Assessing the overall medication use by elderly people in a Brazilian hospital using the start/stopp criteria version 2

To estimate the frequency of the use of medicines listed in the Screening Tool to Alert Doctors to the Right Treatment (START) and Screening Tool of Older Person's Prescriptions (STOPP) criteria version 2 among the elderly. A cross-sectional study was conducted on elderly who were attended in medical clinic and cardiology sectors in a hospital in southern Brazil attended at a hospital from February through September 2016. A data-collection tool was used to obtain information on variables, such as demographic and clinical data, and medications used before and during the hospitalization period. The adequacy of the medicines taken was examined with regard to omission (START) or inappropriate use (STOPP). This study was approved by the Research Ethics Committee of the University of Southern Santa Catarina. A total of 307 subjects were included in the final sample. The mean age was 75.2 years (SD = 8; range 65-102). Of the total, 93.5% had had at least one potential prescribing omission (PPO) listed in the START criteria, whereas 95.4% used at least one medicine of the STOPP criteria. PPO was significantly associated with lower mean age (74.9 years, SD = 7.9 versus 79.0 years, SD = 8.8) among the elderly who did not have PPOs detected by the START criteria (p-value=0.03). Furthermore, PPO was associated with longer hospital stay (18 versus 9 days; p-value=0.03). This study revealed inadequate prescription affecting 99.3% of the participating patients. To the best of our knowledge, this was the first to use the START and STOPP criteria, version 2, in Brazil.

Plasmonic Au nanostar Raman probes coupling with highly ordered TiO2/Au nanotube arrays as the reliable SERS sensing platform for chronic myeloid leukemia drug evaluation.

The accurate therapeutic evaluation for chronic myeloid leukemia (CML) drug is of great importance to minimize side effects and enhance efficacy. Herein, a facile and precise surface-enhanced scattering (SERS) approach based on coupled plasmonic field has been introduced to evaluate the therapeutic outcomes of antileukemia drug through ultrasensitive assay of caspase-3 activity in apoptotic cells. Caspase-3 as an apoptosis indicator could specifically cleave the N-terminus of biotinylated DEVD-peptide (biotin-Gly-Asp-Gly-Asp-Glu-Val-Asp-Gly-Cys) immobilized on the Au nanoparticle-decorated TiO2 nanotube arrays (TiO2/Au NTAs) substrate. After the enzyme cleavage with caspase-3, Raman-labelled Au nanostar (AuNS) probes captured the residual DEVD-peptides via the recognition between streptavidin and biotin, thus resulting in an enhanced Raman response on the SERS platform. The variation of Raman intensity revealed caspase-3 activity that reflected the chemotherapeutic effect. On this platform, AuNS nanoprobes offered a large number of binding sites and intrinsic "hot spots" for Raman reporters, while TiO2/Au NTAs rendered a homogenously coupled electromagnetic field between the adjacent repeated units over the large area. In particular, a spatially expanding plasmonic field formed by coupling AuNSs with TiO2/Au NTAs would further heighten Raman enhancement. Taking these advantages, the strong and uniform Raman signals were achieved. Furthermore, the practicability investigation witnessed that the proposed SERS strategy was available to evaluate the therapeutic effect of dasatinib on CML K562 cells. The developed method possesses fascinating advantages of cost-effectiveness, excellent reproducibility and high sensitivity, which endows it with promising potential in apoptosis monitoring and anticancer drug development.

Organs-on-Chips: How Microsystems Technology Can Transform the Drug Development Process.

The drug development pipeline, once one of the most successful and lucrative commercial sectors in the United States, is now strained by a combination of factors: increased development costs, lengthy time lines, and the poor predictive power of preclinical studies, among others. These factors, in combination with the need to respond to newly evolving demands?including the trend toward personalized or precision medicine, rising rates for many chronic diseases, and continued threats from emerging infectious diseases?are placing extraordinary pressure on an already strained development process.

Design and development of a robotized system coupled to µCT imaging for intratumoral drug evaluation in a HCC mouse model.

