Pathogens associated with pleuritic pig lungs at an abattoir in Queensland Australia.

OBJECTIVE: Pleurisy in pigs has economic impacts in the production stage and at slaughter. This study sought to establish if some micro-organisms can be found in high numbers in lungs with pleurisy by assessing batches of pigs at an abattoir in Queensland Australia. DESIGN: Samples of lung (including trachea/bronchus and lymph nodes) from a maximum of 5 pleurisy affected pigs were collected from 46 batches of pigs representing 46 Queensland farms. PROCEDURE: Pleurisy-affected lung areas were cultured by traditional bacteriological methods and bacteria quantified by plate scores. Additionally, tracheal or bronchial swabs and apical lobe fluid were tested for Mycoplasma hyopneumoniae DNA and the superior tracheobronchial lymph nodes were tested for porcine circovirus type 2 DNA by polymerase chain reaction (PCR). All apparently significant bacteria were identified via PCR or sequencing. Typing was undertaken on some of the bacterial isolates. RESULTS: The most prevalent pathogens were M. hyopneumoniae, Streptococcus suis and Porcine Circovirus type 2, being found in 34, 38 and 31 batches, respectively. Other bacteria found were Actinobacillus species (29 batches), Pasteurella multocida (24 batches), Mycoplasma flocculare (9 batches), Actinobacillus pleuropneumoniae (7 batches), Mycoplasma hyorhinis (4 batches), Bisgaard Taxon 10 (1 batch), Glaesserella parasuis (1 batch), Streptococcus minor (1 batch) and Streptococcus porcinus (1 batch). Most batches had more than one bacterial species. CONCLUSION: The high percentage of batches infected with S. suis (83%), M. hyopneumoniae (74%) and PCV2 (70%) and clustering by a batch of these pathogens, as well as the presence of many secondary pathogens, suggests synergy between these organisms may have resulted in pleurisy.

A comparison of the reproducibility of two cine-derived strain software programmes in disease states.

BACKGROUND: Systolic strain and peak-early diastolic strain rate (PEDSR) measure subclinical cardiac dysfunction. These parameters can be derived from cardiovascular magnetic resonance (CMR) cine images using new software packages, but the comparative test-retest reproducibility of these software in disease states is unknown. This study compared the test-retest reproducibility of strain measures derived from two software packages (feature-tracking software (FT) and tissue-tracking (TT)) in disease populations with preserved ejection fractions. METHODS: This was a prospective study of 10 patients with aortic stenosis (AS), 10 haemodialysis patients and 10 diabetic patients at 1.5 and 3-Tesla. 30 subjects underwent test-retest reproducibility scans of global circumferential strain (GCS), global longitudinal strain (GLS), circumferential-PEDSR and longitudinal-PEDSR calculated using TT and FT software. RESULTS: Test-retest reproducibility of GCS and GLS were similar for FT and TT across patient groups. Coefficient of variability (CoV) for FT-derived GCS 8.1%, 5% and 7.9% for AS, diabetic and haemodialysis patients, compared to 3.3%, 9.2% and 5.4% for TT-derived GCS, with CoV for FT-derived GLS 8%, 6.4% and 8.2% for AS, diabetic and haemodialysis patients, compared to 5.3%, 4.8% and 7% for TT-derived GLS). Reproducibility of FT-derived circumferential and longitudinal-PEDSR was worse than TT-derived circumferential and longitudinal-PEDSR (CoV for FT-derived circumferential-PEDSR 18.2%, 18% and 17.4% for AS, diabetic and haemodialysis patients, compared to 6.1%, 11.7% and 11% for TT-derived circumferential-PEDSR with CoV for FT-derived longitudinal PEDSR 18.2%, 18.9%, 18.3% for AS, diabetic and haemodialysis patients, compared to 8.9%, 9.1% and 11.4% for TT-derived longitudinal-PEDSR). Bland-Altman analysis revealed no systematic bias with tighter limits of agreement for TT-derived strain measures. CONCLUSIONS: Reproducibility of GCS and GLS are excellent with FT and TT software across diseases. TT had superior test-retest reproducibility for quantification of longitudinal and circumferential-PEDSR than FT-derived PEDSR across diseases.

Higher antineutrophil cytoplasmic antibody (C-ANCA) titers are associated with increased overall healthcare use in patients with sinonasal manifestations of granulomatosis with polyangiitis (GPA).

