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PubMed1998

Bone assessment after total knee arthroplasty by dual-energy X-ray absorptiometry: analysis protocol and reproducibility.

Bone quality is important for the success of joint prostheses implantation, and the assessment of bone density after total knee arthroplasty by means of dual-energy X-ray absorptiometry may be useful for monitoring implant stability. The aim of this study is to suggest a validated analysis protocol for the assessment of bone status after total knee arthroplasty. A dedicated densitometric analysis protocol of five regions of interest was designed, and 10 subjects who had received an uncemented knee prosthesis (8 females and 2 males, aged 55-74 years) underwent three consecutive scans in posteroanterior and lateral projections, with repositioning after each scan to test the suitability and reproducibility of the protocol. The reproducibility of the measurement of bone mineral content and density in the femoral and tibial regions ranged, respectively, from 2.1% to 4.1%, from 0.9% to 2.6% for the posteroanterior scans, and from 2.7% to 5.6% and from 2.3% to 4.7% for the lateral scans, depending on the considered region. Our results confirm that the suggested protocol allows precise assessment of bone mineral content and density, and that dual-energy X-ray absorptiometry is reliable for the evaluation of bone mass around prosthetic implants.

Absorptiometry, Photon↗

PubMed2003

Assessment of image quality of a standard and two dose-reducing protocols in paediatric pelvic CT.

BACKGROUND: Concerns exist regarding the effect of radiation dose from paediatric pelvic CT scans and the potential later risk of radiation-induced neoplasm and teratogenic outcomes in these patients. OBJECTIVE: To assess the diagnostic quality of CT images of the paediatric pelvis using either reduced mAs or increased pitch compared with standard settings. MATERIALS AND METHODS: A prospective study of pelvic CT scans of 105 paediatric patients was performed using one of three protocols: (1) 31 at a standard protocol of 200 mA with rotation time of 0.75 s at 120 kVp and a pitch factor approximating 1.4; (2) 31 at increased pitch factor approaching 2 and 200 mA; and (3) 43 at a reduced setting of 100 mA and a pitch factor of 1.4. All other settings remained the same in all three groups. Image quality was assessed by radiologists blinded to the protocol used in each scan. RESULTS: No significant difference was found between the quality of images acquired at standard settings and those acquired at half the standard mAs. The use of increased pitch factor resulted in a higher proportion of poor images. CONCLUSIONS: Images acquired at 120 kVp using 75 mAs are equivalent in diagnostic quality to those acquired at 150 mAs. Reduced settings can provide useful imaging of the paediatric pelvis and should be considered as a standard protocol in these situations.

Age Factors↗

PubMed2003

Investigating a new cryopreservation protocol for yams ( Dioscorea spp.).

Experiments were initiated to develop protocols for four species of Dioscorea, namely D. bulbifera L., D. oppositifolia L., D. alata L. and D. cayenensis Lam., and to assess the effects of various factors on the survival and regrowth of explants after rewarming. The treatments consisted of three different methods-vitrification, droplet and a modified droplet method-as well as the following variables-cryoprotective solution, sucrose concentration in the preadaptation medium and cold acclimation. While most of the protocols resulted in low to zero survival and regrowth rates, maximum survival rates of 100% and 75% were obtained for D. oppositifolia L. and D. cayenensis Lam., respectively, using two different protocols of the modified droplet method. A higher average survival rate was obtained using the droplet and modified droplet techniques than for the original vitrification methods. When the average of all the experiments was taken, more than one-half (52%) of the surviving explants developed further within 1 month with the modified droplet method compared to zero regrowth with the two other methods. The optimum protocol for cryopreservation is specific for each genotype.

Cryopreservation↗

PubMed2005

Successful development of a shed-microspore culture protocol for doubled haploid production in Indonesian hot pepper (Capsicum annuum L.).

