Inter-rater agreement and reliability of thoracic ultrasonographic findings in feedlot calves, with or without naturally occurring bronchopneumonia.

BACKGROUND: Thoracic ultrasonography (TUS) can be used to assess the extent and severity of lung lesions associated with bronchopneumonia (BP) in feedlot cattle. HYPOTHESIS/OBJECTIVES: To assess inter-rater agreement and reliability of TUS findings in feedlot cattle, with or without naturally occurring BP. ANIMALS: Feedlot steers with (n = 210) or without (n = 107) clinical signs of BP that were assessed by TUS in a previous case-control study. METHODS: A random sample of 50 TUS videos (16-s duration) were scored by 6 raters with various levels of TUS expertise. Lung consolidation, comet tail artifacts, pleural irregularity and effusion were scored. Inter-rater agreement was assessed using raw percentage of agreement (Pa), Cohen's and Fleiss' Kappa (κ), and Gwet agreement coefficient (AC1). Intra-class correlation (ICC) was determined for variables with continuous measurements (mixed factorial design). RESULTS: Median (interquartile range [IQR]) Pa were 0.84 (0.80-0.89), 0.82 (0.80-0.87), 0.62 (0.53-0.67), and 0.82 (0.75-0.86) for presence of lung consolidation, comet tails, pleural irregularity, and pleural effusion, respectively. For the same lesions, Fleiss κ (95% confidence intervals [CI]) were 0.67 (0.49-0.86), 0.56 (0.33-0.80), 0.20 (-0.05 to 0.44), and 0.36 (0.10-0.61), respectively. AC1 were 0.68 (0.51-0.86), 0.73 (0.58-0.89), 0.21 (-0.01 to 0.44), and 0.71 (0.51-0.92), respectively. Moderate reliability was found among raters for all quantitative variables (ICC ranged from 0.52 to 0.70). CONCLUSIONS AND CLINICAL IMPORTANCE: Inter-rater agreement was good for presence of lung consolidation, comet tails and pleural effusion (based on Pa and AC1) but was slight to poor for pleural irregularity.

New chemical and radiochemical routes to [18F]Rho6G-DEG-F, a delocalized lipophilic cation for myocardial perfusion imaging with PET.

New chemical and radiochemical syntheses are described for the preparation of [18F]Rho6G-DEG-F, an 18F-labeled analogue of the fluoresecent dye rhodamine 6G, which has shown promise as myocardidal perfusion imaging agent. Tosylated precursors of [18F]Rho6G-DEG-F amenable to 18F-labeling were obtained either through a two-step synthesis from rhodamine 6G lactone (33% yield), or in one step from rhodamine 575 (64% yield), then purified by preparative C18 chromatography. Manual synthesis of [18F]Rho6G-DEG-F was achieved in a single radiochemical step from either the tosylate salt or the tosylate/formate double salt in DMSO under standard nucleophillic aliphatic 18F-fluorination conditions (K[18F]F/K2CO3/Kryptofix 2.2.2.). Incorporation of the [18F]F- was found to be satisfactory (≥34% by TLC), despite the protic character of the precursor molecules. [18F]Rho6G-DEG-F was manually synthesized in final decay-corrected radiochemical yields of 11-26% (tosylate salt) and 9-21% (tosylate/formate double salt). The protocol was transferred to an automated synthesis unit, where the product was obtained in 3-9% radiochemical yield (n=3) decay corrected to start-of-synthesis, >99% radiochemical purity, and a molar activity of 122-267 GBq/µmol (3.3-7.2 Ci/µmol).

Data in support for the measurement of serum 25-hydroxyvitamin D (25OHD) by tandem mass spectrometry.

This article provides data and a method related to a research paper entitled "Assessing vitamin D nutritional status: is capillary blood adequate?" (Jensen et al., 2016) [1]. Circulating 25OHD, the accepted biomarker of the vitamin D nutritional status, is routinely measured by automated immunoassays, that although may be performed in hospital central laboratories, often suffer from a lack of specificity with regards to the different vitamin D metabolites, "Measurement of circulating 25-hydroxyvitamin D: a historical review" (Le Goff et al., 2015) [2]. Mass spectrometry offers this specificity. This article describes the performance of an in-house tandem mass spectrometry method for the individual measurement of 25OHD3, 25OHD2 and 3-épi-25OHD3.

