Implementing a patient decision aid, a process evaluation of a large-scale pre- and post-implementation trial.

PURPOSE: Patient decision aids (PtDAs) have been reported to have a positive influence on patients making a health care decision in trials. Nevertheless, post-trial implementation is poor. The aim of this study is to explore patient, clinician, and organizational success factors for implementing a PtDA designed for breast cancer patients, facing a decision on their radiation treatment. METHODS: We performed a process evaluation within a multi-center pre- and post-implementation trial. The PtDA was incorporated as much as possible in the logistics of 13 participating centers. Tracking data were collected on PtDA use. Process characteristics were recorded by both clinicians and patients. A logistic regression method was applied to investigate which process characteristics were significantly related to the probability that patients logged in to the PtDA. RESULTS: 189 patients received the PtDA of whom140 (77%) used the PtDA. If patients received the link via the surgery department they were more likely to use the PtDA (OR 9.77 (1.28-74.51)), compared to patients that received the link via the radiation oncology department. If the report of the multidisciplinary team stated that radiation treatment "had to be discussed with the patient", patients were more likely to use the PtDA (OR 2.29 (1.12-4.71)). Educational level was not related to the probability of PtDA use. CONCLUSIONS: We accomplished a high level of PtDA use. Patients were more likely to use the PtDA if they received the link via the surgery department and if "to be discussed with the patient" was written in the multidisciplinary team report.

Systemic infection and microglia activation: a prospective postmortem study in sepsis patients.

BACKGROUND: Systemic infection is associated with long-term cognitive deficits and functional decline. In this study we hypothesized that severe systemic inflammation leads to a neuroinflammatory response that is characterized by microglial activation, and that these effects might be more pronounced in patients using medication with anticholinergic side-effects. METHODS: Based on the results of a pilot study in 8 patients, we assessed the number of MHC-II and CD-68 positive cells by immunohistochemistry and compared the number of microglia in specific brain regions of 16 well-characterized patients with septic shock and 15 controls. RESULTS: In the pilot study, patients with sepsis tended to have higher density of MHC-II and CD-68 positive microglia in the basal ganglia (putamen, caudate nucleus and globus pallidus) and of MHC-II positive microglia in the hippocampus. In the validation study, patients with sepsis had a significantly higher number of CD-68 positive cells in hippocampus (1.5 fold; p = 0.012), putamen (2.2 fold; p = 0.008) and cerebellum (2.5 fold; p = 0.011) than control patients. The density of MHC-II positive microglia was similar between sepsis and control groups. There was no consistent correlation between microglia counts and anti-cholinergic activity drugs score. CONCLUSION: In patients who die during septic shock, severe systemic inflammation is accompanied by localized and strong upregulation of CD-68 positive microglia, but not of MHC-II positive microglia. We identified regional differences in the brain with increased microglial activation in putamen, hippocampus and cerebellum.

Algorithmic structural segmentation of defective particle systems: a lithium-ion battery study.

We describe a segmentation algorithm that is able to identify defects (cracks, holes and breakages) in particle systems. This information is used to segment image data into individual particles, where each particle and its defects are identified accordingly. We apply the method to particle systems that appear in Li-ion battery electrodes. First, the algorithm is validated using simulated data from a stochastic 3D microstructure model, where we have full information about defects. This allows us to quantify the accuracy of the segmentation result. Then we show that the algorithm can successfully be applied to tomographic image data from real battery anodes and cathodes, which are composed of particle systems with very different morpohological properties. Finally, we show how the results of the segmentation algorithm can be used for structural analysis.

Safety of a non-adjuvanted therapeutic vaccine for the treatment of feline dermatophytosis.

The safety of a non-adjuvanted inactivated fungal vaccine for the treatment of dermatophytosis in cats was investigated in two studies: a controlled laboratory study, and a placebo-controlled double-blind field study with a cross-over design in Europe. In the laboratory study, two groups of 10 cats each were administered an intramuscular twofold overdose, followed by five single 1 ml doses, of either vaccine or control product at 14-day intervals. In the field study, cats were treated with three intramuscular injections of 1 ml vaccine administered at 14-day intervals, as recommended by the manufacturer. A total of 89 cats were enrolled in the field study and divided into two groups to receive either vaccine or placebo for the first three treatments, followed by the opposite product for the final three treatments. The cats enrolled in the two studies were 12 weeks of age or older, as recommended by the manufacturer. All the cats were monitored closely for possible injection site reactions, systemic reactions (including changes in rectal body temperature) and adverse events. The results from both studies showed no significant differences between the vaccinated cats and the control or placebo-treated cats with regard to local or systemic reactions. A few mild to moderate local reactions were noted, but these were evenly distributed between the vaccinated and placebo-treated cats and resolved within a few days. No severe or serious adverse events related to the vaccinations were observed.

