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PubMed1991

Effects of relative starting phase and frequency separation of two-tone stimuli on the brain-stem auditory-evoked response.

Brain-stem auditory-evoked responses (BAERs) were obtained in six normal-hearing adults using single-tone and two-tone stimuli arithmetically centered around 4000 Hz. Two-tone stimuli varied in frequency separation from 200 to 3200 Hz, and started in-phase (homophasic) or 180 deg out-of-phase (antiphasic) with each other. Responses to each of the single-tone components of the two-tone stimuli were elicited and then summed for comparison with responses to the two-tone stimuli. Results indicated no significant difference in wave V latency between homophasic or antiphasic two-tone conditions, and summed single-tone conditions. Under the homophasic condition, the mean latency for the widest frequency separation of the tones was significantly longer than those for narrower separations. A significant difference in wave V amplitude between two-tone phase conditions was found for frequency separations of 200, 400, and 3200 Hz only. Summed single-tone BAERs demonstrated a significantly larger wave V amplitude than responses from either two-tone phase condition at all frequency separations.

Adult↗
PubMed1998

Release from masking due to spatial separation of sources in the identification of nonspeech auditory patterns.

A nonspeech pattern identification task was used to study the role of spatial separation of sources on auditory masking in multisource listening environments. The six frequency patterns forming the signal set were comprised of sequences of eight 60-ms tone bursts. Bursts of masking sounds were played synchronously with the signals. The main variables in the study were (1) the difference in spatial separation in the horizontal plane between signals and maskers and (2) the nature of the masking produced by the maskers. Spatial separation of signal and masker ranged from 0-180 degrees. The maskers were of two types: (1) a sequence of eight 60-ms bursts of Gaussian noise intended to produce predominantly peripherally based "energetic masking" and (2) a sequence of eight 60-ms bursts of eight-tone complexes intended to produce primarily centrally based "informational masking." The results indicated that identification performance improved with increasing separation of signal and masker. The amount of improvement depended upon the type of masker and the center frequency of the signal patterns. Much larger improvements were found for spatial separation of the signal and informational masker than for the signal and energetic masker. This was particularly apparent when the acoustical advantage of the signal-to-noise ratio in the more favorable of the two ears (the ear nearest the signal) was taken into account. The results were interpreted as evidence for an important role of binaural hearing in reducing sound source or message uncertainty and may contribute toward solving the "cocktail party problem."

Adult↗
PubMed1999

The role of perceived spatial separation in the unmasking of speech.

Spatial separation of speech and noise in an anechoic space creates a release from masking that often improves speech intelligibility. However, the masking release is severely reduced in reverberant spaces. This study investigated whether the distinct and separate localization of speech and interference provides any perceptual advantage that, due to the precedence effect, is not degraded by reflections. Listeners' identification of nonsense sentences spoken by a female talker was measured in the presence of either speech-spectrum noise or other sentences spoken by a second female talker. Target and interference stimuli were presented in an anechoic chamber from loudspeakers directly in front and 60 degrees to the right in single-source and precedence-effect (lead-lag) conditions. For speech-spectrum noise, the spatial separation advantage for speech recognition (8 dB) was predictable from articulation index computations based on measured release from masking for narrow-band stimuli. The spatial separation advantage was only 1 dB in the lead-lag condition, despite the fact that a large perceptual separation was produced by the precedence effect. For the female talker interference, a much larger advantage occurred, apparently because informational masking was reduced by differences in perceived locations of target and interference.

Female↗
PubMed2006

Low-field magnetic separation of monodisperse Fe3O4 nanocrystals.

Magnetic separations at very low magnetic field gradients (<100 tesla per meter) can now be applied to diverse problems, such as point-of-use water purification and the simultaneous separation of complex mixtures. High-surface area and monodisperse magnetite (Fe3O4) nanocrystals (NCs) were shown to respond to low fields in a size-dependent fashion. The particles apparently do not act independently in the separation but rather reversibly aggregate through the resulting high-field gradients present at their surfaces. Using the high specific surface area of Fe3O4 NCs that were 12 nanometers in diameter, we reduced the mass of waste associated with arsenic removal from water by orders of magnitude. Additionally, the size dependence of magnetic separation permitted mixtures of 4- and 12-nanometer-sized Fe3O4 NCs to be separated by the application of different magnetic fields.