Hepatocellular carcinoma (HCC) is one of the most common cancer related deaths worldwide. One of the main challenges in cancer treatment is drug delivery to target cancer cells specifically. Preclinical evaluation of intratumoral drugs in orthotopic liver cancer mouse models is difficult, as percutaneous injection hardly can be precisely performed manually. In the present study we have characterized a hepatoma model developing a single tumor nodule by implantation of Hep55.1C cells in the liver of syngeneic C57BL/6J mice. Tumor evolution was followed up by µCT imaging, and at the histological and molecular levels. This orthotopic, poorly differentiated mouse HCC model expressing fibrosis, inflammation and cancer markers was used to assess the efficacy of drugs. We took advantage of the high precision of a previously developed robotized system for automated, image-guided intratumoral needle insertion, to administer every week in the tumor of the Hep55.1C mouse model. A significant tumor growth inhibition was observed using our robotized system, whereas manual intraperitoneal administration had no effect, by comparison to untreated control mice.

Three-dimensional in vitro tumor models for cancer research and drug evaluation.

Cancer occurs when cells acquire genomic instability and inflammation, produce abnormal levels of epigenetic factors/proteins and tumor suppressors, reprogram the energy metabolism and evade immune destruction, leading to the disruption of cell cycle/normal growth. An early event in carcinogenesis is loss of polarity and detachment from the natural basement membrane, allowing cells to form distinct three-dimensional (3D) structures that interact with each other and with the surrounding microenvironment. Although valuable information has been accumulated from traditional in vitro studies in which cells are grown on flat and hard plastic surfaces (2D culture), this culture condition does not reflect the essential features of tumor tissues. Further, fundamental understanding of cancer metastasis cannot be obtained readily from 2D studies because they lack the complex and dynamic cell-cell communications and cell-matrix interactions that occur during cancer metastasis. These shortcomings, along with lack of spatial depth and cell connectivity, limit the applicability of 2D cultures to accurate testing of pharmacologically active compounds, free or sequestered in nanoparticles. To recapitulate features of native tumor microenvironments, various biomimetic 3D tumor models have been developed to incorporate cancer and stromal cells, relevant matrix components, and biochemical and biophysical cues, into one spatially and temporally integrated system. In this article, we review recent advances in creating 3D tumor models employing tissue engineering principles. We then evaluate the utilities of these novel models for the testing of anticancer drugs and their delivery systems. We highlight the profound differences in responses from 3D in vitro tumors and conventional monolayer cultures. Overall, strategic integration of biological principles and engineering approaches will both improve understanding of tumor progression and invasion and support discovery of more personalized first line treatments for cancer patients.

Engineering and evaluating drug delivery particles in microfluidic devices.

The development of new and improved particle-based drug delivery is underpinned by an enhanced ability to engineer particles with high fidelity and integrity, as well as increased knowledge of their biological performance. Microfluidics can facilitate these processes through the engineering of spatiotemporally highly controlled environments using designed microstructures in combination with physical phenomena present at the microscale. In this review, we discuss microfluidics in the context of addressing key challenges in particle-based drug delivery. We provide an overview of how microfluidic devices can: (i) be employed to engineer particles, by providing highly controlled interfaces, and (ii) be used to establish dynamic in vitro models that mimic in vivo environments for studying the biological behavior of engineered particles. Finally, we discuss how the flexible and modular nature of microfluidic devices provides opportunities to create increasingly realistic models of the in vivo milieu (including multi-cell, multi-tissue and even multi-organ devices), and how ongoing developments toward commercialization of microfluidic tools are opening up new opportunities for the engineering and evaluation of drug delivery particles.

A microfluidic method to measure small molecule diffusion in hydrogels.