Effect of sampling method on the representative recovery of microorganisms from the surfaces of aquacultured finfish.

The objective of this study was to determine if a gentle rinse procedure was equivalent to the combination of excision and homogenization with a stomacher for the relative removal of various microorganisms from finfish fillets. Fillets of hybrid striped bass and rainbow trout were obtained from local markets and sampled using three methods: rinse (R), excision followed by homogenization in a stomacher (S), and homogenization of fillets following a rinse (RS). Microorganisms were enumerated on selective and nonselective media, and randomly selected colonies from aerobic plate counts were identified using MIDI Sherlock and BIOLOG microbial identification systems. Enrichments and selective media were used for the isolation of Listeria monocytogenes, Salmonella spp., and Yersinia enterocolitica. This study confirms previous reports that stomaching is superior to rinsing for enumerating total microbial populations from fish fillets. Rinsing was more effective for rainbow trout than for striped bass. Sampling method did not affect the relative magnitude of plate counts on media selective for aeromonads, pseudomonads, Shewanella, lactic acid bacteria, enterics, and gram-positive cocci. In the compositional analysis of random isolates, R recovered significantly lower fractions of aeromonads than did S or RS, but sampling method did not affect the percent recovery of lactic acid bacteria, pseudomonads, Shewanella, Moraxellaceae, or Cytophaga/Flavobacterium. However, observations suggest that with increased replication, differences among Moraxellaceae, Pseudomonas, and gram positives might be significant. Only one L. monocytogenes colony was isolated, and no Salmonella or Y. enterocolitica, so the effect of sampling method could not be determined for these organisms. Differences in predominant bacterial populations were seen between fish species.

Immunization with Staphylococcus aureus lysate incorporated into microspheres.

Antibiotics are of limited value against Staphylococcus aureus due to development of resistant strains, scar tissue formation, and blockage of ducts due to inflammation. Though macrophages are the predominant cell type in the mammary gland, they are primarily scavenger cells and are not effective against bacteria entering the gland. Neutrophil phagocytosis is the bovine's primary defense against S. aureus mastitis. Attempts to develop vaccines that enhance neutrophil phagocytosis by stimulating production of opsonizing antibodies to S. aureus have met with limited success because of the low immunogenicity of the exopolysaccharide capsule surrounding S. aureus. Staphylococcus aureus can also adhere to and penetrate epithelial tissue. This study was conducted to determine whether lysates of S. aureus encapsulated in biodegradable microspheres would increase the production of opsonizing antibodies to capsule and block adherence. Four groups of four cows each were injected with 1 ml of the respective treatment in the area of the supramammary lymph node and 1 ml in the hip muscle. The treatments were: lysate in NaCl, lysate in Freund's incomplete adjuvant (FICA), lysate in microspheres in NaCl, and lysate in microspheres in FICA. Antigen in microspheres produced a similar antibody response to antigen emulsified in FICA, but to a lesser magnitude. Antigen in microspheres produced antibodies that were more opsonic for neutrophils at 20 and 52 wk postimmunization and inhibited S. aureus adherence to mammary epithelium. Ability to control antigen release and presentation, and the benefit of a single injection for long-term immunity using microspheres warrants additional studies.

Production of antibodies to Staphylococcus aureus serotypes 5, 8, and 336 using poly(DL-lactide-co-glycolide) microspheres.

Staphylococcus aureus is responsible for a major portion of the economic losses due to mastitis. Attempts to produce a vaccine to prevent S. aureus mastitis have been hampered by the low immunogenicity of the polysaccharide, which forms on the surface of the organism when it enters the mammary gland. The polysaccharide inhibits phagocytosis and destruction of the organism by neutrophils. This study was conducted to determine if S. aureus polysaccharide serotypes 5, 8, and 336 conjugated to a protein and incorporated in poly(DL-lactide-co-glycolide) microspheres would enhance the production of opsonizing antibodies to the polysaccharide. Cows were immunized with either polysaccharide conjugates emulsified in Freund's incomplete adjuvant or polysaccharide conjugates encapsulated in poly (DL-lactide-co-glycolide) microspheres emulsified in Freund's incomplete adjuvant. All cows produced sustained antibody titers to the three polysaccharide serotypes. Cows immunized with microspheres had higher antibody titers. Cows in both groups produced increased concentrations of IgG1 and IgG2 antibodies; neither group produced an increase in IgM. Immune sera from cows immunized with conjugates alone increased phagocytosis, which decreased at the end of the study. Sera from cows immunized with conjugates in microspheres increased phagocytosis, which was sustained at the end of the study. Immune sera from both groups decreased bacterial adherence to bovine mammary epithelial cells. These data showed that a single injection of antigen in microspheres produced higher titers and more sustained enhancement of phagocytosis, which could aid in the defense of the cow against S. aureus infections.

