A new, sensitive and fast method for the simultaneous determination of pyrazinamide, isoniazid, streptomycin, ethambutol, and rifampicin in human plasma was developed and validated. The method required only 100 μL of plasma and one step for sample preparation by protein precipitation. The drugs were separated by using a hydrophilic interaction liquid chromatography (HILIC) column. The mobile phase was methanol and water (0.1% formic acid and 5 mM ammonium acetate, pH 3.0 ± 0.1) in a ratio of 65:35 (v/v), which was eluted at an isocratic flow rate of 0.5 mL/min. Tandem mass spectrometry was performed with a triple-quadrupole tandem mass spectrometer. By use of the HILIC column, the detection was free of ion-pair reagents in the mobile phase, with no significant matrix effects. The total run time was less than 2 min for each sample. The method was validated by evaluating its selectivity, sensitivity, linearity, accuracy, and precision according to US Food and Drug Administration guidelines. The lower limit of quantification was 4.0 ng/mL for pyrazinamide, isoniazid, and rifampicin, 0.5 ng/mL for ethambutol, and 10.0 ng/mL for streptomycin. The intraday precision and interday precision were less than 9%, with the accuracy ranging between -9.3 and 7.3%. The method was successfully applied to therapeutic drug monitoring of 33 patients with tuberculosis after administration of standard antituberculosis drugs. The method has been proved to meet the high-throughput requirements in therapeutic drug monitoring.

Escherichia coli O157:H7 is well known for many foodborne outbreaks that lead to fatal infections in human being worldwide. The objective of this study was to develop a rapid and sensitive method for detection of EHEC O157:H7 from ground beef using a method that combined immunomagnetic separation (IMS) with loop-mediated isothermal amplifi cation (LAMP). The EHEC O157:H7 cells were separated with Dynabeads coated with anti-EHEC O157:H7 after a short enrichment for 4 h. Then, EHEC O157:H7 was identifi ed by LAMP assay for amplifying and detecting the rfbE gene, which is highly conserved in all EHEC O157:H7 strains and exhibits strain-specifi c gene expression. The LAMP method results analyzed with real time turbidity measurements showed a high specifi city and sensitivity, with a positive detection rate of amplifi cation of EHEC O157:H7 DNA diluted to a minimum equivalent concentration of 1.8 × 10 1 CFU/mL, which was 10 times more sensitive than the conventional PCR assay. The IMS followed with LAMP could capture and detect a bacterial concentration as low as 3×10 1 CFU/mL from the meat samples, which was close to the sensitivity of LAMP assay with pure culture. IMS combined with realistic LAMP method is a simple, rapid, highly specifi c gene amplifi cation technology that is suitable for implementing as a screening assay in basic laboratory and fi eld test for detecting food contamination. Unauthenticated Download Date | 5/11/18 5:54 AM

OBJECTIVE To examine the association between microvascular disease (MVD) and risk of heart failure (HF) among individuals with type 1 diabetes mellitus (T1DM) or type 2 diabetes mellitus (T2DM). RESEARCH DESIGN AND METHODS We included 1,713 and 28,624 participants with T1DM and T2DM, respectively, from the UK Biobank who were free of HF during enrollment. MVD burden reflected by the presence of retinopathy, peripheral neuropathy, and chronic kidney disease (CKD) at baseline was prospectively evaluated for the association with incidence of HF. Hazard ratios (HRs) and 95% CIs of HF were estimated by Cox regression models adjusted for multiple traditional risk factors. RESULTS There were 145 and 2,515 incident cases of HF recorded among participants with T1DM and T2DM, respectively, during a median follow-up of 11.5 years. The association between the number of MVD and HF was stronger among participants with T1DM than among those with T2DM (P for interaction <0.001). Compared with participants with no MVD, those with all three MVD had an adjusted HR (95% CI) of 11.37 (5.62, 22.99) in T1DM and 3.66 (2.74, 4.88) in T2DM. In T1DM, HRs (CIs) were 2.69 (1.75, 4.14) for retinopathy, 2.11 (1.38, 3.23) for peripheral neuropathy, and 2.21 (1.53, 3.18) for CKD. The corresponding estimates in T2DM were 1.24 (1.13, 1.36), 1.63 (1.36, 1.96), and 1.73 (1.59, 1.89), respectively. CONCLUSIONS While a heavier burden of MVD was associated with excess risk of HF both in T1DM and T2DM, the association was evidently more pronounced in T1DM.

