A small molecule toll-like receptor antagonist rescues α-synuclein fibril pathology.

The propagation and accumulation of pathological α-synuclein protein is thought to underlie the clinical symptoms of the neurodegenerative movement disorder Parkinson's disease (PD). Consequently, there is significant interest in identifying the mechanisms that contribute to α-synuclein pathology, as these may inform therapeutic targets for the treatment of PD. One protein that appears to contribute to α-synuclein pathology is the innate immune pathogen recognition receptor, toll-like receptor 2 (TLR2). TLR2 is expressed on neurons, and its activation results in the accumulation of α-synuclein protein; however, the precise mechanism by which TLR2 contributes to α-synuclein pathology is unclear. Herein we demonstrate using human cell models that neuronal TLR2 activation acutely impairs the autophagy lysosomal pathway and markedly potentiates α-synuclein pathology seeded with α-synuclein preformed fibrils. Moreover, α-synuclein pathology could be ameliorated with a novel small molecule TLR2 inhibitor, including in induced pluripotent stem cell-derived neurons from a patient with PD. These results provide further insight into how TLR2 activation may promote α-synuclein pathology in PD and support that TLR2 may be a potential therapeutic target for the treatment of PD.

Autophagy activation promotes clearance of α-synuclein inclusions in fibril-seeded human neural cells.

There is much interest in delineating the mechanisms by which the α-synuclein protein accumulates in brains of individuals with Parkinson's disease (PD). Preclinical studies with rodent and primate models have indicated that fibrillar forms of α-synuclein can initiate the propagation of endogenous α-synuclein pathology. However, the underlying mechanisms by which α-synuclein fibrils seed pathology remain unclear. To investigate this further, we have used exogenous fibrillar α-synuclein to seed endogenous α-synuclein pathology in human neuronal cell lines, including primary human neurons differentiated from induced pluripotent stem cells. Fluorescence microscopy and immunoblot analyses were used to monitor levels of α-synuclein and key autophagy/lysosomal proteins over time in the exogenous α-synuclein fibril-treated neurons. We observed that temporal changes in the accumulation of cytoplasmic α-synuclein inclusions were associated with changes in the key autophagy/lysosomal markers. Of note, chloroquine-mediated blockade of autophagy increased accumulation of α-synuclein inclusions, and rapamycin-induced activation of autophagy, or use of 5'-AMP-activated protein kinase (AMPK) agonists, promoted the clearance of fibril-mediated α-synuclein pathology. These results suggest a key role for autophagy in clearing fibrillar α-synuclein pathologies in human neuronal cells. We propose that our findings may help inform the development of human neural cell models for screening of potential therapeutic compounds for PD or for providing insight into the mechanisms of α-synuclein propagation. Our results further add to existing evidence that AMPK activation may be a therapeutic option for managing PD.

Nigrostriatal pathology with reduced astrocytes in LRRK2 S910/S935 phosphorylation deficient knockin mice.

Leucine-rich repeat kinase 2 (LRRK2) is genetically implicated in both familial and sporadic Parkinson's disease (PD). Moreover, LRRK2 has emerged as a compelling therapeutic target for the treatment of PD. Consequently, there is much interest in understanding LRRK2 and its role in PD pathogenesis. LRRK2 is constitutively phosphorylated on two serines, S910 and S935, that are required for interaction of LRRK2 with members of the 14-3-3 family of scaffolding proteins. Pathogenic LRRK2 missense mutations impair the phosphorylation of LRRK2 at these sites, but whether this contributes to PD pathology is unclear. To better understand how loss of LRRK2 phosphorylation relates to PD pathology, we have studied double knockin mice in which Lrrk2's serine 910 and 935 have both been mutated to alanine and can therefore no longer be phosphorylated. Nigrostriatal PD pathology was assessed in adult mice, aged mice, and mice inoculated with α-synuclein fibrils. Under all paradigms there was evidence of early PD pathology in the striatum of the knockin mice, namely alterations in dopamine regulating proteins and accumulation of α-synuclein. Striatal pathology was accompanied by a significant decrease in the number of astrocytes in the knockin mice. Despite striatal pathology, there was no degeneration of dopamine neurons in the substantia nigra and no evidence of a PD motor phenotype in the knockin mice. Our results suggest that modulation of LRRK2 serine 910 and 935 phosphorylation sites may have implications for dopamine turnover and astrocyte function, but loss of phosphorylation at these residues is not sufficient to induce PD neurodegeneration.

