The effect of small interfering RNA (siRNA) against the Bcl-2 gene on apoptosis and chemosensitivity in a canine mammary gland tumor cell line.

The aim of this study was to investigate whether downregulation of Bcl-2 expression by small interfering RNA (siRNA) against the canine Bcl-2 gene would enhance the apoptosis and sensitivity of a canine mammary gland tumor cell line (CF33) to doxorubicin. Transfections of CF33 with siRNA were performed using cationic liposomes. Sequence-specific downregulation of Bcl-2 expression was measured by semiquantitative RT-PCR and Western blot analysis. Total viable cells were determined by MTS assay and apoptotic cell rates were determined by the immunohistochemical analysis on ssDNA. Our data showed the siRNA downregulated Bcl-2 expression which increased apotosis and also increased the sensitivity of CF33 to doxorubicin. This study indicated that downregulation of Bcl-2 expression by siRNA would be useful as a new protocol to increase the effect of doxorubicin on treatment of canine mammary gland tumors, requiring a detailed evaluation of siRNA in vivo.

Apoptosis of canine mammary tumor cells induced by small interfering RNA (siRNA) against Bcl-xL gene.

The inhibition of Bcl-xL mRNA expression and the acceleration of apoptotic cell rates in canine mammary tumor cell line (CF33) by the small interfering RNA (siRNA) were analyzed. The level of Bcl-xL transcripts in CF33 was decreased when cultured with siRNA, suggesting that siRNA might inhibit the expression of Bcl-xL mRNA in the CF33. Apoptotic cell rates in CF33 cultured with siRNA in Oligofectamine medium, with double strand RNA in Oligofectamine medium, without siRNA in Oligofectamine medium and in DMEM alone were 60.9%, 30%, 28.7% and 11.6% at 48-hr incubation, respectively, when evaluated by TUNEL assay. From these results, it was suggested that canine Bcl-xL might be an anticancer target of canine tumors.

[The construction and evaluation of the preventing method for the input mischoice in a prescription order entry system--usefulness of a three-character input and a warning screen display system].

In the computerized prescription order entry system, it has been pointed out that a physician's input mischoice for medicine is one of the causes of medication errors. We therefore investigated the input mischoices by physicians at the time of writing prescriptions. Subsequently, the number of input characters in a prescription order was changed to three characters from two characters. Furthermore, 105 items of high-alert medications, which are likely to result in injury if errors occur, were established. A warning screen display system that requests reconfirmation of the effect, name, usage, and dosage of those medicines was also built. It was found that 70% of input mistakes were caused by choosing the medicine displayed immediately above or below the desired drug. By changing the number of input characters of a prescription order to three characters from two characters, the rate of specification of a trademark improved sharply from 36% to 85%. Consequently, the rate of choice of a drug with another trademark decreased significantly from 0.028% to 0.0047%. In 5% of cases when the warning screen was displayed for a high-alert medicine, the prescription was stopped, and 25% were changed to other medicines. The above results show that the system that requires the input of three or more characters for the physician order entry and displays a warning screen for high-alert medicines is useful in preventing mischoices at the time of prescription input.

[Information technology for ensuring drug safety].

In Japan, information technology (IT) in the medical field has prevailed as a means for handling claims for health insurance reimbursement. In contrast, IT is primarily used for electronic medical records in Western countries. Originally, preparation of health insurance claims was one of the outcomes of computerized medical information in Japan. As its protocols are already well established in Japan, information from the insurance claim system is hard to integrate into the Electronic Medical Chart system. To ensure drug safety, it is necessary to determine the number of users, and to accurately tabulate the incidence of adverse events. For this purpose, three kinds of information are required: prescription information, dispensing information, and drug administration information. Prescription information and dispensing information should be consistent with each other in content. Dispensing information is essential to identify the "substance" when adverse events occur. Drug administration information is the "true drug history". With these three kinds of information, it should be possible to enter drug safety data as evidence. To accurately capture these three kinds of information, it is necessary to utilize Standard Drug Code and Standard Usage Master, suggesting that it may be necessary to reconstruct the current system.

A proposal for a drug information database and text templates for generating package inserts.

To prevent prescription errors caused by information systems, a database to store complete and accurate drug information in a user-friendly format is needed. In previous studies, the primary method for obtaining data stored in a database is to extract drug information from package inserts by employing pattern matching or more sophisticated methods such as text mining. However, it is difficult to obtain a complete database because there is no strict rule concerning expressions used to describe drug information in package inserts. The authors' strategy was to first build a database and then automatically generate package inserts by embedding data in the database using templates. To create this database, the support of pharmaceutical companies to input accurate data is required. It is expected that this system will work, because these companies can earn merit for newly developed drugs to decrease the effort to create package inserts from scratch. This study designed the table schemata for the database and text templates to generate the package inserts. To handle the variety of drug-specific information in the package inserts, this information in drug composition descriptions was replaced with labels and the replacement descriptions utilizing cluster analysis were analyzed. To improve the method by which frequently repeated ingredient information and/or supplementary information are stored, the method was modified by introducing repeat tags in the templates to indicate repetition and improving the insertion of data into the database. The validity of this method was confirmed by inputting the drug information described in existing package inserts and checking that the method could regenerate the descriptions in the original package insert. In future research, the table schemata and text templates will be extended to regenerate other information in the package inserts.

Analysis on descriptions of precautionary statements in package inserts of medicines.

