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Holidays are a wonderful time, but they can also be stressful. Lots of rich food, endless activities, late nights, sugar galore... it's no wonder many of us suffer burnout by Jan. 2. But with some simple tips, families can avoid holiday overload and maximize the fun of the season.
1. Save the schedule. Many kids are sensitive to schedule changes, especially when it comes to sleeping. Holiday celebrations are likely to go much smoother if kids can maintain regular nap times and bedtimes. I know it's challenging to plan sleep around holiday activities, Christmas parties and New Year's Eve celebrations. Certainly a hectic day here and there won't wreck the nightly routine. But pint-sized Grinches start to rear their heads if too many nap times and bedtimes are missed. It's worth a little planning and sacrifice to let the little ones' slumber.
2. Spare the stomach. It's hard to resist the plates of goodies popping up everywhere, and the dinner table loaded with special holiday food. Again, a schedule can help here. Kids (and parents!) are less likely to overeat the junk food if there are still regular, healthy meals and snacks throughout the day. Even a small handful of raw almonds or an apple can tide you over and impart the willpower to pass the cookie tray.
3. Mind the medical conditions. This may sound like a no-brainer, but in the hustle of the holidays it can be easy to forget medications or overlook allergies. Especially at a party where you may not know all the food ingredients. Enlist kids' help to remember when it's time to take pills or use asthma inhalers. Make sure relatives and hostesses know if there's a food allergy issue so everyone can be on the lookout.
4. Enlist the elves. I have trouble delegating daily duties especially during Christmas time. But I'm slowly learning that I don't have to do everything on my own! My kids are itching to help whenever I let them. Children can be great at manning the tape dispenser for gift wrapping, putting bows on gifts, decorating the Christmas tree, loading the car before a trip, keeping the house clean, and even baking and delivering treats to neighbors. Why fly solo for the entire Christmas season? Share the joy!
5. Out with the old. Here's another great way to involve kids in the holidays: Participate in a toy clean-out. We ask our kids to choose a boxful of their gently loved toys to give to other little kids who don't have presents for Christmas. We talk about how blessed we are as we sort the toys and imagine how happy some little child will be. Together, we take the toys to Goodwill. Not only do the kids feel good about helping others, they clear some room for new Christmas gifts.
6. Travel savvy. If your holiday plans include traveling be sure to talk to kids about the plans. Let them know where you're going — maybe show them on a map so they have an idea of the distance. Help them plan activities for the car or plane so they don't get bored within the first five minutes. Be sure to hydrate, take regular stops to stretch legs and use the restroom, and have light snacks to (hopefully!) minimize motion sickness.
7. Ease into electronics. If kids receive an iPad or video game console for Christmas, they're going to want to play it nonstop. That's probably not catastrophic in the short term. But it's possible to get headaches and jitters from hours of straight electronic entertainment. The American Academy of Pediatrics recommends no more than two hours of screen time daily. Parents may consider breaking up playtime on the new iPad into distinct chunks (say, 30 minutes at a time), followed by a mandated hour or two of some other activity. Who knows... kids may get so involved in building a snowman or sledding in the backyard that they give the electronics a rest for an entire afternoon.
8. Caution is crucial! The U.S. Centers for Disease Control (CDC) urges caution during the holidays. Many injuries occur at this time of year. Keep power cords well away from kids and high-traffic areas. The CDC warns especially about candles — they are extremely dangerous around kids, pets, and anything flammable. If you love candles at Christmas time, it's a great idea to put fresh batteries in smoke detectors and carbon monoxide alarms. Parents of small children must be especially cautious about choking hazards (like candy, Christmas ornaments, toys, wrapping paper, etc.) and potential injuries. We learned one year just how quickly a curious baby can pull a huge Christmas tree over on top of himself. Talk to older kids about safety issues and make sure everyone stays vigilant.
9, Healthy, happy holidays. As folks congregate for parties, germs get passed around along with the egg nog. Make sure to wash hands often with soap and warm water and avoid touching the face. Help kids learn to cough or sneeze into their upper sleeve so they don't spread more germs. And be sure to dress properly for the frosty temperature — infants and older adults are especially susceptible to health problems caused by cold exposure. Use multiple layers of tightly-woven clothing and avoid staying too long in extreme temperatures.
Happy Holidays from our family to yours! May your days be merry and bright, full of fun, and as stress-free as possible.
How do you keep your cool and avoid holiday overload? Share thoughts in the comments section below!
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We have previously developed bioengineered three-dimensional internal anal sphincter (IAS) rings from circular smooth muscle cells isolated from rabbit and human IAS. We provide proof of concept that bioengineered mouse IAS rings are neovascularized upon implantation into mice of the same strain and maintain concentric smooth muscle alignment, phenotype, and IAS functionality. Rings were bioengineered by using smooth muscle cells from the IAS of C57BL/6J mice. Bioengineered mouse IAS rings were implanted subcutaneously on the dorsum of C57BL/6J mice along with a microosmotic pump delivering fibroblast growth factor-2. The mice remained healthy during the period of implantation, showing no external signs of rejection. Mice were killed 28 days postsurgery and implanted IAS rings were harvested. IAS rings showed muscle attachment, neovascularization, healthy color, and no external signs of infection or inflammation. Assessment of force generation on harvested IAS rings showed the following: 1) spontaneous basal tone was generated in the absence of external stimulation; 2) basal tone was relaxed by vasoactive intestinal peptide, nitric oxide donor, and nifedipine; 3) acetylcholine and phorbol dibutyrate elicited rapid-rising, dose-dependent, sustained contractions repeatedly over 30 min without signs of muscle fatigue; and 4) magnitudes of potassium chloride-induced contractions were 100% of peak maximal agonist-induced contractions. Our preliminary results confirm the proof of concept that bioengineered rings are neovascularized upon implantation. Harvested rings maintain smooth muscle alignment and phenotype. Our physiological studies confirm that implanted rings maintain 1) overall IAS physiology and develop basal tone, 2) integrity of membrane ionic characteristics, and 3) integrity of membrane associated intracellular signaling transduction pathways for contraction and relaxation by responding to cholinergic, nitrergic, and VIP-ergic stimulation. IAS smooth muscle tissue could thus be bioengineered for the purpose of implantation to serve as a potential graft therapy for dysfunctional internal anal sphincter in fecal incontinence.
- tissue engineering
- smooth muscle contraction
fecal incontinence (FI) is an embarrassing disorder that can have overwhelming social and psychological impacts on an individual. The prevalence of FI has been estimated to fall within the range of 2.2–15%, with the numbers increasing with the age of the population. Although the etiology of FI can be multifactorial, it is well established that FI is secondary to obstetric trauma, idiopathic sphincteric degeneration and/or dysfunction, anorectal surgeries, and irritable bowel syndromes (1, 2).
The internal anal sphincter (IAS) contributes to 70% of the high anal canal pressure required to maintain rectoanal continence. This high pressure, dubbed as the basal sphincteric tone, arises from the state of contracture of the circular smooth muscle of IAS (29). The source of this high basal tone is thought to be primarily myogenic (25) and is modulated by neurohumoral components. Passage of colonic content through the anal canal during defecation is accompanied by relaxation of the IAS smooth muscle, with an associated giant migrating contraction propagating caudally in the colon. IAS dysfunction can largely be attributed to its weakened mechanical efficiency at the smooth muscle level or pudendal neuropathies (16, 24). Loss of basal sphincteric tone due to idiopathic degeneration of smooth muscle cells, stretching of elastic tissue and heightened collagen/fibril content as a result of aging, surgical trauma, or gut resections all lead to FI (19).
Conventional treatment options like dietary manipulations and antidiarrheal drugs can significantly reduce FI associated with chronic diarrhea. Other therapeutic efforts in FI focused on sacral nerve modulation, electrically stimulated gracilis muscle transposition, or biofeedback rehabilitation of the pelvic floor are not documented to be effective. Although these treatment options might provide relief in FI symptoms in individuals with structurally intact sphincters, sphincteric damage and dysfunction can only be corrected by invasive surgical options. Injection of bulking agents like glutaraldehyde cross-linked collagen, polytetrafluoroethylene, polydimethylsiloxane, etc., to improve sphincter function is accompanied by the danger of allergy and the possibility of migration of these particles into the lymphatic circulation, triggering a more severe allergic or foreign body response. Artificial anal sphincters could be implanted but present with the potential of device failure, infection of the electrodes, or severe foreign body reactions (11, 20, 29, 30).
