Measurement of the parity-violating asymmetry in inclusive electroproduction of π- near the Δ0 resonance.

The parity-violating (PV) asymmetry of inclusive π- production in electron scattering from a liquid deuterium target was measured at backward angles. The measurement was conducted as a part of the G0 experiment, at a beam energy of 360 MeV. The physics process dominating pion production for these kinematics is quasifree photoproduction off the neutron via the Δ0 resonance. In the context of heavy-baryon chiral perturbation theory, this asymmetry is related to a low-energy constant d(Δ)- that characterizes the parity-violating γNΔ coupling. Zhu et al. calculated d(Δ)- in a model benchmarked by the large asymmetries seen in hyperon weak radiative decays, and predicted potentially large asymmetries for this process, ranging from A(γ)-=-5.2 to +5.2 ppm. The measurement performed in this work leads to A(γ)-=-0.36±1.06±0.37±0.03 ppm (where sources of statistical, systematic and theoretical uncertainties are included), which would disfavor enchancements considered by Zhu et al. proportional to V(ud)/V(us). The measurement is part of a program of inelastic scattering measurements that were conducted by the G0 experiment, seeking to determine the N-Δ axial transition form factors using PV electron scattering.

Observation of two-neutrino double-beta decay in 136Xe with the EXO-200 detector.

We report the observation of two-neutrino double-beta decay in (136)Xe with T(1/2) = 2.11 ± 0.04(stat) ± 0.21(syst) × 10(21) yr. This second-order process, predicted by the standard model, has been observed for several nuclei but not for (136)Xe. The observed decay rate provides new input to matrix element calculations and to the search for the more interesting neutrinoless double-beta decay, the most sensitive probe for the existence of Majorana particles and the measurement of the neutrino mass scale.

Transverse beam spin asymmetries at backward angles in elastic electron-proton and quasielastic electron-deuteron scattering.

We have measured the beam-normal single-spin asymmetries in elastic scattering of transversely polarized electrons from the proton, and performed the first measurement in quasielastic scattering on the deuteron, at backward angles (lab scattering angle of 108°) for Q² = 0.22 GeV²/c² and 0.63 GeV²/c² at beam energies of 362 and 687 MeV, respectively. The asymmetry arises due to the imaginary part of the interference of the two-photon exchange amplitude with that of single-photon exchange. Results for the proton are consistent with a model calculation which includes inelastic intermediate hadronic (πN) states. An estimate of the beam-normal single-spin asymmetry for the scattering from the neutron is made using a quasistatic deuterium approximation, and is also in agreement with theory.

The pre-analytical challenges of routine urinalysis.

An effective diagnostic strategy for urinalysis should be based on standard procedures for collection, transport, sample preparation and analysis. In view of a better reproducibility of the analyses, the pre-analytical requirements become stricter. Various sample methods can cause significant pre-analytical errors. It is a challenge for the laboratory to control the steps in the pre-analytical phase that contribute to pre-analytical variability. To reduce the variability, it is necessary to look at the pre-analytical process as a complete entity, from test ordering to the moment of specimen processing. Clinical laboratories are responsible for the clinical and financial outcome of this phase. In a culture of increasing productivity, lower costs and improving quality, the challenge is to use several tools designed to standardize and optimize urinalysis. Despite advances in the performance of analytic systems, the pre-analytical phase of modern urinalyses has not been studied very thoroughly. This review of the literature lights on different problems in current pre-analytical requirements for particle and test strip analysis of urine samples.

Strange quark contributions to parity-violating asymmetries in the backward angle G0 electron scattering experiment.

We have measured parity-violating asymmetries in elastic electron-proton and quasielastic electron-deuteron scattering at Q2=0.22 and 0.63 GeV2. They are sensitive to strange quark contributions to currents in the nucleon and the nucleon axial-vector current. The results indicate strange quark contributions of approximately < 10% of the charge and magnetic nucleon form factors at these four-momentum transfers. We also present the first measurement of anapole moment effects in the axial-vector current at these four-momentum transfers.

Detection of antigenic heterogeneity in feline coronavirus nucleocapsid in feline pyogranulomatous meningoencephalitis.

