Influence of a HTR2B Stop Codon on Glucagon Homeostasis and Glucose Excursion in Non-Diabetic Men.

Limited data are available about the role of the serotonin 2B (5-HT2B) receptor in the function of human islets. This study aimed to test whether the 5-HT2B receptor contributes to glucose, insulin, and glucagon homeostasis in humans, utilizing a hereditary loss-of-function gene mutation in the receptor, which causes a 50% reduction in the production of the receptor protein in heterozygotes. This clinical study enrolled participants recruited by newspaper advertisements and from mental status examinations. A cohort of participants from a young Finnish founder population composed of 68 non-diabetic males with a mean age of 30 was divided into groups for comparison based on being a 5-HT2B receptor loss-of-function gene mutation (HTR2B Q20*) heterozygote carrier (n=11) or not (n=57). Serum levels of glucose, insulin, and glucagon were measured in a 5 h oral glucose tolerance test using a 75 g glucose challenge. Insulin resistance, insulin sensitivity, and beta cell activity were calculated using the homeostasis model assessment (HOMA2) and whole body insulin sensitivity index (WBISI), as well as the ratio of glucagon to insulin was noted. The areas under the curves (AUCs) were also determined. Concentrations of the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) were measured in cerebrospinal fluid (CSF). Covariate adjusted mean score comparisons were applied. Lower glucagon secretion and decreased glucose excursion were observed among HTR2B Q20* carriers as compared with individuals who were homozygotes for the wild-type Q20 allele (controls). No differences in insulin secretion, beta cell activity, insulin resistance, or insulin sensitivity were observed. The glucagon to insulin ratio differed between the HTR2B Q20* carriers and controls. CSF levels of 5-HIAA were similar between groups. Our findings indicate that the 5-HT2B receptor may contribute to the regulation of human glucagon and glucose homeostasis and the interplay between glucagon and insulin secretion.

Clinical Research Abstracts of the British Equine Veterinary Association Congress 2015.

REASONS FOR PERFORMING STUDY: Infiltrative disease of the intestine is an important cause of weight loss in the horse. Infiltration of inflammatory or neoplastic cells into the intestinal wall and intestinal fibrosis cause changes in the integrity of the intestinal wall. This may lead to altered intestinal permeability which can be measured using the contrast medium iohexol. OBJECTIVES: To determine if iohexol intestinal permeability, as evaluated by serum iohexol concentration, could be used to differentiate between healthy horses and horses with infiltrative disease of the large colon. STUDY DESIGN: Prospective non-randomised controlled clinical trial. METHODS: Six healthy adult horses and 4 horses with chronic infiltrative disease of the large colon were used in the study. Infiltrative disease was confirmed on post mortem in all cases, and included alimentary lymphoma and mycobacterial granulomatous enterocolitis. Following a 16-h fast, each horse was dosed with 1.0 ml/kg bwt of iohexol as a 10% solution via nasogastric intubation. Blood samples were collected at 0, 30, 60, 120, 180, 240, 300, 360, 420 and 480 min after dosing. Iohexol concentration was determined using HPLC-UV and the differences between the groups were analysed with a repeated measures ANOVA. RESULTS: There was a statistically significant difference in iohexol serum concentration between the diseased and nondiseased horses (P = 0.001). The overall difference in the mean iohexol concentration between the 2 groups was 6.07 (95% CI 3.19-8.96) µg/ml, however there appeared to be a trend towards increasing difference at later time points (240, 300, 360 min). CONCLUSIONS: The iohexol permeability test has potential as a diagnostic tool for estimation of intestinal permeability in horses with infiltrative intestinal disease. Further studies are warranted to determine whether the test can be used to determine the site of intestinal pathology, predict the prognosis and potentially evaluate the response to treatment. ACKNOWLEDGEMENTS: The authors thank Professor Riitta-Mari Tulamo and Professor Thomas Spillmann and the staff of Equine College Ypäjä and the University of Helsinki Equine Teaching Hospital. The cooperation of horse owners is gratefully acknowledged. Ethical animal research: The study protocol was approved by the National Animal Experiment Board of Finland (Eläinkoelautakunta ELLA, Request for Animal Experiments, ref. no. ESAVI-2010-06567/Ym-23). For client-owned animals, owner informed consent was obtained. SOURCE OF FUNDING: This study was funded by the Faculty of Veterinary Medicine, University of Helsinki, Finland. Competing interests: None declared.

