text:
Today I saw an article on YOU Magazine's website, about the death of 'Get me to 21' organ donor campaigner Jenna Lowe. Sadly she did get an organ transplant at the end of last year but still passed away. On Facebook, below the article link many commented along the lines of "so sad", only 2 said that they are organ ... Read More »
Symptoms and treatment of Gastritis
Last week I felt decidedly short-changed, almost a month after quitting smoking. Instead of being the picture of radiant health I was more ill than I ever remember being. It started with a tummy pain between my navel and breast bone. I thought that I would get better, but I didn't. Our 10th wedding anniversary was far from romantic. I ... Read More »
Real Kids Sunglasses from OV Optics – Not too cool for school
I'll be honest, I have never given much thought to protecting my children's eyes from UV light. We have bought sunglasses for them, because glare can be a bit uncomfortable on the beach or hiking, and they look cute. Last Saturday at the SA Mom Blogs meet-up, a highlight was the presentation by Tanya (an optometrist) and Zule (a photographer) ... Read More »
Perimenopause and Pregnancy can have unnerving similarities
This post is specifically for women in the situation which I have been in recently. I spent so much time googling, trying to figure out what my body was up to, so if you have been led here by a search engine, I hope to be of help. The past 3 months have been pretty stressful. Four months later I ... Read More »
Hypochondria in a 7 year old
Just over two weeks ago, Fjord was part of a conversation about ruptured appendix. He heard that a person had his appendix removed because it ruptured. Over night he became a severe hypochondriac. Every day he talked about appendixes exploding and poison going into his body. He imagined tummy pains and was convinced that this had happened to him, more ... Read More »
Tea Tree oil fixed my sore throat
For the last few days I had a really nasty sore throat, it started with a swollen gland and progressed to real pain. I asked Cliff to look at my throat with a torch and he said that it looked like my tonsils were swollen and inflamed. I don't go to doctors unless I feel that something serious is on ... Read More »
Torn Calf Muscle – forced to lie down with a book …. woe is me
Today life seems to be saying "There are ways and means to slow you down". As Cliff was taking Fjord to school I ran to the car with his school lunch as they were pulling out of the driveway. As I ran I heard a pop and it felt like a tennis ball had hit me in the calf. I ... Read More »
Bleeding during pregnancy – a doctor almost killed my baby
Last night I read the heart wrenching post by Diane on the blog Chad Life Us about the day that she lost her 17 year old son. I cannot imagine anything worse than the death of a child, a child whom you have nurtured, cherished and watched grow over any length of time. I cannot imagine how I would ... Read More »
Mothering fail, I fed my children toxic rotten ham
This post is written for anyone who is searching the web in a panic about the results of consuming rotten, turned cold meat such as ham. After my children ate putrid ham I was told that symptoms would display within 6 – 8 hours, fortunately my children survived my flawed parenting unscathed. Incidentally the meat was used before the sell ... Read More »
Signs of concussion
I really should know not to leap out of bed, I sometimes have low blood pressure and have fainted before. Last time I keeled over backwards and cut my head open on the tiled floor resulting in 24 hours in hospital for observation. Yesterday morning I did it again, I jumped out of bed too fast at 6 am, passed ... Read More »


text:
The true method of the singing is in relaxation with nature and generally laws of health Giovanni Battista Lamperti, The Technics of Bel Canto
Giovanni and this man's father carried from the illustrious banner akin to the of voice course through the 19th one hundred year. While the method was uncomfortably short on medical analysis for a modern, industrial world (unlike your rising, competing method of the the ), it been recently long on histories relating to success spanning from that this great castrati of how the 17th century to all many successful students while teachers that emerged starting from the studios. All teachings of the two or three have accordingly ingested on a "holy writ" status among many vocalists today: either cherished and it could be ignored for their olden origins.
The Lamperti programme could perhaps remain best described once the accumulated perception and art having to do with an accomplished still , diverse and refined mtier, distilled as well as , adapted for contemporary day use. While each and every father and youngster did express specific share of reactionary disgust at popular disregard for particularly and taste in the operatic legacy, these were clearly realistic with regard to preparing their individuals for the troubles present. Unlike its father, Giovanni in fact went so a whole lot as to mention that there was naught damaging to a fabulous singer's voice within just singing the hard demands of Wagnerian and Verdian chrome. Rather, the deficiency of preliminary research and training is responsible for often the host of stressed voices that fell into casualty to the fad for bigger, higher, louder, longer safari.
Giovanni Battista Lamperti was born to 1839 in Milan. There, he the chorister at after you cathedral and canvased voice and keyboard at the sunroom. A student and later accompanist for his grand dad at the conservatory, Giovanni knew compared to anyone else the strategy his father that you learn (which he promoted descended from device castrato-teacher Bernacchi).
Appropriating it about teaching his extremely own students, Giovanni will also began teaching style at the Milan conservatory and in that case , for 20 years old in Dresden, nicely Berlin. His number one teaching arrangement am having three actually four students active at each lesson: each would get their pleasurable turn while other people observed and realized thereby. He has been said to unquestionably be a strict, exacting mentor not given so that you flattery, but the people that enthusiastically praised your students upon first rate achievement. Many of a GiovanniEURYs students came to be international opera hollywood film stars including Irene Betrothed, Marcella Cambric, Ernestine Schumann-Heink, Paul Buss, Roberto Stag no, Chris are friends . Bigamist and Franz Banach.

subset: commoncrawl


text:
Please select the appropriate category according to your ability and skiing style. Then choose the equipment you would like to book and complete the form providing all necessary data. The right size of your skis and boots will be determined when you pick them up at our rental shop. The due amount will be charged when you pick up your equipment.
If the material you have booked is not available, you can replace it with an equivalent material in the same category.
For further questions please do not hesitate to contact us at any time. Email address: email@example.com
Select the desired equipment and fill out the necessary information and click on "send booking". In short time your equipment is ready. You will pay when you pick up your skis and equipment in our rental in Selva Gardena.
Ski poles included in the price.
All prices are in euros including VAT and are valid only for online bookings.


text:
How to make buttons responsivePermalink Browser Info Environment
I was wondering what I am supposed to do to make the buttons responsive? When I shrink my browser the text disappears into the white background because the span is in it's own field and not flowing with the background block. How can I make it so the button expands in size to be able to compensate with the lines of text?
Actually the buttons are responsive and the text is flowing but it is really meant to be a 1 line text at all time because of the way it is centered vertically.
The text is centered vertically using line-height. So if the button has a height of 50px the script gives it a line-height of 50px as well so the text is centered vertically.
The result is when the button shrinks too much, the text wraps to the next line which is then 50px below the first one, so outside the button.
You should choose a button size that accommodates the space available.
With an icon at the left or right of the text or no icon, the text wraps normally following the behavior I described above


text:
Challah Baking Class with Shainy
Next Class: Thursday, September 26, 2019 - 7:00 pm
Special Round Challah for the High Holidays.
Instruction will be hands on. You will get a chance to make Challah dough from scratch
and learn to braid a traditional Challah. You'll make your own Challah that you can take home and impress your friends and family with! We will also discuss the significance of Challah and explore the beauty and Kabbalah of the Challah baking tradition.
Classes are small and by reservation only!
To reserve a space for the class – please email: email@example.com
*Please note: There is a suggested donation of $10 to help cover costs. Thank you!
Reserve Early! Classes fill up quickly!


text: For You How to Talk With Clients About Hair Loss 10.30.2015 Hair loss is very common but nobody talks about it. Break the silence and give your client hair loss solutions on how to make hair appear more voluminous. By Erin Gignac Hair loss is very common but nobody talks about it. Despite the current narrative of our society, your job as a stylist is to try and raise the issue to best help your loyal clients. While some are open about addressing it, others may be afraid to speak up. Approach the topic with a sensitive question once they settle into your chair: "Do you have any concerns with your hair?" If your client confides in you, teach them about hair loss and what options they have moving forward. For starters, they're not alone! In fact, hair loss is just as common in women as in men—the patterns of balding on the scalp just appear differently. The wide-held view that hair loss is a token trait of aging is also misleading. Almost two-thirds of men experience hair loss by the age of 35, with the main cause of hair loss being heredity. Yes, the good old gene pool may be causing your clients' agony. They can thank their mom and dad for this one. Genetics are the main cause for men and 20 to 30 percent of women, according to Modern Salon. This trait can be inherited from the mother or father's side of the family. Don't let your clients assume that their cause is genetic, though. Suggest that they see a dermatologist for a scalp biopsy. (Find a doctor in the area using this "Find a Dermatologist" page: aad.org/find-a-derm.) Other hair loss in women can be caused by pregnancy, weight loss or other hormone changes and imbalances, which may just require some time to take their course. However, if the hair loss may be caused by medication, diet or stress, suggest they find a solution by consulting a doctor. Other causes for this problem include: (1) the immune system attacking the hair follicles, known as alopecia areata; (2) scalp infections; (3) skin disorders like lupus; (4) radiation therapy; and (5) tight hairstyles like cornrows and pigtails. Instill hope in your clients by ensuring that things can be done to help with their hair loss. The first place for them to start is the scalp. Many products on the market clean the scalp and provide vital nutrients to the area to promote growth. Remind clients that they need to make an ongoing commitment when they use hair loss products. Usually they won't see results until a month of treatment. If they stop using the treatment, they will lose the benefits and end up where they started. In addition to offering hair loss products and encouragement, you can suggest that they improve their diet, upgrading to leafy greens, beans, tofu or red meat to get more iron, biotin and zinc into their diet. Scalp massages can also increase circulation and stimulate the hair follicles. There are also some tricks you can apply while they're in the styling chair. Here are some fast ways to make hair appear more voluminous: • Switch their part to create fast volume. • Air-dry or blow-dry the hair on the cool setting and use gentle boar bristle or nylon brushes on the hair to prevent breakage. • Suggest they try a shorter haircut for more volume. • Color their hair. This will expand the hair shaft and make strands appear thicker. • Highlight their hair to match the lightness of the scalp, disguising thinning hair. • Sprinkle hair fibers onto their scalp for instant results. • Teach them how to use certain products that will provide thickness and volume to their hair. There is also another alternative: tell them to embrace the thinning hair! There are always other ways to express self-confidence such as clothing, accessories and fitness routines. This method also saves time! Instead of worrying about hair loss, they can spend quality time making self-improvements that will matter a lot more than how much hair they have. Erin Gignac covers stories for Paramount Beauty. Follow her on Twitter and view more of her work here. Share This Tweet


