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2NE1 CL Wears a Mini Dress and Reveals her Sexy Body
By Staff Writer | July 31, 2013 07:53 AM EDT
Today she posted on her instagram, "Do you love me?" along with a picture.
In the picture, use is showing off a sexy pose in a black mini dress.
Internet users commented, "She is getting sexier" and "She looks so good in black."
On the other hand, 2NE1 is busy with their new song, "Falling in Love."
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The Arizona-based "Those Guys" team didn't come away with the championship at the Las Vegas NLA 7-on-7 over the weekend, but for tight end Trevor Wood (Scottsdale, Ariz./Chaparral), just getting out on a football field was victory enough.
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Autotune. U can take a fairly attractive face with a willingness to shock and assign them a "sound". I have gone above and beyond to teach my children the difference between art and the money making propaganda machine. Some of this stuff has a catchy beat and people call it music. There is very little if any individuality in todays media. Movies are included. Look at all of the cgi remakes. They are just out of ideas. And it is a business. Lets face it... It is easier to find some kid willing to sell his soul for fame with no desire in the creative process. As long as they get fame, money and sex. That is much easier than actually producing quality media.
Janine u r much too pretty and young looking to have experienced those bands haha. I mean i would not have guessed it by looking at ur pic but good for you! Those are memories that are becoming harder to come by. Glad you got the opportunity to see such classics. I have an extensive list of my own.
The last concert I attended was last October just 5 miles from my house. I saw Gary Wright and Al Stewart at the Silver Reef Casino. They are old yes but...they LOVE to perform! I got to meet them after. I saw Peter Gabriel a few years ago.
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Mr. Nakazawa and Yamazawa equally contributed to this study.
Research Article
Formation of the digestive tract in Ciona intestinalis includes two distinct morphogenic processes between its anterior and posterior parts
Version of Record online: 29 JUL 2013
DOI: 10.1002/dvdy.24009
Copyright © 2013 Wiley Periodicals, Inc.
Developmental Dynamics
Volume 242, Issue 10, pages 1172–1183, October 2013
Additional Information
How to Cite
Nakazawa, K., Yamazawa, T., Moriyama, Y., Ogura, Y., Kawai, N., Sasakura, Y. and Saiga, H. (2013), Formation of the digestive tract in Ciona intestinalis includes two distinct morphogenic processes between its anterior and posterior parts. Dev. Dyn., 242: 1172–1183. doi: 10.1002/dvdy.24009
Publication History
- Issue online: 20 SEP 2013
- Version of Record online: 29 JUL 2013
- Accepted manuscript online: 29 JUN 2013 12:00AM EST
- Manuscript Accepted: 17 MAY 2013
- Manuscript Revised: 2 MAY 2013
- Manuscript Received: 12 JAN 2013
Funded by
- JSPS. Grant Numbers: 18370088, 22570207
- Abstract
- Article
- References
- Cited By
- Digestive tract morphogenesis;
- Ascidian;
- Ciona intestinalis;
- Endodermal strand;
- Endoderm
Background: In the ascidian Ciona intestinalis, the digestive tract, an essential system for animals, develops during metamorphosis from the two primordial tissues, the endoderm and endodermal strand, located in the larval trunk and tail, respectively. However, it has been largely unknown how the digestive tract develops from these primordial tissues. We examined the metamorphosing larvae for the tubular formation of the digestive tract, focusing on the epithelial organization of the endoderm, by combined confocal microscopy and computational rendering. Results: The tubular structure of the esophagus to the stomach was formed through the folding and closure of the endodermal epithelia in the central-to-right posterior trunk. By contrast, the intestine was formed in the left posterior trunk through the accumulation and rearrangement of the cells originated from the endodermal strand. This was confirmed by the cell-tracing experiment using Kaede expression construct driven in the endodermal strand. Thus, the tubular formation of the digestive tract in C. intestinalis includes distinct morphogenetic processes and cell lineages between its anterior and posterior parts. Conclusion: This study provides the first detailed description of the digestive tract morphogenesis in C. intestinalis and serves as an important basis toward thorough understanding of its digestive tract development. Developmental Dynamics, 242:1172–1183, 2013. © 2013 Wiley Periodicals, Inc.
The digestive tract is an essential system for animals, and it develops from the endoderm. In that development, several processes such as endoderm formation, endoderm patterning, duct formation, and organ specification must be included. To date, molecular analyses of each of these processes as well as the morphogenetic descriptions of the digestive tract in chordates have been reported largely on vertebrate embryos (Zorn and Wells, 2009). In vertebrate embryos, however, it is rather difficult to analyze thoroughly the process of their digestive tract development due to the anticipated complexities. By contrast, such studies may be feasible in primitive chordate ascidians with simple body plans of their larvae and adults.
Ascidians belonging to the subphylum Urochordata are regarded to be the closest extant relative of vertebrates (Delsuc et al., 2006; Putnam et al., 2008). Ascidian eggs develop into tadpole larvae, which share the prototypical morphogenesis and body plan of chordates characterized by the formation of a hollow dorsal neural tube, notochord and paraxial mesoderm (Satoh, 1994, 2003). Nevertheless, larvae in most ascidian species, including Ciona intestinalis, do not possess the digestive tract. Instead, the larva possesses the digestive tract primordia in two regions: one in the trunk (the head of the tadpole-shaped ascidian larva) and the other in the tail. They are designated the endoderm and the endodermal strand, respectively. Formation of the digestive tract takes place during the metamorphosis, when the tail is absorbed into the trunk (Satoh, 1994). The endoderm is the tissue, consisting of approximately 500 undifferentiated endodermal cells, and is placed in the ventral trunk containing a luminal space designated "the endodermal cavity" located in the middle of the tissue (reference therein, Satoh, 1994). It has been shown with the metamorphosing larva of Halocynthia roretzi that the anterior endoderm forms the branchial basket consisting of the branchial sac, endostyle, peripharyngeal band, dorsal tubercle, and peribranchial epithelium, and the posterior endoderm develops into the digestive tract, consisting of the esophagus, stomach, and intestine (Hirano and Nishida, 2000). However, the detailed information regarding the ways the digestive tract is formed and/or which digestive organ develops from which part(s) of the posterior endoderm is unknown. This holds true with the endodermal strand that is the tissue, running ventrally along the length of the notochord in the tail (Satoh, 1994). It is unknown how the endodermal strand is involved in the digestive tract formation.
In the ascidian, C. intestinalis, juveniles have a relatively large branchial basket and a rather simple digestive tract, consisting of four digestive organs, the esophagus, stomach, intestine, and the pyloric gland being the only accessory gland (Chiba et al., 2004). Although the digestive tract of C. intestinalis is relatively simple, our previous study revealed that Hox genes, Hox10, Hox12, and Hox13, are expressed in the posterior part of the digestive tract with distinct expression domains aligned along the anterior–posterior axis (Ikuta et al., 2004). This is reminiscent of the developing hindgut in chicken embryo, where Hox10, Hox11, and Hox13 genes are expressed and their individual expression domains are aligned along the A–P axis in the gut's endoderm (Yokouchi et al., 1995). These observations suggest that some molecular mechanism of regionalization of the digestive tract may be shared between the ascidians and vertebrates. Furthermore, several of transgenic lines, which harbor green fluorescent fluid gene expressed in the specific region within the digestive tract of the juvenile, has been isolated in C. intestinalis (Yoshida and Sasakura, 2012). Thus, C. intestinalis, together with its abundant genomic as well as cell lineage information, is an attractive model for studying the digestive tract development and its diversity in chordates.