The LIM-only protein DRAL/FHL2 binds to the cytoplasmic domain of several alpha and beta integrin chains and is recruited to adhesion complexes.

LIM proteins contain one or more double zinc finger structures (LIM domains) mediating specific contacts between proteins that participate in the formation of multiprotein complexes. We report that the LIM-only protein DRAL/FHL2, with four and a half LIM domains, can associate with alpha(3A), alpha(3B), alpha(7A), and several beta integrin subunits as shown in yeast two-hybrid assays as well as after overexpression in human cells. The amino acid sequence immediately following the conserved membrane-proximal region in the integrin alpha subunits or the C-terminal region with the conserved NXXY motif of the integrin beta subunits are critical for binding DRAL/FHL2. Furthermore, the DRAL/FHL2 associates with itself and with other molecules that bind to the cytoplasmic domain of integrin alpha subunits. Deletion analysis of DRAL/FHL2 revealed that particular LIM domains or LIM domain combinations bind the different proteins. These results, together with the fact that full-length DRAL/FHL2 is found in cell adhesion complexes, suggest that it is an adaptor/docking protein involved in integrin signaling pathways.

Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa.

Evidence is provided that the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase is covalently linked to the fibrous sheath. The fibrous sheath is a typical structure of mammalian spermatozoa surrounding the axoneme in the principal piece of the flagellum. More than 90% of boar sperm glyceraldehyde 3-phosphate dehydrogenase activity is sedimented after cell disintegration by centrifugation. Detergents, different salt concentrations or short term incubation with chymotrypsin do not solubilize the enzyme, whereas digestion with trypsin or elastase does. Short term incubation with trypsin (15 minutes) even resulted in an activation of glyceraldehyde 3-phosphate dehydrogenase. Purification on phenyl-Sepharose yielded a homogeneous glyceraldehyde 3-phosphate dehydrogenase as judged from gel electrophoresis SDS-PAGE and native gradient PAGE. The molecular masses are 41.5 and 238 kDa, respectively, suggesting native glyceraldehyde 3-phosphate dehydrogenase to be a hexamer. Rabbit polyclonal antibodies raised to purified glyceraldehyde 3-phosphate dehydrogenase show a high specificity for mammalian spermatozoal glyceraldehyde 3-phosphate dehydrogenase, while other proteins of boar spermatozoa or the muscle glyceraldehyde 3-phosphate dehydrogenase are not labelled. Immunogold staining performed in a post-embedding procedure reveals the localization of glyceraldehyde 3-phosphate dehydrogenase along the fibrous sheath in spermatozoa of boar, bull, rat, stallion and man. Other structures such as the cell membrane, dense fibres, the axoneme or the mitochondria are free of label. During the process of sperm maturation, most of the cytoplasm of the sperm midpiece is removed as droplets during the passage through the epididymis. The labelling of this cytoplasm, in immature boar spermatozoa and in the droplets, indicates that glyceraldehyde 3-phosphate dehydrogenase is completely removed from the midpiece during sperm maturation in the epididymis. The inverse compartmentation of the glycolytic enzyme and mitochondria in the mammalian sperm flagella suggests that ATP-production in the principal piece mainly occurs by glycolysis and in the midpiece by respiration.

Prevention of nonspecific binding of immunoglobulin to Staphylococcus aureus protein A in ELISA assays.

The Fc region of IgG of most mammals binds protein A on S. aureus resulting in high backgrounds when measuring specific antibodies to S. aureus in the ELISA. Removal of protein A from S. aureus or modification of the Ig Fc to prevent binding to protein A could affect specific antibody binding. We compared effects of blockage of Fc binding to protein A with purified protein A to trypsin removal of protein A from S. aureus, on specific antibody binding. When NMS was incubated without and with protein A (0 microgram, 50 micrograms, 200 micrograms and 400 micrograms) and high protein A Cowan I was the bound S. aureus antigen in the ELISA, absorbance OD405 was 0.769, 0.240, 0.224 and 0.210 +/- SE 0.026. When mouse Mab (IgG1, kappa) to bovine IgA was incubated without and with protein A (400 micrograms) prior to reaction with bovine IgA in the ELISA, absorbance was 0.645 and 0.639, indicating protein A had no effect on specific antibody binding. To determine the effect of trypsin on specific binding, Becker S. aureus was trypsin treated before linking it to microtiter wells. When Mab (IgM) to Becker (Nelles et al., Infect. Immun. (1985) 49, 14) was incubated with protein A (400 micrograms) before use in the ELISA, trypsin treatment of Becker resulted in reduced specific antibody activity (untreated Becker = 1.306, trypsin treated Becker = 0.331). These results suggest that purified protein A can be used to block nonspecific binding via Fc of Ig to S. aureus, thus avoiding trypsin denaturation of surface antigens.