text:
Justrite Manufacturing
Justrite 30 Gallon Corrosive Sure Grip EX Safety Cabinet - 893002
- Cabinets : JR-893002
- Shipping:
- Calculated at Checkout
- Justrite
- Gallon Capacity:
- 30 Gallon
- Door Style:
- Manual Close
- Cabinet Category:
- Acid/Corrosive Storage
Justrite Sure Grip EX Classic Safety Cabinets for Corrosives
Justrite's 893002 blue steel safety cabinets for corrosives have the same quality safety features as the cabinets for flammables. Al models come with double walled construction, dual vents, grounding wire connections, adjustable shelves, leakproof sills, three point self latching doors and leveling feet. Polyethylene liner for the bottom sump helps resist aggressive chemicals. Easily remove the liner for quick cleaning of drips and leaks. An all epoxy baked on powder coat finish, inside and out, provides increased chemical resistance.
Manual close models meet OSHA and NFPA 30. Cabinets are FM approved and come with a ten year warranty.
Justrite 893002 Corrosive Safety Cabinet Specifications:
Capacity: 30 Gallons
Number of Doors: 2
Door Type: Manual Close
Number of Shelves: 1
Adjustable Shelves: Yes
Exterior Dimensions: 43" W x 44" H x 18" D
Interior Dimensions: 39.5" W x 39.625" H x 14.562" D
Approvals: FM
Compliance: NFPA, OSHA
Weight: 239 lbs.
Please note: Image shown may not reflect optional door specifications
Click Here for other Justrite Manufacturing Products
Click Here for other options to the 893002 Justrite Manufacturing 30 Gallon Corrosive Storage Cabinet


text:
COVID-19 cases have now surpassed 100,000 worldwide since the flu-like coronavirus broke out from Wuhan in China.
The total number of cases now stands at 100,055 as of 8 a.m. ET on Friday, according to data compiled by John Hopkins.
The majority of the cases are in mainland China, followed by South Korea, Iran and Italy. In the United States, there are at least 233 cases and 14 deaths, according to John Hopkins.
At least 3,398 people have died due to the virus globally.
On Thursday, the World Health Organization called on all nations to "pull out all the stops" to fight the COVID-19 outbreak as it continues to spread Worldwide.
"This epidemic can be pushed back but only with a coordinated and comprehensive approach that engages the entire machinery of government," WHO Director-General Tedros Adhanom Ghebreyesus said during a press briefing at the agency's headquarters in Geneva.
"We're calling on every country to act with speed, scale and clear-minded determination."


text: A lawyer, a host, and a comedian mix a cocktail that will make you hear real opinions. This episode they discuss whether Kavanaugh's sexual assault allegation should be investigated before confirmation hearings proceed; Julie Chen's departure from The Talk due to sexual assault allegations against her husband Les Moonves; and the 70th Primetime Emmys. If you consider yourself a conversational mixologist, then hit play, until the bartender cuts you off.