Assessing vitamin D nutritional status: Is capillary blood adequate?

BACKGROUND: Venous blood is the usual sample for measuring various biomarkers, including 25-hydroxyvitamin D (25OHD). However, it can prove challenging in infants and young children. Hence the finger-prick capillary collection is an alternative, being a relatively simple procedure perceived to be less invasive. We elected to validate the use of capillary blood sampling for 25OHD quantification by liquid chromatography tandem-mass spectrometry (LC/MS-MS). METHODS: Venous and capillary blood samples were simultaneously collected from 15 preschool-aged children with asthma 10days after receiving 100,000IU of vitamin-D3 or placebo and 20 apparently healthy adult volunteers. 25OHD was measured by an in-house LC/MS-MS method. RESULTS: The venous 25OHD values varied between 23 and 255nmol/l. The venous and capillary blood total 25OHD concentrations highly correlated (r(2)=0.9963). The mean difference (bias) of capillary blood 25OHD compared to venous blood was 2.0 (95% CI: -7.5, 11.5) nmol/l. CONCLUSION: Our study demonstrates excellent agreement with no evidence of a clinically important bias between venous and capillary serum 25OHD concentrations measured by LC/MS-MS over a wide range of values. Under those conditions, capillary blood is therefore adequate for the measurement of 25OHD.

Definition of prepartum hyperketonemia in dairy goats.

A prospective cohort study was conducted on 1,081 dairy goats from 10 commercial herds in Québec (Canada) to define prepartum hyperketonemia based on optimal blood ß-hydroxybutyrate acid threshold values for the early prediction of pregnancy toxemia (PT) and mortality in late-gestation dairy goats. All pregnant goats had blood sampled weekly during the last 5wk of pregnancy. The blood was analyzed directly on the farm for ß-hydroxybutyrate acid quantification using a Precision Xtra meter (Abbott Diabetes Care, Saint-Laurent, QC, Canada). Body condition scores on the lumbar region and sternum were noted. Each goat was classified as being at low (n=973) or high risk (n=108) of having PT by producers based on a standardized definition. The optimal threshold for predicting a PT diagnosis or mortality for each week before kidding was determined based on the highest sum of sensitivity and specificity. The association between hyperketonemia and subsequent PT was tested using a multivariable logistic regression model considering hyperketonemia at wk 4 prepartum, litter size, and body condition score at wk 4 prepartum as covariates, and herd and parturition cohort as random effects. The association between mortality and hyperketonemia was also tested using a logistic regression model accounting for the presence or absence of treatment during the last month of pregnancy. The hyperketonemia definition based on PT varied between ≥0.4 and ≥0.9mmol/L during the last 5wk prepartum. Goats affected by hyperketonemia at wk 4 prepartum and with a large litter size (≥3 fetuses) had 2.1 and 40.5 times the odds, respectively, of subsequent PT than other goats. Hyperketonemia definitions based on mortality varied between ≥0.6 and ≥1.4mmol/L during the last 4wk prepartum, and was ≥1.7mmol/L during the first week postpartum. Goats affected by hyperketonemia and treated by producers had 3.4 and 11.8 times the odds, respectively, of subsequent mortality than did other goats. These results showed that prepartum hyperketonemia could be defined in dairy goats using subsequent risks of PT or mortality during the last month of pregnancy.

Respiratory pathogens in Québec dairy calves and their relationship with clinical status, lung consolidation, and average daily gain.