The lack of an ideal therapeutic option for a disabling state such as FI has prompted our laboratory to develop a bioengineering solution focused on providing a positive remedy to FI due to degenerated IAS. We have previously bioengineered functional three-dimensional IAS constructs from mice, rabbits, and humans out of isolated circular smooth muscle cells from their respective internal anal sphincters (14, 27). Recently, we bioengineered mouse IAS rings for the purpose of implantation. We provide proof of concept that implantation of bioengineered IAS constructs into mice (of the same strain as the cell source) display healthy vascularization, physiological functionality before and after implantation, and no signs of rejection or inflammation.
Intestinal tissue engineering has shown exciting prospects in short bowel syndrome with and without the use of scaffolds (18). Skeletal muscle and cardiac pressure generating constructs have been developed before and tested in vitro for physiological function, but a successful implantation and comparisons of functionality have previously not been worked on (10, 15).
In this study, IAS rings were bioengineered in a parallel manner from circular smooth muscle cells from the mouse internal anal sphincter. One ring was tested for physiological functionality and another was implanted into an animal of the same strain. Harvested postimplant IAS displayed neovascularization. The functional studies on the postimplant harvested constructs focused on the following: 1) maintenance of overall IAS physiology; 2) maintenance of membrane ionic characteristics; 3) integrity of membrane receptors and intracellular signaling pathways for contraction and relaxation of IAS. Implanted rings showed similar functional behavior to bioengineered IAS rings preimplantation in their ability to 1) generate spontaneous basal tone; 2) contract in response to potassium chloride, acetylcholine, and phorbol dibutyrate; and 3) relax in response to vasoactive intestinal peptide (VIP), nifedipine, and nitric oxide donor sodium nitroprusside (SNP).
Our experiments provide proof of concept that bioengineered IAS constructs can be implanted in the animals without rejection, are neovascularized, and can remain physiologically functional. This implantable bioengineered IAS tissue could be used as autologous graft tissue for degenerated IAS. Combined with an increased knowledge of molecular mechanisms underlying smooth muscle function, this opens out doors to novel therapeutic options for severe FI attributed to IAS dysfunction.
C57BL/6J female, specific pathogen-free mice (8 wk old) were obtained from Jackson Laboratory (Bar Harbor, ME) and maintained under temperature-, humidity-, and light-controlled conditions. The study conformed to the Guidelines for the Care and Use of Laboratory Animals established by the University Committee on Use and Care of Animals at the University of Michigan, and protocols were approved by that committee (no. 09714).
All tissue culture reagents, including Dulbecco's modified Eagle's medium (DMEM), trypsin, media supplements like l-glutamine, and antibiotics, were purchased from Invitrogen (Carlsbad, CA). Sylgard, i.e., Poly(dimethylsiloxane), was purchased from World Precision Instruments (Sarasota, FL). Acetylcholine, VIP, phorbol dibutyrate, nifedipine, and SNP were obtained from Sigma (St. Louis, MO).
Isolation and culture of IAS cells.
IAS was identified and cultured as previously described (3, 5, 14, 27). Briefly, C57BL/6J mice were euthanized by carbon dioxide inhalation. Mouse sigmoid colon and rectum along with the anal canal was removed by sharp dissection. The tissue was kept moist in Hanks' balanced salt solution (HBSS; Hyclone, Logan, UT) containing antibiotics and antimycotics at 4°C. The fecal content was cleaned and it was slipped onto a moist Pasteur pipette. Adherent fat, connective tissue, striated muscle, blood vessels, and serosa were stripped off and cleaned with HBSS-wetted Kimwipe (Kimberly-Clark, Neenah, WI). The tissue was cut along its longitudinal axis and the mucosa was stripped off. The IAS was identified as a 2- to 3-mm strip of circular smooth muscle at the distal end of the rectum under a dissection microscope. IAS circular smooth muscle was removed by sharp dissection. Isolated IAS was then minced and digested twice with 0.1% type II collagenase (Worthington Biochemicals, Lakewood, NJ) and 0.1% 0.2 mg/ml DNAse (Roche Applied Sciences, Indianapolis, IN) at 32°C for 1 h each and filtered through a 500-μm Nytex (Tetko) mesh. The circular smooth muscle cells were collected by centrifuging and resuspended and washed three times to remove excess collagenase. Dispersed circular smooth muscle cells were then plated with growth media (GM: DMEM + 10% fetal bovine serum + 1.5% antibiotics/antimycotics + 1.5% l-glutamine) on to regular tissue culture T-75 flasks. Plates were incubated at 37°C with 5% CO2, with an accompanied medium change every 48 h until the cells were ready to be used.
Preparation of culture dishes.
The 35-mm tissue culture plates were coated with 0.8 ml Sylgard and allowed to cure for 2–3 days. Sylgard molds (5-mm diameter) were fixed to the center of the plate with insect pins to define luminal space. The plates were sterilized with ethanol and under UV light for 30–45 min. Thrombin (400 μl; 10.5 U/ml in GM) was mixed with 200 μl of fibrinogen (20 mg/ml) on the Sylgard-coated culture dish. In 30 min, the thrombin and fibrinogen polymerized to form a loose fibrin gel.
Bioengineering three-dimensional mouse IAS constructs.
Circular smooth muscle cells from the IAS were lifted off from their culture surface with 3 ml 0.25% trypsin-EDTA. The harvested cells were centrifuged and counted with a hemocytometer. Cell suspension concentration was adjusted to 1 × 105 cells/ml of GM; 2 ml of this cell suspension was plated on top of the loose fibrin gel in each prepared culture plate. As the cells proliferated in the fibrin gel, they started contracting the gel, toward the center of the plate where the post was affixed. GM was switched out to differentiation medium (DM: DMEM 365 ml, Media-199 100 ml, horse serum 35 ml, Pen/Strep 5 ml). This aided smooth muscle differentiation. The cells migrated in the fibrin gel, arranged themselves concentrically around the Sylgard post, and resulted in a thick three-dimensional IAS ring held together by cellular ECM in 4–6 days. The DM was replenished every 2 days. At this point, they were ready to be tested for physiological functionality and implantation.
Surgical procedure.
Surgical procedure for implantation of bioengineered IAS constructs has been described by Hashish et al. (13). Briefly, the Alzet model 1004 microosmotics pumps (Durect, Cupertino, CA) were primed per manufacturer's instructions. Sufficient volume of human recombinant fibroblast growth factor-2 (ProSpec-Tany TechnoGene) was loaded into pumps to allow for continuous infusion over a 21-day period. A sterile polyethylene catheter (0.76 mm, Durect) was attached. This pump-catheter complex was kept in sterile 0.9% saline at room temperature for 48 h prior to implantation.
The mice were anesthetized using an intraperitoneal injection of 100 mg/kg of ketamine and 5 mg/kg xylazine. After confirmation of adequate anesthesia, the surgical area on the back of the mouse was clipped of hair and prepped with chlorhexidine.
A 2-cm transverse skin incision was made in the upper back of the mouse (midscapular incision). A subcutaneous pocket was created caudally, which was made large enough to fit the pump and catheter comfortably. The bioengineered IAS ring was inserted near the tip of the catheter, ∼0.5 cm away from the skin incision. A 6-0 Prolene suture was placed loosely around the ring to mark its position. The catheter tip was then cut to length so that the tip was directed toward the implanted ring. The catheter was then secured to the underlying tissue by a single 4-0 Prolene suture.
The skin incision was then closed with 4-0 Vicryl sutures. The animals were returned to an approved facility for recovery in standard fashion.
Ring harvest.
Mice were euthanized 25–28 days postimplantation. The area of the previous incision was reentered and the rings were smoothly removed from the surrounding tissue after localization of the Prolene suture. The harvested construct was placed in PBS and was assessed immediately for physiological functionality as per protocols outlined below.
Following measurement of physiological functionality (below), half of the harvested ring tissue was placed in zinc-formalin for fixation, and the other half was placed in Tissue-Tek OCT compound (Ted Pella) and frozen in liquid nitrogen. Slides were then prepared and sectioned by the Tissue Core at the University of Michigan Comprehensive Cancer Center. Hematoxylin and eosin (H&E) and Masson's trichrome staining were used for morphological assessment. Immunofluorescence assays were carried out for α-smooth muscle actin (F3777, Sigma), smooth muscle-specific heavy caldesmon (c-4562, Sigma), and endothelial-specific antigen von Willebrand's factor (factor VIII; sc-14014, Santa Cruz Biotechnology). Slides were visualized by an Olympus FV500 fluorescence microscope in the University of Michigan Microscope and Image Analysis Laboratory.
Western blot analysis.
A small portion of the postimplant bioengineered IAS ring and intact mouse IAS tissue were minced, sonicated, and lysed in RIPA buffer. Equal amounts of protein were separated on SDS-PAGE and electrophoretically transferred onto polyvinylidene difluoride membranes. Membranes were blocked with 5% nonfat dry milk for 1 h and incubated with the appropriate dilution of the following primary antibodies in 3% nonfat dry milk in PBS with Tween-20 (PBST): c-Kit (sc-168, Santa Cruz Biotechnology) and prolyl-4-hydroxylase (MAB2073 Millipore). β-Actin (A5441, Sigma) was used as a loading control. Unbound primary antibody was removed by washing with PBST. The membrane was then incubated for 1 h with the appropriate dilution of horseradish peroxidase-conjugated secondary antibody. PBST washes were used to remove unbound secondary antibody. Blots were visualized with chemiluminescence.