text:
The location of Helicobacter pylori in the gastric mucosa of mammals is defined by natural pH gradients within the gastric mucus, which are more alkaline proximal to the mucosal epithelial cells and more acidic toward the lumen. We have used a microscope slide-based pH gradient assay and video data collection system to document pH-tactic behavior. In response to hydrochloric acid (HCl), H. pylori changes its swimming pattern from straight-line random swimming to arcing or circular patterns that move the motile population away from the strong acid. Bacteria in more-alkaline regions did not swim toward the acid, suggesting the pH taxis is a form of negative chemotaxis. To identify the chemoreceptor(s) responsible for the transduction of pH-tactic signals, a vector-free allelic replacement strategy was used to construct mutations in each of the four annotated chemoreceptor genes (tlpA, tlpB, tlpC, and tlpD) in H. pylori strain SS1 and a motile variant of strain KE26695. All deletion mutants were motile and displayed normal chemotaxis in brucella soft agar, but only tlpB mutants were defective for pH taxis. tlpD mutants exhibited more tumbling and arcing swimming, while tlpC mutants were hypermotile and responsive to acid. While tlpA, tlpC, and tlpD mutants colonized mice to near wild-type levels, tlpB mutants were defective for colonization of highly permissive C57BL/6 interleukin-12 (IL-12) (p40−/−)-deficient mice. Complementation of the tlpB mutant (tlpB expressed from the rdxA locus) restored pH taxis and infectivity for mice. pH taxis, like motility and urease activity, is essential for colonization and persistence in the gastric mucosa, and thus TlpB function might represent a novel target in the development of therapeutics that blind tactic behavior.
Helicobacter pylori is a highly motile, urease-positive, gram-negative pathogen that establishes lifelong infections of the gastric mucosa of much of the world's population (8, 29, 35). These bacteria live deep in the mucus layer, near the underlying epithelial cells and away from the highly acidic lumen. H. pylori must also survive direct exposure to stomach acid during the initial colonization process or perhaps during repopulation following failed eradication therapy. Mutational studies have established both motility and functional urease activity as essential for colonization in animal models of infection (4, 14, 26, 38). Infections with H. pylori tend to be antral predominant, but little is known regarding how bacteria ultimately select the site for colonization. It has been suggested that gastric acidity dictates the site of colonization (1, 17, 34), and in areas of endemicity, constant ingestion of bacteria, particularly by young children, exposes all areas of the stomach to colonization, allowing infections to predominate in the more permissive regions such as the less acidic antrum.
Most H. pylori bacteria are located deep within the gastric mucus layer, being distributed within ∼30 μm of the mucosal epithelial cells, and on histology can be observed within the gastric crypts (13, 25). Persistence in the gastric mucus must depend on tactic behavior and motility to avoid being eliminated by the constant turnover of gastric mucus. Direct studies by Schreiber et al. in H. pylori-infected Mongolian gerbils demonstrated that bacteria oriented within pH gradients, and in the absence of these gradients, bacteria were found throughout the gastric mucus (25). Their studies also showed that inversion of bicarbonate/CO2 or urea/ammonia gradients did not alter orientation of bacteria within the pH gradient, suggesting that pH taxis may be dominant over other chemotactic responses. These observations seem to suggest that H. pylori can sense and respond to temporal changes in local acidity by coordinating tactic signal transduction to the flagellar motors to remain in or seek out an optimal pH zone. Remarkably, the chemoreceptors and signal transduction networks associated with pH taxis have not been identified.
In Escherichia coli, pH-tactic responses to permeant weak acids and bases are modulated by chemoreceptors Tsr and Tar, respectively, which monitor changes in cytoplasmic pH (15, 32). In contrast, the cytoplasmic pH of H. pylori does not vary more than 1 pH unit in response to strong acids over a pH range from 1 to 6 when urea is present (28). Under these conditions, the pH-gated urease activity contributes to buffering of the cytoplasm, pH stasis, and acid survival (26, 28, 38). It is conceivable that behavioral responses to strong acids differ from weak-acid responses or even involve energy-tactic mechanisms that respond to the ΔpH component of the proton motive force. Energy taxis is best exampled by behavioral responses of bacteria to molecular oxygen (aerotaxis), for which aerotaxis receptor Aer transduces chemotactic signals based on the redox status of a bound flavin (19, 24). While Campylobacter jejuni, a close relative of H. pylori, contains an aerotactic chemoreceptor system (11), these genes are not present in the Helicobacter sequenced genomes (2, 31). The H. pylori genome encodes three canonical membrane-spanning chemoreceptor proteins (TlpA, TlpB, and TlpC) and one soluble chemoreceptor protein (TlpD) (3, 5, 9, 23, 30). While several groups have demonstrated chemotactic responses to a variety of substrates including urea, bicarbonate, and amino acids (3, 5, 22, 40), the substrate specificities of these chemoreceptors have not been determined.
In the present study, we have developed an assay system to track and measure pH-tactic behavior of H. pylori, and through systematic mutational analysis of each of the chemoreceptor genes, we identified tlpB as necessary for pH-tactic responses. Using a highly permissive interleukin-12 (IL-12)-deficient mouse infection model (13), we show that only tlpB mutants are defective for colonization (restored by allelic complementation in the rdxA locus), suggesting that pH taxis, like urease activity and motility, is essential for colonization and persistence in the gastric mucosa.
Bacterial strains and growth conditions. H. pylori strains SS1 and motile KE26695 (KE88-3887) were grown under humid microaerobic conditions at 37°C on brucella-based medium supplemented with 7.5% newborn calf serum (Sigma) and antimicrobials as previously described (10, 18, 21). Escherichia coli strains were cultured in Luria-Bertani medium and supplemented as required with appropriate antibiotics.
Generation of tlp allelic-replacement mutants.Figure 1 depicts the genetic organization of the four tlp genes of H. pylori KE26695, and primer sets used to generate mutants are presented in Table 1. The vector-free allelic-replacement mutagenesis strategy used to generate tlp-deficient mutants is outlined in Fig. 2 (6). Either the kanamycin resistance (Kmr) cassette aphA3 from pDH37 (10) or chloramphenicol resistance (Cmr) cassette cat originating from Campylobacter coli (37) was amplified with the appropriate primers listed in Table 1. H. pylori genomic DNA was isolated as previously described (13). Two pairs of gene-specific primers designated P1/P2 and P3/P4 for each tlp gene were used to PCR amplify the up- and downstream regions, respectively, of each target gene in order to produce fragments of ∼500 base pairs (bp) flanking the region to be deleted. The P2 and P3 primers contained 5′ leaders complementary to the E1 and E2 primers, respectively, for the appropriate antibiotic cassette and 20 to 21 bp of gene-specific sequence. The standard PCR in 50 μl contained 5 μl 10× PCR buffer, 1 μl of SS1 DNA (250 to 500 ng) or plasmid containing antibiotic cassette (1 to 2 ng), 200 μM (each) deoxynucleoside triphosphates, 300 nM of each primer, and 0.75 μl (2.5 U) of high-fidelity DNA polymerase (Roche Diagnostics). The PCR conditions were as follows: 5 min at 94°C, followed by 35 cycles of 30 s at 94°C, 30 s at 54°C, and 1 min at 72°C, and a final extension of 7 min at 72°C. The amplicons were gel purified with a QIAquick gel extraction kit (QIAGEN), and the flanking amplicons and the drug resistance cassette amplicon were amplified in a second PCR with P1 and P4 primers for each tlp gene as originally described by Chalker et al. (6), with a modification of four precycles at 54°C. The resulting long amplicons were either directly transformed into H. pylori (36) or cloned into the pBC(ks) vector (Stratagene). The vector-free protocol was used to directly construct tlpA (HP0099), tlpB (HP0103), and tlpD (HP0599) gene replacements of SS1 and KE88-3887. The tlpC (HP0082) construct generated by the vector-free method was first cloned into plasmid pBC(ks) by blunt-end ligation at the EcoRV site, after filling in the A overhang of the amplicon. After transformation of E. coli DH5α, plasmid DNA was isolated and used to transform SS1 and KE88-3887. All tlp knockout constructs were verified by diagnostic PCR techniques to confirm loss of the tlp gene and replacement with the appropriate cassette. The diagnostic PCRs comprised sets of DNA primers designed to hybridize within the aphA3 or cat cassette paired with primers hybridizing to distal chromosomal sequences, and to confirm deletion of the respective tlp gene, primers were designed to internal sequences of the respective gene. All amplicons from validations of gene replacements were cloned into pBC or pBS vectors and sequenced in our lab (Center for Functional Microbial Genomics and Host Defense core facility) on a Beckman CEQ8000 automated DNA sequencer (Beckman Coulter).
Construction of pRDX-C vector and complementation of the ΔtlpB mutant.The pRDX-C vector was constructed in pBlueScript KS(+) as essentially described by Smeets et al. for cloning rdxA sequences into pBC (27). The 5′ and 3′ portions of rdxA were PCR amplified from H. pylori KE26695 chromosomal DNA with the primers listed in Table 1. Amplicons were sequentially cloned into pBlueScript following restriction digestion (SacI and XbaI for the 5′ rdxA and XhoI and KpnI for the 3′ rdxA), creating pRDX. The cat cassette was PCR amplified using the primers listed in Table 1 and, following treatment with T4 polymerase, the blunt-ended DNA was cloned into the EcoRV site of pRDX, yielding pRDX-C. The forward orientation (same orientation as rdxA) of the cat cassette was confirmed by restriction analysis and DNA sequencing.
Wild-type (WT) tlpB, with its flanking regions, was PCR amplified from H. pylori SS1 chromosomal DNA with primers TLPBCompF and TLPBCompR listed in Table 1. The ∼2.1-kb fragment was digested with BamHI and EcoRI and cloned into similarly restricted pRDX-C, yielding pRDX-C::tlpB (see Fig. 6A for genetic organization). Following transformation into E. coli DH5α (ampicillin resistance/Cmr selection), the plasmid was naturally transformed into SS1 ΔtlpB and plated on brucella agar supplemented with 20 μg/ml each of Km and Cm (no selection on metronidazole). The resulting SS1 ΔtlpB/tlpB+ strain was screened for correct chromosomal insertion of tlpB in the rdxA locus by PCR.
pH taxis assay.pH-tactic assays in 50- or 100-μl capillary tubes were modeled on prior work with enteric bacteria and H. pylori (9, 39). Capillary tubes were filled with suspensions of H. pylori strains (SS1 or KE88-3887) in phosphate-buffered saline, pH 7.0, containing 1 mM MgCl2 or in brucella broth. The ends, one of which contained HCl (pH 0.1 to 2), were plugged with 2% agarose, and the tactic behavior was visualized under low power with a Zeiss light microscope (160× to 400×). Strains of E. coli and Proteus mirabilis (defective for pH and negative chemotaxis) (39) served as controls. A wet-mount microscope slide assay was developed in which a drop of bacterial suspension (optical density at 660 nm [OD660] of ∼0.1) was covered by a no. 1 coverslip (22 by 50 mm) that was sealed around the perimeter (three sides) with high-vacuum grease (Dow Corning). The slide was visualized with the 40× objective of an inverted phase-contrast microscope (Olympus IX71) equipped with a video capture system (Evolution QEi; Media Cybernetics) and image analysis software package (ImagePro Plus; Media Cybernetics) that enabled continuous monitoring in still or video modes of data collection. The software allowed tracking of individual bacteria, permitting time-based measures of tactic behavior. Once baseline bacterial motility had been recorded, a drop of diluted HCl (pH 0.1 to 3.0 or medium control) or HCl supplemented with 5 mM urea was placed along the open side of the coverslip and, following a few seconds to enable slight mixing to cease, tactic behavior was recorded. All recordings were performed in triplicate to ensure reproducibility and all studies were performed with different batches of bacteria grown to a common OD, to ensure reproducibility of measurements. Individual bacteria were tracked (five each) to obtain a composite of swimming and arcing behavior, since arcing behavior correlated with tactic responses.
Chemotaxis and aerotaxis assays.Motility of all strains was examined microscopically by wet-mount assay. Nonmotile KE26695 (TIGR strain) served as a visual control. General chemotactic behavior (qualitative) was assessed in semisolid medium prepared with brucella broth containing 0.3 to 0.35% agar in standard petri plates. The plates were inoculated with 1 μl of matched bacterial suspensions (OD600 of 0.1) by stabbing the plastic tip of a pipette into the soft agar. The plates were then incubated for 5 days at 37°C under microaerobic conditions. The outward migration of chemotactic bacteria was measured (diameter) every 24 h. Aerotaxis was observed in semisolid brucella broth (5 ml in 13-ml screw-cap test tubes) by inoculating the medium below the surface with 100 μl of 0.1-OD suspension and then visually tracking the migration of an aerotactic band as it migrated up to the surface of the medium. As a control for these experiments, aerotactic Campylobacter jejuni strain H840 was used as previously described (16).
Mouse infection protocols.IL-12-deficient mice (C57BL/6 p40−/−) were bred in-house at the Carleton Animal Care Facility, Dalhousie University, Halifax, Nova Scotia, Canada (13). C57BL/6J mice (Jackson Laboratories, Bar Harbor, ME) were maintained in the University of Virginia School of Medicine animal quarters. Animals were bred under isolator conditions and placed in conventional housing for at least 4 weeks prior to experimentation. All animals were given access to water and commercial chow throughout the course of the experiments. Animals were sacrificed using CO2 asphyxiation. Animal protocols were approved by the animal ethics committee of the Dalhousie Medical School animal care facility and the University of Virginia.
To ensure that mouse-colonizing H. pylori strain SS1 was fully infectious, the strain was passaged two times through mice (3 weeks each), and then all four tlp mutations were created newly in this strain by the allelic replacement methods described earlier. Tactic phenotypes for the newly constructed mutants were verified prior to mouse infections. Mice (groups of five) were infected with 0.2 ml of tlpB, tlpC, and tlpD mutants, WT SS1, 0.2 ml saline (mock infection), and a non-mouse-colonizing strain G27 by oral gavage (three times over a 5-day period). The SS1 ΔtlpB/tlpB + complement and wild-type SS1 strains were used to infect C57BL/6J mice as described above, except that two infections over 3 days were used. All animals were sacrificed at 3 weeks postinfection, and sections of stomach tissue were weighed, homogenized, diluted, and plated on Columbia blood agar plates supplemented with Dent's antibiotics for microbiological counts (13, 18). Infectivity is reported as the number of animals colonized of the total infected, and the microbial load in the stomach was determined from the means and standard deviations of the counts of CFU/g stomach material (triplicate plating) from the infected animals of each group.
Phylogenetic and protein structure analyses.Motif searches utilized SMART software (http://smart.embl-heidelberg.de ), homology searches used a BLAST search of microbial genomes (http://www.ncbi.nlm.nih.gov/sutils/genom_table.cgi ), and alignments of closely related sequences were done with CLUSTALW (http://www.ebi.ac.uk/clustalw/ ). Protein-protein interactions were evaluated using the Hybrigenics PimRider, and gene organizations as protein structure analyses were examined on the Pedant web site (http://pedant.gsf.de/ ).
Development of pH taxis assays.We initially set up pH taxis assays in capillary tubes, modeled on prior work with enteric bacteria (39). In these assays, E. coli strains formed a chemotactic band within a few millimeters of the HCl-agar-plugged end, and this band migrated away from the acid. In contrast, Proteus mirabilis, which is defective for negative chemotaxis (39), did not respond to the acid gradient and quickly became nonmotile as the acid diffused down the capillary tube. However, due to light distortion with round capillary tubes, we were unable to utilize the more quantitative video analysis system to track tactic responses of control bacteria or H. pylori. We next investigated the possibility of using a simple wet-mount assay on a standard microscope slide to visualize pH-tactic behavior. The advantage of this assay is that it provides a much wider field of view to fully assess population behavior. For this assay, bacterial suspensions of H. pylori in brucella broth were seeded onto the microscope slide and covered with a coverslip sealed on three sides with vacuum grease. Brucella broth cultures were used directly, as collection of bacteria by centrifugation and resuspension resulted in a population of poorly motile bacteria as previously noted (9).
Prior to acid exposure, baseline motility was video recorded. Most of the bacteria were motile but clearly sluggish, and swimming patterns were essentially random (Fig. 3A) (see Video S1 [http://intmed.med.virginia.edu/hoffmanlab/ ]). In response to acid, bacterial motility increased, and the majority of bacteria could be observed swimming in large clockwise arcs that were particularly sharp in the most concentrated area of acid, such that the net population movement was away from the acid (Fig. 3B) (see Video S2 [http://intmed.med.virginia.edu/hoffmanlab/ ]). Arcing or swimming in wide circles has been noted by others for H. pylori (9, 23). The arcing behavior differed from the typical straight-line swimming of enteric bacteria. Flagella are bundled (one to five) at one pole, but it is not known how rotational activity promotes arcing. Tumbling behavior was not appreciably observed with H. pylori; rather, the bacteria would stop and move a short distance in a straight line, then resume arcing behavior, but there was no reversal of direction of the arc. The net result of this arcing behavior was a collective movement of the bacterial population away from the strong acid, and within minutes, there was an absence of motile bacteria within the now acidic field of view (see Video S2). As the bacteria moved away from the acid and, presumably, to a more neutral region, bacteria concentrated into a dense band (wall) of organisms, and motility became sluggish to nonexistent as depicted in Fig. 3C (see Video S3 [http://intmed.med.virginia.edu/hoffmanlab/ ]). In response to solutions of higher pH (>3 or medium alone), no tactic behavior was noted, eliminating fluid streaming as a cause of the tactic behavior. To test whether the wall of bacteria was nonviable, we added another drop of 0.1 N HCl, and in response to the additional acid, the bacteria within the wall became highly motile once again and engaged in another round of frantic arcing taxis away from the acid source and formed a new wall some distance from the previous one. This pH-tactic response was highly reproducible and was observed with a variety of H. pylori strains in our collection. In contrast to enteric bacteria (39), H. pylori did not swim toward the acid (moving up the pH gradient to a slightly acidic optimal pH zone), so the tactic response observed appears more consistent with negative chemotactic (avoidance) behavior. The addition of 5 mM urea to the 0.1 M HCl did not alter the pH-tactic behavior of H. pylori but did protect those bacteria directly exposed to the highest concentration of acid (ordinarily rendered nonmotile in HCl alone) (data not presented).
Using a software feature, we tracked the movement of individual bacteria in the presence or absence of acid. In the absence of acid (Fig. 4A), bacteria exhibited sluggish straight-line swimming (linear velocity of 11.4 ± 1.7 μm per s) that was essentially random (Table 2). In response to acid, bacterial motility increased dramatically, and individual bacteria swam in arcs whose net activity was away from the acid source, as depicted in Fig. 4B. Table 2 provides data on motility, tactic behavior, and arcing behavior (based on tracking five organisms per sample). Note that the relative increase in linear velocity is influenced by the degree of arcing.
To determine whether H. pylori also displayed aerotactic behavior, we examined semisolid medium in a test tube for appearance of an aerotactic band as has been described with other bacteria (19). We could not demonstrate aerotaxis with motile wild-type strains of H. pylori in soft agar (data not presented). Aerotactic behavior was observed with C. jejuni strain H840 under identical conditions. Microscopic examination of wet mounts can often reveal aerotactic behavior, especially around small air bubbles (commonly observed with Campylobacter strains), and in the case of H. pylori strains, we never observed such behavior. Therefore, in the absence of clear evidence for aerotactic behavior by H. pylori, aerotaxis was not further pursued in this study.
Isolation of pH taxis mutants.In other bacteria displaying tactic responses to changes in pH or oxygen (aerotaxis), methyl-accepting chemoreceptors (MCPs) with similarities to Tsr or Aer proteins are often involved (19, 24). Examination of the two annotated genomes of H. pylori revealed four orthologs of MCPs, tlpA, tlpB, tlpC, and tlpD (2, 31), none of which share any appreciable homology in the N-terminal regions with similar sequences of Tsr or Aer. Furthermore, none of the Tlp proteins shares any homology with aerotactic proteins (CetA and CetB) of Campylobacter jejuni that were previously shown to mediate aerotaxis (11). To analyze the roles of MCP genes in pH taxis, knockout mutations were created in each of the four tlp genes. The advantage of this high-throughput vector-free technology is that amplicons go unaltered directly into H. pylori by natural transformation (6). PCR-based analysis was performed on each of the mutants, and since the replaced cassette (aphA3 or cat) is substantially smaller than the tlp genes, mutant constructs were readily distinguishable from the respective intact genes based on the size of the amplicons generated. Based on the observation that none of the clones picked contained doublets (mutant and WT amplicons), together with additional PCR-based validations, we concluded that all tlp knockouts were authentic.
Assessment of motility and chemotaxis.All tlp knockout mutants of SS1 and motile KE26695 were motile, as visualized by wet mounts, and were video recorded (data not presented). Since previous studies had shown that tlpA, tlpB, and tlpC mutants were chemotactic as measured in brucella-based soft agar (3), we examined all four tlp knockout mutants for similar tactic behavior by this assay. As seen in Fig. 5A, all mutants (the tlpA mutant is not depicted) exhibited chemotactic behavior, as shown by their ability to migrate outward from the central inoculum. The results presented are for the SS1 strain and mutants. Note that the tlpB and tlpD mutants seemed to be slightly less chemotactic than the WT and a tlpC mutant as measured at 5 days.
Defects in pH taxis.Each of the tlp mutants was screened for pH-tactic responses by the wet-mount slide assay and was visualized by phase contrast microscopy and video recorded. Our studies established that tlpA, tlpC, and tlpD mutants displayed pH-tactic behavior, though there were some noted differences compared with WT SS1. Video tracking of motility in the presence or absence of HCl showed that tlpC mutants were hypermotile and tended to swim in straight lines regardless of acidity. The linear velocity of this mutant was ∼57 μm per second (Table 2), and these bacteria rapidly formed a wall of less- motile bacteria. In contrast, tlpD mutants tended to swim in arcs in the absence of acid and to tumble more frequently. In response to acid, tlpD mutants were less robust in negative chemotaxis and wall formation. The linear velocity was decreased by nearly half by exposure to acid (Table 2). Motility measurements were not collected for the tlpA mutant. In contrast to the pH-tactic behavior of the other mutants, tlpB mutants, while displaying increased motility in response to acid (Table 2), showed no tactic response (Fig. 3D) (see Video S4 [http://intmed.med.virginia.edu/hoffmanlab/ ]). The behavior of tlpB mutants was similar to that observed with P. mirabilis, a bacterium that does not display pH taxis or negative chemotaxis (39). As the acid moved across the slide, large numbers of bacteria became nonmotile, and the typical clearing of the field observed with pH-tactic strains was not observed with the tlpB mutant. The noted random motility also included some arcing and tumbling behavior, but no taxis. If 5 mM urea is added to the HCl, bacterial motility is not inhibited by acid, presumably due to the protective action of urea hydrolysis. These results suggest that TlpB function is required for pH taxis of H. pylori.
Allelic complementation.To determine whether TlpB function was necessary for pH taxis, the SS1 ΔtlpB mutant was complemented by cloning a wild-type tlpB gene into the rdxA locus. This was accomplished by first cloning the tlpB gene into the multiple cloning sequence region flanked by rdxA sequences as shown in Fig. 6A and was validated by PCR as demonstrated in Fig. 6B (note two amplicons in SS1 ΔtlpB/tlpB + complement representing the mutant allele and complement allele). Expression of tlpB under its own promoter in the rdxA locus of SS1 ΔtlpB/tlpB + restored pH-tactic behavior, as demonstrated by migration of the bacteria away from the acid and the accumulation of bacteria into a wall as observed with wild-type SS1. The bacteria regained the arcing motility in response to strong acid and displayed pH-tactic behavior as described for WT SS1 (data not presented). Moreover, as shown in Fig. 5B, the ΔtlpB complement regained full chemotaxis in the soft agar assay, suggesting that in addition to defects in pH taxis, the tlpB mutation also affects other chemotactic behaviors or possibly full motility.
The tlpB mutants do not colonize mice.Based on the work of Schreiber et al. (25), one would predict that defects in pH taxis should manifest as deficiencies in mouse colonization. To test this possibility, we infected highly permissive IL-12 (p40−/−)-deficient mice with the tlpB, tlpC, and tlpD mutants and determined the colonization efficiency and CFU/g of stomach material for each set of mice at 3 weeks postinfection (Table 3). We had tested tlpA mutants in an earlier mouse experiment and obtained results similar to those reported by Andermann et al. (3). As seen in Table 3, bacterial densities for tlpC and tlpD mutants were comparable to those obtained with the WT SS1 strain (∼107 CFU/g). In contrast, tlpB mutants did not colonize IL-12-deficient mice (two of five animals), and bacterial counts for tlpB mutants were >5 log units lower than for the other tlp mutants, similar to the colonization efficiency determined for strain G27 (<100 CFU/g), which does not colonize mice (3, 7). To determine whether restoration of pH-tactic behavior to the ΔtlpB mutant could also restore mouse infectivity, we infected four C57BL/6J mice with the ΔtlpB/tlpB + complement and compared infectivity to WT SS1. As seen in Table 3, complementation of the ΔtlpB mutant restored infectivity to within 1 log unit of that obtained with WT SS1 (4.6 × 104 for the complement versus 3.3 × 105 for SS1). It should be noted that the colonization density of SS1 strains and mutants for C57BL/6J mice is ∼2 log units lower than for the IL-12-deficient mouse (13). These studies indicate that pH-tactic behavior mediated by TlpB is essential for mouse colonization by the SS1 strain.
Protein analysis.The TlpB protein contains two membrane-spanning domains in the N-terminal half of the protein (amino acids 9 to 32 and 211 to 231) and methyl-accepting chemotaxis domains in the C-terminal cytoplasmic half. A HAMP domain (a domain commonly found in histidine kinases, adenylyl cyclases, MCPs, and phosphatases) is located in the cytoplasmic domain between amino acids 213 and 284. However, the HAMP domain does not include a PAS (signaling) domain which in aerotaxis protein Aer contains bound flavin (19). The HAMP domain does not share amino acid similarity with the HAMP domains of the Tsr protein of E. coli, but the locations of basic amino acids and acidic amino acids seem to conserve the charge distribution that is required for cytoplasmic pH sensing in E. coli (19, 32) (Fig. 7). BLAST searches show that the N-terminal 250 amino acids are unique to H. pylori strains J99 and KE26695. There is homology in this region with a Helicobacter mustelae tlp mutant (35% identity and 54% similarity), which also colonizes the stomachs of ferrets. There was no similarity with proteins from Helicobacter hepaticus or with other species of epsilon proteobacteria (Campylobacter spp. and Wolinella succinogenes). Moreover, BLAST searches of the entire protein database identified no significant homology to any other protein that might provide clues as to how TlpB senses changes in pH. The 250-amino-acid region predicted to be exposed to the periplasm contained no obvious motifs detected by SMART or similar protein analysis software. Protein interaction information on HP0103 on the Hybrigenics web site did not identify any proteins interacting with the N-terminal region of TlpB.
We developed a simple wet-mount-based pH taxis assay in which H. pylori displays pH-tactic behavior that can best be described as directed arcing motility away from the acid source. Using this assay system, we screened the four tlp chemoreceptor mutants for any defects in pH taxis and identified chemoreceptor tlpB as required for pH taxis. Moreover, only tlpB mutants were defective for mouse colonization. Allelic complementation of the ΔtlpB mutant by expression of the WT tlpB gene, cloned in single copy into the rdxA locus, restored both pH-tactic behavior and infectivity for mice, indicating that pH taxis, like motility and urease activity, is an essential factor for colonization of the gastric mucosa. The addition of urea to HCl did not abolish pH taxis by H. pylori, consistent with the well-established role of the pH-gated urease in protecting bacteria while they swim out of the extreme acid. Our findings are also consistent with those of Schreiber et al., who provided convincing in vivo evidence that the orientation of H. pylori in the gastric mucus of gerbils requires a pH gradient (25). Our studies support the notion that pH taxis is dominant over other tactic behaviors and enables colonizing bacteria entering the gastric mucus to travel down natural gradients of hydrogen ions to areas more hospitable for bacterial growth. Moreover, temporal pH sensing and millisecond tactic responses help confine the bacteria within a narrowly pH-defined zone which prevents them from swimming up into the more acidic regions of gastric mucus, where bacteria would be eliminated in the normal turnover of mucus.
In comparison with other epsilon proteobacteria, H. pylori possesses a minimum set of chemoreceptors and lacks orthologs of cheB and cheR which are necessary for the methylation/demethylation adaptive response (2, 31). The TlpB of H. pylori shares no similarity with Tlp proteins from the epsilon proteobacteria in either the N-terminal region or in the putative cytoplasmic pH-sensing HAMP domain. The response of H. pylori to acid differs from that of Serratia marcescens and Pseudomonas aeruginosa, which use both positive and negative tactic responses to remain within an optimal pH zone in capillary tube assays (putatively optimizing the ΔpH component of the proton motive force) (39). It is significant to note that H. pylori bacteria did not swim toward the acid or away from the strong base, as observed with enteric bacteria, suggesting a one-dimensional "negative chemotactic" response. This phenotype also appears to resemble a tar (defective for negative taxis from alkali) mutant of E. coli for which Tsr transduces a one-dimensional pH-tactic response signaled by protonation within the cytoplasmic HAMP domain (32). While the HAMP domain of TlpB exhibits no amino acid similarity with the HAMP domain of Tsr, there is sufficient conservation in charge distribution (basic and acidic amino acids as indicated in Fig. 7) to suggest a cytoplasmic pH-sensing mechanism. This charge distribution is also noted in the TlpB ortholog of the H. mustelae TlpB but not in Tlp proteins of H. hepaticus. Mechanistic studies are in progress to determine how TlpB senses changes in hydrogen ion concentration.
Once highly motile H. pylori bacteria have escaped pH stress, motility decreases or ceases, and a wall of accumulating bacteria is observed in the in vitro assay. Interestingly, the bacterial density of the wall increases as highly motile tactic bacteria pile into the sedentary group comprising the wall. Wall formation is most likely an in vitro phenomenon, since in vivo, diffusion of hydrogen ions would be restricted by gastric mucus, and bacterial density would be lower. This version of pH displacement and recolonization might provide a survival advantage as erosion of epithelial cells due to inflammation and loss of gastric mucus (ulcer formation) would increase local acidity, causing the bacteria to move, but not too far from the nutrient source provided by inflammation. The ability of H. pylori to stop swimming in response to a more neutral pH environment might be relevant to the function of the Cag type IV secretion system, since it has been hard to conceptually reconcile how highly motile bacteria would be able to deploy the type IV pili into epithelial cells without first becoming sedentary. Our findings suggest that the sedentary state might optimize attachment to epithelial cells and activation of the Cag system to promote inflammation (8, 12). Thus, the chemotaxis system of H. pylori may be another example of the remarkable ability of Helicobacter spp. to adapt common physiological systems to optimize colonization and life-long persistence in the apparently hostile gastric mucosa of mammals.
Our findings that TlpB is required for pH taxis in H. pylori and is essential for mouse colonization are in stark contrast to the findings of McGee et al., who concluded that tlpB mutants were not defective for colonization of gerbils and who reported no phenotype associated with chemotactic behavior (20). In their study, animals infected with tlpB mutants exhibited measurable differences in inflammation and recruitment of lymphocytes (20). We obtained the tlpB mutant from David McGee, and it appeared to be pH tactic in our assay, though this strain and the supplied parental SS1 exhibited poor motility (data not presented). We also confirmed that the locus was genetically altered, and DNA sequence analysis indicated that the tlpB gene was indeed deleted as originally described. As pointed out in previous studies, gene duplications (mutant and functional allele) are not uncommon in this species (6) or are second-site mutations in genes that might produce an avirulent phenotype. In our studies, vector-free allelic replacement mutagenesis was used to create all mutants used, and in the case of tlpB, mutants produced in three independent experiments yielded a consistent phenotype. Therefore, allelic complementation of the ΔtlpB mutant by the single-copy insertion of a functional tlpB gene into the rdxA locus provided a critical test of the hypothesis and formally established the requirement of TlpB function for pH-tactic behavior and mouse infectivity. Studies are in progress to resolve pH-tactic differences between these strains.
The chemotaxis system of H. pylori is complex and includes a fused CheAY histidine-kinase/response regulator and three CheV paralogs, each of which contains an N-terminal CheW domain (23) and a response regulator domain of the CheY family (9). It has been suggested that the CheW portion of CheV proteins binds to the Tlp chemoreceptors and possibly modulates phosphorylation of CheA. However, none of the cheV genes can rescue a cheW mutant of H. pylori (23). Mutational analysis of the cheV genes indicated that cheV1 was essential for chemotactic behavior, while the other two mutants displayed WT chemotactic abilities as judged by soft agar assays (23). These findings do not exclude cheV2 and cheV3 in other specialized chemotactic responses. Several groups have reported chemotactic responses of H. pylori strains to bicarbonate, ammonia/urea, bile, and various amino acids (5, 9, 22, 40). Yet, other studies have not been able to reproduce some of these findings (3), underscoring the need for more mechanistic studies of this system. There is also controversy regarding the necessity for a functional chemotaxis system for infections in animal models. For example, initial studies showed that cheW, cheA, and cheY mutants failed to colonize mice (9, 23), whereas more-recent studies showed that these nonchemotactic mutants were able to colonize mice (3, 30). Clearly, much more study is required to identify both the substrates recognized by each chemoreceptor and the specific phosphorelay proteins coordinating each tactic response. Such studies will require development of more-robust nonnutrient assay systems in which single compounds can be tested. We found that H. pylori flagella are easily sheared (e.g., by centrifugation and resuspension of bacteria in other chemotaxis buffer systems), limiting pH-tactic studies to dilutions of bacteria in growth medium. Foynes et al. similarly diluted H. pylori bacteria in a phosphate-citrate buffer in their chemotaxis assays (9). Our studies also demonstrated variations in motility and pH-tactic behavior for tlpD and tlpC mutants, the former demonstrating more arcing and tumbling behavior and the latter displaying hypermotility that accelerated wall formation. Yet, these mutants behaved essentially as the wild type in terms of mouse colonization. Further study will be required to assess the interactions of these chemoreceptors in mediating or amplifying tactic behavior.
We do not know how pH taxis integrates into other acid-adaptive behaviors that include the effects of pH-gated urease (4, 26, 38) and the various transcription-based two-component regulators that control global gene expression, including adaptation to acid stress (7, 33). Studies by Delany et al. indicate that tlpB may be regulated by an orphan response regulator HP1043 (7), while one of the cheV genes appears to be regulated by the nickel response regulator NikR (33). Further study is needed to assess the role (if any) of tlpB in acid stress and, more importantly, to evaluate the extent to which pH sensing and tactic behavior minimize the effects of acid stress and the magnitude of acid-adaptive responses in general.
In summary, we have demonstrated that chemoreceptor TlpB, whose N-terminal periplasmic and HAMP domains are conserved among gastric-colonizing strains of Helicobacter, is essential for pH taxis and for mouse infectivity. The fact that chemoreceptors tlpA, tlpC, and tlpD are dispensable for mouse colonization underscores the importance of acid in defining both the site of colonization by H. pylori and the ability of these bacteria to persist in the gastric mucosa.
We thank Rafael Garduno for assistance with the imaging system, Sander van Zanten for helpful discussions, and Donna Hutchison, Peter Ernst, and Elizabeth Wiznerowicz for the animal studies.
This work was supported by a grant from the Canadian Institutes for Health Research and startup funds from the University of Virginia Health Systems.
- Received 13 September 2005.
- Accepted 18 January 2006.
- Copyright © 2006 American Society for Microbiology


