Appendicular fracture repair in dogs using the locking compression plate system: 47 cases.

The locking compression plate (LCP) has combination screw holes, making it possible to use the implant in three different ways; as a pure internal fixator using locking head screws, as a conventional compression plate using compression screws, or as a hybrid of the two. The experience with the LCP system in veterinary fracture repair is limited. The objective of this study was to evaluate the outcome of appendicular fractures in dogs, which were repaired with the LCP system combined with less invasive surgical techniques. Medical records and radiographs from 47 dogs were studied retrospectively. Thirty-four percent of the fractures were simple, six percent wedge and 60% comminuted fractures of the humerus (11 %), radius and ulna (30 %), femur (34 %) and of the tibia and fibula (25 %). The fractures were treated using the LCP as an internal fixator; in some cases as a plate and rod construct. Forty-six of 47 fractures reached radiographic union. Mean healing time of the fractures was seven weeks (95% confidence interval from 5.8 to 8.3 weeks). There were statistically significant differences in healing time between juvenile (age under one year) and adults. Complications in the form of implant failures and infections were encountered in approximately 11% of the cases. All implant failures were due to surgical errors. The LCP system in combination with a less invasive surgical approach was found advantageous in comminuted fractures where the LCP was used as a bridging plate, in situations when exact plate contouring was difficult, and when other implants prevented the use of bi-cortical screws.

Luxation of the long digital extensor tendon as a complication to Tibial Plateau Levelling Osteotomy. A presentation of four cases.

Cranial cruciate ligament disease in dogs is frequently treated with Tibial Plateau Levelling Osteotomy (TPLO). Herein we describe four cases of dogs presenting with sudden lameness in the operated leg one to 12 months post TPLO surgery. On examination, all of the dogs had a luxation of the long digital extensor tendon (LDE) resulting from the TPLO surgery. All of the dogs underwent revision surgeries. The LDE tendon was either secured in its normal position or transected, and a tenodesis was performed. The dogs recovered well after surgery and lameness was resolved in all four cases.

Regression spline models and model calibration in the identification of protein storage conditions.

The study of protein-construct activity provides valuable insight in the early stages of drug discovery. Identification of optimal storage conditions that maintain activity of the constructs is an important issue. In the study reported herein, a space-filling design was used to assess the effects of eight design variables--buffer, pH, NaCl, protein concentration, reducing agent, detergent, MgCl2, and temperature--on protein activity. A regression spline analysis is presented, and settings of the explanatory factors resulting in the best predicted protein activity over the explored space are identified. The models are selected initially based on the Akaike information criterion and later assessed via root mean square error and cross-validation root mean square error. Detergent, buffer, temperature, and smooth functions of pH and protein concentration were found to have large effects on protein activity. The models were calibrated via calculation of the empirical distribution of the cross-validation root mean square error. In this framework several models provide a similar fit, and further experimentation is required for more definitive conclusions regarding protein storage conditions to be made. The cross-validation root mean square error calibration of models is recommended for applications involving comparisons of models with respect to their predictive ability.

Measurement of Clostridium perfringens beta-toxin production by surface plasmon resonance immunoassay.

A rapid and sensitive assay using surface plasmon resonance (SPR) immunoassay has been developed for the detection of Clostridium perfringens beta-toxin. The SPR immunoassay was conducted off-line by passing fermentation broth by a sensor chip coated with a monoclonal antibody specific for C. perfringens beta-toxin. Quantitation of toxin using SPR immunoassay was achieved by mass transport analysis; results were obtained within 20 min. The SPR immunoassay was compared with an ELISA and the traditional bioassay for C. perfringens beta-toxin. The SPR immunoassay and ELISA detected at least twofold differences in toxin levels at 95% confidence over a broad range of toxin concentrations. The traditional bioassay did not produce the resolution observed with the immunoassays. The SPR immunoassay allows for real-time monitoring of beta-toxin accumulation during production and permits the bioengineer to harvest C. perfringens fermentations when toxin is most concentrated. The SPR methodology may be applied to other fermentations to enhance and optimize toxin yields.

Quantitative reverse transcription strand displacement amplification: quantitation of nucleic acids using an isothermal amplification technique.

Recent advances in nucleic acid amplification techniques have allowed for quantitation of viral nucleic acid levels in clinical specimens. The most prevalent testing is carried out for HIV viral load. Strand displacement amplification (SDA) is an isothermal DNA amplification system utilizing a restriction enzyme and a DNA polymerase with strand displacement properties. SDA was adapted for quantitative RNA amplification (QRT-SDA) of an HIV gag sequence by including AMV reverse transcriptase, a quantitative control sequence, and 32P-labeled detector oligonucleotides for the HIV and the control sequences. We have also improved the amplification efficiency by including the single-strand binding protein from gene 32 of T4 bacteriophage (T4gp32) to enhance strand displacement replication. In a preliminary analytical demonstration of the technique, RT-SDA was quantitative to within twofold over a range of 500-500,000 transcripts that were generated from a plasmid bearing an HIV gag sequence. QRT-SDA potentially represents a convenient alternative for viral load testing in a clinical setting.

Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA.