Drug release from a fluid-contacting biomaterial is simulated using a microfluidic device with a channel defined by solute-loaded hydrogel; as water is pumped through the channel, solute transfers from the hydrogel into the water. Optical analysis of in-situ hydrogels, characterization of the microfluidic device effluent, and NMR methods were used to find diffusion coefficients of several dyes (model drugs) in poly(ethylene glycol) diacrylate (PEG-DA) hydrogels. Diffusion coefficients for methylene blue and sulforhodamine 101 in PEG-DA calculated using the three methods are in good agreement; both dyes are mobile in the hydrogel and elute from the hydrogel at the aqueous channel interface. However, the dye acid blue 22 deviates from typical diffusion behavior and does not release as expected from the hydrogel. Importantly, only the microfluidic method is capable of detecting this behavior. Characterizing solute diffusion with a combination of NMR, optical and effluent methods offer greater insight into molecular diffusion in hydrogels than employing each technique individually. The NMR method made precise measurements for solute diffusion in all cases. The microfluidic optical method was effective for visualizing diffusion of the optically active solutes. The optical and effluent methods show potential to be used to screen solutes to determine if they elute from a hydrogel in contact with flowing fluid. Our data suggest that when designing a drug delivery device, analyzing the diffusion from the molecular level to the device level is important to establish a complete picture of drug elution, and microfluidic methods to study such diffusion can play a key role.

Laser diode thermal desorption-positive mode atmospheric pressure chemical ionization tandem mass spectrometry for the ultra-fast quantification of a pharmaceutical compound in human plasma.

An ultra-fast, reliable and sensitive analytical method enabling high-throughput quantitative analysis of pharmaceutical compounds in human plasma is described. The quantitative work was performed on one of our compound currently under clinical trial by employing a deuterated internal standard (IS). Plasma samples were treated on solid phase micro-extraction (SPME) plates prior their analysis by laser diode thermal desorption and atmospheric pressure chemical ionization coupled to tandem mass spectrometry (LDTD/APCI-MS/MS) in positive mode. The sample analysis run time was 10s as compared to the 7 min obtained for the validated LC-MS/MS method. The limit of quantification (LOQ) of the method was estimated at 1 ng/mL. The calibration graphs were linear with a regression coefficient R(2) > 0.997. The data of the partial validation show that LDTD/APCI-MS/MS assay was highly reproducible and selective. In addition, the deviations for intra and inter assay accuracy and precision data were within 15% at all quality control levels. The LDTD/APCI-MS/MS method was successfully applied to the analysis of clinical samples and the data obtained were consistent with those found with a validated LC-MS/MS assay. This work demonstrates that LDTD/APCI-MS/MS could be used for the ultra-fast and reliable quantitative analysis of pharmaceutical compounds in human plasma without using the separation step commonly associated with the LC-MS/MS assay.

Studying drug-plasma protein interactions by two-injector microchip electrophoresis frontal analysis.

We developed a simple, rapid, and sensitive two-injector microchip electrophoresis frontal analysis (MCE-FA) method for studying drug-plasma protein interactions. In this method, large volumes of a reference sample and drug-plasma protein mixture were simultaneously introduced into the respective sections of the microchannel through the separated injectors and then electrophoresed. Since the reference sample did not meet with the interacting species during migration, it could be used as an external standard. The interaction between heparin and HSA was quantitatively characterized as a model system. The binding constant was found to be (1.53 +/- 0.01) x 10(4) M(-1).

Learning from the data: mining of large high-throughput screening databases.

High-throughput screening (HTS) campaigns in pharmaceutical companies have accumulated a large amount of data for several million compounds over a couple of hundred assays. Despite the general awareness that rich information is hidden inside the vast amount of data, little has been reported for a systematic data mining method that can reliably extract relevant knowledge of interest for chemists and biologists. We developed a data mining approach based on an algorithm called ontology-based pattern identification (OPI) and applied it to our in-house HTS database. We identified nearly 1500 scaffold families with statistically significant structure-HTS activity profile relationships. Among them, dozens of scaffolds were characterized as leading to artifactual results stemming from the screening technology employed, such as assay format and/or readout. Four types of compound scaffolds can be characterized based on this data mining effort: tumor cytotoxic, general toxic, potential reporter gene assay artifact, and target family specific. The OPI-based data mining approach can reliably identify compounds that are not only structurally similar but also share statistically significant biological activity profiles. Statistical tests such as Kruskal-Wallis test and analysis of variance (ANOVA) can then be applied to the discovered scaffolds for effective assignment of relevant biological information. The scaffolds identified by our HTS data mining efforts are an invaluable resource for designing SAR-robust diversity libraries, generating in silico biological annotations of compounds on a scaffold basis, and providing novel target family specific scaffolds for focused compound library design.

