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Wow. So I have been working very hard to drop all of this extra weight I've been carrying around the last couple of years and am very proud to say, even broadcast that I went from 172 to 154. I have a goal to get to around 145 and stay in that vicinity - so still about 10 pounds to go - but I can't tell you how much better I feel about myself and how big a difference it makes in feeling confident.
Here's how I did it (so far):
1) Eat WAY less. Typically I have 2 slices of toast (diet bread at 40 calories each) with 1 tbsp (that's 1/2 a serving) peanut butter with coffee for breakfast. For lunch, I usually have a cup of soup (I measure it - a can has 2 or more servings - don't eat the whole can) with rice crackers or rye crisps, a wedge of Lite Laughing Cow cheese and an apple. I also only eat low sodium versions of soup - writing down what I eat everyday and weighing everyday I have learned EXACTLY what makes me fat. For me it's salt and sugar. Dinner depends on my schedule as I may only have time for a Lean Cuisine (nothing over 325 calories). If I don't have a show, I'll eat very specific portions of fish or chicken (3 oz) with a full cup of steamed vegetables and something like brown rice or couscous. I also cut way back on alcohol - another food group that really sends the scale tipping.
2) I'm active EVERY. DAY. I either work out on the elliptical machine, do a pilates routine (sometimes both) or I am in a very physical sketch comedy show that includes dancing, running around and a scene where I have to hold my arms up the sides (straight out) for about 5 minutes (this has been a huge help!)
3) I am dilligent in making healthy choices at restaurants. Less a slip or two here or there I will order grilled vs. fried, fruit vs. fries, salad vs. sandwich, etc.
4) I didn't get carried away during Thanksgiving and worked out on Thanksgiving. No seconds (of anything).
5) I eat an apple a day. It's full of fiber and requires a lot of chewing which helps me to feel full and satisfied.
6) I eat my meals sitting down, and chew slowly.
7) I do not eat while I cook. I chew gum.
8) If I'm hungry I will drink a full glass of water - I believe I have minsinterpreted thirst for hunger my whole life.
9) I try not to ever eat late. (I've slipped here a couple of times because I couldn't eat before shows that I'm in, but for the most part I'm done eating by 6:00.)
10) I have publicized and shared my diet plans and progress with supportive friends and now here too. This accountability helps me to stay focused and the possible embarrassment of gaining the weight back would be that much worse.
Here's to celebrating the holidays not worrying about what I'll wear. I have a new red dress that is 2 sizes smaller than last years hanging within view of my exercise machine. It reminds me of what I've been working for and don't want to see slip away again. I had to ask the saleslady twice to bring a different size - what a happy day.
Tuesday, December 8, 2009
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International Rijeka Carnival on Korzo with many participants and onlookers. Source: index.hr
Trsat Fortress, or a medieval fortress with the Marian sanctuary nearby. Source: flickr.com
Rijeka - port, merchant and tourist city with numerous hotels and apartments in downtown Rijeka and its surroundings. Source: croatia.hr
Key tags: private and hotel accommodation in Rijeka, vacation in Rijeka, destination description, events, sights, rooms and apartments Rijeka - Croatia.
Other destinations of Primorje-Gorski Kotar County: Banjol/Rab | Barbat/Rab | Baska/Krk | Crikvenica | Dramalj | Fuzine | Icici | Island of Cres | Island of Krk | Island of Losinj | Island of Rab | Jadranovo | Kampor/Rab | Klenovica | Kostrena | Lopar/Rab | Lovran | Malinska/Krk | Matulji | Medveja | Moscenicka Draga | Njivice/Krk | Novi Vinodolski | Opatija | Punat/Krk | Risnjak NP | Selce | Silo/Krk | Supetarska Draga/Rab
Room Sarah, Rijeka
Kalvarija ulica 14
Apartment Dvornicic, Rijeka
Andrije Medulica 1
Apartment Ajbek, Rijeka
Pionirska 23
Apartments Aster, Rijeka
Drazicka ulica 23
Apartment Veronika, Rijeka
Istarska 60
Apartments MB, Rijeka
Ulica Ivana Lenca 14
Apartment Brajin, Rijeka
Janeza Trdine 1
Apartment Angela, Rijeka
Istarska 30, Kantrida
Apartment Corallium, Drenova
Bok ulica 16B
Apartments Marija, Rijeka
Jurja Dobrile 19
Apartment Ada, Rijeka
Pionirska ulica 23
Apartment Rosa, Rijeka
Mirka Jengica 6
Apartments Costabella, Rijeka
Opatijska 33
Apartment Tijara, Rijeka
Ulica Ede Starca 18
Apartment Filodrammatica, Rijeka
Korzo 30
Apartments Skoljka, Rijeka
Creska ulica 9
Apartment My City, Rijeka
Matije Gupca 12

subset: commoncrawl


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Friday night I went and purchased some sidewalk chalk, claiming that it was an activity for Will, but in all actuality I just really like to play with sidewalk chalk. It was fun, except for the part when Brett & I had to lay down and be "traced"... I looked a little like my elementary school teachers after that, with chalk down my outfit.
On Saturday morning we got in the Jeep and headed to Yakima (or near Yakima) to visit Brett's Grandparents. Will had been wanting to see the animals that they had living nearby. The drive was about 3 hours. I think we were about 15 minutes in when Will said "Um, guys?... This is taking a reeeeallly long time..." :)
This was a cool gas station on our way, shaped like a teapot. We knew for sure it was not in working order when we saw that gas was $1.79/gallon :)
Brett's Grandfather took us to a ranch nearby their home where we saw horses, goats, sheep & cows, & some big tractors & trucks.
This was out back behind B's Grandparents home. There were a few horses & a little Colt.
(I could have used some sunglasses)
On our way home, we stopped at a yummy burger place in North Bend.
The jeep has bugs all over the windshield so when we were approaching home, Will woke up suddenly and said "The Jeep! we need to wash the Jeep!" So we did. :)
On Sunday Will went to his momma's house. After church, B & I went on a bike ride along the Slough. It was such a gorgeous day for it too. I am so incredibly out of shape though... Something that becomes evident when riding my bike.
After our ride, we headed to my Aunts house for dinner & time with family & new friends... Which in my family apparently means rockband & a game in which one sprays whipped cream into their palm, slams their wrist & tries to catch the whipped cream in their mouth. I was not very good at the game and ended up with whipped cream all over my skirt.
On Monday morning we got up early to go play Rugby in Seattle (I, of course, was a sideline cheerleader, as I play no sports and lack the coordination necessary to do so).
After Rugby, we went to Mike & Jen's apartment in the sky (which is a-mazing) & they made us a delightful breakfast :)
We then went to check out the Northwest Folklife Festival that was going on. There were a ton of people there... And oh what a people they were! People-watching at it's best :)
After the days festivities, we went to M & J's for a BBQ, then headed to my apartment, where we went swimming/hot-tubbing a bit.


text: Stubborn NM Fire Burns 50 More Homes After jumping lines, blaze is 35% contained By Harry Kimball, Newser Staff Posted May 3, 2008 6:52 PM CDT 0 comments Comments A fire near Roswell, New Mexico, March 14, 2008. (AP Photo) View 2 more images (Newser) – Firefighters nearly contained a New Mexico wildfire this week before it leaped containment lines and razed another 50 homes on Wednesday, the AP reports. Nine had already gone up in flames. Firefighters are working today to cut new lines around the blaze, which is 35% contained near Albuquerque. Officials say the 13,790-acre fire was man-made. View 2 more images


