Automation of the anthrone assay for carbohydrate concentration determinations.

Reported is the adaptation of a manual polysaccharide assay applicable for glycoconjugate vaccines such as Prevenar to an automated liquid handling system (LHS) for improved performance. The anthrone assay is used for carbohydrate concentration determinations and was scaled to the microtiter plate format with appropriate mixing, dispensing, and measuring operations. Adaptation and development of the LHS platform was performed with both dextran polysaccharides of various sizes and pneumococcal serotype 6A polysaccharide (PnPs 6A). A standard plate configuration was programmed such that the LHS diluted both calibration standards and a test sample multiple times with six replicate preparations per dilution. This extent of replication minimized the effect of any single deviation or delivery error that might have occurred. Analysis of the dextran polymers ranging in size from 214 kDa to 3.755 MDa showed that regardless of polymer chain length the hydrolysis was complete, as evident by uniform concentration measurements. No plate positional absorbance bias was observed; of 12 plates analyzed to examine positional bias the largest deviation observed was 0.02% percent relative standard deviation (%RSD). The high purity dextran also afforded the opportunity to assess LHS accuracy; nine replicate analyses of dextran yielded a mean accuracy of 101% recovery. As for precision, a total of 22 unique analyses were performed on a single lot of PnPs 6A, and the resulting variability was 2.5% RSD. This work demonstrated the capability of a LHS to perform the anthrone assay consistently and a reduced assay cycle time for greater laboratory capacity.

Corpus Cavernosum Abscess Secondary to Traumatic Perforation of Urethral Diverticulum.

Abscess of the corpus cavernosum is a condition that occurs most commonly as a result of penile injection, priapism, sexually transmitted infections, and trauma. The diagnosis of corpus cavernosum abscess is made through imaging, typically computed tomography (CT) or ultrasound. The preferred method of treatment for corpus cavernosum abscess is incision, drainage, and antibiotic therapy. Urethral diverticulum (UD) is defined as a saccular outpouching of the urethral lumen. We present a unique case of corpus cavernosum abscess secondary to perforation of a UD requiring extensive surgical intervention and resulting in long-term complications.

The many facets of cardiovascular magnetic resonance imaging: review of background, clinical utility, and increasing use in the community hospital.

Cardiovascular magnetic resonance imaging (CMR or cardiac MRI) has advanced dramatically in the last ten years and has been proven a reliable and flexible method for cardiac diagnosis. Stress perfusion MRI is a new technique that accurately detects coronary heart disease with high sensitivity and specificity. The role of CMR is also increasing for the evaluation of congestive heart failure and cardiomyopathy. Late gadolinium enhancement highlights areas of myocardial scar and helps predict prognosis. CMR usually complements, but does not fully replace, other diagnostic modalities. Careful patient selection is required to ensure safety in the presence of this powerful magnetic field. MRI gives unique tissue characterization without ionizing radiation. No longer used just for esoteric conditions, CMR is becoming common in the community hospital.

Three-year rabies duration of immunity in dogs following vaccination with a core combination vaccine against canine distemper virus, canine adenovirus type-1, canine parvovirus, and rabies virus.

Thirty-two seronegative pups were vaccinated at 8 weeks of age with modified-live canine distemper virus (CDV), canine adenovirus type-2 (CAV-2), and canine parvovirus (CPV) vaccine and at 12 weeks with a modified-live CDV, CAV-2, CPV, and killed rabies virus vaccine. An additional 31 seronegative pups served as age-matched, nonvaccinated controls. All test dogs were strictly isolated for 3 years after receiving the second vaccination and then were challenged with virulent rabies virus. Clinical signs of rabies were prevented in 28 (88%) of the 32 vaccinated dogs. In contrast, 97% (30 of 31) of the control dogs died of rabies infection. These study results indicated that no immunogenic interference occurred between the modified-live vaccine components and the killed rabies virus component. Furthermore, these results indicated that the rabies component in the test vaccine provided protection against virulent rabies challenge in dogs 12 weeks of age or older for a minimum of 3 years following vaccination.

Three-year duration of immunity in cats following vaccination against feline rhinotracheitis virus, feline calicivirus, and feline panleukopenia virus.