A new monoclonal antibody (mAb), CCV2-2, was compared with the widely used FIPV3-70 mAb, both directed against canine coronavirus (CCoV), as a diagnostic and research tool. Western blot showed that both anti-CCoV mAbs only reacted with a protein of 50 kD, a weight consistent with the feline coronavirus (FCoV) viral nucleocapsid. A competitive inhibition enzyme-linked immunosorbent assay showed that the 2 recognized epitopes are distinct. Preincubation of CCV2-2 mAb with FCoV antigen suppressed the immunostaining. Formalin-fixed, paraffin-embedded sections from brains of 15 cats with the dry form of feline infectious peritonitis (FIP) were examined by immunohistochemistry. Immunohistochemistry was performed with both anti-CCoV mAbs, either on consecutive or on the same sections. A myeloid-histiocytic marker, MAC 387, was also used to identify FIP virus-infected cells. In all regions where MAC 387-positive cells were present, positive staining with the CCV2-2 mAb was systematically detected, except at some levels in 1 cat. In contrast, none or only a few cells were positive for the FIPV3-70 mAb. Double immunostaining showed macrophages that were immunopositive for either CCV2-2 alone or alternatively for CCV2-2 and FIPV3-70 mAbs. This reveals the coexistence of 2 cohorts of phagocytes whose FIP viral contents differed by the presence or absence of the FIPV3-70-recognized epitope. These findings provide evidence for antigenic heterogeneity in coronavirus nucleocapsid protein in FIP lesions, a result that is in line with molecular observations. In addition, we provide for the first time morphologic depiction of viral variants distribution in these lesions.

Naturally occurring parvovirus-associated feline hypogranular cerebellar hypoplasia-- A comparison to experimentally-induced lesions using immunohistology.

Three cases of feline cerebellar hypoplasia are presented. At the time of examination, the ages of the cats ranged from 2 months to 1 year. Necropsy revealed cerebellar and pons hypoplasia. Polymerase chain reaction for parvoviral deoxyribonucleic acid was positive in cerebellar tissue. Cell-specific immunolabeling was used to characterize the lesions, which were characterized into 2 types. In type 1 lesions, the cortex was nearly agranular, with an extremely thin molecular layer; the Purkinje cells were randomly placed and oriented, and their stunted main dendrite produced a thorn-covered atrophic dendritic tree; the basket cell axons ran randomly and had dysmorphic endings; and myelinated fibers were severely reduced in folia axes. In type 2 lesions, the cortex was hypogranular; the Purkinje cells were linearly organized, but their main dendrite extended too far in the molecular layer before giving up smooth, bent secondary dendrites; many basket cells were located along the cerebellar surface, and their axons ran at right angle to the surface; myelinated fibers were moderately reduced. Defects in climbing fiber synapse translocation and elimination were evident in both types of lesion. This immunohistologic study allowed a comparison between lesions in these spontaneous cerebellar hypoplasia cases with those documented when using silver impregnation studies after perinatal experimental cerebellar damage. Such a comparison is consistent with viral infection that occurs before birth in all 3 cases. Progress in parvovirus biology knowledge suggests that viral NS1 protein cytotoxicity might explain degenerative changes in the Purkinje cells that were present, in addition to the development defect.

Implementation of a process analytical technology system in a freeze-drying process using Raman spectroscopy for in-line process monitoring.