Toll-like receptor 2 is increased in neurons in Parkinson's disease brain and may contribute to alpha-synuclein pathology.

Inflammation is likely a key contributor to the pathogenesis of Parkinson's disease (PD), a progressively debilitating neurodegenerative disease that is accompanied by a pathological accumulation of the α-synuclein protein in a staged manner through the brain. What leads to the accumulation of α-synuclein in PD and how this relates to inflammatory pathways, however, is not entirely clear. Toll-like receptor (TLR) signaling is a major pathway mediating inflammation and, in particular, TLR2 is increasingly being implicated in PD. We have, therefore, examined the expression of TLR2 in postmortem brain tissue from PD patients and matched controls. We confirm that TLR2 is increased in PD brain, and find that levels of TLR2 correlate with the accumulation of pathological α-synuclein. TLR2 was expressed on neurons as well as microglia; however, the neuronal rather than glial expression of TLR2 was significantly increased in PD brain in accordance with disease staging, and TLR2 was strongly localized to α-synuclein positive Lewy bodies. In cell culture, activation of neuronal TLR2 induced an inflammatory response, including the secretion of inflammatory cytokines and microglial-activating chemokines, as well as the production of reactive oxygen species. Moreover, activation of neuronal TLR2 increased levels of endogenous α-synuclein protein, which was in turn associated with increased levels of the autophagy/lysosomal pathway marker p62. Finally, promoting autophagy with rapamycin or pharmacological inhibition of the TLR2 signaling pathway prevented the TLR2-mediated increase in α-synuclein in neuronal cell cultures. These results implicate neuronal TLR2 expression in human PD pathogenesis. In particular, the increased expression of TLR2 on neurons may provide new insight into disease pathogenesis and/or options for therapeutic intervention.

Impact of pilot diesel injection timing on performance and emission characteristics of marine natural gas/diesel dual-fuel engine.

In diesel-ignited natural gas marine dual-fuel engines, the pilot diesel injection timing (PDIT) determines the premixing time and ignition moment of the combustible mixture in the cylinder. The PDIT plays a crucial role in the subsequent development of natural gas flame combustion. In this paper, four PDITs (- 8 °CA, - 6 °CA, - 4 °CA, and - 2 °CA) were studied. The results show that the advancement of PDIT increased the engine's power, thermal efficiency, and natural gas flame spread velocity, and increased NO emissions and CH4 emissions of the marine engine. The PDIT affected the ignition delay period and the rapid combustion period to a greater extent than the slow combustion period and the post combustion period. With each 2 °CA advancement of PDIT, the engine's power increased by 69.87 kW, thermal efficiency increased by 0.42%, radial flame spread velocity increased by 2 m/s, axial flame spread velocity increased by 1.7 m/s, NO emissions increased by 6.1%, and CH4 emissions increased by 3.75%.

The impact of scavenging air state on the combustion and emission performance of marine two-stroke dual-fuel engine.

The scavenging process significantly affects the combustion and emission performance of marine low-speed two-stroke dual-fuel engines. Optimizing scavenging air pressure and temperature can enhance the engine's combustion efficiency and emission control performance, thereby achieving more environmentally friendly and efficient operation of dual-fuel engines. This study focuses on marine low-speed two-stroke dual-fuel engines, analyzing the effects of scavenging air pressure (3.0 bar, 3.25 bar, 3.5 bar, and 3.75 bar) and scavenging air temperature (293 K, 303 K, and 313 K) on engine performance and emission products. The results indicate that scavenging air pressure has a greater impact on engine performance than scavenging air temperature. An increase in scavenging air pressure leads to higher thermal efficiency and power. As the scavenging air pressure increases from 3 to 3.75 bar, the indicated thermal efficiency (ITE) increases from 44.02 to 53.26%, and indicated mean effective pressure (IMEP) increases by approximately 0.35 MPa. Increased scavenging air pressure improves nitrogen oxide (NOx) and hydrocarbons (HC) emissions. For every 0.25 bar increase in scavenging air pressure, NOx emissions decrease by 3.53%, HC emissions decrease by 33.35%, while carbon dioxide (CO2) emissions increase by 0.71%. An increase in scavenging air temperature leads to lower ITE and IMEP. As the air temperature changes from 293 to 313 K, the ITE decreases by approximately 1%, and IMEP decreases by about 0.04 MPa. Increased scavenging air temperature improves CO2 emissions. For every 10 K increase in the air temperature, the CO2 emissions decrease by 0.02%, while NOx emissions increase by 4.84%, HC emissions increase by 34.39%. Therefore, controlling scavenging air pressure is more important than scavenging air temperature in the operational management of marine two-stroke engines. Higher power and lower NOx and HC emissions can be achieved by increasing the scavenging air pressure.

