Utilização de medicamentos e risco de interações medicamentosas em idosos atendidos pelo Programa de Atenção ao Idoso da Unijuí / Use of medications and risk of drug interactions in the elderly population at the Elderly Care Program in Unijuí

Idosos utilizam elevado número de medicamentos, apresentando alto potencial para desenvolver interações medicamentosas. Objetivou-se verificar o perfil de utilização de medicamentos e conhecer possíveis interações medicamentosas em idosos acompanhados pelo Programa de Atenção ao Idoso (PAI) da Unijuí. Os dados foram coletados em julho de 2009. Classificaramse os medicamentos no sistema Anatomical Therapeutic Chemical e possibilidades de interações segundo Drug Interaction Facts. De março/2008 até julho/2009 o PAI atendeu 31 idosos, estando 16 em acompanhamento, os quais apresentaram média de 78,4 ± 6,8 anos e utilizaram 83 medicamentos, com média de 5,2 ± 3,7/ idoso. Os fármacos mais prevalentes foram os que atuam no aparelho cardiovascular, no sistema nervoso e para o trato alimentar e metabolismo. Verificaram-se 36 possíveis interações entre 9 pacientes, com média de 4/idoso. O fármaco mais envolvido foi digoxina. Quanto a classificação, 4 apresentaram nível de significância 1, dez nível 2, cinco nível 3, sete nível 4 e dez nível 5. O número de possíveis interações verificadas foi expressivo, entretanto nem todos os idosos apresentarão reações relacionadas às interações, mas apresentaram o risco. Portanto, é importante que prescritores conheçam a influência de um fármaco sobre outro. Seria necessário treinamento intensivo para intervenção farmacêutica, sendo o relacionamento com o médico indispensável.

Elderly people take a large number of medicines, entailing a high risk of developing drug interactions. The aim was to study the profile of medicines prescribed and the possible drug interactions in the public Elderly Care Program (PAI) run at Unijuí, a university in south Brazil. The data were collected in July 2009. The Anatomical Therapeutic Chemical (ATC) Classification System was used to classify medicines, while the potential interactions were classified with the aid of Drug Interaction Facts. Between March 2008 and July 2008, the PAI attended 31 elderly patients, of whom 16 were monitored, with a mean age of 78.4 ± 6.8 years. This group received 83 medicines, making an average of 5.2 ± 3.7 medicines/patient. The most prevalent drugs were those acting on the cardiovascular system, nervous system, digestive tract and metabolism. There were 36 possible interactions in 9 patients, an average of 4 interactions per patient. The drug most frequently involved was digoxin. The interactions were classified as follows: 4 at level 1, 10 at level 2, 5 at level 3, 7 at level 4 and 10 at level 5. This number of possible interactions is considerable; although not all elderly patients show the effects of the drug interactions, they do run the risk. Therefore, it is highly important that prescribers are familiar with the drug interactions. For the pharmacist to help the patient, intensive training for pharmaceutical intervention and a good relationship with the doctor are indispensable.

The Heidelberg Ion Therapy Center.

The ion beam therapy facility presently under construction at the Department of Clinical Radiology, University of Heidelberg, Germany, will be the first dedicated and hospital-based irradiation facility for protons and heavier ions in Europe. A capacity of more than 1000 patient treatments per year is planned. The facility comprises two horizontally-fixed beamlines for patient treatments plus a fixed-beam experimental area. In addition, the world-wide first scanning ion gantry is under construction. The facility fully relies on an active beam delivery method, the intensity-controlled rasterscan technique. The availability of different ion species ranging from protons to oxygen under identical conditions optimally supports clinical trials aiming to clarify the question of which particle species is best suited for the individual indications. A linac-synchrotron combination will deliver libraries of energy-, focus- and intensity-variable pencil-beams for each ion species to the dose-delivering scanning systems at each treatment station. The available energies correspond to water-equivalent ranges from 2 cm to 30 cm. The intensity-controlled rasterscan technique allows for the administration of inversely planned and biologically optimized dose distributions having utmost precision. The facility will be equipped with state-of-the-art imaging modalities as well as an in-situ Positron-Emission-Tomography (PET). The commissioning of the different sections is scheduled for 2006. The pre-clinical operation will start early in 2007 followed by the routine patient treatment.

Adjustment of transfer tools for the production of micro- and macroarrays.

A transfer tool adjustment procedure for the generation of micro- and macroarrays is described. It is based on control spotting of solutions containing radioactive or fluorescent labels and the quantification of each obtained spot by standard image-analyzing software. This method provides a simple, rapid, and efficient way to control the quality and liquid delivery properties of spotting transfer tools.