Uncovering cortical MEG responses to listened audiobook stories.

Naturalistic stimuli, such as normal speech and narratives, are opening up intriguing prospects in neuroscience, especially when merging neuroimaging with machine learning methodology. Here we propose a task-optimized spatial filtering strategy for uncovering individual magnetoencephalographic (MEG) responses to audiobook stories. Ten subjects listened to 1-h-long recording once, as well as to 48 repetitions of a 1-min-long speech passage. Employing response replicability as statistical validity and utilizing unsupervised learning methods, we trained spatial filters that were able to generalize over datasets of an individual. For this blind-signal-separation (BSS) task, we derived a version of multi-set similarity-constrained canonical correlation analysis (SimCCA) that theoretically provides maximal signal-to-noise ratio (SNR) in this setting. Irrespective of significant noise in unaveraged MEG traces, the method successfully uncovered feasible time courses up to ~120 Hz, with the most prominent signals below 20 Hz. Individual trial-to-trial correlations of such time courses reached the level of 0.55 (median 0.33 in the group) at ~0.5 Hz, with considerable variation between subjects. By this filtering, the SNR increased up to 20 times. In comparison, independent component analysis (ICA) or principal component analysis (PCA) did not improve SNR notably. The validity of the extracted brain signals was further assessed by inspecting their associations with the stimulus, as well as by mapping the contributing cortical signal sources. The results indicate that the proposed methodology effectively reduces noise in MEG recordings to that extent that brain responses can be seen to nonrecurring audiobook stories. The study paves the way for applications aiming at accurately modeling the stimulus-response-relationship by tackling the response variability, as well as for real-time monitoring of brain signals of individuals in naturalistic experimental conditions.

Intersubject consistency of cortical MEG signals during movie viewing.

According to recent functional magnetic resonance imaging (fMRI) studies, spectators of a movie may share similar spatiotemporal patterns of brain activity. We aimed to extend these findings of intersubject correlation to temporally accurate single-trial magnetoencephalography (MEG). A silent 15-min black-and-white movie was shown to eight subjects twice. We adopted a spatial filtering model and estimated its parameter values by using multi-set canonical correlation analysis (M-CCA) so that the intersubject correlation was maximized. The procedure resulted in multiple (mutually uncorrelated) time-courses with statistically significant intersubject correlations at frequencies below 10 Hz; the maximum correlation was 0.28 ± 0.075 in the ≤1 Hz band. Moreover, the 24-Hz frame rate elicited steady-state responses with statistically significant intersubject correlations up to 0.29 ± 0.12. To assess the brain origin of the across-subjects correlated signals, the time-courses were correlated with minimum-norm source current estimates (MNEs) projected to the cortex. The time series implied across-subjects synchronous activity in the early visual, posterior and inferior parietal, lateral temporo-occipital, and motor cortices, and in the superior temporal sulcus (STS) bilaterally. These findings demonstrate the capability of the proposed methodology to uncover cortical MEG signatures from single-trial signals that are consistent across spectators of a movie.

Quality of bulk tank milk samples from Danish dairy herds based on real-time polymerase chain reaction identification of mastitis pathogens.