Journal Article↗
PubMed2006

Dose separation does not overcome the pharmacokinetic interaction between fosamprenavir and lopinavir/ritonavir.

Previous investigations have shown a significant negative two-way drug interaction between fosamprenavir (FPV) and lopinavir/ritonavir (LPV/RTV) in both human immunodeficiency virus (HIV)-infected patients and seronegative volunteers. This randomized, nonblinded, three-way crossover study of HIV-seronegative adult volunteers investigated dose separation and increased doses of RTV as a means to overcome the interaction between FPV and LPV/RTV. Eleven HIV-seronegative volunteers were given FPV plus LPV/RTV at 700 mg plus 400/100 mg every 12 hours (q12h) simultaneously for 10 days and then randomized to receive each of three 7-day treatments in one of six possible sequences, as follows: FPV plus LPV/RTV at 700 mg plus 400 mg/100 mg q12h simultaneously, FPV/RTV at 700 mg/100 mg q12h plus LPV/RTV at 400 mg/100 mg q12h, with doses separated by 4 h, and FPV/RTV at 1,400 mg/200 mg in the morning plus LPV/RTV at 800 mg/200 mg in the evening. Pharmacokinetic sampling was performed on day 8 of each treatment, and samples were analyzed for FPV, amprenavir (APV), LPV, and RTV concentrations by high-performance liquid chromatography-tandem mass spectrometry. Geometric mean ratios (GMR [with 95% confidence intervals]) for the 4- and 12-h dose separation strategies compared to simultaneous administration were calculated for the areas under the concentration-time curves from 0 to 24 h. Compared to simultaneous administration, RTV exposures increased with both 4-h and 12-h dose separation strategies (GMR, 5.30 [3.66 to 7.67] and 4.45 [3.09 to 6.41], respectively). LPV exposures also significantly increased with both 4-h and 12-h dose separation strategies (GMR, 1.76 [1.34 to 2.32] and 1.43 [1.02 to 2.01], respectively). However, both the 4- and 12-h strategies resulted in greater reductions in APV exposure (0.67 [0.54 to 0.83] and 0.77 [0.59 to 0.99], respectively) compared to simultaneous administration. Additional investigations are warranted to determine the optimal dosing of FPV with LPV/RTV.

Adolescent↗
PubMed2006

Fluorescence in situ hybridization-flow cytometry-cell sorting-based method for separation and enrichment of type I and type II methanotroph populations.

A fluorescence in situ hybridization-flow cytometry (FISH/FC)-based method was optimized using artificial mixtures of pure cultures of methanotrophic bacteria. Traditional oligonucleotide probes targeting 16S rRNAs of type I (MG84/705 probe) and type II (MA450 probe) methanotrophs were labeled with fluorescein or Alexa fluor and used for FISH, followed by fluorescence-activated FC analysis and cell sorting (FACS). The method resulted in efficient separation of target cells (type I or type II methanotrophs) from the artificial mixtures. The method was then applied for detection and enrichment of type I and type II methanotroph populations from a natural sample, Lake Washington sediment. Cells were extracted from the sediment, fixed, and subjected to FISH/FC/FACS. The resulting subpopulations were analyzed by reverse transcriptase PCR surveys of 16S rRNA, pmoA (encoding a subunit of particulate methane monooxygenase), and fae (encoding formaldehyde-activating enzyme) genes. The functional gene analysis indicated specific separation of the type I and type II methanotroph populations. 16S rRNA gene analysis revealed that type I methanotrophs comprised 59% of the subpopulation separated using the type I-specific probe and that type II methanotrophs comprised 47.5% of the subpopulation separated using the type II-specific probe. Our data indicate that the FISH/FC/FACS protocol described can provide significant enrichment of microbial populations of interest from complex natural communities and that these can be used for genetic tests. We further tested the possibility of direct whole-genome amplification (WGA) from limited numbers of sorted cells, using artificial mixtures of microbes whose genome sequences are known. We demonstrated that efficient WGA can be achieved using 10(4) or more cells separated by 16S rRNA-specific FISH/FC/FACS, while fewer cells resulted in less specific WGA.