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Thursday, 18 September 2014
The best crew
I tend to keep my communications personal, preferring to talk in person. Somehow, it feels more heartfelt to me, and besides... I'm a private sort of person.
The reflective blogs Ive written these last few days have been the result of an outpouring inspired by the incredible shooting experience of Learning to Breathe.
This last one, for a while, finishes a series of blogs with something very dear to my heart. The crew.
I've worked with all sizes of crew over the years, with varying degrees of experience. but, hand on heart, I have never felt so connected to a crew as I have done with this film.
Many of them are people I have known for sometime.
I began putting together my core crew who came to Tobago with me back in February.
These were people who I trusted, loved and wanted around me during a shoot I envisaged as being intense both physically and mentally.
I didn't ask around. I wanted these people, and I was beyond lucky that they came straight onboard and understood exactly what I was trying to do.
In some cases the crew were stepping way outside their comfort zones, in a professional sense, but not once did they falter.
When we arrived in Tobago, the family atmosphere, I had hoped to foster, was immediately evident, and a strong unit was quickly formed that seemed utterly unbreakable.
Being in a somewhat undeveloped country it wasn't all plain sailing. loss of electricity and water plagued our days, but not once did I hear a complaint or a lack of enthusiasm for the job. Extraordinary.
My own relationship with each of these wonderful people is already known to them, so I won't go into this private area, but suffice to say I owe each of these people a debt i can never hope to repay. Although I will give it a bloody good go!
A month later we were back in London, and the core crew were joined by a larger group of people, adding to the skills and expertise onset.
Every single person on that set was either selected by me or recommended by a trusted source.
Every single on of those crew rocked very very hard.
In to the mix were a contingent of students who were seeking experience on-set.
I can say with certainty that ALL of these students will be bright shining stars of the future.
their professionalism and enthusiasm was extraordinary, and I will be keeping tabs on each and every one of them!
So, this reflection on working with an amazing crew, ends this current set of blogs, as I head into post-production on the film.
My next film looms large, and with a substantially bigger budget and bigger canvas to play in, I would have no hesitation in recommending every single one of my Learning to Breathe crew to the producer.
If you are a production manager or line producer and you are looking for brilliant crew members get in touch. I have some gems for you!
at 01:44
Labels: crew, crewing, dan turner, drama, feature film, film, films, learning to breathe, london film academy, NUA, shoreline entertainment, unit
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Little Mini Cakes are great for Bridal showers and bridesmaid gifts with a tag that says will you be my bridesmaid. Check out our website at www.cakesbyshara.net for contact information.
Saturday, January 31, 2009
Monday, January 26, 2009
Blog Luv Award
Blog Luv Award
Tara with Lavish Events sent me a blog Luv award and I want to pay it forward.
Thank you Tara!!
Here are some of the blogs I love.
Gray Photography Blog
Rhonda Patton Designs
Duce2Duce Wedding Videos
A Magical Affair
Ashely's Bride Guide
A Delightful Day
Lavish Events
Get Relish
Daisy's Favorite Things the Fabuless Newleywed
Brocade designs
Here are some of the blogs I look at daily. Congrats to you guys, and keep paying it forward!
Honorees please follow these instructions and share the LUV!
1. Put the logo on your blog or post.
2. Nominate at least 10 blogs which show GREAT ATTITUDE and/or GRATITUDE
2. Nominate at least 10 blogs which show GREAT ATTITUDE and/or GRATITUDE
3. Be sure to link to your nominees within your post.
4. Let them know that they have received this award by commenting on their blog.
5. Share the love and link to this post and to the person from whom you received the award.
Posted by Cakes By Shara at 3:00 PM No comments:
Saturday, January 17, 2009
Monday, January 12, 2009
Let's Eat Cake
Masterpiece Weddings, an event coordination agency founded by Melissa Margarita-DiStefano, is working really hard to put on an amazing fundraiser in February called Let's Eat Cake! - which will benefit St. Jude Children's Research Hospital - an incredible organization that's dedicated to treating children with cancer and other catastrophic diseases.
Let's Eat Cake! is a wedding cake competition, featuring Wedding Cake confections from all over the United States. Bakers will feature their cakes to be judged by Local and National Culinary Celebrities. In addition bakers will be asked to make enough cake to feed the general public, so that they may judge the cakes as well!
The goal is to raise $50,000 for this extremely worthy cause. (Donate here!) Please help spread the word about our goal, and our mission. If you can help financially, that'd be great, if you blogging about this right now…. your spreading the word - and that's all we can ask for!
Thank you!
Posted by Cakes By Shara at 4:46 AM 2 comments:
Monday, January 5, 2009
Cakes By Shara's Booth At Weddings the Bridal Show Saturday
Thank you to all the brides that came out to see us at "Weddings The Bridal Show." You will be notified if you were a winner of the bridal shower cupcakes we were offering.
Posted by Cakes By Shara at 1:53 PM 1 comment:
Labels: Cakes By Shara, Fondant Cake, Mad Hatter Cake, Nashville Weddings cakes, Rent A Cake, Weddings The Bridal Show 2009
Friday, January 2, 2009
Gray Photography! We Love Zach and Jody Gray.
Our Friends at Gray Photography took these lovely pictures of our cakes. Please visit them for your photography needs.
Posted by Cakes By Shara at 8:31 PM No comments:
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- Veteran defenceman and Duck captain (that sounds funny...) Scott Niedermayer who had previously hinted that he would call it a day after his last championship this spring is now realizing that pursuing one of the most brilliant careers ever may not be a bad idea after all.
- 110% last night was having a feast over La Presse's article featuring a vocal Saku Koivu managing expectations for the season to come; in short, the Captain is saying that the Cup is not a concern for now, nor is it realistic, it's about making the playoffs...just what I like to hear...
- The entire world is on the edge of its seat for Game 2 of the Junior Super Series that goes today between Canada and Russia...wake me up we rejoin the Westminster Dog Show in progress.
HF29 is still incapable of posting his famous TMS. His 56k modem seems to be failing him, as is evidenced by the distress smoke signals billowing out of his isolated retreat.