Results of a commercial real-time PCR analysis for 11 mastitis pathogens from bulk tank milk (BTM) samples from all 4,258 Danish dairy herds in November 2009 to January 2010 were compared with somatic cell count (SCC) and total bacteria count (TBC) estimates in BTM. For Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis, a low real-time PCR cycle threshold (Ct) value (corresponding to high bacterial DNA quantity) was correlated with higher SCC and higher TBC. For Staphylococcus aureus, low Ct values were correlated only with higher SCC. For the environmental mastitis pathogens Klebsiella spp., Enterococcus spp., and Escherichia coli, low Ct values had a correlation with higher TBC. Staphylococcus spp. were found in the BTM from all herds, Strep. uberis in 95%, Staph. aureus in 91%, and Strep. dysgalactiae in 86%, whereas E. coli, Klebsiella, and Strep. agalactiae were found in 61, 13, and 7% of the herds. It is concluded that the real-time PCR used provides results that are related to the milk quality in the herds. Real-time PCR can be used in the same way as culture for monitoring BTM samples, and is especially useful for bacteria with low prevalence (e.g., Strep. agalactiae).

Population studies of 16 bovine STR loci for forensic purposes.

As a consequence of the close integration of cattle into the food chain of humans, forensically relevant cases involving cattle (Bos taurus) DNA analysis are common. However, scientific publications reporting the information content of the commonly used bovine short tandem repeat (STR) loci remains scarce. Population studies were performed for 16 polymorphic STR loci (BM1818, BM1824, BM2113, CSRM60, CSSM66, ETH3, ETH10, ETH225, HAUT27, ILSTS006, INRA023, SPS115, TGLA53, TGLA122, TGLA126, and TGLA227) including 4,162 randomly selected cattle representing 20 distinct breeds. The power of parental exclusion, expected and observed heterozygosity, probability of identity, and non-amplifying ("null") allele frequencies were calculated. Major differences existed in the information content between different cattle breeds. The selection of 16 STR loci, partially recommended by International Society for Animal Genetics as the minimum standard needed for bovine STR typing, was sufficient for forensic analysis. Furthermore, the efficacy of the loci was assessed in assigning unknown individuals to the correct breed based on genotype data. The individual assignment tests provided excellent success in several breeds.

Addendum to 'Continuous assessment of nasal airflow resistance by adaptive modeling'.

This addendum adds to the analysis of 'Continuous assessment of nasal airflow resistance by adaptive modeling' (Seppänen et al 2009 Physiol. Meas. 30 1197­209). The technical repeatability tests of the new nasal resistance measurement system presented here show that the resistance values remained very stable during two successive measurements indicating excellent repeatability.

Field comparison of real-time polymerase chain reaction and bacterial culture for identification of bovine mastitis bacteria.

Fast and reliable identification of the microorganisms causing mastitis is important for management of the disease and for targeting antimicrobial treatment. Methods based on PCR are being used increasingly in mastitis diagnostics. Comprehensive field comparisons of PCR and traditional milk bacteriology have not been available. The results of a PCR kit capable of detecting 11 important etiological agents of mastitis directly from milk in 4h were compared with those of conventional bacterial culture (48h). In total, 1,000 quarter milk samples were taken from cows with clinical or subclinical mastitis, or from clinically healthy quarters with low somatic cell count (SCC). Bacterial culture identified udder pathogens in 600/780 (77%) of the clinical samples, whereas PCR identified bacteria in 691/780 (89%) of the clinical samples. The PCR analysis detected major pathogens in a large number of clinical samples that were negative for the species in culture. These included 53 samples positive for Staphylococcus aureus by PCR, but negative by culture. A total of 137 samples from clinical mastitis, 5 samples from subclinical mastitis, and 1 sample from a healthy quarter were positive for 3 or more bacterial species in PCR, whereas culture identified 3 or more species in 60 samples from clinical mastitis. Culture identified a species not targeted by the PCR test in 44 samples from clinical mastitis and in 9 samples from subclinical mastitis. Low SCC samples provided a small number of positive results both in culture (4/93; 4.3%) and by PCR (7/93; 7.5%). In conclusion, the PCR kit provided several benefits over conventional culture, including speed, automated interpretation of results, and increased sensitivity. This kit holds much promise as a tool to complement traditional methods in identification of pathogens. In conventional mastitis bacteriology, a sample with 3 or more species is considered contaminated, and resampling of the cow is recommended. Further study is required to investigate how high sensitivity of PCR and its quantitative features can be applied to improve separation of relevant udder pathogens from likely contaminants in samples where multiple species are detected. Furthermore, increasing the number of species targeted by the PCR test would be advantageous.