Measurement of physiological functionality.
Protocols for measurement of force generation were adapted from previous work on three-dimensional IAS constructs (14, 27). The tissue bath in the force transducer set up consisted of a Sylgard coated 35-mm culture plate with a stainless steel pin fixed to the center of the plate. The plate held 4 ml of basal serum-free DMEM and was placed on a heated aluminum platform that maintained the temperature at 37°C ± 1°C. Before any measurements were made, noise was measured by recording tension with air and warm bath fluid without hooking the bioengineered tissue. The mean noise due to bath fluid was subtracted from total tension recordings with the tissue. Passive tension was obtained by hooking on the tissue with no stretch in zero calcium + 50 mM EGTA at the end of the day. This passive tension value was subtracted from total tension recordings with the tissue to obtain active tension. Values of force reported are only active tension values.
The IAS ring was separated from the Sylgard mold using forceps, and one end was anchored to the fixed pin in the tissue bath. The other end of the ring was hooked on to the movable measuring arm of an isometric force transducer (Harvard Apparatus, Holliston, MA). Stock solutions of the contractile and relaxant agonists were 40 times more concentrated than the required final concentration so that a uniform application of 100 μl resulted in the desired final concentrations in a 4-ml static bath. Bath diffusion times were assessed by use of methylene blue and averaged up to 44.3 ± 2.8 s for complete uniform diffusion of 100 μl in 4 ml. Basal DMEM soaking the ring was changed every 30 min, or at the end of a set of experiments, whichever was earlier.
Testing protocol.
Bioengineered rings or harvested postimplant rings were hooked on to the force transducer with no additional stretch. Basal tone was established spontaneously with no external stimulation or stretch over time as shown in Fig. 7. A 20% stretch was applied on the constructs by a micromanipulator and this remained the length at which all tests were carried out. The baseline of force established by the rings at this length was arbitrarily set to zero, to observe effects of drugs on the constructs. The rings were allowed to develop stable baseline before any contractile agonist/relaxant peptide was added to the tissue bath. Maintenance of membrane characteristics were studied by treatment with 30 mM potassium chloride. Receptor integrity and intracellular signaling mechanism preservation was studied by observing 1) relaxation of the basal tone of the IAS ring by addition of 1 μM VIP, 10 μM nifedipine, and 10 μM SNP; 2) cholinergic stimulation with 1 μM acetylcholine; 3) and direct stimulation of signal pathway intermediary PKC with 1 μM phorbol dibutyrate.
Data analysis.
Data was acquired by LabScribe2 (iWorx, Dover, NH) at a frequency of 0.02 Hz. GraphPad Prism 5.01 for Windows (GraphPad Software, San Diego, CA; www.graphpad.com) was used for all further data analysis. Second-order Savitsky-Golay smoothing was applied on raw data. Values were expressed as means and SE of 4–10 experiments. Contraction and relaxation were quantified as a percentage of each individual construct's basal tone. KCl-induced contraction was expressed as a percentage of peak maximal contraction induced by acetylcholine.
Bioengineering IAS rings.
Mouse IAS cells isolated from primary cultures were expanded separately on T-75 culture flasks, and 200,000 cells were seeded onto loose thrombin polymerized fibrin gels. The cells in the fibrin gel started extending, 4 h postseeding (Fig. 1B). Over the next 24 h, the cells proliferated and oriented themselves concentrically around the Sylgard post (Fig. 1C). In 3 days, the fibrin gel shrank and contracted toward the center of the 35-mm culture dish, resulting in the formation of a three-dimensional mouse IAS ring (Fig. 1D). The different time points in the formation of the bioengineered ring are depicted in Fig. 1. The resulting rings had an inner diameter of 5 mm and an approximate thickness and width of 1–2 mm. They were stable in culture for over 35 days. IAS rings were bioengineered in a parallel manner. One ring was implanted, whereas the other was assessed for physiological functionality between 7–14 days.
Implantation of bioengineered mouse IAS rings.
Parallel IAS constructs in culture were implanted into C57BL/6J mice. The period of implantation lasted between 25–28 days, after which the animals were euthanized and the IAS constructs were harvested and reassessed for physiological functionality. On the day of harvesting the IAS ring, visual inspection of the implanted ring in situ revealed significant muscle attachment to the back of the mouse. Blood vessels were visible on the base of the ring. Figure 2 shows the ring still attached to the animal, with signs of neovascularization around the ring. There were no signs of inflammation or abscesses and no visual indicators of tissue involution. The growth factor pump supplies FGF only for 21 days, and the rings survived at least a week over the 21 days, without a continuous supply of FGF.
The implanted mouse IAS constructs were fixed and sectioned after physiological assessment. H&E staining showed that we were able to recover intact smooth muscle and confirm that the alignment of the smooth muscle cells was maintained in vivo postimplantation (Fig. 3, A and B). The sections were stained with Masson's trichrome to clearly identify collagen in the implanted constructs. Very little collagen deposition was observed around the outer ridge of the smooth muscle construct, as indicated by the thin blue lines in Fig. 3C. Any other collagen visualized was associated with neovascularization.
Visual vascularization was confirmed with endothelial specific antigen von Willebrand's factor (factor VIII) staining. Figure 4, A and B, shows bright green positively stained blood vessels in and around the implanted IAS ring. Moreover, the implanted smooth muscle also maintained its phenotype and stained positive for α-smooth muscle actin and smooth muscle specific h-caldesmon (Fig. 5), indicating the maintenance of the smooth muscle phenotype over 4 wk of implantation.
Western blot analysis.
Lysates from intact native mouse IAS and implanted bioengineered IAS were probed for the presence of interstitial cells of Cajal by using c-Kit and fibroblasts by using prolyl-4-hydroxylase (Fig. 6). The native intact mouse IAS displayed bands at ∼145 kDa for c-Kit and ∼61 kDa for prolyl-4-hydroxylase, whereas the bioengineered IAS did not show any bands, indicating the absence of fibroblasts or Kit-positive ICCs in the implanted construct.
Establishment of spontaneous basal tone.
Mouse IAS rings generated tone spontaneously, pre- and postimplantation when attached to the measuring arm of the force transducer with no external stimulation whatsoever. The pattern of basal tone establishment in IAS constructs pre- and postimplantation were similar (Fig. 7). The active basal tone observed in the preimplant rings was 94.7 ± 19.9 μN (n = 5) and the active basal tone in the harvested postimplant IAS rings was 114 ± 28.4 μN (n = 8). Spontaneous basal tone generation is a characteristic property of IAS physiology and it was preserved over the course of implantation.
Relaxation of basal tone by nifedipine.
Nifedipine (10 μM) resulted in a drop in the basal tone of the IAS both pre- and postimplantation as shown in Fig. 8. Nifedipine blocked basal tone by 96.5 ± 7.75% in bioengineered IAS and 85.3 ± 7.5% in postimplant harvested IAS. The bioengineered IAS as well as the postimplant IAS reached their nadirs within 100 s of treatment of nifedipine. The drop in basal tone by the dihydropyridine blocker suggests the preservation of functional L-type calcium channels in the smooth muscle, maintained during bioengineering and even postimplantation.
VIP-induced relaxation of basal tone.
Both IAS constructs pre- and postimplantation relax from their basal tone in response to 1 μM VIP (Fig. 9). The implanted IAS construct maintained the same extent of relaxation of basal tone. The harvested IAS ring relaxed 84 ± 9.7% of basal tone, whereas the preimplant counterpart relaxed 74.8 ± 19% of its basal tone. Both constructs reached their respective nadirs between 90 s (postimplant) and 120 s (bioengineered IAS) after the addition of VIP. The relaxation in response to VIP treatment indicated the preservation of VIP receptors on the smooth muscle, and the intracellular signaling mechanisms mediating VIP-induced relaxation.
Relaxation of basal tone by nitric oxide donor SNP.
Treatment of the IAS constructs with 10 μM SNP resulted in a rapid relaxation of basal tone in bioengineered IAS and postimplantation IAS as shown in Fig. 10. The average relaxation in the bioengineered IAS before implantation was 77.17 ± 1.714%. The drop in basal tone was 83 ± 10% in harvested postimplant IAS constructs. Relaxation of basal tone by SNP suggests the intact maintenance of intracellular pathways mediated by nitrergic signaling in the implanted IAS smooth muscle.
Contractility of implanted mouse IAS constructs.
IAS rings pre- and postimplantation responded in a dose-dependent manner to acetylcholine (Fig. 11) and PKC activator, phorbol dibutyrate (PdBU) (Fig. 12). The doses tested were serial from 1 nM to 1 μM, allowing up to 70 s for each individual dose to diffuse completely in the tissue bath. Both constructs responded repeatedly to increasing doses, resulting in a rapid rising contraction, which was sustained over a total of 30 min.
Figure 13 shows the response of the individual constructs to a dose of 1 μM acetylcholine, and the initial slopes of rise of contraction are almost identical between an implanted and a preimplant ring. The harvested IAS maintained the magnitude of contraction and also the sustainability of contraction for up to 300 s postcholinergic stimulation. The postimplant IAS ring maintained the membrane receptors mediating cholinergic contraction and the intracellular mechanisms involving PKC activation resulting in smooth muscle contraction.
