PMCA Applications for Prion Detection in Peripheral Tissues of Patients with Variant Creutzfeldt-Jakob Disease

settings Order Article Reprints Open AccessReview PMCA Applications for Prion Detection in Peripheral Tissues of Patients with Variant Creutzfeldt-Jakob Disease by Giorgio Giaccone and Fabio Moda * [ORCID] Fondazione IRCCS Istituto Neurologico Carlo Besta, Division of Neurology 5—Neuropathology, 20133 Milan, Italy * Author to whom correspondence should be addressed. Biomolecules 2020, 10(3), 405; https://doi.org/10.3390/biom10030405 Submission received: 13 February 2020 / Revised: 2 March 2020 / Accepted: 5 March 2020 / Published: 5 March 2020 (This article belongs to the Special Issue Prion Disease Biomarkers: Recent Advances) Download keyboard_arrow_down Versions Notes Abstract Prion diseases are neurodegenerative and invariably fatal conditions that affect humans and animals. In particular, Creutzfeldt-Jakob disease (CJD) and bovine spongiform encephalopathy (BSE) are paradigmatic forms of human and animal prion diseases, respectively. Human exposure to BSE through contaminated food caused the appearance of the new variant form of CJD (vCJD). These diseases are caused by an abnormal prion protein named PrPSc (or prion), which accumulates in the brain and leads to the onset of the disease. Their definite diagnosis can be formulated only at post-mortem after biochemical and neuropathological identification of PrPSc. Thanks to the advent of an innovative technique named protein misfolding cyclic amplification (PMCA), traces of PrPSc, undetectable with the standard diagnostic techniques, were found in peripheral tissues of patients with vCJD, even at preclinical stages. The technology is currently being used in specialized laboratories and can be exploited for helping physicians in formulating an early and definite diagnosis of vCJD using peripheral tissues. However, this assay is currently unable to detect prions associated with the sporadic CJD (sCJD) forms, which are more frequent than vCJD. This review will focus on the most recent advances and applications of PMCA in the field of vCJD and other human prion disease diagnosis

Tackling amyloidogenesis in Alzheimer's disease with A2V variants of Amyloid-β

We developed a novel therapeutic strategy for Alzheimer’s disease (AD) exploiting the properties of a natural variant of Amyloid-β (Aβ) carrying the A2V substitution, which protects heterozygous carriers from AD by its ability to interact with wild-type Aβ, hindering conformational changes and assembly thereof. As prototypic compound we designed a six-mer mutated peptide (Aβ1-6A2V), linked to the HIV-related TAT protein, which is widely used for brain delivery and cell membrane penetration of drugs. The resulting molecule [Aβ1-6A2VTAT(D)] revealed strong anti-amyloidogenic effects in vitro and protected human neuroblastoma cells from Aβ toxicity. Preclinical studies in AD mouse models showed that short-term treatment with Aβ1-6A2VTAT(D) inhibits Aβ aggregation and cerebral amyloid deposition, but a long treatment schedule unexpectedly increases amyloid burden, although preventing cognitive deterioration. Our data support the view that the AβA2V-based strategy can be successfully used for the development of treatments for AD, as suggested by the natural protection against the disease in human A2V heterozygous carriers. The undesirable outcome of the prolonged treatment with Aβ1-6A2VTAT(D) was likely due to the TAT intrinsic attitude to increase Aβ production, avidly bind amyloid and boost its seeding activity, warning against the use of the TAT carrier in the design of AD therapeutics

Inhibitory Effects of oxalic acid on Listeria monocytogenes , Salmonella Enteritidis and Escherichia coli O157:H7 inoculated onto Chicken Breast stored at 4°C