Prevalence and Distribution of Campylobacter jejuni and Campylobacter coli in Retail Meats.

Nine cooperating laboratories, distributed throughout the United States, determined the interlaboratory reproducibility of a sensitive, selective method for isolation of Campylobacter jejuni and Campylobacter coli from foods, and determined the prevalence and distribution of the organism in retail meats. A double-blind inoculated/recovery experiment demonstrated the ability to detect two cells of C. jejuni and C. coli per g of meat at a rate of 96% among the cooperating laboratories. However, a 7.5% false-positive rate for the presumptive detection of the organism was also reported. Samples of ground beef, beef flank steak, lamb stew meat, broiler chicken, pork sausage (without antimicrobials), and pork chops were selected to assess the presence of campylobacters. Each cooperator purchased five of each of the above samples from the refrigerated case of two retail outlets at quarterly intervals throughout the year. A total of 2,160 retail samples were analyzed for the presence of C. jejuni and C. coli . Results indicated that about 30% of the 360 chickens sampled yielded the organism. Analysis of 1,800 red meat products yielded campylobacters at a rate of about 5.1%. Pork samples yielded C. coli and other meats yielded C. jejuni . Higher numbers of isolations from the red meats were made during June and September (8.6%) as compared with December and March (4.2%). These results provide a baseline, for the prevalence of campylobacters in these selected foods, and also support epidemiologic data associating mishandled foods of animal origin as a potential vehicle in human gastroenteritis.

Selected enrichment broths for recovery of Campylobacter jejuni from foods.

We attempted to shorten the required time for enrichment broth culture for the isolation of Campylobacter jejuni. Enrichment broths described by Doyle and Roman and Park and Stankiewicz and one developed during this study were compared for ability to isolate C. jejuni from raw chicken carcasses. Our medium was a modification of that of Doyle and Roman with the addition of filter-sterilized FBP (0.2% ferrous sulfate, 0.025% sodium metabisulfite, 0.05% sodium pyruvate), 0.1% sodium lauryl sulfate, and 0.075% agar. Initially, laboratory strains were employed in the development of this medium. Subsequently, an indigenous load of C. jejuni obtained from chickens was used to compare media. Isolation rate comparisons were as follows: direct plating, 40%; Doyle and Roman broth, 45% at 7 h and 61% at 16 h; Park and Stankiewicz broth, 53% at 7 h and 60% at 16 h; our broth, 48% at 7 h and 50% at 16 h. In addition to having the highest isolation rate, the enrichment broth of Doyle and Roman showed greatest selectivity. Our inoculation method of indigenous bacteria provided a controlled means for comparison of isolation procedures.

Suitability of the ASM-2 standard test of the National Committee for Clinical Laboratory Standards for evaluation of antimicrobial disk potency.

Standard disks of 25 antimicrobial agents were prepared and tested at three levels of potency (67, 100, and 150% of labeled quantity). The method used was a modification of the approved standard M2-A2 of the National Committee for Clinical Laboratory Standards. Forty-seven susceptibility tests were performed at each potency level by using one to three test organisms. Labeled-potency (100%) disks were within accuracy limits for 85% of the tests and were within daily control limits for 94% of the tests. All susceptibility test data for labeled disks, however, were considered acceptable. The majority (63%) of mean zone diameter data for labeled-content disks were in the upper-one-third percentile of accuracy control limits. A significant proportion (91%) of low-potency disks and considerably fewer (34%) of the high-potency disks were found acceptable when daily control limits were applied. Of most concern are those antimicrobial agents whose low-potency disks approach the lower region of control limits. Zone diameter data from standard disks in the range of potency levels tested suggest that control ranges are excessive for some antimicrobial agent-test organism combinations.

Isolation and identification of ropy bacteria in raw milk.