text:
26 July 2014
Whitby regular, Matt Breen from York had a trip to remember when he landed a pb 280lb+ Porbeagle shark. Mistress Charters skipper Rich Cope tells the story of the how the new boat record was beaten.
20 July 2014
What a fabulous day. What a great atmosphere. What a great cause. Three thoughts I had as I drove away from Manor Farm Lakes in Bedfordshire having just attended the 2014 Carping4Heroes event writes Brian Skoyles.
20 July 2014
After Martyn Skoyles had to cancel his carefully laid plans for trip to the River Windrush on the 16 June, the wait proved to be worth it when the rescheduled day produced this cracking 5lb 10oz chub.
13 July 2014
Like many UK carp anglers, our consultant Brian Skoyles looks forward to his regular visits to France each year in search of the prolific fishing that can be on offer. He reports back on a recent two week trip to two venues that certainly didn't disappoint.
17 June 2014
Bob Roberts reveals how this year's finalists got on in the Daiwa Club Match Angler Championship at Sykehouse Fisheries.
23 May 2014
As Brian Skoyles pulled up outside the Hallcroft fisheries café and tackle shop, the carp park was full, as was the café itself. It was time for the draw on this year's Daiwa Dealer Day.
23 May 2014
It is widely regarded that the most prestigious individual event in Irish angling is the 3 day Waterways Ireland Classic festival held in the picturesque town of Enniskillen in County Fermanagh.
12 May 2014
Daiwa consultant Brian Skoyles has just returned from a short but spectacular trip to a new venue called Jay Lake in central France. There with good friend Bill Cottam at the invite of owners Paul and Debbie Smith, Brian and Bill banked 16 carp to just over 53lb in three days, superb fishing by any standards.
05 May 2014
Martyn Skoyles reports back on a recent trip to Texas in search of some spring sunshine, plus hard fighting common and buffalo carp.
05 May 2014
Mike Dearing from Beverley netted himself £1,050 in prize money after landing a 20lb 7.5oz cod on board the Mistress to win the WCSA Spring Festival.