The aim of the present study was to propose a strategy for the implementation of a Process Analytical Technology system in freeze-drying processes. Mannitol solutions, some of them supplied with NaCl, were used as models to freeze-dry. Noninvasive and in-line Raman measurements were continuously performed during lyophilization of the solutions to monitor real time the mannitol solid state, the end points of the different process steps (freezing, primary drying, secondary drying), and physical phenomena occurring during the process. At-line near-infrared (NIR) and X-ray powder diffractometry (XRPD) measurements were done to confirm the Raman conclusions and to find out additional information. The collected spectra during the processes were analyzed using principal component analysis and multivariate curve resolution. A two-level full factorial design was used to study the significant influence of process (freezing rate) and formulation variables (concentration of mannitol, concentration of NaCl, volume of freeze-dried sample) upon freeze-drying. Raman spectroscopy was able to monitor (i) the mannitol solid state (amorphous, alpha, beta, delta, and hemihydrate), (ii) several process step end points (end of mannitol crystallization during freezing, primary drying), and (iii) physical phenomena occurring during freeze-drying (onset of ice nucleation, onset of mannitol crystallization during the freezing step, onset of ice sublimation). NIR proved to be a more sensitive tool to monitor sublimation than Raman spectroscopy, while XRPD helped to unravel the mannitol hemihydrate in the samples. The experimental design results showed that several process and formulation variables significantly influence different aspects of lyophilization and that both are interrelated. Raman spectroscopy (in-line) and NIR spectroscopy and XRPD (at-line) not only allowed the real-time monitoring of mannitol freeze-drying processes but also helped (in combination with experimental design) us to understand the process.

Brain stem auditory potentials evoked by clicks in the presence of high-pass filtered noise in dogs.

This study evaluates the effects of a high-frequency hearing loss simulated by the high-pass-noise masking method, on the click-evoked brain stem-evoked potentials (BAEP) characteristics in dogs. BAEP were obtained in response to rarefaction and condensation click stimuli from 60 dB normal hearing level (NHL, corresponding to 89 dB sound pressure level) to wave V threshold, using steps of 5 dB in eleven 58 to 80-day-old Beagle puppies. Responses were added, providing an equivalent to alternate polarity clicks, and subtracted, providing the rarefaction-condensation potential (RCDP). The procedure was repeated while constant level, high-pass filtered (HPF) noise was superposed to the click. Cut-off frequencies of the successively used filters were 8, 4, 2 and 1 kHz. For each condition, wave V and RCDP thresholds, and slope of the wave V latency-intensity curve (LIC) were collected. The intensity range at which RCDP could not be recorded (pre-RCDP range) was calculated. Compared with the no noise condition, the pre-RCDP range significantly diminished and the wave V threshold significantly increased when the superposed HPF noise reached the 4 kHz area. Wave V LIC slope became significantly steeper with the 2 kHz HPF noise. In this non-invasive model of high-frequency hearing loss, impaired hearing of frequencies from 8 kHz and above escaped detection through click BAEP study in dogs. Frequencies above 13 kHz were however not specifically addressed in this study.

Dark-cell areas in the dog vestibular endorgans: an immunohistochemical study.

The stria vascularis in the cochlea and the dark-cell areas in the vestibular endorgans are structures involved in the production of endolymphatic fluid. This study investigated the dark-cell areas in the vestibular endorgans of the dog by classical staining and by immunohistochemistry (anti-Na,K-ATPase beta2 isoform, anti-cytokeratins (against cytokeratins 5 and 8), anti-vimentin and anti-S100A6) from birth to 110 postnatal days. Using classical staining, it was not possible to discriminate dark cells from other epithelial cells lining the vestibular endolymphatic spaces. From birth, the Na,K-ATPase beta2 isoform was expressed in the lateral and basal cell membranes of a subset of cells located in the utricular wall, at the base of the cristae ampullaris and was identified as dark cells. From birth, anti-cytokeratins labelled all the cells forming the epithelial lining, including the dark cells, while anti-vimentin labelled the underlying mesenchymal cells. From postnatal day 10, anti-S100A6 labelled subepithelial cells exclusively located underneath the dark-cell areas and were identified as vestibular melanocyte-like cells. From birth, Fontana staining evidenced fine melanin granules in the subepithelial layer. The amount of melanin granules increased during the first month. Melanin distribution was closely associated with the region where S100A6-positive cells were located. The cell-specific antigen expression in the dog dark-cell areas was clearly comparable to that of the dog stria vascularis previously described. The present investigation also suggested an earlier histological and immunohistological maturity in the dark-cell areas than in the stria vascularis of dogs. This preliminary morphological description of the normal dark-cell areas in dogs by means of immunomarkers may be instrumental in studying pathological processes involving the fluid-secreting structures in vestibular endorgans.