BACKGROUND: Bovine respiratory disease (BRD) is 1 of the 2 most important causes of morbidity and mortality in dairy calves. Surprisingly, field data are scant concerning the prevalence of respiratory pathogens involved in BRD in preweaned dairy calves, especially in small herds. OBJECTIVES: To identify the main respiratory pathogens isolated from calves in Québec dairy herds with a high incidence of BRD, and to determine if there is an association between the presence of these pathogens and clinical signs of pneumonia, lung consolidation, or average daily gain. ANIMALS: Cross-sectional study using a convenience sample of 95 preweaned dairy calves from 11 dairy herds. METHODS: At enrollment, calves were weighed, clinically examined, swabbed (nasal and nasopharyngeal), and lung ultrasonography was performed. One month later, all calves were reweighed. RESULTS: Twenty-two calves had clinical BRD and 49 had ultrasonographic evidence of lung consolidation. Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni were isolated in 54, 17, and 12 calves, respectively. Mycoplasma bovis was identified by PCR testing or culture in 19 calves, and 78 calves were found to be positive for Mycoplasma spp. Bovine coronavirus was detected in 38 calves and bovine respiratory syncytial virus in 1. Only the presence of M. bovis was associated with higher odds of clinical signs, lung consolidation, and lower average daily gain. CONCLUSIONS AND CLINICAL IMPORTANCE: Results suggested that nasopharyngeal carriage of M. bovis was detrimental to health and growth of dairy calves in small herds with a high incidence of BRD.

Preoperative cow-side lactatemia measurement predicts negative outcome in Holstein dairy cattle with right abomasal disorders.

The objectives of the current study were (1) to determine the gain in prognostic accuracy of preoperative l-lactate concentration (LAC) measured on farm on cows with right displaced abomasum (RDA) or abomasal volvulus (AV) for predicting negative outcome; and (2) to suggest clinically relevant thresholds for such use. A cohort of 102 cows with on-farm surgical diagnostic of RDA or AV was obtained from June 2009 through December 2011. Blood was drawn from coccygeal vessels before surgery and plasma LAC was immediately measured by using a portable clinical analyzer. Dairy producers were interviewed by phone 30 d following surgery and the outcome was determined: a positive outcome if the owner was satisfied of the overall evolution 30 d postoperatively, and a negative outcome if the cow was culled, died, or if the owner reported being unsatisfied 30 d postoperatively. The area under the curve of the receiver operating characteristic curve for LAC was 0.92 and was significantly greater than the area under the curve of the receiver operating characteristic curve of heart rate (HR; 0.77), indicating that LAC, in general, performed better than HR to predict a negative outcome. Furthermore, the ability to predict a negative outcome was significantly improved when LAC measurement was considered in addition to the already available HR data (area under the curve: 0.93 and 95% confidence interval: 0.87, 0.99). Important inflection points of the misclassification cost term function were noted at thresholds of 2 and 6 mmol/L, suggesting the potential utility of these cut-points. The 2 and 6 mmol/L thresholds had a sensitivity, specificity, positive predictive value, and negative predictive value for predicting a negative outcome of 76.2, 82.7, 53.3, and 93.1%, and of 28.6, 97.5, 75, and 84%, respectively. In terms of clinical interpretation, LAC ≤2 mmol/L appeared to be a good indicator of positive outcome and could be used to support a surgical treatment decision. The treatment decision for cows with LAC between 2 and 6 mmol/L, however, would depend on the economic context and the owner's attitude to risk in regard to potential return on its investment. Finally, performing a surgical correction on commercial cows with RDA or AV and a LAC ≥6 mmol/L appeared to be unjustified and these animals should be culled based on their high probability of negative outcome.

Comparison of thoracic auscultation, clinical score, and ultrasonography as indicators of bovine respiratory disease in preweaned dairy calves.

BACKGROUND: The diagnostic tools for bovine respiratory disease diagnosis include clinical inspection, thoracic auscultation, and ultrasonography. HYPOTHESIS: Thoracic auscultation and clinical examination have limitations in the detection of lung consolidation in dairy calves. ANIMALS: Prospective cohort of 106 preweaned calves from 13 different dairy herds (10 with a history of active bovine respiratory disease (BRD) in calves and 3 without suspected BRD problems). METHODS: Each preweaned calf was clinically inspected using the Wisconsin calf respiratory scoring chart (CRSC) and treatment history was noted. Systematic thoracic auscultation and ultrasonography then were performed, the latter focusing on lung consolidation. Mortality was recorded over a 30-day period. RESULTS: A total of 56 of 106 calves had ultrasonographic evidence of lung consolidation. The sensitivity of thoracic auscultation to detect consolidation was 5.9% (range, 0-16.7%). Only 41.1% (23/33) of calves with consolidated lungs had been treated previously by the producers. When adding CRSC and previous BRD treatment by the producer, sensitivity of detection increased to 71.4% (40/56). The area under the receiver operating characteristics curve was 0.809 (95% CI, 0.721-0.879) for the number of areas within the lungs with consolidation and 0.743 (95% CI, 0.648-0.823) for the maximal depth of consolidation as predictors of death within 1 month after examination. These were not significantly different (P = .06). CONCLUSIONS AND CLINICAL IMPORTANCE: This study shows that thoracic auscultation is of limited value in diagnosing lung consolidation in calves. Ultrasonographic assessment of the thorax could be a useful tool to assess BRD detection efficiency on dairy farms.

