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NadA is a novel vaccine candidate recently identified in Neisseria meningitidis and involved in adhesion to host tissues. The nadA gene has been found in approximately 50% of the strains isolated from patients and in three of the four hypervirulent lineages of non-serogroup A strains. Here we investigated the presence of the nadA gene in 154 meningococcal strains isolated from healthy people (carrier strains). Only 25 (16.2%) of the 154 carrier isolates harbored the nadA gene. The commensal species Neisseria lactamica was also found not to have the nadA gene. Eighteen of the carrier strains belonged to the ET-5 and ET-37 hypervirulent clusters, indicating that only the 5.1% of the genuine carrier population actually harbored nadA (7 of 136 strains). Five of the seven strains harbored a novel allele of the nadA gene that was designated nadA4. The NadA4 protein was present on the bacterial surface as heat-stable high-molecular-weight oligomers. Antibodies against the recombinant NadA4 protein were bactericidal against homologous strains, whereas the activity against other NadA alleles was weak. In conclusion, the nadA gene segregates differently in the population of strains isolated from healthy individuals and in the population of strains isolated from patients. The presence of NadA can therefore be used as a tool to study the dynamics of meningococcal infections and understand why this bacterium, which is mostly a commensal, can become a severe pathogen.
Neisseria meningitidis, a gram-negative diplococcus that is able to colonize the upper respiratory tract of humans, is a major cause of meningitis and septicemia. In most industrialized countries, even if about 10% of the human population is colonized by the bacterium in endemic periods, only 1 to 3 individuals per 100,000 people develop disease (13). Therefore, N. meningitidis should be considered a commensal which on rare occasions becomes a very dangerous pathogen. The reasons why meningococcus can be a commensal and a pathogen at the same time are poorly understood (23).
Meningococci are classified in serogroups based on the chemical composition of the polysaccharide capsule. Serogroups A, B, C, Y, and W-135 are associated with disease. Most of the meningococcal strains isolated from invasive disease have been classified by multilocus enzyme electrophoresis (4) into hypervirulent lineages (electrophoretic types ET-37 and ET-5, cluster A4, lineage III, and subgroups I, III, and IV-1) or by multilocus sequence typing (MLST) (17) into sequence type complexes (ST-11, ST-32, ST-8, ST-41/44, ST-1, ST-5, and ST-4). Meningococcal carrier populations are much less defined, and they include some proportion of strains which belong to hypervirulent clusters that apparently are identical to the strains isolated from patients. However, most strains normally isolated from healthy individuals are rarely able to cause the disease (14).
Meningococcal carriage is believed to be the natural reservoir of strains responsible for outbreaks. The appearance of strains belonging to an hypervirulent cluster correlates with increased disease frequency, and meningococcal carriage may increase during outbreaks (up to 50%) compared to the carriage during endemic periods (average, 10%) (2, 19). Meningococcal carriage has been shown to be an immunizing event both in children and in adults (3, 10) and is able to induce a bactericidal response. However, the efficacy of immunization for preventing disease is controversial since it has also been shown that carriage cannot induce protection against colonization and invasion (1). Carrier strains are genetically quite diverse compared to disease-associated strains (5, 14). The polysaccharide capsule, which is the most important pathogenicity factor of meningococci, is frequently missing in carrier strains, and this may partially account for the difference in pathogenicity (5, 8). While capsule switching could temporarily occur as a consequence of phase variation (20), the absence of the capsule is often due to the absence of the whole capsule operon (6). Along with the hypervariability of serotype and subserotype antigens, this is the main cause of the inadequacy of the conventional serological markers for tracing the fate of meningococcal isolates, particularly carrier strains (1). In this work we studied the presence and molecular features of NadA, a new adhesin, potential virulence factor, and vaccine candidate recently identified in N. meningitidis, in a large panel of carrier strains obtained from various sources and in the commensal species Neisseria lactamica. The nadA gene is known to be present in approximately 50% of meningococcal isolates and is more frequently associated with strains which belong to hypervirulent clusters. It is always present in members of three of the four major non-serogroup A meningococcal hypervirulent clusters, namely, the ET-5 complex, the ET-37 complex, and the cluster A4, whereas it is never present in lineage III strains. NadA is an adhesin which exhibits homology with a family of proteins involved in invasion and pathogenesis. Its sequence is unusually well conserved, and only three alleles (alleles 1, 2, and 3) have been identified. Allele 1 is harbored by all of the ET-5 strains tested so far, whereas alleles 2 and 3 are present mainly in strains belonging to the ET-37 complex and A4 cluster and also in strains not belonging to any hypervirulent cluster. Our analysis showed that the nadA gene is underrepresented in carrier strains and that in a subset of these strains there is a new allele that we designated allele 4 (nadA4).