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($335, Rachel Comey, Neimanmarcus.com)Extra special hearts go to the double buckle motorcycle-inspired Barbaro. Order them (for me) now at Neiman Marcus.
Thursday, June 24, 2010
HUZZAH! Rachel Comey Pre-Fall Booties!
Ooh YAY! Direct your attention, will you, to Rachel Comey's Pre-Fall (isn't that the same thing as late-summer?) clunky, chunky heeled booties.
Posted by Tamar at 6:12 PM
Labels: ankle boots, booties, boots, Neiman Marcus, rachel comey
2 comments:
- The girl in the pearls. said...
These are gorgeous :) Please may I have some....
- Friday, June 25, 2010 3:56:00 PM
waw ,, i love it
- Friday, June 29, 2012 4:54:00 AM
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The control of excitable cell action potentials is central to animal behavior. We show that the egl‐19 gene plays a pivotal role in regulating muscle excitation and contraction in the nematode Caenorhabditis elegans and encodes the αl subunit of a homologue of vertebrate L‐type voltage‐activated Ca2+ channels. Semi‐dominant, gain‐of‐function mutations in egl‐19 cause myotonia: mutant muscle action potentials are prolonged and the relaxation delayed. Partial loss‐of‐function mutations cause slow muscle depolarization and feeble contraction. The most severe loss‐of‐function mutants lack muscle contraction and die as embryos. We localized two myotonic mutations in the sixth membrane‐spanning domain of the first repeat (IS6) region, which has been shown to be responsible for voltage‐dependent inactivation. A third myotonic mutation implicates IIIS4, a region involved in sensing plasma‐membrane voltage change, in the inactivation process.
Regulation of action potential duration is important for excitable cell function. For example, in neurons, the duration of action potentials at the synaptic terminal can affect the amount of transmitter released (Hochner et al., 1986; Spencer et al., 1989), whereas in vertebrate cardiac and gastrointestinal smooth muscles the duration of action potentials modulates the duration and strength of contractions (Noble, 1979; Huizinga, 1991). The physiological importance of regulating the duration of action potentials is exemplified by the human cardiac Long QT Syndrome (LQTS), in which the prolongation of the QT interval on electrocardiograms reflects a delay in the repolarization of ventricular myocytes. LQTS can manifest itself in ventricular fibrillation and syncopal episodes, ultimately leading to the death of young, otherwise healthy individuals (Schwartz et al., 1995).
In ventricular myocytes there are at least three major ionic currents contributing to the action potential: a regenerative Na+ current is responsible for fast depolarization, a Ca2+ current for the plateau phase and a K+ current for repolarization of the membrane. The activation and inactivation kinetics of each of these currents can affect the duration of the action potential. Indeed, mutations in a cardiac Na+ channel gene SCN5A and in a K+ channel gene HERG have been shown to be the causes of two distinct forms of congenital LQTS (Curran et al., 1995; Wang et al., 1995). Although many pharmacological and physiological studies have shown the importance of Ca2+ channels in shaping the cardiac action potential (Noble, 1979; Kass, 1995), mutations have not yet been found.
In general, the mechanisms regulating action potential duration are well conserved in metazoans. In recent years, it has been found that this mechanistic similarity extends to the molecular level. This conservation of molecular mechanisms has allowed physiologists to take advantage of the simplicity of invertebrates and their amenability to genetic approaches to help identify new molecules that are important for vertebrate excitable cell function. For example, the human HERG gene was cloned by its sequence homology to the Drosophila ether‐à‐go‐go (eag) gene (Warmke and Ganetzky, 1994). The eag gene was identified because of its effect on fruit fly neural and muscle functions when mutated (Zhong and Wu, 1991).
We are studying the control of muscle action potential duration at the molecular level by analyzing a simple neuromuscular pump, the pharynx of the nematode C.elegans. The pharynx, consisting of 20 myoepithelial muscle cells, 20 neurons and 22 other structural and secretory cells, is the feeding organ of the nematode. The muscles are arranged radially around a lumen, so that contraction opens the lumen and relaxation closes it. In the anterior pharynx, corpus muscle contraction serves to take in food (bacteria) suspended in liquid, whereas relaxation expels the liquid while trapping the food. In the posterior pharynx, terminal bulb muscle contraction rotates a grooved cuticular structure called the grinder that grinds bacteria and passes debris towards the intestine. Relaxation returns the grinder to its resting position. A pump is a cycle of nearly synchronous contraction and relaxation of the corpus and the terminal bulb (Albertson and Thomson, 1976; Avery and Horvitz, 1989).
The pharyngeal muscles are the most experimentally accessible excitable tissue in C.elegans. Pharyngeal electrical activity can be monitored in living animals by a simple extracellular recording called the electropharyngeogram (EPG; Raizen and Avery, 1994). The pharynx can be dissected from the rest of the animal, allowing pharmacological manipulation and direct measurement of muscle action potentials by intracellular recording (Davis et al., 1995). Similar to vertebrate cardiac and some smooth muscles, pharyngeal muscles can have myogenic activity (Avery and Horvitz, 1989). Contraction and relaxation are tightly correlated with the depolarization and repolarization phases of muscle action potentials. An action potential normally lasts about 150 ms (Raizen and Avery, 1994; Davis et al., 1995; Starich et al., 1996).
We have analyzed mutations that affect the duration of the pharyngeal muscle action potential. We find that egl‐19 encodes the α1 subunit of a putative voltage‐activated Ca2+ channel that is probably of the L type. Gain‐of‐function mutations in egl‐19 cause prolonged muscle action potentials and contractions (myotonic class), reduction‐of‐function mutations cause a reduced rate of depolarization and feeble contractions (flaccid class), and severe loss‐of‐function mutations lead to a complete loss of muscle contraction and thus lethality (lethal class). We have localized two myotonic mutations in the IS6 (the sixth membrane‐spanning domain in the first repeat) region and flanking residues. This region has been shown to control the rate of the voltage‐dependent inactivation between different types of Ca2+ channels (Zhang et al., 1994). We also found one myotonic mutation in IIIS4 (the fourth membrane‐spanning domain in the third repeat), a region that is a voltage sensor for channel activation (Catterall, 1995). Our results demonstrate the importance of a voltage‐activated Ca2+ channel in regulating the duration of action potentials. Furthermore, our mutant analysis suggests that the IS6 and IIIS4 regions play a role in L‐type Ca2+ channel inactivation.
Three classes of egl‐19 mutations affect muscle contraction and excitation
There are at least 26 mutant alleles of the egl‐19 gene (Table I, see Materials and methods for mutant isolation). These mutant alleles can be classified into three groups based on their genetic and phenotypic characteristics. The myotonic class of mutations causes a semi‐dominant excessive muscle contraction phenotype, which is a result of increased or misregulated gene activity. The flaccid class of mutations causes a recessive feeble muscle contraction phenotype which results from a partial reduction of gene function. The lethal mutations are recessive and cause a near‐complete block of embryonic muscle contraction as a consequence of a severely reduced or absent gene function. The inability of embryonic body muscles to contract leads to a distinctive embryonic lethal phenotype referred to as the Pat (Paralyzed, Arrested elongation at Two‐fold) phenotype (Williams and Waterston, 1994).
Myotonic mutations
Three mutations, n2368sd, ad695sd and ad952, are in the myotonic class. In an n2368sd mutant pharynx, the terminal bulb muscles, but not those of the corpus, often showed dramatically delayed relaxation (Figure 1A). To see whether the delay in relaxation was caused by delayed repolarization of muscle action potentials, we measured the electrical activities of the pharyngeal muscles by recording EPGs from intact worms. EPGs are analogous, in principle, to electrocardiograms or electroencephalograms used on humans. The EPG method provides a measurement of the capacitative current flows associated with changes in transmembrane potentials of pharyngeal muscle cells (Raizen and Avery, 1994). Thus, during a pump, fast depolarization of corpus and terminal bulb muscles in near‐synchrony together causes a group (usually a pair) of upward transients in EPG. Repolarization of the corpus produces a large downward transient followed by a smaller downward transient caused by the repolarization of the terminal bulb (Figure 1B; Raizen and Avery, 1994). EPGs showed that n2368sd mutant worms had terminal bulb muscle action potentials that were dramatically prolonged (one example is shown in Figure 1B). Delayed relaxation of the terminal bulb in n2368sd mutants is consistently associated with a delay in terminal bulb muscle repolarization (data not shown). This correlation between muscle relaxation and repolarization defects suggests that the muscle relaxation defect is caused at least in part by the repolarization defect. Consistent with their normal corpus muscle contractions, EPGs did not reveal a defect in corpus electrical activities in n2368sd mutant animals (Figure 1B). Nevertheless, n2368sd mutant animals also showed hypercontraction of several other muscles. They had a short and dumpy morphology (Figure 2A), possibly caused by excessive body muscle tone, and they were egg‐laying constitutive (Figure 2F), apparently because of excessive contraction of the egg‐laying muscles, as the frequency and duration of vulval muscle contractions were increased (data not shown). The n2368sd mutation is semi‐dominant. n2368sd/+ heterozygous animals had similar but weaker myotonic defects than those seen in homozygous mutant animals (data not shown).
ad695sd, like n2368sd, causes a semi‐dominant myotonic phenotype, although its defects are generally weaker than those of n2368sd animals (Table 1; Avery, 1993). Intracellular measurement of ad695sd terminal bulb muscle action potentials from two animals confirmed that ad695sd causes prolonged plateau phases (Figure 1C), as had been concluded earlier on the basis of EPGs (Raizen and Avery, 1994).
The ad952 mutation was isolated in a genetic screen for dominant suppressors of egl‐19(n582) (see Materials and methods). ad952 is tightly linked to n582 (within one map unit, see Materials and methods) and was shown by sequencing (see below) to be in egl‐19. n582 ad952 double mutant animals were essentially wild‐type in phenotype except that they were slightly dumpy, and the pharyngeal terminal bulb occasionally showed delayed relaxation and repolarization (Table I and data not shown). These phenotypes are similar to but much weaker than those seen in either n2368sd or ad695sd animals. n582 ad952/+ heterozygotes are wild‐type, unlike n2368sd/+ and ad695sd/+ heterozygotes. These results suggest that egl‐19(n582 ad952) is a weak gain‐of‐function mutant.
Flaccid mutations
egl‐19 flaccid mutants have feeble muscle contractions of both the corpus and the terminal bulb in the pharynx (Figure 1A). Their EPGs were normal with respect to the timing and amplitude of major signal peaks (Figure 1B). However, intracellular measurement of terminal bulb muscle action potentials in n582 (eight individuals, one typical action potential shown in Figure 1C) and ad995 (not shown) animals revealed that the rate of depolarization was reduced, suggesting that egl‐19 has a role in bringing about fast depolarization. In addition to the pharyngeal phenotype, flaccid mutants showed feeble body wall and egg‐laying muscle contractions. They tend to be long and thin, slow in movement and are egg‐laying defective (Figure 2C and H; Trent et al., 1983).
Lethal mutations
The lethal class of egl‐19 mutants has the Pat phenotype (Figure 2D and E). There was a dramatic reduction in the extent and frequency of embryonic body muscle contractions, and the most severe mutants did not contract at all (Williams and Waterston, 1994; data not shown). Embryonic body wall muscle contractions are apparently myogenic (Hall and Hedgecock, 1991; L.Avery, unpublished observations). Mutations in many genes important for muscle structure or function (for instance, those that encode myosin heavy chain A, vinculin, tropomyosin or troponin C) have the Pat phenotype (Williams and Waterston, 1994, and references therein), but no mutations in genes that affect only nervous system function have been found to have a Pat phenotype. The fact that the strongest alleles of egl‐19 eliminated embryonic muscle contraction suggests that egl‐19 affects embryonic muscle function directly rather than through the nervous system.
We conclude, based on mutant phenotypes, that the normal activity of egl‐19 is necessary for muscle depolarization and for regulating the duration of muscle action potentials.
Cloning of a C.elegans homolog of α1 subunits of vertebrate L‐type voltage‐activated Ca2+ channels
Genetic mapping data (Trent et al., 1983; Avery, 1993; Williams and Waterston, 1994; and Materials and methods) positioned egl‐19 in a small genetic interval between deb‐1 and dif‐1 on linkage group (LG) IV. This interval corresponds to a physical distance of approximately 150 kb in which we found a putative Ca2+ channel gene in independent experiments.
Specifically, by degenerate PCR (polymerase chain reaction), we cloned from C.elegans a cDNA fragment corresponding to the IIIS6–IVS6 region (from the sixth membrane‐spanning segment of the third repeat to the sixth membrane‐spanning segment of the fourth repeat) of known vertebrate voltage‐activated Ca2+ channel α1 subunits. We extended the sequence of the putative Ca2+ channel gene by screening a C.elegans cDNA library and by multiple steps of reverse transcriptase‐coupled polymerase chain reaction (RT‐PCR; see Materials and methods). We analyzed the presumptive ORF (open‐reading frame) of this putative Ca2+ channel gene (Figure 3A) by comparing its sequence to available protein sequences in databases and found that it is significantly more similar to known α1 subunits of L‐type channels than to other types of voltage‐activated Ca2+ channels (Figure 3B). Furthermore, this putative Ca2+ channel has most (19 out of 23) of the otherwise absolutely conserved amino acid residues found in domains IIIS5‐S6 and IVS5‐S6 of α1 subunits of known L‐type Ca2+ channels (Figure 3C). IIIS5–S6 and IVS5–S6 are two regions implicated in mediating channel sensitivity to 1,4‐dihydropyridines (Grabner et al., 1996). Dihydropyridine sensitivity is the defining characteristic of L‐type Ca2+ channels. In fact, pharyngeal muscles are sensitive to nifedipine, a dihydropyridine: nifedipine‐treated dissected pharynx showed feeble muscle contraction with an extended action potential (J.A.Dent, personal communication). Other muscles in C.elegans, for example body muscles, are also sensitive to L‐type Ca2+ channel blockers (L.Lobel and H.R.Horvitz, unpublished observations). Because of the high degree of sequence similarity, we believe that we have cloned the α1 subunit of a C.elegans L‐type Ca2+ channel.
We mapped the physical location of this Ca2+ channel gene to cosmid C48A7 by hybridizing the cloned cDNA fragment to a YAC grid, and subsequently to cosmids covered by the positive YAC clones (see Materials and methods). By comparing its sequence to the recently released C48A7 cosmid sequence (for a description of the C.elegans genomic project, see Wilson et al., 1994), we found that the Ca2+ channel gene lies completely within C48A7 (Figure 4).
egl‐19 encodes a voltage‐activated Ca2+ channel
C48A7 is between the genes deb‐1 and dif‐1 on LGIV, consistent with the location of the egl‐19 gene. We considered the possibility that egl‐19 is the Ca2+ channel gene, since egl‐19 mutant phenotypes are suggestive of defective muscle Ca2+ channels.
By germline transformation rescue experiments, we found that transgenic egl‐19(n582) mutant animals bearing the C48A7 transgene showed a nearly wild‐type phenotype with respect to pharyngeal, body and egg‐laying muscle contractions, suggesting that C48A7 contains sequences necessary for egl‐19 gene activity. By mapping the extent of the egl‐19 rescuing activity within C48A7, we found that the rescuing activity is co‐extensive with the Ca2+ channel gene (Figure 4).
To confirm that egl‐19 encodes the Ca2+ channel and to identify the molecular lesions in egl‐19 mutants, we determined the sequences of the entire coding regions of the Ca2+ channel in three egl‐19 mutants: the two myotonic mutants ad695sd and n2368sd, and the double mutant n582 ad952, which carries a flaccid mutation n582 and a myotonic mutation ad952. For each of these mutations we found a corresponding single‐base G:C→A:T transition in the coding sequence (Figure 3A). The ad695sd mutation changes alanine 906 in IIIS4 to a valine. In n2368sd, glycine 365 near the cytoplasmic end of IS6 is changed to an arginine. Two mutations were found in the n582 ad952 double mutant. Since the n582 ad952 double mutant was derived directly from an n582 single‐mutant animal, we were able to identify which of the two mutations found in the double mutant was n582 by determining the sequences of the two mutation sites in DNA isolated from n582 single‐mutant animals. The n582 mutation is in IIIS4 and changes arginine 899 to a histidine. The ad952 mutation changes serine 372 to a leucine close to the predicted intracellular end of IS6. The fact that there is a mutation in Ca2+ channel coding sequence corresponding to each of the four identified egl‐19 mutations, and that the egl‐19 rescuing activity is co‐extensive with the Ca2+ channel gene in the genomic sequence, argues strongly that the Ca2+ channel and egl‐19 are the same gene.
egl‐19 is expressed and functions in muscle cells
Although it is clear from mutant phenotypes that egl‐19 affects excitation and contraction of muscles, egl‐19 could nevertheless be influencing muscles indirectly, e.g. through the nervous system. To address the question of how egl‐19 affects muscle excitation, we localized its expression by assaying the expression of an egl‐19::GFP (green fluorescent protein, Chalfie et al., 1994) reporter (Figure 4). In transgenic animals carrying the fusion gene, an egl‐19::GFP fluorescent signal was first detected in body wall muscles in 1‐1/2‐fold embryos (Figure 5A and B), before the onset of embryonic muscle contraction. This result is consistent with the Pat phenotype seen in lethal mutants, suggesting a cell‐autonomous muscle defect. By the time of hatching, GFP fluorescence was found in pharyngeal muscles pm3, pm4, pm5 and pm7 (Figure 5C), in body wall muscles (Figure 5F) and in the anal depressor muscle (Figure 5D and E). The muscle expression pattern is again consistent with a muscle cell‐autonomous defect caused by mutations. We also found expression in the nervous system, including the pharyngeal neuron M4 and several neurons in the head, the ventral nerve cord and the pre‐anal ganglion (Figure 5C and E, and data not shown). This expression pattern suggests that egl‐19 may also function in neurons (see Discussion).
In the wild type, the HSN neurons are required for normal egg‐laying muscle contraction (Trent et al., 1983). We found that egg‐laying muscles in egl‐19(n2368sd) myotonic mutants contract even in the absence of HSN motor neurons (see Materials and methods). This observation suggests that egl‐19 acts in egg‐laying muscles to promote contraction. However, we could not detect GFP expression in egg‐laying muscles. The promoter fragment we used probably lacks elements for egg‐laying muscle expression (see Figure 4 and Materials and methods).
We have identified three classes of egl‐19 mutations. Mutations that belong to each of the classes have been isolated previously. We now know that egl‐19(n582), isolated by Trent et al. (1983), is a flaccid, partial loss‐of‐function allele, and eat‐12(ad695sd), isolated by Avery (1993), is a myotonic, gain‐of‐function allele. Six alleles of pat‐5 were isolated by Williams and Waterston (1994). These alleles are egl‐19 lethal, severe loss‐of‐function or null mutations (B.Williams, personal communication; see also Materials and methods). The phenotypes of these three classes of mutants considered together speak to the pivotal role that egl‐19 plays in muscle excitation and contraction.
Nematode pharyngeal muscles are similar to vertebrate cardiac and mammalian gastrointestinal smooth muscles that have slow‐wave‐type action potentials in several respects. For example, these types of muscles depolarize and contract cyclically; their action potentials have a long plateau phase; and the timing of contraction and relaxation is correlated to the depolarization and repolarization of their action potentials, respectively (Noble, 1979; Huizinga, 1991; Raizen and Avery, 1994; Davis et al., 1995). In vertebrates, L‐type voltage‐activated Ca2+ channels are known to be important in regulating the excitation and contraction of cardiac myocytes (Bean, 1989). Similarly, L‐type channels have been implicated in the slow‐wave‐type action potentials in mammalian gastrointestinal smooth muscles (Huizinga, 1991). Our results show that a putative L‐type voltage‐activated Ca2+ channel encoded by the egl‐19 gene serves a similar function in C.elegans pharyngeal muscles. These observations suggest that the nematode pharynx may be a good model system in which to study how varying Ca2+ channel activity may affect the physiology of muscles that have long action potentials.
egl‐19 encodes the major Ca2+ channel in C.elegans muscles
Most C.elegans muscles are sensitive to egl‐19 gene activity, including embryonic and post‐embryonic body wall muscle, pharyngeal muscle, egg‐laying muscle and possibly anal depressor and male spicule protractor muscle (Table 1; Figure 5). Therefore, EGL‐19 is the α1 subunit of a major muscle voltage‐activated Ca2+ channel in C.elegans. To date, only one other voltage‐activated Ca2+ channel αl subunit has been genetically characterized (Schafer and Kenyon, 1995). In contrast to egl‐19, unc‐2 encodes an α1 subunit more similar to non‐L than L‐type voltage‐activated Ca2+ channels. Reduction of unc‐2 function leads to an egg‐laying constitutive phenotype, opposite to that of egl‐19 reduction‐of‐function. Furthermore, unc‐2 primarily affects neural function (Schafer and Kenyon, 1995; Schafer et al., 1996). unc‐2, however, does have a functional role in body muscles. unc‐2 mutants are sluggish in movement, and based on mosaic studies, Schafer and Kenyon (1995) concluded that this phenotype is caused by the loss of unc‐2 function in body muscles. Thus, although our data suggest that egl‐19 is functionally the major voltage‐activated Ca2+ channel in muscles, other Ca2+ channels may also contribute to muscle excitation and contraction.
The expression pattern of an egl‐19::GFP reporter indicates that egl‐19 is also expressed in many neurons. We do not know the functional significance of the nervous system expression, since most of the defects of egl‐19 mutants are consistent with defects in muscle function exclusively. However, muscle defects would mask most nervous system defects. One exception is that ad695sd and n2368sd animals have a dauer‐formation defect (Daf‐d). A dauer larva is an enduring, dispersal form of larva that is of a developmental stage equivalent to an L3. Although some dauers did develop from starved ad695sd or n2368sd mutant L2 larvae, many of these dauer animals appeared to be incompletely modified, as they were less resistant to SDS treatment than wild‐type dauers (data not shown). Normal dauers have modified cuticles which render them resistant to harsh environmental conditions (Riddle, 1988). Since the formation of dauers is largely dependent on neural function (Riddle, 1988), and no mutations affecting only muscle function have been reported to cause a dauer‐formation defect, this Daf‐d phenotype suggests that egl‐19 may also function in the nervous system.
Control of pharyngeal muscle repolarization
Although the EGL‐19 channel clearly affects the action potentials of both the corpus and terminal bulb muscles, based on the expression pattern and the loss‐of‐function phenotype, gain‐of‐function mutations cause a myotonic phenotype only in the terminal bulb. This observation suggests that different mechanisms are involved in the control of action potential duration in these two parts of the pharynx. This notion is also supported by the fact that the corpus always repolarizes before the terminal bulb. It seems possible that the corpus has an intrinsic ability to repolarize earlier than the terminal bulb. To achieve a well‐regulated and fast repolarization, other ion channels must participate in the repolarization process. For instance, mutations in a glutamate‐gated Cl− channel, encoded by the avr‐15 gene, cause a delay in muscle repolarization. AVR‐15 is apparently part of the pharyngeal muscle receptor for the neurotransmission from M3 inhibitory motor neurons, which directly affect the repolarization of the corpus but not the terminal bulb (Dent et al., 1997). Therefore, M3 activity could preferentially hasten corpus repolarization. However, M3 transmission alone can not entirely account for the difference, since the terminal bulb still repolarizes later than the corpus when M3 transmission is blocked (Raizen and Avery, 1994). Another channel likely to play a role in the repolarization process is the nematode negative spike K+ channel, originally characterized by Byerly and Masuda (1979) in the Ascaris pharynx (see also Davis et al., 1995). A difference in the expression or regulation of activity of this and perhaps other channels could account for the different repolarization timing between parts of the pharynx.
Myotonic mutations in egl‐19
The myotonic phenotype of egl‐19 mutants is well explained by slower inactivation kinetics of the muscle Ca2+ current, since that Ca2+ current is likely to be the major inward current during muscle depolarization. When inactivation of the EGL‐19 channel is retarded, the muscle membrane potential would be held depolarized, and the resulting sustained Ca2+ influx would prolong muscle contraction. L‐type voltage‐activated Ca2+ channels are known to inactivate by two mechanisms, voltage‐ and calcium‐dependent. These two means of inactivation are thought to operate independently (Hadley and Lederer, 1991).
By assaying the inactivation kinetics of chimeric channels (made between an α1A and a marine ray homolog of the α1E class) in Xenopus oocytes, Zhang et al. (1994) found that a region of the proteins that includes IS6 and flanking residues is responsible for the different rates of voltage‐dependent inactivation in voltage‐activated Ca2+ channels. These authors also suggested that this type of inactivation is reminiscent of the C‐type (slow) inactivation observed in voltage‐activated K+ channels, revealed after the removal of N‐type (fast) inactivation by deletion of residues that form the tethered plug to the channel pore (Hoshi et al., 1991). Recently, a mutation (N434A) in IS6 of the rat μ1 Na+ channel protein has also been found to affect the slow inactivation of the channel (Wang and Wang, 1997). Two of egl‐19 myotonic mutations, n2368sd and ad952, are in the IS6 region. We believe that the myotonic phenotype observed in these mutants is caused by a defective voltage‐dependent inactivation of the EGL‐19 channel. Thus our data extend the importance of the IS6 region in controlling channel inactivation to L‐type channels.
The third myotonic mutation, ad695sd, is in IIIS4. S4 segments are thought to be the voltage sensors for voltage‐sensitive channels (Catterall, 1995). Indeed, an R→H mutation in IIIS4 causes the reduced rate of depolarization observed in n582 mutants, as would be expected if this mutation affected the voltage sensor of the channel. It is possible that the A→V mutation in IIIS4 found in ad695sd animals also affects the activation of the channel. However, we have not observed significantly slowed‐down depolarization kinetics in ad695sd, in contrast to the dramatically prolonged plateau phases in mutant muscle action potentials. Although the kinetics of channel activation can affect the kinetics of inactivation, it seems unlikely that ad695sd mutant channels inactivate slowly simply as a consequence of slow activation kinetics. Thus, the mutation apparently uncouples the role of IIIS4 in channel activation from its role in inactivation. A similar phenomenon has been reported for mutations in the IVS4 segment in vertebrate skeletal muscle Na+ channels. R1448C and R1448H mutations in the α subunit of the human skeletal muscle voltage‐activated Na+ channel gene SCN4A cause paramyotonia congenita. When the biophysical properties of the channels were assayed in a cell line, it was found that the mutations have only a small effect on activation but dramatically slow inactivation (Chahine et al., 1994).
The two myotonic mutations in the IS6 region of the EGL‐19 channel may also affect the activation phase of muscle action potentials. First, n2368sd has a cold‐sensitive Pat phenotype (Table I and Materials and methods). This phenotype is apparently caused by a reduction in gene function, since it is recessive to the wild‐type allele and is not complemented by other Pat alleles. How the n2368sd mutation imparts this phenotype is not clear. A possibility is that the G→R mutation in IS6 alters the structure of the protein only slightly at high temperatures (∼20°C) to affect channel inactivation, whereas the mutation destablizes the protein dramatically at low temperatures (∼12°C) to impair channel formation or function. Second, we have noticed in intracellular records that n582 ad952 double mutants have consistently faster depolarization kinetics than n582 single mutants (unpublished observations). Since we do not have the ad952 mutation in isolation, we do not know if the suppression between n582 and ad952 is mutual. It should be interesting to assay the detailed biophysical properties of each of these mutant channel proteins in a simpler and more electrophysiologically accessible system, e.g. Xenopus oocytes.
In conclusion, our analysis of myotonic mutants in C.elegans has implicated a voltage‐activated Ca2+ channel in regulating the duration of muscle action potentials. Our in vivo analysis suggests that mutations in IS6 and IIIS4 regions of the α1 subunit can dramatically affect Ca2+ channel inactivation. We envisage that continued genetic analysis of C.elegans pharyngeal excitation will further our understanding of the molecular mechanisms that underlie the control of action potential duration in excitable cells.
Materials and methods
General methods
Worm culture, handling and genetic manipulation followed the methods described by Sulston and Hodgkin (1988). Except for cold‐sensitivity experiments, all worms were maintained at 20°C. The wild type was the N2 strain of the Bristol variety of C.elegans. Methods used for analyzing pharyngeal behavior and electrical activity have been described elsewhere: timing diagrams by Avery (1993); EPGs by Raizen and Avery (1994); and intracellular recordings by Davis et al. (1995).
Mutant isolation and genetic characterization
All mutagenesis was carried out with ethyl methanesulfonate (EMS), which causes primarily G:C→A:T transitions (Anderson, 1995). Mutants isolated were backcrossed at least twice against N2 or bli‐6(sc16) unc‐24(e138am) animals.
n2368sd. Mutagenized egl‐1(n986dm) V males were crossed with unc‐79(e1068) ced‐4(n1162) III; rol‐4(sc8) unc‐76(e911) V; lon‐2(e678) xol‐1(y70) X hermaphrodites. Most viable progeny from this cross should be hermaphrodites with the genotype unc‐79 ced‐4/+; rol‐4 unc‐76/egl‐1; lon‐2 xol‐1/+ since the xol‐1 mutation kills the male progeny which do not have a wild‐type copy of the X chromosome. These hermaphrodites have an egg‐laying defective phenotype, since the egl‐1 mutation dominantly causes death of the HSN motor neurons (Trent et al., 1983). Two F1 animals showed normal egg‐laying among 14 000 scored. One carried n2368sd and the other a new ced‐4 allele (Ellis and Horvitz, 1986). We found that the HSNs were still absent in n2368sd; egl‐1(n986dm) double‐mutant animals (data not shown) but the egg‐laying muscles were now able to contract.
n2368sd was mapped to LGIV between unc‐24 and bli‐6 by two‐ and three‐factor crosses. n2368sd is tightly linked to ad695sd, since in Unc non‐Bli recombinants segregated from bli‐6(sc16) egl‐19(ad695sd) unc‐24(e138am)/egl‐19(n2368sd) either ad695sd (10/12) or n2368sd (2/12) was present. This result suggests that n2368sd and ad695sd are no more than 0.5 map units apart. We found no wild‐type recombinants among 12 800 progeny segregated from n2368sd/n582 heterozygote mothers. This result indicates that the n2368sd mutation can be no more than 0.05 map units (roughly 50 kb) from n582. egl‐19(n582) was isolated by Trent et al. (1983) for its egg‐laying defective phenotype.
Loss‐of‐function mutations. New loss‐of‐function mutations were isolated by non‐complementation screens against n582. Either mutagenized N2 males were crossed with bli‐6(sc16) egl‐19(n582) unc‐24(e138am) hermaphrodites, or mutagenized N2 hermaphrodites were crossed with bli‐6(sc16) egl‐19(n582) unc‐24(e138am)/+ males. Cross progeny were scored for the Egl‐19 phenotype. Twelve independent new alleles were isolated from a total of 6 900 haploid genomes screened. Each was within 3 map units of n582.
Since five of these 12 egl‐19 mutants had a Pat phenotype similar to that of pat‐5 mutants and since pat‐5 maps close to egl‐19 (Williams and Waterston, 1994), we tested whether egl‐19 and pat‐5 are the same gene. None of four pat‐5 alleles (st553, st556, st569 and st571) complemented egl‐19(n582). These results and the fact that pat‐5(st556) was rescued by the same cosmid that rescued egl‐19 (see below) suggest that pat‐5 is egl‐19. We believe that Pat is the null phenotype of egl‐19 because the most severe Pat alleles behave like a chromosomal deletion (a bona fide null) in heterozygotes. For example, n582/nDf41 animals did not show a more severe phenotype than n582/ad1004 animals.
egl‐19(n582) was mapped previously between unc‐8 and dpy‐20, near the position +3.4 on chromosome IV (Trent et al., 1983). To map further egl‐19(n582), we isolated recombinants from a strain of the genotype unc‐44(e362) deb‐1(st555) unc‐24(e138) / egl‐19(n582). Eleven out of 24 Unc‐24 non‐Deb recombinants carried egl‐19(n582). This result places egl‐19 approximately half‐way between deb‐1 (position +3.3) and unc‐24 (position +3.5), which are approximately 800 kb apart on the physical map (Coulson et al., 1995). Since nDf41 deletes egl‐19 and has its right breakpoint near or within dif‐1 (position +3.4; J.Ahringer, personal communication), egl‐19 must be close to or to the left of dif‐1, which is ∼400 kb from deb‐1. Together, these results place egl‐19 in an approximately 150 kb interval to the left of dif‐1.
cis‐acting suppressors of n2368sd and ad695sd. We used a genetic cis‐trans test to establish that the myotonic mutations are alleles of egl‐19. We introduced egl‐19(null)‐like mutations in cis to n2368sd (Table 1). The rationale is that if a gain‐of‐function and a null mutation are in the same gene, the null mutation should occlude the expression of the gain‐of‐function if these two mutations exist in the cis configuration, but not when they are in the trans configuration. We made egl‐19(null) mutations in cis to n2368sd by non‐complementation screens. We crossed mutagenized n2368sd hermaphrodites with bli‐6(sc16) egl‐19(n582) unc‐24(e138am)/+ males. One Egl mutant (allele ad1023) was isolated among 750 haploid genomes screened. egl‐19 mutations were introduced onto the ad695sd mutant chromosome by a similar non‐complementation scheme. Among 3 600 mutagenized haploid genomes screened, three alleles (ad1000, ad1002 and ad1021) were isolated as Egl mutants. Each of these four new mutations was tightly linked (within 3 map units) to the original semi‐dominant allele, n2368sd or ad695sd, respectively. All of these double mutants were wild type as heterozygotes. In contrast, both n2368sd and ad695sd, when heterozygous with an egl‐19(null) mutant chromosome, caused a myotonic phenotype. All five double‐mutants were Pat as homozygotes. Thus, egl‐19(null) mutations are cis‐dominant trans‐recessive suppressors of n2368sd and ad695sd. We conclude that n2368sd and ad695sd are mutations of egl‐19.
ad952. We screened for non‐Egl F1 progeny of mutagenized egl‐19(n582) hermaphrodites. One such mutant (ad952 n582) was found among 15 000 haploid genomes. ad952 was tightly linked to n582 and mapped between bli‐6 and unc‐24, which are ∼1 map unit apart.
Tests for cold sensitivity
Gravid hermaphrodites were kept on petri plates in a box placed in a circulating cold water bath in order to maintain a tightly controlled temperature for the embryos. n2368sd embryos were nearly 100% Pat at 12°C. Only a small fraction were affected when grown at 15°C. We found no cold sensitivity for ad695sd, n582 or N2 animals.
cDNA cloning and sequencing
General cloning methods followed those described by Sambrook et al. (1989). For sequencing, we used an ABI 377 automatic sequencer. For sequence management, we used the GCG Wisconsin Package.
Degenerate PCR primers were designed based on the conserved sequences in the IIIS6 and IVS6 domains of the α1 subunits of vertebrate L‐type voltage‐activated Ca2+ channel genes (primer sequences available upon request). A 953 bp PCR product was amplified from worm cDNA and cloned (pCAC1‐953). The cDNA was localized in the genome by hybridizing CAC1‐953 to a YAC poly‐grid, poly 1 (kindly provided by R.Waterston; Coulson et al., 1988). Only two overlapping YAC clones, Y51C8 and Y49F12, showed positive hybridization. Three cosmid clones, C48A7, B0496, and K11C12 (kindly provided by Alan Coulson), together span the overlap of the YACs. By Southern hybridization, we found that CAC1‐953 is contained entirely in C48A7, partially in B0496, but not in K11C12. C48A7 has been placed between two genes deb‐1 and dif‐1 on the physical map (Coulson et al., 1995).
Using pCAC1‐953 as a probe, we screened a mixed‐stage worm cDNA library (Barstead and Waterston, 1989) to isolate a 2.8 kb cDNA clone (pCE12A). We then designed PCR primers based on the sequences in pCE12A (TTATTTTGGAATGTAACGATAACGAAACCT) and in the genomic sequence of cosmid C48A7 (provided by the C.elegans sequence consortium) corresponding to the first predicted transmembrane domain (IS6) of the Ca2+ channel gene (CTTTGCCTCTCCTTGAATAATCCCATCCGA) to RT‐PCR more 5′cDNA sequence from a cDNA mix made from mixed‐stage poly‐A selected total RNA. A 3 kb PCR product (e12‐RT1) was cloned and sequenced. To obtain further 5′ sequence, we used the Genefinder program (P.Green, unpublished; cited in Eeckman and Durbin, 1995) to help us identify the possible initiating ATG from the genomic sequence. We used a primer corresponding to the sequence immediately 5′ to the ATG (GGAGTGCGCCGACACTGCTCGATCGTGA) and another primer corresponding to the 5′ portion of e12‐RT1 (TACTCCTGATACAAGACGAAGCGGT) to PCR‐amplify a 0.7 kb product from a cDNA mix. The sequence of the PCR product was determined by direct sequencing. pCE12A lacked the poly‐A signal sequence, suggesting incomplete 3′ extension. A cDNA clone (yk53d3), isolated and kindly given to us by Yuji Kohara, overlaps in sequence with pCE12A and extends 0.5 kb further 3′ to include the AATAAA poly‐A signal. When assembled together, these four pieces would make one 6.2 kb cDNA. The authenticity of this presumptive transcript was verified by RT‐PCR using the most 5′ and 3′ primers. A single product of the predicted size was amplified. Based on Northern blots using CAC1‐953 as the probe, the full‐length transcript should be ∼7 kb long. Nevertheless, we believe that the entire ORF is in our sequence, since there are stop codons in all three frames preceding the first in‐frame ATG and following the last predicted amino acid.
Isolation of genomic lambda phage clones and germline transformation
We used CAC1‐953 as a probe to screen a C.elegans genomic lambda FIXII phage library (Stratagene). The sizes and extents of the inserts in phage clones that we isolated were determined by restriction pattern analysis and by direct sequencing of the ends using primers corresponding to the multiple cloning site of the vector. We were able to map these clones precisely, since the sequence of cosmid C48A7 became available during the course of our analysis.
The ability of these phage clones and of C48A7 to rescue egl‐19 mutants was determined by germline transformation experiments (Mello et al., 1992). The genomic clone was co‐injected into egl‐19(n582) animals with pRAK3, which contains a dominant rol‐6 marker (Mello et al., 1992; Davis et al., 1995). Transgenic lines that stably expressed the marker were established and observed for Egl‐19 defects, including abnormal egg‐laying, body morphology, movement and pharyngeal pumping. For C48A7, two of eight lines showed rescue; for F2 and F3 co‐injection, five of five; for F2 alone, six of eight; for E3, zero of three; for H1, none of nine; for E2, nine of nine.
Some unusual phenomena associated with the F2 and E2 transgenes are worth noting. We have observed occasional terminal bulb muscle relaxation defects in some transgenic animals carrying the F2 clone, while their sisters (also transgenic) appeared to be wild type. This variability may reflect the fact that F2 contains an egl‐19 gene truncated at the 3′ end (see Figure 4). Rescue of egl‐19(n582) by E2 transgenes was incomplete. In particular, the egg‐laying defective phenotype was only slightly improved in all E2 transgenic animals, whereas pharyngeal and body muscle phenotypes were fully rescued. This result suggests that E2 may lack important regulatory elements for proper expression in egg‐laying muscles. This hypothesis could explain why the egl‐19::GFP reporter was not expressed in egg‐laying muscles, as the egl‐19 regulatory sequence in the reporter was from E2 (see below and Figure 4).
One transgene carrying C48A7 also rescued an embryonic lethal allele, st556 (Williams and Waterston, 1994). Males of the genotype egl‐19(st556)/unc‐82(e1323) unc‐24(e138am) IV (a gift from B.Williams) were crossed with the transgenic line egl‐19(n582) IV; adEx1058[egl‐19+ rol‐6(d)]. Viable and sometimes fully wild‐type (except for the Rol phenotype of the co‐injected rol‐6(d) gene) animals were segregated of the genotype egl‐19(st556) IV; adEx1058[egl‐19(+) rol‐6(d)].
Mutation detection
Genomic DNA was prepared from animals homozygous for the respective mutations. Segments of the egl‐19 gene were amplified by PCR. The sequences of the entire coding regions and intron‐exon boundaries from ad695sd, n2368sd and n582 ad952 animals were determined by direct sequencing of PCR products. The sequences were compared to the genomic sequence of the C48A7 cosmid made available by the C.elegans Genome Consortium. Wherever a suspected mutation was found, the sequencing was repeated at least once more to eliminate potential artifacts. The sequences of the oligonucleotides used for amplification and/or sequencing of the exons are available upon request.
egl‐19::GFP fusion
A translational fusion of egl‐19 and GFP was made by inserting a 4.7 kb fragment of egl‐19 into the multiple cloning site in the pPD95.70 GFP expression vector (A.Fire, J.Ahnn, G.Seydoux and S.Xu, personal communication), which also contains a nuclear localization signal. The egl‐19 fragment was prepared from phage E2 (see Figure 4). E2 DNA was digested with BcgI. The open ends were blunted by Klenow enzyme. E2 was then digested again with SalI, which cuts in the vector. The 4.7 kb fragment was gel‐purified. The pPD95.70 vector was prepared by digestion with SmaI and SalI followed by gel purification. Ligation of these two fragments resulted in fusion of the egl‐19 translation frame to that of the GFP. The fusion construct (peat‐12::sGFP‐NLS) was cloned and checked by restriction digests and used to germline transform lin‐15(n765ts) animals along with DA#735 (a plasmid containing lin‐15; Huang et al., 1994) as a co‐injection marker. Transmitting lines that segregated wild‐type animals were established and the expression of GFP was observed in wild‐type animals by fluorescence microscopy with an FITC filter set. We had 10 independent lines expressing egl‐19::GFP, and they looked essentially the same with respect to where the signal was detected. The strength of the signal, however, varied widely among them. We therefore concentrated our further analysis on the line that had the strongest expression. We present (in Figure 5) a view of the pharynx in a dauer animal, because the expression pattern of egl‐19::GFP in the pharynx is most easily photographed in dauers, as the pharynx is flat and compact.
We thank the C.elegans Genome Consortium for cosmid sequences, A.Coulson for cosmids, Y.Kohara for a partial cDNA clone of egl‐19, A.Fire for GFP expression vectors, R.Waterston for YAC filters, B.Williams for pat‐5 mutant strain, and K.S.Liu and J.A.Dent for sharing unpublished results. Some strains used in mapping experiments were supplied by the Caenorhabditis Genetics Center, which is supported by the National Institute of Health National Center for Research Resources. We also thank M.W.Davis, J.A.Dent, R.Joho, F.Katz, D.Raizen and R.Zwaal for their constructive comments on the manuscript. This research was supported by United States Public Health Service research grants HL46154 to L.Avery and GM24663 to H.R.Horvitz. H.R.H. is an Investigator at the Howard Hughes Medical Institute.
- Copyright © 1997 European Molecular Biology Organization