In this study, we have first described the morphogenetic process of the digestive tract, because it has been largely unknown how the tubular structure of the digestive tract develops either in C. intestinalis or in other ascidian species. For this, the metamorphosing larvae and juveniles were fixed and stained with phalloidin, and optical sections were obtained sequentially using a confocal laser-scanning microscope. Focusing on the epithelial structure, the formation of the tubular structure of the digestive tract was traced four-dimensionally. We found that the endodermal cells located posterior central to right of the trunk width, facing the endodermal cavity, became epithelialized and formed the esophagus and stomach by folding and closure of the endodermal epithelia. By contrast, the intestine was formed in the central to left of the trunk width through accumulation and rearrangement of the cuboidal cells largely of the endodermal strand origin. This was confirmed by a novel cell lineage-tracing experiment, using a Kaede expression construct. The present study provides the first detailed description of the tubular structure formation in the digestive tract development in C. intestinalis.
Computational Rendering Reconstruction of Outlines of the Digestive Tract in the Juvenile After Body Axis Rotation
To describe tubular structure formation processes of the digestive tract, we used confocal microscopy with phalloidin staining to recognize the epithelial tissue, the most conspicuous structure of the digestive tract, and rendering procedure to construct three-dimensional (3D) models out of the epithelial tissue images of developing digestive tract.
As the first step, we examined the inner structure of C. intestinalis juveniles at stage 37, the late body axis rotation stage, since Chiba et al. have reported that the digestive organs are recognizable at this stage (Chiba et al., 2004). As shown in Figure 1, a tubular epithelial structure was clearly recognizable within the juvenile body (Fig. 1A–F). From the left to central of the juvenile body width, the intestine was located spanning along the body axis. The posterior end (the anus) of the intestine was located close to the left atrial siphon primordium (Fig. 1A,B), and the other end was opened to the large lumen of the stomach, occupying central to right of the juvenile body width (Fig. 1C–E). The stomach was, in turn, connected to the esophagus on the right side (Fig. 1D–F). The opening of the esophagus to the branchial basket was seen above the bulge of the stomach (asterisk in Fig. 1D), thus the esophageal duct was found to be located in the central to right of the juvenile body width. The digestive tract, spanning from the esophagus to the end of the intestine, is schematically shown in Figure 1G. Additionally, the pyloric gland, an organ branching from the posterior stomach and lying along the intestine, was identifiable (Fig. 1B,C). On each cross-sectional image of the digestive tract, epithelium was colored manually on computational images, and with the images of colored epithelia, a 3D model of the digestive tract was constructed using DeltaViewer software (see the Experimental Procedures section). The reconstructed 3D model clearly showed the outlines of the digestive tract of a metamorphosing juvenile at stage 37 (Fig. 1H–J), suggesting this rendering method to be useful.
Reconstruction of Outlines of the Digestive Tract in the Juvenile During Body Axis Rotation
Next, we examined the larvae during the body axis rotation (stages 35 to 37) for the development of the digestive tract epithelia (Fig. 2A–D), and 3D models were constructed (Fig. 2E–L). The images showed that the digestive tract at stage 35 had a very similar architecture, but consisted of a simple smaller tube in comparison to that at stage 37 (Fig. 2E,I,H,L). In other words, we suggest that it can be seen that the simple small tube is enlarged during body axis rotation stages, and forms the digestive organs (Fig. 2E–L). This formation is accompanied by the distinct morphological changes, such as expansion of the stomach and elongation of the esophagus and the intestine. The pyloric gland becomes recognizable on the outer surface of the intestine at early stage 36 (Fig. 2J) and is elongated along with the intestine, keeping the end of the gland at the central intestine (Fig. 2K,L).
A series of sections of the 3D model at the levels shown in Figure 2M revealed a tubular lumen of the digestive tract at stage 35 (Fig. 2N), which starts dorsally (b in Fig. 2N), goes rightward, turns toward the ventral (a in Fig. 2N), then goes leftward and finally turns toward the dorsal (f in Fig. 2N). The sections of the 3D model also indicate the presence of a branching lumen, corresponding to the pyloric gland (d, e in Fig. 2N).
Considering the above observations, it is suggested that the esophagus, stomach, and intestine are formed in the right, center, and left regions of the posterior trunk, respectively, before the onset of body axis rotation. Therefore, in the following sections, the three regions of the metamorphosing larvae were separately examined to determine how and when the tubular lumen of the digestive tract formation takes place.
Duct Formation of the Prospective Stomach in the Central Posterior Trunk of the Metamorphosing Larvae
In the larva at early stage 27, immediately after hatching, we examined sagittal sections (Fig. 3). This revealed that only a short stretch of epithelium was present in the dorsal endoderm, while ventral endodermal cells were not epithelialized yet (Fig. 3A). By contrast, in larvae at late stage 27 onward, both ventral and dorsal endodermal cells facing the endodermal cavity were completely epithelialized (Fig. 3B–F). Three-dimensional models rendered with the epithelia clearly showed duct-forming processes in the central region (Fig. 3G–K). At late stage 27, the epithelium facing the posterior endodermal cavity constituted a V-shaped furrow along the left–right (L-R) axis (Fig. 3G). At stage 28, the anterior–dorsal and anterior–ventral edges of the V-shaped epithelial furrow came close to each other (blue arrows in Fig. 3H) and eventually fused by stage 29 (Fig. 3I), which resulted in a small luminal space (red lining in Fig. 3I). At stage 30 onward, the cells surrounding the luminal space became columnar (Fig. 3E) and the luminal space-surrounding cells were completely separated from the endodermal cavity epithelium (Fig. 3J,K). Thus, a duct running along the L-R axis was formed ventrally through folding and closure of the epithelia in the posterior central trunk by stage 32 (Fig. 3K).
In addition, the 3D models viewed from the opposite side (Fig. 3M–O) unveiled that the lumen of the duct became divided into two as development proceeded. One duct located ventrally would form the main digestive tract and the other located dorsally would form the duct of pyloric gland.
Tubular Lumen Formation of the Prospective Pharynx and Esophagus in the Right Posterior Trunk of the Metamorphosing Larvae
Tubular lumen formation in the right posterior trunk of the larvae at stages 29, 30, and 32 was examined at three parasagittal levels (a, b, and c in Fig. 4A) by constructing 3D models out of the endodermal epithelia (Fig. 4B–I). In the sections at the level a, it was revealed that the tubular lumen formation became evident in the ventral endodermal epithelium at stage 29, and additionally, another tubular lumen formation took place dorsally around stage 30 (Fig. 4B–D). The former tubular lumen formation was very similar to that described in the previous section. The latter tubular lumen formation was evident at the level b (Fig. 4E,F) but it was incomplete at the level a, leaving the ventral side of the lumen open to the endodermal cavity (Fig. 4D). At the level c, it was revealed that the luminal space formation was initiated in a way reminiscent of that in the central region. However, this resulted in an elongated luminal space along the D–V axis (Fig. 4G–I). The luminal space eventually connected the two tubular lumens running dorsally and ventrally along the L-R axis (Fig. 4I,J). The dorsally running tubular lumen opened to the endodermal cavity at the immediate right to the notochord (Fig. 4D,J). We suggest the tubular lumen and the opening correspond to the esophagus and its anterior end connecting to the pharynx, and the prospective esophagus was connected to the prospective stomach located ventrally (Fig. 4I,J). Furthermore, the distorted tubular lumen of the prospective stomach was divided into two tubular lumens, continuing to the prospective intestine ventrally and to the pyloric gland dorsally (Fig. 4J, see also Fig. 3O). Thus, the anterior prospective digestive tract, from the pharynx to the stomach, was formed through folding and closure of the endodermal epithelia by stage 32 (Fig. 4J).
