Bioanalytical HPLC Applications of In-Tube Solid Phase Microextraction: A Two-Decade Overview.

In-tube solid phase microextraction is a cutting-edge sample treatment technique offering significant advantages in terms of miniaturization, green character, automation, and preconcentration prior to analysis. During the past years, there has been a considerable increase in the reported publications, as well as in the research groups focusing their activities on this technique. In the present review article, HPLC bioanalytical applications of in-tube SPME are discussed, covering a wide time frame of twenty years of research reports. Instrumental aspects towards the coupling of in-tube SPME and HPLC are also discussed, and detailed information on materials/coatings and applications in biological samples are provided.

Validation of the Spartan RXCYP2C19 Genotyping Assay Utilizing Blood Samples.

The antiplatelet agent clopidogrel, a prodrug that requires bioactivation through the cytochrome P450 2C19 (CYP2C19) enzyme, is commonly prescribed post-percutaneous coronary intervention (PCI). Genetic variation in CYP2C19 contributes to individual variability in clopidogrel response, and can lead to adverse cardiovascular events. Incorporating CYP2C19 testing during routine clinical care helps identify high-risk patients, and provides the opportunity for pharmacotherapeutic interventions in the early post-PCI period. The Spartan RX CYP2C19 System has emerged as an optimal genotyping assay for use in clinical care due to ease of use, utilization of buccal swabs, and rapid turnaround time. However, workflow constraints related to sample collection and processing, storage, time, and personnel were encountered when integrating testing into clinical care. To improve clinical workflow and successfully implement CYP2C19 genotyping at our institution, we validated the Spartan RX System to return genotype utilizing blood samples. Our Molecular Diagnostic Laboratory tested 26 known reference materials and both blood and buccal swab samples from 23 patients and volunteers using the Spartan RX Assay. Genotype results were 100% concordant between DNA from blood and buccal swabs for all patients or volunteers, and consistent with expected results for the 26 reference materials. For reproducibility, three samples were tested in at least four separate runs, with all resulting genotypes in agreement between runs. Post-validation, the laboratory began offering CYP2C19 testing during clinical care. DNA extracted from blood can serve as a genomic DNA source for the Spartan RX Assay. Alteration of the methodology allowed for clinical implementation to support genotype-guided therapy.

Emerging Role of Organ-on-a-Chip Technologies in Quantitative Clinical Pharmacology Evaluation.

The recently enacted Prescription Drug User Fee Act (PDUFA) VI includes in its performance goals "enhancing regulatory science and expediting drug development." The key elements in "enhancing regulatory decision tools to support drug development and review" include "advancing model-informed drug development (MIDD)." This paper describes (i) the US Food and Drug Administration (FDA) Office of Clinical Pharmacology's continuing efforts in developing quantitative clinical pharmacology models (disease, drug, and clinical trial models) to advance MIDD, (ii) how emerging novel tools, such as organ-on-a-chip technologies or microphysiological systems, can provide new insights into physiology and disease mechanisms, biomarker identification and evaluation, and elucidation of mechanisms of adverse drug reactions, and (iii) how the single organ or linked organ microphysiological systems can provide critical system parameters for improved physiologically-based pharmacokinetic and pharmacodynamic evaluations. Continuous public-private partnerships are critical to advance this field and in the application of these new technologies in drug development and regulatory review.

Essential Medications for Patients With Suspected or Confirmed Ebola Virus Disease in Resource-Limited Environments.