New tools for oligonucleotide fingerprinting.

Oligonucleotide fingerprinting is an attractive, high-throughput complement to tag sequencing methods to determine the spectrum and abundance of genes in cDNA libraries. This method currently relies on the sequential hybridizations of short, radioactively labeled DNA oligonucleotides to clone arrays. Here, we describe a new environment that substantially improves this technology. Fluorescently labeled peptide nucleic acid (PNA) oligonucleotides are used as hybridization probes. Hybridization results are recorded with a large-field, high-resolution laser scanner developed for this purpose. Automated image analysis allows easy handling of large numbers of hybridization images. Signal interference effects, which limit the gridding density in the radioactive approach, are strongly reduced. The sensitivity of the fluorescence detection demonstrated permits the convenient use of nylon membranes. Hybridization data quality is improved, and its generation is substantially accelerated, simplified, and less expensive.

Large-gel two-dimensional electrophoresis-matrix assisted laser desorption/ionization-time of flight-mass spectrometry: an analytical challenge for studying complex protein mixtures.

The large-gel two-dimensional electrophoresis (2-DE) technique, developed by Klose and co-workers over the past 25 years, provides the resolving power necessary to separate crude proteome extracts of higher eukaryotes. Matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) provides the sample throughput necessary to identify thousands of different protein species in an adequate time period. Spot excision, in situ proteolysis, and extraction of the cleavage products from the gel matrix, peptide purification and concentration as well as the mass spectrometric sample preparation are the crucial steps that interface the two analytical techniques. Today, these routines and not the mass spectrometric instrumentation determine how many protein digests can be analyzed per day per instrument. The present paper focuses on this analytical interface and reports on an integrated protocol and technology developed in our laboratory. Automated identification of proteins in sequence databases by mass spectrometric peptide mapping requires a powerful search engine that makes full use of the information contained in the experimental data, and scores the search results accordingly. This challenge is heading a second part of the paper.

The novel transcription factor e(y)2 interacts with TAF(II)40 and potentiates transcription activation on chromatin templates.

Weak hypomorph mutations in the enhancer of yellow genes, e(y)1 and e(y)2, of Drosophila melanogaster were discovered during the search for genes involved in the organization of interaction between enhancers and promoters. Previously, the e(y)1 gene was cloned and found to encode TAF(II)40 protein. Here we cloned the e(y)2 gene and demonstrated that it encoded a new ubiquitous evolutionarily conserved transcription factor. The e(y)2 gene is located at 10C3 (36.67) region and is expressed at all stages of Drosophila development. It encodes a 101-amino-acid protein, e(y)2. Vertebrates, insects, protozoa, and plants have proteins which demonstrate a high degree of homology to e(y)2. The e(y)2 protein is localized exclusively to the nuclei and is associated with numerous sites along the entire length of the salivary gland polytene chromosomes. Both genetic and biochemical experiments demonstrate an interaction between e(y)2 and TAF(II)40, while immunoprecipitation studies demonstrate that the major complex, including both proteins, appears to be distinct from TFIID. Furthermore, we provide genetic evidence suggesting that the carboxy terminus of dTAF(II)40 is important for mediating this interaction. Finally, using an in vitro transcription system, we demonstrate that recombinant e(y)2 is able to enhance transactivation by GAL4-VP16 on chromatin but not on naked DNA templates, suggesting that this novel protein is involved in the regulation of transcription.

Development of a technology for automation and miniaturization of protein crystallization.

The usage of standard 96 well microplates for the screening of crystallization conditions of recombinant proteins offers several advantages when compared to commonly used crystallization plate formats. The adoption of robotic technology for plate and glass slide preparation within a "hanging drop" vapour diffusion crystallization experiment enables to work with an increased throughput at reduced costs. In addition to commercial pipetting devices with a 96-channel aspirator/dispenser, solenoid ink-jet technology was applied to form 250 nl droplets with a diameter of approximately 1 mm. This allows miniaturization of crystallization screening set-ups with an estimated ten-fold cost reduction when compared to commonly used 24 well plates.

Induction of putative tumor-suppressing genes in Rat-1 fibroblasts by oncogenic Raf-1 as evidenced by robot-assisted complex hybridization.

