Amyloid-ß and Phosphorylated Tau are the Key Biomarkers and Predictors of Alzheimer's Disease.

Alzheimer's disease (AD) is a age-related neurodegenerative disease and is a major public health concern both in Texas, US and Worldwide. This neurodegenerative disease is mainly characterized by amyloid-beta (Aß) and phosphorylated Tau (p-Tau) accumulation in the brains of patients with AD and increasing evidence suggests that these are key biomarkers in AD. Both Aß and p-tau can be detected through various imaging techniques (such as positron emission tomography, PET) and cerebrospinal fluid (CSF) analysis. The presence of these biomarkers in individuals, who are asymptomatic or have mild cognitive impairment can indicate an increased risk of developing AD in the future. Furthermore, the combination of Aß and p-tau biomarkers is often used for more accurate diagnosis and prediction of AD progression. Along with AD being a neurodegenerative disease, it is associated with other chronic conditions such as cardiovascular disease, obesity, depression, and diabetes because studies have shown that these comorbid conditions make people more vulnerable to AD. In the first part of this review, we discuss that biofluid-based biomarkers such as Aß, p-Tau in cerebrospinal fluid (CSF) and Aß & p-Tau in plasma could be used as an alternative sensitive technique to diagnose AD. In the second part, we discuss the underlying molecular mechanisms of chronic conditions linked with AD and how they affect the patients in clinical care.

Correction: Reddy et al. Rlip Reduction Induces Oxidative Stress and Mitochondrial Dysfunction in Mutant Tau-Expressed Immortalized Hippocampal Neurons: Mechanistic Insights. Cells 2023, 12, 1646.

The authors wish to make the following changes to their paper [...].

Methods to Study Mitochondria: Techniques Used to Study the Effects of Age-Related Diseases Including Alzheimer's.

Various laboratories across the world have developed methods to study mitochondrial proteins/markers through extraction of mitochondrial RNA and protein to assess mitophagy/autophagy in Alzheimer's disease and other age-related diseases. Techniques outlined in this article include qRT-PCR, immunoblotting, immunofluorescence, transmission electron microscopy, Seahorse bioanalysis, staining for mitochondrial membrane potential, detection of mitophagy, and mitochondrial functional assays. Most of these techniques have been performed in vitro (in human and mouse neuronal cell lines transfected with mutant amyloid precursor protein or tau protein cDNAs) and in vivo (in brain tissues from different mouse models of Alzheimer's and other neurological diseases). Mitochondrial abnormalities in Alzheimer's disease have taken various forms, including excessive reactive oxygen species production, mitochondrial calcium dyshomeostasis, loss of ATP, defects in mitochondrial dynamics and transport, and mitophagy. Mitochondrial dysfunction is largely involved in aging; age-related diseases such as cancer, diabetes, and obesity; and neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, and others. The goal of this article is to make protocols/methods available to students, scholars, and researchers of mitochondria in order to facilitate future mitochondrial studies. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Analyzing mitochondrial gene expression in mouse brain tissue and HT22 cells by qRT-PCR Basic Protocol 2: Analyzing protein expression in mouse brain tissue and HT22 cells by immunoblotting Basic Protocol 3: Immunofluorescence staining of cells and tissue sections Basic Protocol 4: Staining for mitochondrial membrane potential Basic Protocol 5: Assessing mitochondrial structure by transmission electron microscopy Basic Protocol 6: Methods for detecting mitophagy Basic Protocol 7: Bioenergetics assay via Seahorse Basic Protocol 8: Assays for mitochondrial function.

Mitochondria in Aging and Alzheimer's Disease: Focus on Mitophagy.

Alzheimer's disease (AD) is characterized by the accumulation of amyloid ß and phosphorylated τ protein aggregates in the brain, which leads to the loss of neurons. Under the microscope, the function of mitochondria is uniquely primed to play a pivotal role in neuronal cell survival, energy metabolism, and cell death. Research studies indicate that mitochondrial dysfunction, excessive oxidative damage, and defective mitophagy in neurons are early indicators of AD. This review article summarizes the latest development of mitochondria in AD: 1) disease mechanism pathways, 2) the importance of mitochondria in neuronal functions, 3) metabolic pathways and functions, 4) the link between mitochondrial dysfunction and mitophagy mechanisms in AD, and 5) the development of potential mitochondrial-targeted therapeutics and interventions to treat patients with AD.

Rlip Reduction Induces Oxidative Stress and Mitochondrial Dysfunction in Mutant Tau-Expressed Immortalized Hippocampal Neurons: Mechanistic Insights.