text:
Our 5 Star Piran Meadows Indoor Swimming Pool…
There is something for everyone at the indoor swimming pool in Newquay's 5 star Piran Meadows Resort.
Offering fun filled activities, adult only swim times and swimming lessons under the guidance of trained instructors, you will be able to enjoy the swimming pool experience you are looking for in our 5 star facilities.
Our heated swimming pool is open to the general public, with all of our guests also being able to take advantage of our on-site sauna and steam room whilst on-site; making the most of the luxury offering which only Piran Meadows can provide.
Our Swimming Pool Timetable
Our exclusive Newquay Swimming Pool is open to members of the general public as well as members of our holiday home owner community and any holiday guests who are enjoying a luxury holiday at Piran Meadows.
Anyone living in the area around Piran Meadows who is looking to utilise our heated indoor pool is advised to call before arriving to ensure that the facility is available, as at selected times of the year it will be reserved for the use of anyone staying on-site at our 5 star holiday resort.
However, the pool is generally open seven days of the week with designated timeslots for specific activities. This includes our adult only swim times which is for over 16s only where you will be able to do a few lengths of the pool or just chill out without distractions.
There are also time slots which are allocated for Go Active pool activities which includes activities such as the thrilling sea scooter offering.
Those who aren't very confident in the water or are looking to boost their swimming proficiency are also able to take advantage of the Piran Swim School which is open at specific times of the week with separate classes depending on your skill levels.
You can view our latest Newquay swimming pool open times and activity timeslots by clicking the button below:24th Feb – 1st Mar
The Piran Swim School
We have friendly and highly trained instructors at Piran Meadows who are able to offer swimming lessons to various skill levels, with beginner, intermediate and advanced swimming classes available depending on your ability.
One of the advantage of our swimming lessons is that you will not be expected to pay any upfront fees and also won't be required to sign any contract. This is all about maximum flexibility, giving you the chance to progress at your own pace. We also offer trial lessons so that you can decide whether or not our Piran Meadows swim lessons are right for you with no obligation to sign up thereafter.
Our Swimming Lessons are available to owners on the resort and local residents. They cost just £6 per session with the cost reducing to just £5 per lessons if you purchase a block of ten. These lessons take place during designated time slots highlighted below which are split out by ability level:
- 16:30 – 17:00 – Beginner
- 17:00 – 17:30 – Intermediate
- 16:00 – 16:30 – Advanced
- 09:00 – 09:30 – Beginner
- 09:30 – 10:00 – Intermediate
- 09:00 – 09:30 – Beginner
- 09:30 – 10:00 – Intermediate
- 10:00 – 10:30 – Advanced
Get in touch with a member of our team to arrange your trial lesson by completing our online form or by giving us a call on 01726 860415.
Book Swimming Lessons
Go Active Pool Activities
Piran Meadows enjoys a partnership with holiday booking agent Hoseasons which means that we are able to offer our customers access to their exciting and eclectic Go Active programme.
Many of these activities take place during allocated timeslots in our heated indoor swimming pool. There is something for everyone here of all ages, with snorkelling sessions and even adrenalin fuelled activities including Waterwalkerz.
We also host regular pool parties which are available from £4 per person. Please note that under 8's must be accompanied by an adult.
Join An Exclusive Club
You can enjoy access to our state of the art Newquay swimming pool and the rest of our 5 star facilities here at Piran Meadows Resort whenever you like when you become a member of our exclusive club.
From just £25 per month you will be able to access our complete range of 5 star leisure facilities and activities which includes our indoor swimming pool. Other on-site facilities you will be able to make use of include:
- Air conditioned gymnasium
- Sauna & steam room
- Astro pitch
- Sports simulator
- Children's splash zone & soft play area
- Ten-pin bowling alley
- Astro pitch
In addition to this, you will benefit from 10% off the price of food and drink at our on-site bistro and 10% off any Go Active activities on-site at Piran Meadows.
Alongside individual membership, we also offer Family membership packages from just £50 per month which will help keep your kids happy and occupied during the long school holidays.
Please note that we have a very small number of membership opportunity available, with enquiries dealt with on a strictly first come, first serve basis. If you are interested in becoming a member of our ultra-exclusive club, please contact our team on 01726 860415 or by completing our online form at the bottom of the page.
Membership Enquiry
Contact the Piran Meadows Team
If you would like to contact a member of the team behind our state of the art Newquay Swimming Pool to enquire about availability or request more information about our offering, please feel free to complete the form below.
We are also happy to take requests for membership of our exclusive Piran Meadows club so that you can make the most of our Award Winning facilities whenever you feel like it:
Darwin Escapes keeps the interests of its customers top of mind, so you can rest assured that your data will only be used to help you and will not be shared with external parties unless otherwise requested. For more information about how your data will be utilised, please visit our privacy policy page.