BACKGROUND: To prevent medical accidents, users must be informed of the cautions written in medical package inserts. To realize countermeasures by utilizing information systems, we must also implement a drug information database. However, this is not easy to develop, since the descriptions in package inserts are too complex and their information poorly structured. It is necessary to analyze package insert information and propose a data structure. METHODS: We analyzed the descriptions of 'precautions for application' in package inserts via text mining methods. In order to summarize statements, we applied dependency analysis to statements and visualized their relations between predicate words and other words. Furthermore, we extracted words representing timing to execute the order. RESULTS: WE FOUND THAT THERE ARE FOUR TYPES OF STATEMENTS: direct orders such as " " (use), causative orders such as " " (make someone use), direct interdictions such as " " (do not use), and causative interdictions such as " " (do not make user use). As for words representing timing, we extracted six groups: "at the time of delivery," "at the time of preparation," "in use," "after use," and "at the time of storage." From these results, we obtained points of consideration concerning the subjects of orders in the statements and timing of their execution. CONCLUSION: From the obtained knowledge, we can define the information structure used to describe the precautionary statement. It should contain information such as the actions described in the statement, the flag to express an order or interdiction, the subject to be ordered, and the timing.

Bactericidal effects of a high-power, red light-emitting diode on two periodontopathic bacteria in antimicrobial photodynamic therapy in vitro.

AIM: Light-emitting diodes have been investigated as new light activators for photodynamic therapy. We investigated the bactericidal effects of high-power, red light-emitting diodes on two periodontopathic bacteria in vitro. METHODS: A light-emitting diode (intensity: 1100 mW/cm(2) , peak wavelength: 650 nm) was used to irradiate a bacterial solution for either 10 or 20 s. Bacterial solutions (Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans) at a concentration of 2.5 × 10(6) c.f.u./mL were mixed with an equal volume of either methylene blue or toluidine blue O (0-20 µg/mL) and added to titer plate wells. The plate wells were irradiated with red light-emitting diode light from a distance of 22 or 40 mm. The contents were diluted, and 50 µL was smeared onto blood agar plates. After 1 week of culturing, bacterial c.f.u. were counted. RESULTS: The light-emitting diode energy density was estimated to be approximately 4 and 8 J/cm(2) after 10 and 20 s of irradiation, respectively. Red light-emitting diode irradiation for 10 s from a distance of 22 mm, combined with methylene blue at concentrations >10 µg/mL, completely killed Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. CONCLUSION: High-power, red light-emitting diode irradiation with a low concentration of dye showed effective bactericidal effects against two periodontopathic bacteria.

Voluntary medication error reporting program in a Japanese national university hospital.

BACKGROUND: In Japan, as in other countries, medical accidents arising from human error can seriously damage public confidence in medical services, as well as being intrinsically undesirable. OBJECTIVE: Errors voluntarily reported by the healthcare practitioners in our institution (Kanazawa University Hospital) were considered to assess the contributory factors by using the accumulated error database in the hospital information system. METHODS: Medical errors in our institution during the period from July 1, 2000, to June 30, 2002, were counted using the error reporting system database and were classified. RESULTS: The number of errors reported during the investigation period was 1378, of which 78% were reported by nursing staff. Medication errors involving administration of injectable or oral drugs to inpatients, dispensing, and prescription accounted for about 50% of that number. Among dispensing errors, 53% were detected by patients or their families and 36% by nurses. CONCLUSIONS: The best method of error prevention is to learn from previous errors. For this purpose, the error reporting program is effective. In patient safety management, it is important to take into account the potential risks of future errors, as well as to capture information about errors that have already happened. For safety management, adoption of appropriate information technology (e.g., implementation of a prescription order entry system) is effective in reducing medication errors. However, it is important to note that serious errors can also arise in computer-based systems.

Prediction and assessment of extrapyramidal side effects induced by risperidone based on dopamine D(2) receptor occupancy.

In the present study, we attempted to predict the risk of extrapyramidal side effects of a selective monoaminergic antagonist, risperidone, by analyzing the correlation between the dopamine D(2) receptor occupancy and the degree of extrapyramidal side effects of the drug. The occupancies of D(2) and 5-HT(2) receptors at various doses of risperidone were calculated by means of a receptor occupancy theory. The extrapyramidal side effects after administration of risperidone were attempted to predict by theoretical analysis of the correlation between the calculated occupancies and the evidence of extrapyramidal symptoms using a ternary complex model. The pharmacokinetic/pharmacodynamic analysis utilized the data concerning the pharmacokinetics of risperidone and 9-hydroxyrisperidone (active metabolite), their binding affinities with D(2) and 5-HT(2) receptors, and the clinical evidence of extrapyramidal symptoms (Extrapyramidal Symptom Rating Scale: ESRS), gathered from the literature. The mean occupancy of 5-HT(2) receptors after the administration of regular doses of risperidone was suggested to be more than 90%, whereas the mean occupancy of D(2) receptors varied between 50-80%, depending on the dose. The correlation between the occupancy of D(2) receptors and the extrapyramidal symptoms could be successfully analyzed with a ternary complex model, showing the predictability of the model for the extrapyramidal side effects of risperidone. Since the estimated risk of the extrapyramidal side effects varied with the dose, the present method of predicting the extrapyramidal side effects of risperidone may provide a basis for developing a rational dosing regimen for the drug.