1ß-Hydroxydeoxycholic Acid as an Endogenous Biomarker in Human Plasma for Assessment of CYP3A Clinical Drug-Drug Interaction Potential.

4ß-Hydroxycholesterol (4ß-HC) in plasma has been used as a biomarker to assess CYP3A drug-drug interaction (DDI) potential during drug development. However, due to the long half-life and narrow dynamic range of 4ß-HC, its use has been limited to the identification of CYP3A inducers, but not CYP3A inhibitors. The formation of 1ß-hydroxydeoxycholic acid (1ß-OH DCA) from deoxycholic acid (DCA) is mediated by CYP3A, thus 1ß-OH DCA can potentially serve as an alternative to 4ß-HC for assessment of CYP3A DDI potential. To study this feasibility, we developed a sensitive LC-MS/MS method for the simultaneous quantitation of 1ß-OH DCA and its glycine and taurine conjugates in human plasma with the LLOQ of 50 pg/mL, which enabled the quantitation of basal levels and further reduction. The method was applied to a DDI study to assess how 1ß-OH DCA and its glycine and taurine conjugates would respond to CYP3A induction or inhibition. Rifampin induction resulted in an increase of 1ß-OH DCA and its conjugates in plasma, with 6.8-, 7.8-, 8.3-, 10.3-fold increases of AUCLST, AUC24h, Cmax and mean concentrations for total 1ß-OH DCA (total of all three forms), respectively. Importantly, inhibition with itraconazole resulted in notable reduction of these biomarkers, with 84%, 85%, 82%, 81% reductions of AUCLST, AUC24h, Cmax and mean concentrations for total 1ß-OH DCA, respectively. This preliminary data demonstrates for the first time that total 1ß-OH DCA in plasma has the potential to serve as a biomarker for CYP3A DDI assessment in early clinical development and may provide key advantages over 4ß-HC. Significance Statement We have reported the use of total 1ß-Hydroxydeoxycholic Acid (1ß-OH DCA) (sum of 1ß-OH DCA and its glycine and taurine conjugates) plasma concentration as a biomarker for CYP3A activity. Itraconazole inhibition led to an 81-85% decrease of total 1ß-OH DCA plasma exposures, while rifampin induction led to a 6.8-10.3 fold increase of total 1ß-OH DCA plasma exposures. Using 1ß-OH DCA exposures in plasma also provides benefit of allowing PK and biomarker assessment using the same matrix, thus simplify collection procedures.

The "SuperAgers" construct in clinical practice: neuropsychological assessment of illiterate and educated elderly.

INTRODUCTION: The demographic transition is a global event intensified during the last decades that represents population aging. Thus, the studies directed to the elderly 80 years of age or more with preserved cognitive functions (named SuperAgers) emerges as a possible path to full comprehension of the health of those aging with acceptable levels of functionality and independency. OBJECTIVE: To evaluate the cognitive performance of the elderly over 80 years old, associating the results to their educational level. METHOD: We evaluated 144 healthy elders with 80 years or more through the following cognitive tests Mini-Mental State Examination (MMSE), Cambridge Cognitive Examination (CAMCOG), Clock Drawing Test (CDT), and Verbal Fluency Test (VF) and compared the tests' scores with their educational level segmented in years of formal education, being the groups ILLITR (<1 year of schooling), 1TO4 (from 1 to 4 years of schooling), and 5MORE (>5 years of schooling). RESULTS: There was positive influence of educational level on the cognitive tests' score, which indicates higher cognitive reserve of the elderly with higher educational levels. CONCLUSION: The functionality and independence of the so-called SuperAgers is determined by the cognitive reserve acquired throughout life, mainly developed by the years of formal education.

Depression is associated with self-rated frailty in older adults from an outpatient clinic: a prospective study.

ABSTRACTObjectives:The aim of the present study was to evaluate the association between depression and SSRI monotherapy and frailty both baseline and prospectively in older adults. DESIGN: Prospective cohort study, 12-month follow-up. SETTING: Geriatric outpatient clinic in São Paulo, Brazil. PARTICIPANTS: A total of 811 elderly adults aged 60 or older. MEASUREMENTS: Depression was diagnosed as follows: (1) a diagnosis of major depression disorder (MDD) according to DSM-5; or (2) an incomplete diagnosis of MDD, referred to as minor or subsyndromic depression, plus Geriatric Depression Scale 15-itens ≥ 6 points, and social or functional impairment secondary to depressive symptoms and observed by relatives. Frailty evaluation was performed through the FRAIL questionnaire, which is a self-rated scale. Trained investigators blinded to the baseline assessment conducted telephone calls to evaluate frailty after 12-month follow-up. The association between depression and the use of SSRI with frailty was estimated through a generalized estimating equation adjusted for age, gender, total drugs, and number of comorbidities. RESULTS: Depression with SSRI use was associated with frailty at baseline (OR 2.82, 95% CI = 1.69-4.69) and after 12 months (OR 2.75, 95% CI = 1.84-4.11). Additionally, depression with SSRI monotherapy was also associated with FRAIL subdomains Physical Performance (OR 1.99, 95% CI = 1.29-3.07) and Health Status (OR 4.64, 95% CI = 2.11-10.21). SSRI use, without significant depressive symptoms, was associated with subdomain Health Status (OR 1.52, 95% CI = 1.04-2.23). CONCLUSION: It appears that depression with SSRI is associated to frailty, and this association cannot be explained only by antidepressant use.