text:
Phil Weston arrived with his girlfriend Maggie Calder, her children and his daughter Jess. He had recently split up with his alcoholic wife, Anne-Marie. Jess and Maggie didn't see eye to eye at all. They fought over minor things.
Phil was stuck in the middle, but got on with Craig and Lucy. Jess continued to try and break up the new family and soon teamed up with Nicola Blackstock. Nicola plotted against Maggie, and was very friendly towards Phil.
But Maggie and Phil's relationship survived. In the end Jess left the village with her mother. Phil was upset about this, but soon got over it. Phil got a job working for Tate Trash.
He left the village in July 2002, a few months after arriving, when he found out Maggie had an affair with Nicola's father Rodney Blackstock.
His portrayer, Mark Jardine, was one of the people who won singing competition Soapstars but the family were so negatively received that they left a few months after arriving.


text:
March 13 to 17 is spring break at Mayville State University. This means that many students headed for home, or maybe to warm and sunny places for rest and relaxation. Others, however, stayed on campus and in town so they could work at their jobs and find other things to do closer to their Mayville State home.
Spring break also means the campus cafeteria is closed, which left those students who stayed on campus or in town on their own for meals. At first glance, this may not seem like such a big deal, but the spring semester is ticking away and financial aid funds may be dwindling for many students as the semester comes to a close.
To help make sure the food needs of these students were covered, Mayville State University Staff Senate announced a food drive on March 7. Donations were collected until Friday, March 10. Faculty and staff stepped up in a big way! Enough food, including cereal, Ramen noodles, granola bars, soups, bread, fruit cups, and much more, was collected to put together 35 over-stuffed grocery bags. Additional donations that didn't fit into the 35 bags meant a mid-week snack boost distribution.
"The generosity and outpouring of support from MSU staff and faculty is truly amazing!" said Misti Wuori, who proposed the food drive. Wuori is vice president of the MSU Staff Senate.
Care packages were left for students to pick up at the campus mail room and distributed on a first-come, first-served basis. They disappeared as quickly as they appeared, and students were grateful for this gesture of personal service that meant they wouldn't go hungry during spring break 2017.
Photo caption: Jessica Amb, Staff Senate president, (left) and Mindy O'Connor, Staff Senate past president, (right) were among the Staff Senators who assembled spring break care packages for Mayville State students.


text:
Will 2016 bring more of the same for Puerto?
Where did 2015 go? Or more to the point, what happened? Not globally, I mean locally, right here in Puerto de la Cruz.
This is the time of year I normally offer a brief summary of the past year's events, happenings, and, of course, a particular favourite of mine, a rundown of the public works operations undertaken. This year is no different; however, the brief summary will be extra brief.
What did happen? The year started with so much promise. The San Telmo project finally managed to drag itself to a successful completion. Is it a success? I'll not comment, I have probably said enough.
What else? There were a lot of promises, all the old favourites, the Bus Station, Playa Martiánez, and let's not forget, Puerto's new port. All this talk and no progress, but then it was election year, so nothing major was ever going to start before May, when we all went to the polls to elect the new team.
How did we fare in Puerto? About as well as could be expected with proportional representation, there were no outright winners. We were left with a coalition. Of the three main parties, ironically, the party with the least votes found itself in the position of power, whilst the party which actually received the most votes found itself in opposition. Makes you wonder why they bother with elections and ask the public who they want, doesn't it? Still, history now, it seems so long ago, the new team at the town hall were quick to act and fulfil all its pre-election promises, if only.
I was optimistic; I allowed them a bit of slack to get their feet well and truly under the table and was then expecting them to come out all guns blazing. Do you believe me? There were a few announce-ments to test the water and, of course, the obligatory photo opportunities, but then you have to make sure you face becomes known, so everyone recognises you.
Apart from that I have never known a council to be so quiet. Not that quiet is necessarily a bad thing, if there is nothing new to say, then why say anything. So often in the past, previous regimes, have made promises, start dates on projects, which never came to fruition, and still haven't. Talk for talk sake has left residents of Puerto in a state of apathy and there is a, I'll believe it when it happens, feeling when it comes to new projects.
What is stalling all these worthy projects? Money, obviously, or the lack of it, a problem not unique to Puerto; the whole island suffers because of it.
Surely something interesting must have happened, apart from the usual festivals which happen year to year that is. I know, extra parking spaces have been created at the old bus station, hardly earth shattering, but more so than you think. Stand there as a couple of cars go through and the earth tremors. I would love to get access to the old underground car park, long condemned, to see just what is holding everything up, but then again perhaps I wouldn't. Oh, and the new council has continued with the 'Underground Wheelie Bin' project, I had expected better of them, but maybe they were already committed.
Enough about 2015, it's all history now, so let's look forward to 2016, I'm positive it will be a more progressive year for the town. We know the next project, it has already been announced, the 'Concrete Slab Makeover' moves on to Calle Quintana. A pedestrian walkway, so, at least, no loss of parking spaces, just the promise of eight months, which will more than likely be a year, of upheaval to shop owners, residents and tourists alike.
All in the name of technology as fibre optic cables are installed, however, I am sure it will look much better when it's completed and the grey slabs will blend in with the rest of the town. No firm start date yet, just 'after Reyes', though I should think they will wait until after Carnival, the logical thing to do.
Let me quickly mention the 'big three' again and then I am signing off on what must be my shortest article, I apologise, but for once I find myself lost for words, or perhaps bereft of anything practical to say, would be a more apt description.
The Bus Station, all the 'correct' paperwork has been submitted, simply waiting for the green light from central government, which could be a long time coming given the fiasco of the December general election.
Playa Martiánez is waiting for the Coastal Department to allocate funds to it in its annual budget. Although now it has been announced the project will be completed in phases, the usual ploy if you can't afford to do it in one go, so maybe we will see some activity during 2016. Puerto's New Port, still talking to possible private investors was the last I heard.
That's it, Happy 2016.


text:
Marine renewables are technologies that generate electricity from tide and wave motion. They produce electricity without greenhouse gases, and could provide economic benefits for the UK. However, the technologies have been slow to develop, despite previous projections of growth. This POSTnote examines the causes of this delay and how the sector might develop.
Documents to download
Energy Use Behaviour Change (456 KB, PDF)
Behaviour change concerning energy use is an emerging area of research that has important implications for policy. This note introduces the factors and interventions that can influence behaviour. It also discusses the behavioural aspects of the Green Deal and the smart meters programme.
Documents to download
Energy Use Behaviour Change (456 KB, PDF)
Related posts
Key infrastructure areas such as transport, energy, water and telecoms are vital to society and the economy. Evidence suggests that climate change is increasing the frequency and severity of extreme weather events, posing a risk to infrastructure systems. This POSTnote looks at the main climate-related risks to the UK's economic infrastructure, measures to reduce these risks and the main challenges to implementing resilience measures.
According to global climate and economic models, removing greenhouse gases from the atmosphere will be necessary to limit global warming to 1.5˚C. Among Greenhouse Gas Removal (GGR) techniques, these models assume that Bioenergy with Carbon Capture and Storage (BECCS) could play a prominent role. This POSTnote summarises why BECCS has been included in the models, outlines the challenges and trade-offs of deploying at scale, and considers policy options for supporting its development.