Bacterial strains, culture conditions, and chromosomal DNA isolation.A total of 154 carrier strains of N. meningitidis and 18 strains of N. lactamica were selected for analysis. The meningococcal strains came from six different countries (Norway, People's Republic of China, United States, Chile, Iceland, and Oman), whereas the N. lactamica strains came from England and Oman. The years of isolation ranged from 1972 to 2000. Strain 2996, an example of a clinical isolate harboring allele 3 of nadA, and NGH38, a strain that did not contain the nadA gene, were used in functional assays as positive and negative controls, respectively (7). In order to extract chromosomal DNA, bacteria were grown overnight at 37°C in the presence of 5% CO2 in gonococcus agar supplemented with Kellog's solution (0.22 M d-glucose, 0.03 M l-glutamine, 0.001 M ferric nitrate, and 0.02 M cocarboxylase) (Sigma Aldrich). A starting culture of each strain at an optical density at 600 nm of 0.05 was grown to the stationary phase in 5 ml of gonococcus medium. Then 1.5 ml was pelleted by centrifugation at 16,060 × g for 15 min, and chromosomal DNA was prepared by using a Macherey-Nagel NucleoSpin tissue kit according to the manufacturer's protocol. Escherichia coli DH5α and BL21(DE3) were used as a cloning strain and an expression host, respectively, and were used as indicated by the supplier (Invitrogen).
PCR and nucleotide sequencing.PCR amplification of the nadA gene was performed with about 10 ng of chromosomal DNA by using primers located upstream and downstream of the nadA gene coding region. The forward primer was A1 (GTGGACGTACTCGACTACGAAGG), and the reverse primer was B2 (CGAGGCGATTGTCAAACCGTTC). The Invitrogen Accuprime Taq DNA polymerase system was used. The PCR conditions were as follows: 30 cycles of denaturation at 95°C for 30 s, annealing at 56°C for 30 s, and extension at 68°C for 1 min. The amplified fragments were purified with a QIAquik PCR purification kit (QIAGEN) and were sequenced with an ABI 377 automatic sequencer (Applied Biosystems). A sequence analysis was performed by using the A1 and B2 primers used for gene amplification and primers 1 (TATGTAAACAAACTTGGTGGGG), 2 (GAAATAGAAAAGTTAACAACCAAGTT), 3 (GACATCAAAGCTGATATCGCTAC), 4 (TTTCGAGGTGGCGCGTTCGGG), 5 (GTAGCGATATCAGCTTTGATGTC), and 6 (CTTGGTTGTTAACTTTTCTATTTC). Sequences were analyzed by using the EditView program (Perkin-Elmer), a GeneJockey II sequence processor (Biosoft), MacBoxshade, and the GCG Wisconsin package.
Western blot analysis.Total cell extracts and outer membrane vesicles (OMVs) were prepared as described previously (7, 22). The protein concentration in each sample was evaluated by Bradford analysis. Equal amounts of total proteins and OMVs were boiled for 10 or 30 min, either alone, in the presence of the zwitterionic detergent Triton X-100 at a concentration of 2%, or in the presence of the anionic detergent sodium dodecyl sulfate (SDS) at a concentration of 2%. Samples were subjected to SDS-polyacrylamide gel electrophoresis on 5 to 15% Bio-Rad polyacrylamide gels, transferred onto nitrocellulose transfer membranes (Schleicher & Schuell), and hybridized with a 1:200 dilution of an anti-NadA4 mouse polyclonal antiserum. The blots were incubated with a 1:2,000 dilution of the DAKO anti-mouse peroxidase conjugate secondary antibody, and the signal was developed with a Bio-Rad Opti-4CN substrate kit.
nadA4 gene cloning and NadA4 protein expression and purification.The nadA4 gene (encoding amino acids 28 to 268) was amplified by PCR by using chromosomal DNA of strain NGE28, forward primer 961-13F (CGCGGATCCCATATGCCACCGCTGACGAAATTG), and reverse primer 961-C REV (CCCGCTCGAGACCCACGTTGTAAGGTTG) (an NdeI restriction site in the forward primer and an XhoI restriction site in the reverse primer are underlined). The PCR conditions were as follows: five cycles of denaturation at 94°C for 30 s, annealing at 52°C for 30 s, and elongation at 68°C for 1 min and 25 cycles of denaturation at 94°C for 30 s, annealing at 65°C for 30 s, and elongation at 68°C for 1 min. The PCR was performed by using 10 ng of chromosomal DNA and High Fidelity Taq DNA polymerase (Invitrogen). The amplified DNA, after purification and NdeI/XhoI digestion, was cloned into the pET21b+ vector (Novagen). The plasmids were transformed into E. coli BL21(DE3) to express the proteins as COOH-terminal histidine fusions. Recombinant proteins were expressed and purified as previously described (18).