BACKGROUND: Granulomatosis with polyangiitis (GPA) is an autoimmune disease characterized by necrotizing granulomatous airway inflammation and vasculitis. Sinonasal involvement occurs in more than 80% cases, with antineutrophil cytoplasmic antibody (C-ANCA) titers used as a marker of disease severity. The purpose of this study was to determine whether C-ANCA levels impact radiographic findings and healthcare use in patients with sinonasal GPA. METHODS: A retrospective review was performed on GPA patients evaluated in a multidisciplinary rheumatologic/otolaryngologic clinic from 2008 to 2013. Data were collected with respect to age, gender, clinical presentation, C-ANCA titers, Lund-Mackay (LM) scores, surgical interventions, and healthcare use, the latter of which were determined by assessing the number of rheumatology/otolaryngology clinic visits, computed tomography (CT) scans, and email/telephone encounters. RESULTS: A total of 44 patients were identified, 11 male and 33 female. Sinonasal manifestations were evident in 70.4%, with chronic rhinosinusitis (CRS) (41.9%), septal perforation (38.7%), and crusting (32.2%) the most common findings. No significant differences in number of CT scans (p = 0.10) or mean LM scores (p = 0.47) were found between patients with more than or equal to 1:80 and less than 1:80 C-ANCA titers, respectively. However, overall healthcare use was increased in the more than or equal to 1:80 C-ANCA group (n = 28) compared with less than 1:80 (n = 16), with a significantly greater number of rheumatologic/otolaryngologic encounters (mean 121 versus 69.2, p = 0.03) noted. When otolaryngologic healthcare use was specifically examined, the average number of encounters was also higher in more than or equal to 1:80 C-ANCA patients (31.9 versus 22.9), but this difference was not statistically significant (p = 0.16). CONCLUSION: Sinonasal GPA patients with presenting C-ANCA titers more than or equal to 1:80 demonstrated significantly greater overall healthcare use than their lower C-ANCA level counterparts (less than 1:80). However, no significant differences in otolaryngology resource use or LM scores were evident between the two titer groups.

Negatively charged, dye labeled-dideoxynucleotides for "direct-load" DNA sequencing.

A four-color set of negatively charged, single dye as well as energy transfer dye labeled-ddNTPs were synthesized and evaluated in combination with a novel polymerase in a "direct-load" DNA sequencing, obviating the laborious and time consuming post-reaction work-up.

Synthesis and energy transfer efficiency of FRET terminators derived from different linkers.

A number of different energy transfer dye labeled-cassettes were synthesized using aminoacid based trifunctional linkers and coupled to the propargylamino-substituted dideoxynucleoside-5'-triphosphates (ddNTPs). These terminators were evaluated for their energy transfer efficiency and DNA sequencing potential using thermostable DNA polymerase.

Efficient incorporation of positively charged 2', 3'-dideoxynucleoside-5'-triphosphates by DNA polymerases and their application in 'direct-load' DNA sequencing.

A series of charge-modified, dye-labeled 2', 3'-dideoxynucleoside-5'-triphosphates have been synthesized and evaluated as reagents for dye-terminator DNA sequencing. Unlike the commonly used dye-labeled terminators, these terminators possess a net positive charge and migrate in the opposite direction to dye-labeled Sanger fragments during electrophoresis. Post-sequencing reaction purification is not required to remove unreacted nucleotide or associated breakdown products prior to electrophoresis. Thus, DNA sequencing reaction mixtures can be loaded directly onto a separating medium such as a sequencing gel. The charge-modified nucleotides have also been shown to be more efficiently incorporated by a number of DNA polymerases than regular dye-labeled dideoxynucleotide terminators or indeed normal dideoxynucleoside-5'-triphosphates.

Synthesis and application of charge-modified dye-labeled dideoxynucleoside-5'-triphosphates to 'direct-load' DNA sequencing.

A novel series of charge-modified, dye-labeled 2',3'-dideoxynucleoside-triphosphate terminators were synthesized and evaluated as reagents for DNA sequencing. These terminators possess an advantage over existing reagents in that no purification is required to remove unreacted nucleotide or associated breakdown products prior to electrophoretic separation of the sequencing fragments. This obviates the need for a time consuming post-reaction work up, allowing direct loading of DNA sequencing reaction mixtures onto a slab gel. Thermo Sequenase II DNA polymerase poorly incorporates the charge-modified terminators compared with regular dye-labeled terminators. However, extending the linker arm between dye and nucleotide and using a mutant form of a related DNA polymerase can in part mitigate the decrease in substrate efficiency. We also present evidence that these charge-modified terminators can relieve gel compression artefacts when used with dGTP in sequencing reactions.

Self-reported desire to improve colposcopic impressions.

BACKGROUND: Women who participate in cervical cancer screening programs must have access to high quality colposcopy services when their cytology test are abnormal. The purpose of this project is to evaluate colposcopic services currently available in New Hampshire and whether colposcopy providers are willing to network to maintain and improve their colposcopic pattern recognition skills in order to improve their colposcopic correlations. METHODS: A survey was mailed to 1314 providers throughout New Hampshire. The survey ascertained the extent of current colposcopic services in New Hampshire through practitioner and practice demographics, the cervical procedures performed, the self-reported colposcopy skill level, and the self-reported quality of past colposcopic education. It also measured interest in networking with others to improve their colposcopic pattern recognition skills. RESULTS: The survey response rate was 62%. 145 of the 810 respondents (18%) are currently performing colposcopies, indicating a statewide potential colposcopy accommodation rate of 3.5 to 7 women per month per colposcopist. 57% of the physician assistants, 59% of the family physicians, 75% of the gynecologists and 100% of the nurse practitioners were interested in enhancing their colposcopic pattern recognition skills by networking through quarterly meetings. CONCLUSIONS: The crude accommodation rate for colposcopy appears sufficient for the women of New Hampshire although the geographic distribution of the colposcopists is unknown. A majority of the colposcopists were interested in networking to improve their colposcopic pattern recognition skills, which could improve patient care.