Antiretroviral Penetration and Drug Transporter Concentrations in the Spleens of Three Preclinical Animal Models and Humans.

Adequate antiretroviral (ARV) concentrations in lymphoid tissues are critical for optimal antiretroviral therapy (ART). While the spleen contains 25% of the body's lymphocytes, there are minimal data on ARV penetration in this organ. This study quantified total and protein-unbound splenic ARV concentrations and determined whether drug transporters, sex, or infection status were modifiers of these concentrations in animal models and humans. Two humanized mice models (hu-HSC-Rag [n = 36; 18 HIV-positive (HIV+) and 18 HIV-negative (HIV-)] and bone marrow-liver-thymus [n = 13; 7 HIV+ and 6 HIV-]) and one nonhuman primate (NHP) model (rhesus macaque [n = 18; 10 SHIV+ and 8 SHIV-]) were dosed to steady state with ARV combinations. HIV+ human spleens (n = 14) from the National NeuroAIDS Tissue Consortium were analyzed postmortem (up to 24 h postdose). ARV concentrations were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS), drug transporter concentrations were measured with LC-MS proteomics, and protein binding in NHP spleens was determined by rapid equilibrium dialysis. Mice generally had the lowest splenic concentrations of the three species. Protein binding in splenic tissue was 6 to 96%, compared to 76 to 99% in blood plasma. NHPs had quantifiable Mrp4, Bcrp, and Ent1 concentrations, and humans had quantifiable ENT1 concentrations. None significantly correlated with tissue ARV concentrations. There was also no observable influence of infection status or sex. With these dosing strategies, NHP splenic penetration most closely resembled that of humans. These data can inform tissue pharmacokinetic scaling to humans to target HIV reservoirs by identifying important species-related differences.

Antiretroviral Penetration across Three Preclinical Animal Models and Humans in Eight Putative HIV Viral Reservoirs.

For HIV cure strategies like "kick and kill" to succeed, antiretroviral (ARV) drugs must reach effective concentrations in putative viral reservoirs. We characterize penetration of six ARVs in three preclinical animal models and humans. We found that standard dosing strategies in preclinical species closely mimicked tissue concentrations in humans for some, but not all, ARVs. These results have implications for interpreting HIV treatment, prevention, or cure interventions between preclinical and clinical models.

The Effect of Blood Loss on Cefazolin Levels in Women Undergoing Cesarean Delivery.

OBJECTIVE: To quantify the effects of operative blood loss during cesarean on tissue and plasma cefazolin concentrations. STUDY DESIGN: This was a prospective observational study of singleton pregnancies undergoing scheduled cesarean between 34 and 40 weeks. Cefazolin administered prior to skin incision. Maternal plasma samples were obtained (Time 1[T1]: immediately, T2: 20 minutes, T3: 40 minutes, and T4: 60 minutes after cefazolin infusion). Subcutaneous adipose tissue sampled before and after fascia. Primary outcome was subcutaneous adipose cefazolin level after fascial closure. Formal quantitative blood loss (QBL) performed. Women with higher QBL, those at/above 75% of QBL in this population, were compared with those with lower QBL (QBL below 75%). Data analyzed using bivariable statistics. RESULTS: Ninety-two women were screened, 32 were eligible, and 20 enrolled. Median QBL was 630 mL (interquartile range [IQR]: 473-818) and 1,160 mL (IQR: 1,000-1,560) in the low and high QBL groups, respectively. Demographics and operative characteristics were similar. Median adipose cefazolin level after fascial closure did not differ between the groups (3.5 vs. 3.9 µg/g, p = 0.75). No differences in maternal plasma cefazolin concentrations between the groups at any time point or in pharmacokinetic parameters were seen. CONCLUSION: Intraoperative maternal plasma concentrations and adipose levels of cefazolin are similar between women with high and low blood loss at the time of cesarean delivery.

Development and validation of an LC-MS/MS assay for the quantification of dolutegravir extracted from human hair.