The growth factor receptor-dependent protein kinase Raf-1 is activated by GTP-bound Ras, thereby activating the mitogen-activated protein kinase pathway. To study the role of Raf in transformation we transduced Rat-1 cells with a tetracycline-regulatable retroviral vector encoding the constitutively active oncogenic C-terminal fragment of the human Raf-1 protein. Using subtractive hybridization of mRNAs from induced and noninduced cells and robot-assisted screening by complex hybridization, Raf-induced genes with various different characteristics of induction were investigated. Among the strongly induced genes were those involved in carcinogenesis such as metalloproteinases 3, 10 and 13, cathepsin L, ornithine decarboxylase, and putative tumor-suppressing genes such as monocyte chemoattracting protein 1, interferon-induced protein 10, a recently identified 2'-5' oligoadenylate synthetase-like protein, and plasminogen activator inhibitor type 2. Other components of the plasminogen activator system were not induced. Plasminogen activator inhibitor type 2 is a down-regulator of the proteolytic cascade consisting of various metalloproteinases, some of which are induced by a carboxy-terminal Raf mutant (RafCT). In conclusion, RafCT induces factors which act in a conflicting manner in respect of carcinogenesis, especially within the proteolytic system of the extracellular matrix.

Tissue gene expression analysis using arrayed normalized cDNA libraries.

We have used oligonucleotide-fingerprinting data on 60,000 cDNA clones from two different mouse embryonic stages to establish a normalized cDNA clone set. The normalized set of 5,376 clones represents different clusters and therefore, in almost all cases, different genes. The inserts of the cDNA clones were amplified by PCR and spotted on glass slides. The resulting arrays were hybridized with mRNA probes prepared from six different adult mouse tissues. Expression profiles were analyzed by hierarchical clustering techniques. We have chosen radioactive detection because it combines robustness with sensitivity and allows the comparison of multiple normalized experiments. Sensitive detection combined with highly effective clustering algorithms allowed the identification of tissue-specific expression profiles and the detection of genes specifically expressed in the tissues investigated. The obtained results are publicly available (http://www.rzpd.de) and can be used by other researchers as a digital expression reference.

Prestructured MALDI-MS sample supports.

Prestructured MALDI-MS sample supports have been developed that simplify high-throughput analysis of biomolecules and improve the detection sensitivity. The mass spectrometric sample support is coated with a thin layer of hydrophobic Teflon that carries an array of 200-microm gold spots, which provide hydrophilic sample anchors. Each transferred sample droplet contacts one anchor, on top of which, after solvent evaporation, the sample is exclusively deposited due to the strongly water repellent nature of the Teflon surface. The initial matrix concentration is kept low, enabling sample up-concentration by more than 2 orders of magnitudes before crystallization commences. As a result, the detection sensitivity is improved as documented by mass spectra recorded from 100 amol of various peptides, 1 fmol of a DNA 20 mer, and 5 fmol of a 130 bp PCR product. Size and spacing of the hydrophilic anchors are optimized for MALDI-MS performance (sample spot size approximately = laser irradiation spot size), for short analysis times (predetermined sample coordinates), and for high throughput sample preparation (sample anchor array according to the 1536 microtiter plate format).

Normalization strategies for cDNA microarrays.

Multiple Arabidopsis thaliana clones from an experimental series of cDNA microarrays are evaluated in order to identify essential sources of noise in the spotting and hybridization process. Theoretical and experimental strategies for an improved quantitative evaluation of cDNA microarrays are proposed and tested on a series of differently diluted control clones. Several sources of noise are identified from the data. Systematic and stochastic fluctuations in the spotting process are reduced by control spots and statistical techniques. The reliability of slide to slide comparison is critically assessed within the statistical framework of pattern matching and classification.

A fully automated multicapillary electrophoresis device for DNA analysis.

We describe the construction and performance of a fully automated multicapillary electrophoresis system for the analysis of fluorescently labeled biomolecules. A special detection system allows the simultaneous spectral analysis of all 96 capillaries. The main features are true parallel detection without any moving parts, high robustness, and full compatibility to existing protocols. The device can process up to 40 microtiter plates (96 and 384 well) without human interference, which means up to 15,000 samples before it has to be reloaded.

The GSI Cancer Therapy Project.

At the Heavy Ion Research Institute GSI in Darmstadt an experimental cancer treatment program with a five years duration has been developed. A new method for cancer treatment with ions is applied, using rasterscan method in addition to an active pulse to pulse variation of ion beam properties, including the energy, intensity and focusing. An overview of this Cancer Therapy Project is presented, that covers both accelerator aspects to provide the required beam variations within a short time and the installations at the treatment place for rasterscan control. In addition to a description of the technical design (control-hard- and software) experimental results will be shown, containing the achieved beam properties and measurements of rasterscan performance.