Inner ear morphology in a bilaterally deaf Dogo Argentino pup.

Two bilaterally deaf and three unilaterally deaf pups were identified from a litter of 10 Dogo Argentino pups presented for hearing evaluation by electrophysiological investigation. One pup, a bilaterally deaf female aged 43 days, was available for histopathology. Examination of both inner ears revealed bilateral cochlear degeneration with atrophy of the stria vascularis, collapse of the cochlear duct, degeneration of the organ of Corti, and abnormal tectorial membrane. The left vestibule, including the sacculus, was normal. The spiral and vestibular ganglia were essentially normal. This is the first histopathological description of lesions associated with deafness in a Dogo Argentino, but abnormalities were similar to those previously described in deaf Dalmatian pups and in other white hair-coated breeds. The defect was classified as a cochleosaccular degeneration. It was probably congenital and genetic causes were suspected.

An original inner ear neuroepithelial degeneration in a deaf Rottweiler puppy.

Histopathological investigation was conducted on both inner ears from a 4.5-month-old Rottweiler puppy with electrophysiologically confirmed bilateral deafness. The lesions were restricted to the organ of Corti and spiral ganglion that both displayed severe degenerative changes. The outer hair cells were less affected than the inner hair cells. The number of spiral ganglion neurons was reduced, and remaining neurons were altered. The basal and middle cochlear turns were more affected than the apical one. The vestibules were normal. Immunostaining with calbindin, calretinin, S100A1 and S100A6 polyclonal antisera was helpful in identifying different cell-types in the degenerated cochlea. The early and severe spiral ganglion cell degeneration is an uncommon finding no matter the species. Such lesions bear significance within the frame of cochlear implants technology for deaf infants.

Immunolocalization of the calcium binding S100A1, S100A5 and S100A6 proteins in the dog cochlea during postnatal development.

The immunolocalization of three members of the S100 calcium-binding protein family was investigated in the dog cochlea during normal postnatal development. Sections of decalcified and paraffin-embedded cochleae from 16 beagle puppies aged from birth to 3 months were treated with polyclonal antisera raised against the human recombinant S100A1, S100A5, and S100A6 proteins. At birth, in the dog cochlea, S100A1 was expressed in the immature Deiter's cells, and slightly in the pillar cells. From the second week, S100A1 was detected in the supporting structures of the organ of Corti, i.e. the Deiter's, the pillar, the border, and the Hensen's cells, and in the reticular membrane. From birth onwards, S100A5 remained a neuronal-specific protein, only located in a subpopulation of neurons in the spiral ganglion. S100A6 was not expressed at birth. From the second week of life, the Schwann cells and nerve sheaths in the modiolus, in the spiral ganglion, and running in the direction of the organ of Corti exhibited S100A6-labeling. From the 12th postnatal day, some scattered intermediate cells started to express S100A6 protein in the stria vascularis. The number of labeled intermediate cells increased during the third week. At adult stage, the intermediate cells were S100A6-stained with cytoplasmic labeling throughout the stria vascularis from the base to the apex of the cochlea. None of the other cochlear structures expressed the S100 proteins under study during the postnatal development of the dog cochlea. The S100A1, S100A5, S100A6 immunostaining was limited to specific cell types in dog cochlea. These S100 proteins were useful markers in the study of supporting cells, neurons, nerve fibers sheaths and stria vascularis (S100A6) during the normal postnatal development of the dog cochlea.

Decreased benzodiazepine receptor density in the cerebellum of early blind human subjects.

As a first approach to study the effect of early visual deprivation in the GABA-ergic inhibitory system, the distribution of benzodiazepine receptors (BZR) was accurately estimated using [11C]flumazenil ([11C]FMZ). Measurements were carried out in five subjects who became blind early in life and in five sighted control subjects. The interactions between [11C]FMZ and BZR were described using a non-linear compartmental analysis which permitted to estimate the BZR synaptic density independently of other model parameters. The distribution of BZR in the visual areas and other cortical regions of blind subjects was qualitatively and quantitatively similar to that of controls. However, the BZR density in the cerebellum was significantly lower in blind than in control subjects (P<0.01). Our findings suggest that modifications of the cerebellar neural circuitry may be concomitant to the already observed compensatory reorganization in cerebral areas of blind subjects.