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Your baby is cranky. Their sleep is all over the place. You've ruled out ill health, hunger, thirst, dirty nappy, too hot, too cold... You think it's overtiredness, but even after a good nap or first thing in the morning they're still clingy and easily upset. What can explain it?
It seems that some crankiness seems to be part of normal, healthy baby development. Think of it like morning sickness. It's not much fun, but on the upside it's usually a sign that everything's on track and the baby's developing normally. There's nothing you have to do about it, except to do your best to comfort your baby and show you care.
Here are some of the most popular explanations I've found for baby crankiness.
The Period of Purple Crying (0-12 weeks)
It is normal for babies to be placid in the first 2 weeks after birth, then round week 3 they start to cry more and more. The crying just happens for no reason and can be impossible to soothe. One way of thinking about this is as 'The Period of Purple Crying'. 'Purple' is an acronym that stands for Peaks at 6-8 weeks, Unexpected, Resists soothing (no matter what you try), Pain-like face, Long-lasting (up to 5 hours a day in some cases), and Evening (because it seems to occur more in the late afternoon / evening). Here is a video:
It is so common for 0-12 week old babies to have an inconsolable period in the evening, that it can be known as the 'witching hour' or 'arsenic hour'. There are some great tips on how to realistically approach soothing a baby at this age here on the Period of Purple Crying website.
Colic is not a condition so much as a name that is applied to babies who cry a lot. As in, colic literally means that the baby cries a lot and we don't know why. A colicky baby cries a lot. We're talking hours of endless crying, and no amount of soothing seems to help. Colic is defined as 3 hours a day, at least 3 days a week, for more than 3 weeks. This is a bit of an arbitrary division that researchers use when classifying babies as colicky or non-colicky. No one has found a definitive cure or explanation for really persistent criers or colicky babies. Some claim to have some success with baby-wearing, probiotics, switching formula or certain types of bottles for bottle-fed bubs etc., so you can always try these things and see if they help.
Growth Spurts
It is not so much that growth spurts are associated with crankiness, but that they are associated with a trying time for parents, particularly in the early days, because at least the 3 week and 6 week growth spurts seem to throw a baby's sleep out of whack, and breastfed bubs feed round the clock during a growth spurt to get enough milk and bring up the milk supply, making it a demanding time for mum in particular. There are typically growth spurts at 7-10 days, 2-3 weeks, 4-6 weeks, 3 months, 4 months, 6 months, and 9 months.
Wonder Weeks
I just noticed in the last day or so Bethany seems to have suddenly got... well... smarter. It's hard to explain. It's as though when I look at her I see that the cogs in her brain are turning that little bit faster. Turns out there is a '55 week' wonder week, after which babies have a much more refined sense of cause and effect.
Wonder weeks are mental growth spurts that all babies allegedly go through over the course of the first year. This idea comes from some Dutch researchers, Frans Plooij and Hetty van de Rijt who set out their theory in the book The Wonder Weeks.
What is so useful for parents about the wonder weeks concept is the idea that when the mental growth spurt happens, most babies go through a period of being inexplicably cranky. They are easily upset. They sleep poorly. And some of the 'weeks' go on much longer than a week. Knowing this clingy, cranky behaviour can be normal and will soon pass can really help you be patient with your baby during these trying periods.
You can look up more detail about when the wonder weeks are supposed to occur and what is developing on the Wonder Weeks website. If you want a quick, useful reference for where your baby is at, there is an iPhone app about wonder weeks.
Some people speculate there is no such thing as teething, although most parents will swear there is. Once the period of colic ends, unexplained fussiness and whinging gets labelled 'teething' whether there's any teeth in sight or not. All babies seem to go through a very 'mouthy', dribbly period regardless of whether teeth appear then or much later. My baby was particularly clingy and whingy as her first teeth were coming through (6-9 months), but whether that was due to the teeth, to separation anxiety, or something else, I have no idea. Perhaps some teeth hurt and others don't. Perhaps it varies between babies. My baby has been getting her first molar this last week, and apart from being a bit more distressed when she woke at night, there were no behavioural changes that I noticed at all - and as mentioned above it was apparently also the right timing for a wonder week, so who knows? I've tried a few things for teething, such as toys for mouthing and Bonjela, but didn't notice her take to them. I didn't try an amber necklace and the pain hasn't been noticeably bad enough that I would give her Panadol.
That said, some babies seem to get quite marked symptoms like a rash or diarrhoea when a tooth is coming through. Sometimes teething seems to really cause a lot of pain - other times it is just a bit annoying, so that your baby can be distracted, but when he gets tired and things go quiet he gets really upset.
Sleep Regressions
Babies often sleep better, then worse again, then better, then worse. Sometimes this is explained as having a 'sleep regression'. There is talk particularly of sleep regressions at 4 months and 9 months. This may, however, just be caused by wonder weeks. See this discussion.
Breastfed babies often start all-night feeding binges round 4-6 months because they are so much more alert that they are too distracted to feed enough during the day.
It is not a novel idea that some babies react poorly to some foods. We are all on the lookout for major reactions the first time we offer foods like nuts, eggs, shellfish, dairy, gluten etc. Babies with reflux can become worse with foods that are too runny, or if they don't get enough of a chance to burp upright.
But an intolerance to foods or food additives can also be more subtle and ongoing. I only really looked into this when I thought Bethany had a mild rash in response to grapes sprayed with sulfur dioxide. I found this website on the FAILSAFE method really useful for understanding what food additives there are, how to watch out for them / avoid them, and how to do an elimination diet to try and identify whether any of the additives are having an effect.
There are also culprits like caffeine to watch out for. You probably aren't giving your baby coffee or chocolate, but if you are breastfeeding, it can go through your breastmilk.
Contrary to popular belief, there seems to be increasing evidence that sugar is not responsible for making kids hyper. Our perception that it does may be in part a placebo effect, both on the parents (who perceive their child's normal exuberance as sugar related) and kids (who learn that hyperactivity is expected of them after sugar). But it also may be to do with all the other things that normally come along with sugar - such as excessive amount of articificial colourings and flavourings, and caffeine.
Tuesday, July 19, 2011
Explanations for Your Cranky Baby
Posted by livingindarwin at 10:18 PM
1 comment:
Thanks GNP. I have this book and have found it very helpful as we approach the 19 week leap. Flo will be all smiles then burst into tears for no apparent reason ... which is associated behaviour with this leap! Phew. It helps to get a bit of a heads up, provided you don't then project the behaviour onto your baby :) Natalie.ReplyDelete
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R&B pop queen Rihanna recently launched the eighth single from her '07 album Good Girl Gone Bad. Impressed yet? Here is a quarter century of other milestones to dazzle you.
7 Singles in the Top Ten 10
Born In The USA (1984)
Bruce Springsteen The Boss' rock opus hit the top 10 like a star-spangled cherry bomb.
5 Singles at No. 1
Bad (1987)
Michael Jackson Thriller was the MJ whammy everyone remembers, but Bad was a singular event all its own.
7 Singles in the Top 5
Rhythm Nation 1814 (1989)
Janet Jackson Yes, it's a concept album about social injustice. But it's also packed with dance hits.
6 Singles in the Top 40
Come On Over (1997)
Shania Twain With Top 40 hits and 11 singles on the country charts, it's still the crossover album to beat.
4 Singles in the Top 10
Breakaway (2004)
Kelly Clarkson An epic 103 weeks on the Billboard 200 broke Celine Dion's 1998 record.