Measurement of drug concentrations in hair provides a non-invasive approach to assess drug adherence. Here, we report on the development and validation of a method for the quantification of the antiretroviral dolutegravir (DTG) extracted from human hair. DTG is extracted from hair samples by sonication and incubation in 50:50 methanol:acetonitrile with 2% formic acid overnight at 40 °C. Following extraction, samples are analyzed by reverse-phase chromatography on a Waters Atlantis T3 (50 × 2.1 mm, 3-µm particle size) column with subsequent detection by electrospray ionization in positive ion mode on an AB Sciex API-5000 triple quadrupole mass spectrometer. The stable, isotopically labeled 13C,d5-DTG is used as an internal standard in the assay. The calibration range is 5-10,000 pg DTG/mL of extraction solvent with the ability to extract between 1 and 10 mg of hair/mL of extraction solvent. The assay was linear, accurate (inter-assay %bias within ± 6.5%), and precise (inter-assay %CV ≤ 10.3%). The assay was successfully used to analyze clinical samples from subjects on DTG regimens. Analysis of clinical samples suggested the potential presence of a degradation product, which was subsequently confirmed to occur with exposure to sunlight. The degradation of DTG could complicate absolute interpretation of clinical results, but the presence of this degradation product is easily evaluated with this assay to aid in data interpretation.

Advanced brain ageing in adult psychopathology: A systematic review and meta-analysis of structural MRI studies.

Evidence suggests that psychopathology is associated with an advanced brain ageing process, typically mapped using machine learning models that predict an individual's age based on structural neuroimaging data. The brain predicted age difference (brain-PAD) captures the deviation of brain age from chronological age. Substantial heterogeneity between studies has introduced uncertainty regarding the magnitude of the brain-PAD in adult psychopathology. The present meta-analysis aimed to quantify structural MRI-based brain-PAD in adult psychotic and mood disorders, while addressing possible sources of heterogeneity related to diagnosis subtypes, segmentation method, age and sex. Clinical factors influencing brain ageing in axis 1 psychiatric disorders were systematically reviewed. Thirty-three studies were included for review. A random-effects meta-analysis revealed a brain-PAD of +3.12 (standard error = 0.49) years in psychotic disorders (n = 16 studies), +2.04 (0.10) years in bipolar disorder (n = 5), and +0.90 (0.20) years in major depression (n = 7). An exploratory meta-analysis found a brain-PAD of +1.57 (0.67) in first episode psychosis (n = 4), which was smaller than that observed in psychosis and schizophrenia (n = 10, +3.87 (0.61)). Patient mean age significantly explained heterogeneity in effect size estimates in psychotic disorders, but not mood disorders. The systematic review determined that clinical factors, such as higher symptom severity, may be associated with a larger brain-PAD in psychopathology. In conclusion, larger structural MRI-based brain-PAD was confirmed in adult psychopathology. Preliminary evidence was obtained that brain ageing is greater in those with prolonged duration of psychotic disorders. Accentuated brain ageing may underlie the cognitive difficulties experienced by some patients, and may be progressive in nature.

Genital Mucosal Drug Concentrations and anti-HIV Activity in Tenofovir-Based PrEP Products: Intravaginal Ring vs. Oral Administration.

OBJECTIVE: To describe and compare systemic and local pharmacokinetics (PK) and cervicovaginal (CV) pharmacodynamics (PD) of oral tenofovir disoproxil fumarate (TDF) in combination with emtricitabine (FTC) with tenofovir (TFV) intravaginal ring (IVR). DESIGN: Phase I, randomized, parallel-group study. Women (n = 22) used TDF/FTC oral tablets daily or TFV IVR continuously and were assessed at baseline and 14 days. METHODS: TFV and FTC concentrations were measured in plasma, CV fluid (CVF), and CV tissue. TFV-diphosphate and FTC-triphosphate were assessed in CV tissue. In vitro PD antiviral activities of TFV and FTC (using in vivo concentration ranges) were modeled in the CVF and by infecting CV tissue explants ex vivo with HIV-1BaL. RESULTS: Adverse events (AEs) were more common with oral TDF/FTC use (P < 0.01). The median CVF TFV concentrations were 106 ng/mL after use of TFV IVR vs. 102 ng/mL for TDF/FTC. The median TFV and TFV-diphosphate concentrations in CV tissue were >100-fold higher among IVR users. The median CVF FTC concentrations were 103 ng/mL. FTC and FTC-triphosphate were detected in all CV tissues from TDF/FTC users. HIV inhibitory activity of CVF increased significantly with treatment in both cohorts (P < 0.01) but was higher in TFV IVR users (P < 0.01). In vitro inhibition of tissue infection with ex vivo administration of TFV and FTC was dose dependent, with maximal efficacy achieved with 10 µg/mL TFV, 1 µg/mL FTC, and 0.1 µg/mL of TFV and FTC combined. CONCLUSIONS: Both products were safe and increased mucosal HIV inhibitory activity. In addition to systemic protection, oral TDF/FTC displays a PK/PD profile compatible with CV mucosal antiviral activity. TFV IVR resulted in fewer AEs, lower TFV plasma concentrations, higher CVF and tissue TFV and TFV-DP concentrations, and greater anti-HIV activity in CVF.