An integrated bioanalytical method development and validation approach: case studies.

We proposed an integrated bioanalytical method development and validation approach: (1) method screening based on analyte's physicochemical properties and metabolism information to determine the most appropriate extraction/analysis conditions; (2) preliminary stability evaluation using both quality control and incurred samples to establish sample collection, storage and processing conditions; (3) mock validation to examine method accuracy and precision and incurred sample reproducibility; and (4) method validation to confirm the results obtained during method development. This integrated approach was applied to the determination of compound I in rat plasma and compound II in rat and dog plasma. The effectiveness of the approach was demonstrated by the superior quality of three method validations: (1) a zero run failure rate; (2) >93% of quality control results within 10% of nominal values; and (3) 99% incurred sample within 9.2% of the original values. In addition, rat and dog plasma methods for compound II were successfully applied to analyze more than 900 plasma samples obtained from Investigational New Drug (IND) toxicology studies in rats and dogs with near perfect results: (1) a zero run failure rate; (2) excellent accuracy and precision for standards and quality controls; and (3) 98% incurred samples within 15% of the original values.

Effects of Adjuvant Chemotherapy on Cognitive Function of Patients With Early-stage Colorectal Cancer.

PURPOSE: Chemotherapy-related cognitive impairment can occur in cancer survivors after treatment, especially those patients who have undergone chemotherapy for breast cancer. The frequency and to what extent such toxicity develops in colorectal cancer (CRC) survivors is unknown. The present prospective study evaluated the effects of adjuvant chemotherapy on the cognitive performance of patients with localized CRC compared with a control group who had not undergone chemotherapy. PATIENTS AND METHODS: Consecutive patients with localized stage II and III CRC completed neuropsychological assessments, self-reported cognitive complaint questionnaires, and depressive symptom evaluations before starting fluoropyrimidine-based adjuvant chemotherapy and after 12 months. Blood was collected for apolipoprotein E genotyping. Diffusion tensor imaging data were acquired from a subset of participants at both evaluation points. RESULTS: From December 2012 to December 2014, 137 patients were approached and 85 were included. Of these 85 patients, 49 had undergone chemotherapy and 26 had not, in accordance with the standard recommendations for adjuvant therapy for CRC. The mean age was 62.5 ± 9.4 years, 60% were men, and the mean educational attainment was 7.6 ± 3.7 years. No difference was found in the global composite score (P = .38), attention (P = .84), or memory (P = .97) between the 2 groups during the follow-up period (mean ± standard deviation, 375 ± 29 days). However, a statistically significant difference was found for executive function after adjustment for age, sex, education, and depressive symptoms at baseline (ß -1.80; 95% confidence interval, -3.50 to -0.11; P = .04), suggesting worse performance for the chemotherapy group. For the 32 patients who had undergone magnetic resonance imaging, tract-based spatial statistics did not show voxelwise significant differences in structural brain connectivity at baseline or during follow-up. Apolipoprotein E polymorphisms were not predictive of cognitive dysfunction. CONCLUSION: Patients with CRC who received adjuvant 5-fluorouracil with or without oxaliplatin presented with a decline in executive function after 12 months compared with patients with localized disease who had not received chemotherapy.

Performance of the Pentagon Drawing test for the screening of older adults with Alzheimer's dementia.

The Pentagon Drawing Test (PDT) is a common cognitive screening test. OBJECTIVE: The aim of this study was to evaluate performance properties of a specific PDT scoring scale in older adults with Alzheimer's disease (AD) and healthy controls. METHODS: A cross-sectional study of 390 elderly patients, aged 60 years or older with at least two years of education was conducted. All participants completed clinical and neuropsychological evaluations, including the Cambridge Cognitive Examination, the Mini-Mental State Examination (MMSE), and the Clock Drawing Test. All PDT were blindly scored with the scale of Bourke et al. RESULTS: PDT analyses of the binary score on the MMSE (0 or 1 point) did not discriminate AD from controls (p = 0.839). However, when PDT was analyzed using the Bourke et al. scale, the two groups could be distinguished (p <0.001). PDT was not affected by education, showed sensitivity of 85.5% and specificity of 66.9%, discriminated different clinical stages of dementia, and correlated with the other cognitive tests (p <0.001). A 1-point difference on the Bourke et al. scale was associated with an odds ratio of 3.46 for AD. CONCLUSION: PDT can be used as a cognitive screen for suspected cases of dementia, especially AD, irrespective of educational level.