Prospective Intestine Formation in the Left Posterior Trunk of the Metamorphosing Larvae
In the next stage, posterior left trunk in larvae at stages 27 to 32 was examined for the development of the intestine. Because the intestine in the juvenile connects the stomach and the opening to the left atrial siphon, first, parasagittal sections at the level immediate right to the atrial siphon primordium were examined (Fig. 5).
In larvae at stage 27, epithelium-like cells were observed in the region posterior right to the atrial siphon primordium, where the prospective intestine develops (Fig. 5A). However, epithelium-like cells were missing from the position at stage 28 (Fig. 5B). By contrast, in larvae at stage 29, a cluster of cells with epithelium-like appearance consisting of large cuboidal cells was emerging in the region posterior right to the atrial siphon primordium and dorsal to the prospective stomach, or the duct to be connected to the prospective stomach (Fig. 5C). This cluster changed its contour shape as the development proceeded (Fig. 5D–F). In larvae at stage 30, a lumen was formed, being surrounded by most, but not all, of the cuboidal cells (upper red lining in Fig. 5D) on the dorsal side of the duct connected to the prospective stomach (Fig. 5D). In larvae at stage 32, a tubular lumen running along the D–V axis was observed (Fig. 5F). The lumen is expected to be of the prospective intestine (Fig. 5F). The anus did not open to the atrial siphon until juvenile stage (data not shown). Thus, the findings out of the parasagittal sections suggest that the intestine forms through accumulation and rearrangement of the large cuboidal cells.
Cells Forming the Prospective Intestine Originate From the Endodermal Strand
To explore the origin of the cuboidal cells, horizontal sections at the level ventral to the notochord were examined in larvae at stages 27 to 32 (Fig. 5A–F,G–L). In the larva at stage 32, the duct of the prospective intestine was located on the left side of the prospective stomach (Fig. 5L). By contrast, in the larva at stage 31, the duct was not so evident but a group of large cuboidal cells was recognizable in the prospective intestine region (Fig. 5K). In the larvae at stages 29 and 30, the large cuboidal cells were aligned in a row spanning from the prospective intestine forming region to the more posterior central region (Fig. 5I,J). In the larvae at stages 27 and 28, the prospective intestine cells were hardly identifiable among cells of similar morphology, and neither were the large cuboidal cells in a row (Fig. 5G,H). In the sagittal section of the larva at stage 29, a stretch of cells with somewhat similar morphology to the cuboidal cells was observed aligned along the ventral to the anterior notochord (Fig. 5N). These cells are aligned as continuation of the endodermal strand cells that are located as a file of squamous cells immediate ventral side of the notochord in the tail (Fig. 5N). These cells and/or the endodermal strand cells were missing from the posterior to anterior as development proceeded (Fig. 5N,O) and disappeared by stage 31 (Fig. 5P).
Based on these observations, we hypothesized that the cells of the endodermal strand move toward anterior into the left posterior trunk of the larva, where they form the intestine. To test this hypothesis, we labeled and traced the endodermal strand cells using the larvae introduced with pZip-Kaede expression construct and those of the transgenic line harboring the pZip-Kaede expression construct (see the Experimental Procedures section), which drives the expression of the Kaede protein in a part of the sensory vesicle, posterior endoderm and in approximately anterior two-thirds of the endodermal strand at larva stage (Fig. 6A, see the Discussion section).
A larva at stage 27 introduced with the pZip-Kaede expression construct, emitting green but not red fluorescence of the Kaede protein in the endodermal strand (Fig. 6A, data not shown), was irradiated with ultraviolet (UV) light only at anterior two-thirds of the tail. Immediately after UV irradiation, it was confirmed that the larva emitted the red fluorescence of the converted Kaede protein solely in the endodermal strand (Fig. 6B,C; red fluorescence is represented by magenta color). Such larvae were allowed to develop and were examined for the red fluorescence at various stages. In larvae at stage 30, well before the tail absorption, red fluorescence was located in the left posterior trunk, but not in the tail (Fig. 6D,E). It should be noted that left-sided localization of the red fluorescence was always the case, so far as the larvae of the stable pZip-Kaede transgenic line developed within the chorion were concerned (n = 21). In the larvae introduced with pZip-Kaede by electroporation, the left-sided fluorescence localization was approximately one-quarter (n = 11), probably due to the removal of the chorion as has been reported with the left-sided expression of Ci-Pitx and Ci-Nodal (Yoshida and Saiga, 2008, 2011). When the larvae metamorphosed into juveniles, red fluorescence was detected in the intestine (Fig. 6G) and some other parts, including the periesophageal body and the tail residue (Fig. 6G′, see the Discussion section). These results indicated that the endodermal strand cells move from the tail into the left posterior trunk, which occurs well before the tail absorption, and they contribute to form most part of the intestine.
In the present study, we have described for the first time the morphogenetic processes of the digestive tract in the ascidian, C. intestinalis on the basis of confocal microscopic observations. It has been clearly shown that anterior and posterior parts of the digestive tract are formed through distinct processes from the posterior endoderm and the endodermal strand, respectively. The endodermal cells destined to the intestine move out of the tail much earlier before its absorption, which is also first described in the present study.
Summary of the Digestive Tract Morphogenesis in C. intestinalis
The digestive tract morphogenesis in C. intestinalis is summarized in Figure 7. The anterior digestive tract to form the esophagus and stomach develops through epithelialization, followed by folding and closure of the epithelia, in the two regions of dorsal and ventral in the central to right posterior endoderm facing the endodermal cavity; the dorsal region gives rise to the esophagus and the ventral region to the stomach. These morphogenetic events occur during stages 27 to 32 (Figs. 3, 4). By contrast, the intestine is formed in the left posterior trunk in a different manner. Cells to constitute the intestine originate from the endodermal strand in the tail; the cells in the endodermal strand move anteriorly into the left posterior trunk of a larva by stage 30, which occurs earlier than the absorption of the tail. During stages 30 to 32, the intestine is formed by accumulation and rearrangement of the cells originated in the endodermal strand (Figs. 5, 6). The duct of the pyloric gland is formed around stage 30 through bifurcation of the lumen left to the stomach (Fig. 3). Thus, the basic tubular structure of the digestive tract is formed by stage 32, followed by the differentiation of the distinct digestive organs that becomes evident during the body axis rotation stages. The digestive tract subdivisions are complete by stage 37.
Some Considerations on the Strategy to Describe Digestive Tract Morphogenesis
The strategy we took in the present study is a simple method combining confocal microscopy and computational rendering, focusing on the epithelial organization of the endoderm, a characteristic of the digestive tract. Feasibility of the method depends on the fact that epithelial organization of the digestive tract endoderm is easy to be identified by its morphology. At present, no antibodies that specifically stain endodermal epithelium in the larval trunk are available and genes specifically expressed in the endodermal epithelium of the developing digestive tract have been unknown in C. intestinalis. Considering these circumstances, the method applied can be assessed as reasonable.