Perceived barriers to provision of medication therapy management services (MTMS) and the likelihood of a pharmacist to work in a pharmacy that provides MTMS.

BACKGROUND: Recently, medication therapy management services (MTMS) has gained significant attention as an important type of pharmaceutical care designed to improve patient outcomes with more appropriate medication usage and monitoring. Although the provision of MTMS is increasing in pharmacies across the nation, and pharmacists are well equipped to administer MTMS, many community pharmacists are not currently providing these services. OBJECTIVE: To determine barriers to provision of MTMS perceived by pharmacists and factors associated with the likelihood of working in a pharmacy that provides MTMS. METHODS: Surveys were mailed to 906 community pharmacists licensed in West Virginia using a stratified random sample. The instrument was constructed and finalized following a review by experts and pilot tested in a convenience sample of pharmacists. Principal components analysis was performed to determine the factors that describe perceived barriers to provision of MTMS. Discriminant analysis using factor scores and other demographic and practice variables was performed to predict respondents' likelihood to work in a pharmacy that provides MTMS. RESULTS: A 3-factor model was extracted from the responses, which explained 53.3% of the total variance. The factors included perceived ability to respond to patient interest, pharmacy-related factors, and enabling factors. The discriminant function correctly classified 76.2% of cases and included comfort level with provision of services, perceived value of services to patients, perceived ability to respond to patient interest, and whether they currently offer MTMS. These variables were all positively correlated with pharmacists' likelihood of working in a pharmacy that provides MTMS. CONCLUSIONS: Comfort level and ability are important factors that influence pharmacists' likelihood of working in a pharmacy that provides MTMS. These findings highlight the importance of advanced practice experiences, certificate programs, and residencies to build pharmacists' confidence, and the role of targeted recruitment to attract pharmacists to community pharmacies that provide MTMS.

Medication therapy management services in West Virginia: pharmacists' perceptions of educational and training needs.

BACKGROUND: The Medicare Modernization Act of 2003 recognizes the challenges associated with drug therapy in elderly patients with multiple chronic diseases, and requires the development of medication therapy management services (MTMS) for such beneficiaries. OBJECTIVE: To assess pharmacists' perception of educational and training needs necessary to implement MTMS in community pharmacies in West Virginia, USA. METHODS: Self-administered mail surveys with an explanatory cover letter were mailed to the designated pharmacist-in-charge (PIC) of each licensed community pharmacy (506) in West Virginia. Main outcome measures included pharmacists' comfort level, perceptions of value to patients, barriers to provision of services, and pharmacists' interest in receiving education and training related to MTMS. RESULTS: Of the 503 surveys that were deliverable, 203 (40.4%) usable responses were received. Fifty-five (27.1%) PICs reported that MTMS are currently being provided in their pharmacy. Respondents were likely to use services that aid in the development of MTMS and disease-state management, felt relatively comfortable in providing MTMS, and had a favorable view of the value of services to patients, but reported that lack of time tended to be a barrier. CONCLUSION: PICs in West Virginia are interested in and open to their pharmacists receiving education and training for implementation of MTMS.

Population Modeling Highlights Drug Disposition Differences Between Tenofovir Alafenamide and Tenofovir Disoproxil Fumarate in the Blood and Semen.

Understanding antiretroviral disposition in the male genital tract, a distinct viral compartment, can provide insight for the eradication of HIV. Population pharmacokinetic modeling was conducted to investigate the disposition of tenofovir disoproxil fumarate (TDF), tenofovir alafenamide (TAF), and emtricitabine and their metabolites in blood and semen. Blood plasma and seminal plasma (SP) concentrations of tenofovir and emtricitabine were measured, as were tenofovir-diphosphate and emtricitabine-triphosphate concentrations in peripheral blood mononuclear cells (PBMCs) and seminal mononuclear cells. Sequential compartmental modeling described drug disposition in blood and semen. Our modeling suggests slower elimination of apparent tenofovir-diphosphate PBMC and faster elimination of tenofovir SP after administration of TAF compared with TDF, likely reflecting flip-flop kinetics. Additionally, TAF metabolism to tenofovir appeared slower in semen compared with blood; however, SP elimination of TAF-derived tenofovir appeared faster than its blood plasma elimination. These findings provide valuable insight for further mechanistic study of cellular entry and drug metabolism in the male genital tract.

Differential extracellular, but similar intracellular, disposition of two tenofovir formulations in the male genital tract.

