the craniospinal meninges was also noted. Post-radiation therapy PET-CT demonstrated a highly FDG-avid tumour in the right temporal lobe and left cerebellum. To our knowledge, a multifocal form of protoplasmic astrocytoma in an adult patient has not been previously described.