Evaluación de la película lagrimal con métodos diagnósticos invasivos vs método diagnóstico no invasivo / Invasive Diagnostic Methods vs. Non-Invasive Diagnostic Methods in the Evaluation of Tear Film

Debido a la gran importancia fisiológica y óptica que representa la película lagrimal para el correcto funcionamiento del ojo humano, una acertada evaluación y diagnostico de cualquier tipo de anomalía o alteración, ya sea en sus mecanismos de secreción, estabilidad o calidad, de una o todas sus capas, se convierte en una imperiosa necesidad para la práctica optométrica y oftalmológica. La evaluación de la película preocular lagrimal (PLPO) es de gran ayuda diagnóstica en salud visual. En muchos países se vienen empleando técnicas no invasivas, basadas en interferometría 1, 2, 3; las técnicas invasivas (BUT) tiempo de rompimiento de la película lagrimal, han demostrado no tener validez significativa por su baja reproducibilidad 4,5 y por provocar una alteración bioquímica de la película lagrimal, inducida por la fluoresceína. Este estudio se llevó a cabo con el fin de evaluar una técnica de rompimiento de la película lagrimal no invasiva (BUTNI) con Tearscope® de Keeler U.K., basada en principios físicos de interferometría, a través de colores de interferencia generados por las diferentes longitudes de onda 6, 7, 8. Este instrumento mide el espesor de la capa lipídica y el reservorio lagrimal, e igualmente permite observar la calidad de la película lagrimal. Se escogió una muestra de 60 sujetos presumiblemente sanos (n= 60), de acuerdo con los criterios de inclusión y exclusión y por medio del cuestionario validado McMonnies 9, 10, 11, para diagnóstico de ojo seco, se evaluaron ambos ojos. Con el fin de descartar alteraciones en la visión del color, los examinadores fueron sometidos a la prueba Farnsworth D1512 para evitar sesgos en la observación de los patrones de colores con el Tearscope. En una primera sesión se evaluaron el BUTNI/h 7,/h 8, BUT con fluoresceína al 2 por cien (FulGlo) y Schirmer Test I13, 14, 15 y en la segunda sesión Schirmer Test II con proximetacaína al 0,5 por cien, para evitar la interacción farmacológica entre proximetacaina y fluore...

Issues in drug delivery: concepts and practice.

Understanding the transport and deposition of inhaled aerosols is of fundamental importance to inhalation therapy. Herein we address issues that affect drug delivery from experimental and theoretical perspectives. Accordingly, we shall limit our comments to a focused review of laboratory work (ie, an in vitro perspective) and the development of a computer-based 3-dimensional (3D) oral morphology with related computational fluid dynamics (CFD) and particle deposition studies (ie, an "in silico" perspective). To describe the oral region, morphometric data from the literature were employed. With Maya Unlimited, a third-party animation software package, coronal images were used to create initial spline curves, which served as the foundation of a nonuniform rational B-spline surface, representing a 3D morphology. To the best of our knowledge, this study is the first medical application of Maya Unlimited. We have demonstrated that the code can be employed to construct 3D biological structures and perform 3D CFD simulations of aerosols from dry powder inhalers and metered-dose inhalers. A study was also conducted using Fluent, a commercially available software package that has been used extensively in our laboratory for 3D CFD computations. The Maya Unlimited software can generate physiologically realistic oral structures; it has great potential for use in the medical arena, because it requires neither advance technical training nor substantial peripheral ( eg, hardware) support, it allows for the introduction of medical devices ( eg, dry powder inhalers) into simulations, and it predicts 3D CFDpatterns consistent with experimental observations and results of more rigorous software ( Fluent). In the in vitro perspective we considered numerous salient topics, including the performances of dry powder inhalers and metered-dose inhalers, their respective operating characteristics, and relevance to in vivo data. We advocate that 3D CFD software be employed in a complementary manner, in real time, with aerosol therapy protocols in the medical arena, to promote the targeted delivery of inhaled drugs and thereby enhance their efficacies.

The advantages of the Ussing chamber in drug absorption studies.