DNA Primers↗
PubMed1971

Separation of mycobacterial soluble antigens by isoelectric focusing.

Ion-exchange chromatography and gel filtration have been reported to yield partial separation of mycobacterial antigens. These procedures were used in combination with isoelectric focusing in an attempt to purify antigens of Mycobacterium tuberculosis strain H37Ra. The fractionating action of isoelectric focusing is dependent upon differences in the isoelectric points of the proteins to be separated. Culture filtrate of M. tuberculosis H37Ra was chromatographed on Sephadex G-200. This resulted in two widely separated peaks. The first peak, presumably containing high-molecular-weight substances, was then fractionated on a diethylaminoethyl Sephadex anion-exchange column. Three peaks were collected, and each was subjected to isoelectric focusing. Each peak was further separated into two or more fractions. The serological reactivity of each fraction was determined by immunodiffusion and immunoelectrophoresis. Sensitized guinea pigs were also skin-tested with the fractions. Two of the fractions contained only a single precipitinogen. One fraction contained two precipitinogens. A fourth fraction contained three precipitinogens and was also the only fraction to display sensitin activity. Four of the fractions were inactive either as precipitinogens or sensitins. The results suggest that the methods described are useful for the separation of mycobacterial antigens.

Journal Article↗
PubMed2006

A new D,L-endopeptidase gene product, YojL (renamed CwlS), plays a role in cell separation with LytE and LytF in Bacillus subtilis.

A new peptidoglycan hydrolase, Bacillus subtilis YojL (cell wall-lytic enzyme associated with cell separation, renamed CwlS), exhibits high amino acid sequence similarity to LytE (CwlF) and LytF (CwlE), which are associated with cell separation. The N-terminal region of CwlS has four tandem repeat regions (LysM repeats) predicted to be a peptidoglycan-binding module. The C-terminal region exhibits high similarity to the cell wall hydrolase domains of LytE and LytF at their C-terminal ends. The C-terminal region of CwlS produced in Escherichia coli could hydrolyze the linkage of d-gamma-glutamyl-meso-diaminopimelic acid in the cell wall of B. subtilis, suggesting that CwlS is a d,l-endopeptidase. beta-Galactosidase fusion experiments and Northern hybridization analysis suggested that the cwlS gene is transcribed during the late vegetative and early stationary phases. A cwlS mutant exhibited a cell shape similar to that of the wild type; however, a lytE lytF cwlS triple mutant exhibited aggregated microfiber formation. Moreover, immunofluorescence microscopy showed that FLAG-tagged CwlS was localized at cell separation sites and cell poles during the late vegetative phase. The localization sites are similar to those of LytF and LytE, indicating that CwlS is involved in cell separation with LytF and LytE. These specific localizations may be dependent on the LysM repeats in their N-terminal domains. The roles of CwlS, LytF, and LytE in cell separation are discussed.

Bacillus subtilis↗
PubMed1974

Cell separation in blue-green bacteria.

Autolysin-like enzymes appear to be responsible for cell separation in Agmenellum quadruplicatum. Mutants that are impaired in cell separation and grow as chains exhibit reduced cell lytic activity. Lysozyme, extracted autolysin, and antibiotics that affect peptidoglycan synthesis phenotypically suppress chain formation. Various aspects of the regulation of the cell separation process were also examined. Studies involving antibiotic inhibitors of macromolecular synthesis and general growth inhibitors provided no evidence for the active regulation of the cell separation process during the latter portion of the division cycle. Evidence was obtained, however, for the partial restriction of peptidogly-can hydrolysis by unknown secondary modifications. The thin electron-dense layer of peptidoglycan along the sides of cells was much more resistant to hydrolysis by egg-white lysozyme than was the septum between daughter cells. The middle portion of the septum was more sensitive than was the layer immediately adjacent to the cytoplasmic membrane. Under conditions that would not osmotically stabilize spheroplasts, lysozyme facilitates rapid cell separation in chain-forming mutants with little leakage of cellular protein or loss of viability.