Approximately 4.2% of 4,000 Maryland-Virginia raw milk tanker samples developed ropiness when incubated at 10 degrees C. Of the 56 bacterial isolates 30 were identified by species. Klebsiella oxytoca and Pseudomonas aeruginosa were isolated most frequently. Other ropy isolates were identified as Pseudomonas spp., Chromobacterium, Flavobacterium multivorum, presumptive Yersinia pestis, Enterobacter agglomerans, Klebsiella pneumoniae, and Pasteurella-Actinobacter spp. Six of the Klebsiella oxytoca isolates were mesophilic (optimum temperatures of 32.0 to 37.8 degrees C) with two isolates having psychrotrophic tendencies (optimum temperature of 26.8 degrees C). All Pseudomonas aeruginosa isolates appeared to be psychrotropic in their temperature requirements (optimum temperature of 23.0 to 31.0 degrees C). Klebsiella oxytoca was significant in preliminary development of the ropy condition. All Klebsiella oxytoca isolates developed ropiness within 24 h. The Pseudomonas spp. and Klebsiella pneumoniae isolates required at long as 7 days to develop detectable ropiness at 10 degrees C. A recommended Klebsiella oxytoca differentiation agar is presented as a rapid screening method during outbreaks where Klebsiella oxytoca is the organism of significance.

Selective method for the isolation of Sporolactobacillus from food and environment sources.

A detection procedure was developed in which a modified Lactobacilli MRS medium was used to enrich for Sporolactobacillus from a variety of foods, feed soil and environmental samples. Each sample was rinsed with 50 ml of a modified Lactobacilli MRS broth containing 1.0% (w/v) alpha-methyl glucoside 0.1% (w/v) potassium sorbate, 0.00224% (w/v) bromocresol green indicator, adjusted to pH 5.5 with acetic acid and incubated at 37 degrees C for 7 d under 5% CO2. Volumes of 2 ml from each sample were heat shocked at 80 degrees C for 5 min and 0.1 ml spread onto plates of Lactobacilli MRS agar (Difco), pH 5.5 and APT agar (BBL), pH 5.5. Plates were incubated for 5 d and suspect colonies were tested for catalase production, benzidine reaction, nitrate reduction, motility and Gram stain reaction. This method was demonstrated to be selective for Sporolactobacillus.

Influence of lactose concentration of milk and yogurt on growth rate of rats.

Growth rates of rats fed lyophilized diets of yogurt, milk, 100% lactose-hydrolyzed milk, and 100% lactose-hydrolyzed yogurt were compared. No differences were significant in weight gains between the animals fed lactose-hydrolyzed diets over 3 wk. Further more, rats fed the lactose-hydrolyzed diets had significantly larger weight gains during the 1st wk than those fed unhydrolyzed diets. This may have been from the deleterious effect of lactose, which was in concentrations of 43% in milk and 36% in yogurt. However, by the 2nd wk, rats fed yogurt had growth rates similar to those fed lactose-hydrolyzed diets, and by the 3rd wk, no differences of growth rates were significant. Continued consumption of high concentrations of lactose improved digestion of lactose. Yogurt fermentation produced growth benefit as compared to the milk from which it was made by reducing the lactose content and by supplying microbial lactase activity.

Microbiology of ultrahigh temperature milk.

Ultrahigh temperature thermal processing can sterilize milk. Potential energy savings of a commercially sterile, aseptically packaged, nonrefrigerated milk provide the incentive for eventual introduction of the product in the United States. Attention should be directed to raw milk quality, processing parameters, quality control tests, and thermal inactivation data for spores in the ultrahigh temperature range.

Effect of 13-Nle-motilin on small intestinal transit time in healthy subjects.

In 6 female subjects without gastrointestinal diseases, 13-norleucine motilin (13-Nle-M) synthetic and biological analogue of the duodenal polypeptide, motilin, caused acceleration of intestinal transit time. Intravenous infusion of 0.4 mug/kg-h of 13-Nle-M reduced mean transit time by 50 percent. No side effects occurred during infusion of the polypeptide. Radiographic appearance of small intestinal peristalsis and mucosal relief was not influenced by 13-Nle-M.

[Multilocular tumors of small intestine (author's transl)]. / Multilokulärer Dünndarmtumor

Tumors of small intestine can be recognized in time and evaluated histologically by means of roentgenographic and endoscopical-bioptic methods, respectively. Among radiologic techniques, the double contrast examination of the small intestine has especially high diagnostic value. On the other hand, selective angiography reveals bleeding of accompanying small intestinal tumors. Out of 23 patients with these tumors -- diagnosed in the Medical Department of the Erlangen University from 1968 to July 1974 -- there was one case with multilocular occurrence; the diagnostic efficiency of double contrast technique proved to be particularly suitable in this patient.