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I gotta admit sometimes trolls are a little entertaining.
Well, the word "troll" is also used for promiscuous gay men who troll around the video-coin booths at porn shop trolling for other men.
You can imagine my adjustment.
Changing the word "cyber" on me was really annoying too.
I think the word "troll" is used to dismiss people too often. I generally reserve it for people who are saying extremely rude/absurd/abusive things just to get attention.
In my happy-place they are all just frustrated performance artists.
Although the internet term "troll" is considered to be about trolls under bridges, the fishing definition is just as apt.
From the WordWeb dictionary:
(n.) a fisherman's lure that is used in trolling
(v.) to angle (fish) with a hook and line drawn through the water
I'd thought that "troll" in the internet sense actually did derive from the angling sense – the person doing it was trailing a baited hook to see who would react to them.
Pardon me for interrupting, but what's going on? Battalion Commander dead in Fort Bragg Shooting
I hope you are OK.
So what. I like what she does now………. Everyone is free to do whatever he wants. ……..by the way this is old news.
Didaktylos: Yes, that's correct, that was how the term was used on Usenet on the alt.usenet.kooks newsgroup, where the Bob Allisat and Pierre Salinger "Hook, Line & Sinker" awards were given out: http://www.coloneljake.com/AUK/AAD/
Now it has come to mean anyone who is intentionally seeking conflict, though often still with the connotation that it's an insincerely expressed position.