Anti-Bacterial Agents↗
PubMed2001

The Cbk1p pathway is important for polarized cell growth and cell separation in Saccharomyces cerevisiae.

During the early stages of budding, cell wall remodeling and polarized secretion are concentrated at the bud tip (apical growth). The CBK1 gene, encoding a putative serine/threonine protein kinase, was identified in a screen designed to isolate mutations that affect apical growth. Analysis of cbk1Delta cells reveals that Cbk1p is required for efficient apical growth, proper mating projection morphology, bipolar bud site selection in diploid cells, and cell separation. Epitope-tagged Cbk1p localizes to both sides of the bud neck in late anaphase, just prior to cell separation. CBK1 and another gene, HYM1, were previously identified in a screen for genes involved in transcriptional repression and proposed to function in the same pathway. Deletion of HYM1 causes phenotypes similar to those observed in cbk1Delta cells and disrupts the bud neck localization of Cbk1p. Whole-genome transcriptional analysis of cbk1Delta suggests that the kinase regulates the expression of a number of genes with cell wall-related functions, including two genes required for efficient cell separation: the chitinase-encoding gene CTS1 and the glucanase-encoding gene SCW11. The Ace2p transcription factor is required for expression of CTS1 and has been shown to physically interact with Cbk1p. Analysis of ace2Delta cells reveals that Ace2p is required for cell separation but not for polarized growth. Our results suggest that Cbk1p and Hym1p function to regulate two distinct cell morphogenesis pathways: an ACE2-independent pathway that is required for efficient apical growth and mating projection formation and an ACE2-dependent pathway that is required for efficient cell separation following cytokinesis. Cbk1p is most closely related to the Neurospora crassa Cot-1; Schizosaccharomyces pombe Orb6; Caenorhabditis elegans, Drosophila, and human Ndr; and Drosophila and mammalian WARTS/LATS kinases. Many Cbk1-related kinases have been shown to regulate cellular morphology.

Amino Acid Sequence↗
PubMed2004

A comparison of IgG anti-rubella activity in frozen serum stored in primary gel separation tubes or secondary tubes.

AIMS: To determine the suitability of primary gel separation tubes for the storage of frozen sera intended for serological testing. METHODS: Blood samples from 102 patients were collected into gel separation tubes. The sera from these samples were split between the primary gel separation tubes and conventional plastic storage tubes and frozen. A year later, the tubes were thawed and anti-rubella IgG concentrations were compared for the serum pairs using the Wilcoxon signed rank test. RESULTS: No significant difference was detected between the two storage methods. CONCLUSIONS: Frozen storage of serum samples in primary gel separation tubes is a practical alternative to storing separated sera in secondary containers. Adopting this practice has advantages for laboratories in reducing specimen handling and reducing errors in labelling stored samples.

Adult↗
PubMed2000

A semi-parametric hybrid neural model for nonlinear blind signal separation.

Nonlinear blind signal separation is an important but rather difficult problem. Any general nonlinear independent component analysis algorithm for such a problem should specify which solution it tries to find. Several recent neural networks for separating the post nonlinear blind mixtures are limited to the diagonal nonlinearity, where there is no cross-channel nonlinearity. In this paper, a new semi-parametric hybrid neural network is proposed to separate the post nonlinearly mixed blind signals where cross-channel disturbance is included. This hybrid network consists of two cascading modules, which are a neural nonlinear module for approximating the post nonlinearity and a linear module for separating the predicted linear blind mixtures. The nonlinear module is a semi-parametric expansion made up of two sub-networks, one of which is a linear model and the other of which is a three-layer perceptron. These two sub-networks together produce a "weak" nonlinear operator and can approach relatively strong nonlinearity by tuning parameters. A batch learning algorithm based on the entropy maximization and the gradient descent method is deduced. This model is successfully applied to a blind signal separation problem with two sources. Our simulation results indicate that this hybrid model can effectively approach the cross-channel post nonlinearity and achieve a good visual quality as well as a high signal-to-noise ratio in some cases.

Algorithms↗
PubMed1996

Are MR imaging signs of meniscocapsular separation valid?