By adding high concentrations of test drugs to an Ussing chamber with rat jejunum, we established a system that yields very high correlations between the rat absorption percentage and the membrane permeability, and that can accurately predict the absorption percentage for rats. An advantage of this technique is that, unlike the results obtained using Caco-2, the slope of the absorption/membrane-permeability curve is gentle, which facilitates a more exact prediction of the absorption percentage. In addition, the results obtained with this technique demonstrated that it could be used to evaluate the absorption percentage of drugs with an affinity for P-glycoprotein (P-gp), which cannot be assessed using Caco-2. This method also allows for cassette screening, which would facilitate evaluation of the contribution of P-gp to absorption in the small intestine. Cassette screening showed that absorption of fexofenadine was unaffected by combination with the P-gp substrate ketoconazole. Consistent with this finding, in vivo studies showed that ketoconazole did not affect the Fa Fg for fexofenadine, a pharmacokinetic parameter that reflects absorption and bioavailability in the small intestine. This confirms the usefulness of the Ussing chamber for cassette screening and also suggests that intestinal P-gp has a minimal contribution to drug absorption.

Fully automated solid weighing workstation.

A fully automated, solid-to-solid weighing workstation (patent pending) is described in this article. The core of this automated process is the use of an electrostatically charged pipette tip to attract solid particles on its outside surface. The particles were then dislodged into a 1.2-mL destination vial in a microbalance by spinning the pipette tip. Textures of solid that could be weighed included powder, crystalline, liquid, and semi-solid substances. The workstation can pick up submilligram quantities of sample (=0.3mg) from source vials containing as little as 1mg. The destination vials containing the samples were stored in a 96-well rack to enable subsequent automated liquid handling. Using bovine serum albumin as test solid, the coefficient of variation of the protein concentration for 48 samples is less than 6%. The workstation was used successfully to weigh out 48 different synthetic compounds. Time required for automated weighing was similar to manual weighing. The use of this workstation reduced 90% hands-on time and thus exposure to potentially toxic compounds. In addition, it minimized sample waste and reduced artifacts due to the poor solubility of compound in solvents. Moreover, it enabled compounds synthesized in milligram quantities to be weighed out and tested in biological assays.

Incorporation of 3T3-L1 cells to mimic bioaccumulation in a microscale cell culture analog device for toxicity studies.

Deficiencies in the early ADMET (absorption, distribution, metabolism, elimination, and toxicity) information on drug candidates extract a significant economic penalty on pharmaceutical firms. We have developed a microscale cell culture analog (microCCA) device that can potentially provide better, faster, and more efficient prediction of human and animal responses to a wide range of chemicals. The system described in this paper is a simple four-chamber microCCA ("lung"-"liver"-"fat"-"other tissue") designed on the basis of a physiologically based pharmacokinetics (PBPK) model of a rat. Cultures of L2, HepG2/C3A, and differentiated 3T3-L1 adipocytes were selected to mimic the key functions of the lung, liver, and fat compartments, respectively. Here, we have demonstrated the application of the microCCA system to study bioaccumulation, distribution, and toxicity of selected compounds. Results from the bioaccumulation study reveal that hydrophobic compounds such as fluoranthene preferentially accumulated in the fat chamber. Only a small amount of fluoranthene was observed in the liver and lung chambers. In addition, the presence of the differentiated 3T3-L1 adipocytes in the microCCA device significantly reduced naphthalene and naphthoquinone-induced glutathione (GSH) depletion. These findings suggest the potential utilization of the microCCA system to assess ADMET characteristics of the compound of interest prior to animal or human trials.

Development of a microscale cell culture analog to probe naphthalene toxicity.

Prediction of human response to drugs or chemicals is difficult as a result of the complexity of living organisms. We describe an in vitro model that can realistically and inexpensively study the adsorption, distribution, metabolism, elimination, and potential toxicity (ADMET) of chemicals. A microscale cell culture analog (microCCA) is a physical replica of the physiologically based pharmacokinetics (PBPK) model. Such a microfabricated device consists of a fluidic network of channels to mimic the circulatory system and chambers containing cultured mammalian cells representing key functions of animal "organ" systems. This paper describes the application of a two-cell system, four-chamber microCCA ("lung"-"liver"-"other tissue"-"fat") device for proof-of-concept study using naphthalene as a model toxicant. Naphthalene is converted into reactive metabolites (i.e., 1,2-naphthalenediol and 1,2-naphthoquinone) in the "liver" compartment, which then circulate to the "lung" depleting glutathione (GSH) in lung cells. Such microfabricated in vitro devices are potential human surrogates for testing chemicals and pharmaceutics for toxicity and efficacy.