Non-drug Therapies for Alzheimer's Disease: A Review.

Alzheimer's disease (AD) is a debilitating disease leading to great social and economic burdens worldwide. During the past decades, increasing understanding of this disease enables dynamic trials for disease interventions. Unfortunately, at present, AD still remains uncurable, and therefore, developing intervention strategies for improving symptoms and slowing down the disease process becomes a practical focus in parallel with searching for a disease-modifying medication. The aim of this review is to summarize the outcomes of AD clinical trials of non-drug therapies published in the past decade, including cognitive-oriented interventions, physical exercise interventions, brain stimulation, as well as nutrition supplementations, to find out the most effective interventions in the category by looking through the primary and secondary outcomes. The outcomes of the trials could be varied with the interventional approaches, the tested cohorts, the settings of observing outcomes, and the duration of follow-ups, which are all discussed in this review. Hence, we hope to provide crucial information for application of these interventions in real-world settings and assist with optimization of clinical trial designs in this area.

Effect of LRRK2 protein and activity on stimulated cytokines in human monocytes and macrophages.

Leucine-rich-repeat kinase 2 (LRRK2), a potential therapeutic target for the treatment of Parkinson's disease (PD), is highly expressed in monocytes and macrophages and may play a role in the regulation of inflammatory pathways. To determine how LRRK2 protein levels and/or its activity modulate inflammatory cytokine/chemokine levels in human immune cells, isogenic human induced pluripotent stem cells (iPSC) with the LRRK2-activating G2019S mutation, wild-type LRRK2, and iPSC deficient in LRRK2 were differentiated to monocytes and macrophages and stimulated with inflammatory toll-like receptor (TLR) agonists in the presence and absence of LRRK2 kinase inhibitors. The effect of LRRK2 inhibitors and the effect of increasing LRRK2 levels with interferon gamma on TLR-stimulated cytokines were also assessed in primary peripheral blood-derived monocytes. Monocytes and macrophages with the LRRK2 G2019S mutation had significantly higher levels of cytokines and chemokines in tissue culture media following stimulation with TLR agonists compared to isogenic controls. Knockout of LRRK2 impaired phagocytosis but did not significantly affect TLR-mediated cytokine levels. Interferon gamma significantly increased the levels of LRRK2 and phosphorylation of its downstream Rab10 substrate, and potentiated TLR-mediated cytokine levels. LRRK2 kinase inhibitors did not have a major effect on TLR-stimulated cytokine levels. Results suggest that the LRRK2 G2019S mutation may potentiate inflammation following activation of TLRs. However, this was not dependent on LRRK2 kinase activity. Indeed, LRRK2 kinase inhibitors had little effect on TLR-mediated inflammation under the conditions employed in this study.

Cognitive assessment tools for mild cognitive impairment screening.