Cells Labeled by pZip-Kaede Expression Construct
For tracing the endodermal strand cells, we used the pZip-Kaede expression construct. This allows labeling of the endodermal strand cells that express Kaede protein by local UV radiation at the tail.
In the larvae harboring the pZip-Kaede expression construct, approximately anterior two-thirds of the entire length of the endodermal strand emitted green fluorescence, when examined immediately after hatching. It has been reported that germ cell precursors derived from the B8.12 blastomere pair at the gastrula are localized in the distal region of the endodermal strand, expressing CiVH, the protein of the vasa gene homolog in C. intestinalis (Shirae-Kurabayashi et al., 2006). This region is likely corresponding to the region without Kaede fluorescence, and it has been shown that the germ cell precursors are carried into the trunk at the time of the tail absorption (Shirae-Kurabayashi et al., 2006). By contrast, the endodermal strand cells destined to the intestine move into the left posterior trunk much earlier than the onset of tail absorption, which is first described in the present study. Therefore, it is highly likely that the germ cell precursors are not included among the cells labeled by pZip-Kaede expression construct.
Anterior Two-thirds of the Endodermal Strand May Contain Some Distinct Cell Fates
Although the present study has clearly shown that the cells in the anterior two-thirds of the endodermal strand in the larva contribute to the intestine, red Kaede fluorescence was observed in other parts than the intestine. In some of the juveniles developed from the eggs introduced with the pZip-Kaede expression construct by injection or electroporation, red fluorescence was observed in a small vesicle structure, which was apparently distinct from the digestive tract tube and was located near the esophagus and the tail residue (Figs. 6G′–I′). Considering its morphology and location, the vesicle structure is presumed to be the periesophageal body, which has been reported by Okada and Yamamoto as the structure located just below the ventral margin of the esophageal opening, and filled with round vesicles and Golgi complexes, possibly with an endocrine function (Okada and Yamamoto, 1999). It is, however, unclear whether the cells constituting periesophageal body are of the endodermal strand origin. Additionally, it has also been reported by Shirae-Kurabayashi et al. that CiVH expressing cells derived from the B8.11 blastomere pair, the sister of the B8.12 blastomere, are localized in the anterior half of the endodermal strand, and that they coalesce into the intestinal wall but not the gonad after metamorphosis (Shirae-Kurabayashi et al., 2006). Nevertheless, the fate of those cells may be distinct from the neighboring endodermal cells as speculated by Takamura et al. (Takamura et al., 2002). Thus, the anterior two-thirds endodermal strand may contain some distinct cell fates, though a large part of the constituting cells are destined to develop into intestine epithelium.
Conclusive Remark
To understand how the digestive tract is formed is a key step toward understanding of the thorough process of the digestive tract development in C. intestinalis. Hence the present study will serve as an important basis for the future studies on morphogenetic as well as molecular mechanisms of the digestive tract formation, their diversity and evolution in ascidians and/or chordates.
Adult Ciona intestinalis were obtained at the Onagawa Field Research Center of Tohoku University and provided by the Maizuru Fisheries Research Station of Kyoto University and Misaki Marine Biological Station of University of Tokyo through the National Bio-Resource Project (NBRP) of the MEXT, Japan. Eggs and sperm were obtained surgically from the gonoducts. After insemination, fertilized eggs were washed and cultured in a Petri dish filled with artificial seawater or filtered seawater at room temperature (18–22°C). Developmental staging of larvae was done according to FABA2 (http://chordate.bpni.bio.keio.ac.jp/faba2/2.2/top.html). Juveniles were harvested after detaching from the Petri dish using a tungsten needle.
Phalloidin Staining
Phalloidin staining was carried out as described previously with some modifications (Christiaen et al., 2005). Larvae and juveniles were fixed with 3.7% formaldehyde in Ca2+/Mg2+ free seawater for 15 min. Fixed samples were permeabilized with the phosphate-buffered saline containing 0.4% Triton X-100 and 0.2% Tween 20 (PBTT1) for 10 min. To quench autofluorescence of aldehyde, samples were treated with phosphate-buffered saline containing 50 mM NH4Cl, 0.2% Triton X-100 and 0.1% Tween 20 (PBTT2) for 15 min. Then, samples were washed in PBTT2 for 5 min twice and were stained for 15 min with Alexa Flour 488 or 546 phalloidin (Invitrogen) diluted to 1:100 in PBTT2. Samples were further washed in PBST for 5 min twice and were immersed in VECTASHIELD with DAPI (4′,6-diamidine-2-phenylidole-dihydrochloride;Vector Laboratories). All manipulations were done at the room temperature. Stained samples were kept at 4°C in dark until microscopy.
Confocal Laser Scanning Microscopic Analysis
Stained larvae or juveniles were mounted in the center of a two-tiered sticky toroidal-shaped seal attached on the top of a slide glass. A cover slip was placed over the seal. Samples stained with phalloidin were visualized under a confocal laser scanning microscope (LSM5) with the LSM software, ZEN 2008 (Zeiss). An X20 objective lens was used. Image data were acquired in a z-stack mode (section thickness, 1.028 μm; pinhole, 75 μm; filters, LP 505; laser, 488 nm), (section thickness, 1.114 μm; pinhole, 81 μm; filters, BP 560–615; laser, 543 nm).
Confocal microscopic image data were analyzed using ImageJ software (provided by NIH). Three-dimensional constructions of epithelial tissues were done according to the procedure described on the DeltaViewer provided by Wada through HP: (http://delta.math.sci.osaka-u.ac.jp/DeltaViewer/index-j.html) (Wada, M., Osaka University, Osaka, Japan).
pZip-Kaede Expression Construct and Its Transgenic Line
Ci-Zip is a zinc transporter gene of C. intestinalis, expressing in the endodermal strand and a part of central nervous system (CNS) in tail-bud stage (aniseedV3_13279). To generate an expression construct that drives a given gene in the endodermal strand, an enhancer of Ci-Zip was amplified by polymerase chain reaction using primers (5′-TTTCTGCAGTGCTGACTCGGATGTATG-3′ and 5′-TTGGATCCTTGTGGAAAAGCATCAA-3′) as described by Shi et al. (2009). The endodermal strand enhancer was connected to the Ci-fkh basal promoter, which includes the upstream 148 bp from its initiation codon (Harafuji et al., 2002), and was inserted into pSP-Kaede (Hozumi et al., 2010) to generate the expression construct, pSPCiZipCifkhK (hereafter denoted as pZip-Kaede). pZip-Kaede drives expression of Kaede in the endodermal strand and a part of CNS from the tail-bud stage onward.
For generation of a transgenic line, a stretch of the nucleotide sequence including the endodermal strand enhancer, Ci-fkh basal promoter and Kaede coding region of pCiZipCifkhK was subcloned into pMiDestF (Sasakura et al., 2008) using a Gateway system kit (Invitrogen), and the plasmid, pMiCiZipCifkhK, was obtained. The transgenic line, Tg[MiCiZipCifkhK]1, was created by electroporation of the wild-type eggs with 60 μg of pMiCiZipCifkhK and 80 μg of Minos transposase mRNA. Selection of electroporated animals was performed as described by Matsuoka et al. (Matsuoka et al., 2005).