Dot blotting.Aliquots (10 μl) of total extracts of each strain were boiled for 10 min and spotted onto nylon membranes (Boehringer Mannheim). The DNA on the membranes were cross-linked by 2 min of exposure to UV light, and the membranes were stored at room temperature. For probe preparation, the nadA gene was amplified from strain 2996 by using inner primers 961-F (GCCACAAGCGACGACG) and 961-R (CTCGTAATTGACGCC) and Platinum High Fidelity Taq DNA polymerase (Invitrogen). The PCR conditions were as follows: 30 cycles of denaturation at 94°C for 30 s, annealing at 52°C for 30 s, and elongation at 68°C for 1 min. After purification with a QIAquik column (QIAGEN), the DNA (∼350 ng) was labeled with digoxigenin by using a DIG High Prime DNA labeling and detection starter kit II (Roche). Prehybridization, hybridization, and detection of digoxigenin-labeled nucleic acids were performed as recommended by the DIG system user's guide (Boehringer Mannheim).
Binding assay.Chang epithelial cells (Wong-Kilbourne derivative, clone 1-5c-4, human conjunctiva) were maintained in Dulbecco modified Eagle medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 15 mM l-glutamine, and antibiotics. Cells were nonenzymatically detached from the support by using cell dissociation solution (Sigma), harvested, and resuspended in RPMI medium supplemented with 1% FBS. Then 1 × 105 cells were mixed with either medium alone or three concentrations (175, 43, and 10 μg/ml) of histidine-tagged NadA3, NadA4, or GNA2132 proteins and incubated for 30 to 40 min at 4°C. After two washes with 1% FBS in phosphate-buffered saline, cells were incubated with mouse polyclonal antiserum against each protein for 1 h on ice. After two washes, the preparations were incubated on ice for 30 min with R-phycoerythrin-conjugated goat F(ab)2 antibody to mouse immunoglobulin, and the cells were finally analyzed with a FACS-Scan flow cytometer. The mean fluorescence intensity for each population was calculated.
Serum bactericidal assay.Twenty micrograms of purified recombinant protein was used to immunize 6-week-old CD1 female mice (Charles River Laboratories) intraperitoneally (four to six mice per group). Immunization was performed on days 0, 21, and 35 by using Freund's adjuvant or a solution containing 3 mg of aluminum hydroxide per ml. Blood samples for analysis were taken on day 49. Serum bactericidal activity was evaluated as described previously (7) by using baby rabbit serum (CedarLane) as the complement source. The serum bactericidal titer was defined as the serum dilution that resulted in a 50% decrease in the number of CFU per milliliter after 60 min of incubation compared with the number of CFU per milliliter at time zero.
Distribution of NadA among N. meningitidis carrier strains and N. lactamica.We used a panel of 154 carrier strains isolated from healthy individuals; 74 strains were nongroupable, 52 strains belonged to serogroup B, 6 strains belonged to serogroup C, 1 strain belonged to serogroup K, 1 strain belonged to serogroup I, 4 strains belonged to serogroup W-135, 6 strains belonged to serogroup Y, 3 strains belonged to serogroup X, and 7 strains belonged to serogroup Z. By using PCR and dot blotting, 25 (16.2%) of the 154 strains were found to be positive for the nadA gene. This frequency is much lower than the frequency observed for strains isolated from patients (approximately 50%). The presence of NadA in the different strains is shown in Table 1. The genes in all positive strains were sequenced, and the sequences were compared to the sequences of the three previously described nadA alleles. The strains harboring NadA are described in Table 2. Allele 1 was found in all strains belonging to the ET-5 hypervirulent cluster as previously described for ET-5 strains, isolated from patients (7). Allele 2 was found in all strains belonging to the ET-37 hypervirulent cluster and in the 16747 strain (ST-189). Strain 2022 carried a nadA2 gene interrupted by the insertion sequence IS1301 (11), as sometimes occurs in clinical isolates. The remaining five positive strains (3.2% of the total), which did not belong to any hypervirulent cluster, had a new nadA allele that we designated nadA4 (associated with carrier strains). The nadA4 allele is located in the meningococcal chromosome like the other nadA alleles, and there are direct repeats before and after the gene insertion, putative promoter and terminator sequences, and a variable number of tetranucleotide repeats at the promoter level (7). All the nadA4-harboring strains were isolated in Norway during three different surveys spanning 8 years. These strains have different serotypes and serosubtypes, and from a genetic point of view, four of them are closely related. They belong to ST-26 and ST-1301, which differ by one allele only in the MLST scheme, whereas one strain (16260) belongs to a very different genetic environment (ST-37), which differs from ST-26 and ST-1301 in all seven MLST genes. NadA is not present in the 18 strains of the nonpathogenic human commensal N. lactamica examined. By analyzing the PCR patterns obtained with primers external to the nadA gene insertion site (7) of eight N. lactamica strains, we found that four of them gave the results typical of meningococcal strains lacking the nadA gene (that is, a 400-bp PCR product). Interestingly, the other four N. lactamica strains had in addition a 542-nucleotide insertion, a result different from those obtained for the other neisserial strains lacking the nadA gene. The insertion was sequenced and analyzed. There were no homologous sequences in the database. The insertion is flanked by 17-nucleotide direct repeats and is located 10 nucleotides upstream of the locus of insertion of the other nadA alleles (Fig. 1). The presence of this sequence in a subset of strains could be considered a relic of the early differentiation of two N. lactamica subgroups. This sequence is found only in this species.