Proyecto de una unidad modelo de farmacología clínica para evaluar medicamentos genéricos intercambiables / Project for a model clinical pharmacology unit to evoluate interchangeable generic drugs

La regulación que en materia de prescripción de medicamentos se ha establecido en la práctica pública y privada para su denominación genérica, hace necesario que los sectores involucrados: instituciones de salud, industria farmacéutica, universidad y comunidad usuaria, den cumplimiento a los procedimientos normativos en torno a este nuevo sistema. Con fundamento en lo anterior, se ha estructurado el Programa gubernamental sobre medicamentos genéricos intercambiables, con un enfoque dirigido a la validación de fármacos, sin soslayar el impacto económico y social que aportará en los aspectos de costo-beneficio, costo-eficacia y costo-calidad. Este proyecto innovador contempla además la elaboración de un catálogo de actualización dinámica de este tipo de medicamentos, la apertura de un mercado nacional y la creación y desarrollo de unidades de farmacología clínica para su evaluación. Ante la creciente demanda de servicios en el área de la farmacología clínica será prioritarios contar en México con suficientes centros de investigación de alta calidad en este campo. El Instituto Mexicano del Seguro Social presenta en este documento el proyecto de una unidad modelo de farmacología clínica, instrumentada con la infraestructura suficiente que le permita la evaluación de medicamentos genéricos que se acreditarán como intercambiables, ya que no existen procedentes en el país de una estructura operativa con estas características. Así mismo, estas unidades especializadas podrán realizar funciones de apoyo en el proceso de enseñanza, e investigación, fungir como centros de información de medicamentos, farmacovigilancia, farmacoepidemiología, farmacoeconomía y de referencia a nivel nacional e internacional

A fully automated light/dark apparatus useful for comparing anxiolytic agents.

The effects of known anxiolytic agents and putative anxiolytic agents were assessed in mice in a fully automated 2-compartment light/dark test. Significant increases in lit area activities (e.g., time spent in the lit area, locomotor activity, rearing behavior) were used as possible indicators of anxiolytic-like action. The measurement found most consistent and useful for assessing antianxiety-like activity was the time mice spent in the lit area. The benzodiazepine, diazepam; the 5-HT1A agent, ipsapirone; and the 5-HT3 receptor antagonist, ondansetron, produced significant anxiolytic-like activity between doses of 1.0 to 10.0 mg/kg, 17.8 to 31.6 mg/kg, and 0.0001 to 1.0 mg/kg respectively. The 5-HT1A receptor agonist, 8-OH DPAT, also exhibited anxiolytic-like action between doses of 0.0005 to 3.16 mg/kg. In contrast, the peripheral 5-HT3 receptor agonist, N-phenylbiguanide; the antidepressant, imipramine; the neuroleptic, chlorpromazine; and the CNS stimulant, S(+)-amphetamine, did not display antianxiety-like activity. The positive results obtained for the three types of compounds (benzodiazepine, 5-HT1A, and 5-HT3) indicate that this fully automated light/dark apparatus may be useful for identifying known and putative anxiolytic agents.

Cost considerations of ambulatory blood pressure monitoring.

Automatic blood pressure monitoring has proved valuable in testing new antihypertensive drugs, providing greater precision at less cost. Results obtained by this method are better correlated with target-organ damage (e.g. left ventricular hypertrophy) than conventional office blood pressure measurements. A number of clinical conditions are better evaluated with ambulatory blood pressure monitoring than with conventional measurements, e.g. office (white coat) hypertension, episodic hypertension and intermittent hypertensive symptoms. An economic model suggests that appropriately constrained ambulatory blood pressure monitoring can be highly cost-effective in patients who are initially diagnosed as mildly hypertensive on the basis of repeated casual blood pressure measurements. Normative data are needed for populations by age, race, gender, body habitus and conditions such as pregnancy.