O teste do desenho do pentágono (PDT) é um teste de rastreio cognitivo simples. OBJETIVO: O objetivo deste estudo foi avaliar o desempenho de uma escala específica de pontuação da PDT em idosos com doença de Alzheimer (DA) e controles saudáveis. MÉTODOS: Estudo transversal, com 390 idosos, com mais de 60 anos de idade, com pelo menos dois anos de escolaridade. Todos os participantes passaram por anamnese clínica e neuropsicológica, incluindo o Cambridge Cognitive Examination (CAMCOG), o Mini-Exame do Estado Mental (MEEM) e o Teste do Desenho do Relógio (TDR). A avaliação do PDT com a escala Bourke et al. foi feita de forma cega. RESULTADOS: As análises PDT do escore binário do MEEM (0 e 1 ponto) não discriminaram DA dos controles (p = 0,839). Contudo, quando PDT foi avaliada pela escala Bourke et al., verificou-se diferenças estatisticamente significativa (p <0,001). A PDT não sofreu interferência da escolaridade, apresentando sensibilidade de 85,5% e especificidade de 66,9% para discriminar os diferentes estágios clínicos da demência. A escala também mostrou correlação com os testes cognitivos aplicados (p <0,001). Uma diferença de um ponto na escala Bourke et al. foi associada com OR (odds ratio) de 3,46 para DA. CONCLUSÃO: PDT pode ser utilizada como rastreio cognitivo para casos suspeitos de demência, especialmente DA, independentemente.

Accuracy of praxis test from Cambridge Cognitive Examination (CAMCOG) for Alzheimer's disease: a cross-sectional study.

BACKGROUND: Praxis impairment may be one of the first symptoms manifested in dementia, primarily in cortical dementia. The Cambridge Cognitive Examination (CAMCOG) evaluates praxis, but little is known about the accuracy of CAMCOG for diagnosing dementia. The aims here were to investigate the accuracy of praxis and its subitems in CAMCOG (constructive, ideomotor and ideational subitems) for diagnosing Alzheimer's disease (AD) among elderly patients. DESIGN AND SETTING: Cross-sectional study on community-dwelling elderly people. METHODS: 158 elderly patients were evaluated. CAMCOG, Mini-Mental State Examination and Pfeffer Functional Activities Questionnaire were used. ROC curve analysis was used to establish cutoff points. RESULTS: The total scores for praxis and the constructive subitem presented significant differences (P < 0.0001) between healthy elderly people and AD patients. Stage of dementia (clinical dementia rating, CDR = 0, 1 and 2) showed that total and constructive praxis can be used to classify the stages of dementia (mild and moderate cases), i.e. constructive praxis classified 88% of the patients with mild dementia (P < 0.0001) while total praxis classified 56% with moderate dementia. Comparison of normal controls (NC) and mild dementia cases showed specificity of 71% and sensitivity of 88% (AUC = 0.88; P < 0.0001). CONCLUSION: Some praxis subtests can have higher predictive diagnostic value for detecting Alzheimer's disease in mild stages (total praxis AUC = 0.858; P < 0.0001; constructive AUC = 0.972; P < 0.0001). Constructive praxis as measured using CAMCOG may contribute towards diagnosing dementia, because occurrence of impairment of praxis may help in recognizing an evolving dementia syndrome.

Use of 4ß-hydroxycholesterol in animal and human plasma samples as a biomarker for CYP3A induction.

BACKGROUND: 4ß-hydroxycholesterol (4ßHC) has recently been proposed as a potential endogenous biomarker for CYP3A activity. Developing bioanalytical assays for 4ßHC is challenging for several reasons, including endogenous background levels in plasma; the presence of free and ester forms; the inherent lack of MS sensitivity; and the presence of multiple positional isomers. RESULTS: Bioanalytical assays in mouse, rat, dog and human plasma were adapted and modified from a previous published human plasma assay for 4ßHC by using alkaline de-esterification, picolinic derivatization, a surrogate analyte (d7-4ßHC) in authentic matrices and chromatographic conditions that showed good separation from isobaric, positional isomers. CONCLUSION: These assays were applied to multiple studies and demonstrated potential applications of 4ßHC as a CYP3A biomarker across preclinical and clinical settings.