nadA4 and its encoded peptide.The nadA4 gene codes for a protein (NadA4) consisting of 323 amino acids, which is shorter than the proteins encoded by alleles 1, 2, and 3 (NadA1, NadA2, and NadA3), which are composed of 362, 398, and 405 amino acids, respectively. An alignment of the four NadA alleles and the overall similar secondary structures of NadA4 and the other alleles are shown in Fig. 2. Secondary-structure analysis of NadA1, NadA2, and NadA3 revealed that each protein can be divided into three domains: an N-terminal globular head, an internal region with high coiled-coil propensity, and a C-terminal membrane anchoring region. Secondary-structure analysis of NadA4 revealed that the amino-terminal region (amino acids 24 to 90) exhibits 63% homology with the proteins encoded by the other alleles and has an undefined secondary structure. The internal region exhibits only 25% identity with other NadA forms, but surprisingly it maintains the α-helix propensity (60%). The carboxy-terminal region (amino acids 244 to 323), which spans the four amphipatic β-strands and is predicted to form the membrane anchor of the protein, is perfectly conserved (100%). Compared to other forms, NadA4 has a disrupted form of the leucine zipper motif.
To verify that the carrier strains were able to express NadA, total cell extracts and/or OMVs were loaded on an SDS gel and incubated with an anti-NadA4 mouse polyclonal antibody. All carrier strains carrying NadA1 or NadA2 were able to express detectable levels of the protein as high-molecular-mass oligomers, like the strains isolated from patients (data not shown). The results of the Western blot analysis of strain 65/96, bearing NadA4, are shown in Fig. 3. NadA4 was present in the total extract of strain 65/96 (Fig. 3, lane 2) and migrated at a molecular mass lower than that of NadA3 (Fig. 3, lane 1). The protein migrated at an apparent molecular mass of approximately 150 kDa and probably represented high-molecular-mass oligomers, most probably trimers (Fig. 3). As expected, NadA was not present in the NGH38 strain used as a negative control (Fig. 3, lane 9). The presence of NadA4 in the OMV fractions (strain 65/96) suggests that the protein is successfully exported to the outer membrane. Interestingly, NadA4 was recognized in Western blots by antisera raised against each of the three alleles, indicating that the differences in the sequences do not impair immune recognition (data not shown). To confirm that NadA4 forms stable high-molecular-weight oligomers, like the other forms, the OMV fractions were subjected to particularly severe denaturing conditions (Fig. 3, lanes 3 to 8). The remarkable stability in the presence of heat and reducing agents even in the absence of the leucine zipper motif (Fig. 2A) suggests that the coiled-coil region alone is able to promote stable oligomerization of protein monomers. The lack of cysteine residues in NadA4, like the lack of such residues in the other NadA forms, indicates that disulfide bonds are not involved in this phenomenon either. This result is consistent with results obtained for YadA (an adhesin/invasin of Yersinia species), which is homologous to NadA (12) and which is able to form stable high-molecular-mass aggregates, even if cysteine residues and a leucine zipper motif are not present.
NadA4 is able to induce antibodies with bactericidal activity and adhere to epithelial cells.To check whether NadA4 is able to induce antibodies with bactericidal activity, the protein, devoid of the leader peptide and of the anchoring domain (Fig. 2A), was expressed in E. coli as a His fusion protein, purified, and used to immunize mice in the presence of either Freund's adjuvant or alum as an adjuvant. Immune sera were then tested with a bactericidal assay by using strains 65/96 and 2996, which harbor NadA4 and NadA3, respectively. The results (Table 3) showed that NadA4 was able to induce antibodies which were bactericidal against the homologous strain when they were used in combination with either Freund's adjuvant or alum. However, the bactericidal activity against a NadA3-harboring strain was very low or absent. These results indicate that NadA4 is not able to mount a cross-bactericidal response, unlike NadA1, NadA2, and NadA3, which have been proven to induce bactericidal antibodies against each other, regardless of the sequence variations. Antibodies against NadA3 were highly bactericidal against the homologous strain (2996), whereas the bactericidal titer against strain 65/96, expressing NadA4, was low and present only when immunization was performed in the presence of aluminum hydroxide as the adjuvant. These results suggest that there are some protective epitopes which are common to NadA3 and NadA4, whose conformation could be affected by the adjuvant. NadA is able to bind epithelial cells. To verify that NadA4 has the binding activity of the other forms, recombinant NadA4 was used in a binding assay. Chang epithelial cells were incubated with increasing concentrations of recombinant NadA4 and with either recombinant NadA3 as a positive control or an unrelated protein, recombinant protein NMB2132 (18), as a negative control. Binding was detected by fluorescence-activated cell sorting. As shown in Fig. 4, NadA4 binding was dose dependent, and the mean fluorescence index was approximately 30% reduced compared to that of NadA3, probably indicating reduced binding affinity.
