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Monocular deprivation (MD) during the critical period reduces the visual cortical response to the deprived eye and causes the geniculocortical axons serving the deprived eye to retract. When MD is combined with a pharmacological inhibition of the visual cortex, the cortical neurons weaken their response to an open eye and the input axons serving the open eye retract. To determine whether the 2 types of ocular dominance (OD) plasticity reflect an experience-driven modification of neural circuits sharing the same developmental time course, we analyzed the OD plasticity in an inhibited visual cortex using cats at different ages. MD did not affect the OD distribution in the inhibited cortex of adults, confirming that the OD plasticity in the inhibited cortex represents a developmental plasticity. In developing animals, the OD plasticity in the inhibited cortex was observed at the late phase of the critical period (P40–46) but not at the early phase (P22–26). We found a retraction of input axons serving an open eye at the late phase, whereas those at the early phase were comparable to the axons of normal animals. Therefore, the maturation of visual circuits might include an experience-driven rearrangement of thalamocortical projections during the late phase of development.
In the mammalian visual system, monocular deprivation (MD) during development causes the loss of cortical responses to the deprived eye (ocular dominance [OD] plasticity) (Wiesel and Hubel 1963). Anatomically, afferent axons from the lateral geniculate nucleus (LGN) serving the deprived eye retract (Antonini and Stryker 1993, 1996) and ultimately lose their cortical territory (Shatz and Stryker 1978; LeVay et al. 1980).
Previous works have demonstrated the role of the activity of cortical cells in OD plasticity. When MD is combined with a pharmacological inhibition of the visual cortical neurons by administering muscimol, a GABAA receptor agonist, leaving the activity of the geniculocortical afferents intact, the cortical neurons weaken their responses to the open eye and active afferents serving the open eye selectively retract (Reiter and Stryker 1988; Hata and Stryker 1994; Hata et al. 1999).
Although the 2 types of OD plasticity, one in the normal cortex and the other in the inhibited cortex, both reflect an experience-driven modification of neural circuits, previous studies demonstrated their distinct characteristics. An imbalance of inputs from the 2 eyes is necessary for OD plasticity in the normal cortex because the cortical responses and input axons remain mostly intact after binocular deprivation (Wiesel and Hubel 1965; Antonini and Stryker 1998). In contrast, in the inhibited cortex, the afferents serving an open eye significantly retract, regardless of whether the other eye is closed (Haruta and Hata 2007). In addition, OD plasticity in the normal cortex requires the activation of an intracellular signaling cascade involving the cAMP/PKA pathway (Beaver et al. 2001; Fischer et al. 2004), whereas OD plasticity in the inhibited cortex can be induced in the presence of a PKA inhibitor (Shimegi et al. 2003). These studies suggest that the 2 types of OD plasticity may represent distinct components of the experience-dependent development of the visual system. Alternatively, however, the reverse OD plasticity in the artificially inhibited cortex might be caused by an unknown mechanism that is not related to developmental plasticity. Therefore, it is important to determine whether the reverse OD plasticity in the inhibited cortex operates only in developing animals.
In the normal cortex, MD affects visual responsiveness in a restricted period of development called the critical period with its peak at the fourth week of age in cats (Olson and Freeman 1980). Therefore, we analyzed the OD of cortical neurons and morphology of geniculocortical axons in a pharmacologically inhibited visual cortex using cats at around the peak of the critical period (early phase, postnatal days [P] 22–26), at the late phase of this period (P40–46) and in adults to determine whether the OD plasticity in the normal and inhibited cortices shares the same developmental time course.
We demonstrate that the reverse OD plasticity is not observed in adult animals, confirming that the plasticity in the inhibited cortex reflects an aspect of developmental plasticity. Furthermore, we demonstrate that the reverse OD plasticity operates only in the late phase of the critical period but not at the peak of it. These observations suggest that the critical period may consist of multiple stages that have distinct time course.
Materials and Methods
All of the cats in this study were born in the breeding colony of Tottori University. The experimental procedures used met the guidelines of the animal care committee of Tottori University.
A stainless steel cannula connected to an osmotic minipump (Alzet 2001 or 2002, Durect, CA, United States of America) was implanted in the left hemisphere of the primary visual cortex (stereotaxic location: A −3.0 mm, L 2.0 mm, depth from cortical surface 2.0 mm) of animals of either sex at the early phase (P22–26) and the late phase (P40–46) of the critical period and adults (>P500), and a muscimol solution was infused continuously (30 mM in Ringer's solution, 1.0 or 0.5 µL/h, Sigma-Aldrich, MO, United States of America or Tocris Bioscience, MO, United States of America). During the same surgery, biotinylated dextran amine (BDA-10 000, Molecular Probes, OR, United States of America) was injected at lamina A of the LGN to label the geniculocortical axons. A glass pipette filled with BDA solution was inserted stereotaxically to locate a clear visual response from lamina A of the LGN, which is ipsilateral to the infusion cannula. BDA was iontophoretically injected (pipette positive current of 3–5 µA, 2 s on/off pulse, 50–70 times) at 3–5 sites in each LGN. After 1–2 days, either eye was deprived of vision by eyelid suture so that the labeled LGN axons represent inputs from either an open or closed eye. BDA was also injected in several normal animals at P22–23 and P41–43. All of the surgical procedures were performed using sterile conditions under anesthesia with 1.5–3.5% isoflurane in a 1:1 mixture of N2O and O2. All of the incisions were infiltrated with local anesthetics (Xylocaine pump spray, AstraZeneca, Japan). The animals were given an antimicrobial agent (enrofloxacin, 5 mg/kg, Baytril, Bayer, Germany) every day after the surgery.
Following 6 days of MD (7 or 8 days of muscimol infusion) or at the corresponding age in the case of the normal animals, the single-unit activity was recorded from the primary visual cortex (early phase, P28–34; late phase, P47–53). Anesthesia was induced with 4–5% isoflurane in N2O and O2 (1:1) and maintained with pentobarbital sodium (2–4 mg/kg/h i.v., Nembutal, Dainippon Sumitomo Pharma, Japan). The pupils were dilated with 0.5% tropicamide and 0.5% phenylephrine hydrochloride (Mydrin-P, Santen Pharma, Japan), and contact lenses were placed on the corneas. The animals were paralyzed with gallamine triethiodide (10 mg/kg/h, i.v., Sigma-Aldrich) or pancuronium bromide (0.2 mg/kg/h, i.v., Mioblock, Nihon Organon, Japan) and maintained under artificial respiration. The end-tidal CO2 concentration and body temperature were maintained at 4% and 38°C, respectively. All of the incisions were infiltrated with local anesthetics. The cortical area that was inhibited by muscimol was delineated by recording the cortical cell activity at various distances to the infusion cannula using a tungsten microelectrode, and then the pump was removed. After the recovery of cortical activity from the inhibitory effect of muscimol, the visual response of cortical neurons was recorded, and the OD of individual neurons was determined in both hemispheres on the basis of a conventional 7-point classification (Hubel and Wiesel 1962).
The OD distribution of each hemisphere was quantified by calculating the contralateral-eye bias index (CBI) as follows:
After the recording, a needle mark was made in the cortex at the border of the inhibited region. In the normal animals, 2 needle marks were made with 1 mm separation. The needle marks and cannula tracks were used to calculate the shrinkage correction in the analysis of the axon morphology. The animals were euthanized with an overdose of pentobarbital sodium (100 mg/kg, i.v.) and perfused transcardially with cold phosphate-buffered saline and 4% paraformaldehyde in 0.1 M phosphate buffer. Tissue blocks containing the LGN and entire caudal pole of the cortex were cut using a freezing microtome in the frontal plane (50 µm thickness). All of the sections were processed using the standard ABC kit (Vector Laboratories, CA, United States of America) to visualize the tracer. Selected sections containing the visual cortex or LGN were stained with cresyl violet for the localization of the layer boundaries and injection sites.
Axonal Arbor Analysis
All of the injection sites were well confined to lamina A of the LGN (Fig. 2A,B). When an injection site invades the lamina A1 representing the ipsilateral eye, we eliminated all of the samples from the same animal from the arbor reconstruction. The BDA-filled arbors were reconstructed from serial sections in 3 dimensions with the aid of computer software (Neurolucida, Microbrightfield, VT, United States of America) and then corrected for tissue shrinkage. We calculated 2 measures, the total length and branch point number, to quantify the size and complexity of the individual axonal arbors. The total length was obtained by adding the length of all of the axonal segments constituting the terminal field of an arbor. The branch point number is the number of axon bifurcations in the terminal arborization. Only the portion of the arbor located in layers II/III and IV was considered for these analyses. Very short endings <5 µm were omitted from the analyses.
Age-Dependence of the Physiological OD Plasticity in the Inhibited Visual Cortex
We infused one hemisphere of the visual cortex of young (early phase of the critical period, P22–26; late phase of the critical period, P40–46) and adult (>P500) cats with muscimol, and they were monocularly deprived for 6 days. MD caused a loss of visual response to the deprived eye in the control hemisphere, which is the untreated hemisphere contralateral to the muscimol-infusion site, of kittens at both the early and the late phase of the critical period but not in adults (Fig. 1A, "MD"), as demonstrated previously (Olson and Freeman 1980). We calculated the CBI (see the Materials and Methods section) for the individual hemispheres to evaluate the degree of OD plasticity. For the normal animals, the CBI values were 0.53 ± 0.05 (mean ± SD, n = 3 animals) in the early-phase group and 0.54 ± 0.04 (n = 4) in the late-phase group, reflecting a slight natural bias toward the contralateral eye (Fig. 1A, "normal"). The CBIs of the control hemispheres of young animals (early, 0.09 ± 0.06; late, 0.08 ± 0.04) were significantly deviated toward the ipsilateral open eye versus those of the normal animals in the 2 age groups (Fig. 1A, "MD", and Fig. 1B).
In the inhibited cortex, the cortical neurons responded preferentially to the ipsilateral deprived eye (reverse OD plasticity) in the kittens at the late phase of the critical period (Fig. 1A, "Late", "MD + muscimol"). Their CBIs (mean ± SD = 0.43 ± 0.10) were significantly different from those of the control hemisphere, showing that the OD shift toward an open eye had been blocked (Fig. 1B, "Late"). Furthermore, the CBIs in the inhibited cortex were significantly deviated from those of the normal animals toward the deprived eye (Fig. 1B, "Late"). In the kittens at the early phase of the critical period, most neurons in the inhibited cortex responded to both eyes equally (Fig. 1A, "Early", "MD + muscimol"). Their CBIs (0.51 ± 0.03) were significantly higher than those of the control hemisphere, showing a blockage of the OD shift toward an open eye (Fig. 1B, "Early"). Unlike the late-phase group, however, the OD distribution in the inhibited cortex did not show a shift toward the deprived eye and was comparable to that of the normal animals (Fig. 1B, "Early"). The direct comparison of the OD distribution in the inhibited cortex showed a significant difference between the early- and late-phase groups (Fig. 1A, early vs. late in "MD + muscimol", P < 0.05, χ2 test).
The adult animals preserved the binocular responses in both the control (Fig. 1, "MD", CBI = 0.55 ± 0.04) and the inhibited cortices ("MD + muscimol", CBI = 0.53 ± 0.04), confirming that the reverse OD plasticity can be expressed only in the developing brain, similar to the OD plasticity in the intact cortex. Therefore, the reverse OD plasticity in the inhibited cortex should reflect a type of developmental plasticity that operates only in the late phase of the critical period.
Experience-Driven Retraction of Geniculocortical Axons in the Late Phase of the Critical Period
Previous studies demonstrated that input axons from the LGN carrying information from an open eye significantly retract in the inhibited cortex (Hata et al. 1999; Haruta and Hata 2007). To determine the age dependence of the structural remodeling of input axons, we injected an anterograde tracer, BDA, into lamina A of the LGN and deprived vision in the ipsilateral eye, so as to label the axons serving the open eye (Fig. 2A,B). The cortical arbors of the geniculocortical axons in the inhibited cortex were labeled well with BDA, as shown in Figure 2C,D. We found no noticeable difference in the quality of labeling between the axons in the 2 age groups.
Axonal arbors serving the open eye in the inhibited cortical region were reconstructed from the animals at the early phase (n = 18 arbors from 8 animals) and the late phase (n = 11 arbors from 4 animals) of the critical period and compared with those of the age-matched normal animals (early, n = 9 arbors from 3 animals; late, n = 6 arbors from 2 animals). Examples of arbors from each group are shown in Figure 2E,F. We quantitatively evaluated the morphology of the individual axons by measuring 2 parameters: The total length of axonal segments and the number of branch points of the cortical arbors (Fig. 3). In the animals at the late phase of the critical period, the open-eye arbors in the inhibited cortex were significantly shorter and had fewer branch points than those in the age-matched normal animals (total length, P < 0.005; branch points, P < 0.01, Student's t-test) (Fig. 3, "Late").However, the open-eye arbors in the inhibited cortex at the early phase of the critical period were preserved and similar in total length and branching number to those in the normal animals (not significant, Student's t-test) (Fig. 3, "Early"). We had previously reported that the arbors serving the deprived eye in the muscimol-treated cortex do not show a retraction and are comparable to normal axons in animals at the late phase of the critical period (Hata and Stryker 1994; Haruta and Hata 2007). Thus, we further determined whether visual inputs influence axonal morphology in the inhibited cortex of animals at the early phase of the critical period. By labeling the LGN neurons in lamina A and depriving the contralateral eye of vision, we reconstructed the deprived-eye arbors in the muscimol-treated cortex of early-age animals (10 arbors from 3 animals; Fig. 2E). We found no significant difference in the number of branching or total length between the deprived-eye arbors and open-eye arbors in the muscimol-treated cortex or between the deprived-eye arbors in the inhibited cortex and normal arbors (not significant, Student's t-test; Fig. 3, "Early"). Therefore, experience-dependent retraction of LGN axons in the inhibited cortex should occur at the late phase of the critical period but not around the peak of it.
