Reliability of environmental sampling to quantify Mycobacterium avium subspecies paratuberculosis on California free-stall dairies.

The reliability of environmental sampling to quantify Mycobacterium avium ssp. paratuberculosis (MAP) based on collector and time was evaluated. Fecal slurry samples were collected using a standardized protocol simultaneously by 2 collectors of different experience levels. Samples were collected from 30 cow pens on 4 dairies every other day on 3 occasions while cow movements between pens were minimal. The 4 study herds had moderate MAP seroprevalence and were housed in free-stall dairies in central California. Results of testing the environmental samples for MAP using PCR and culture were strongly correlated. The reliability of environmental sampling simultaneously by different collectors as estimated by the intraclass correlation coefficient was excellent (81%) for PCR and good (67%) for culture and may justify comparison of quantitative results of samples collected by different investigators. The reliability of environmental sampling over a 5-d period was good (67 and 64% for PCR and culture results, respectively), which justifies the utility of environmental sampling to identify pens with a high MAP bioburden between routine cow pen changes on a dairy. Environmental sampling of free-stall pens using the standardized sampling protocol yielded comparable PCR and culture results across collectors with different experience levels and at different times within a 5-d period.

High magnetic relaxivity in a fluorescent CdSe/CdS/ZnS quantum dot functionalized with MRI contrast molecules.

Light emitting semiconducting quantum dots show great promise as solar cells, optoelectronic devices and multimodal imaging probes. Here we demonstrate successful grafting of a thiol-functionalised GdIII MRI contrast agent onto the surface of core-multishell CdSe/CdS/ZnS quantum dots. The resulting nanoprobe exhibits intense photoluminescence and unprecedentedly large T1 relaxivity of 6800 mM-1 s-1 per nanoparticle due to secure implanting of ca. 620 magnetic centers per quantum dot unit.

Medium hyposmolarity stimulates prolactin secretion in GH4C1 cells by inducing an increase in cytosolic free calcium.

Extracellular hyposmolarity is a potent direct stimulus for hormone secretion for which a mechanism has not been delineated. The importance of plasmalemma Ca2+ permeability in this phenomenon in pituitary tumor-derived GH4C1 cells was evaluated by comparing the dynamics of changes in cytosolic free Ca2+ concentration [( Ca2+]i) with those of PRL secretion. At a normal physiological concentration of extracellular Ca2+ (1.5 mM), hyposmolarity induced a striking rise in both [Ca2+]i and PRL secretion, which was proportional to the stimulus between 0.50% reduction in medium osmolarity. Thirty percent hyposmolarity induced a 3-fold rise in [Ca2+]i and a 5-fold rise in PRL secretion above the basal level. These effects did not occur in cells incubated in a medium with a Ca2+ concentration lower than 30 microM. In cells incubated in 1.5 mM Ca2+, the Ca2(+)-channel antagonists, nifedipine and verapamil, significantly inhibited hyposmolar-induced increases in [Ca2+]i and PRL secretion. These data suggest that in GH4C1 cells medium hyposmolarity causes a burst of PRL secretion that depends on a similar preceding rise in [Ca2+]i produced by extracellular Ca2+ influx, most of which passes through dihydropyridine-sensitive Ca2(+)-channels.

Evidence that ethanol induces prolactin secretion in GH4C1 cells by producing cell swelling with resultant calcium influx.

Although acute exposure to ethanol has been reported to affect hormone secretion, the data are sometimes conflicting, and the mechanism of action of ethanol is unclear. We have examined in GH4C1 cells the effect of isotonic ethanol on cell volume measured with a Coulter counter, the cytosolic Ca2+ concentration ([Ca2+]i) monitored with fura-2, and PRL secretion analyzed in a perifusion system. Isotonic ethanol caused prompt cell swelling and an explosive rise in both [Ca2+]i and PRL secretion proportional to the concentration of ethanol between 5-120 mM. The increases in both [Ca2+]i and PRL secretion induced by 80 mM isotonic ethanol were essentially abolished by removal of medium Ca2+ or by nifedipine; the nifedipine IC50 was approximately 20 nM. Cell swelling induced by hyposmolarity or isotonic urea similarly increased both [Ca2+]i and PRL secretion. Hypertonic ethanol did not cause cell swelling and was ineffective in inducing an increase in either [Ca2+]i or PRL secretion. These data suggest that in GH4C1 cells a major mechanism by which ethanol stimulates PRL secretion is to induce cell swelling, thus producing enhanced Ca2+ influx through dihydropyridine-sensitive Ca2+ channels.

The permeant molecule urea stimulates prolactin secretion in GH4C1 cells by inducing Ca2+ influx through dihydropyridine-sensitive Ca2+ channels.