Hamstring release and posterior capsulotomy for fixed knee flexion contracture in haemophiliacs.

Between 1977 and 1981 hamstring release (n = 27) and posterior capsulotomy (n = 22) were performed on 27 haemophilic patients. The follow-up of 19 patients is documented for at least 5 years after the operation. The mean age was 30 years (15-40 years), the mean follow-up period was 12.5 years (5-20 years). Postoperatively 16 of 19 knee joints achieved immediate full extension, three patients showed 10 degrees extension deficiency. In the long run the original good result could be demonstrated in just 11 patients. Using the Orthopedic Advisory Committee of the World Federation of Hemophilia (OAC) Scores there were three good, 12 satisfactory and four poor results. In minor forms of arthropathy a hamstring release with posterior capsulotomy is the only recommended operation. In severe forms we would actually recommend a combination with other procedures such as arthroscopic synovectomy or joint replacement.

Protein microarrays for gene expression and antibody screening.

Proteins translate genomic sequence information into function, enabling biological processes. As a complementary approach to gene expression profiling on cDNA microarrays, we have developed a technique for high-throughput gene expression and antibody screening on chip-size protein microarrays. Using a picking/spotting robot equipped with a new transfer stamp, protein solutions were gridded onto polyvinylidene difluoride filters at high density. Specific purified protein was detected on the filters with high sensitivity (250 amol or 10 pg of a test protein). On a microarray made from bacterial lysates of 92 human cDNA clones expressed in a microtiter plate, putative protein expressors could be reliably identified. The rate of false-positive clones, expressing proteins in incorrect reading frames, was low. Product specificity of selected clones was confirmed on identical microarrays using monoclonal antibodies. Cross-reactivities of some antibodies with unrelated proteins imply the use of protein microarrays for antibody specificity screening against whole libraries of proteins. Because this application would not be restricted to antigen-antibody systems, protein microarrays should provide a general resource for high-throughput screens of gene expression and receptor-ligand interactions.

SH3GL3 associates with the Huntingtin exon 1 protein and promotes the formation of polygln-containing protein aggregates.

The mechanism by which aggregated polygins cause the selective neurodegeneration in Huntington's disease (HD) is unknown. Here, we show that the SH3GL3 protein, which is preferentially expressed in brain and testis, selectively interacts with the HD exon 1 protein (HDex1p) containing a glutamine repeat in the pathological range and promotes the formation of insoluble polyglutamine-containing aggregates in vivo. The C-terminal SH3 domain in SH3GL3 and the proline-rich region in HDex1p are essential for the interaction. Coimmunoprecipitations and immunofluorescence studies revealed that SH3GL3 and HDex1p colocalize in transfected COS cells. Additionally, an anti-SH3GL3 antibody was also able to coimmunoprecipitate the full-length huntingtin from an HD human brain extract. The characteristics of the interaction between SH3GL3 and huntingtin and the colocalization of the two proteins suggest that SH3GL3 could be involved in the selective neuronal cell death in HD.

Control of the synovium in haemophilia.

Adequate factor substitution is a prerequisite for successful conservative or operative treatment of haemophilic synovitis. Subsequent prophylactic factor substitution can prevent further joint problems in the future. Orthopaedic treatment regimes are dependent on the classification of the synovitis and the stage of the arthropathy. Synovitis can be classified into acute and chronic forms. The arthropathy is classified according to Arnold and Hilgartner. When the synovitis has already become chronic, early arthroscopic synovectomy is recommended in order to prevent the otherwise inexorable progress of the arthropathy. Failure to treat in the early phase of the pathology causes problems in the correction of further stages. The progression of the joint disease can be shown both clinically and radiologically as the arthropathy develops, and influences the treatment.

Arthroscopy for chronic hemophilic synovitis of the knee.

Between 1988 and 1995, 32 knee joints (29 patients) with hemophilic arthropathy underwent arthroscopy. The spectrum of procedures ranged from resection of fibrous plicae to synovectomy. Technical difficulties appeared in cases of scarred fixed patella, pronounced posterior tibia subluxation, and severe fibrous ankylosis. All operations done between 1988 and 1991 (23 operations; 21 patients) were reviewed retrospectively. The mean age of the patients in this series was 30 years and the mean followup was 5 years. On subjective evaluation, 13 operations achieved a definite improvement, five showed slight improvement, and two had no improvement (three operations were excluded). Arthroscopic surgery, as a relatively low risk technique, combined with early functional rehabilitation, can be used to achieve satisfactory results in patients with hemophilic arthropathy.