In this study, the reverse OD plasticity in the inhibited cortex was observed only at the late phase of the critical period. Consistently, we found a retraction of the LGN axons serving an open eye in the inhibited cortex at the late critical period, whereas the axons at the early phase were comparable to those in the age-matched normal animals. Three previous studies have demonstrated reverse OD plasticity in the inhibited cortex (Reiter and Stryker 1988; Hata and Stryker 1994; Shimegi et al. 2003). The lack of reverse OD plasticity in the early-phase group of the present study does not contradict the previous results because those studies had used older animals (age at MD: P30–35 [Reiter and Stryker 1988], P30–37 [Hata and Stryker 1994], and P43–45 [Shimegi et al. 2003]). Among them, Reiter and Stryker (1988) had begun the MD at P30–35 and recorded at P35–43, whereas we recorded in the early-phase group of the present study at P30–34, which corresponds to the age of the beginning of MD in the previous study. Thus, the lack of reverse OD plasticity in the early-phase group in the present study suggests that the beginning of the period of reverse OD plasticity would be later than P30. Regarding the other 2 previous studies, Hata and Stryker (1994) had used a longer MD (4 weeks from P30 to 37) which completely covered the MD period of the late-phase group, and Shimegi et al. (2003) had used a similar protocol as the present late-phase group (MD from P43–45 to P48–50). Therefore, reverse OD plasticity can be observed until approximately P50, though we do not assert that the period terminates at approximately P50. Although the OD plasticity of cats declines gradually with age and the age of termination is not precisely determined, it is possible that we could observe reverse OD plasticity in animals older than those in the present study, considering that the OD plasticity in the normal cortex can be observed even at 1 year of age (Daw et al. 1992).
OD plasticity is observed in many species including rodents, carnivores, and primates. In the mouse, Gordon and Stryker (1996) defined the critical period of OD plasticity as P19–32 with its peak at around P28 using single-unit analysis, which is comparable to the critical period in rats (Fagiolini et al. 1994). However, subsequent studies using visually evoked potentials to evaluate OD demonstrated that some degree of OD plasticity can be observed in older animals up to P55 in rats and even in adults in mice (Guire et al. 1999; Sawtell et al. 2003). Also, geniculocortical axons extend and elaborate their cortical arbors in experience-dependent manner even after P40 in mice (Antonini et al. 1999). Therefore, some aspects of OD plasticity can proceed in the late- or postcritical period of rodents. On the other hand, the visual system of monkeys appears to develop earlier than that of cats and rodents. Anatomically, the OD columns are formed in utero and appear like adult at birth (Rakic 1976, 1977; Horton and Hocking 1996), suggesting an important role of spontaneous activity in the OD column formation. Postnatally, however, visually driven activity modifies the visual neural circuits. MD just after birth induces OD shift and a change in OD column width (LeVay et al. 1980; Horton and Hocking 1997). Therefore, the critical period appears to begin with no significant delay after birth in the monkey. Assessed by the effect of MD on the OD columns, the critical period of monkeys is considered to last until 10–12 weeks of age (LeVay et al. 1980; Horton and Hocking 1997).
The absence of reverse OD plasticity at the early phase of the critical period may simply reflect a less effective pharmacological inhibition in those animals than in the older ones because the degree of reverse OD plasticity is reported to be proportional to the size of the inhibited cortical area (Reiter and Stryker 1988). In the present study, we found no significant difference between the early- and late-age groups with regard to the size of the inhibited cortical area (early, 4.92 ± 3.19 mm; late, 5.66 ± 1.74 mm, Student's t-test) or in the distance from the cannula to the recording sites (mean ± SD: Early, 1.25 ± 0.36 mm; late, 1.14 ± 0.50 mm, Student's t-test). In addition, we found no significant difference among the arbor groups in the inhibited cortex with regard to the distance from the cannula to the reconstructed arbors (early open, 1.05 ± 0.88 mm; early closed, 1.59 ± 1.09 mm; late open, 1.49 ± 0.86 mm, Student's t-test).
Alternatively, the cortical neurons at the early phase of the critical period may be less sensitive to muscimol than those at the late phase due to an immaturity of the GABAergic inhibitory mechanisms. Actually, the expression levels of GABAA receptors and GABA-synthesizing enzymes increase during early postnatal development in the cat visual cortex (Shaw et al. 1986; Guo et al. 1997). Furthermore, the subunit composition of the GABAA receptor changes to the mature type, which contains the α1 subunit during the critical period of the cat visual cortex (Chen et al. 2001). Therefore, we examined the effect of muscimol on the activity of the visual cortical neurons using an animal at P19, which is younger than the age at the start of the muscimol infusion of the early-phase group. We observed no spontaneous or visually evoked activity in the region around the muscimol infusion site 2 h after starting the infusion. This result is consistent with a previous report showing that an iontophoretic application of GABA inhibited visual cortical activity even in kittens at 6–13 days old (Sato and Tsumoto 1984). Thus, the muscimol application in the present study should have inhibited the cortical activity effectively at both early and late phases of the critical period.
In this study, neither the normal nor reverse OD plasticity was observed in the adults. Thus, both types of OD plasticity should represent a developmental plasticity of the visual cortex. In the developing animals, the OD shift toward an open eye had been blocked in the inhibited cortex of the both early- and late-age animals, suggesting that cortical activity is necessary for the induction of normal OD plasticity in favor of the open eye. However, reverse OD plasticity was observed only at the late phase of the critical period. Considering that PKA activation is necessary for the OD plasticity in the normal cortex (Beaver et al. 2001; Fischer et al. 2004), but not for the reverse OD plasticity in the inhibited cortex (Shimegi et al. 2003), the mechanisms of reverse OD plasticity would be distinct from those of the normal OD plasticity and operate selectively at the late phase of the critical period. Interestingly, several genes encoding synaptic and signaling molecules are regulated by visual experience just after the end of the physiologically defined critical period in the mouse visual cortex (Majdan and Shatz 2006).
Regarding axonal rearrangement, the activity of the cortical neurons should be crucial for the retraction of the axons serving the deprived eye in the normal cortex because the deprived-eye axons retract when their target neurons are activated by visual inputs from the other eye but not when they are inhibited pharmacologically (Hata et al. 1999) or both eyes are deprived of vision (Antonini and Stryker 1998). Conversely, in the inhibited cortex, the afferent axons serving an open eye retract, regardless of whether the other eye is closed (Haruta and Hata 2007). Furthermore, presynaptic mechanisms should mainly underlie the retraction of open-eye axons in the inhibited cortex. Geniculocortical axons exhibit an experience-driven retraction even in the cortex treated with botulinum toxin which prevents synaptic transmission (Watanabe et al. 2009). Thus, the activity of afferent axons may directly regulate their own retraction. Although the mechanism of axon retraction in the inhibited cortex is yet unknown, the visually driven activity might recruit intracellular signals leading to morphological changes such as Ca2+ influx at the terminals, which can regulate axonal behavior (Henley and Poo 2004).
Taken together, the retraction of the open-eye axons in the inhibited cortex represents a developmental mechanism that prunes axons when there is no postsynaptic response. Such a mechanism is suitable for maintaining the OD columns by pruning the axon branches serving the wrong eye, as demonstrated in the eye-specific segregation of retinogeniculate axons (Chapman 2000). It is noteworthy that a large-scale reorganization of the orientation and OD columns occurs in the intact kitten visual cortex at the late phase of the critical period (after postnatal week 7) (Kaschube et al. 2009). Therefore, the structural reorganization of neural circuits, including thalamocortical projections, may occur during the late period of postnatal maturation by pruning the inappropriate connections.
This work was supported by a Grant-in-Aid for Scientific Research on Innovative Areas—Mesoscopic neurocircuitry—from the Ministry of Education, Culture, Sports, Science and Technology of Japan (22115010 to Y.H.).
Conflict of Interest: None declared.