A total of 294 clinical respiratory specimens, including 75 with culture-positive results, were tested for the presence of Mycobacterium tuberculosis by strand displacement amplification (SDA) of DNA. A region of the IS6110 insertion element and an internal control sequence were amplified and then detected by a chemiluminescence assay. Receiver operator-characteristic curves were used to evaluate three methods for declaring specimens positive for M. tuberculosis. By the preferred method, SDA chemiluminescence results were converted to theoretical numbers of M. tuberculosis organisms. A positive threshold (PT) value, above which 95% of the SDA results were judged to be M. tuberculosis positive (sensitivity = 95%), was found to be 2.4 M. tuberculosis organisms per SDA reaction. The analogous PT value for 95% sensitivity on smear-positive specimens was 3.6 M. tuberculosis organisms per reaction. The PT of 2.4 M. tuberculosis organisms per reaction detected 100% of culture-positive, smear-positive specimens (sensitivity = 100%), while 95% sensitivity was achieved with a PT of 15.5 M. tuberculosis organisms per reaction. Specificities, which were calculated with respect to culture- and smear-negative specimens, ranged from 96% at a PT of 15.5 M. tuberculosis organisms to 84% at a PT of 2.4 M. tuberculosis organisms per reaction. The M. tuberculosis-negative specimens were also segregated according to whether the patients received antituberculosis chemotherapy. SDA specificity ranged from 90% (PT = 2.4 M. tuberculosis organisms) to 98% (PT = 15.5 M. tuberculosis organisms) for the M. tuberculosis-negative specimens from patients who had not received chemotherapy. SDA specificity in the M. tuberculosis-negative specimens from patients who received chemotherapy was lower (85 to 94%). This study represents the first large-scale demonstration of M. tuberculosis detection in clinical sputum specimens by isothermal DNA amplification with SDA.

Virtual kinetics: using statistical experimental design for rapid analysis of enzyme inhibitor mechanisms.

Modern automated drug-screening can generate hundreds of inhibitor leads from diverse chemical sources in a short period of time. Traditional methods of inhibitor analysis are resource intensive and limit the number of inhibitors that can be analyzed for their mechanism of inhibition. This paper presents methods we have developed for rapid estimation of both potency and mechanism of potential inhibitor leads for a biochemically complex screening target (protein kinase C) using commercially available computer programs for statistical experimental design. Our findings indicate that, with careful choice of factor levels, statistical experimental design clearly identifies the various interactions of the assay components with inhibitors. Suitably plotted, the data can be used to examine the competitive nature of the inhibitor and can provide estimates of IC50 and Michaelis constants useful for planning further kinetic work. The techniques used are amenable to automation and should be useful for identifying inhibitors that may have only marginal potency, but exhibit desirable mechanistic profiles suitable for structural analoging efforts.

Chemiluminescent detection of strand displacement amplified DNA from species comprising the Mycobacterium tuberculosis complex.

Strand displacement amplification, a new isothermal in vitro DNA amplification technique, was used to amplify target DNA contained within the IS6110 insertion element of the species within the Mycobacterium complex (Mycobacterium tuberculosis, M. bovis, M. bovis-BCG, M. africanum and M. microti). The target nucleic acid sequence is present in approximately ten, two, one, five and five copies in M. tuberculosis, M. bovis, M. bovis-BCG, M. africanum and M. microti, respectively. Amplified products were detected using a non-isotopic microtitre plate assay employing a biotinylated oligodeoxynucleotide probe and an alkaline phosphatase conjugated oligodeoxynucleotide probe. Lumiphos 530 was the chemiluminescent substrate for alkaline phosphatase. The combination of the strand displacement amplification method with this sensitive and rapid (less than 2 h) detection system resulted in the specific detection of as few as 1-25 initial IS6110 targets in the five Mycobacterium complex species based on signal/noise criteria. Negative results were obtained with eight other Mycobacterium species as well as with 32 non-Mycobacterium species.

[Silicone breast implants and collagen diseases]. / Silikon mammaproteser og kollagene sykdommer.

Connective tissue disease has been reported to occur following implantation of silicone gel-filled prostheses to augment the breast. In this case report and review of the literature, two patients are described in whom connective tissue disease developed within one and three years respectively after cosmetic surgery. One developed dermatomyositis with lung fibrosis, and the other developed cutaneous lupus and a Sjögren-like syndrome. Both suffered serious complications due to vasculitis. Although evidence of a causal relationship between the implantation and the development of connective tissue disease is circumstantial, removal of the silicone prosthesis has been reported to result in subsequent remission.

Analysis of the variance components in a pharmaceutical aerosol product: lodoxamide tromethamine.

The contributions of several components to the variance in lodoxamide delivery from lodoxamide tromethamine metered-dose aerosol containers have been estimated. Two aerosol lots, manufactured with mean diameters of 2.3 and 7.2 micron, exhibited approximately equal variances. The variance was apportioned to the following components: container-to-container, 27%; mouthpiece-to-mouthpiece 18%; valve-to-valve, 11%; assay, 6%. The largest single contribution to the variance (38%) is attributed to unassignable variations which include within-container variations in the dose; quality improvement efforts should concentrate on this area. Little effort should be expended to minimize the assay, valve delivery, or mouthpiece variation as their contribution to lodoxamide dose variation is small. Likewise, the bulk drug particle size did not contribute appreciably to within-lot dose variation.

Interns' evaluation of their preparation for general practice: a comparison between the University of Tromsø and the University of Bergen.

A total of 168 interns who have graduated from the Medical Schools of Bergen and Tromsø were asked about various aspects of the medical curriculum. In Bergen the curriculum has a traditional structure with a pre-clinical and a clinical part, but in Tromsø the pre-clinical and clinical subjects are integrated. In addition, the students in Tromsø spend long periods in municipal hospitals and in the primary health care service. We were interested in how the interns from the two universities evaluated their respective curricula and how prepared they felt for their current work. There was a response rate of 86% to the questionnaire. The results showed that the interns from Tromsø are more satisfied with their education and feel more confident in their practical skills than the interns from Bergen. They are also more motivated for future work in general practice. In our opinion the main reason for these results is the difference in curricula in the two medical schools. Other possible reasons are also discussed.