PURPOSE: To define the positive predictive value (PPV) for the magnetic resonance (MR) imaging diagnosis of meniscocapsular separation in the knee. MATERIALS AND METHODS: The MR reports of 52 patients aged 16-75 years who had a prospective MR diagnosis of meniscocapsular injury were correlated with arthroscopic results. MR images obtained in 50 of these patients were retrospectively reviewed for specific signs of meniscocapsular separation, which included meniscal displacement, peripheral meniscal corner tears, increased perimeniscal signal intensity, fluid deep to the medial collateral ligament, and abnormal lateral fascicles. MR findings were correlated with arthroscopic findings. RESULTS: For the prospective MR interpretations, the PPV for meniscocapsular separation was 9% medially and 13% laterally. Meniscal displacement (measured from the meniscal edge to the tibia) was as great as 10 mm medially or 13 mm laterally without meniscocapsular tears at arthroscopy. Meniscal displacement did not correlate with effusion. Meniscal corner tears had a PPV of 0% medially and 50% laterally. Fluid at the meniscocapsular border and fluid deep to the medial collateral ligament were poor predictors of meniscocapsular abnormalities. Abnormal-appearing meniscal fascicles had a PPV of 8% for lateral meniscocapsular separation. CONCLUSION: The PPV for the MR diagnosis of meniscocapsular separation is low; the reported MR signs correlate poorly with arthroscopic findings.

Adolescent↗
PubMed1988

Separation of cardiac myosin heavy chains by gradient SDS-PAGE.

Separation of alpha- and beta-myosin heavy chains (MHCs) in cardiac ventricles of rats by gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was accomplished and compared with the separation of myosin isozymes obtained with pyrophosphate gels. Whole muscle homogenates were electrophoresed on a 4-9% linear gradient SDS polyacrylamide gel for 3-4 h. MHC bands were identified by the migration distance relative to a MHC standard and immunoblot results with a monoclonal antibody to MHC. The MHC bands were further identified as alpha and beta based on the electrophoretic mobility of ventricular homogenates from hypothyroid and hyperthyroid rats and ventricular and slow soleus skeletal muscle homogenates from control rats. The beta-MHC migrated faster than alpha-MHC, and laser densitometry revealed separate peaks when both MHCs were present. With homogenates containing MHC ranging from 0 to 100% alpha, the separation of MHCs with gradient SDS-PAGE correlated highly (r = 0.97) with separation of myosin isozymes by pyrophosphate gel electrophoresis. The SDS-PAGE technique reported herein is a quick, valid, and direct method for the identification and quantification of ventricular MHCs.

Animals↗
PubMed1998

Electrophoretic separation and quantitation of cardiac myosin heavy chain isoforms in eight mammalian species.

A protocol for sample preparation and gel electrophoresis is described that reliably results in the separation of the alpha- and beta-isoforms of cardiac myosin heavy chain (MHC-alpha and MHC-beta) in eight mammalian species. The protocol is based on a simple, nongradient denaturing gel. The magnitude of separation of MHC-alpha and MHC-beta achieved with this protocol is sufficient for quantitative determination of the relative amounts of these two isoforms in mouse, rat, guinea pig, rabbit, canine, pig, baboon, and human myocardial samples. The sensitivity of the protocol is sufficient for the detection of MHC isoforms in samples at least as small as 1 microgram. The glycerol concentration in the separating gel is an important factor for successfully separating MHC-alpha and MHC-beta in myocardial samples from different species. The effect of sample load on MHC-alpha and MHC-beta band resolution is illustrated. The results also indicate that inclusion of a homogenization step during sample preparation increases the amount of MHC detected on the gel for cardiac samples to a much greater extent than for skeletal muscle samples. Although the protocol described in this study is excellent for analyzing cardiac samples, it should be noted that the same protocol is not optimal for separating MHC isoforms expressed in skeletal muscle, as is illustrated.

Animals↗
PubMed1995

Chemical sympathectomy and maternal separation affect neonatal stress responses and adrenal sensitivity to ACTH.