Electroporation of the Expression Construct
Electroporation of reporter constructs into fertilized eggs was carried out using a protocol adapted from a previous report (Corbo et al., 1997). For electroporation, a Gene Pulser Xcell electroporator (Bio-Rad) and electroporation cuvettes of 0.2 cm in width were used. Fertilized eggs were treated for 5 min in the dechorionation solution (sea water containing 1% of sodium thioglycolate, 0.05% actinase E, and 0.04N NaOH), thoroughly washed in fresh seawater twice and further washed in 0.693 M mannitol in 10% seawater twice. One hundred fifty microliters of egg suspension was mixed with 250 μl of DNA in mannitol (21:4 mixture of 0.77 M mannitol and 1 mg/ml plasmid DNA in TE buffer) to give a total volume of 400 μl and the plasmid DNA concentration of 100 μg/ml. The egg suspension was immediately transferred to a cuvette and exposed to an electric charge; 10 ms square pulse at 30 V. The eggs were returned to fresh seawater and reared at 18°C.
Exposure of a Larva Harboring pZip-Kaede to UV Radiation and Observation of the Larva and the Resulting Juvenile
A swimming larva developed from the egg introduced with pZip-Kaede construct was transferred to a drop of seawater mounted in the center of a four-tiered sticky toroidal seal attached on the top of a slide glass. To anesthetize the larva, it was resuspended in 20 μl of 0.2% MS-222 (tricaine mesylate) seawater. Then, the tail of the larva was irradiated with UV light using an Olympus BX60 microscope. The larva was returned to fresh seawater in a Petri dish and cultured at 18°C to the adhesion period or juvenile stage. For photographing, the juvenile was transferred into 7–10 μl of half-saturated L(-)-menthol in seawater mounted in the center of the four-tiered sticky toroidal seal attached on the top of a slide glass. Images were taken using the BX60 microscope equipped with an Olympus DP70 camera and processed by Photoshop (Version 7.0, Adobe Systems).
We thank the National Bio-Resource Project, MEXT, and members of the Education and Research Center of Marine Bioresources of Tohoku University, Maizuru Fishery Research Station of Kyoto University and Misaki Biological Marine Station, the University of Tokyo for providing us with adult Ciona intestinalis. Special thanks are due to the late Ms. K. Hirayama at Kyoto University for her devoted working for the NBRP. We thank Drs. K. Fukuda, N. Takatori, K. Yoshida, and T. Ikuta for discussions and Mr. H. Nezu for his work at the initial stage of this study. Thanks are also due to Ms. E. Zerinska for critical reading of the manuscript. TY was supported by Japan Science Society fellowship. H.S. was funded by Grants-in-Aid for Scientific Research (B) and (C) from JSPS (18370088, 22570207) and partly by JAMBIO.
- 2004. Development of Ciona intestinalis juveniles (through 2nd ascidian stage). Zoolog Sci 21:285–298. , , , , .
- 2005. A modular cis-regulatory system controls isoform-specific pitx expression in ascidian stomodaeum. Dev Biol 277:557–566. , , .
- 1997. Characterization of a notochord-specific enhancer from the Brachyury promoter region of the ascidian, Ciona intestinalis. Development 124:589–602. , , .
- 2006. Tunicates and not cephalochordates are the closest living relatives of vertebrates. Nature 439:965–968. , , , .
- 2002. Genome-wide identification of tissue-specific enhancers in the Ciona tadpole. Proc Natl Acad Sci U S A 99:6802–6805. , , .
- 2000. Developmental fates of larval tissues after metamorphosis in the ascidian, Halocynthia roretzi. II. Origin of endodermal tissues of the juvenile. Dev Genes Evol 210:55–63. , .
- 2010. Efficient transposition of a single Minos transposon copy in the genome of the ascidian Ciona intestinalis with a transgenic line expressing transposase in eggs. Dev Dyn 239:1076–1088. , , , , , , , .
- 2004. Ciona intestinalis Hox gene cluster: its dispersed structure and residual colinear expression in development. Proc Natl Acad Sci U S A 101:15118–15123. , , , .
- 2005. Germline transgenesis of the ascidian Ciona intestinalis by electroporation. Genesis 41:67–72. , , , , .
- 1999. Differentiation of the gonad rudiment into ovary and testis in the solitary ascidian, Ciona intestinalis. Dev Growth Differ 41:759–768. , .
- 2008. The amphioxus genome and the evolution of the chordate karyotype. Nature 453:1064–1071. , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .
- 2008. Enhancer detection in the ascidian Ciona intestinalis with transposase-expressing lines of Minos. Dev Dyn 237:39–50. , , , , .
- 1994. Developmental biology of ascidians. New York: Cambridge University Press. .
- 2003. The ascidian tadpole larva: comparative molecular development and genomics. Nat Rev Genet 4:285–295. .
- 2009. FGF3 in the floor plate directs notochord convergent extension in the Ciona tadpole. Development 136:23–28. , , , .
- 2006. Dynamic redistribution of vasa homolog and exclusion of somatic cell determinants during germ cell specification in Ciona intestinalis. Development 133:2683–2693. , , , , , .
- 2002. Primordial germ cells originate from the endodermal strand cells in the ascidian Ciona intestinalis. Dev Genes Evol 212:11–18. , , .
- 1995. Coordinated expression of Abd-B subfamily genes of the HoxA cluster in the developing digestive tract of chick embryo. Dev Biol 169:76–89. , , .
- 2008. Left-right asymmetric expression of Pitx is regulated by the asymmetric Nodal signaling through an intronic enhancer in Ciona intestinalis. Dev Genes Evol 218:353–360. , .
- 2011. Repression of Rx gene on the left side of the sensory vesicle by Nodal signaling is crucial for right-sided formation of the ocellus photoreceptor in the development of Ciona intestinalis. Dev Biol 354:144–150. , .
- 2012. Establishment of enhancer detection lines expressing GFP in the gut of the ascidian Ciona intestinalis. Zoolog Sci 29:11–20. , .
- 2009. Vertebrate endoderm development and organ formation. Annu Rev Cell Dev Biol 25:221–251. , .


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This topic has been moved to The Yorks.
The British Royal Family *Windsor* / Kate Middleton / MOVED: DM: the York Princesses are ugly inbreds compared to Waity.
on: Today at 08:56:01 pm
|Started by Alexandrine - Last post by Alexandrine
This topic has been moved to The Yorks.
The British Royal Family *Windsor* / The Yorks / Re: DM: the York Princesses are ugly inbreds compared to Waity.
on: Today at 08:45:21 pm
|Started by Stephanie - Last post by Val
QE's head in the sand attitude in not dealing with the situation will contribute to the inevitable and predicted end to the Monarchy.
on: Today at 08:02:43 pm
|Started by dianab - Last post by HRH Tiana of NOLA
on: Today at 07:38:37 pm
|Started by Mia - Last post by Countess of Holland
Agreed and I think that that person had more support than Letizia who had to deal with her father-in-law's disapproval. And I know from my own experience that that can be very daunting. The mother of my former boyfriend was quite clear that she felt I wasn't good enough for her son, she had wanted her son to hook up with the daughter of one of her tennis-friends.
And that disapproval ate me up, and that is just at a regular family who is not covered by the press and I didn't have to work with her, she wasn't my 'boss'.
