Monochrome Multiplex Quantitative PCR Telomere Length Measurement.

Telomeres are ribonucleoprotein structures at the end of all eukaryotic chromosomes that protect DNA from damage and preserve chromosome stability. Telomere length (TL) has been associated with various exposures, biological processes, and health outcomes. This article describes the monochrome multiplex quantitative polymerase chain reaction (MMqPCR) assay protocol routinely conducted in our laboratory for measuring relative mean TL from human DNA. There are several different PCR-based TL measurement methods, but the specific protocol for the MMqPCR method presented in this publication is repeatable, efficient, cost-effective, and suitable for population-based studies. This detailed protocol outlines all information necessary for investigators to establish this assay in their laboratory. In addition, this protocol provides specific steps to increase the reproducibility of TL measurement by this assay, defined by the intraclass correlation coefficient (ICC) across repeated measurements of the same sample. The ICC is a critical factor in evaluating expected power for a specific study population; as such, reporting cohort-specific ICCs for any TL assay is a necessary step to enhance the overall rigor of population-based studies of TL. Example results utilizing DNA samples extracted from peripheral blood mononuclear cells demonstrate the feasibility of generating highly repeatable TL data using this MMqPCR protocol.

A review and meta-analysis: Cross-tissue telomere length correlations in healthy humans.

BACKGROUND AND AIM: Tissue source has been shown to exert a significant effect on the magnitude of associations between telomere length and various health outcomes and exposures. The purpose of the present qualitative review and meta-analysis is to describe and investigate the impact of study design and methodological features on the correlation between telomere lengths measured in different tissues from the same healthy individual. METHODS: This meta-analysis included studies published from 1988 to 2022. Databases searched included PubMed, Embase, and Web of Science and studies were identified using the keywords "telomere length" and "tissues" or "tissue." A total of 220 articles of 7856 initially identified studies met inclusion criteria for qualitative review, of which 55 met inclusion criteria for meta-analysis in R RESULTS: Studies meeting inclusion criteria for meta-analysis tended to have enhanced demographic and methodological reporting relative to studies only included in the qualitative review. A total of 463 pairwise correlations reported for 4324 unique individuals and 102 distinct tissues were extracted from the 55 studies and subject to meta-analysis, resulting in a significant effect size z = 0.66 (p < 0.0001) and meta-correlation coefficient of r = 0.58. Meta-correlations were significantly moderated by sample size and telomere length measurement methodology, with studies of smaller size and those using hybridization-based analyses exhibiting the largest meta-correlation. Tissue source also significantly moderated the meta-correlation, wherein correlations between samples of a different lineage (e.g., blood vs. non-blood) or collection method (e.g., peripheral vs. surgical) were lower than correlations between samples of the same lineage or collection method. CONCLUSION: These results suggest that telomere lengths measured within individuals are generally correlated, but future research should be intentional in selecting a tissue for telomere length measurement that is most biologically relevant to the exposure or outcome investigated and balance this with the feasibility of obtaining the sample in sufficient numbers of individuals.

Wisconsin Healthcare Utilization Cost Among American Indians/Alaska Natives with and without Alzheimer's Disease and Related Dementias.

Individuals with Alzheimer's disease and related dementias (ADRD) accrue higher healthcare utilization costs than peers without ADRD, but incremental costs of ADRD among American Indians/Alaska Natives (AI/AN) is unknown. State-wide paid electronic health record data were retrospectively analyzed using percentile-based bootstrapped 95% confidence intervals of the weighted mean difference of total 5-year billed costs to compare total accrued for non-Tribal and Indian Health Service utilization costs among Medicaid and state program eligible AI/AN, ≥40 years, based on the presence/absence of ADRD (matching by demographic and medical factors). AI/AN individuals with ADRD accrued double the costs compared to those without ADRD, costing an additional $880.45 million to $1.91 billion/year.

Method comparison studies of telomere length measurement using qPCR approaches: A critical appraisal of the literature.