BACKGROUND: The male genital tract (MGT) is a viral sanctuary and likely HIV reservoir; understanding MGT pharmacokinetics (PK) of antiretrovirals (ARVs) used for curative strategies is critical to eradication and cure. Tenofovir alafenamide (TAF) is a tenofovir (TFV) formulation designed to maximize efficacy/minimize toxicity with unknown MGT PK. METHODS: HIV-positive and HIV-negative men receiving TFV-based regimens provided six paired blood plasma (BP) and semen samples. Extracellular (TFV, TAF, emtricitabine [FTC]) drug concentrations in BP and seminal plasma (SP), and intracellular metabolite (IM) and endogenous nucleotide (EN) concentrations were measured in peripheral blood mononuclear cells (PBMCs) and seminal mononuclear cells (SMCs). Exposure ratios for SP:BP, SMC:PBMC and IM:EN were calculated from PK parameters generated by noncompartmental analysis. HIV viral load was measured in BP and SP. RESULTS: Sixteen HIV-positive (n=8, TDF/FTC; n=8, TAF/FTC) and eight HIV-negative (TDF/FTC) men provided samples. Median TFV SP:BP ratios differed between TDF and TAF (1.5 versus 7.4), due to lower TFV BP concentrations with TAF coupled with TFV SP concentrations similar to TDF. FTC SP: BP ratios were approximately 3. SMC concentrations of IMs and ENs were a fraction of PBMC concentrations (1-22%), though IM:EN ratios exceed a suggested protective threshold. CONCLUSIONS: TAF SP PK was unexpected. IM SMC concentrations were low relative to PBMC, as were EN concentrations, suggesting differences in cell phenotype and lineage in the MGT; these differences in phenotype and pharmacology may have an impact on selecting and dosing ARVs used in cure strategies.

Predicting Efavirenz Concentrations in the Brain Tissue of HIV-Infected Individuals and Exploring their Relationship to Neurocognitive Impairment.

Sparse data exist on the penetration of antiretrovirals into brain tissue. In this work, we present a framework to use efavirenz (EFV) pharmacokinetic (PK) data in plasma, cerebrospinal fluid (CSF), and brain tissue of eight rhesus macaques to predict brain tissue concentrations in HIV-infected individuals. We then perform exposure-response analysis with the model-predicted EFV area under the concentration-time curve (AUC) and neurocognitive scores collected from a group of 24 HIV-infected participants. Adult rhesus macaques were dosed daily with 200 mg EFV (as part of a four-drug regimen) for 10 days. Plasma was collected at 8 time points over 10 days and at necropsy, whereas CSF and brain tissue were collected at necropsy. In the clinical study, data were obtained from one paired plasma and CSF sample of participants prescribed EFV, and neuropsychological test evaluations were administered across 15 domains. PK modeling was performed using ADAPT version 5.0 Biomedical Simulation Resource, Los Angeles, CA) with the iterative two-stage estimation method. An eight-compartment model best described EFV distribution across the plasma, CSF, and brain tissue of rhesus macaques and humans. Model-predicted median brain tissue concentrations in humans were 31 and 8,000 ng/mL, respectively. Model-predicted brain tissue AUC was highly correlated with plasma AUC (γ = 0.99, P < 0.001) but not CSF AUC (γ = 0.34, P = 0.1) and did not show any relationship with neurocognitive scores (γ < 0.05, P > 0.05). This analysis provides an approach to estimate PK the brain tissue in order to perform PK/pharmacodynamic analyses at the target site.

The efficacy of telephone counseling for health promotion in people with multiple sclerosis: a randomized controlled trial.

OBJECTIVE: To determine if motivational interviewing-based telephone counseling increases health promotion activities and improves other health outcomes in people with multiple sclerosis (MS). DESIGN: Randomized controlled trial with wait-list controls and single-blinded outcome assessments conducted at baseline and at 12 weeks. SETTING: MS research and training center in the Pacific Northwest. PARTICIPANTS: Community-residing persons (N=130) with physician confirmed MS aged 18 or older who were able to walk unassisted at least 90 m (300 ft). INTERVENTION: A single in-person motivational interview followed by 5 scheduled telephone counseling sessions to facilitate improvement in 1 of 6 health promotion areas: exercise, fatigue management, communication and/or social support, anxiety and/or stress management, and reducing alcohol or other drug use. MAIN OUTCOME MEASURES: Health Promotion Lifestyle Profile II plus fatigue impact, subjective health, and objective measures of strength, fitness, and cognition. Intent-to-treat analyses of change scores were analyzed using nonparametric tests. RESULTS: Seventy persons were randomized to treatment and 60 to the control condition. The treatment group reported significantly greater improvement in health promotion activities, including physical activity, spiritual growth, and stress management as well as in fatigue impact and mental health compared with controls. In addition, the exerciser subgroup showed greater improvement than controls in self-selected walking speed. CONCLUSIONS: A less intensive, more accessible approach to health promotion based on telephone counseling and motivational interviewing shows promise and merits further study.

