Lipopolysaccharide, Identified Using an Antibody and by PAS Staining, Is Associated With Corpora amylacea and White Matter Injury in Alzheimer's Disease and Aging Brain

Corpora amylacea (CA) increase in number and size with aging. Their origins and functions remain unknown. Previously, we found that Alzheimer's disease (AD) brains have more CA in the periventricular white matter (PVWM) compared to aging controls. In addition, CA is associated with neurodegeneration as indicated by colocalization of degraded myelin basic protein (dMBP) with periodic acid-Schiff (PAS), a CA marker. We also found that bacterial lipopolysaccharide is present in aging brains, with more LPS in AD compared with controls. Periodic acid-Schiff staining is used to identify CA by virtue of their high polysaccharide content. Despite the growing knowledge of CA as a contributor to AD pathology, the molecules that contribute to the polysaccharides in CA are not known. Notably, lipopolysaccharides (LPS) are important cell-surface polysaccharides found in all Gram-negative bacteria. However, it is unknown whether PAS could detect LPS, whether the LPS found in aging brains contribute to the polysaccharide found in CA, and whether LPS associate with myelin injury. In this study, we found that aging brains had a myelin deficit zone (MDZ) adjacent to the ventricles in PVWM. The MDZ contained vesicles, most of which were CA. LPS and dMBP levels were higher in AD than in control brains. LPS was colocalized with dMBP in the vesicles/CA, linking white matter injury with a bacterial pro-inflammatory molecule. The vesicles also contained oxidized fibers, C-reactive protein, NG2, and GALC, markers of oligodendrocyte precursor cells (OPCs) and oligodendrocyte cells (OLs), respectively. The vesicles/CA were surrounded by dense astrocyte processes in control and AD brains. LPS was co-localized with CA by double staining of PAS with LPS in aging brains. The relationship of LPS with PAS staining was confirmed by PAS staining of purified LPS on nitrocellulose membranes. These findings reveal that LPS is one of the polysaccharides found in CA which can be stained with PAS. In addition, vesicles/CA are associated with oxidized and damaged myelin. The LPS in these vesicles/CA may have contributed to this oxidative myelin damage and may have contributed to oxidative stress to OPCs and OLs which could impair the ability to repair damaged myelin in AD and control brains.

Quantitative analysis of size and regional distribution of corpora amylacea in the hippocampal formation of obstructive sleep apnoea patients

Corpora amylacea (CoA) are spherical aggregates of glucose polymers and proteins within the periventricular, perivascular and subpial regions of the cerebral cortex and the hippocampal cornu ammonis (CA) subfields. The present study quantified the distribution of CoA in autopsied hippocampi of patients with obstructive sleep apnoea (OSA) using ethanolamine-induced fluorescence. CoA were observed in 29 of 30 patients (96.7%). They were most abundant in periventricular regions (wall of lateral ventricle, alveus, fimbria and CA4), rarely found in the CA3 and CA1, and undetectable in the CA2 or subiculum. A spatiotemporal sequence of CoA deposition was postulated, beginning in the fimbria and progressively spreading around the subpial layer until they extended medially to the wall of the lateral ventricle and laterally to the collateral sulcus. This ranked CoA sequence was positively correlated with CoA packing density (count and area fraction) and negatively correlated with CoA minimum diameters (p < 0.05). Although this sequence was not correlated with age or body mass index (BMI), age was positively correlated with the mean and maximum diameters of CoA. These findings support the view that the spatiotemporal sequence of CoA deposition is independent of age, and that CoA become larger due to the accretion of new material over time.

Corpora Amylacea in the Human Brain Exhibit Neoepitopes of a Carbohydrate Nature

Corpora amylacea (CA) in the human brain are polyglucosan bodies that accumulate residual substances originated from aging and both neurodegenerative and infectious processes. These structures, which act as waste containers, are released from the brain to the cerebrospinal fluid, reach the cervical lymph nodes via the meningeal lymphatic system and may be phagocytosed by macrophages. Recent studies indicate that CA present certain neoepitopes (NEs) that can be recognized by natural antibodies of the IgM class, and although evidence of different kinds suggests that these NEs may be formed by carbohydrate structures, their precise nature is unknown. Here, we adapted standard techniques to examine this question. We observed that the preadsorption of IgMs with specific carbohydrates has inhibitory effects on the interaction between IgMs and CA, and found that the digestion of CA proteins had no effect on this interaction. These findings point to the carbohydrate nature of the NEs located in CA. Moreover, the present study indicates that, in vitro, the binding between certain natural IgMs and certain epitopes may be disrupted by certain monosaccharides. We wonder, therefore, whether these inhibitions may also occur in vivo. Further studies should now be carried out to assess the possible in vivo effect of glycemia on the reactivity of natural IgMs and, by extension, on natural immunity.