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Hlaudi Motsoeneng says all was well between SABC executives last week.
JOHANNESBURG – The South African Broadcasting Corporation (SABC) COO Hlaudi Motsoeneng says he doesn't know why all controversies at the SABC are blamed on him when the public broadcaster is run by a team.
He says the team that makes decisions at the broadcaster including then acting CEO Jimi Matthews, who resigned this week.
In his resignation letter, Matthews apologised for his part in decisions made at the SABC, saying he can no longer be part of what is happening at the broadcaster.
But Motsoeneng says all was well between SABC executives last week and he's demanding answers about what went wrong to lead to Matthews's resignation.
"We were sitting together on Friday dealing with issues within the organisation, happy. But you ask yourself 'what happened over the weekend? Who said what to who?' Because we're aware that there's an agenda to destabilise this organisation."
Meanwhile, SABC chairman Professor Mbulaheni Maguvhe says the public broadcaster is not in crisis and insists Motsoeneng is doing a sterling job.
An urgent staff meeting took place at the SABC in Johannesburg this afternoon, where James Aguma was appointed as the corporation's acting Chief Executive Officer (CEO).
He's been appointed just a day after Matthews's resignation and three senior reporters spoke out about their working conditions following a series of controversial policy decisions taken by Motsoeneng.
Maguvhe says the SABC is not in crisis and he has commended Motsoeneng for what he describes as "his sterling work", in terms of securing the necessary funds in order for the public broadcaster to carry out its mandate.
The board chairperson says disciplinary processes will continue, if staff members do not adhere to the company's policy.
He says people will come and go, and they are confident that whoever resigns can be replaced by competent individuals.