BACKGROUND: In 2014, the U.S. Public Health Service (USPHS) Commissioned Corps deployed to Monrovia, Liberia, to operate a 25-bed Ebola treatment unit (ETU) constructed by the U.S. Military. The ETU was named the Monrovia Medical Unit (MMU) and was constructed from an U.S. Air Force Expeditionary Medical Support (EMEDS) unit with modifications on the basis of consultation from Médecins Sans Frontières, the World Health Organization, and expert panels from the U.S. Department of Defense and Department of Health and Human Services. From November 12, 2014, to April 30, 2015, 42 patients (18 confirmed Ebola virus disease [EVD] and 24 suspected EVD) from nine countries were treated by USPHS providers at the MMU. The medications used in the MMU were primarily procured from the EMEDS 25-bed pharmacy cache. However, specific formulary additions were made for treatment of EVD. METHODS: Using the MMU pharmacy dispensing data, we compared and contrasted the medications used in the MMU with recommendations in published EVD treatment guidelines for austere settings. FINDINGS: After comparing and contrasting the MMU pharmacy dispensing data with publications with EVD medication recommendations applicable to resource-limited settings, 101 medications were included in the USPHS Essential Medications for the Management of EVD List (EML) for an austere, isolated clinical environment. DISCUSSION/IMPACT/RECOMMENDATIONS: Because Ebola outbreaks often occur in remote areas, proactive planning, improved preparedness, and optimal patient care for EVD are needed, especially in the context of austere environments with a scarcity of resources. We developed the EML to assist in the planning for future Ebola outbreaks in a remote clinical environment and to provide a list of medications that have been used in an ETU. The EML is a comprehensive medication list that builds on the existing publications with EVD treatment recommendations applicable to supply-constrained clinical environments. As well, it is a resource for the provision of medications when evaluating donations, procurement, and may help inform estimates for product inventory requirements for an ETU. We hope the EML will improve readiness and enhance the capabilities of local and regional international responders.

PhID: An Open-Access Integrated Pharmacology Interactions Database for Drugs, Targets, Diseases, Genes, Side-Effects, and Pathways.

The current network pharmacology study encountered a bottleneck with a lot of public data scattered in different databases. There is a lack of an open-access and consolidated platform that integrates this information for systemic research. To address this issue, we have developed PhID, an integrated pharmacology database which integrates >400 000 pharmacology elements (drug, target, disease, gene, side-effect, and pathway) and >200 000 element interactions in branches of public databases. PhID has three major applications: (1) assisting scientists searching through the overwhelming amount of pharmacology element interaction data by names, public IDs, molecule structures, or molecular substructures; (2) helping visualizing pharmacology elements and their interactions with a web-based network graph; and (3) providing prediction of drug-target interactions through two modules: PreDPI-ki and FIM, by which users can predict drug-target interactions of PhID entities or some drug-target pairs of their own interest. To get a systems-level understanding of drug action and disease complexity, PhID as a network pharmacology tool was established from the perspective of data layer, visualization layer, and prediction model layer to present information untapped by current databases.

Integrating clinical metabolomics-based biomarker discovery and clinical pharmacology to enable precision medicine.

Novel developments in biomarkers discovery are essential in modern health care, notably in treatment individualization and precision medicine. Clinical metabolomics, which aims to identify small molecule metabolites present in patient-derived samples, has attracted much attention to support discovery of novel biomarkers. However, the step from discriminatory features of disease states towards biomarkers that can truly individualize treatments is challenging. Biomarkers used for treatment individualization can either be dynamic or static prognostic biomarkers. Dynamic biomarkers are relevant for describing the clinical response, including dynamical disease progression and associated treatment response. Static (prognostic) biomarkers do not describe but rather predict a clinical response, and typically reflect aspects of the physiological state of a patient related to drug treatment response or disease progression dynamics. Pharmacokinetic-pharmacodynamic (PK-PD) modeling represents an established approach for drug treatment individualization based on drug exposure or treatment response biomarkers, as well as for the description of disease progression dynamics. Here, we discuss how novel treatment individualization biomarkers can be identified using a clinical metabolomics-based approach, and how concepts inspired from the field of PK-PD modeling can be integrated in this process in order to increase the clinical relevance of identified biomarkers and precision medicine.

Tratamiento farmacológico para la Osteoporosis y su influencia en el proceso de consolidación de las fracturas / Pharmacologic treatment for osteoporosis and its influence on fracture healing

Las fracturas por fragilidad o secundarias a la osteoporosis se pueden considerar como un importante problema de Salud Pública por las consecuencias en términos de mortalidad y morbilidad que generan. Y las previsiones para el futuro, teniendo en cuenta el progresivo envejecimiento de la población y el aumento de la esperanza de vida, no son nada optimistas. Suponen y supondrán un importante consumo de recursos. Una de las estrategias que ha demostrado eficacia para su prevención, es el tratamiento farmacológico. Pero este tratamiento puede afectar de una manera u otra al proceso de consolidación, uno de los objetivos primarios fundamentales en su manejo. En este trabajo de revisión queremos establecer, en base a la evidencia actual, cómo afectan todos y cada uno de los fármacos indicados para el tratamiento de la osteoporosis al proceso de consolidación de las fracturas por fragilidad, una cuestión que genera dudas e incertidumbres en el traumatólogo, cuya responsabilidad y papel en el escenario de la prevención, es fundamental