text:
Are you suffering from vitamin and mineral deficiencies? If you're newly diagnosed with celiac disease or seem to be experiencing continued symptoms, your small intestine may still be suffering damage.
A compromised gut causes a lot of health problems and one of the major problems it causes is an inability or reduced ability to absorb much-needed nutrients.
- Calcium
- Folic Acid
- Iron
- Magnesium
- Vitamin A
- Vitamin B6
- Vitamin B12
- Vitamin D
- Zinc
In a study done by VU University Medical Centre (Amsterdam), "almost 90% of celiac disease patients were found to be deficient in at least one or more of the assessed nutritional parameters, and half of patients were deficient for two or more nutritional serum variables."
In fact, 17% of the untreated celiacs in this study were categorized as malnourished. So it's important to be aware of your potential for nutritional deficiency as well as what deficiencies are most common.
This study tested for 7 nutrients and showed the following results. Be mindful that sun exposure can strongly influence vitamin D levels, and this study was conducted in the Netherlands.
- Zinc – 67% of participants were deficient
- Ferritin – 46% were deficient (a protein that stores Iron)
- Vitamin B12 – 19% were deficient
- Vitamin B6 – 14% were deficient
- Vitamin A – 8% were deficient
- Vitamin D – 5% were deficient
In addition, 25% of participants had iron-deficiency anemia.
It's not uncommon for adults and children with celiac disease to have low bone density (osteopenia) and even osteoporosis.
Bones are the frame of your body, so something like a broken hip or a cracked vertebrae can have a tremendous impact on your quality of life. If you have any concerns at all, be proactive and talk with your doctor.
If you're celiac and have recently broken a bone and/or are over age 50, it's a good idea for you to ask your doctor for a bone density test.
Celiacs also tend to have vitamin D and magnesium deficiencies, which are needed to absorb calcium, exacerbating the situation. So if you need more calcium, you may need more magnesium and vitamin D.
How do you know if you have a health issue stemming from a lack of proper nutrition? Ask your doctor to test your vitamin and mineral levels!
Many celiacs are months or years into their gluten free lifestyle and still struggle with significant health issues. If you fall into this category, work with your doctor to see if you're deficient in critical nutrients.
Some minor dietary changes may be able to make a world of difference in how you feel!
You can find the study referenced above here.


text:
Deepika Samson to play lead in UTV's show
Deepika Samson has been finalized to play lead in UTV and Star Plus' show, which is a remake of Kalangal...
A little birdie tells us, "The strong minded, independent, working woman in the character 'Abhi', will be essayed by Deepika Samson."
Deepika made her debut in TV with Imagine's Devi. She is presently seen playing the role of Rekha in Zee TV's Agle Janam Mohe Bitiya Na Kijo.
As reported by Tellybuzz, the male lead, a charmer, who will go on to have a grey shade will be played by Rohit Khurana. Noted actor Rohini Hatangini, Aishwarya Narkar and Anjan Srivastava have been finalized to play key roles.
We tried calling Deepika, but she remained unavailable.
All efforts to get in touch with Mayuree Chaddha Fiction Head, UTV were futile.


text:
Grow Your Own At Home!
Fast Plain Box Shipping.
We ship to the US & Canada.
Posted on 26th Oct 2015
It's a big question that we see quite a lot: what kinds of fish should people put in a cutting-edge aquaponics garden system?
The answer really depends on a lot of things – on the specific setup, the climate you're in, state or federal laws on wildlife, and much more. But here are some super-popular types of fish that you can put into an aquaponics reservoir.
Tilapia is one of the most common solutions for hydroponics. It's a less fatty fish, and one that often produces a high level of synergy with the plant environment. It's also popular fish for the dinner table. But in some cases, rules on tilapia prohibit growers from using this fish because it's an invasive fish species.
Goldfish and Koi
You can't eat these types of fish, but they do look good, and they are also a good choice for aquaponics gardening. They have a lot of plant synergy, and they are easier fish to manage over time.
Catfish is another option, especially for reservoirs of a larger size. Unlike goldfish and koi, catfish are edible. In fact, a lot of people like to eat catfish. Also, they're often seen in ornamental and decorative ponds, and so it makes sense that they would often be seen in aquaponics as well.
Carp are bottom feeders - and as a result, some people think of them as "dirty" fish. However, there is a market for edible carp, partly because of their use around Jewish holidays. Also, carp can be comfortable in a hydroponics reservoir environment because of the way they live in the wild.
Yes, these popular fisherman catches can also be seen in aquaponics environments. They're another kind of edible fish that can thrive in the gardening setup and be farmed somewhat easily.
Other Types of Critters
In addition to fish, growers can also use creatures like shrimp and crayfish. These smaller marine animals can also function in an aquaponic environment. In some cases, they help to keep the reservoir cleaner. However, growers have to look at possible predatory behavior and the food chain before introducing various types of non-fresh green animals to the hydroponics space.
These are just some basic tips for aquaponics gardeners. To find out more, check out Dealzer's website and everything that we offer for aquaponics and hydroponics and all other kinds of cutting-edge innovative solutions.


text:
Juniata houses chapters for two national leadership honor societies.
National Society of Leadership and Success
NSLS is the nation's largest leadership honor society. Students are nominated through the college as early as their sophomore year. To accept membership, students must attend Orientation, Leadership Training Day, three Speaker Broadcast, and three Success Networking Team Meetings.
Interested in learning more about NSLS at Juniata? Contact firstname.lastname@example.com
Omicron Delta Kappa
ODK is one of the country's preeminent leadership societies in the country. Membership in ODK is granted to those who demonstrate achievements in one of five phases of campus life:
- Scholarship
- Athletics
- Campus or community service, social and religious activities, and campus government
- Journalism, speech, and mass media
- Creative and performing arts
Students must be juniors or seniors who rank in the top 35% of their class and are expected to demonstrate the five ODK ideals of scholarship, service, integrity, character, and fellowship.
Eligible students will receive a request to apply in the Fall of their junior or senior year. Tapping and induction of selected students occurs in the Spring Semester.
Interested in learning more about ODK at Juniata? Contact email@example.com


text:
This article provides information on how to get rid of candida naturally.
We all know that yeast is good for muffins, but what about your "muffin"?
You would be surprised to know that we already have yeast in our bodies. We actually need it. It comes in the form of a fungus known as candida which lines various areas of the digestive system. In very small amounts, it's pretty useful in helping with digestion and allowing the body to metabolize the nutrients from the food that we eat.
But just like any other fungus, it can grow rapidly and cause a lot of problems if the conditions are right. And one of the worst manifestations of this fungal outbreak is in the genital area.
That's right. Yeast infection can happen in your muffins. The medical term for this phenomenon is called genital candidiasis. And although it's not a fatal disease, it can cause all sorts of problems:
- Difficulty with digestion
- Loss of focus
- Lowered Self-Esteem
- Hives, rashes and psoriasis
- Lupus
- Diarrhea
And these are just to name a few. Because the infection starts out in the digestive system, your symptoms will start out in the tummy. Instead of getting nourishment from food, it all goes to feeding the growing yeast in your digestive system.
Risk Factors
Since all of us already have the yeast in our bodies, it's the lifestyle choices that we make that determine if the yeast in our bodies stays in control or not. Some of the things that can promote the growth of yeast include:
- Excessive alcohol intake
- Oral Contraceptives
- Constant stress
- Antibiotics
You may consider this as somewhat normal things for a young individual with an active lifestyle. But constant work and drinking without any rest can cause the good bacteria in your stomach to die off. These bacteria are essential to keeping the yeast in check. When these security guards go out, the yeast grows and begins showing in various areas of your body.
About 75% of all women have reported a yeast infection at least once in their life. This could have shown on any part of their bodies. But the most common and frightening place to show up would be right in the genitalia.
If you experience itching right in between the legs and the tips of your fingers, you might want to check with a doctor to see if you have elevated levels of yeast in your blood. High amounts of yeast in the blood will show up through a routine blood check. Lab technicians are trained to read the various levels of materials in your blood and will let you know if you're harbouring more yeast than you're supposed to.
Another way to detect a yeast infection (before it shows up) is through a urine and stool sample test. Because yeast is a growing organism inside your body, it will also expel its own waste materials that go out with your own. A test that shows high levels of D-Arabinitol is a sure indicator of excessive yeast in the body.
Treating Candida
The first step in taking care of candida is to stop the growth of the yeast. That means you have to take a break from your current lifestyle. On top of that, you are also going to need supplements to inhibit the fungal growth.
The first thing that goes out of your diet is sugar. This is because sugar is known to promote the growth of yeast as it provides energy for the fungus to multiply. Instead of taking in sweets, a low-carbohydrate diet has to start so that you stop feeding your fungus.
To stop the yeast growth, you can take various substances to kill the fungi and restore the good bacteria in your digestive system. A number of medications are designed just for that purpose.
Some of the more popular forms of candida supplements include Caprylic Acid and Diflucan. What these substances do is that they immediately attack the grown fungi in your digestive system and break it down into a form that you can eliminate from your body.
In contrast to taking supplements for candida, you can also learn how to get rid of candida naturally through a few simple changes to your lifestyle.
How to Cure Candida Naturally
But besides taking over-the-counter solutions to your yeast infection, you can also try some natural remedies for candida that don't cost as much but will still achieve the same effects.
Plenty of natural food items will aid in killing the fungal infection without the help of medicines. These can be considered as natural cures for candida. You want to focus on eating fungi-killing items such as grapefruit seed extracts and berberine. The idea here is to destroy the built-up yeast in your body after starving it.
You don't necessarily have to starve yourself by avoiding meals altogether, but depriving the fungi of sugar will certainly reduce its resistance to caprilyc acid and grapefruit seed extracts. That also means taking out sweeteners, both natural and artificial, for the mean time.
Processed foods that are full of gluten and preservatives are also known to feed the yeast, so avoiding them for a few months will be a strict rule of your natural remedy. The more natural your food choices, the more useless they'll be for feeding the yeast in your body. And when the yeast cannot use what you eat, it ends up helping you improve your immune system and replenish the good bacteria in your gut.
Another part of your natural candida treatment plan is fiber. This is closely related to the digestive system as it helps you eliminate all the unused food material in your gut. And since you won't be needing any of the fungus you have stored in there, increasing your fiber intake will most certainly help you get rid of the growth at a faster rate.
You want to take soluble fiber because you're guaranteed these substances do not have any preservatives. Some great examples of these are pectin, flaxseeds and guar gum. These are prescribed by experts as a natural candida cure. Be sure to accompany these fiber forms with a lot of water to help wash out infestations all around your body. The yeast may be showing up "down there" but that doesn't mean it's not in other places in your body.
Finally, the last natural cure for candida is garlic. What first looks like something you throw into the frying pan is actually the most feared predator of fungi in the gut. This is because garlic contains concentrated amounts of alliin, allicin and S-allylcystiene. These substances are known to poke holes in fungal growth, immediately starting a decline in its development.
To use garlic in your diet, be sure to crush your cloves before chopping them to release all of its fungi-dissolving properties. About 4 or 5 cloves will show improvements within weeks. You can also consume natural garlic tea of you don't want to include garlic in your cooking. Or you can always take the rough road and consume the actual cloves without cooking them first.
Natural Remedies: Healing the Tummy
While you're kicking out the unwelcomed guests in your digestive system, you have to start inviting some ideal tenants in the space that you're creating for them. These are known as "probiotics". These are good bacteria that help the intestines break down your food and keep digestion going smoothly.
The idea of consuming bacteria may not be a welcome thought to someone suffering from a fungal infection, but eating the right kinds of food will help make the process enjoyable. Many fermented food items can serve as a great source of good bacteria.
Some great examples to choose from are kimchi (spicy fermented Korean vegetables), yogurt (tangy fermented milk) and sauerkraut (fermented chopped cabbage). These popular food items will both taste great and give you a good refill on the good bacteria in your stomach.
In totality, it can take a few months for you new diet and lifestyle to completely eliminate the yeast in your body. This is because you still need to starve them before you take them out. Since fungi are especially keen on surviving in the harshest conditions, it will take some time to make your gut an inhospitable area for them.
Some treatment cases can take half a year, some take half of that. Depending on how you change your lifestyle and control your food intake, your recovery is almost always guaranteed.
Psychological Support
Although you're not dealing with cancer or anything life-threatening, dealing with a new diet, lifestyle and a growth in your private areas can be something that is hard to deal with.
On top of the mental strain, a certain part of then fungal cure is a "dying out" process that is exhibited by the worsening of the symptoms right in the middle of treatment. At this point, constant care should still be taken as well as vigilant efforts to maintain the diet. It's very easy to promote the growth of a fungus, but it takes some time to get rid of it.
Aside from this "dying out process" you should also focus on taking it easy with the treatments. Doubling your intake of garlic and caprylic acid is not going to make things heal faster. These things take time but will be worth the trouble after a while.
Finally, when the fungus has been removed from the system, the infections, swelling, growths and even marks on the genitals eventually disappear. What's left is a lingering lesson on how to keep yeast on muffins and not on your "muffin".
>>Click Here To Get Rid Of Candida Naturally<<