Maturation of the auditory system in clinically normal puppies as reflected by the brain stem auditory-evoked potential wave V latency-intensity curve and rarefaction-condensation differential potentials.

OBJECTIVE: To evaluate auditory maturation in puppies. ANIMALS: Ten clinically normal Beagle puppies. PROCEDURE: Puppies were examined repeatedly from days 11 to 36 after birth (8 measurements). Click-evoked brain stem auditory-evoked potentials (BAEP) were obtained in response to rarefaction and condensation click stimuli from 90 dB normal hearing level to wave V threshold, using steps of 10 dB. Responses were added, providing an equivalent to alternate polarity clicks, and subtracted, providing the rarefaction-condensation differential potential (RCDP). Steps of 5 dB were used to determine thresholds of RCDP and wave V. Slope of the low-intensity segment of the wave V latency-intensity curve was calculated. The intensity range at which RCDP could not be recorded (ie, pre-RCDP range) was calculated by subtracting the threshold of wave V from threshold of RCDP RESULTS: Slope of the wave V latency-intensity curve low-intensity segment evolved with age, changing from (mean +/- SD) -90.8 +/- 41.6 to -27.8 +/- 4.1 micros/dB. Similar results were obtained from days 23 through 36. The pre-RCDP range diminished as puppies became older, decreasing from 40.0 +/- 7.5 to 20.5 +/- 6.4 dB. CONCLUSION AND CLINICAL RELEVANCE: Changes in slope of the latency-intensity curve with age suggest enlargement of the audible range of frequencies toward high frequencies up to the third week after birth. Decrease in the pre-RCDP range may indicate an increase of the audible range of frequencies toward low frequencies. Age-related reference values will assist clinicians in detecting hearing loss in puppies.

Comparison of regional cerebral blood flow and glucose metabolism in the normal brain: effect of aging.

The regional cerebral blood flow (rCBF) and metabolic rate for glucose (rCMRGlc) are associated with functional activity of the neural cells. The present work reports a comparison study between rCBF and rCMRGlc in a normal population as a function of age. 10 young (25.9+/-5.6 years) and 10 old (65.4+/-6.1 years) volunteers were similarly studied at rest. In each subject, rCBF and rCMRGlc were measured in sequence, during the same session. Both rCBF and rCMRGlc values were found to decrease from young (mean rCBF=43.7 ml/100 g per min; mean rCMRGlc=40.6 micromol/100 g per min) to old age (mean rCBF=37.3 ml/100 g per min; mean rCMRGlc=35.2 micromol/100 g per min), resulting in a drop over 40 years of 14.8% (0.37%/year) and 13.3% (0.34%/year), respectively. On a regional basis, the frontal and the visual cortices were observed to have, respectively, the highest and the lowest reduction in rCBF, while, for rCMRGlc, these extremes were observed in striatum and cerebellum. Despite these differences, the ratio of rCBF to rCMRGlc was found to have a similar behavior in all brain regions for young and old subjects as shown by a correlation coefficient of 88%. This comparative study indicates a decline in rCBF and rCMRGlc values and a coupling between CBF and CMRGlc as a function of age.

Myeloid leukemic cells express and secrete bioactive pituitary-sized 23 kDa prolactin.

Prolactin (PRL) is a 23 kDa polypeptide hormone of pituitary origin which is of major importance for reproduction. In addition, PRL has immunomodulatory effects and can be produced in small quantities in nonpituitary tissues. To address possible autocrine or paracrine functions of PRL in leukemia, we characterized immunoreactive PRL from the culture medium of leukemic cells. The myeloid cell line Eol-1 expresses the long extrapituitary type mRNA for PRL and synthesizes immunoreactive PRL with a molecular weight of 23 kDa. The biological activity in Eol-1 culture medium was determined using the Nb2 bioassay. This activity co-eluted with recombinant human (rh) PRL on an S-200 Sephacryl gel filtration column and could be blocked by anti-PRL antiserum. Western blot analysis and Nb2 bioassays also suggest that acute myelogenous leukemic blasts secrete bioactive 23 kDa PRL in one out of three tested patients.