Oxalic acid was evaluated for its effectiveness in inhibiting growth of selected pathogens on raw chicken breasts. Inoculated chicken breasts were dipped in oxalic acid solutions (0, 0.5, 1.0, 1.5 and 2.0% w/v) for 10, 20, and 30 min, packed in oxygenpermeable polyethylene bags, and stored at 4°C. Oxalic acid residues were determined using HPLC method. Counts of pathogens on chicken breasts were determined on days 0, 2, 5, 7, 10, and 14 after storage. Maximum oxalic acid concentration in unwashed chicken breast was 36 mg/100g. Washing of chicken reduced oxalic acid concentration by 50%. Oxalic acid concentration in cooked breast was 2mg/100g which is quite lower than levels in vegetables and herbs, used in daily diets. Chicken meat treated with oxalic acid could therefore be safe for human consumption. Reduction by 2.87, 2.02 and 4.12 log CFU/g, of L. monocytogenes, S. Enteritidis and E. coli O157:H7 respectively was observed in treated samples. Counts of the pathogens in treated samples decreased compared to untreated samples during 14 days storage. Sensory evaluation of cooked oxalic acid treated samples was acceptable to consumers after 14 days of storage. It was evident that oxalic acid has great potential for decontamination of chicken carcasses

Secondary Protein Aggregates in Neurodegenerative Diseases: Almost the Rule Rather than the Exception

The presence of protein aggregates is a hallmark of many neurodegenerative diseases, including Parkinson’s disease (PD), Alzheimer’s disease (AD), and frontotemporal lobar degeneration (FTLD). Traditionally, each disease has been associated with the aggregation of specific proteins, which serve as disease-specific biomarkers. For example, aggregates of α-synuclein (α-syn) are found in αsynucleinopathies such as PD, dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). Similarly, AD is characterized by aggregates of amyloid-beta (Aβ) and tau proteins. However, it has been observed that these protein aggregates can also occur in other neurodegenerative diseases, contributing to disease progression. For instance, α-syn aggregates have been detected in AD, Down syndrome, Huntington’s disease, prion diseases, and various forms of FTLD. Similarly, Aβ aggregates have been found in conditions like DLB and PD. Tau aggregates, in addition to being present in primary tauopathies, have been identified in prion diseases, α-synucleinopathies, and cognitively healthy aged subjects. Finally, aggregates of TDP-43, typically associated with FTLD and amyotrophic lateral sclerosis (ALS), have been observed in AD, progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), MSA, DLB, and other neurodegenerative diseases. These findings highlight the complexity of protein aggregation in neurodegeneration and suggest potential interactions and common mechanisms underlying different diseases. A deeper understating of this complex scenario may eventually lead to the identification of a better elucidation of the pathogenetic mechanisms of these devastating conditions and hopefully new therapeutic stragegies

Sporadic Creutzfeldt-Jakob disease: Real-Time Quaking Induced Conversion (RT-QuIC) assay represents a major diagnostic advance

Sporadic Creutzfeldt-Jakob disease (sCJD) is a rare and fatal neurodegenerative disorder with an incidence of 1.5 to 2 cases per million population/year. The disease is caused by a proteinaceous infectious agent, named prion (or PrPSc), which arises from the conformational conversion of the cellular prion protein (PrPC). Once formed, PrPSc interacts with the normally folded PrPC coercing it to undergo similar structural rearrangement. The disease is highly heterogeneous from a clinical and neuropathological point of view. The origin of this variability lies in the aberrant structures acquired by PrPSc. At least six different sCJD phenotypes have been described and each of them is thought to be caused by a peculiar PrPSc strain. Definitive sCJD diagnosis requires brain analysis with the aim of identifying intracerebral accumulation of PrPSc which currently represents the only reliable biomarker of the disease. Clinical diagnosis of sCJD is very challenging and is based on the combination of several clinical, instrumental and laboratory tests representing surrogate disease biomarkers. Thanks to the advent of the ultrasensitive Real-Time Quaking-Induced Conversion (RT-QuIC) assay, PrPSc was found in several peripheral tissues of sCJD patients, sometimes even before the clinical onset of the disease. This discovery represents an important step forward for the clinical diagnosis of sCJD. In this manuscript, we present an overview of the current applications and future perspectives of RT-QuIC in the field of sCJD diagnosis

Predictors of Bovine TB Risk Behaviour amongst Meat Handlers in Nigeria: A Cross-Sectional Study Guided by the Health Belief Model