Osteoporotic fragility fractures can be seen as a major public health problem because their consequences in terms of mortality and morbidity. Taking into account the progressive ageing of the population and the increase in life expectancy, the expectative in the next future are not optimistic. They are and will be an important focus of health resources consumption. One of the strategies that have proven be effective for fracture prevention, is the antiosteoporotic pharmacological treatment. This type of treatments can affect bone healing process in one way or another. In this review, based on the current scientific evidence, we want to establish how each one of the drugs prescribed for the osteoporosis treatment affects the bone healing process of fragility fractures, an issue that generates doubts and uncertainties in the orthopaedic surgeon, whom responsibility and role in fracture prevention is essential

Smartphone apps to support hospital prescribing and pharmacology education: a review of current provision.

Junior doctors write the majority of hospital prescriptions but many indicate they feel underprepared to assume this responsibility and around 10% of prescriptions contain errors. Medical smartphone apps are now widely used in clinical practice and present an opportunity to provide support to inexperienced prescribers. This study assesses the contemporary range of smartphone apps with prescribing or related content. Six smartphone app stores were searched for apps aimed at the healthcare professional with drug, pharmacology or prescribing content. Three hundred and six apps were identified. 34% appeared to be for use within the clinical environment in order to aid prescribing, 14% out with the clinical setting and 51% of apps were deemed appropriate for both clinical and non-clinical use. Apps with drug reference material, such as textbooks, manuals or medical apps with drug information were the commonest apps found (51%), followed by apps offering drug or infusion rate dose calculation (26%). 68% of apps charged for download, with a mean price of £14.25 per app and a range of £0.62-101.90. A diverse range of pharmacology-themed apps are available and there is further potential for the development of contemporary apps to improve prescribing performance. Personalized app stores may help universities/healthcare organizations offer high quality apps to students to aid in pharmacology education. Users of prescribing apps must be aware of the lack of information regarding the medical expertise of app developers. This will enable them to make informed choices about the use of such apps in their clinical practice.

Diseño de un instrumento para evaluar los factores que influyen en la adherencia a tratamientos, en personas que presentan factores de riesgo de enfermedad cardiovascular / Design of an instrument to evaluate the factors influencing the treatment adherence in people with cardiovascular disease risks

El propósito de este estudio fue diseñar y probar la validez de contenido de un instrumento creado para evaluar los factores que influyen en la adherencia a tratamientos farmacológicos y no farmacológicos en personas que presenten factores de riesgo de enfermedad cardiovascular. Se creó este instrumento y se sometió al concepto de nueve enfermeras expertas, seleccionadas por su experiencia en el área de cuidado a este tipo de pacientes, por su nivel académico y su interés en la investigación. Se utilizó el método de Fehring para establecer validez de contenido que se ha utilizado en enfermería por la North American Nursing Diagnosis Association (NANDA) y por el proyecto de la Universidad de Iowa para validar resultados e intervenciones en la taxonomía. La validez de contenido se estableció desde tres perspectivas: validez de contenido, validez según ítems y dimensiones y validez de contenido general. Los resultados de validez según los expertos y los de validez general mostraron un índice de validez de contenido de 0.91. La validez según los expertos indicó que los ítems y el total del contenido del instrumento son representativos del concepto que se desea medir y que son claros.

The purpose of this study was to design and verify the validity of content and the apparent validity of a tool created to assess the factors with an influence on the adherence to pharmacologic and non pharmacologic treatments in people with a risk for cardiovascular diseases. This tool was created and submitted for concept to 9 proficient nurses chosen for their experience in this patient care area, for their academic level and their interest for research. The Fehring method was used to establish validity for the content which has been used in nursing by the North American Nursing Diagnosis Association (NANDA) and by the University of Iowa’s Project to validate results and taxonomy interventions. The content validity was established from three perspectives; content validity, validity...

Application of a static fluorescence-based cytometer (the CellScan) in basic cytometric studies, clinical pharmacology, oncology and clinical immunology.