Various systems of anther and microspore cultures were studied to establish an efficient doubled haploid production method for Indonesian hot pepper (Capsicum annuum L.). A shed-microspore culture protocol was developed which outperformed all the previously reported methods of haploid production in pepper. The critical factors of the protocol are: selection of flower buds with more than 50% late unicellular microspores, a 1 day 4 degrees C pretreatment of the buds, followed by culture of the anthers in double-layer medium system for 1 week at 9 degrees C and thereafter at 28 degrees C in continuous darkness. The medium contained Nitsch components and 2% maltose, with 1% activated charcoal in the solid under layer and 2.5 muM zeatin and 5 muM indole-3-acetic acid in the liquid upper layer. All the ten genotypes of hot pepper tested, responded to this protocol. The best genotypes produced four to seven plants per original flower bud. This protocol can be used as a potential tool for producing doubled haploid plants for hot pepper breeding.

Capsicum↗

PubMed2006

Improved methods in Agrobacterium-mediated transformation of almond using positive (mannose/pmi) or negative (kanamycin resistance) selection-based protocols.

A protocol for Agrobacterium-mediated transformation with either kanamycin or mannose selection was developed for leaf explants of the cultivar Prunus dulcis cv. Ne Plus Ultra. Regenerating shoots were selected on medium containing 15 muM kanamycin (negative selection), while in the positive selection strategy, shoots were selected on 2.5 g/l mannose supplemented with 15 g/l sucrose. Transformation efficiencies based on PCR analysis of individual putative transformed shoots from independent lines relative to the initial numbers of leaf explants tested were 5.6% for kanamycin/nptII and 6.8% for mannose/pmi selection, respectively. Southern blot analysis on six randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene in each, and five randomly chosen lines identified to contain the pmi transgene by PCR showed positive hybridisation to a pmi DNA probe. The positive (mannose/pmi) and the negative (kanamycin) selection protocols used in this study have greatly improved transformation efficiency in almond, which were confirmed with PCR and Southern blot. This study also demonstrates that in almond the mannose/pmi selection protocol is appropriate and can result in higher transformation efficiencies over that of kanamycin/nptII selection protocols.

Agrobacterium tumefaciens↗

PubMed2003

Individually weight-adapted examination protocol in retrospectively ECG-gated MSCT of the heart.

The standard protocol in multislice spiral CT (MSCT) angiography for coronary arteries with fixed tube current-time settings leads to an overexposure and thus to an unnecessary high radiation dose in patients with lower weight when compared to heavier patients. The purpose of this study was to estimate the effect of reducing the radiation dose by adapting the tube current-time settings individually. Fifty patients underwent retrospectively ECG-gated MSCT of the heart. In 25 patients (group A1) a standard protocol with constant tube current-time settings was used (4 x 1-mm collimation, 120 kV, 400 mAs(eff)). Subsequently, artificial image noise was added to the data of these patients simulating a directive for weight-adapted tube current-time settings (group A2). In the other 25 patients (group B) an alternative protocol with individually weight-adapted tube current-time settings was applied. The data of all groups were evaluated by a regression analysis. The image quality was assessed objectively by measuring the CT attenuation in standardised regions of interest and subjectively by three radiologists using a five-point scoring system in a consensus reading. Applying the weight-adapted tube current-time settings the effective radiation dose was reduced by 17.9% for men and 26.3% for women. The standard protocol leads to an overexposure in light patients as seen in the plot of noise vs weight (slope 0.16+/-0.07 HU/kg). By applying the weight-adapted tube current-time settings a weight-independent, constant image noise is achieved (slope 0.04+/-0.1 HU/kg). Diagnostic image quality was preserved in all patients. Individually weight-adapted tube current-time settings allow for a substantial dose reduction when performing retrospectively ECG-gated MSCT angiography for coronary arteries without impairment of diagnostic image quality.

Aged↗

PubMed2004

The effect of varying low-dose protocols on perceived image quality in multidetector CT in a rabbit model of acute appendicitis.