A major unanswered question about N. meningitidis is the degree of similarity between the strains isolated from healthy individuals (carrier strains) and the strains isolated from people with disease. Apart from the capsule (6), which can account for the overall higher rate of nongroupable strains in carriage strains than in pathogens, no other pathogenicity factors could consistently explain the difference between strains able to evoke disease and commensals. In this work, we found that the nadA gene is an additional genetic marker that segregates differently in the two populations; while it is present in 50% of the strains isolated from patients and in members of three of the four hypervirulent clusters, it is largely absent in most of the strains isolated from healthy people (only 16.2% of such strains carry the gene). Remarkably, in this population, most of the nadA-positive strains (18 of 25 strains) belonged to the ET-5 and ET-37 hypervirulent lineages. Strains belonging to hypervirulent clusters are seldom isolated from healthy individuals (less than 10%) (5) and are apparently indistinguishable from homologous strains isolated from patients. The ET-5 and ET-37 strains used in this study represent the overlap between the two populations. The differences between hypervirulent cluster strains isolated from patients and hypervirulent cluster strains isolated from healthy individuals should be analyzed further in order to understand whether the different behaviors of genetically similar, if not identical, strains could be due to different abilities of the isolates to cause disease or due to different susceptibilities to disease of the different hosts. When we examined the carrier strains used in the present work that did not belong to any hypervirulent cluster, we found that the nadA gene was present in only 5.1% of them. Interestingly, all but two of these strains harbored a new nadA allele which we found only in carrier strains and which we designated nadA4. The NadA4 protein is expressed and found in the bacterial surface, and it is able to induce antibodies with bactericidal activity and to bind to epithelial cells like the other three forms of NadA. Questions to be answered in the future are whether we identified, in the carrier strain population, a subgroup of isolates which are potentially more virulent than the isolates lacking NadA and, if NadA4 does not confer any additional virulence properties, what the function of this protein is. In conclusion, we believe that a partial answer to the question of why in most countries only a few individuals colonized by N. meningitidis develop disease is beginning to emerge. Most of the people are colonized by strains which lack factors that are essential for virulence, such as the capsule, or that are potentially important for virulence, like NadA. These strains are unlikely to be able to cause disease and should be considered commensals, like N. lactamica, which also lacks the capsule and which, as shown in this work, also lacks the nadA gene. However, the commensal strains are likely to play an important role in providing exchange of genetic material and in priming immunity against the virulent strains (9, 15, 16, 21).
We thank C. Mallia for editing the manuscript, G. Corsi for providing artwork, R. Rossi for providing the OMV preparation, and R. Moxon for providing eight N. lactamica strains. In this work we used the MLST website (http://www.mlst.net ) developed by Man-Suen Chan and Keith Jolley; the development of this site is funded by the Wellcome Trust.
- Received 10 December 2003.
- Returned for modification 15 January 2004.
- Accepted 11 February 2004.
- Copyright © 2004 American Society for Microbiology


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Can Meditating Help With Weight Loss?
Is it possible to lose weight by meditating?
Yes, it actually is.
As crazy as it may sound, meditation has been proven to be a great complement to diet and exercise.
As more people start to appreciate the power of the mind body connection, more dieters will discover just how effective mindful eating is.
Here's how meditation can help with weight loss…
Meditation Relieves Anxiety
When it comes to weight loss emotional eating is one of the biggest obstacles dieters run into.
Emotional eating is usually a result of stress and anxiety.
This can be brought on for a variety of different reasons. But whatever the reason, the sooner you deal with it the better.
Meditation is a great way to relieve stress as it helps put you in a state of peace.
As you continue to find better ways to deal with your emotions you will start to find emotional eating is a thing of the past.
Meditation Can Give Your Self Esteem a Boost
Most people don't realize it but your level of self esteem plays a huge role in whether or not you reach your weight loss goals.
When you don't truly love yourself you will find it very difficult to shed unwanted pounds.
On the other hand, you will find that weight loss is a lot easier when you feel good about yourself.
Becoming less judgmental of yourself is the starting point to becoming healthier and happier.
Meditation Can Give Your Motivation a Boost
Weight loss can already be a difficult task in and of itself.
When you lack motivation that task becomes even more difficult.
The great thing about meditation is it helps you build a strong mental foundation.
Having a strong mental foundation will provide you with the motivation you need to resist the temptation to grab another bag of chips.
Meditation Can Help You Develop a Healthy Relationship With Food
Most people look at food as their enemy.
This is usually because they have been fed a bunch of lies about what they can and can't eat while on their weight loss journey.
First and foremost food is not your enemy. Or at least it doesn't have to be.
Once you start to change your attitude about food you will find that weight loss becomes easier.