The participation of sympathetic adrenal innervation in the control of the neonatal adrenocortical system and in changes in adrenal sensitivity after maternal separation for 24 h was tested in 10- and 23-day-old pups. Chemical sympathectomy by guanethidine (20 mg/kg body wt) decreased basal and stimulated corticosterone compound B (B) secretion without affecting adrenocorticotropic hormone (ACTH) release, abolished the enhanced adrenal sensitivity to ACTH induced by maternal separation in 10-day-old pups, but did not modify adrenal sensitivity following ether stress in 23-day-old pups. Guanethidine treatment did not affect body and adrenal weight or adrenal choline acetyltransferase activity, but it increased tyrosine hydroxylase activity at both ages. Both chronic guanethidine treatment and acute corticotropin-releasing factor immunoneutralization reduced plasma B levels after maternal separation without affecting plasma ACTH levels. Maternal separation in 10-day-old pups enhanced basal and stimulated ACTH and B secretion after exposure to ether vapors and insulin-induced hypoglycemia (IIH). In nonseparated pups, IIH did not stimulate ACTH secretion and caused small increases in B secretion; however, the enhanced response of separated pups to IIH was due to the effects of intraperitoneal injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Glands↗
PubMed1993

Electrophoretic separation of rat skeletal muscle myosin heavy-chain isoforms.

A new technique for the sodium dodecyl sulfate-polyacrylamide gel electrophoretic separation of rat skeletal muscle myosin heavy-chain (MHC) isoforms is presented. This technique allows for the separation of the four identified MHC isoforms known to be present in adult rat skeletal muscle. These types of MHC are commonly called I, IIa, IIx or IId, and IIb. The procedure can be performed using minigel electrophoresis systems and does not involve preparation of gradient-separating gels or the use of special cooling devices. The procedure accommodates both silver and Coomasie Blue staining. Thus the procedure is simple to perform and highly repeatable, providing high-resolution separation of MHC protein isoforms. The percent composition of the four adult MHCs in rat soleus, medial gastrocnemius, diaphragm, and levator ani muscles by use of this procedure and Coomasie Blue staining is similar to that previously reported. This new technique provides a novel and easy-to-perform method for the separation of rat skeletal muscle MHC isoforms.

Animals↗
PubMed2005

Comparisons of serum somatotropin, 3,5,3'-triiodothyronine, thyroxine, total protein and free fatty acid levels in newborn Sakiz lambs separated from or suckling their dams.

OBJECTIVE: To determine the effects on serum somatotropin, 3,5,3'-triiodothyronine (T(3)), thyroxine (T(4)), free fatty acids (FFAs) and total protein levels of different feedings and age in Sakiz sheep that have a high twin-bearing rate supported by estrous synchronization. METHODS: 20 newborn lambs were used in the study. Lambs were divided into 2 equal groups. The lambs in 1 group were separated from their dams following parturition, and those in other group were kept together with their dams. Separated lambs were fed commercial cow's milk for 2.5 weeks. After that, they were fed a milk substitute, hay and concentrated supplement for 2 months, and from the end of the 2 months they were fed hay and concentrated supplement. The lambs in other group were kept together with their dams only during the day and in addition were fed hay and concentrated supplement. After 2 months, they were maintained feeding only on hay and concentrated supplement. Blood samples were taken from vena jugularis of lambs at 12 and 24-48 h, and 14, 28, 42 and 56-90 days after parturition. Serum samples were analyzed for somatotropin by enzyme immunoassay, for T(3) and T(4) by radioimmunoassay, and for total protein and FFA by a spectrophotometric method. RESULTS: Serum somatotropin levels were insignificantly different between the lambs with and without their dams. Serum T(3) levels were generally lower in the separated lambs than those in other group. Serum T(4) levels were significantly lower in the separated lambs 48 h and 90 days after parturition than those in other group. Serum FFAs were insignificantly different between 2 groups. Serum total protein concentrations were lower in the separated lambs than lambs kept together with their dams (significantly at 12 and 48 h and 14 days). CONCLUSION: Ingestion of colostrum and dam milk markedly affected serum total protein levels, but body weight and serum somatotropin levels were insignificantly affected. Also serum T(3) and T(4) levels markedly decreased with advancing age. Serum FFA levels were not affected by the trial.

Animal Feed↗