The CellScan apparatus is a laser scanning cytometer enabling repetitive fluorescence intensity (FI) and polarization (FP) measurements in living cells, as a means of monitoring lymphocyte activation. The CellScan may serve as a tool for diagnosis of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) as well as other autoimmune diseases by monitoring FP changes in peripheral blood lymphocytes (PBLs) following exposure to autoantigenic stimuli. Changes in FI and FP in atherosclerotic patients' PBLs following exposure to various stimuli have established the role of the immune system in atherosclerotic disease. The CellScan has been evaluated as a diagnostic tool for drug-allergy, based on FP reduction in PBLs following incubation with allergenic drugs. FI and FP changes in cancer cells have been found to be well correlated with the cytotoxic effect of anti-neoplastic drugs. In conclusion, the CellScan has a variety of applications in cell biology, immunology, cancer research and clinical pharmacology.

Monitoring of idebenone treatment in patients with Friedreich's ataxia by high-pressure liquid chromatography with electrochemical detection.

Idebenone is a quinone analog that is applied in the treatment of several neurological disorders including Friedreich ataxia and mitochondrial encephalomyopathies. Our aim was to develop an easy and sensitive analytical HPLC-procedure for the determination of idebenone in the serum of patients treated with this drug. Serum samples from nine paediatric patients diagnosed with Friedreich ataxia and receiving idebenone treatment were analyzed. Idebenone was separated from serum by reverse high-pressure liquid chromatography and analyzed using an electrochemical detection procedure. No interferences were observed during analysis of patient samples obtained prior to idebenone treatment. Calibration of idebenone concentration indicated a linear range between 500 pmol/l and 5 micromol/l and calculation of within-run and between-run coefficients of variation suggested adequate analytical quality for reliable determination. In agreement with previously reported data, during drug therapy, idebenone serum concentrations (basal conditions, range 0.1-0.49 micromol/l) were greatly elevated 90 min after an oral dose (range 0.66-3.63 micromol/l). Thus, we have developed a simple and rapid method that offers adequate analytical quality for accurate idebenone determination.

Recent trends in laboratory automation in the pharmaceutical industry.

The impact of robotics and automation on the pharmaceutical industry over the last two decades has been significant. In the last ten years, the emphasis of laboratory automation has shifted from the support of manufactured products and quality control of laboratory applications, to research and development. This shift has been the direct result of an increased emphasis on the identification, development and eventual marketing of innovative new products. In this article, we will briefly identify and discuss some of the current trends in laboratory automation in the pharmaceutical industry as they apply to research and development, including screening, sample management, combinatorial chemistry, ADME/Tox and pharmacokinetics.

Positron emission tomography: from production and distribution to drug research and clinical applications.

The radiopharmaceutical FDG has had a major impact on PET imaging in clinical medicine, particularly in the detection and staging of certain cancers. PET isotopes offer unparalleled insight into in vivo distribution of labeled drugs. PET imaging of blood flow, metabolism, and neuroreceptor characteristics may provide new perspectives on the mechanism of action and effects of drugs.

Polysomnography in drug development.

Polysomnography in drug development is used to detect desirable and undesirable effects of drugs on normal and disturbed sleep. Although this method is essential for the approval of new hypnotic drugs, it is quite often neglected in the development of drugs that show unwanted side effects on normal sleep. In this review, the requirements for qualified polysomnography are described, the strong and weak points of the method are discussed, and its importance for the drug development process is pointed out.

Effect of educational level, gender and age on the performance in a multi-user computerized psychometric test system for use in clinical pharmacological studies.

197 native, drug-free healthy young volunteers performed psychometric tests on a single occasion, using a new multi-user computerized test system which consists of tests of simple reaction time, complex reaction time, vigilance, concentration, motor coordination, short-term memory (word pairs or figures) and abstract language-free reasoning (2 versions). Normality of distribution of all psychometric variables was checked. For the reasoning tests and the memory tests, internal consistency and parallel test reliability were determined. Cross-correlations between the variables and factor analysis were done to evaluate whether different tests measure different brain functions. Multivariate variance analysis was carried out to test the effect of the independent factors school education, gender and age on the performance in the psychometric tests. Subjects with a lower school education level performed worse in the reasoning tests, the concentration test and the memory tests. Females were slower in the coordination test and made fewer correct solutions in the concentration test. Older subjects performed worse in the reasoning tests and had a longer working time in the memory tests than younger ones. The results show the necessity of psychometric screening of volunteers before recruitment for clinical pharmacological psychometric studies in order to reduce-individual variability.