Associations among cardiometabolic risk factor clustering, weight status, and cardiovascular disease in an Appalachian population.

It has been suggested that within the traditional body mass index (BMI) categories there is a heterogeneous pattern of cardiometabolic risk factor clustering. The objective of this research was to determine the associations among obesity, cardiometabolic abnormalities, and cardiovascular disease (CVD) in a large population-based study of Appalachian adults. The study comprised a cross-sectional survey of Appalachian adults residing in 6 communities in Ohio and West Virginia, who were aged 18 years and older (n=14,783, 50.9% women). The authors categorized BMI into normal weight (<25kg/m(2) ), overweight (25-29.9kg/m(2) ), and obese (≥30kg/m(2) ). Cardiometabolic abnormalities were defined as the presence of hypertension, elevated triglycerides (≥150mg/dL), decreased high-density lipoprotein cholesterol (<40mg/dL [men], <50mg/dL [women]), elevated fasting glucose (≥100mg/dL)/diabetes, insulin resistance (homeostasis model assessment >5.13), or elevated C-reactive protein (>3mg/L). They found that 25.6% of normal-weight adults displayed clustering of ≥2 cardiometabolic abnormalities; in contrast, 36.8% of overweight/obese adults displayed no clustering. Compared with normal-weight persons without clustering of cardiometabolic abnormalities (referent), the odds ratio of CVD was 1.06 (95% confidence interval [CI], 0.84-1.34) among overweight/obese individuals without cardiometabolic clustering, 2.21 (95% CI, 1.74-2.81) among normal-weight individuals with cardiometabolic clustering, and 2.45 (95% CI, 2.02-2.97) among overweight/obese individuals with cardiometabolic clustering. These results suggest that within the traditional BMI categories, there may be heterogeneity of CVD risk depending on whether there is underlying clustering of cardiometabolic abnormalities.

Attributions for relatives' behavior and perceived criticism: studies with community participants and patients with anxiety disorders.

The relationship between perceived criticism from one's relative and attributions about that relative's behavior was examined in two studies. In Study 1, 50 community couples volunteered to participate in a study of marital interaction. Participants rated their interaction-specific perceived criticism after a 10-min problem-solving interaction and their attributions for their spouses' behavior during a review of the videotaped interaction. In Study 2, 70 outpatients with obsessive-compulsive disorder (n=41) or panic disorder with agoraphobia (n=29) completed a measure of global perceived criticism in their relationship with their spouse or other family member and on another occasion participated in a 10-min problem-solving interaction with that relative. Using interaction transcripts, coders extracted and coded attributions from patients' speech and, using the videotapes themselves, rated relatives' observable criticism. In both studies higher scores on negative attributions were related to higher perceived criticism ratings. In Study 2, negative attributions contributed to the prediction of perceived criticism above and beyond the contribution of observed criticism. These findings suggest that targeting attributions about perceived criticism may be fruitful in reducing the negative impact of perceived criticism on treatment outcome for a variety of psychiatric disorders.

Construct validity of the Perceived Criticism Measure.

The construct validity of the Perceived Criticism Measure (PCM) was examined in 2 studies. In Study 1, 50 community couples participated in problem-solving interactions after which they rated interaction-specific perceived criticism and their criticism of their spouses. In addition, they provided ratings of perceived criticism for their relationship overall and completed measures of psychopathology and marital satisfaction. For both husbands and wives, convergent validity was demonstrated by moderate-to-large correlations between the PCM and spouses' own ratings of their criticism for both general and interaction-specific perceived criticism. In Study 2, 37 patients with obsessive-compulsive disorder and their spouses participated in problem-solving interactions and provided ratings of marital satisfaction and general perceived criticism. Five untrained coders rated the interactions according to their own definitions of the relatives' destructive criticism of the patient. Their aggregated ratings proved strongly related to patients' PCM scores. Higher PCM scores were related to lower marital satisfaction in both Studies 1 and 2. The results of these studies are supportive of the convergent validity of the Perceived Criticism Measure. Evidence of discriminant validity was mixed.