RalBP1 (Rlip) is a stress-activated protein that is believed to play a large role in aging and neurodegenerative diseases such as Alzheimer's disease (AD) and other tauopathies. The purpose of our study was to understand the role of Rlip in mutant Tau-expressed immortalized hippocampal HT22 cells. In the current study, we used mutant Tau (mTau)-expressed HT22 neurons and HT22 cells transfected with Rlip-cDNA and/or silenced RNA, and studied the cell survival, mitochondrial respiration, mitochondrial function, immunoblotting, and immunofluorescence analysis of synaptic and mitophagy proteins and the colocalization of Rlip and mTau proteins. We found Rlip protein levels were reduced in mTau-HT22 cells, Rlip silenced HT22 cells, and mTau + Rlip RNA silenced HT22 cells; on the other hand, increased Rlip levels were observed in Rlip cDNA transfected HT22 cells. We found cell survival was decreased in mTau-HT22 cells and RNA-silenced HT22 cells. However, cell survival was increased in Rlip-overexpressed mTau-HT22 cells. A significantly reduced oxygen consumption rate (OCR) was found in mTau-HT22 cells and in RNA-silenced Rlip-HT22 cells, with an even greater reduction in mTau-HT22 + Rlip RNA-silenced HT22 cells. A significantly increased OCR was found in Rlip-overexpressed HT22 cells and in all groups of cells that overexpress Rlip cDNA. Mitochondrial function was defective in mTau-HT22 cells, RNA silenced Rlip in HT22 cells, and was further defective in mTau-HT22 + Rlip RNA-silenced HT22 cells; however, it was rescued in Rlip overexpressed in all groups of HT22 cells. Synaptic and mitophagy proteins were decreased in mTau-HT22 cells, and further reductions were found in RNA-silenced mTau-HT22 cells. However, these were increased in mTau + Rlip-overexpressed HT22 cells. An increased number of mitochondria and decreased mitochondrial length were found in mTau-HT22 cells. These were rescued in Rlip-overexpressed mTau-HT22 cells. These observations strongly suggest that Rlip deficiency causes oxidative stress/mitochondrial dysfunction and Rlip overexpression reverses these defects. Overall, our findings revealed that Rlip is a promising new target for aging, AD, and other tauopathies/neurological diseases.

Coronavirus Disease 2019 Pandemic Effects on Overseas Pakistanis Particularly Residing in China, Saudi Arabia and United Kingdom.

Aim: This study explored the Coronavirus Disease 2019 (COVID-19) pandemic effects on overseas Pakistani's residing in various parts of the world, particularly in China, Saudi Arabia (SA), and the United Kingdom (UK). Methods: This cross-sectional study was completed between November 2020 and April 2021. An online questionnaire was designed and circulated via various social media mediums to overseas Pakistani communities. The obtained data were statistically analyzed through SPSS version 19 for windows. P < 0.05 was considered statistically significant. Results: A total of 542 overseas Pakistani participated in the current study. In total, 157 (29%) were females and 385 (71%) males. There were 174 (32.1%), 142 (26.45%), 117 (21.6%), and 109 (19.85%) participants from the UK, SA, China, and other countries respectively. Some participants, or their family members, 93 (17.2%), got infected with the COVID-19. About one-third, 165 (30.4%), of the respondents were afraid that their company would violate their contracts or lose their jobs or be paid less. The majority, 469 (86.5%), believed that the lockdown is increasing their psychological stress. More than half of the participants, 314 (57.9%), stated that the Pakistani embassy did not facilitate them in their country of stay. About one-third, 194 (35.8%), of the respondents faced visa-related issues. More than one-third of respondents, 221 (40.8%), faced health issues due to lack of physical activities during the lockdown. Males were afraid that their company would violate their contract, lose jobs, or be paid less than females (p < 0.001). Both genders had psychological stress and health issues because of the pandemic. The participants from SA faced more visa-related issues, and they were less satisfied with the efforts of the Pakistani embassy to facilitate them compared to the UK and China participants (p = 0.013). Conclusion: Our data indicate that the COVID-19 pandemic impacted Pakistanis living in the UK, SA, China, and other parts of the world. They had health-related issues, visa problems and dissatisfaction with Pakistani embassy facilitations. Pakistanis living abroad require government assistance to resolve their issues.

COVID-19 Pandemic Awareness, Attitudes, and Practices Among the Pakistani General Public.

Background: Outbreak of COVID-19, in many countries, has imposed a lockdown on their residents. The usefulness of extenuative actions is extremely reliant on society's knowledge, attitudes, and practices (KAP) toward pandemic control. Objective: This study aimed to explore the awareness, attitudes, and practices of the general Pakistani population to COVID-19. Methods: From June 13, 2020, until June 30, 2020, a cross-sectional online KAP survey was conducted among the Pakistani public. For data collection, a validated self-administered questionnaire was used. The survey instrument consisted of six demographic characteristics, 14 items on knowledge, four on attitudes, and six items on practices, modified from a previously published questionnaire on COVID-19. Results: The present study included 2,307 participants, 58.3% males and 41.7% of females. The majority (86.7%) sought information from social media (SM) and television, 95% had good practices, 89.9% had positive attitudes, and two-thirds (67.4%) of the respondents had adequate knowledge. The students and people from younger age groups had more positive attitudes compared with others. Highly educated w with other groups (p < 0.001). In logistic regression analysis, the odds ratio indicated that the private job was negatively associated, and high monthly income was positively associated with adequate knowledge (OR = 0.595). Old age was the predictor of negative attitude, and high school degrees and master's degrees were associated with good practice scores. Conclusion: The Pakistani general population has an overall positive attitude and proactive practices against COVID-19, but their knowledge is inadequate. The most important source of information was SM, followed by television. These are playing a crucial role in educating the Pakistani public.