Short communication: ultrasonographic assessment of the thorax as a fast technique to assess pulmonary lesions in dairy calves with bovine respiratory disease.

The aim of this study was to assess inter- and intraoperator agreement when assessing lung consolidation secondary to bovine respiratory disease (BRD) by thoracic ultrasonography. Ten calves were blindly assessed by 3 operators with varying expertise in thoracic ultrasound to look for lung consolidation and the presence of comet-tail artifacts (COMT). Systematic ultrasonography of the thorax was performed using an 18-site per side assessment with a linear 8.5-MHz probe. The status of the calves [healthy (n=4) vs. treated for BRD (n=6)] was not known by the operators. The interoperator kappa agreement for detecting consolidation was moderate to almost perfect (from 0.6 to 1.0) depending on the operator's experience (diagnosis of consolidation if depth ≥1cm). The intraclass correlation coefficient for consistency was 0.71 for a single measurement and 0.88 for average measurement. The intraclass correlation coefficient for agreement was 0.73 for single measurements and 0.89 for average measurements. These values were considered good for single measurements and excellent for average measurements. Systematic ultrasonography of the thorax can be used routinely to assess lung consolidation in dairy calves and can therefore be of importance, especially for assessment of subclinical BRD.

Short communication: evaluation of the accuracy of an electronic on-farm test to quantify blood ß-hydroxybutyrate concentration in dairy goats.

The objective of this cross-sectional study was to validate the accuracy of a hand-held electronic on-farm test (Precision Xtra) for quantifying the blood ß-hydroxybutyrate (BHBA) concentration in dairy goats. A total of 114 dairy goats from 3 commercial herds were sampled once for blood in the jugular vein between 1mo before and 2mo after parturition. Blood samples were centrifuged to harvest serum and sera were sent to the Animal Health Laboratory of the Université de Montréal for quantification of BHBA concentration (gold standard). Laboratory BHBA values were between 0.1 and 3.7mmol/L. Precision Xtra values were compared with gold standard values; Pearson correlation coefficient was 0.98 and coefficient of determination was 0.95. Overall, these results suggested that Precision Xtra provides excellent accuracy for measuring blood BHBA concentration in dairy goats compared with the gold standard test.

Ultrasonographic assessment of standing laparotomy wound healing in dairy cows.

The surgical wounds of 40 cows with a standing clean laparotomy were assessed 10-15 days after surgery. The abdominal wall thickness on the upper (w1), the middle (w2) and the lower (w3) third of the wound (and the contralateral flank n1, n2, and n3), the presence of edema, cavitary lesion, capsulated lesion, or visualization of shadow artifact in the wound were assessed ultrasonographically. On day 30, wound healing status (normal or surgical site infection (SSI)) was determined. Five out of 39 cows (12.8%) had SSI. A significant positive correlation was found between w3 and edema (p<0.01; r=0.44), w3/n3 and edema (p<0.05; r=0.32), w3 and pain on wound palpation (p<0.01; r=0.41); w3/n3 and the presence of pain (p<0.002; r=0.49) and w2/n2 and pain when palpating the wound (p<0.05; r=0.33). No association was found between the ultrasonographic evidence of edema and SSI (p=0.10), cavitary lesion and SSI (p=0.65), encapsulated lesion and SSI (p=1), and shadow artefact and SSI (p=1). The wound ultrasonography 10-15 days after surgery was not predictive of abnormal wound healing.