subset: commoncrawl


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Recently, over at the Audio Daily Blog, the voice-over audio production blog at Voices.com, there was a post with suggestions about how to remove breaths from voice recordings. I figured I'd add my primary method for doing this to their list of 6 tips.
I typically record all my audio into Reaper, an outstanding multi-track recording program that rivals Pro Tools for a fraction of the cost. With music, and with move complex voice-over projects, I'll do all my mixing in Reaper. For voice-over jobs that require multiple parts – say a conversation between two or more people or a series of phrases that need exactly 3 seconds between them (I have done a lot of both types of jobs), Reaper is the tool I use to get everything in the right order and sounding good.
But after I create a single file by mixing it all down in Reaper ("rendering"), I open that single file in a different program – an audio editor – to do the final polish. Some folks consider this overkill, since so much can be done from within Reaper. But coming from a music recording background, I'm used to mastering my mixed-down/rendered audio. And to do that I use a separate audio recording software type called and audio editor. In my case, I use Adobe Audition. But there are much more affordable editors out there.
So once I open the rendered, "almost-finished" audio in my editor, I put the headphones on so I can hear every little breath, click, pop, etc. My primary tool within Adobe Audition for this is called the "Mute" tool (which is also available in the free audio editor called Audacity, BTW, called the "Silence" tool). This tool removes all audio in a selected area, which is different from "deleting it!" If you delete the audio, you also delete that section of time. Silencing leaves the time alone, simply making it audio-free time.go through the audio, selecting breaths, and then silencing them with that silence tool.
Anyway, I actually /mute silence all the sections between phrases to get rid of extraneous little noises that become audible when there is no vocal, including lots of the breaths. It works really well, and its fast.
Heed this warning though. A lot of breaths are just part of the way we speak, especially the breaths before a phrase (both my wife and I tend to put a little exhale or quick intake of breath sound at the end of our phrases. Most of these get silenced out. But many of the breaths that come before a phrase get left in, since people are used to hearing people breathe before they talk.
One more warning – if you silence most of the space between phrases, the start of a pre-phrase breath may be cut off. THAT will make the breath sound unnatural. You can fix that problem either by making sure the section you select for silencing does not include the start of the breath. Or you can use the "fade-in" tool if the start of a breath gets cut off. Creating a fade-in makes it sound natural again.
I hope this helps you get much better-sounding voice recordings!
By the way, the Audio Daily Blog post I referred to at the beginning of this article is here: http://blogs.voices.com/audiodaily/2013/05/6_tips_for_removing_breaths.html


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The Dot Line DLC Manual Extension Tube Set allows the lens to be moved away from the camera for ultra close-up photography. The camera must be used on the manual mode. Just attach your lens and move the camera in until the object is in focus.
|Tube sizes:
|12, 20 and 36mm


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Hotels near Bel Air
Browse 21 attractions, meet 8 travelers, 2 tour guides and discover 29 photos
Travelers are voting Best Budget Inn Havre De Grace, Clarion Hotel Aberdeen and Esa Baltimore-timonium as the best of 124 hotels near Bel Air. There are 181 hotels in New Market a city just 109 km from Bel Air and 269 hotels in Rockville which is 108 km distant. The most popular of these are Express Inn Knoxville in New Market and American Inn in Rockville. You can find 592 hotels in Maryland and 44568 hotels in the country of United States. Our members favorite in Maryland is Best Budget Inn Havre De Grace and Pelican Hill is the top pick in United States. Want more? Check out our map of hotels near Bel Air.
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The emergence of blood abnormalities, an adult ability to digest milk, and changes in our physical appearance point to the continued evolution of the human race.
Infographic: Advancing Forensic Science
January 1, 2017|
Forensic scientists have been using rudimentary molecular techniques for decades. But advanced forensic anthropology technologies and methods are just now coming to the fore in some investigations.


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Matt Shoemaker to make 2016 debut for Angels on Friday night
ANAHEIM -- Matt Shoemaker, who is seeking to prove that 2015 was the aberration and 2014 was the real thing, will officially begin that process with his first start of the season Friday night against the Texas Rangers.
Shoemaker was a revelation in 2014, when he was 16-4 with a 3.04 ERA and he finished second in Rookie of the Year voting. Last year, though, he slumped to a 7-10 record and a 4.46 ERA. He was briefly sent to Triple-A.
Shoemaker might be facing the right team. He is 4-0 with a 2.39 ERA in his career against Texas.
The Rangers will counter with A.J. Griffin, who also has something to prove. Griffin has missed the last two seasons with injuries. He had Tommy John surgery and then shoulder trouble. Griffin was once one of the Oakland A's most promising young pitchers.
The Angels lineup has a couple interesting changes for the game. C.J. Cron and Johnny Giavotella both have the night off, with Ji-Man Choi and Cliff Pennington getting their first starts of the season, at first and second, respectively.
The lineups ...
Yunel Escobar 3B
Daniel Nava LF
Mike Trout CF
Kole Calhoun RF
Andrelton Simmons SS
Ji-Man Choi 1B
Cliff Pennington 2B
Delino DeShields CF
Shin-Soo Choo RF
Prince Fielder DH
Adrian Beltre 3B
Mitch Moreland 1B
Ian Desmond LF
Roughned Odor 2B
Elvis Andrus SS
Robinson Chirinos C
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Each profile within the State Profiles reflects SIECUS' focus on the nexus of policy and implementation, providing a portrait of sexuality education and abstinence-only-until-marriage programs in the United States. This 11th edition includes information from federal fiscal year (FY) 2013, October 1, 2012 – September 30, 2013.
View the Puerto Rico State Profile At A Glance
This overview highlights the state of adolescent sexual health promotion efforts in Puerto Rico for FY 2013 including sexuality education laws and policy, sexual health data, and federal funding.
View the FULL Puerto Rico State Profile
This FY 2013 profile for Puerto Rico includes information (where applicable) on sexuality education laws, policies, and legislation; youth sexual health and behavior data; and federally funded programs as implemented in the territory, with relevant contact information.