Use of telomere length (TL) as a biomarker for various environmental exposures and diseases has increased in recent years. Various methods have been developed to measure telomere length. Polymerase chain reaction (PCR)-based methods remain wide-spread for population-based studies due to the high-throughput capability. While several studies have evaluated the repeatability and reproducibility of different TL measurement methods, the results have been variable. We conducted a literature review of TL measurement cross-method comparison studies that included a PCR-based method published between January 1, 2002 and May 25, 2020. A total of 25 articles were found that matched the inclusion criteria. Papers were reviewed for quality of methodologic reporting of sample and DNA quality, PCR assay characteristics, sample blinding, and analytic approaches to determine final TL. Overall, methodologic reporting was low as assessed by two different reporting guidelines for qPCR-based TL measurement. There was a wide range in the reported correlation between methods (as assessed by Pearson's r) and few studies utilized the recommended intra-class correlation coefficient (ICC) for assessment of assay repeatability and methodologic comparisons. The sample size for nearly all studies was less than 100, raising concerns about statistical power. Overall, this review found that the current literature on the relation between TL measurement methods is lacking in validity and scientific rigor. In light of these findings, we present reporting guidelines for PCR-based TL measurement methods and results of analyses of the effect of assay repeatability (ICC) on statistical power of cross-sectional and longitudinal studies. Additional cross-laboratory studies with rigorous methodologic and statistical reporting, adequate sample size, and blinding are essential to accurately determine assay repeatability and replicability as well as the relation between TL measurement methods.

Methylphenidate and alcohol effects on flash-evoked potentials, body temperature, and behavior in Long-Evans rats.

Methylphenidate (MPD) is a psychostimulant used to treat attention deficit hyperactivity disorder (ADHD). Most adult ADHD patients use ethanol in combination with MPD. This research examined the effects of MPD and ethanol on flash-evoked potentials (FEPs; cortical responses frequently used to assess neural activity and sensory processing) recorded from the visual cortex (VC) and superior colliculus (SC; a structure involved in attention and orientation) of chronically implanted male Long-Evans rats, and on body temperature and open field behavior. For one group of rats, either saline or ethanol (2.0 g/kg) was given 5 min prior to either saline or MPD (2.9 mg/kg). FEPs were recorded 10 and 20 min later. In the VC, ethanol decreased amplitudes of several components, but increased P2. MPD increased N3, but decreased P3 and P4. Ethanol increased the latency of several components. In the SC, ethanol decreased all three components, while MPD increased P3. Ethanol increased latency of all components. During FEP testing, ethanol decreased body movement while MPD increased movement. In the open field, line crossings were increased but rearings were decreased by ethanol. Both ethanol and MPD produced hypothermia. A second group of rats was given MPD at 11.6 mg/kg. Ethanol decreased several VC amplitudes, but increased P2. MPD increased N3 amplitude but decreased amplitude for other components. MPD also counteracted the effect of ethanol on the amplitude of P2 and N3. Both ethanol and MPD increased the latency of several components. In the SC, ethanol decreased all component amplitudes, while MPD increased P3 but decreased N4. Ethanol increased all component latencies, while MPD increased latency for two components. During FEP testing, ethanol decreased body movement while MPD increased movement. In the open field, line crossings were increased by ethanol and MPD. Rearings were eliminated by ethanol in the open field but increased by MPD, and MPD counteracted the effect of ethanol on rearings. Both ethanol and MPD produced hypothermia. Some of these results might help explain why users take MPD and ethanol in combination in order to enable consuming larger amounts of alcohol.

Reactions of CF3CH2I+O(3P): competing mechanisms of HF elimination.

Ab initio density functional and molecular orbital calculations provide singlet and triplet electronic potential energy surfaces for the reactions of CF3CH2I+O(3P) leading to OI and HF eliminations, reactions which have been the subject of recent experimental studies. A barrier to OI formation occurs on the triplet potential energy surface; there is no reverse barrier to OI formation on the singlet pathway. Findings suggest that two competing pathways may form HF. One is an addition-insertion-elimination process involving insertion of O into the C-I bond. The alternate path involves OI elimination, addition of an O atom to CF3CH2, and subsequent HF elimination. The computed reactant pathways and energetics are discussed in relation to recent experiments.