The effect of stimulus type and background noise on hearing abilities of the round goby Neogobius melanostomus.

The auditory abilities of the round goby Neogobius melanostomus were quantified using auditory evoked potential recordings, using tone bursts and conspecific call stimuli. Fish were tested over a range of sizes to assess effects of growth on hearing ability. Tests were also run with and without background noise to assess the potential effects of masking in a natural setting. Neogobius melanostomus detected tone bursts from 100 to 600 Hz with no clear best frequency in the pressure domain but were most sensitive to 100 Hz tone stimuli when examined in terms of particle acceleration. Responses to a portion of the N. melanostomus call occurred at a significantly lower threshold than responses to pure tone stimulation. There was no effect of size on N. melanostomus hearing ability, perhaps due to growth of the otolith keeping pace with growth of the auditory epithelium. Neogobius melanostomus were masked by both ambient noise and white noise, but not until sound pressure levels were relatively high, having a 5-10 dB threshold shift at noise levels of 150 dB re 1 µPa and higher but not at lower noise levels.

Use of ichip for high-throughput in situ cultivation of "uncultivable" microbial species.

One of the oldest unresolved microbiological phenomena is why only a small fraction of the diverse microbiological population grows on artificial media. The "uncultivable" microbial majority arguably represents our planet's largest unexplored pool of biological and chemical novelty. Previously we showed that species from this pool could be grown inside diffusion chambers incubated in situ, likely because diffusion provides microorganisms with their naturally occurring growth factors. Here we utilize this approach and develop a novel high-throughput platform for parallel cultivation and isolation of previously uncultivated microbial species from a variety of environments. We have designed and tested an isolation chip (ichip) composed of several hundred miniature diffusion chambers, each inoculated with a single environmental cell. We show that microbial recovery in the ichip exceeds manyfold that afforded by standard cultivation, and the grown species are of significant phylogenetic novelty. The new method allows access to a large and diverse array of previously inaccessible microorganisms and is well suited for both fundamental and applied research.

Neuromuscular stimulation therapy after incomplete spinal cord injury promotes recovery of interlimb coordination during locomotion.

The mechanisms underlying the effects of neuromuscular electrical stimulation (NMES) induced repetitive limb movement therapy after incomplete spinal cord injury (iSCI) are unknown. This study establishes the capability of using therapeutic NMES in rodents with iSCI and evaluates its ability to promote recovery of interlimb control during locomotion. Ten adult female Long Evans rats received thoracic spinal contusion injuries (T9; 156 +/- 9.52 Kdyne). 7 days post-recovery, 6/10 animals received NMES therapy for 15 min/day for 5 days, via electrodes implanted bilaterally into hip flexors and extensors. Six intact animals served as controls. Motor function was evaluated using the BBB locomotor scale for the first 6 days and on 14th day post-injury. 3D kinematic analysis of treadmill walking was performed on day 14 post-injury. Rodents receiving NMES therapy exhibited improved interlimb coordination in control of the hip joint, which was the specific NMES target. Symmetry indices improved significantly in the therapy group. Additionally, injured rodents receiving therapy more consistently displayed a high percentage of 1:1 coordinated steps, and more consistently achieved proper hindlimb touchdown timing. These results suggest that NMES techniques could provide an effective therapeutic tool for neuromotor treatment following iSCI.

Partial androgen suppression consequent to increased secretion of adrenal androgens in a patient with prostate cancer treated with long-acting GnRH agonists.

We present a case report of a patient with prostate cancer who failed to demonstrate consistent testosterone suppression to castration levels and incomplete suppression of serum prostate-specific antigen, although treated with gonadotropin releasing hormone agonists for 48 months. Serum dehydroepiandrosterone, dehydroepiandrosterone sulphate, as well as the androgen metabolite, androsterone glucuronide, were elevated compared to the other patients. The present data suggest that those prostate cancer patients who have even marginally elevated adrenal androgens may especially benefit from combined androgen blockade.

Prolonged gestation in two Holstein cows: transabdominal ultrasonographic findings in late pregnancy and pathologic findings in the fetuses.