The aim of this study was to evaluate the value of multidetector CT (MD-CT) with low-dose protocols in the diagnosis of acute appendicitis in a rabbit model. Ten New Zealand white rabbits underwent ligation of the appendix. MD-CT (Siemens Somatom Plus 4 Volume Zoom) was performed using three low-dose protocols: 80 kV/70 mAs; 80 kV/50 mAs; and 80 kV/20 mAs. Axial and coronal reformations were performed. Three experienced radiologists evaluated the images using a four-grade score: (1) excellent; (2) good; (3) moderate; and (4) not sufficient. Diagnosis was confirmed histopathologically. The overall score for the 80 kV/70 mAs was 1, for the 80 kV/50 mAs protocol 1.3, and the 80 kV/20 mAs protocol reached an overall score of 2. The coronal reformations reached with 1.1 (80 kV/70 mAs), 1.2 (80 kV/50 mAs), and 1.4 (80 kV/20 mAs) had a slightly better median score than the axial reconstructions. In cases of inconclusive ultrasound and clinical presentation, the use of MD-CT can be considered when evaluating children for suspected appendicitis.

Acute Disease↗

PubMed2006

A protocol for high-throughput phenotyping, suitable for quantitative trait analysis in mice.

Whole-genome genetic association studies in outbred mouse populations represent a novel approach to identifying the molecular basis of naturally occurring genetic variants, the major source of quantitative variation between inbred strains of mice. Measuring multiple phenotypes in parallel on each mouse would make the approach cost effective, but protocols for phenotyping on a large enough scale have not been developed. In this article we describe the development and deployment of a protocol to collect measures on three models of human disease (anxiety, type II diabetes, and asthma) as well as measures of mouse blood biochemistry, immunology, and hematology. We report that the protocol delivers highly significant differences among the eight inbred strains (A/J, AKR/J, BALBc/J, CBA/J, C3H/HeJ, C57BL/6 J, DBA/2 J, and LP/J), the progenitors of a genetically heterogeneous stock (HS) of mice. We report the successful collection of multiple phenotypes from 2000 outbred HS animals. The phenotypes measured in the protocol form the basis of a large-scale investigation into the genetic basis of complex traits in mice designed to examine interactions between genes and between genes and environment, as well as the main effects of genetic variants on phenotypes.

Animals↗

PubMed2005

Implementation of the modified-SHIRPA protocol for screening of dominant phenotypes in a large-scale ENU mutagenesis program.

SHIRPA is a three-stage protocol for the comprehensive assessment of primarily mouse behavior. The first stage consists of high-throughput phenotyping of 33 behavioral observations and 7 metabolic or disease observations. We modified this part of the protocol by integrating new morphologic observations into the initial phenotype assay of behavior and dysmorphology. Behavioral observations assessed by this protocol, now referred to as the "modified-SHIRPA," are compatible with the original "SHIRPA" protocol. Using modified-SHIRPA, we screened dominant phenotypes of more than 10,000 G(1) progeny generated by crossing DBA/2J females with ENU-treated C57BL/6J males. To date, we have obtained 136 hereditary-confirmed mutants that exhibit behavioral and morphologic defects. Some independent mutant lines exhibited similar phenotypes, suggesting that they may represent alleles of the same gene or mutations in the same genetic pathway. They could hold great potential for the unraveling of the molecular mechanisms of certain phenotypes.

Animals↗

PubMed2001

Perioperative management protocols for children with moyamoya disease.

Protocols for prevention of cerebral ischemic attacks caused by hyperventilation resulting from crying, as observed in perioperative pediatric moyamoya patients, were evaluated. The first protocol involved the use of sedation when staff were setting up the intravenous lines, performing neuroimaging studies, and controlling postoperative pain. The second involved the use of wound-handling techniques designed to ease postoperative wound care; these included steristrip closure, use of paraffin gauze and not using adhesive tapes. We compared 14 and 11 surgical cases handled before and after the protocols were introduced, respectively. The number of patients with perioperative cerebral infarction decreased from 2 to 0. Appropriate sedation reduced the incidence of transient ischemic attacks from 28.6% to 3.7%. The average postoperative hospital stay was similarly reduced, from 21.3 days to 16.1 days, as a consequence of the reduced incidence of complications. It is concluded that the perioperative risks can be minimized when invasive procedures are managed according to our protocols.