Top 5 Meditation Practices For Those Looking To Lose Weight
Always Listen To Your Body
These days it is extremely easy to become disconnected from your own body.
This is in large part due to how we live our lives.
Many of us were taught at an early age to clean our plates.
Even if we were full we had to continue eating until the plate was empty.
As a result we started to ignore our bodies and continue to eat even when we aren't hungry.
Meditation helps you become more sensitive to what your body is telling you.
By listening to your body you will be less likely to overeat.
Become a More Mindful Eater
You want to know one of the best weight loss tactics?
Its learning how to become a more mindful eater. During meditation you become more mindful.
In most cases this will flow over into other areas of your life.
The idea is to learn how to savor every bite of food you eat.
Learn To Talk Positive About Yourself
During your meditation sessions you should focus only on the things you love about yourself.
Focus on the positive efforts you have been making to reach your weight loss goals.
By focusing on the things you love about yourself you will start to enjoy life more.
You will also be less likely to look at cutting back on calories as a bad thing.
Set Clear Goals
Your meditation session is the perfect opportunity to set clear goals.
How much weight do you want to lose? In what time frame do you want to reach that goal?
This is also a great time to evaluate your progress.
How close are you to reaching your goals and what changes can you make to improve things?
Analyze Your Setbacks
Every weight loss journey is bound to have a few setbacks.
If you take the time to find what causes you to get off track you can make plans to deal with it.
For example, if you know you always overindulge on Thanksgiving, prepare in advance so you won't trip up.
Instead of going to the Thanksgiving party hungry, have a small meal a few hours before you go.
Doing so can prevent you from overeating as you won't be as hungry.
You should also have a glass of water right before you eat.
Water will fill you up and make it less likely you will overindulge.
If you need to, find something completely different to do instead of subjecting yourself to all that delicious looking food.

subset: commoncrawl


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The Public Eye: Fool Me Once, Fool Me Twice
A familiar American aphorism is "Fool me once, shame on you; fool me twice, shame on me." In 2000, Karl Rove convinced Americans that a relatively unknown Texas governor could be a competent president. In 2008, John McCain's Rove, Steve Schmidt, argues that an even more obscure Alaska governor will make a credible vice president and likely 45th president. Are Americans about to be fooled again?
In 2000, Rove ran a Republican presidential campaign based upon personality rather than issues. His candidate, Texas governor George Bush, was affable, able to deliver a simple speech with conviction, and a born-again Christian. Even though he was the son of a former president and had a long connection with national politics, Bush was packaged as an outsider: someone who could come to Washington, reach across party lines, and get America back on track.
In 2008, Schmidt also runs a Republican campaign based upon personality. Recently a McCain spokesman acknowledged: "This election is not about issues. This election is about a composite view of what people take away from these candidates." Alaska Gov. Sarah Palin is affable, able to deliver a simple speech with conviction, and a born-again Christian. Even though she's running with John McCain, who's been in Washington for 26 years, McCain-Palin are being packaged as outsiders, "mavericks"—people who can come to Washington, reach across party lines, and get America back on track.
The Rove-Schmidt strategy is targeted primarily at hard-core, born-again Christians—somewhere between one-quarter and one-third of the U.S. electorate and a key component of the Republican base. Although in the past three years he's learned to walk and talk like a born-again, John McCain hasn't convinced the Republican faithful that he is a true believer. The selection of Sarah Palin as his running mate is designed to win them over.
In 2000, George Bush came out as a born-again Christian. In 2008, Sarah Palin also advertises her identity as a born-again. Bush and Palin believe the Bible is literally true and God intervenes in worldly affairs. Bush said, "God told me to invade Iraq." Palin says the Iraq war is a "task that is from God." Both believe life begins at conception and abortion is a mortal sin; therefore, Bush and Palin insist that the federal government intervene in what most Americans regard as a private family medical decision. Both believe homosexuality is "a lifestyle choice"—Palin believes homosexuality can be "cured" through prayer. In 2000, Bush said there was insufficient evidence that global warming is man made; in 2008, Palin agrees. Both argue that creationism should be taught in the schools.
In 2000, many claims that George Bush made about his bona fides were later found to be false. He said he had been a successful CEO, but his businesses were failures. Bush said he had been a pro-environment governor, but Texas was notorious for unregulated pollution. In 2008, many claims that Sarah Palin has made about her credentials are also false. She says she was a successful mayor, but she actually left her town deeply in debt. She claims to be against "pork barrel" projects, but she lobbied for them as mayor and governor, most famously the "bridge to nowhere."
Eight years after the great hoax of 2000, many political observers fear that Sarah Palin's celebrity-buzz will propel the Republican ticket to victory and saddle the United States with four more years of failed Republican policies. Democrats want to know what the Obama campaign should do to counter this onslaught. First they need to understand what Sarah Palin represents.
University of California Linguistics Professor George Lakoff explains the nature of Palin's appeal: She's "the mom in the strict father family, upholding conservative values. Palin is tough ... She is disciplined: raising five kids with a major career. She lives her values... She has the image of the ideal conservative mom: pretty, perky, feminine, Bible-toting, and fitting into the ideal conservative family."