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FE170: Building Confidence For All Sports
Instructor: Amanda Nelson
Course Details
Does your dog lack confidence when competing? Or going out for walks?
Maybe they get nervous when company comes over?
In this class, we will be working on different games and mental activities to help boost your dog's self-esteem and confidence. whether it is just in daily life or competing.
We will play a lot of mental games that will focus on boosting the dog's confidence and self-esteem, letting the dog "come out of their shell".
This class will follow along with my dog Ally through the training videos as she builds her confidence with these games as well!
Games will include; targeting, mat work, cone work, clicker games, hide and seek games, and more!
There are no scheduled sessions for this class at this time. We update our schedule frequently, so please subscribe to our mailing list for notifications.
Registration will begin at 10:00AM PDT.
Enrollment limits: 15 gold, 25 silver, unlimited bronze. If you are interested in a bronze-level spot you can register any time during the registration period.
Week 1:
What is Confidence?
What is Self Esteem?
Food vs. Toy
Week 2:
Build the Click
Fun with Targets
Week 3:
Cones, Cones, everywhere!
Can you find it?
Week 4:
Ready?! Go!!
Week 5:
Ready?! Wait!!
Week 6:
Advanced Target and clicker games
Prerequisites and Equipment
Bath Mat (or rug)
Road cone (anything the dog can do around. Chair, garbage can, etc.)
Dixie cups (paper cups,red solo cups, or even plastic bowls)
Items to target ( flower pots, boxes, plates, etc. We will be targeting to "different" items as well as traditional)


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Reviews and Notes: Analyzing How We Reach Clinical Decisions. Ann Intern Med. 1994;121:155. doi: 10.7326/0003-4819-121-2-199407150-00032
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Published: Ann Intern Med. 1994;121(2):155.
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I am using nDVR addon and I want to achieve adaptive bit rate streaming. My encoder is capable to creating key frame aligned streams from a single stream. I have modified the nDVR HTTPProvider code (com.wowza.wms.plugin.dvrstreamrecord.HTTPDvrStreamRecord) to start and stop recording of the multiple streams by calling following url.
The modification that I did just calls "start record" function on the stream names inferred from "streamnameprefix" parameter. So lets say I have 3 stream then it will call "start record" function 3 times on each stream one after another. Is there a different way to start multiple stream recordings simultaneously.http://[wowza-ip-address]:8086/dvrstreamrecord?app=[application-name]&streamnameprefix=[stream-name-prefix]&recordingname=[recording-name]&action=[start|stop]


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We conclude "Why? Week" with a Glorious Post about The Most...Wonderful Time...of the Year!
A little background...The Scribe Family once lived on Capitol Hill in DC, in the Eastern Market neighborhood. Eastern Market, BTW, is the longest-running open-air market in the Nation's Capital. Its more than 100-year-old brick centerpiece burned down last year. The neighborhood rallied to save this piece of history, & the Market is now on its way to being Good As New; or actually, Good As Old, since the area is in the National Registry of Historic Places.
We lived on The Hill for 12 years, in a beautiful 19th-Century restored (not by us...we're not handy!) rowhouse, circa 1837. As a side note, the garden was so fertile we grew tomatoes that measured up to 8 inches in diameter! You see, Back in the Day, when horses boarded out back, and folks just chucked their garbage in the yard...
Anyway, back to this Sweet Snap. It's actually a Polaroid, which has hung on the tree as an ornament for 16 years, give or take 1 or 2. That many years creates Damaged Goods. So Mrs. Scribe took the matter into her own hands this year, and scanned and PhotoShopped the sucker. It's still not perfect, but we think you can see the pure joy & amazement on the faces of these two chicas. Meet Ella Numera Una & her BFF. They're actually Sitting. On. Santa's. Lap!
Santa has been coming to Eastern Market for years. He always arrives on a Sunday, in a horse-drawn carriage. We're gonna take this snap to a professional, to have it restored correctly. But on the Friday before Christmas, we wanted to Wish One & All Good Tidings!


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With Instructables you can share what you make with the world, and tap into an ever-growing community of creative experts.
Tell us about yourself!
Hello there, I found your video on YouTube recently and tried to build one of those myself. The results are not so good as yours, but that's my first attempt to build anything from coca cola cans. Here's what I've got right now:
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You purchased your SX 2.0 because its reliable transport and versatility struck a chord inside you; so why would you settle for second rate when it comes to superior quality auto parts? Anyone who operates a Dodge knows that when it comes to repairing their automobile only the highest quality replacement parts will do. It's not every day that you discover you need a replacement Brake Rotors & Discs to fix your automobile. Obviously, you didn't buy your SX 2.0 unless you understood you would be getting great performance and great value both impeccably combined into one vehicle.
The ability to slow down and make quick evasive maneuvers is crucial to keeping you and your passengers safe. Rather than being spread apart like old brake drums were, disc brake rotors get clamped in one spot by brake pads. This means Dodge SX 2.0 disc brake rotors take a beating from your brake pads, but they are made from a hard metal so they don't wear away like the pads do. Brake rotors perform very well in rain (much better than drums), and are virtually always included from the factory on current vehicles. Your Dodge SX 2.0 owes its ability to slow down thanks to the brake rotors bolted to the wheel hubs.
When your Dodge needs a new replacement Brake Rotors & Discs, there's no need to waste time looking anywhere other than the authority, Car Parts Discount. Ordering new Dodge parts from www.carpartsdiscount.com means you are getting the best deal on the absolute greatest original and aftermarket replacement SX 2.0 parts on the internet. Trust in the value of the parts you get from carpartsdiscount.com and save time and money on all your upcoming repairs. All the Dodge SX 2.0 Brake Rotors & Discs parts on this page will fit vehicles made in 2005, 2004, 2003. Choose your year below to refine your search.