I think that it is one of the reasons why I have a soft spot for Letizia. I admire her for how she has handled it all and how she is holding herself, both as Princess of Asturias and now as Queen.
BTW When it comes to body language, the world of the Borbons and the Cambridges couldn't be further apart. Whenever King Felipe looks at his wife and talks to her, he has a smile (sometimes big, sometimes a faint smile) on his face. Whenever William looks at Kate, he has clenched jaws.
The British Royal Family *Windsor* / The Yorks / Re: DM: the York Princesses are ugly inbreds compared to Waity.
on: Today at 07:31:02 pm
|Started by Stephanie - Last post by Countess of Holland
^^I think we can forget the Queen taking action. She is notorious for not taking action or at least wait for it for ages. The RF is petrified that the days of the War of the Wales' and the days after Diana's death will return and they will bend over backwards to the Middletons to make sure it won't happen.
If something is to be done about the odious Middletons, it will be under King Charles III (or whatever regnal name he will choose) but not sooner.
on: Today at 06:32:17 pm
|Started by Mia - Last post by gingerboy24
I think she has done a grand job, and especially since becoming Queen Letizia. She mades a few mistakes with her clothes, but then who doesn´t. Overall she has done incredibly well and, for me, comes across as regal without being uppity about it. I admire her, she does her best, she tries, and that is more than some else I could mention...
The British Royal Family *Windsor* / The Yorks / Re: DM: the York Princesses are ugly inbreds compared to Waity.
on: Today at 06:28:31 pm
|Started by Stephanie - Last post by gingerboy24
Oops, meant to say in my post two above, end of first line, council cath not glamorous - having said that council caro is not either.
The British Royal Family *Windsor* / The Yorks / Re: DM: the York Princesses are ugly inbreds compared to Waity.
on: Today at 06:12:25 pm
|Started by Stephanie - Last post by Kuei Fei
I'm not sure what fiendish delights the courtiers fantasize about, but I HOPE HM has the guts to punish Kate one way or another. There is ZERO excuse for something like this and frankly, if I were around Kate regularly, I would be on edge and tense.
Kate strikes me as one of those unstable, testy types that'll go haywire on a person over the smallest thing.
Their problem is that nowadays we all see through, know which direction the nastiness is coming from, and why. In effect, all she is doing is digging herself a deeper hole all the time. Coucil caro is not glamorous, not by a long shot, and that cosmetic surgery she keeps having is turning her face into something awful, changed beyond recognition, and not for the better. How would she like it if the tables were turned. Not one of us have had anything to do with how we look, our features are there when we are born. If council caro got this article on the go then people in glass houses should not throw stones. Pa medd not exactly good looking, council caro looks as mean and nasty as they come, cupcake looks away with the fairs and is a very unattractive shemale, orangine looks as though she had a fight with the back of a bus and the bus won. Fester not exactly good looking either is he, another ugly bug. To call the Yorks out like this, in public and with such venom is unnecessary and just so nasty and vile it is untrue - I read it and it made my blood boil. They took bullying from her, put up with her nastiness, and now she is turning the tables. Council cath has never made any effort with the rf, not even before the engaement, and even less now, and getting worse all the time. Thumbing her nose up and traditions going back centuries, disrespectufl at the Cenotaph, refuses to learn any royal etiquette whatsoever, flashing her nake lady bits to the world. Just who do coulcil cath and caro think they are, that they can try and belittle the Yorks, or anyone else for that matter. They are not the ones bringing the monarchy down the ugly bug meddledoom family are..
I do hope HM hauls her and coouncil caro over the coals for this, they have to be behind it, nobody else would want to be that vicious towards them. As for the old bruiser who wrote the article, she should also be ashamed of herself, she needs to look at herself long and hard in front of a mirror, she most certainly lives in a glass house and deserves to have stones thrown at it. Hope she does not rely on her journalism for an income, she has done herself a great disservice with this article, and I for one have minus zero sympathy for her.
I swear that woman is nothing but trouble and can't seem to stop making trouble.
First, the Yorkies are blood princesses and while that might not seem to be much in terms of succession, but they were there first in William's life and I don't believe that somehow they're somehow less worthy than Kate and her dippy husband. Second, they aren't causing half the messes that Kate and William has and since WK are higher up, the messes that Kate and William make have a more damaging effect on the monarchy. Third, Kate has to be the only member of the BRF since Fergie who has made so many enemies and frankly can't seem to stop causing trouble all the time. One thing after another comes from that woman and still, Kate and her DM toadies bash the Yorkies and anyone else who doesn't lick Kate's boots.
Out of the entire RF, the Yorkies have caused HM the LEAST trouble and have really gotten it together.
The British Royal Family *Windsor* / The Yorks / Re: DM: the York Princesses are ugly inbreds compared to Waity.
on: Today at 06:03:00 pm
|Started by Stephanie - Last post by Little light
I think that article was, to steal a phrase from someone else, "the longest suicide note ever written".
And I think they, the Medds, will regret it.
We can choose our friends but not our family, so it would have been in the Medds' interest to cultivate friendship within the BRF as it would, or might, have paid dividends in the long term. But as always with that family, it is always about themselves. NOBODY else matters.
I hope they are dealt with severely.,
Royal Families / Swedish Royal Family / Re: Crown Princess Victoria expecting second baby in March 2016
on: Today at 05:56:46 pm
|Started by beline - Last post by HRH Tiana of NOLA
Oscar really favors Queen Silvia.


text:
On this Date in…
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- 2012: Raven Show #4: Bird Language
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Loon Identification Update
I have had a few responses to the Loon Identification post I made a couple of days ago. One of them pointed out that there is a discussion of Loons in the Peterson Field Guide to Advanced Birding by Kenn … Continue reading
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Hotfrog INDIA provides information regarding ANALYTICAL AUTOMATION (I) PVT.LTD. in Andheri(East) MAHARASHTRA. ANALYTICAL AUTOMATION (I) PVT.LTD. is located at 504 Twin Complex Fire Brigade Road Marol, Andheri(East) MAHARASHTRA 400059 and provides Haematology services. Contact them on 28202988.
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When ever we talk about promoting blog on various network, one of the wisest advice you would get is "Promote Website on Facebook". A Social-networking site with 900+ million active users, it's one of the easiest way to promote your website viral for free or for minimum cost. Blogging isn't just about blogging any more. It's about making lasting, sincere connections with people. While the blog is still the main platform, it's not as easy as it once was to build an audience on a blog alone. People are discovering sites in many ways now, including social (13 comments)
10 Ways to Drive Traffic From Forums To Your Blog?
I'm only spending about 10 minutes a day on discussion forums, but from this little effort, I have successfully managed to drive traffic from forums every single day to my blog. It's all because I have a plan, and I stick to it. It's about adding quality content, but it's also about 10 different things. <Image Credit> I'm active In order to get traffic from discussion forums you should be present and publish something at least a few times a week. The more active you are, the more traffic you'll get. I publish something 3 times a day, every (41 comments)
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Viral content is best way to grab attention and attract traffic to your blog. An article went viral means, that article got great number of share and traffic. This could be on any traffic source for example, Stumble Upon, Digg, Facebook, Twitter, Google and so on. Creating viral content and making it reach to mass is not an easy task but if you manage to do so, you can't always expect a viral content to get lots of hits and popularity. Viral content are something which people always talk about for example a list posts like 101 ways to do something and so (20 comments)
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Excellent service and products
This is the 4th time I have used Garden Grocer. They have saved me tons of money while I have stayed on Disney Property. They always show up when they say and even when I screwed up my reservation dates they went out of their way to see I was taken care of! Great Job guys!!!! Takes so much!! Steve M. Chicago Ill
It was so easy to order our groceries! Everything was delivered on time, and without any problems. I also liked that we got a text message when our groceries were delivered. Will definitely use Garden Grocer again!