Forty-two seronegative cats received an initial vaccination at 8 weeks of age and a booster vaccination at 12 weeks. All cats were kept in strict isolation for 3 years after the second vaccination and then were challenged with feline calicivirus (FCV) or sequentially challenged with feline rhinotracheitis virus (FRV) followed by feline panleukopenia virus (FPV). For each viral challenge, a separate group of 10 age-matched, nonvaccinated control cats was also challenged. Vaccinated cats showed a statistically significant reduction in virulent FRV-associated clinical signs (P = .015), 100% protection against oral ulcerations associated with FCV infection (P < .001), and 100% protection against disease associated with virulent FPV challenge (P < .005). These results demonstrated that the vaccine provided protection against virulent FRV, FCV, and FPV challenge in cats 8 weeks of age or older for a minimum of 3 years following second vaccination.

The Dual Role of Lipids of the Lipoproteins in Trumenba, a Self-Adjuvanting Vaccine Against Meningococcal Meningitis B Disease.

Trumenba (bivalent rLP2086) is a vaccine licensed for the prevention of meningococcal meningitis disease caused by Neisseria meningitidis serogroup B (NmB) in individuals 10-25 years of age in the USA. The vaccine is composed of two factor H binding protein (fHbp) variants that were recombinantly expressed in Escherichia coli as native lipoproteins: rLP2086-A05 and rLP2086-B01. The vaccine was shown to induce potent bactericidal antibodies against a broad range of NmB isolates expressing fHbp that were different in sequence from the fHbp vaccine antigens. Here, we describe the characterization of the vaccine antigens including the elucidation of their structure which is characterized by two distinct motifs, the polypeptide domain and the N-terminal lipid moiety. In the vaccine formulation, the lipoproteins self-associate to form micelles driven by the hydrophobicity of the lipids and limited by the size of the folded polypeptides. The micelles help to increase the structural stability of the lipoproteins in the absence of bacterial cell walls. Analysis of the lipoproteins in Toll-like receptor (TLR) activation assays revealed their TLR2 agonist activity. This activity was lost with removal of the O-linked fatty acids, similar to removal of all lipids, demonstrating that this moiety plays an adjuvant role in immune activation. The thorough understanding of the structure and function of each moiety of the lipoproteins, as well as their relationship, lays the foundation for identifying critical parameters to guide vaccine development and manufacture.

Testing the theoretical design of a health risk message: reexamining the major tenets of the extended parallel process model.

This study examined the fear control/danger control responses that are predicted by the Extended Parallel Process Model (EPPM). In a campaign designed to inform college students about the symptoms and dangers of meningitis, participants were given either a high-threat/no-efficacy or high-efficacy/no-threat health risk message, thus testing the extreme assumptions of the EPPM. Although the study supports the main predictions of the EPPM in the context of meningitis, the results provide new evidence that only a marginal amount of threat is necessary in a health risk message to move the target audience toward the desired protective measures. In addition, the results also suggest that the messages containing only threat may only scare the target audience further into fear control. Implications and future research are discussed.

Three-year duration of immunity in dogs following vaccination against canine adenovirus type-1, canine parvovirus, and canine distemper virus.

A challenge-of-immunity study was conducted to demonstrate immunity in dogs 3 years after their second vaccination with a new multivalent, modified-live vaccine containing canine adenovirus type 2 (CAV-2), canine parvovirus (CPV), and canine distemper virus (CDV). Twenty-three seronegative pups were vaccinated at 7 and 11 weeks of age. Eighteen seronegative pups, randomized into groups of six dogs, served as challenge controls. Dogs were kept in strict isolation for 3 years following the vaccination and then challenged sequentially with virulent canine adenovirus type 1 (CAV-1), CPV, and CDV. For each viral challenge, a separate group of six control dogs was also challenged. Clinical signs of CAV-1, CPV, and CDV infections were prevented in 100% of vaccinated dogs, demonstrating that the multivalent, modified-live test vaccine provided protection against virulent CAV-1, CPV, and CDV challenge in dogs 7 weeks of age or older for a minimum of 3 years following second vaccination.

Implementation and evaluation of a cognitive-behavioral intervention to prevent problem behavior in a disorganized school.

We assessed the effectiveness of two realizations of cognitive-behavioral instruction in an inner-city middle school with high rates of absenteeism, low staff morale, and chronic low academic achievement. Implementation measures showed that 68% of intended instruction was delivered in the first realization. Self-report measures showed improved school conduct, less victimization in school, and more positive peer associations for the treatment group than for the comparison group at the end of the school year. Treatment students were less likely to leave the school than were comparison students, but were more often absent and tardy. Implementation was poorer in the second realization, and there were no treatment-comparison differences on self-reports or teacher ratings, but treatment students less often left the school. Difficulties in conducting instruction in difficult settings may limit the effectiveness of otherwise efficacious interventions. Specific intervention programs may offer minimal benefits if more basic school improvements are not achieved.