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Take away.
Take away.
We went out for Indian we got
Butter chicken
Beef vindaloo
Naan bread & popadoms
Carpe diem 2013 (22-08-2014)
Thai take out, yum.
ImageUploadedByThe Bub Hub1408706448.475565.jpg this is what hubby just served me up for dinner. Trying to drop a hint perhaps..?..!..
bunkx (22-08-2014),Carpe diem 2013 (22-08-2014),Teenz (23-08-2014),~Marigold~ (23-08-2014)
Mushroom and bacon risotto thanks to hubby. Vanilla custard for dessert.
Fresh fish and chippies
♥ me & my little boy ♥
Steak & potato bake
DH + Me + DD (born 11/10/12) = our little family
Steak & potato bake, and veg
Pregnant for the first-time?
Not sure where to start? We can help!
Our Insider Programs for pregnancy first-timers will lead you step-by-step through the 14 Pregnancy Must Dos!


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On the heels of his neon-infused visuals, Italian designer Marcelo Burlon adds to the support of his Fall/Winter 2013 T-shirt line with his "Country of Milan" video. Drawing iconographic elements from different cultures – including Mapuches crosses from Patagonia, Argentinean birds feathers and more Gothic themes often associated with people like Riccardo Tisci of Givenchy, the result is a more straightforward unveiling of the various items than past visual offerings. Expect to see the entire range of tees at select stockists around the world in the near future.


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Standard Single Room - With Breakfast
- Breakfast
Price for 1 Night
Rs. 2,653Rs. 2,293
2 Rooms Left!
Room Overview
Standard Room
- Breakfast
Fri, 03 Mar – Sat, 04 Mar (1 Night) | 1 Room | 1 Guest
Opp. Japanese Temple, Bodhgaya , 824231 | Show on map
Save Rs. 360
Rs. 2,293
Standard Room
Standard Room
I alongwith my wife reached to BodhGaya, HotelTokyo Vihar at about 10.45 night on 27.04.2012.The receiption official was very keen and after seeing our hotel voucher he immediately given us a double occupancy , executive /ac room having balcony in the side. The hotel location is very good just in front of Japanese Temple so the name of the Hotel.Even at that night time they provided us tea and light refreshments on our request. The Parking in the hotel area and outside is sufficient for the occupiers.The reception lobby and the dinning hall are very spacious and neat and clean. Qualities of foods , snacks etc. is excellent and well prepared and to the reasonable cost.Parking and location of the hotel is excellent i.e very near to the main Buddha temple and other Buddy's temples of various countries.The front desk staff is very cooperative. Further the room were good, neat and clean, Bathroom are quite spacious. In the end overall we can say that we stayed in the hotel from 27.04.2012 to 01.05.2012 and felt homely pleasure in food quality, room cleaniess and receiption and dinning halls. However in every structure/hotel there are always room for the improvement.
(based on 65 reviews)
by markaclake on February 24, 2017
Service was good and the food was awesome, it was very comfortable to get the famous temples nearby in a walk in distance from the hotel. Rooms were good clean. Staffs were ok. The area is quite noisy at night so we didn't sleep too well
by Chiara M on January 22, 2017
The service was very good, they received us with a cold drink and our room was clean, plus the bathroom had an actual bath tub in it, which meant that we didn't have to flood the floor when taking a shower. The restaurant food was not good though, and the wifi didn't work from our room but only from the reception hall. The area is quite noisy at night so we didn't sleep too well.
by DANNY T on December 24, 2016
I booked a room for 2 nights and the hotel was very good and the staffs are excellent. Service was good and the food was awesome, it was very comfortable to get the famous temples nearby in a walk in distance from the hotel. Overall a very pleasant stay
by Zelpi on August 12, 2016
Arrived late in Gaya and asked for a room. The hotel per se was not that bad, room was fine, no towels. But the staff was really rude towards us as if we would have been undesired guest, reminding us more than once about checkout timings. Stood one night then left this place!!
by Darren C on July 7, 2016
I booked a room for two nights the managers were very unfriendly, my air con didn't work so after lots of complaints I got a new room that wasn't much better. The power kept going out and the air con was on and off all night, the room phones didn't work as they weren't hooked up, some of the power switches sparked when turned on and the managers didn't seem to care. You get no wifi past the first floor so I was using it in the lobby mad at 11pm I was told I had to stop using wifi a go back to my room as the reception was closing, the upon going to reception in the morning the manager was sleeping on the lounge so I don't see why I could not sit in there myself. Over all rude staff and a poor stay at this hotel, I would not stay here again and would not recommend this place.
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by DANNY T on December 24, 2016
by sanjay744 on May 6, 2016
by Prasannjeet on March 4, 2016
by Mo_dhawan on September 29, 2015
by soumohtv1 on August 2, 2015
Cancellation and prepayment policies vary according to room type. Please check what room conditions may apply when selecting your room above.