Brainstem neuropathology in two cases of COVID-19: SARS-CoV-2 trafficking between brain and lung

Introduction SARS-CoV-2 might spread through the nervous system, reaching respiratory centers in the brainstem. Because we recently reported neurophysiological brainstem reflex abnormalities in COVID-19 patients, we here neuropathologically assessed structural brainstem damage in two COVID-19 patients. Materials and methods We assessed neuropathological features in two patients who died of COVID-19 and in two COVID-19 negative patients as controls. Neuronal damage and corpora amylacea (CA) numbers /mm2 were histopathologically assessed. Other features studied were the immunohistochemical expression of the SARS-CoV-2 nucleoprotein (NP) and the Iba-1 antigen for glial activation. Results Autopsies showed normal gross brainstem anatomy. Histopathological examination demonstrated increased neuronal and CA damage in Covid-19 patients’ medulla oblongata. Immunohistochemistry disclosed SARS-CoV-2 NP in brainstem neurons and glial cells, and in cranial nerves. Glial elements also exhibited a widespread increase in Iba-1 expression. Sars-Co-V2 was immunohistochemically detected in the vagus nerve fibers. Discussion Neuropathologic evidence showing SARS-CoV-2 in the brainstem and medullary damage in the area of respiratory centers strongly suggests that the pathophysiology of COVID-19-related respiratory failure includes a neurogenic component. Sars-Co-V2 detection in the vagus nerve, argues for viral trafficking between brainstem and lung.

Brain Lesions in Aging Zoo-Housed Naked Mole-Rats (Heterocephalus glaber)

Naked mole-rats (NMRs) are common in the managed care of zoos and valuable models for aging research. Limited information on NMR neuropathology is available despite many studies regarding their aging physiology. Histologic sections of brain from 27 adult (5–27 years old) NMRs from 2 zoos were reviewed to determine presence or absence of lesions associated with advanced age in humans and other mammals. A majority (23/27; 85%) of NMR brains had cerebral cortical neuronal changes with rounded or angular neurons, cytoplasmic vacuoles containing pale yellow pigment, periodic acid–Schiff (PAS)-positive granules and green autofluorescence, compatible with lipofuscinosis. Less severe lesions were present in cerebellar Purkinje cells, medulla, and hippocampal neurons. The hypothalamic neuropil of all NMRs had scattered variably sized PAS-positive granules and 10 (37%) had larger round bodies consistent with corpora amylacea. The youngest NMRs, 5 to 7 years old, generally had minimal or no cerebrocortical lesions. Further studies will help understand brain aging in this long-lived species.

Evaluation of corpora amylacea in refractory epilepsy

Objectives: The purpose of this study is to check whether there is or not a relationship between CoA density and different clinical parameters of the disease established. Compare the assessment of CoA with Hematoxylin-Eosin technique against PAS staining. It is also intended to check if it exists a relationship between hippocampal CoA density and Nestine overexpression. Methods: Histological analysis of 14 patients. For the counting of the CoA, raw scores were used interchangeably, as well as discretized values on a semi-quantitative scale according to Cherian et al. criteria. The modified Engel scale was used to measure post-surgical evolution. Results: The proportion of CoA in the hippocampus matches the proportion found in different studies: 64.3%. At α=0.05, there is significant evidence to observe that the counting is different depending on the type of staining used in the slides. The average number of CoA is higher with the PAS. It is seen that the density of CoA in the hippocampus and cortex is directly related to its density in parahypocampal structures. With regard to the relation between CoA density and age of onset, there are signs of significance. As well as in the post-surgical evolution. With regard to the duration of the crisis period, was found that both variables are independent. Finally, it is observed that there is a direct association between Nestine overexpression and hippocampal CoA density. Conclusion: In patients with Refractory Epilepsy (ER), CoA presence is commonly verified, with a better evaluation with PAS staining. Respectfully to the relation between CoA density and age of onset, there are signs of significance, as well as post-surgical evolution with the modified Engel scale. It would be convenient to continue with the investigations in this field to contrast the role of CoA in ER.