Isotonic urea in medium with a normal 1.2 mM Ca2+ concentration induced a striking rise in both cytosolic Ca2+ concentration ([Ca2+]i) and prolactin (PRL) secretion, each of whose peaks were proportional to the concentration of urea between 5 and 120 mM. There was a significant linear relationship between the peaks of induced [Ca2+]i and PRL secretion (r = 0.99, P less than 0.001). The increase in both [Ca2+]i and PRL secretion was completely abolished by removal of medium Ca2+ or by 2 microM nifedipine. Hypertonic urea was ineffective in inducing either an increase in [Ca2+]i or PRL secretion. These data support the hypothesis that plasma membrane expansion is a potent non-toxic inducer of hormone secretion and that in GH4C1 cells an increase in [Ca2+]i produced by enhanced influx of extracellular Ca2+ through dihydropyridine-sensitive Ca2+ channels plays an important role in this phenomenon.

Similarity, invariance, and musical variation.

Perceptual similarity underlies a number of important psychological properties of musical materials, including perceptual invariance under transformation, categorization, recognition, and the sense of familiarity. Mental processes involved in the perception of musical similarity may be an integral part of the functional logic of music composition and thus underly important aspects of musical experience. How much and in what ways can musical materials be varied and still be considered as perceptually related or as belonging to the same category? The notions of musical material, musical variation, perceptual similarity and invariance, and form-bearing dimensions are considered in this light. Recent work on similarity perception has demonstrated that the transformation space for a given musical material is limited by several factors ranging from degree of match of the values of auditory attributes of the events composing the sequences to their relations of various levels of abstraction and to the degree that the transformation respects the grammar of the musical system within which the material was composed. These notions and results are considered in the light of future directions of research, particularly concerning the role of similarity and invariance in the understanding of musical form during listening.

Isolation, genetic diversity, and geographic distribution of Bayou virus (Bunyaviridae: hantavirus).

Bayou hantavirus, previously implicated in human hantavirus pulmonary syndrome in Louisiana, was isolated from a rice rat (Oryzomys palustris) captured in Georgia. The presence of antibody among rice rats captured throughout the southeastern United States and the extent of diversity among the genetic variants of Bayou viruses suggest that the rice rat is the most likely natural reservoir of the virus and that both virus and host have probably co-evolved for some years.

Comparison of isoproterenol and dibutyryl adenosine cyclic 3',5'-monophosphate stimulation of thyroxine 5'-deiodinase activity in cultured pineal glands from euthyroid and hypothyroid rats.

Thyroxine 5'-deiodinase was increased by isoproterenol and dibutyryl adenosine cyclic 3',5'-monophosphate in a dose- and time-related manner in cultured rat pineal gland. Basal and stimulated activity was higher in glands from hypothyroid than from euthyroid animals. Our data suggest direct beta-adrenergic stimulation of intracellular cyclic AMP may be involved in the regulation of pineal thyroxine 5'-deiodinase activity.

Dual effect of osmotic cell swelling on prolactin secretion by acutely dispersed adenohypophyseal cells.

Cell swelling induced by acute exposure to the permeant molecule urea or by medium hyposmolarity evoked a prompt PRL secretory burst from dispersed rat anterior pituitary cells. However, during continuous exposure greater than or equal to 10 min to these conditions inhibition of basal and TRH-induced PRL secretion occurred and there was an "off" burst of PRL secretion following return to basal conditions. Compared with continuous TRH stimulation which causes biphasic PRL secretion with a rapid high amplitude first phase secretory burst followed by a sustained low level second phase of secretion, cell swelling induced only "first phase" secretion. Removing Ca2+ from the medium or adding 50 microM verapamil markedly depressed the "off" secretory burst following return to basal conditions but had no effect on the initial high amplitude burst. Our data suggest that the effect of cell swelling on PRL secretion is complex and that there are at least two mechanisms for PRL secretion in normal anterior pituitary cells; these are differently affected by cell swelling and Ca2+ influx.

Dichotic perception and laterality in neonates.

Groups of 4-day-old neonates were tested for dichotic discrimination and ear differences with the High-Amplitude-Sucking procedure. In the first experiment, dichotic speech discrimination was attested by comparison with a control group. Furthermore, among those subjects who showed a substantial recovery of sucking response at least after one of the two syllable changes, it was observed that significantly more subjects manifested a stronger reaction to a right-ear change than to a left-ear change. In the second experiment, 4-day-old neonates were tested on syllable and music timbre discrimination. The significant Stimulus Type x Ear interaction observed suggests perceptual asymmetries indicative of very precocious brain specialization.

Prevalence of Mycobacterium avium subsp. paratuberculosis fecal shedding in alpacas presented to veterinary hospitals in the United States.