text:
Counterfeit money hunt lead to Cypress
HUNTINGTON BEACH – Police arrested a Corona man and a Garden Grove woman Thursday during an investigation into a counterfeit currency operation that led Huntington Beach detectives and United States Secret Service agents to a motel in Cypress.
Members of a task force serving a federal search warrant arrested Salvador Serrano Jr., 40, on suspicion of a variety of counterfeiting charges and Christine Ann Harrison, 43, on suspicion of drug possession, according to a Huntington Beach police statement.
The task force members found "a number of counterfeit bills in the process of being manufactured, along with all the equipment needed to complete the process," police said.
Authorities did not specify how much counterfeit money was found, how they initially identified Serrano as a counterfeiting suspect or what led them to the motel room.
Serrano was taken into federal custody, authorities said, while Harrison was booked into the Huntington Beach city jail.
The Huntington Beach investigators, along with numerous other local law enforcement agencies, are members of the U.S. Secret Service's Los Angeles Electronic Crimes Task Force, which focuses on high-tech crimes.
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Winner of CBD deal in at start
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The firm awarded the contract to design the upgrade of the Invercargill central business district had inside knowledge of what was required, having earlier been subcontracted to help produce a broadly based CBD "outline action plan".
The revelation raises questions around the integrity of the process, but those involved insist it was above board and fair to all parties.
The Invercargill City Council announced last month that Christchurch company Pocock Design:Environment had been chosen to design the upgrade of the Invercargill CBD.
The council had set aside $130,000 for the work.
Pocock Design had been in a three-way battle to get the contract with Queenstown's Baxter Design Group and CCM Architects Ltd, of Wellington, also in the running.
Pocock Design boss Craig Pocock, who said he was excited to get the contract, confirmed yesterday his company had also been involved in putting together the original CBD outline action plan document for public discussion. His firm had been subcontracted by Urbanismplus to do the landscape design portion of the work, he said.
Now his company has been awarded the contract to design the CBD upgrade, the roles will be reversed.
Mr Pocock said yesterday his company would be using Urbanismplus to make sure they stayed on the right track when designing the work.
His company had openly competed against some of the biggest firms in New Zealand to win the contract, he said.
"We just had the better proposal . . . I don't think we got it because we worked with them [CBD Working Group] previously. We never assumed we had the inside running at all."
Kobus Mentz, of Urbanismplus, did not return calls yesterday, but a staff member said its future involvement in the CBD upgrade would be quite small.
Invercargill City Council director of works and services Cameron McIntosh said all the bidders for the design work were given the same documentation to work from before pitching their design plans.
"The inner-city working group chose the successful one on its merit and there has been no information withheld and the successful person [Mr Pocock] was very open about his involvement both before and after," Mr McIntosh said.
CBD Working Group boss Norman Elder said yesterday Pocock Design had the advantage of being involved in the initial process for the CBD work, but said all contenders were judged on their merits when the contract was awarded.
He preferred Pocock Design's bid because it had a good understanding of how the inner-city working group wanted to make sure the CBD design worked for businesses in the area, he said.
Baxter Design Group boss Paddy Baxter, whose company lost out on the contract, said they were given a fair crack at winning the contract.
CCM Architects director John Rogers said he also had no problems so long as any conflicts of interest were declared and made obvious to the people making the decision.
Meanwhile, Mr Pocock is due in Invercargill today to discuss the way forward for the CBD upgrade design work, with community consultation to begin early next year.
- The Southland Times
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List all words ending with umpire
Only the word umpire was found, nothing longer.
Some random words: uvarovite chablis lea eke diureses voes llama
This is not a dictionary, it's a word game wordfinder.
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Associated Press
GLEN CANYON NATIONAL RECREATION AREA — A proposed off-road vehicle management plan for Glen Canyon National Recreation Area and the future of Lake Powell invasive mussel prevention are subjects of five open houses in February.
National Park Service staff will provide information and gather public comments on these and other current planning topics.
Each open house is from 4 to 7 p.m. in: Blanding, Tuesday, Blanding Arts and Events Center, 639 W. 100 South; Escalante, Wednesday, Escalante Interagency Visitor Center, 755 W. Main; Kanab, Thursday, Kanab Middle School, 690 S. Cowboy Way; Page, Ariz., Tuesday, Feb. 11, Glen Canyon National Recreation Area Headquarters, 691 Scenic View Drive; and Salt Lake City, Thursday, Feb. 13, Utah Department of Natural Resources, 1594 W. North Temple.
An electronic copy of the Glen Canyon plan may be downloaded at parkplanning.nps.gov/glcaorvplan.
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iTunes Ping goes live on iOS 4, not yet iPad
In addition to the release of iTunes 10 for Mac and PC earlier this evening, Apple has brought the Ping social music networking feature to the free iTunes application for iPhone and iPod touch. Rather than offering Ping as a standalone application, the company has integrated most of its features into a button that's now found at the bottom of iTunes, using button-style tabs to let you switch between a timeline of Activity, lists of People you follow and who follow you, plus Profile, which contains a seemingly non-editable version of the profile you create with the desktop application.
Notably, the Ping button does not appear as of yet in the iPad version of iTunes, and may not do so until the November release of iOS 4.2.
iLounge's Editor-in-Chief Jeremy Horwitz welcomes iLounge readers to follow him on Ping as he tests the service.
Update: Ping does not appear to work on iOS 3 devices, including original iPhone and iPod touch models, unupdated iPhone 3G/3GS or iPod touch models, or the iPad.
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Tell me the button bar is still configurable and that I can remove this Ping nonsense from it. Why do I need (yet another) "social network," and why does it have to be in my music?
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 2, 2010 at 11:02 AM (CDT)
When does the ping appear on the bottom bar? I was hooked to itunes last night, and got no update to the iphone 4, and I don't have the ping button today.
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 2, 2010 at 11:22 AM (CDT)
So this might be a stupid question but I'm genuinely curious—how did Ping appear in the iTunes app without my having to update it or install iOS 4.1? It just appeared by itself, automatically… but how? And does this mean that any app has this ability?
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 2, 2010 at 12:55 PM (CDT)
@ttman21 , @Farnsworth It appears in the iTunes Store app, not iPod.
@gouged the iTunes store app just pulls in web-based content from Apple. It doesn't need a software update to change.
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 2, 2010 at 3:10 PM (CDT)
I've not downloaded iTunes 10 yet. I used to use the LaLa.com service quite a bit as it was an awesome way for me to discover new music, especially when my chosen genre of music is just not available in the typical mainstream or "Top-40". The closest would be Disturbed or Iron Maiden. After all, what is the likely I'll see Kamelot or Sonata Arctica on mainstream FM radio? Anway, even though I pretty much ignored the whole social network thing (despite much badgering from friends, I still refuse to sign up on Facebook, for example), but I did like LaLa quite a bit, even to the point that I had some 10,000 tracks, nearly all power and progressive metal (sorry, but I just simply cannot stand rap or hip-hop, nor do I do country), uploaded to the service for streaming. I found all kinds of awesome new music on there as a result as recommendations by "followers" and those that I "followed". Therefore, I am curious as to what Apple ultimately did with Lala, as I am quite certain that this whole Ping thing was a result of Apple buying out LaLa.com.