Brainstem auditory evoked potential wave V latency-intensity function in normal Dalmatian and Beagle puppies.

This study investigated whether Dalmatian puppies with normal hearing bilaterally had the same click-evoked brainstem auditory potential characteristics as age-matched dogs of another breed. Short-latency brainstem auditory potentials evoked by condensation and rarefaction clicks were recorded in 23 1.5- to 2-month-old Dalmatian puppies with normal hearing bilaterally by a qualitative brainstem auditory evoked potential test and in 16 Beagle dogs of the same age. For each stimulus intensity, from 90 dB normal hearing level down to the wave V threshold, the sum of the potentials evoked by the 2 kinds of stimuli were added, giving an equivalent to the alternate click polarity stimulation. The slope of the L segment of the wave V latency-intensity curve was steeper in Dalmatian (-40 +/- 10 micros/dB) than in Beagles (-28 +/- 5 micros/dB, P < .001) puppies. The hearing threshold was lower in the Beagle puppies (P < .05). These results suggest that interbreed differences may exist at the level of cochlear function in this age class. The wave V latency and wave V-wave I latencies differences at high stimulus intensity were different between the groups of puppies (4.3 +/- 0.2 and 2.5 +/- 0.2 milliseconds, respectively, for Beagles; and 4.1 +/- 0.2 and 2.3 +/- 0.2 milliseconds for Dalmatians, P < .05). A different maturation speed of the neural pathways is one possible explanation of this observation.

Immunolocalization of calbindin D28k and calretinin in the dog cochlea during postnatal development.

The calbindin (CB) and the calretinin (CR) immunoreactivities were studied in the dog cochlea during its postnatal maturation from birth to the 33rd postnatal day. At birth, CB was expressed in the Kölliker's organ, in the immature inner (IHC) and outer hair cells (OHC), in neurons of the spiral ganglion, and in nerve fibers running in the basilar membrane of the apical turn. During the cochlear maturation, non-sensorineuronal structures, such as the Kölliker's organ, the rods of Corti, and the inner sulcus cells, displayed a transient CB-staining. In the adult-like dog cochlea, CB was found in the cytoplasm, the cuticular plate, and the stereocilia of the IHC and OHC. All the neurons of the spiral ganglion and some nerves fibers in the modulius were CB-positive. At birth, CR exhibited a neuronal distribution: about 75% of the spiral ganglion neurons, some nerve fibers in the modulius and nerve fibers running in the basilar membrane were CR-labeled. During the postnatal maturation, a CR-immunostaining appeared around the IHC body and CR was expressed transiently in the OHC. In the adult-like dog cochlea, a CR-positive network surrounded the unlabeled IHC. The neuronal CR-labeling remained unchanged from birth.

Bilateral deafness in a maltese terrier and a great pyrenean puppy: inner ear morphology.

Two puppies, a 4-month-old female Maltese terrier and a 6-week-old male Great Pyrenean, were presented for confirmation of bilateral deafness by electrophysiological testing. In both puppies, brainstem auditory potentials were not evoked by 90 dB NHL click stimulation of each ear. Examination of the inner ear revealed a bilateral cochleo-saccular degeneration in both animals. The lesions were characterized by generalized atrophy of the stria vascularis, collapse of the cochlear duct, degeneration of the organ of Corti, an abnormal tectorial membrane, and saccular collapse, with a normal spiral ganglion. The cochlear duct was entirely obliterated throughout the cochleae in the Maltese terrier puppy, but was locally and asymmetrically affected in the Great Pyrenean. The abnormalities observed in the Maltese terrier puppy were identical with those previously described in deaf Dalmatian puppies; the lesions observed in the Great Pyrenean, however, were less typical. This is the first histopathological description of cochleo-saccular degeneration in the Maltese terrier and Great Pyrenean breeds. In both puppies the defect was probably congenital.