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3480 North Lake Shore Drive
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From the Director:
"Goals include the continued development of socialization and independence in a nurturing environment; and integration of family and school within a Jewish context. Additionally the Nursery Programs help the children form a positive Jewish self-image; and provide stimuli and opportunities for growth and development in social, cognitive, intellectual, and physical skills." - Taken from website
"The curriculum is grounded on the idea that children learn through play, and the environment is carefully planned to foster discovery, exploration and learning. We work to give our children a positive self-image, high self-esteem and a strong Jewish identity. The Gan Shalom staff consists of professionals specializing in early childhood education. There is a special 15% discount for all Temple Sholom of Chicago members in good standing."
- Taken from website
"A typical morning would find the children engaged in free play; singing together and sharing their experiences during group time; trying a science experiment; enjoying snack time; following through on assigned classroom responsibilities for the day; creating art projects related to a theme or upcoming holiday. Children receive all readiness skills to assure a successful transition to kindergarten. The Jewish calendar cycle and holidays play an important role in the classroom. Shabbat is celebrated each Friday, with parents baking challah on a rotating schedule, with the children. Special holiday celebrations with families are scheduled throughout the school year, both during class hours and at other times. Hebrew is introduced in the classroom informally through songs and games so that the children become thoroughly familiar and at ease with the language of their religion."
- Taken from website