BACKGROUND: The prevalence of Johne's disease in alpacas in the United States is unknown. The limits of polymerase chain reaction (PCR) detection of Mycobacterium avium subsp. paratuberculosis (MAP) in alpaca feces have not been determined. OBJECTIVES: To evaluate the use of PCR for MAP detection in alpaca feces; and to estimate the prevalence of MAP fecal shedding in alpacas presented to veterinary teaching hospitals. ANIMALS: Alpacas presenting to 4 US veterinary teaching hospitals from November 2009 to February 2011. METHODS: Prospective study. Ten dilutions of a wild MAP strain were added to negative alpaca feces and processed for MAP detection by means of a commercial real-time PCR (RT-PCR) assay, and cultured on Herrold's Egg Yolk Medium (HEYM) and liquid broth. The limits of detection for each method were determined. Fecal samples from alpacas admitted to the veterinary teaching hospitals during the study period were evaluated for MAP via PCR and HEYM. RESULTS: The lowest MAP dilution detectable via PCR was 243 MAP colony-forming units (CFU)/g of feces, at which concentration MAP growth was detectable on HEYM. Ten (6%; 95% confidence interval: 3-9%) of the 180 fecal samples collected were positive on PCR. CONCLUSIONS AND CLINICAL IMPORTANCE: Polymerase chain reaction can provide an accurate and rapid detection of MAP fecal shedding in alpacas; and the prevalence of MAP fecal shedding in hospitalized alpacas in 4 US veterinary teaching hospitals was 6%.

Antimicrobial activity of gallium nitrate against Mycobacterium avium subsp. paratuberculosis in neonatal calves.

BACKGROUND: Mycobacterium avium subsp. paratuberculosis (MAP), the agent of Johne's disease in cattle, is a facultative intracellular bacterium that is dependent on ferric iron for its survival and replication. Gallium (Ga), a trivalent semimetal that shares many similarities with ferric iron and functions as an iron mimic has been shown to have in vitro antimicrobial activity against several microorganisms, including MAP. OBJECTIVES: (1) To investigate the antimicrobial activity of Ga in calves experimentally infected with MAP; and (2) to monitor for potential adverse effects of Ga on calf health. ANIMALS: Twelve Holstein calves. METHODS: Randomized blind controlled experiment. Beginning at 10 days of age (study day 1), the experimental calves (n = 6) were treated with 20 mg/kg gallium nitrate daily for 45 days. On study days 4 and 5, all calves were challenged with a PO dose of a live field strain MAP. Treated calves were monitored daily for adverse effects. Calves were euthanized on study day 100, and 29 tissue samples and 1 fecal sample were collected from each calf. Samples were cultured for MAP by MGIT liquid culture system, Herrold's Egg Yolk Medium culture, or both. RESULTS: No adverse effects were observed in the treated calves. Treatment was associated with a significant reduction in MAP tissue burden when compared with control calves (P = .017). CONCLUSIONS AND CLINICAL RELEVANCE: Chemoprophylactic treatment of calves with Ga before and during the period of high susceptibility decreased MAP tissue colonization in experimentally infected neonatal calves.

Segregation of concurrent sounds. I: Effects of frequency modulation coherence.

Frequency modulation coherence was investigated as a possible cue for the perceptual segregation of concurrent sound sources. Synthesized chords of 2-s duration and comprising six permutations of three sung vowels (/a/, /i/, /o/) at three fundamental frequencies (130.8, 174.6, and 233.1 Hz) were constructed. In one condition, no vowels were modulated, and, in a second, all three were modulated coherently such that the ratio relations among all frequency components were maintained. In a third group of conditions, one vowel was modulated, while the other two remained steady. In a fourth group, one vowel was modulated independently of the two other vowels, which were modulated coherently with one another. Subjects were asked to judge the perceived prominence of each of the three vowels in each chord. Judged prominence increased significantly when the target vowel was modulated compared to when it was not, with the greatest increase being found for higher fundamental frequencies. The increase in prominence with modulation was unaffected by whether the target was modulated coherently or not with nontarget vowels. The modulation and pitch position of nontarget vowels had no effect on target vowel prominence. These results are discussed in terms of possible concurrent auditory grouping principles.

Two phase effects in roughness perception.

The respective influences of spectral and temporal aspects of sound in roughness perception are examined by way of phase manipulations. In a first experiment, the phase of the central component of three-component signals is shown to modify perceived roughness, for a given amplitude spectrum, regardless of whether it modifies the waveform envelope. A second experiment shows that the shape of the waveform envelope, for a given amplitude spectrum and a given modulation depth, also influences perceived roughness. We interpret both of these results by considering the envelope of an internal representation that is deduced from the physical signal by taking into account peripheral auditory processing. The results indicate that the modulation depth of such an internal representation is not the only determinant of roughness, but that an effect of temporal asymmetry is also to be taken into account.

Organization and discrimination of repeating sound sequences by newborn infants.