Parathyroid scintigraphy protocols in Finland in 2010. Results of the query and current status.

UNLABELLED: The GOAL of this study was to describe current clinical parathyroid scintigraphy (PS) protocols in Finland. METHODS: all departments of nuclear medicine in Finland were contacted, and instructions regarding PS were requested. RESULTS: instructions regarding PS were received from all of the departments that perform PS (n = 19). Seven hundred and sixty-nine PS studies were performed in 2009 (between 7 and 209 per hospital). Three methods of PS were used. The dual-phase method with (99m)Tc-sestamibi is used in seven hospitals, the dual-tracer method with (123)I/(99m)Tc-sestamibi in eleven, and (99m)TcO4/(99m)Tc-sestamibi in one hospital. The activities of 99mTc-sestamibi, 123I and (99m)TcO4 were 150-800 MBq, 10-30 MBq and 50 MBq, respectively. The anterior image with parallel-hole collimators, the anterior image with pinhole collimator, the oblique angles with pinhole collimator, SPECT and hybrid CT with SPECT were acquired in 84%, 26%, 16%, 63%, and 42% of the hospitals, respectively. Because the imaging techniques were combined, one to four acquisitions were performed per patient. Scatter and attenuation correction were used in five protocols. A correction for crosstalk of (123)I and (99m)Tc gamma energies was not used, but the amount of crosstalk was decreased by using narrow or asymmetric energy windows in all dual isotope protocols. CONCLUSION: 19 hospitals used 18 different study protocols. Thus, significant variability exists in the current practice of PS in Finland. The protocols should be tested with known phantoms to determine any differences in sensitivities for detecting small active structures. Further studies with phantoms are needed to determine the optimal imaging techniques. The results of these phantom studies will provide guidelines for proposing national recommendations for PS.

Continuous assessment of nasal airflow resistance by adaptive modeling.

A method to assess nasal airflow resistance is presented that provides a continuous resistance value and applies a novel minimally obtrusive measurement technique. Instead of calculating the resistance once for each breathing cycle conventionally, it is calculated for each signal sample at any sampling frequency. The continuous pressure recording is produced with a nasopharyngeal catheter inserted 8 cm deep along the floor of the other nasal cavity and the flow recording is produced with respiratory effort bands. A least-mean-square (LMS) extension for the resistance model of Broms is developed that dynamically adapts to the time-varying characteristics of the nasal functioning and produces the continuous resistance values. Experimental results are shown that demonstrate the uniqueness and applicability of the new technique in assessing quickly changing resistance in a histamine/xylometatsolin challenges, the differences between normal and symptomatic patients, and the effect of nasal treatment of patients.

Real-time polymerase chain reaction-based identification of bacteria in milk samples from bovine clinical mastitis with no growth in conventional culturing.

In more than 30% of milk samples from clinical and subclinical bovine mastitis, bacteria fail to grow even after 48 h of conventional culture. The "no-growth" samples are problematic for mastitis laboratories, veterinarians, and dairy producers. This study provides the first investigation of the bacteriological etiology of such samples, using a real-time PCR-based commercial reagent kit. The assay targets the DNA of the 11 most common bacterial species or groups in mastitis and the staphylococcal blaZ gene (responsible for penicillin resistance) and can identify and quantify bacterial cells even if dead or growth-inhibited. A study was made of 79 mastitic milk samples with no-growth bacteria in conventional culture, originating from cows with clinical mastitis. Of the 79 samples, 34 (43%) were positive for 1 (32 samples) or 2 (2 samples) of the target bacteria. The positive findings included 11 Staphylococcus spp. (staphylococci other than Staphylococcus aureus), 10 Streptococcus uberis, 2 Streptococcus dysgalactiae, 6 Corynebacterium bovis, 3 Staph. aureus, 1 Escherichia coli, 1 Enterococcus, and 1 Arcanobacterium pyogenes. The positive samples contained as many as 10(3) to 10(7) bacterial genome copies per milliliter of milk. This study demonstrates that in nearly half of the clinical mastitis cases in which conventional culture failed to detect bacteria, mastitis pathogens were still present, often in substantial quantities. The clearly elevated N-acetyl-beta-d-glucosaminidase activity values of the milk samples, together with clinical signs of the infected cows and quarters, confirmed the diagnosis of clinical mastitis and indicated that real-time, PCR-based bacterial findings are able to reveal bacteriological etiology. We conclude that all common mastitis bacteria can occur in large quantities in clinical mastitis samples that exhibit no growth in conventional culture, and that the real-time PCR assay is a useful tool for bacteriological diagnosis of such milk samples. Low bacterial concentration is commonly speculated to explain the no-growth milk samples. This hypothesis is not supported by the results of the current study.