Bovine Tuberculosis (bTB) is still a serious public health threat in developing countries. The aim of this study is to determine the social and cognitive factors predicting one of the risk behaviours amongst meat handlers in Nigeria, namely, eating Fuku Elegusi. This is the practice of eating the visibly infected parts of the lung in-order to convince customers to buy meat. The study is guided by the health belief model (HBM).This is a cross-sectional study of 349 randomly selected meat handlers in Oko-Oba Abattoir, in Lagos State. Descriptive statistics and multiple logistic regression analysis were employed to determine perceptions and prevalence of risk behaviours and to identify predictors of eating Fuku Elegusi.Just over a quarter (28.1%) of the study participants knew that eating Fuku Elegusi could be a source of bTB in humans. The prevalence of eating Fuku Elegusi was found to be 22%. Across all knowledge indicators related to bTB, those who don't eat Fuku Elegusi exhibited better knowledge. Strong predictors of eating Fuku Elegusi were: being male (OR: 2.39, 95% CI: 1.10 to 5.19; p = 0.03), not knowing that eating Fuku Elegusi exposes to bTB (OR: 3.72, 95% CI: 1.69 to 8.22; p = 0.001), and the perception that one cannot sell meat without tasting it (perceived barrier) (OR: 1.35, 95% CI: 1.13 to 1.60; p = 0.001). Lower risk of eating Fuku Elegusi was predicted by perceived susceptibility to bTB due to another risk behaviour, namely, not washing hands after handling meat (OR: 0.78, 95% CI: 0.64 to 0.96; p-value = 0.021). Television and radio were the most acceptable media for TB prevention messages (78.5% and 75.6% respectively).Meat handlers in developing countries bear high risk to bTB owing to prevailing social and cognition determinants. Findings were largely consistent with the propositions of HBM

Sporadic MM-1 Type Creutzfeldt-Jakob disease with hemiballic presentation and no cognitive impairment until death: How New NCJDRSU diagnostic criteria may allow early diagnosis

Sporadic Creutzfeldt-Jakob disease is the most common human prion disorder. Although associated with heterogeneous clinical phenotypes, its distinctive feature is the presence of a rapidly progressive multidomain cognitive impairment. We describe the atypical case of a patient affected by sporadic Methionine/Methionine type 1 Creutzfeldt-Jakob disease (typically associated with early cognitive decline) who presented with an isolated hemiballic syndrome and no signs of cognitive involvement until death. We review sporadic Creutzfeldt-Jakob disease diagnostic criteria and their updates since their first formulation, highlighting their limitations in clinical diagnostic work-up. Finally, we discuss the recently introduced National Creutzfeldt-Jakob Disease Research and Surveillance Unit diagnostic criteria, suggesting how their application could support an early clinical diagnosis, even in atypical cases, such as the one presented

The alpha-synuclein RT-QuIC products generated by the olfactory mucosa of patients with parkinson’s disease and multiple system atrophy induce inflammatory responses in SH-SY5Y cells

Parkinson’s disease (PD) and multiple system atrophy (MSA) are caused by two distinct strains of disease-associated α-synuclein (αSynD). Recently, we have shown that olfactory mucosa (OM) samples of patients with PD and MSA can seed the aggregation of recombinant α-synuclein by means of Real-Time Quaking-Induced Conversion (αSyn_RT-QuIC). Remarkably, the biochemical and morphological properties of the final α-synuclein aggregates significantly differed between PD and MSA seeded samples. Here, these aggregates were given to neuron-like differentiated SH-SY5Y cells and distinct inflammatory responses were observed. To deepen whether the morphological features of α-synuclein aggregates were responsible for this variable SH-SY5Y inflammatory response, we generated three biochemically and morphologically distinct α-synuclein aggregates starting from recombinant α-synuclein that were used to seed αSyn_RT-QuIC reaction; the final reaction products were used to stimulate SH-SY5Y cells. Our study showed that, in contrast to OM samples of PD and MSA patients, the artificial aggregates did not transfer their distinctive features to the αSyn_RT-QuIC products and the latter induced analogous inflammatory responses in cells. Thus, the natural composition of the αSynD strains but also other specific factors in OM tissue can substantially modulate the biochemical, morphological and inflammatory features of the αSyn_RT-QuIC products