Administration, Oral↗

PubMed2003

The effects of protocol and test situation on maximal vs. submaximal cervical motion: medicolegal implications.

The objectives of this study were to examine the influence of the measurement protocol on the range and consistency of cervical motion (CM) in maximal vs. feigned limitation of CM, to explore some cognitive aspects of the feigning performance and to assess the effect of imagined pain vs. financial gain as a stimulus for the submaximal performance. The directions of flexion, extension, right and left rotation and right and left lateral flexion were measured in 26 normal subjects. Four protocols were compared: performance of CM with eyes open vs. eyes closed and testing at either a repetitive (within direction) or random (among directions) order. In each direction three measurements were recorded. Subjects were initially asked to move the head maximally, they were then presented with a vignette describing a fictitious accident involving the neck and were told to feign CM limitation in order to achieve unlawful compensation. In the third part, subjects were instructed to limit CM due to an imagined severe pain applying the repetitive order-eyes open paradigm only. Maximal CM paradigms were associated with significantly larger range ( p=0.0000) and higher consistency ( p=0.0000) compared the feigning paradigms. The eyes open-repetitive order protocol best separated between maximal and feigned performance. It was also indicated that the majority of subjects used the sensation of tension in the neck region as a cue for feigning while attempting to be as consistent as possible. Compared to feigning motivated by financial gain, limitation due to imagined pain resulted in significantly greater CM reductions and lesser consistency. The findings suggest that while feigning of motion impairment is probably based on somato-sensory input, the preferred CM testing protocol should consist of within direction repetitive movements with eyes open.

Adult↗

PubMed2001

A simple, reliable, and fast protocol for thraustochytrid DNA extraction.

DNA extraction of thraustochytrids, common marine unicellular organisms, is usually accomplished by either the cetyltrimethylammonium bromide (CTAB) or proteinase K protocols. A novel lysis buffer protocol for thraustochytrid total DNA extraction is described. The average isolated total DNA is 20 to 40 kb, and DNA samples are suitable for a variety of uses including 18S-ribosomal DNA polymerase chain reaction, restriction enzyme digestions, and amplified fragment length polymorphism analyses. The new protocol is also faster than the other protocols.

Journal Article↗

PubMed2005

Spondylodiscitis after lumbar microdiscectomy: effectiveness of two protocols of intraoperative antibiotic prophylaxis in 1167 cases.

The role of antibiotic prophylaxis in preventing postoperative lumbar spondylodiscitis is still controversial in medical, ethical, economic, and legal terms. The aim of this retrospective study was to evaluate the efficacy of two intraoperative antibiotic prophylaxis protocols in a large series of lumbar microdiscectomies performed in two different neurosurgical centres. We reviewed the outcome of 1167 patients operated on for a lumbar disc herniation with microsurgical technique, in order to detect the incidence of postoperative spondylodiscitis. Group A included 450 patients operated on in a 3-year period in the Neurosurgical Division of the University Hospital of Ancona; group P consisted of 717 patients operated on in a 4-year period in the Neurosurgical Division of the Sandro Pertini Hospital of Rome. In both groups intraoperative antibiotics for prophylaxis were administered, whereas postoperative prophylaxis was not performed. Protocol of group A: single intravenous dose of cefazoline 1 g at induction of general anesthesia and generous washing with saline solution and irrigation with a solution containing rifamicin at the end of microsurgical procedure. Protocol of group P: single-dose of intravenous ampicillin 1000 mg and sulbactam 500 mg at induction of anesthesia and generous irrigation with saline solution at the end of microsurgical procedure. A diagnosis of postoperative spondylodiscitis was made in three out of 450 patients in group A (0.67%) and in 5 out of 717 patients in group P (0.69%). In all cases, treatment consisted of rigid thoraco-lumbar orthesis and 4- to 6-week administration of amoxicillin/clavulanate compound (500/125 mg). The low incidence of postoperative spondylodiscitis obtained with both our protocols seems to confirm that intraoperative antibiotic prophylaxis is associated with the same rate of discitis of prolonged prophylaxis usually still adopted in many centres, but is more advantageous both in terms of welfare and comfort for patients and in economic terms. However, at the moment it is not possible identify the ideal antibiotic for this purpose. It seems to be reasonable to search for the solution through large multicenter prospective studies.