Analytical specificity and sensitivity of a real-time polymerase chain reaction assay for identification of bovine mastitis pathogens.

Intramammary infection (IMI), also known as mastitis, is the most frequently occurring and economically the most important infectious disease in dairy cattle. This study provides a validation of the analytical specificity and sensitivity of a real-time PCR-based assay that identifies 11 major pathogen species or species groups responsible for IMI, and a gene coding for staphylococcal beta-lactamase production (penicillin resistance). Altogether, 643 culture isolates originating from clinical bovine mastitis, human, and companion animal samples were analyzed using the assay. The isolates represented 83 different species, groups, or families, and originated from 6 countries in Europe and North America. The analytical specificity and sensitivity of the assay was 100% in bacterial and beta-lactamase identification across all isolates originating from bovine mastitis (n = 454). When considering the entire culture collection (including also the isolates originating from human and companion animal samples), 4 Streptococcus pyogenes, 1 Streptococcus salivarius, and 1 Streptococcus sanguis strain of human origin were identified as Streptococcus uberis, and 3 Shigella spp. strains were identified as Escherichia coli, decreasing specificity to 99% in Strep. uberis and to 99.5% in E. coli. These false-positive results were confirmed by sequencing of the 16S rRNA gene. Specificity and sensitivity remained at 100% for all other bacterial targets across the entire culture collection. In conclusion, the real-time PCR assay shows excellent analytical accuracy and holds much promise for use in routine bovine IMI testing programs. This study provides the basis for evaluating the assay's diagnostic performance against the conventional bacterial culture method in clinical field trials using mastitis milk samples.

Extended release of adenovirus from silica implants in vitro and in vivo.

Despite promising preclinical results, the clinical benefits of cancer gene therapy have been modest heretofore. The main obstacle continues to be the level and persistence of gene delivery to sufficiently large areas of the tumor. One approach for overcoming this might entail extended local virus release. We studied the utility of silica gel monoliths for delivery of adenovirus to advanced orthotopic gastric and pancreatic cancer tumors. Initially, the biochemical properties of the silica-virus matrix were studied and nearly linear release as a function of time was detected. Virus stayed infective for weeks at +37 degrees C and months at +4 degrees C, which may facilitate storage and distribution. In vivo, extended release of functional replication deficient and also replication-competent, capsid-modified oncolytic viruses was seen. Treatment of mice with pancreatic cancer doubled their survival (P<0.001). Also, silica gel-based delivery slowed the development of antiadenovirus antibodies.

Effects of noise and filtering on SVD-based morphological parameters of the T wave in the ECG.

Singular value decomposition (SVD) based electrocardiogram (ECG) morphology analysis is a novel method in the assessment of subtle abnormalities in the T wave morphology of 12-lead ECG. As various types of noise contaminate the ECG signal and create a bias for the morphological analyses, this study was designed to estimate the effects of noise on the SVD method in an experimental setup. Ideal signals were generated by filtering real ECG signals several times with the Savitzky-Golay filter. Random and real noise samples were superimposed on the ideal signals. The noisy signals were filtered with a power line interference filter combined with the Savitzky-Golay or the wavelet filter. Results show that noise increased both the dipolar and non-dipolar components significantly unless filtering was applied. R-TWR (relative T wave residuum) and A-TWR (absolute T wave residuum) were four to eight times higher in noisy signals. The experiments with patient data demonstrated that certain types of noise may even lead to erroneous classification of patients. Filtering brings the median values closer to the correct ones and decreases significantly the variance of the values of parameters.