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This product is available. Order NOW!
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|Moisturizes and protects from pollution and metals (Nickel, Chromium, Cobalt...)
|Recommended for
|Body and/or face
|13 ingredients : Aqua (Water), Isononyl Isononanoate, Calcium Carbonate, Ricinus Communis (Castor) Seed Oil, Cetearyl Alcohol, Glycerin, Glyceryl Stearate SE, 1,2-Hexanediol, Ceteareth-20, Carbomer, Chlorphenesin, Tocopheryl Acetate, Xanthan Gum
|2 pump dispensers of 6.76 Fl oz / 200 ml
- Normal skin
- Dry skin
- Sensitive or reactive skin
- Metal allergy (Nickel, Chromium, Cobalt…)
- Eczema or dermatitis
- Dyshidrosis
A very pure moisturizing body lotion which hydrates and protects from pollution and metals (Nickel, Chromium, Cobalt…).
Exclusive patented technology. For dry, sensitive or allergic skin (Nickel, Chromium, Cobalt…).
- Dry skin
- Sensitive, reactive skin
- Nickel allergy
- Chromium allergy
- Cobalt allergy
- Eczema, dermatitis
- Dyshidrosis
Exclusive patented technology
The Moisturizing Lotion HP is based on the exclusive patented Capturifique technology. This technology captures metals (Nickel, Chromium, Cobalt…) at the surface of the skin.
Some of this cream's ingredients are assembled in specific structures. These structures capture the metal irritants through a chelation mechanism.
The structures also have physicochemical properties that make them stay on the surface of the skin. Therefore, when metals come in contact with the skin, they are captured and prevented from entering (or re-entering) the epidermis.
The metals can then be washed away from the surface of the skin by cleaning it with the Cleanser P, or with water.
Learn more about the Capturifique technology
Excellent skin comfort
This lotion has a fluid, fast-absorbing and non-greasy texture. It lets the skin breathe naturally, without leaving a barrier film.
Only 13 ingredients to minimize the risk of skin reaction. Each ingredient has been carefully selected for its safety profile.
- fragrance-free
- paraben-free
- silicone (dimethicone) free
- sulphates free
- phthalates free
- mineral oil free
- lanolin free
- EDTA free
- Nickel, Chromium, Cobalt free
Hypoallergenic, tolerance tested.
Environmental friendly and not tested on animals.
Results of the scientifically controlled tests:
- 35% moisture increase in just one day
- 50% moisture increase over longer periods (applied on the body once a day)
Apply generously on dry skin and massage until complete penetration of the product.
Use daily and as frequently as necessary.
3 rating(s)


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In a daring incident of road rage five students, including one of Lovely Professional University (LPU), Jalandhar, beat up the driver of a Mahindra Bolero pick-up van of a private company and snatched away the vehicle after the van had hit their car on the road dividing sectors 7 and 8, here on Wednesday evening. Later, the police arrested two of the youths for dacoity and recovered the snatched vehicle.
Giving details, sources in the police said five students were travelling in a car around 7pm when a Bolero van hit the car in Sector 7. The students chased the Bolero and stopped at the intersection dividing sectors 7, 8,18 and 19 on Madhya Marg. Sources said two students have been identified as Jasnit, pursuing MSc from Karnataka University, and Manpreet, a student of civil engineering at LPU. Their three friends are yet to be identified.
The pick-up van was being driven by Tarsem Lal, a resident of Bapu Dham Colony. He said the youths, soon after stopping him in the middle of the road, thrashed him badly. After that, Jasnit and Manpreet snatched the keys of the van and drove away and their other friends left the place in the car.
Tarsem Lal raised an alarm, following which, police were informed and the vehicle number was flashed. The pick-up van was finally intercepted at the Sector 47/48 barrier. The police later arrested two students and registered a case of dacoity against them at the Sector 26 police station.
ASP Parminder Singh said the medical examination of the youths was being conducted and it seemed that one of them was drunk. "Efforts are on to nab the other three accused," he said.


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A word game for iOS where you move tiles around to create words.
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Jungmaven Women's Baja 100% Hemp Tee
Jungmaven Women's Baja 100% Hemp Tee
Sustainably-made, perfectly fitted, everyday tee in 100% hemp. Available in urban black.
Shipping and returns (when eligible) are FREE for purchases from US and Canadian boutiques when shipped to customers located in the US or Canada. We offer a flat shipping fee of $30 for all international transactions.
Shipping Details
Returns are accepted on full-priced items except when item is marked as Final Sale. All sale items, including Make an Offer items, are FINAL SALE.
Returns Policy
A pioneer of Seattle's independent designer boutique scene, Velouria has set up shop in the up and coming Pioneer Square neighborhood. With white walls and a gallery feel, it's the perfect spot for their ever-changing and vast selection of small production, independent and local designer wares. You'll find designers such as Dusen Dusen, Feral Childe, Jennifer Glasgow and more.


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TipTopHaus by Josh Wynne Construction
This home is constructed in the world famous neighborhood of Lido Shores in Sarasota, Florida. It features a flipped layout with a front court pool and a rear loading garage.
Ryan Gamma Photography
- Modern Bachelor by Contour Interior Design
- Cowrie Road Home by MG Design & Building
- White House by In2
- Fisher Island by Associated Design Co
- Marin Residence by Dawson & Clinton
- Cozy Home by Ruetemple
- Loft in San Diego by Hawkins + Hawkins Architects
- Cabin Norderhov by Atelier Oslo
- House Hohlen by Jochen Specht
- Le 205 by Atelier Moderno


text: Dtp Operator Port Blair. Job Openings in Mohan Impressions for Dtp Operator Port Blair, this job opened at 21 Feb, 2017. To design various type of templates through Photoshop and Corel Draw. _Salary: _ INR 60,000 - 1,25,000 P.A. _Industry: _ Printing / Packaging _Functional ... (Read more about Dtp Operator Port Blair - Port Blair - Mohan Impressions)