Response to potassium chloride.
Bioengineered preimplant IAS and harvested postimplant IAS both produce rapid rising contractions in response to 30 mM potassium chloride (Fig. 14). Peak maximal KCl-induced contractions were almost equal to 1 μM acetylcholine-induced contractions. In the harvest postimplant IAS, KCl contractions were 100.2 ± 2.9% (n = 6) of acetylcholine-induced contractions. In bioengineered IAS, KCl contractions were 82.6 ± 14% (n = 4) of peak maximal agonist-induced contractions. These suggest that the integrity of the membrane's ionic characteristics as well as the mechanisms involved in electromechanical coupling in smooth muscle contraction were preserved over the course of implantation of IAS.
FI has several associated primary risk factors, with aging and sphincteric degeneration contributing in a major way to lower continence pressures (4, 7, 19). The IAS is vital in the maintenance of rectoanal continence, the restoration of which is the key to FI therapeutics. Currently available therapies like injection of bulking agents have limited results in providing a positive remedy to partially degenerated sphincters (29).
We have previously bioengineered a three-dimensional model of the IAS from a highly enriched population of sphincteric circular smooth muscle cells from rabbits and humans. It was established that physiological functionality and alignment of the constituent smooth muscle cells of bioengineered IAS rings matched closely to the IAS smooth muscle in vivo (14, 27). In the present study, IAS rings were bioengineered from a highly enriched population of circular smooth muscle cells isolated from the IAS of C57BL6 mice for the purpose of implantation.
As an extension of our previous work, we implanted bioengineered IAS rings under the skin on the backs of C57BL/6 mice. A microosmotic pump was implanted to deliver fibroblast growth factor in balanced solution to the IAS ring for 21 days. This promoted smooth muscle proliferation and angiogenesis as reported before by others (18). Moreover, the balanced solution also avoided any ischemic shocks the ring may have experienced, thus promoting a healthy neovascularization.
The animals tolerated the initial implantation surgery well, without any external signs of rejection, possibly because the cells constituting the bioengineered construct were derived from mice of the same strain. Upon harvesting the IAS ring after 28 days, neovascularization was visually confirmed by the rich pink color of the ring and the formation of blood vessels in the vicinity of the ring. Visual vascularization was also confirmed by von Willebrand's factor (factor VIII) staining (Fig. 4). Morphological assessment with routine H&E staining of cross sections of the harvested ring revealed that the smooth muscle alignment achieved during the bioengineering process was maintained during the course of the implantation (Fig. 3).
Previously, Dunn and colleagues (18) implanted tissue engineered intestinal smooth muscle constructs but failed to evoke significant contractions once these constructs were retrieved. Moreover, they seemed to have dedifferentiated into synthetic muscle upon implantation. Other work involving the injection of muscle-derived stem cells did not unequivocally address the issue of whether the cells just acted as a bulking agent or whether they were improving the inherent functionality of the smooth muscle itself (17).
Physiological functions of the IAS rings were assessed on the harvested postimplant IAS construct. These were focused on assessing the maintenance of 1) generation of spontaneous basal tone and blockade of tone by L-type calcium channel blocker nifedipine, 2) membrane receptors and intracellular signaling mechanisms for relaxation of basal tone by VIP-ergic and nitrergic stimulation, 3) contraction by cholinergic stimulation, and 4) integrity of the constituent smooth muscle cell membrane ionic characteristics in its response to potassium chloride. These properties were compared with physiological functionality of the bioengineered IAS rings before implantation.
Harvested postimplant IAS rings maintained the property of bioengineered IAS rings of exhibiting spontaneous basal tone in the absence of any external stimulation. Both sets of constructs generated similar magnitudes of active basal tone. This indicated that the implanted IAS preserved the specificity of sphincteric circular smooth muscle cell type. Basal tone was insensitive to the addition of nerve blocker tetrodotoxin. These data strongly suggested that the tone generated in our constructs was of myogenic nature, in line with previously documented work (25).
To further our physiological examination on the implanted IAS, we probed the blockade of basal tone by an L-type calcium channel blocker. It has been shown before that blocking of the L-type calcium channel using dihydropyridine blocker nifedipine lowers or blocks the generation of the spontaneous basal tone of the mouse and human IAS and the lower esophageal sphincter (9, 21). In our study, we find that 10 μM nifedipine lowers the basal tone of the IAS even postimplantation by up to 85.3 ± 7.5% (Fig. 8). Implantation thus preserves the membrane characteristics and the integrity of the major physiologically relevant L-type calcium channel responsible for the generation and maintenance of IAS basal tone.
VIP, an important component in nonadrenergic/noncholinergic relaxation of IAS and gastrointestinal smooth muscle, has been shown to act directly on receptors on smooth muscle cells causing relaxation with associated activation of PKA and PKG (6, 12, 22, 23, 26). VIP induced >90% relaxation of basal tone in the postimplant IAS as well as the bioengineered IAS rings. The extents of relaxation of basal tone were similar in harvested as well as preimplant IAS rings (Fig. 9). This indicated the preservation of the integrity of VIP-mediated relaxation of tone during implantation. The intracellular mechanisms by which VIP acts, by downstream activation of PKA, were confirmed by a direct relaxation in response to cyclic AMP in the postimplant IAS and an inhibition of VIP-induced relaxation in the presence of PKA inhibitor, H-89 (data not shown).
Nitric oxide is known to play a major role in inhibitory neurotransmission in the gut and the IAS. The rectoanal inhibitory reflex resulting in a transient relaxation of the IAS is one of the most important physiological functions of the IAS. Nitric oxide donor SNP is known to cause relaxation of basal tone in smooth muscle strips in vitro (8, 21). SNP spontaneously releases cell permeable nitric oxide in solution, resulting in a rapid relaxation. The extent of relaxation was 83 ± 10% of basal tone in postimplant constructs (Fig. 10), suggesting the preservation of NO-mediated relaxation of basal tone. The data thus suggest the preservation of both VIP- and NO-mediated relaxation of basal tone in the IAS after implantation.
The postimplant IAS ring contracts in response to acetylcholine treatment in a dose-dependent manner. Contraction of smooth muscle in response to acetylcholine is well documented and involves acetylcholine binding to the muscarinic receptor. In a G protein receptor-coupled signaling pathway, PKC is activated along with an increase in intracellular calcium concentration. Contraction pathways converge downstream to increase phosphorylation of myosin light chain to enable cross-bridge cycling and subsequent contraction (22, 28). PdBU is a membrane-permeable diacylglycerol analog that directly activates PKC bypassing the receptor-ligand interaction level, resulting in smooth muscle contraction. Our results indicate the preservation of the integrity of the membrane muscarinic receptors and the maintenance of intracellular contractile pathways; 30 mM KCl is a depolarizing concentration, leading to contraction, which is close to 100% of acetylcholine-induced peak maximal contraction, suggesting sound maintenance of basic electromechanical coupling in the implanted IAS.
Bioengineered IAS smooth muscle tissue that is already physiologically functional could be implanted to either supplement the function or replace an ailing degenerated IAS. Bioengineering is a scalable process in which the dimensions of the bioengineered ring as well as the number of cells the rings constitute could be scaled up to the species of interest. Studies have been initiated to intrinsically innervate the bioengineered tissues with enteric neurons, as a means of augmenting the physiology and bringing it closer to in vivo sphincteric function. These are important steps to be implemented before the bioengineered IAS can be used in FI therapeutics.
In conclusion, these studies provide proof of concept of a successful implantation of a three-dimensional bioengineered IAS construct that could be used to potentially treat FI associated with IAS dysfunction. Postimplant IAS rings maintain 1) the specificity of the constituent sphincteric circular smooth muscle phenotype, 2) overall IAS physiology and generation of spontaneous basal tone, 3) ionic integrity of the smooth muscle membranes, and 4) integrity of the membrane associated intracellular signal transduction pathways that mediate contraction and relaxation to physiologically relevant stimulants in a similar, if not better, manner. The implanted smooth muscle was neovascularized. The animals seemed to tolerate the implantation well for a month, without external signs of rejection. This concept could be extended toward bioengineering homologous IAS to restore inherent IAS smooth muscle contractility in FI.
This work was supported by National Institutes of Health Grants NIH/NIDDK DK071614, 1RC1DK087151 and the Training Program for Organogenesis T32HD007505.
No conflicts of interest, financial or otherwise, are declared by the author(s).
- Copyright © 2010 the American Physiological Society