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 2, 2010 at 11:13 PM (CDT)
Finally, I find an article explaining that the iPad does not yet have Ping. I searched expecting it to be there of course. How weird that it would not be on the iPad of all things. Even weirder that Apple does not explain anywhere that iPad owners have to wait. Anyhow, thanks for finally answering my question….
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 6, 2010 at 10:14 AM (CDT)
Apple is relying on iOS 4, and even iOS 4.1 in some cases, to implement these new features. The iPad is of course still running iOS 3.2, so it's being left out in the cold for now.
TV Show Rentals are another new feature that isn't yet supported on the iPad, either.
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 6, 2010 at 11:46 AM (CDT)
I can understand that insofar as getting the content/data but how does it know to appear as a button in the button bar with regular iOS SDK UI elements? Aren't these in the Objective-C iOS SDK and would need to be added in the app as code and compiled, etc to show up?
For example, this seems to imply that it's possible to just write an app without any interface at all and then just pull in stuff from the web that creates the entire interface (buttons, textboxes, everything) including functionality? This is what this Ping addition to iTunes app seems like it did so it sounds like you can bypass Apple approval process using this method then?
Posted by Jeremy Horwitz in East Amherst, NY, USA on September 8, 2010 at 6:38 PM (CDT)
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Indiana State University Professor Matthew Brennan has completed work on his fifth book of poetry.
It was published in late July by Backwaters Press.
Brennan began work a decade ago on his 84-page collection titled "The House with the Mansard Roof." It includes poems that have appeared individually in other publications throughout the last several years.
Some of this collection of Brennan's work was influenced by artwork created in the 1930s and 40s by American artists commissioned by the U.S. Works Progress Administration.
Prints of those Works Progress Administration paintings hang on the walls outside Brennan's office in an upstairs hallway of Root Hall on the ISU campus.
But because Brennan is a poet whose creativity is spurred by his own personal experience, much of this work has a more personal connection for him. For example, the poem after which the book was named was inspired by a neighbor's house that Brennan views from one of his own upstairs windows.
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This new work offers a glimpse of Brennan's more vulnerable side, he says. And because he discarded a number of previously published poems to assemble the best of his personal best, he likes to think "The House with the Mansard Roof" is a book that "holds up."
It is a collection that captures the essence of why Brennan writes.
"When I'm writing a poem that works, I feel more fully myself then than at any other time," he said.
Brennan, who completed his undergraduate work at Grinnell College, earned his master's and doctoral degrees at the University of Minnesota. He is a distinguished honorary member of the National Society of Collegiate Scholars whose works have appeared in more than 60 publications.
"The House with the Mansard Roof," published by The Backwaters Press, is available through the publisher or at barnesandnoble.com and Amazon.com.
Writer: Rachel Wedding McClelland, Indiana State University, assistant director of media relations, at 812-237-3790 or email@example.com
Backwaters Press publishing has released a collection of Indiana State University English Professor Matthew Brennan's poetry titled "The House with the Mansard Roof."
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Judy Heise's infant daughter died from SIDS in 1969, and for years Judy searched for answers as she struggled with her fractured faith. This is Judy's journey to healing and reconciliation with God.
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Alice Through the Looking Glass
There is no magic, no wonder, just junk rehashed from a movie that was itself a rehash of Lewis Carroll, tricked out with physically unpersuasive…
There is no magic, no wonder, just junk rehashed from a movie that was itself a rehash of Lewis Carroll, tricked out with physically unpersuasive…
Bécue-Renard brings his own brutality to the topic of PTSD, by putting us at odds with feeling his subjects' pain, or only studying it.
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Patrice Leconte's "Monsieur Hire" is a tragedy about loneliness and erotomania, told about two solitary people who have nothing else in common. It involves a…
Ken Loach's "I, Daniel Blake" wins the Palme d'Or at the 69th Cannes Film Festival.
Barbara Scharres on the cats of Cannes 2016.
Roger's Favorites: Werner Herzog.
An article about the 2016 ILLUMINATE Film Festival in Sedona, Arizona.
Part two of Jana Monji's essay about the portrayal of Asian characters in cinema.
Part one of Jana Monji's essay about the portrayal of Asian characters in cinema.
A preview of the 2016 Chicago Underground Film Festival.
A review of the History Channel remake of the landmark mini-series, "Roots."
* This filmography is not intended to be a comprehensive list of this artist's work. Instead it reflects the films this person has been involved with that have been reviewed on this site.
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Mike Nichols' 1971 drama "Carnal Knowledge" is part of a canon of American films of the late 1960s to mid-1970s that mirrored the freewheeling sexual culture and society from which they emerged. These films ("Bob and Carol and Ted and Alice", "Shampoo" and others) examined varied notions of commitment, companionship and sex. "Carnal Knowledge", written by playwright, author and cartoonist Jules Feiffer, shows men talking casually, bluntly and frankly about women, their bodies, of strategies to get sex and of sexual belt-notching, though not necessarily much about the specific act of sex.
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Q. I was watching "Godfather II" the other day and remembered that the scene with Vito sitting on the steps with his family, after he has killed Fannuci, was originally intended to lead into an intermission, which was scrapped somewhere along the way. I'm pretty sure James Cameron never intended on having an intermission for "Titanic," but let's pretend the studio asked you, an up and coming editor, to find a place to insert one. Even though you know it was wrong and that Cameron would surely kill you, where would you have put an intermission? (Frank Mendez, Dallas, TX.)
Louis Malle, who died last week at 63, was a director whose movies caused scandal sometimes for their content, sometimes for their style, sometimes for both. The respected French filmmaker, married since 1980 to actress Candice Bergen, died Thursday at their home in Los Angeles, from lymphoma.
If this were a scene in a movie, you'd know right away that it was the scene about the actress' last day in town. Jacqueline Bisset's two-room suite at the Sheraton Plaza was scattered with props for her departure. The bedroom floor was lined with trunks and suitcases, their lids tilted open, clothes tumbling out of them. Leaning against the wall in the living room was one of those big cartoonist's caricatures showing Jackie with a giant head balanced on top of a tiny body. It said, "Good Luck, Jackie!" and it was signed by all the members of the cast and crew of her latest film.
"Kramer vs. Kramer," the drama of a child custody fight, won nine Academy Award nominations on Monday – and so, in a surprise, did "All That Jazz," about a Broadway director's self-destruction.
In the beginning it didn't much matter where you made a movie. The motion picture was a gimmick nobody took very seriously, and the vaudeville houses used them to chase out customers between shows. When the customers started to linger, an industry was born.
CARTAGENA, COLOMBIA - "There's no use kidding ourselves," Lewis Gilbert said. "The appeal of this film will be based on sex and violence, of course. But the difference is, this will be the first sex and violence epic. Usually sex and violence are counted on to sell themselves, and so that sort of thing is made cheaply..."


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Arthur J. Gallagher & Co. (NYSE: AJG) announced the acquisition of G. S. Chapman & Associates Insurance Brokers, Inc. (Chapman Insurance) headquartered in Pasadena, California. Terms of the transaction were not disclosed.
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