Anti-Bacterial Agents↗

PubMed2003

Protocol for laboratory testing of crude-oil bioremediation products in freshwater conditions.

In 1993, the Environmental Protection Agency, National Risk Management Research Laboratory (EPA, NRMRL), with the National Environmental Technology Application Center (NETAC), developed a protocol for evaluation of bioremediation products in marine environments [18]. The marine protocol was adapted for application in freshwater environments by using a chemically defined medium and an oil-degrading consortium as a positive control. Four products were tested using the modified protocol: two with nutrients and an oleophilic component; one with nutrients, sorbent, and organisms; and one microbial stimulant. A separate experiment evaluated the use of HEPES and MOPSO buffers as replacements for phosphate buffer. The oleophilic nutrient products yielded oil degradation similar to the positive control, with an average alkane removal of 97.1+/-2.3% and an aromatic hydrocarbon removal of 64.8+/-1.2%. The positive control, which received inoculum plus nutrients, demonstrated alkane degradation of 98.9+/-0.1% and aromatic degradation of 52.9+/-0.1%. The sorbent-based product with inoculum failed to demonstrate oil degradation, while the microbial stimulant showed less oil degradation than the positive control. Replacement of phosphate buffer with other buffers had no significant effect on one product's performance. Differences in product performance were easily distinguishable using the protocol, and performance targets for alkane and aromatic hydrocarbon degradation are suggested.

Alkanes↗

PubMed2006

A more cost-effective EMAP benthic macrofaunal sampling protocol.

Benthic macrofaunal sampling protocols in the U.S. Environmental Protection Agency's Environmental Monitoring and Assessment Program (EMAP) are to collect 30 to 50 random benthic macrofauna [defined as animals retained on a 0.5 mm (East and Gulf Coasts, USA) or a 1.0 mm mesh sieve (West Coast, USA)] samples per reporting unit using a 0.044 m(2) (East and Gulf Coasts) or 0.1 m(2) (West Coast) grab. Benthic macrofaunal community conditions in the reporting unit are characterized by cumulative distribution functions (CDFs) on end points of interest, such as number of species (S), abundance (A), and Shannon-Wiener diversity (H'). An EMAP and a companion field study were conducted concurrently in Tillamook Bay (Oregon, USA) to compare the cost effectiveness of benthic macrofauna samples collected using the EMAP West Coast (0.1 m(2) x >or=7 cm deep, 1.0 mm mesh), a 0.01 m(2) x 5 cm deep, 1.0 mm mesh, and a 0.01 m(2) x 5 cm deep, 0.5 mm mesh sampling protocol. Cost was estimated in relative laboratory sample-processing time. Sampling protocols were judged equally effective for EMAP purposes if, after linear transformation to adjust for scale changes in end point distributions, their S, A, and H' CDFs were not significantly different. The 0.01 m(2) x 5 cm deep, 1.0 mm mesh sampling protocol was the most cost effective.

Animals↗

PubMed2006

Issues in conducting cross-cultural research: implementation of an agreed international protocol [corrected] designed by the WHOQOL Group for the conduct of focus groups eliciting the quality of life of older adults.