subset: commoncrawl


text:
Too shy to go? Bathroom stage fright a real conditionWed Jul 6, 2011
Trump not running for presidentMon May 16, 2011
- I would guess that he never intended to run. He has to know he wouldn't get the nomination, let alone win the Presidency. If he had run and (undoubtedly) lost, he would have to deal with being a loser to Romney, Gingrich, etc. or probably worse to him, Obama. This way he can arrogantly say that he w …
Vote: Weigh in on Palin's reactionWed Jan 12, 2011
Is it OK for men to cry in public?Tue Dec 14, 2010
'Chemo curls' another kink in cancer recoveryWed Mar 23, 2011
What is 'blood libel'?Wed Jan 12, 2011
- Palin may not be directly to blame, but don't insult everyone's intelligence by saying the crosshairs are actually surveyor symbols. Palin has used firearm and hunting references throughout her "campaigning." Of course the symbols are crosshairs. Has she been surveying land in Alaska or killing cari …
'What Not to Wear' star slams Sarah PalinThu Dec 30, 2010
White House gate-crashers land TV roleTue Jun 15, 2010
In light of admitting "transgressions," what is your opinion of Tiger Woods?Wed Dec 2, 2009
Scoop: Party crashers know three rules of fameSun Nov 29, 2009
- I think they should be penalized for their actions--either with a fine or jail time. In fact, give them suspended jail time with parole, under the condition that they do not profit off this incident in any way. If they sell their story to the tabloids or TV for an interview or if they get a reality …


text:
Melhuse traded to Texas
Melhuse traded to Texas
"I'm excited about the opportunity," Melhuse said. "These things are part of the game. Hopefully, this will be an opportunity to play."
An opportunity to play was something Melhuse wasn't getting in Oakland, as he appeared in only 12 games this year and got just 26 at-bats. He was hitting .231 prior to the trade.
"I wish him well, and hopefully, he'll get an opportunity to play," manager Bob Geren said.
Melhuse obviously wasn't expecting to play ahead of starter Jason Kendall, but he was expecting to start more than six games.
"I wasn't asking to be a starting catcher, just a normal Major League backup who plays every five days or in a day game after a night game," he said. "Jason's a great player, and he's struggling a little bit this year, but it was evident [even] with his struggles there wasn't any room here for me."
Melhuse, who was in his fifth season with Oakland, will be reunited with former A's third-base coach Ron Washington, now the manager in Texas.
"It's weird, because I'm comfortable here, but playing time is tough to come by," Melhuse said. "I'm glad they gave me this opportunity to play, and I think Wash had something to do with it.
"I'll do whatever Wash wants me to do -- he's always been real good to me. I'm anxious to get over there and play my [behind] off. I'm 35, so I am not getting any younger, but I think I still have some things to offer."
The A's called up Kurt Suzuki from Triple-A Sacramento to take Melhuse's place on the roster. Suzuki was hitting .280 with three home runs with the Rivercats. He was drafted in the second round of the 2004 First-Year Player Draft out of Cal State-Fullerton.
"Hopefully, [Adam] will get an opportunity to play," A's assistant general manager David Forst said. "We'll give an opportunity for Kurt to play and get some experience."
Matt Smith is an associate reporter for MLB.com. This story was not subject to the approval of Major League Baseball or its clubs.


text:
Dice-K yields just one run in latest rehab outing
Dice-K yields just one run in latest rehab outing
BOSTON -- Daisuke Matsuzaka threw 5 1/3 innings of two-hit ball for Triple-A Pawtucket against Norfolk on Thursday night.
Matsuzaka struck out four, walked none and left the game after throwing 76 pitches (51 strikes). He surrendered a home run to the second batter he faced, Lew Ford, but that was his only blemish.
Thursday was his second start since he received a cortisone shot in his right trapezius muscle, near the shoulder and neck, on May 20. Matsuzaka is in his second 30-day rehab stint this year, his first ending on May 22.
Dice-K is recovering from Tommy John surgery and has started five games for Pawtucket. He was able to get another 30-day stint in the Minors because a different injury situation arose, which led to the cortisone shot.


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as the title infers.....im looking for both shops as were snowed under at the mo! you'll get free tea/coffee, get to work on every level of bike from a skip worthy model to a top end orange/colnago...
any questions just email me...
as the title infers.....im looking for both shops as were snowed under at the mo! you'll get free tea/coffee, get to work on every level of bike from a skip worthy model to a top end orange/colnago...
any questions just email me...
Will the remuneration package tempt me away from my high powered IT monkey job?
free tea and coffee is about all you'll get working in the bike trade!!!
I've seen a few Colnagos that belong in skips mesel'.
too far to travel but would have been up for that....
I have neither the skill nor the ability to work in your shop but I spent many happy years just up the road at RM Condor and now I'm gonna waste the rest of my day reminiscing! In fact looking at your map I think the road past your shop used to be on our Friday morning 9 mile run so I've no doubt sweated pints and pints of neat Heavy just outside your door! Oh happy days
nope the moneys not great but the bike bits are all at cost!
If bike bit's at cost then that would save me £££££££££££££££££'s
anyone? i cant believe its that hard to recruit someone to work in one of our shops....its my biggest headache.if you love bikes people and coffee its all covered.
If you were in Manchester, I'd snap your hand off!
Perhaps open a shop, near Chester and i could work there.
<grumble>Wish I could. Wife wouldn't let me leave my job and move back to Arbroath or move to Stonehaven</grumble>
nope sorry cant open nor move the shops...im gonna try the local cycling clubs..
Where are the shops ? I know a recently qualified mechanic looking for work, based in Edinburgh but he has a large motorbike...
anyone want a job in a great bike shop?
Yes, but their not hiring... BOOM BOOM
can you pass my coat over?
arbroath and stonehaven....long commute from edinburgh.
heres your coat.
if you love bikes people and coffee and don't mind being poor it's all covered
Amended for accuracy - it's a shame though that the business model doesn't enable good bike shops to pay a decent wage to bike mechanics. Plenty of people are either too busy, don't have the skills or can't be bothered to do their own spannering, yet still complain at paying a fair price for work. I suspect many of these same people don't bat an eyelid at paying £100+ per hour at a franchised garage.
The good staff aren't poor as I look after them but they're is a break even point where the wage can't go any higher...(unless they sell as many bikes as me!)
Average wage in the UK is £26K - if you're paying that I suspect you will have a queue of applicants......anyway my post wasn't intended to upset, merely to highlight the different expectations of the public when it comes to paying for skilled labour. Good luck hiring.
Are Angus Bike Chain looking for someone?
Hi Chris,
Didn't realise you were so busy when I dropped my bike off last Saturday!!
Hope this means everything gets moved over to my new frame which hopefully my in-laws are dropping off either tomorrow or Friday.
Better being busy than no-one coming through the door!!!
Alan Anderson(by the way).
If you open a branch near Reigate, Surrey, I'll work for you for free for 2 months if you train me......
We have been looking for some one for a while now too. It has been alot harder than i thought. Didn't think of putting it on here though. Good luck with the hunt.
Great..I'll just need to work out how to move both shops .....
This topic has been closed to new replies.