Lakoff argues that to combat this evocative symbol, a GOP campaign based upon personality rather than upon substance, the Obama campaign needs to attack McCain and Palin on their values and symbolism: "Democrats ... need to call an extremist an extremist: to shine a light on the shared anti-democratic ideology of McCain and Palin, the same ideology shared by Bush and Cheney. They share values antithetical to our democracy."
But this won't be sufficient. If Americans are to avoid being fooled again, the Obama campaign should attack on three fronts between now and Nov. 4: First, they must keep talking about the economy and tie McCain-Palin to failed Bush policies that have produced an economic recession. Second, they should use the L word; repeatedly point out that McCain-Palin have lied about their records and key elements of their political personas—emphasize they are not "mavericks," but rather "scoundrels." Finally, Dems must seize the moral high ground: Emphasize key American values such as equity, empowerment, and personal responsibility and make it clear that McCain-Palin's values are un-American.
Bob Burnett is a Berkeley writer. He can be reached at email@example.com..


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We have included all charges provided to us by the property. However, charges can vary, for example, based on length of stay or the room you book.
Le 61 Berthier Hotel Area
Distances are calculated in a straight line from the property's location to the point of interest or attraction, and may not reflect actual travel distance.
Distances are displayed to the nearest 0.1 mile and kilometer.
Espace Champerret - 0.6 km / 0.4 mi
Parc Monceau - 1.1 km / 0.7 mi
Palais des Congres de Paris - 1.3 km / 0.8 mi
Square des Batignolles - 1.4 km / 0.9 mi
Place Charles de Gaulle Etoile - 1.4 km / 0.9 mi
Arc de Triomphe - 1.4 km / 0.9 mi
Musee Nissim de Camondo - 1.5 km / 0.9 mi
Parc Clichy-Batignolles - Martin-Luther-King - 1.5 km / 1 mi
Lido - 1.6 km / 1 mi
Jacquemart-Andre Museum - 1.6 km / 1 mi
Bois de Boulogne - 1.7 km / 1 mi
Baccarat Museum - 2.1 km / 1.3 mi
Place de Clichy - 2.2 km / 1.4 mi
Elysee Palace - 2.3 km / 1.4 mi
Jardin d'Acclimatation - 2.3 km / 1.4 mi
The nearest major airport is Paris (ORY-Orly) - 18.3 km / 11.4 mi.
Le 61 Berthier Hotel Amenities
- wifi wifi-free
- local_cafe tea/coffee making facilitiesbell boy service
Le 61 Berthier Hotel Taxes & Fees
Please note that a compulsory city tax is required to be paid by the GUESTS on top of B&A's contracted rates on all booked hotel accommodation in Paris, France. The fees are as follows: - € 4.40 PER PERSON PER NIGHT for stays in 5+ star hotels - € 3.30 PER PERSON PER NIGHT for stays in 5 star hotels - € 2.48 PER PERSON PER NIGHT for stays in 4 star hotels - € 1.65 PER PERSON PER NIGHT for stays in 3 star hotels - € 0.99 PER PERSON PER NIGHT for stays in 2 star hotels - € 0.83 PER PERSON PER NIGHT for stays in 1 star hotels Please note: This tax is to be paid by the GUESTS directly at the hotel before the end of the stay.


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Mermaid tails for all mermaids and mermen in the UK.
These mermaid fins make your dream come true! Have you always dreamed of becoming a real life mermaid? Then you must have one of these beautiful mermaid tail costumes for swimming. Our tails look realistic and are professional equipment especially developed for mermaiding and freediving.
All our mermaid tails include a professional freediving mono fin. The mermaid mono fin is made out of polypropylene. It consists of two separate foot pockets which are made out of a soft rubber. The two heel straps are adjustable and guarantee the perfect fit for your feet. Our mono fins don't consist of any screws or nails which could rust and damage your mermaid tail.
These mermaid tails will turn you into a mermaid princess
This swimmable mermaid tail costume consists of swimwear fabric with 85% polyester and 15% spandex. A scale print gives your new mermaid tail the perfect look.
Please note: Mermaid tails are NO BUOYANCY AIDS! Mermaid tails are FOR EXPERIENCED SWIMMERS ONLY. The use of mermaid fins in the water should consequently not be used without supervision or an experienced buddy. Additionally we recommend a minimum age of 8 years.


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Liturgical Colour : Green or White (Saturday Mass of Our Lady)
Dear brothers and sisters in Christ, today as we listened to the words of the Scripture we are all reminded of the missions which God has entrusted to us as the people of God, whom He has called to be His servants and followers. We heard first of all the story of the calling and anointing of Saul as the first king of Israel by the prophet Samuel, and then the story from the Gospel on how the Lord Jesus called Levi the tax collector to be one of His disciples.