Prolonged pregnancy in cattle is a rare condition that is commonly accompanied with fetal adenohypophyseal hypoplasia. The final diagnosis is often challenging as breeding date error remains possible in virtually all situations. In this report, fetal wellbeing assessment is depicted in two Holstein cows suffered from prolonged pregnancy. Transabdominal ultrasonographic findings were compatible with healthy non-stressed fetuses. The clinical and pathologic findings encountered in both fetuses expelled after corticosteroid induction of parturition consisted of adenohypophyseal aplasia and hydronephrosis. Other nervous system anomalies (hydrocephaly or holocephaly) were observed. Tetralogy of Fallot was also found in one calf.

Bioavailability and metabolism of oral and percutaneous dehydroepiandrosterone in postmenopausal women.

To study the bioavailability of dehydroepiandrosterone (DHEA) administered by the oral and percutaneous routes, three groups of 12 postmenopausal women aged 60-70 years received two capsules of 50mg of DHEA orally before breakfast daily for 14 days or applied 4 g of a 10% DHEA cream or gel at the same time of the day on a 30 cm x 30 cm surface area on the thighs. Detailed serial blood sampling over 24h was performed following 1st and 14th DHEA administration for measurement of DHEA and nine of its metabolites by liquid chromatography tandem mass spectrometry (LC-MS/MS) or gas chromatography mass spectrometry (GC-MS). Serum levels of estrone (E1) and estradiol (E2) did not change following DHEA administration by any of the three formulations, while serum androstenedione (4-dione), testosterone, DHEA sulfate (DHEA-S), E(1)-S, androsterone glucuronide (ADT-G) and 3alpha-androstanediol-G (3alpha-diol-G), increased in all cases, the effect on these parameters being more important after oral than percutaneous administration due to the metabolism of DHEA into these metabolites in the gastrointestinal tract and liver. No qualitative differences in DHEA metabolism are observed between the oral and percutaneous routes of DHEA administration while the levels of all steroids remain on a plateau during the 24h period during chronic percutaneous DHEA administration. The present data show that DHEA is transformed into active androgens and estrogens in peripheral intracrine tissues with no or minimal release of the active steroids E(1), E(2) or testosterone in the circulation. Moreover, DHEA is preferentially transformed into androgens rather than into estrogens. Most importantly, the present data show that changes in serum DHEA following oral or percutaneous DHEA administration are not a valid parameter of DHEA action since the increase in serum DHEA is at least 100% greater than the increase in the formation of active androgens and estrogens and thus much higher than the potential physiological effects.

Is dehydroepiandrosterone a hormone?

Dehydroepiandrosterone (DHEA) is not a hormone but it is a very important prohormone secreted in large amounts by the adrenals in humans and other primates, but not in lower species. It is secreted in larger quantities than cortisol and is present in the blood at concentrations only second to cholesterol. All the enzymes required to transform DHEA into androgens and/or estrogens are expressed in a cell-specific manner in a large series of peripheral target tissues, thus permitting all androgen-sensitive and estrogen-sensitive tissues to make locally and control the intracellular levels of sex steroids according to local needs. This new field of endocrinology has been called intracrinology. In women, after menopause, all estrogens and almost all androgens are made locally in peripheral tissues from DHEA which indirectly exerts effects, among others, on bone formation, adiposity, muscle, insulin and glucose metabolism, skin, libido and well-being. In men, where the secretion of androgens by the testicles continues for life, the contribution of DHEA to androgens has been best evaluated in the prostate where about 50% of androgens are made locally from DHEA. Such knowledge has led to the development of combined androgen blockade (CAB), a treatment which adds a pure anti-androgen to medical (GnRH agonist) or surgical castration in order to block the access of the androgens made locally to the androgen receptor. In fact, CAB has been the first treatment demonstrated to prolong life in advanced prostate cancer while recent data indicate that it can permit long-term control and probably cure in at least 90% of cases of localized prostate cancer. The new field of intracrinology or local formation of sex steroids from DHEA in target tissues has permitted major advances in the treatment of the two most frequent cancers, namely breast and prostate cancer, while its potential use as a physiological HRT could well provide a physiological balance of androgens and estrogens, thus offering exciting possibilities for women's health at menopause.