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The University of California system has said "enough" to the Nature Publishing Group, one of the leading commercial scientific publishers, over a big proposed jump in the cost of the group's journals.
On Tuesday, a letter went out to all of the university's faculty members from the California Digital Library, which negotiates the system's deals with publishers, and the University Committee on Library and Scholarly Communication. The letter said that Nature proposed to raise the cost of California's license for its journals by 400 percent next year. If the publisher won't negotiate, the letter said, the system may have to take "more drastic actions" with the help of the faculty. Those actions could include suspending subscriptions to all of the Nature Group journals the California system buys access to67 in all, including Nature.
The pressure does not stop there. The letter said that faculty would also organize "a systemwide boycott" of Nature's journals if the publisher does not relent. The voluntary boycott would "strongly encourage" researchers not to contribute papers to those journals or review manuscripts for them. It would urge them to resign from Nature's editorial boards and to encourage similar "sympathy actions" among colleagues outside the University of California system.
Along with its letter, the California Digital Library included a fact sheet with systemwide statistics for 2010 about the university's online journal subscriptions. The system subscribes to almost 8,000 journals online, at an average cost of between $3,000 and $7,000 per journal, depending on the publication and the field. The current average cost for the Nature group's journals is $4,465; under the 2011 pricing scheme, that would rise to more than $17,000 per journal, according to the California Digital Library.


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Friday, June 22, 2012
Photo shoot
I have been crazy busy this week preparing for my first product photo shoot! I am so excited and incredibly thankful for this amazing opportunity! Wish me luck today & I will hopefully share photos with you soon!
Posted by Susan Crabtree at 7:40 AM
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Skye Sports Bra
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Be pretty in pink with this medium support sports bra. Designed with our performance LJ Excel™ fabric, featuring a higher neckline for both a fashionable and flattering fit, the front and back keyholes allow for extra breathability. We love the way the Hyper Pink hue adds that feminine touch to your workout look.
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The, usually minor, change in viscosity that occurs when an electric field is applied to some polar liquids.
Subjects: Chemistry.
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One important feature, I enjoy during my travel is the differences, just travel for an hour within a country(Italy) and how much differences you can experience.Especially this time, traveling with my brother who lived almost 10 years in North America and showing him the Europe for the first time, it was quite an experience. He freaked out seeing the houses as old as his American towns still happily floating in the Mediterranean sea in Venice, Navigating using the small city map in Rome, well! here the roads not exactly meet at right angles to each other, you are bound to get lost and use that as an excuse to stop an handsome Italian man to ask for directions, (thanks to the confusing roads of Rome). Small scooters rushing in the old stone paved roads of Florance, the church chandelier which inspired Galileo in Pisa. Not just the stones of colosseum, every stone in Italy was narrating the history of THE Romans. Not just this, the types of food, the amount of cheese used (still people are slim?,) the hotel rooms still use the classical keys (No electronic card), no street name in the cross roads (street names are mostly written on the buildings in the cross road),the size of the shower rooms etc. etc. every small things put my brother in surprise. Now that I am bit used to Europe, when I can I see something ' very different'.....very soon, I have to now run to two embassies to process my two visas to two countries in 'Middle East' (are they exactly in Middle East? ...may be..) and then join my friend Alcidio for lunch and hear about his last two months adventures in Africa. Thanks to the differences which still survive in the tough hands of Globalization.
Nothing exists except atoms and empty space; everything else is opinion -- Democritus(460-370 BC)
October 31, 2007
October 30, 2007
Be a traveler or tourist, Italy have nice things to offer, check out the photos of two Indian tourists lost in the beauty of Italy. Trip to Italy with my brother.
October 20, 2007
Welcome to Europe!
Can you guess how long it takes to go from Paris to its airport? With the train strikes and the rugby final match adding fuel to the fire, it took me 9 hours, from door to door to go and receive my brother from the airport! Well! I didn't have much time to complaint about it, now that my brother is with me and we are doing a stormy (you can all call it as "quick and dirty" or "a look and feel") tour in Italy. yeah! that's the first country he wanted to see in Europe, even before Paris, that too the whole country in just three days! Inspite of all I am having a great time, introducing my brother to my favorite Europe! I will update soon about the "look and feel" trip feels like.
October 08, 2007
Lot of things!
This Saturday was nuit blanche and the French's fate deciding rugby match. Had a nice Italian dinner party at my boss's house with his very first and latest students, both latest finished and latest ongoing (Anton and Me) watching the rugby match for the very first time in my life and that too I was able to catch my host country winning a very tough game. Humm! for the very first time in my life some events got repeated, Last year I had a stroll in my favorite square with Anton to see it lighted in blue for nuit blanche, this year out of mere coincidence we just repeated that after the dinner, this year the same square was lighted with lamps with fire. Check out the photos, you will not deny that it was taken in the romantic city, Paris. Well! this year too the road was filled with drunken people, even some drunken monkeys went into a museum (which is open over night for Nuit Blanche) and punched some million dollar worth Claude Monet's painting it seems! crazy!
October 02, 2007
Heureusement que dans vie ...
Heureusement que dans vie certaines choses refusent de changer (Fortunately somethings in life doesn't change).. Like me trying to learn a language. I joined my French class again for the third time, i successfully failed and was happily taken in level one by the Paris municipality for adult education. It was my first day in a cute little école élémentaire (elementary school), sitting in the tiny chairs and benches I started my French classes all over again. Like our parents get us new bags and pencils to encourage us to go to school, Anton sent me this French (Canadian) song, very meaningful, its about how fast things are changing from generation to generation.. I am sure one day in my life I can speak French!
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Lady Gaga, Robin Hood Foundation, and Hetrick-Martin
Lady Gaga and the Robin Hood Foundation are partnering to donate $1 million to support organizations that help disconnected youth in NYC. As a Hetrick Martin staffer, I am not allowed to vote so VOTE FOR ME. You can cast your vote for Hetrick-Martin to win the donation at http://bit.ly/GagaRobinHood!
OR you can follow the links below:
Link to our Facebook Page: http://www.facebook.com/HetrickMartin
Link to Lady Gaga's Page: http://www.facebook.com/ladygaga?v=app_158498217539308
Link to our Causes Page: http://www.causes.com/causes/56396-the-hetrick-martin-institute
Link to the Contest Page: http://ladygaga.robinhood.org/
Note: Voting can only occur through Lady Gaga's page and through a link on our Causes page.
I appreciate everyone donating their time to support us :)
Wednesday, April 27, 2011
She's GAGA over HMI!
Posted by Erica at 1:00:00 PM
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Alexander Ludwig - Kids' Choice Awards 2013 Red Carpet
Alexander Ludwig high-fives some fans on his way up to the stage at the 2013 Kids' Choice Awards held at the USC Galen Center on Saturday (March 23) in Los Angeles.
The 20-year-old actor joined co-stars Willow Shields and Amandla Stenberg and accepted the blimp for Favorite Movie for The Hunger Games.
PHOTOS: Check out the latest pics of Alexander Ludwig
"Just accepted the KCA for best movie. #thehungergames So honored to have such devoted fans. Thanks to all that voted! #love," Alex tweeted from backstage.
10+ pics inside…
Posted to: 2013 Kids Choice Awards, Alexander Ludwig