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Recently, Greenboro, South Carolina's mayor, Bill Knight decided city council meetings will open with prayer:
"I think this adds a very distinctly America quality and a very necessary element," he said. "We all believe in something. This is an opportunity to exercise that without infringing on the government-religion prohibition."
In the video below, Phillip Drum stands up for the rights of unbelievers and the principle of separation of church and state which should (and used to) unite all Americans and freedom loving people, regardless of creed or lack thereof. Drum is an Iraq War Veteran and president of the UNCG Atheists, Agnostics, and Skeptics. The bald man chairing the meeting is Mayor Knight.
Your Thoughts?


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text:
I need this re-worded so that it passes turn-it-in
Recovery is a process, a way of life, an attitude, and a way of approaching the day's challenges. It is not a perfectly linear process. At times our course is erratic and we falter, slide back, regroup and start again. The need is to meet the challenge of the disability and to re-establish a new and valued sense of integrity and purpose within and beyond the limits of the disability; the aspiration is to live, work, and love in a community in which one makes a significant contribution.
Recovery is an ongoing dynamic interactional process that occurs between a person's strengths, vulnerabilities, resources and the environment. It involves a personal journey of actively self-managing psychiatric disorder while reclaiming, gaining and maintaining a positive sense of self, roles and life beyond the mental health system, in spite of the challenge of psychiatric disability. Recovery involves learning to approach each day's challenges, to overcome disabilities, to live independently and to contribute to society. Recovery is supported by a foundation based on hope, belief, personal power, respect, connections, and finding self-identity. Finding one's self-identity is the most important part of recovery.
Identity is not just what you know; it is also how you know. People are not born with an identity. Rather, identity is something that evolves over time. Young children have simple identities and see things in an overly simple, generally self-serving manner. As people grow older and wiser, they identify themselves with other people, places and things in increasingly sophisticated ways and start to grow out of this initial selfishness.
People's identity is rooted in their identifications; in what they associated themselves with. What a person associates him or herself with is ultimately who that person is, for all identity is ultimately in relationship to something else. An American person identifies himself or herself as "American", for example, and that becomes part of that American person's identity.
Our identity changes often over the years from childhood through the teenage years, then we identify with our career orientation, then we go into relationships, maybe parenthood, then on through those busy years toward midlife and then the empty nest, forward to our senior years.
Mostly people identify themselves with outward expressions of themselves, careers, families, looks, clothes, home, possessions, and education. All of these things reveal some aspects of our identity to a certain extent. Often it is only when there is a crisis of change that we begin to ask if we are being true to ourselves.
Supporting personal recovery involves moving away from a focus on treating illness and towards promoting well-being. This will involve transformation, in which professional models become part of a larger understanding of the person. This understanding can be guided by the Personal Recovery Framework which is based on the four domains of recovery that emerge from accounts of people who have lived with mental illness.
Self-identity is the center of recovery. One has to personally know who they are before they can change anything about themselves.
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text:
La Mesa residents, your city officials are listening, the farmers market is here to stay and your parks are being watched.
At Thursday's all-day City Council Strategic Planning Workshop, the council heard from its city manager, fire chief, police chief, finance director, city planner, community development director, treasurer and others in discussing everything from public safety to pensions.
After ideas and information were shared at the annual workshop, City Councilman Mark Arapostathis summed up his thoughts, saying, "This is a great city; sometimes we don't trumpet that enough. We're constantly focusing on improvement, another hallmark of a great city. I'm excited about the future of La Mesa and how we are working toward the next century which shows in all the projects."
The workshop, held at its Emergency Operations Center in the new downtown fire station, offered time for reflection on the city's past endeavors while bringing forth plans for future targets of action.
Continued collaboration from the city with the La Mesa-Spring Valley and Grossmont Union High school districts in matters of safety and traffic was a hot topic. Wellness through the parks master plan was also discussed.
The city patted itself on its proverbial back for the implementation of a new pet licensing program and its work in starting to installing public Wi-Fi in city facilities to enhance connectivity for residents. Small but incremental increases in La Mesa's two key revenue sources, sales tax and property tax, are helping the city recover from the economic recession..
And while the city-run La Mesa Village Farmers' Market generated gross sales of more than $500,000 in the 2013 calendar year, which was a decline of more than 10 percent as compared to 2012, it had a net profit of nearly $12,000.
Assistant City Manager Yvonne Garrett spent time going over some of the residents' issues that were aired at two town hall meetings in February, including concerns about drugs being sold or used at Northmont Park.
Hidden camera equipment that feeds directly to the city's main police station was recently installed at Northmont, Police Chief Ed Aceves said. A citizen in the area has also volunteered to be a liaison to the department to report directly to police when something is amiss, Aceves noted. There was some talk about other parks having increased security measures added as the city's budget allows.
Parks in La Mesa were a big part of the workshop, as Garrett noted: "People care about parks in La Mesa."
Garrett spoke about the city's $75,000 grant in 2012 from SANDAG for its La Mesa Live Well Park Master Plan. Landscape architect and consultant Gary Hoyt shared ideas about fitness trails, recreational trails, urban stream restoration, security upgrades, connections to neighborhoods and more for the city's 14 parks.
City officials and elected officials shared their anticipation of partnerships that have been working on a developed Highwood Park. The park will be home to the La Mesa Arts Academy at La Mesa Middle School and the new Boys & Girls Clubs with a full gym to be built on the land, all close to the new Performing Arts Center at Helix High.
City Manager Dave Witt said the coming condominium project at the site of the old Coleman College (and before then, a bowling alley) is at least a year and a half from coming to fruition.
"They're probably somewhere close to start construction, they've finished the initial demolition," Witt said, noting that the city was braced for "new families and a spinoff for new investment in the area."
The long-planned Downtown Village Streetscape is set to begin in early summer, and Witt said the city would "work through all the special events, and work out all the logistics."
That sounded good to Councilwoman Ruth Sterling, who spoke up many times during the meeting.
"We all love La Mesa," Sterling said. "La Mesa is a pocket of goodness, of sweetness."


text:
(Reuters) – Many supporters of Barack Obama hoped his election as America's first black president might herald an era of post-racial politics, but race has been an issue his administration just can't seem to avoid.
Division and tension between black and white Americans has cropped up repeatedly over Obama's 18 months in office, hurting his popularity and distracting from his political agenda.
For the rest of this Reuters story, please go to:


text:
My Nonprofit Reviews
1177 profile views
1 reviews
Review for WAGES (Women's Action to Gain Economic Security), Oakland, CA, USA
WAGES is great, providing a valuable service to its clients and participants. i plan on using them in the future. go wages!
More Feedback
The kinds of staff and volunteers that I met were...
dignified and skilled
Ways to make it better...
i had learned about them sooner
When was your last experience with this nonprofit?
Role: General Member of the Public & i worked with them on their branding.


text:
The '90s classic Mrs. Doubtfire is getting a sequel!
Robin Williams, who played everyone's favorite disguised nanny, and director Chris Columbus are both attached to the new film, according to The Hollywood Reporter.
The sequel has reportedly been in the works for some time, but gained momentum only recently when Elf writer David Berenbaum signed on to pen follow-up to the 1993 hit.
Let's keep our fingers crossed that Miranda (Sally Field) and Stu (Pierce Brosnan) will return in the new film as cold as ever and force Daniel to adopt a new facade. Maybe as a hip cleaning lady?
Until then, we're going to jump around in celebration!
Read More:
7 Disney Channel Original Movies That Need Sequels Immediately
Kids Freak Out After Seeing A Walkman; 10 Other Things We'd Like To See Them React To
5 Things You Never Knew About 'Mean Girls'
For the latest pop culture news and voting, make sure to sign up for the People's Choice newsletter!