text:
Then as Sue Neales in the Mercury comments, some might see positives in the release of what amounts to a government guarantee for Gunns regarding sovereign risk concerning an aspect of the deal for the supply of timber to the proposed pulp mill. But why unprompted disclosure four months after the guarantee was signed?
"(And what about an)explanation about why 25 documents -- held within the Department of Economic Development relating to the Government's well-advanced plans to fund the private pulp mill's $65 million water supply pipeline -- could not be made publicly available. A Freedom of Information request to obtain this information this week received a blanket refusal on the grounds the information was commercial-in-confidence and formed part of protected information provided to the Government's inner Cabinet.
Increasingly around Australia, both arguments are being used as a form of obfuscation by governments seeking to prevent the public using FOI laws to discover what is actually going on behind the thick walls and closed doors of executive government. The problem is, as Michael McKinnon, Australia's leading FOI media exponent told a packed lecture at the University of Tasmania on Thursday, these excuses are usually used as a cover-up.
"Where you get this sort of government secrecy (in relation to FOI requests), you can almost always guarantee that there is a sweetheart deal going on," McKinnon said bluntly this week."


text:
Walk With Ease
Walk With Ease is a free program designed to help those with arthritis better manage their pain, but it is also ideal for anyone who wants to make walking a habit! This six-week course will begin Jan. 13, 2020, and run every Monday, Tuesday, and Thursday through Feb. 20, 2020, from 5:15 p.m. to 6:15 p.m. Meet at the Burns High School Library. Walk With Ease includes a warm-up, stretching, walking, and a cooldown. No registration is required. For more information, call Suzanne at 541-573-8614.


text:
A record breaking 2018 and our plans to make 2019 even better
Posted on 21-01-19 by Admin
We are certainly living in interesting times right now. Whether you are in favour of Brexit or against it, one thing I think that is unifying the British population is our collective dismay at how poorly our elected politicians are handling the process, whichever…READ FULL POST
Posted on 03-09-18 by Admin
Friday was a fantastic day for all at Clear Edge. Off we went to Horsforth Park for our annual sports day, raising all we could for The Alzheimer's Society. We piggy backed, ran, jumped and even rolled (sorry Mark) our way through the…READ FULL POST
Celebrating 7 years of Clear Edge
Posted on 22-08-18 by Admin
We are celebrating our 7th Birthday! We've had lots of excitement in the office today. This morning, each team member was given a birthday balloon with a hidden prize inside, ranging from lunch on the directors to crates of beer and wine! After…READ FULL POST
Posted on 28-03-18 by Amber Mcdonald-Malone
We are pleased to announce that this year, we have selected to support the Alzheimer's Society as our chosen charity of the year. The charity itself work tirelessly to transform the landscape of dementia forever. Dementia is the biggest health and care challenge…READ FULL POST
6 crucial steps to securing talent | Consumer goods recruitment
Posted on 28-02-17 by Terry Batty
In today's employment marketplace there is a genuine shortage of top talent available. This means that the best candidates can afford to be very selective about which companies they join and which offers of employment they decide to accept. Terry Batty, Clear Edge Director…READ FULL POST
Sam Gray joins our Consumer Durables Recruitment Team
Posted on 20-02-17 by Terry Batty
We're delighted to announce that experienced recruitment specialist, Sam Gray, has joined Clear Edge Search & Selection. He specialises in the Homewares and Gift categories and will further strengthen our highly successful consumer durables team. Sam joined us from a management position at his…READ FULL POST
Clear Edge shortlisted in Best Niche/Specialist category at 2016 Recruitment Business Awards
Posted on 12-08-16 by Terry Batty
We are thrilled to announce that Clear Edge has been shortlisted for a Recruitment Business Award in the Best Niche/Specialist Agency category. The awards are run by The Drum and recognise the best Recruitment Businesses (and advertising agencies that work with the recruitment…READ FULL POST
A lack of fresh commercial talent in the FMCG sector and how to attract it
Posted on 05-05-16 by Terry Batty
There's a common theme our specialist consultants are hearing when taking vacancies from clients. It sounds something like this… "We'd like someone who is up and coming. A bright talent for the future that can develop further within our business but they already have…READ FULL POST
The charmed life of an Interim Manager? Balancing the risks of contract work with the potential rewards.
Posted on 01-03-16 by Terry Batty
At some point in your career, you've probably considered becoming self-employed. Apart from the financial benefits and flexibility this allows you, it also gives you the chance to explore a number of different sectors and companies that would not be possible as a…READ FULL POST
Rising consumer confidence keeps non-food job market buoyant
Posted on 22-02-16 by Terry Batty
From talking to our clients and monitoring market trends, one thing became clear in 2015 – consumers are willing to splash out on life's little luxuries again. This of course is great news for anyone employed in the non-food consumer goods sector, or…READ FULL POST
Search our Updates:
- Filter by category:
- Blog
- Drinks
- Food
- Non-Food
- Operations, Technical & Supply Chain
- Retail
- Sales & Marketing
- Team News
- Uncategorised
For even more of the latest FMCG vacancies, news, insights & recruitment tips...Follow us on LinkedIn
Stay Updated
Keep up to date with the news you need. Please send me:


text:
Our Guarantee!
Every product at PurpleTulsi.com is backed by our guarantee. If you don't like any item - return or exchange within 7 days for a full refund! We even arrange free pickups!
Lord Ganesha symbolises wisdom & removes all Vastu Dosh. God Ganesh also known as Ganpati. Apart from being worshipped, this amazing showpiece can be placed in your living room or in your office. It is considered to provide a lot of positive energy and vibes of courage, hope, power, knowledge, intellect and devotion.åÊ
Size -åÊåÊåÊåÊåÊåÊåÊåÊåÊ W x H x D: 14.0 x 8.0 x 17.0åÊ(cms)
Material - åÊåÊåÊ Brass
Weight -åÊåÊåÊåÊåÊ 1740 g
Delivered in 10-15 Days.
Returns Policy
You may return most new, unopened items within 7 days of delivery for a full refund. We'll also pay the return shipping costs if the return is a result of our error (you received an incorrect or defective item, etc.).
You should expect to receive your refund within four weeks of giving your package to the return shipper, however, in many cases you will receive a refund more quickly. This time period includes the transit time for us to receive your return from the shipper (5 to 10 business days), the time it takes us to process your return once we receive it (3 to 5 business days), and the time it takes your bank to process our refund request (5 to 10 business days).
We can ship to virtually any address in the world. Note that there are restrictions on some products, and some products cannot be shipped to international destinations.
When you place an order, we will estimate shipping and delivery dates for you based on the availability of your items and the shipping options you choose. Depending on the shipping provider you choose, shipping date estimates may appear on the shipping quotes page.
Please also note that the shipping rates for many items we sell are weight-based. The weight of any such item can be found on its detail page. To reflect the policies of the shipping companies we use, all weights will be rounded up to the next full pound.


text:
I have kind of a problem with refunds. When a payment is refunded Curve marks it as refunded and the originally paid amount is not visible anymore. But the reality is that in my credit card statement I will see a charge and a refund, probably with different dates, and it is difficult to reconcile. Couldn't be possible to keep, for information purpose only, the original amount for refunded transactions? Original amount, payment date and refund date would be important information to keep.
Other related problem is that there is no way to look for refunded transactions using the filters. The amount is useless since there is no amount any more, and maybe you want to see what refunds you got, not just checking one specific, so a filter for "refunded transactions" would be very useful.