Multi-centre and cross-cultural research require the use of common protocols if the results are to be either pooled or compared. All too often adherence to protocols is not discussed in reports and where it is reported poor adherence is frequently noted. This paper discusses the use of international guidelines developed by WHOQOL Field Centres to conduct and report focus groups aimed at eliciting key concepts of quality of life among older adults. This was the first step in the development of the WHOQOL-OLD instrument. Although there was overall adherence to the agreed guidelines, there were some differences in the level of reporting, even after participating Field Centres had the opportunity to explain their reports. The reasons for these discrepancies are reported. It is concluded that because of local situations, it is difficult to achieve identical implementation of multi-centre cross-cultural protocols and that the highest standards of auditing are required if findings are to be compared. Suggestions for how such protocols can be improved are given.

Aged↗

PubMed2005

Demonstrating morphometric protocols using polystome marginal hooklet measurements.

The flexible body structure of polystomes (Monogenea: Polystomatidae) encumbers taxonomic classification and species identification. Large intraspecific and limited interspecific variation in the morphology of polystomes further complicates the identification of species. Apart from employing the host-specific nature of the polystomes, taxonomic characterisation relies heavily on the sclerotised skeletal structures, such as the hamuli and the marginal hooklets. The currently accepted measurement system for marginal hooklets appears to be non-optimal and an improved protocol is needed. This paper describes in detail how such a protocol is found by evaluating various sets of measurements statistically in order to identify the most informative combination of parameters. Thirteen measurements of marginal hooklets from 11 different Southern African Polystoma species were taken and evaluated. A new protocol for discriminating between species of Polystoma Zeder, 1800, that employs only three measurements, is proposed. The value of the processes to derive morphometric protocols, as described, is that it is not restricted to a specific taxon, but that it can be amended and applied to any taxonomic grouping.

Animals↗

PubMed2005

Low-molecular-weight-heparins as periprocedural anticoagulation for patients on long-term warfarin therapy: a standardized bridging therapy protocol.

BACKGROUND: Over 2 million patients in North America are on warfarin anticoagulation therapy for prevention of thromboembolism. Suspension of warfarin therapy is often required to prepare patients for invasive procedures or surgeries. To protect these patients against thromboembolism while they are off warfarin, shorter-acting parenteral agents such as low-molecular-weight heparins (LMWHs) are often used. We conducted a retrospective observational study of our anticoagulation clinic patients to assess the safety and efficacy of LMWHs using a standardized protocol for periprocedural anticoagulation therapy. METHODS: We included 69 consecutive patients who required interruption of their long-term warfarin therapy between August 2001 and August 2002, and were deemed by the treating physician to be at high enough risk for perioperative thromboembolism to justify bridging anticoagulation. We used a standard bridging therapy protocol in our anticoagulation clinic. Sixty-six patients received enoxaparin and three patients received tinzaparin for a mean duration of 7.7 days postoperatively. Outcomes were assessed for 30 days post-procedure. Safety outcomes included major bleeding and minor bleeding. Efficacy outcomes included thromboembolic event or death. RESULTS: There were two major bleeding events, one minor bleeding event, and no cases of thromboembolism. Twelve patients experienced some bruising around the injection site. CONCLUSIONS: LMWH administration using our standard outpatient bridging protocol for perioperative anticoagulation appears to be relatively safe and efficacious, offering an alternative to inpatient administration of intravenous unfractionated heparin (UFH). Our study provides additional evidence to the limited published observational data regarding the safety and efficacy of LMWH as bridging therapy in the perioperative and periprocedural setting. Large, multicenter, randomized controlled trials are necessary to fully assess the safety and efficacy of LMWH for perioperative anticoagulation.We conducted a retrospective observational study of 69 consecutive anticoagulation clinic patients on warfarin between August 2001 and August 2002, who were undergoing a procedure or surgery. The study was done to assess the safety and efficacy of an outpatient LMWH bridging protocol. Sixty-six patients received enoxaparin and three patients received tinzaparin for a mean duration of 3 days preoperatively and 7.7 days postoperatively. Outcomes were assessed for 30 days post-procedure. Safety outcomes included major bleeding and minor bleeding. Efficacy outcomes included thromboembolic event or death. There were two major bleeding events, one minor bleeding event, and no cases of thromboembolism. Twelve patients experienced some bruising around the injection site.

Adult↗