Metabolism of OR-1896, a metabolite of levosimendan, in rats and humans.

OR-1896 is a pharmacologically active, long-lived metabolite of levosimendan. In the current study, the metabolism of (14)C-labelled OR-1896 was investigated in six healthy men after intravenous infusion over 10 min and in male rats after an intravenous bolus dose. In human plasma, the only (14)C-compounds detected were (14)C-OR-1896 and its deacetylated form, (14)C-OR-1855, in varying proportions in different subjects. In rat plasma >93% of radioactivity was associated with OR-1896. Radioactivity was mainly excreted to urine in both rats (about 69% of the dose) and humans (about 87% of the dose). OR-1896 was a major urinary compound in both humans and rats. Another major human metabolite was hypothesized as N-conjugated OR-1855. Other human and rat urinary biotransformation products were characterized as N-hydroxylated OR-1896 and N-hydroxylated OR-1855, as well as glucuronide or sulphate conjugates of N-hydroxyl OR-1896. The main difference between rat and human metabolism was a lower amount of OR-1855-related metabolites in the rats. In human faecal homogenates, only OR-1896 and OR-1855 were detected, whereas rat faecal metabolite profile was similar to that in urine.

Comparison of tests for detection of beta-lactamase-producing staphylococci.

Penicillin resistance identification tests are important in veterinary medicine. Six enzyme assays and a PCR test were compared for the detection of beta-lactamase production or the beta-lactamase gene in 175 staphylococcal isolates. We conclude that the PCR test and two nitrocefin-based assays can be recommended for routine clinical use.

Spectral properties of rotating electrons in quantum dots and their relation to quantum Hall liquids.

The exact diagonalization technique is used to study many-particle properties of interacting electrons with spin, confined in a two-dimensional harmonic potential. The single-particle basis is limited to the lowest Landau level. The results are analysed as a function of the total angular momentum of the system. Only at angular momenta corresponding to the filling factors 1, 1/3, 1/5, etc is the system fully polarized. The lowest energy states exhibit spin waves, domains, and localization, depending on the angular momentum. Vortices exist only at excited polarized states. The high angular momentum limit shows localization of electrons and separation of the charge and spin excitations.

Isolation by distance within a river system: genetic population structuring of Atlantic salmon, Salmo salar, in tributaries of the Varzuga River in northwest Russia.

An important issue for designing any conservation programme aimed at preserving genetic diversity is estimation of the scale at which genetic structuring occurs. Additional relevant factors include distinguishing whether or not population structuring is expected to be stable as predicted by the member-vagrant hypothesis, or alternatively, whether populations are more prone to local extinction-recolonization processes, as predicted by the metapopulation evolutionary model. In this study, the population genetic structure of Atlantic salmon from 11 locations within or nearby the Varzuga River tributary system was assessed using 17 microsatellites. Mantel tests and spatial autocorrelation analyses revealed a significant isolation-by-distance signal within the tributary system as well as a negative association between the level of genetic diversity and waterway distance from the river mouth, indicating that dispersal is less likely to occur to populations deep in the tributary system. Individual-level spatial autocorrelation analyses indicated that the majority of migration occurred between populations situated within 20 km of each other. The relatively high level of genetic structuring and significant isolation-by-distance signal observed in the Varzuga tributaries are concordant with the predictions of the member-vagrant evolutionary model. However, one subpopulation in particular revealed signs of instability which may be due to its location in the tidal zone, or due to the fact that it is more affected by human impacts. The results suggest that preservation of a number of spawning sites spaced throughout the tributary system is recommendable for ensuring sustainable fishing tourism in the river.