A study was conducted to determine whether newborn infants organize auditory streams in a manner similar to that of adults. A series of three experiments investigated the ability of 3- to 4-day-old infants to discriminate repeated rising and falling four-tone sequences in two configurations of source timbre and spatial position. It was hypothesized that if the sequences were organized into two auditory streams on the basis of timbre and spatial position, one of the configurations should be discriminable from its reversal while the other should not. The sequences were tested with different pitch and temporal intervals separating the tones. Sequences were discriminated for the first configuration by adults at both fast tempo/small interval and slow tempo/large interval combinations, while only the latter was discriminated by newborns as measured with a non-nutritive high-amplitude sucking paradigm. Neither adults nor infants could discriminate the sequence reversals for the second configuration. The results suggest that newborn infants organize auditory streams on the basis of source timbre and/or spatial position. They also suggest that newborns have limits in temporal and/or pitch resolution when discriminating tone sequences.

The auditory continuity phenomenon: role of temporal sequence structure.

The auditory continuity phenomenon occurs when listening to a series of alternating high- and low-level tones: instead of perceiving this intermittence, listeners often report hearing a continuous tone upon which is superimposed a series of intermittent tones. The temporal limits to the perception of this phenomenon are investigated, as well as effects of task instructions on it. A loudness-matching paradigm developed previously [McAdams et al., J. Acoust. Soc. Am. 130, 1580-1591 (1998)] provided both an objective indication of this phenomenon and an indication of its strength. In the studies reported here, the phenomenon was observed reliably when the low-level tones were at least half the duration of the high-level tones: the greater the duration ratio between low-level and high-level tones, the stronger the phenomenon. Duty-cycle duration did not affect the strength of this phenomenon. It proved to be particularly robust as its strength was unaffected by task instructions, level of expertise, and repetition. A model is proposed in which adjusted levels depend on the relative levels of the high- and low-level tones weighted by relative duration and attentional focusing.

Simple selected ion monitoring capillary gas chromatographic-mass spectrometric method for the determination of cotinine in serum, urine and oral samples.

A capillary gas chromatographic-mass spectrometric method using selected ion monitoring was developed for the analysis of cotinine in urine, serum and oral samples. The procedure requires 500 microliters of an oral sample, 250 microliters of a serum sample and 50 microliters of urine and can detect 5 ng/ml cotinine in oral samples, 10 ng/ml in serum and 50 ng/ml in urine with good precision and accuracy. The method was used to determine the cotinine concentration in samples of all three fluids collected from a group of smokers and non-smokers.

Segregation of concurrent sounds. II: Effects of spectral envelope tracing, frequency modulation coherence, and frequency modulation width.

The research presented here concerns the simultaneous grouping of the components of a vocal sound source. McAdams [J. Acoust. Soc. Am. 86, 2148-2159 (1989)] found that when three simultaneous vowels at different pitches were presented with subaudio frequency modulation, subjects judged them as being more prominent than when no vibrato was present. In a normal voice, when the harmonics of a vowel undergo frequency modulation they also undergo an amplitude modulation that traces the spectral envelope. Hypothetically, this spectral tracing could be one of the criteria used by the ear to group components of each vowel, which may help explain the lack of effect of frequency modulation coherence among different vowels in the previous study. In this experiment, two types of vowel synthesis were used in which the component amplitudes of each vowel either remained constant with frequency modulation or traced the spectral envelope. The stimuli for the experiment were chords of three different vowels at pitch intervals of five semitones (ratio 1.33). All the vowels of a given stimulus were produced by the same synthesis method. The subjects' task involved rating the prominence of each vowel in the stimulus. It was assumed that subjects would judge this prominence to be lower when they were not able to distinguish the vowel from the background sound. Also included as stimulus parameters were the different permutations of the three vowels at three pitches and a number of modulation conditions in which vowels were unmodulated, modulated alone, and modulated either coherently with, or independently of, the other vowels. Spectral tracing did not result in increased ratings of vowel prominence compared to stimuli where no spectral tracing was present. It would therefore seem that it has no effect on grouping components of sound sources. Modulated vowels received higher prominence ratings than unmodulated vowels. Vowels modulated alone were judged to be more prominent than vowels modulated with other vowels. There was, however, no significant difference between coherent and independent modulation of the three vowels. Differences among modulation conditions were more marked when the modulation width was 6% than when it was 3%.

Unusual manifestations of peripartal cardiac disease.

Because peripartal cardiac disease occurs infrequently, its manifestations may be unfamiliar to most physicians. We report two unusual cases of postpartum cardiac disease: one patient presented with cerebral and peripheral arterial embolization, and the second patient developed late eclamptic seizures with subsequent myocardial infarction. Both patients recovered. Nonobstetric physicians should be aware of these pregnancy-associated medical complications to allow prompt diagnosis and aggressive therapy.