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Is the microfiber towel still available?
July 2 2007 at 3:48 PM
Juliet (
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Response to
Everything is still available n/m
If yes, I'd be interested and, of course, I'll pay whatever you need for international shipping.
Please let me know!
Respond to this message
Yes. Please email me :)
- AmandaPanda on Jul 2, 2007, 9:25 PM
Hi Amanda, I emailed you. /nm
- Juliet on Jul 4, 2007, 3:11 PM
Hope you got the towel by now n/m
- AmandaPanda on Jul 28, 2007, 5:06 PM
Yes, I did! It's great! Thank you Amanda! :-) nm
- Juliet on Jul 30, 2007, 5:41 AM
Woohoo! :) n/m
- AmandaPanda on Aug 5, 2007, 1:37 PM
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text:
Georgia Bulldogs Football: OL Kolton Houston Cleared by NCAA After 3-Year BattleJuly 25, 2013
Georgia offensive lineman Kolton Houston has finally been cleared to play by the NCAA after being suspended by the NCAA in 2010 for testing positive for a banned substance, according to the Associated Press (h/t USA Today).
Houston's reinstatement came on his 22nd birthday, a good bit of news for a player who has fought an arduous three-and-a-half-year battle to be able to play college football.
The whole saga started in January 2010, when Houston tested positive for the banned substance 19-norandrosterone. The problem was, Houston had a perfectly good reason for it being in his system: Doctors had prescribed him a steroid injection to help heal a shoulder injury in high school.
The NCAA rejected Georgia's appeals that the legitimate prescription of the steroid before he came to the school should override his one-year suspension. Furthermore, the substance kept showing up in his system on further drug tests in levels over the NCAA-allowed minimum, which resulted in the NCAA giving him a lifetime ban.
Unfortunately for Houston, the steroid had become trapped in his fatty tissue, meaning that he continued to test positive even though he hadn't had any subsequent injections after the one in high school that started the whole ordeal.
The University didn't give up though. According to Chip Towers of the Atlanta Journal-Constitution, the school's director of sports medicine, Ron Courson, tirelessly appealed the NCAA's decision, providing regular tests the school had conducted on Houston proving that there had been no further steroid use, despite the continued elevated levels.
The NCAA refused to provide immediate reinstatement, but agreed to reinstate him once his levels reached the NCAA-approved threshold. After trying a number of different things to reduce the levels, including surgery to remove the contaminated tissue, Houston's perseverance finally paid off.
His latest test showed his levels to be below the threshold, leading to the NCAA reinstating him on his birthday.
UGA and Kolton Houston both deserve a lot of credit for this, as they fought tirelessly to overcome a very frustrating situation. It is unknown how Houston will fit into the depth chart currently, but for now, the school is just happy to have him back on the field.
And Kolton Houston is just happy to finally have the chance to play college football.


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Vitria provides the industry's leading Operational Intelligence platform, empowering partners and customers alike to develop innovative operational intelligence solutions to analyze streaming big data, complex events, and business processes in the proper context and take real-time action. The result is faster, better decision-making.
With a rich heritage as a BPM technology pioneer, Vitria's award-winning process integration solutions provide the backbone for many Global 2000 companies' mission-critical business processes.
Vitria's technology is available to run on premise or in the cloud.
Vitria was founded in 1994 by Dr. JoMei Chang (CEO) and Dr. Dale Skeen (CTO).


text:
WKCDA commences community engagement project 'Crossover Lab Initiative' in collaboration with District Council
The West Kowloon Cultural District Authority (WKCDA) joins hand with District Council to commence the 'Crossover Lab Initiative' to showcase the distinctive arts and culture of each district using the Nursery Park in the West Kowloon Cultural District (WKCD) as the platform. The three pilot programmes will be taken forward in collaboration with Yau Tsim Mong District Council, Kwai Tsing District Council and Wan Chai District Council. Each event will be implemented with an NGO or Art Group as programme partner to encourage wide participation by the local community in arts and culture.
The three pilot programmes under this initiative, namely 'Playing Around Yau Tsim Mong Street Corners @ West Kowloon', 'Kwai Tsing Daily @ West Kowloon' and 'Experience Wan Chai @ West Kowloon' coordinated by HULU Culture, Make A Difference Institute and Hong Kong Youth Arts Foundation, will be held on 15 January, 26 February and 19 March 2017 respectively. To achieve the vision of WKCD in developing an integrated arts and cultural district, audience will be offered a wonderful experience of the multicultural Hong Kong where East-meets-West and Modernity-meets-Tradition by various activities including stage performance, handicraft workshop, bazaar, landmark art installation, family game, interactive storytelling, photo exhibition and street gourmet that will bring to WKCD the local atmosphere as well as lovers of arts, culture, handcraft and local nostalgia.
Highlighting the district's historical and cultural aspects, the first programme 'Playing Around Yau Tsim Mong Street Corners @ West Kowloon' features art installation of mah-jong tiles objects d'art and 3D paper local landmarks – Clock Tower, Yau Ma Tei Police Station and "pai dong" or stalls at Ladies' Market in Mong Kok area, and stage performance of oldies and local dancing. Upcoming next 'Kwai Tsing Daily @ West Kowloon' promotes and extends the event through two pre-event workshops 'Kwai Tsing Stories' and 'Flash Mob Theatre', and a free community paper to publish stories gathered from the district. The finale 'Experience Wan Chai @ West Kowloon' fosters the exchange between NGO or Art Group which performance, booth and workshop will be coordinated in a 1+1 partnership for martial art, juggling, craftsmanship of fire dragon, and interactive graffiti creation.
WKCDA's Chairman of the Consultation Panel Professor John Leong Chi-yan, said, "The Consultation Panel agreed to co-organise 'Crossover Lab Initiative' with District Council in early 2016 aiming to facilitate co-operation with art providers through an NGO or Art Group partnership. We wish the pre-event workshop and engagement will enhance the connectivity and the sustainability of local art and culture in a community."
WKCDA's Chief Executive Officer Mr Duncan Pescod, said, "Dedicated to become a new vibrant cultural quarter, WKCD and its Nursery Park will be the showcase of a multicultural and multifaceted Hong Kong demonstrating its distinctive characteristic through diversified art form and creation on stage."
'Crossover Lab Initiative' is a series of free events co-organised by West Kowloon Cultural District Authority and District Councils which welcome public participation. Further details are listed as below.
'Crossover Lab Initiative' Programme Schedule
Date & Time
Programme Partner
Programme Highlight
Playing Around Yau Tsim Mong Street Corners @ West Kowloon
15 Jan 2017 (Sunday)
Yau Tsim Mong District Council
HULU Culture
· Art Installation
· Traditional Handcraft Workshop
· Creative Art Booth
· Interactive Storytelling Zone
· YTM Theatre
· Local Market
· Big Stage Performance
Kwai Tsing Daily @ West Kowloon
26 Feb 2017
Kwai Tsing District Council
Make A Difference Institute
· Kwai Tsing Stories
· Flash Mob Theatre
· Master Workshop
· Sports Day
· The Fair
· Open Stage
· The Potluck
· Mini Container Terminals
· Photo Exhibition
Experience Wan Chai @ West Kowloon
19 Mar 2017
Wan Chai District Council
Hong Kong Youth Arts Foundation
· Cultural Experience (Storytelling & Craftsmanship of fire dragon)
· Stage Performance & Food Stalls
· Performing Arts (Martial Arts & Theatre Workshop)
· Photo Backdrop
· Graffiti Creation
About West Kowloon Cultural District
Located on Hong Kong's Victoria Harbour, the West Kowloon Cultural District is one of the largest cultural projects in the world. Its vision is to create a vibrant new cultural quarter for Hong Kong. With a complex of theatres, performance spaces, and M+, the West Kowloon Cultural District will produce and host world-class exhibitions, performances, and cultural events, as well as provide 23 hectares of public open space, including a two kilometre waterfront promenade.
Programme Partner:
HULU Culture (Yau Tsim Mong District)
HULU Culture is a Hong Kong non-profit organisation to promote local culture and pay keen and dedicated attention to encourage a diversified development of local culture. It believes that through the participation and sharing of people from different social strata, through a convergence of the community, and through the provision of opportunities for people to use their talents, the development of social culture in Hong Kong can be advanced.
Make A Difference Institute (Kwai Tsing District)
Founded in 2009, MaD (Make a Difference) inspires and empowers young people all over Asia to come up with creative responses to our time's challenges. It has evolved as a collaborative platform of creative changemakers that works at the intersection of creativity, entrepreneurship, innovation and discovery to bring about positive changes in Asia, and supports the growth of a vibrant creative ecology, and values cross-sector synergy and co-creation opportunities, empathetic collaboration and sustainable practices.
Hong Kong Youth Arts Foundation (Wan Chai District)
Hong Kong Youth Arts Foundation (YAF) is a charity that provides access to high quality, non-competitive free-of-charge arts experiences for all young people aged 5 to 25. Established in 1993 by Lindsey McAlister OBE, JP, YAF organises inclusive and inspirational projects that reach out to youngsters of all cultures, backgrounds, languages and abilities, and actively creates opportunities for those who are disadvantaged and underprivileged.