Mild cognitive impairment (MCI) is a clinical condition with a high risk of progression to dementia. Due to lack of effective disease-modifying therapies for advanced dementia, diagnosis and disease intervention at an early stage, particularly at MCI stage, has been widely accepted as a critical strategy in disease management that could potentially affect long-term outcome. However, there is currently no consensus on guidelines for routine screening of MCI, resulting in a considerable number of patients with undiagnosed MCI from community. In addition, the use of different screening guidelines leads to difficulties in comparing different studies. A variety of screening tools have been utilized; however, the sensitivity and specificity vary greatly among these tools. By summarizing the sensitivity, specificity and time efficiency for common MCI screening tools, which are key factors to be taken into consideration when making selections and combinations of screening tools, this review suggests the use of a combination of two self-administered highly sensitive tools, p-AD8 + IQCODE (informant questionnaire on cognitive decline in the elderly individuals) in initial screening, as well as a combination of two highly specific widely covered tools, DemTect + MoCA (Montreal cognitive assessment) or memory and executive screening (MES) + MoCA in secondary screening. In addition, this review also proposes a screening flowchart for MCI, aiming to build a sensitive and time efficient way for recruiting subjects for subsequent investigation and disease differentiation.

The Development of Pharmacological Therapies for Alzheimer's Disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder that currently has no cure. The aged population is growing globally, creating an urgent need for more promising therapies for this debilitating disease. Much effort has been made in recent decades, and the field is highly dynamic, with numerous trials. The main focus of these trials includes disease modification and symptomatic treatment. Some have shown beneficial outcomes, while others have shown no significant benefits. Here, we cover the outcome of recently published AD clinical trials, as well as the mechanism of action of these therapeutical agents, to re-think drug development strategies and directions for future studies.

Can dipeptidyl peptidase-4 inhibitors treat cognitive disorders?

The linkage of neurodegenerative diseases with insulin resistance (IR) and type 2 diabetes mellitus (T2DM), including oxidative stress, mitochondrial dysfunction, excessive inflammatory responses and abnormal protein processing, and the correlation between cerebrovascular diseases and hyperglycemia has opened a new window for novel therapeutics for these cognitive disorders. Various antidiabetic agents have been studied for their potential treatment of cognitive disorders, among which the dipeptidyl peptidase-4 (DPP-4) inhibitors have been investigated more recently. So far, DPP-4 inhibitors have demonstrated neuroprotection and cognitive improvements in animal models, and cognitive benefits in diabetic patients with or without cognitive impairments. This review aims to summarize the potential mechanisms, advantages and limitations, and currently available evidence for developing DPP-4 inhibitors as a treatment of cognitive disorders.

Infectivity of human coronavirus in the brain.

A new strain of human coronaviruses (hCoVs), Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), has been identified to be responsible for the current outbreak of the coronavirus disease 2019 (COVID-19). Though major symptoms are primarily generated from the respiratory system, neurological symptoms are being reported in some of the confirmed cases, raising concerns of its potential for intracranial invasion and neurological manifestations, both in the acute phase and in the long-term. At present, it remains unclear the extent to which SARS-CoV-2 is present in the brain, and if so, its pathogenic role in the central nervous system (CNS). Evidence for neuroinvasion and neurovirulence of hCoVs has been recognised in animal and human studies. Given that SARS-CoV-2 belongs to the same family and shares characteristics in terms of receptor binding properties, it is worthwhile exploring its potential CNS manifestations. This review summarises previous findings from hCoVs in relation to the CNS, and compares these with the new strain, aiming to provide a better understanding of the effects of SARS-CoV-2 on the CNS.

Evaluation and accurate diagnoses of pediatric diseases using artificial intelligence.

Artificial intelligence (AI)-based methods have emerged as powerful tools to transform medical care. Although machine learning classifiers (MLCs) have already demonstrated strong performance in image-based diagnoses, analysis of diverse and massive electronic health record (EHR) data remains challenging. Here, we show that MLCs can query EHRs in a manner similar to the hypothetico-deductive reasoning used by physicians and unearth associations that previous statistical methods have not found. Our model applies an automated natural language processing system using deep learning techniques to extract clinically relevant information from EHRs. In total, 101.6 million data points from 1,362,559 pediatric patient visits presenting to a major referral center were analyzed to train and validate the framework. Our model demonstrates high diagnostic accuracy across multiple organ systems and is comparable to experienced pediatricians in diagnosing common childhood diseases. Our study provides a proof of concept for implementing an AI-based system as a means to aid physicians in tackling large amounts of data, augmenting diagnostic evaluations, and to provide clinical decision support in cases of diagnostic uncertainty or complexity. Although this impact may be most evident in areas where healthcare providers are in relative shortage, the benefits of such an AI system are likely to be universal.