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Blind MLB pitcher
They don't know because they don't ask, and they don't ask because they don't want to stir any bad memories. And even if they were to, they probably wouldn't believe Sandoval. Because the idea of a blind baseball player really is that difficult to fathom.
Never will he forget the day: Feb. 4, 2006. And the scene: At a restaurant where a drunken man, angry with the bouncer who booted him, returned with the shotgun. And the noise: A blast, loud and furious. And the feeling: Panic and fear and chaos as three buckshot pellets lodged in his right eye. And the sight.
"Black," he said. "Nothing but black."
It's still black, permanent darkness out of the right side of his head, his depth perception nonexistent and his left eye forever tasked with transmitting the world into his head. And because his left eye remains good – better than it's ever been, in fact – he is still here, still playing baseball, still trying one more time to reach the major leagues after years on the cusp as a right-handed pitcher, standing on the rubber and staring in with that good eye.
The story of how Juan Sandoval came to be a Tampa Bay Ray is one of this spring's best, and even if he is a long shot to make the team now, this is no gimmick, no publicity stunt. It is about an organization that looks high and low for talent, a city in the Dominican Republic that produces it in spades and a player getting one final chance to fulfill what he's certain is his birthright: wearing a major league uniform for the first time, at 32 years old.
read more: http://sports.yahoo.com/news/blind-in-one-eye--right-hander-juan-sandoval-still-sees-bright-future-with-rays-070949850.html


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#10: The Boer War, Britain's Vietnam, a guest post by the late historical fiction author T. D. (Tim) Griggs about the last days of empire, comparing Britain in the late 1800s with America in the following century.
#9: A visual preview of the winter season: Downton Abbey readalike edition. In November 2012, Downton Abbey's influence on historical novels was emerging. This post contained blurbs for a dozen upcoming novels of interest to fans of the show.
#8: Bestselling historical novels from 2009 looked at the top historical fiction sellers from that year, according to Publishers Weekly. Unfortunately PW is longer producing these annual compilations.
#7: Bestselling historical novels from 2011, the same thing for 2011.
#6: An updated post, which has almost no content in itself, other than a link to my historical fiction picks from BEA 2011. It's odd this one ranks so highly, but it's probably because of the keywords used in it.
#5: A Puritan Maiden's Diary: The Early American Primary Source That Wasn't, an essay about discovering how a diary supposedly written by a teenage girl in 17th-century Rhode Island was actually a piece of historical fiction written in the late 19th century. I reported the error to the Library of Congress, and it's since been taken out of their archive of primary sources recommended to teachers for use in the classroom.
#4: A report on Sarita Mandanna's novel Tiger Hills, which looked at this expansive saga set in Coorg in southern India between 1878 and WWII.
#3: The power of point of view, author Victoria Wilcox's guest essay about how she used different points of view to show the many facets of Doc Holliday's character, even ones that don't agree with our own moral values.
#2: My 1000th blog post, another milestone post, which previews 10 new and upcoming historical novels that caught my interest in October 2014.
#1: The Billy Sunday Snowstorm, author Barbara J. Taylor's guest post, written to mark the 100th anniversary (3/1/2014) of the time when charismatic preacher Billy Sunday was snowed in at his tabernacle overnight with many Scranton residents during an unexpected blizzard. The snowstorm was featured in Barbara's debut novel Sing in the Morning, Cry at Night. This post has been read nearly 29,000 times to date.


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I was checking out Aiden's A Dribble of Ink blog this morning and saw that he found an online test to see what kind of reader you are. I took it and got this:
|What Kind of Reader Are You?
Your Result: Dedicated Reader
You are always trying to find the time to get back to your book. You are convinced that the world would be a much better place if only everyone read more.
|Obsessive-Compulsive Bookworm
|Literate Good Citizen
|Book Snob
|Fad Reader
|What Kind of Reader Are You?
Quiz Created on GoToQuiz
That sounds about right I think. ;-)


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The Rutgers Quizzing Club is an informal group that meets every other week, and everyone is invited to participate - even people not associated with Rutgers. The quiz masters are chosen by turn and the teams are decided impromptu. It is basically an exercise in fun. Yesterday was my turn to be quiz master. The questions are here.
P.S. If you want to participate in future session(s), drop me a line. (Yeah, this is an unofficial promotion!)
Thursday, April 06, 2006
Posted by Sailesh Ganesh at 1:51 AM
Labels: quizzing
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Welcome to Rug Tub - Memphis Rug Cleaning....by PSH Cleaning Services
Rug Tub, a division of PSH Cleaning Services, is a professional, full-service Oriental, Persian and area rug cleaning company serving the West Tennessee area, including Memphis, East Memphis, Germantown, Collierville, Bartlett, Lakeland, Midtown, Arlington. PSH Cleaning has offered premiere area rug cleaning in Memphis since 2005.
At Rug Tub, we take pride in our reputation, experience, education, systems, and guarantee. Rug Tub is certified by the IICRC with multiple cleaning technician certifications, which are the highest available in the carpet cleaning industry. Rug Tub has long held the standard for Memphis rug cleaning.
You can depend on Rug Tub's parent company, PSH Cleaning Services, for services like: carpet cleaning, upholstery cleaning, carpet protectant and upholster protectant, tile and grout cleaning and sealing, natural stone cleaning and sealing, marble polishing and sealing, and urine damage treatment.
5640 Summer Ave Suite 1
Memphis, Tennessee 38134


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Wet DreamsFavorites [+]
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Running Time: 103 minutes
Released: 10/2012
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The Huffington Post recently interviewed a group of men about their opinions on masculinity. What emerges is a view of (at least hegemonic) masculinity as restrictive and unachievable. As one man states,
"I think a lot of us pretend to be quite different than we really are."
And they all appear to reach the same conclusion – that we'd all be much better off without it:
"In an ideal world..masculinity wouldn't mean anything"
Do you agree? Does the world need masculinity and femininity? In your ideal world, what would masculinity mean?


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Well... that makes five. There are now FIVE completed Requiem trilogies. Fifteen novels in this crazy world where people can become dragons.
Five years ago, on a cold Sunday morning, I stepped into a bookstore in downtown Toronto. I had a notebook, a pen, and an MP3 player with Mozart's Requiem playing. I sat at a little coffeeshop inside the bookstore, listened to the music, and began to write whatever came to mind. That turned into Blood of Requiem, the first novel in the series. I had no idea it would even get ONE sequel, let alone grow into this massive world spanning fifteen books and counting.
Requiem now includes the following trilogies: Dawn of Dragons, Song of Dragons, Dragonlore, The Dragon War, and Requiem for Dragons. So far, you've all downloaded half a million copies of these books. Wow.
Between these trilogies, we got to explore some other worlds together. We spent six novels in Moth, and we spent three novels with the Alien Hunters. But we keep coming back to these old dragon friends.
I'm honored that, five years into this, you're still reading. I really do have the best job--and the best readers--in the world.
Dragons Rising, which was released today, is not the last Requiem novel. This world will continue to grow. l can't wait to see where we'll fly together next.


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Hi everyone!
Please check out my WordPress blog: Clutter Cubed!
It's the blog of a young woman just trying to clean out all the junk in her life and the struggle against clutter and compulsive hoarding. A self-help blog for those with clutter problems.
That's where all my posts will be.
Thank you! ^_^


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I was over at angeltreats a few weeks ago and spotted this post and I just had to go make me into a SuperHero. I made Mr T into one too.
I think I'd need to get a lot slimmer and Mr T would need to get a lot more buff to fit into these costumes but I'm really rocking that cape.
Check out The Hero Factory to make your own.