text:
Click Here to Buy It Now At Ray-Ban.com, $155
You can't go wrong with Ray-Bans, and the Ray-Ban Clubmaster is a timeless classic. Based off of the shape of browline glasses (a classic silhouette seen since its creation in the late 1940s), Ray-Ban sent out their Clubmaster (then called the Wayfarer Max) in the 1980s.
Clubmaster sunglasses are a nice alternative to the classic Wayfarer, while still providing the latter's clean, polished look. With a large library of color options, there's plenty of room to pick a pair that suits your taste.


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The number of men buying sex in this country has doubled in a decade to one in 10. About 80,000 women in the UK sell sex and there are an estimated 164m commercial sex transactions annually.
In the hope of reducing these figures, the government has announced zero tolerance of street prostitution and a plan to prosecute men who buy sex. Kerb crawling has been an arrestable offence since 2001, yet business is booming. A West Yorkshire police pilot scheme in 1998 and 1999 to arrest and "re-educate" kerb crawlers was shelved. An internal police report stated: "There does not appear to have been any noticeable reduction in the number of kerb crawlers since the scheme was introduced."
Sex workers reported that clients who attended "kerb-crawler rehab" continued to seek them out, but the fallout for the women was worrying. Instead of working in groups in reasonably well-lit areas and taking their punters within earshot of the other sex workers, they had to work alone in more remote areas where they were at greater risk of attack. They also had to work longer to earn the same money so, chilled and weary, they were more likely to get into a car with a man they hadn't checked out properly. Research in Sweden, where the purchase of sex was criminalised in 1999, has shown that hidden prostitution has increased, as has sex for sale on the internet. The most socially marginalised women working on the streets have suffered most, as will their UK sisters when the new zero-tolerance laws start to bite.
Asbos issued to street-based women have had a similar effect. To avoid breaching their Asbos, many have moved to unfamiliar street beats. Women who use drugs may become more chaotic away from their families and trusted drugs projects. Emma Merry was murdered after moving from her regular patch in Wolverhampton to Stoke-on-Trent because of an intense policing operation in 1994. Stoke sex workers had refused to do business with her killer as they knew he was violent, but as an outsider she did not know that and it cost her her life.
It is illegal to buy sex from anyone under 18 and it is child abuse. Adult women are often exploited too, but it is not an automatic part of the job description. At one extreme are trafficked women and those who are homeless, have expensive class-A drug habits to support, and multiple emotional problems. They need long-term support, which isn't cheap. Will these services be funded properly? At the other extreme are the high-earning women who offer escort services. They have far more control over their work and can earn more than £150 an hour. They are unlikely to argue that they are abused or that they are working against their will.
Between the two extremes are a million shades of grey: women who work in flats or saunas a couple of days a week, as they prefer it to 40 hours in a call centre; women who work on the streets intermittently for the same reason. These women have chosen to do this work, often out of economic desperation but sometimes because they like the flexible hours, their regular punters and the sex.
Violence is a problem for many sex workers and more must be done to eliminate it. Allowing women to work openly reduces the scope for attacks. A more transparent environment would also help trafficked women. The more prostitution is hidden, the greater the scope for exploitation. The proposal to allow up to three women to work together indoors rather than the current limit of one is welcome, but what of those who work in bigger establishments? Police and immigration raids are already frequent and likely to intensify.
Some police officers treat sex workers who have been raped sympathetically; others don't. All women who report rape should be treated fairly and compassionately. Many excellent projects provide sexual health and other support services to sex workers, but more are needed. Handing out free condoms to women does not reinforce their exploitation or celebrate their work. It is a pragmatic step that allows them to protect themselves from life-threatening infections. All sex workers should have access to these services.
Zero tolerance of outdoor and some indoor prostitution won't protect the most vulnerable women and won't stop them working. Instead it will drive them underground and put their lives at risk.
· Diane Taylor edits a magazine for Mainliners, a charity that works with drug users and sex workers


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Antivirus firm McAfee has hit back at critics who claim it has over-hyped the severity of Perrun, the first virus it has seen which infects Jpeg image files.
When McAfee issued its virus warning earlier this month about the W32/Perrun virus, stating that it should be treated as a serious threat to users and may mark the shape of things to come, antivirus experts at Sophos immediately countered with a press statement playing down the seriousness of the virus.
Sophos said it had the virus in its labs two days before McAfee released a warning. It had chosen not to send out an alert, according to Chris Wraight the company's technology consultant, because the virus was "really just proof of concept and a real non-event".
But McAfee's senior director, Vincent Gullotto who issued the warning, has stood by his claims over the severity of the virus.
He insists that although the virus may not be too much of a threat right now, his concerns are "more so for the future".
"I'm only saying what possibly may exist if the technology moves in that direction," added Gullotto.
Sophos' Wraight claimed McAfee has overstated the potential threat and warned that such behaviour "runs the risk of breeding complacency in users", making them less attentive during a serious outbreak.
The virus isn't yet in the wild but was sent to antivirus firms by its author and as far as both companies are aware has not infected any systems.


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A woman pedestrian was taken to hospital following a traffic incident in Haywards Heath yesterday afternoon (Wednesday March 26).
Emergency services were called to South Road just before 4pm.
A light van had collided with a pedestrian.
Traffic safety was maintained by firefighters and police as the woman was treated and then taken to hospital by ambulance.
The incident was then left with the police.


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"Listen, Howard Webb is a very good referee,'' Rodgers said. "He is well established in the Premier League and he's managed a World Cup final. I'm very confident that the game will be decided on quality and not on a refereeing decision."
Rodgers said that his Liverpool team felt hard done by when losing to Manchester United at Anfield in September. "We hope we don't have the decisions go against us that went for United,'' Rodgers said.
"In the first game a sending-off [Jonjo Shelvey] and penalty [from Robin van Persie] cost us the game. I hope they don't get the decisions that can influence the results.''
Stoke City manager Tony Pulis was warned by the FA after falling foul of the rules in April 2011 for praising the appointment of Webb for his team's FA Cup semi-final against Bolton.
In Manchester, Sir Alex Ferguson said that he hoped his side "don't suffer" any controversial decisions when facing Liverpool striker Luis Suarez.
"I think the lad is 'ladened' with controversy," he said. "I don't know whether he enjoys it or not – but it is something we hope we don't suffer from ourselves. I just hope we don't suffer from some of the decisions that have gone his way.
"Over the years the Liverpool-Man United games have been relatively free of controversy. We are the two most successful clubs in the country and you hope it's a good game."