Accuracy of praxis test from Cambridge Cognitive Examination (CAMCOG) for Alzheimer's disease: a cross-sectional study

ABSTRACT BACKGROUND: Praxis impairment may be one of the first symptoms manifested in dementia, primarily in cortical dementia. The Cambridge Cognitive Examination (CAMCOG) evaluates praxis, but little is known about the accuracy of CAMCOG for diagnosing dementia. The aims here were to investigate the accuracy of praxis and its subitems in CAMCOG (constructive, ideomotor and ideational subitems) for diagnosing Alzheimer's disease (AD) among elderly patients. DESIGN AND SETTING: Cross-sectional study on community-dwelling elderly people. METHODS: 158 elderly patients were evaluated. CAMCOG, Mini-Mental State Examination and Pfeffer Functional Activities Questionnaire were used. ROC curve analysis was used to establish cutoff points. RESULTS: The total scores for praxis and the constructive subitem presented significant differences (P < 0.0001) between healthy elderly people and AD patients. Stage of dementia (clinical dementia rating, CDR = 0, 1 and 2) showed that total and constructive praxis can be used to classify the stages of dementia (mild and moderate cases), i.e. constructive praxis classified 88% of the patients with mild dementia (P < 0.0001) while total praxis classified 56% with moderate dementia. Comparison of normal controls (NC) and mild dementia cases showed specificity of 71% and sensitivity of 88% (AUC = 0.88; P < 0.0001). CONCLUSION: Some praxis subtests can have higher predictive diagnostic value for detecting Alzheimer's disease in mild stages (total praxis AUC = 0.858; P < 0.0001; constructive AUC = 0.972; P < 0.0001). Constructive praxis as measured using CAMCOG may contribute towards diagnosing dementia, because occurrence of impairment of praxis may help in recognizing an evolving dementia syndrome.

Performance of the Pentagon Drawing test for the screening of older adults with Alzheimer's dementia / Desempenho do teste do desenho do pentágono para rastreio de idosos com demência de alzheimer

ABSTRACT. The Pentagon Drawing Test (PDT) is a common cognitive screening test. Objective: The aim of this study was to evaluate performance properties of a specific PDT scoring scale in older adults with Alzheimer's disease (AD) and healthy controls. Methods: A cross-sectional study of 390 elderly patients, aged 60 years or older with at least two years of education was conducted. All participants completed clinical and neuropsychological evaluations, including the Cambridge Cognitive Examination, the Mini-Mental State Examination (MMSE), and the Clock Drawing Test. All PDT were blindly scored with the scale of Bourke et al. Results: PDT analyses of the binary score on the MMSE (0 or 1 point) did not discriminate AD from controls (p = 0.839). However, when PDT was analyzed using the Bourke et al. scale, the two groups could be distinguished (p <0.001). PDT was not affected by education, showed sensitivity of 85.5% and specificity of 66.9%, discriminated different clinical stages of dementia, and correlated with the other cognitive tests (p <0.001). A 1-point difference on the Bourke et al. scale was associated with an odds ratio of 3.46 for AD. Conclusion: PDT can be used as a cognitive screen for suspected cases of dementia, especially AD, irrespective of educational level.

RESUMO. O teste do desenho do pentágono (PDT) é um teste de rastreio cognitivo simples. Objetivo: O objetivo deste estudo foi avaliar o desempenho de uma escala específica de pontuação da PDT em idosos com doença de Alzheimer (DA) e controles saudáveis. Métodos: Estudo transversal, com 390 idosos, com mais de 60 anos de idade, com pelo menos dois anos de escolaridade. Todos os participantes passaram por anamnese clínica e neuropsicológica, incluindo o Cambridge Cognitive Examination (CAMCOG), o Mini-Exame do Estado Mental (MEEM) e o Teste do Desenho do Relógio (TDR). A avaliação do PDT com a escala Bourke et al. foi feita de forma cega. Resultados: As análises PDT do escore binário do MEEM (0 e 1 ponto) não discriminaram DA dos controles (p = 0,839). Contudo, quando PDT foi avaliada pela escala Bourke et al., verificou-se diferenças estatisticamente significativa (p <0,001). A PDT não sofreu interferência da escolaridade, apresentando sensibilidade de 85,5% e especificidade de 66,9% para discriminar os diferentes estágios clínicos da demência. A escala também mostrou correlação com os testes cognitivos aplicados (p <0,001). Uma diferença de um ponto na escala Bourke et al. foi associada com OR (odds ratio) de 3,46 para DA. Conclusão: PDT pode ser utilizada como rastreio cognitivo para casos suspeitos de demência, especialmente DA, independentemente.