Importance:The American Heart Association (AHA)’s Life’s Essential 8 (LE8) score, a recently updated metric for promoting cardiovascular health (CVH), has addressed the limitations of the original metrics (Life’s Simple 7) and is able to quantify CVH. Objective:To evaluate the associations of LE8 score with the risk of incident dementia and its subtypes, cognition, and neuroimaging outcomes and to determine whether these associations differ among apolipoprotein E (APOE)-ε4 genotypes. Design, Setting, and Participants:UK Biobank participants without prior cardiovascular disease nor dementia at baseline (2006–2010) were enrolled in this prospective cohort study. Data analysis was conducted from December 20, 2022, to February 15, 2023. Exposures:A modified version of the LE8 score was created (range: 0–100) and categorized into poor (0–49), intermediate (50–79), and optimal (80–100) CVH. Main Outcomes and Measures:The outcomes included incident dementia (all-cause, vascular [VaD], and Alzheimer’s disease ascertained through hospital inpatient and death records), cognitive test scores (fluid intelligence and numeric memory), and neuroimaging markers (total brain volume [BV], white matter hyperintensity [WMH], and hippocampal volume). Adjusted Cox proportional hazard and multivariable linear regression models were used. Results: A subsample of 316,669 participants (mean [SD] age, 56.3 [8.1] years) were included. Higher LE8 scores were associated with reduced risk of all-cause dementia and VaD, the adjusted hazard ratios (HRs) in the optimal CVH versus the poor CVH group were 0.56 (95% confidence interval [CI], 0.48–0.64) and 0.29 (95% CI, 0.22–0.38), respectively. A 10-point increment in LE8 was associated with higher fluid intelligence (β, 0.088; 95% CI, 0.073–0.102) and numeric memory (β, 0.054; 95% CI, 0.043–0.065), and was also associated with lower WMH volume (β, −0.673; 95% CI, from −0.751 to −0.596) and larger BV (β, 77.93; 95% CI, 62.03–93.84) and hippocampal volume (β, 0.197; 95% CI, 0.106–0.288). These associations were more evident in APOE-ε4 noncarriers. Conclusions and Relevance: Individuals with a higher LE8 score experienced less dementia events (driven especially by incident VaD) and were associated with better neurocognitive brain health profiles. CVH optimization may be beneficial to the maintenance of brain health.

Background Higher fluctuation of cholesterol has been shown associated with cognitive decline. But the association between unidirectional variation of cholesterol and cognition remains unclear. In the present study, we examined the association between variation of cholesterol from normal to high level or from high to normal level and cognition. Methods A total of 4,915 participants (mean age, 57.7 years; 49.2% female) with normal cognition in baseline (2011) from the China Health and Retirement Longitudinal Study (CHARLS) were included. Plasma total cholesterol (TC), non-high-density lipoprotein cholesterol (NHDL-C), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were divided into two clusters according to cutoff in 2011 and 2015, respectively. Participants were divided into four groups: normal-normal, normal-high, high-normal and high-high. Cognitive functions were assessed by episodic memory and mental intactness. The binary logistic regression was used to analyze the association of cholesterol variation with cognitive decline. Results Participants with increased cholesterol were more likely had lower incidence of cognitive decline. The fluctuations of cholesterol were more likely occurred in females. The odds ratio (OR) and 95% confidence interval (CI) of global cognitive decline in normal-high compared with normal-normal group of TC was 0.61 (0.36–1.03), and significant only in females [OR and 95% CI: 0.50 (0.26–0.97)]; the risk of memory function decline in high-normal group compared with high-high group was increased both in males [OR and 95% CI: 1.98 (1.02–4.05)] and females [OR and 95% CI: 1.61 (1.01–2.74)]. The similarly results were shown in NHDL-C groups [OR and 95% CI of global cognitive decline in normal-high group: 0.50 (0.26–0.95) for overall; 0.38 (0.17–0.85) for females]; but the risk of memory function decline in high-normal group only increased in females [OR and 95% CI: 1.64 (1.02–2.25)]. The risk of mental intactness decline in males significantly decreased in high-normal group of LDL-C compared with high-high group [OR and 95% CI: 0.30 (0.13–0.67)] and increased in high-high compared with normal-normal group [OR and 95% CI: 2.27 (1.18–2.98)]. Conclusions Longitudinal decreasing of TC or NHDL-C is detrimental to cognitive function, especially in females. But LDL-C is still a risk factor of cognitive decline in males.