Therefore today we heard the story of the calling of two men by God to be the instruments of His works among His people. Saul was called from the tribe of Benjamin, the smallest and least important of the tribes of Israel at that time; and he was also from the least important and smallest of the groups and families within the tribe of Benjamin. And God called him to be the first king over His people Israel when the people cried out to have a king over them.
Saul was just a simple man without prestigious or powerful background, and God chose him to be the king over His people, much as how later on He would also choose a simple shepherd boy, David, the youngest and smallest of the many sons of Jesse to be the successor of Saul. God indeed chose His servants and called them, and not the other way round, that is we choose ourselves. He made those whom He had chosen to be worthy and empowered them as how He led Saul and David to many victories over their enemies.
In our Gospel passage today, we then heard of the calling of the tax collector Levi, whom the Lord Jesus called to be His disciple. Levi left behind his job and everything he had, following the Lord and eventually would be known by a new name, that is Matthew. As one of the Twelve Apostles, he would be known as St. Matthew, the Apostle and Evangelist as not only that he was one of the Twelve but with St. John the Apostle he also had the distinction of being also the writer of one of the four Holy Gospels.
When Levi invited the Lord to have a meal with him and the other tax collectors at his place, the Pharisees and the teachers of the Law immediately judged Him and those tax collectors, condemning His actions and questioning why a supposedly holy and popular Man of God would mingle with the tax collectors who were considered among the least welcomed and also hated by a large portion of the population.
The tax collectors were despised because they were seen as collaborators who 'sold' their own countrymen and people to the Romans as they helped to collect the hated taxes and money for the Roman governors and magistrates. But in truth, those tax collectors were just the same as anyone else, when looked upon without the bias that many were looking at them with. In fact as we can see, while the tax collectors welcomed the Lord and were willing to listen to Him, it was the Pharisees and the teachers of the Law who refused to believe in the Lord and rejected Him.
Again, as I said earlier on, God chose and called those whom He wished to be His followers. And unlike many of the Pharisees and teachers of the Law who wanted to show off their piety and righteousness before the people, and wanted to justify themselves as being better than others especially the tax collectors, they actually failed to see that God honours the humble and those who love Him more than they love themselves.
Now, brothers and sisters in Christ, how is this significant for us? It is so because we are all also called by God to be His disciples. God has called us to follow Him through our baptism, and if we are truly willing to follow Him, then we should open our hearts and minds to welcome Him into our midst. Are we able to do this? Are we able to trust in God to lead us down the right path going forward? God will lead us down the right path if only we allow Him to guide us.
Let us all seek the Lord and allow Him to guide us in our journey of life and faith. Let us dedicate ourselves anew to Him, and let us grow in our trust and faith with each and every days of our lives from now on. May God bless us all and may He be with us always, through all the goods and trials of this life we have in this world, that we may be His faithful disciples and witnesses to the nations. Amen.


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Fiesta committee to distribute free tickets to concert, rodeo
September 14, 2004
Some of the people sitting along the Greeley Fiesta parade route Saturday morning will get a little more than a view of fancy cars and festive musicians.
They'll get free tickets to other weekend Fiesta events.
Members of the Fiesta committee will distribute 50 tickets to Saturday's Kumbia Kings concert and 40 tickets to Sunday's Mexican Rodeo/Charreada during the parade. The distribution will be done randomly and secretly, organizers said.
The parade will begin at 10 a.m. Saturday at Greeley Central High School, 15th Avenue and 14th Street. The route will go north on 14th Avenue to Island Grove Regional Park, 14th Avenue and A Street.
Doors for the concert, which features AB Quintanilla III y los Kumbia Kings, opens at 8 p.m. Saturday at the Community Building at Island Grove. Tickets cost $25 and will be sold in advance at the park on Saturday. Tickets at the door will cost $30.
The Mexican rodeo begins at 4:30 p.m. Sunday at Island Grove.


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Normally, SVWM does not advertise products or services but we have made an exception with this because it is an important addition to the knowledge base of WWII events and worthy of mention.
For several years, Jan Nieuwenhuis has been compiling information regarding allied plane crashes in the Netherlands during WWII. He has amassed a great deal of information. Not only has he gathered all this information in one place, Nieuwenhuis also created a computer application that you can use to search the data he has compiled and continues to add to. Jan Nieuwenhuis is a member of the board of the Aeronautical and War Museum on Texel Island.
His AirWarWWII program is free to download and use. It connects to the web database that he continues to augment on a regular basis. It contains information on more than 1000 crashes and over 5000 crew members. The screenshots below will give you an idea of what it looks like. Click an image to see a larger view. If you have friends and/or family or are curious about air crashes, aircraft and their crews you need to take a look at this.
Main Screen
Aircraft Search
Crash Search
Crew Search
Cemetery Search
Search Results
Crash Details


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New insights into the antifungal activity of Taxus baccata L.
Taxus baccata L., yew, is dioecious, small to medium-sized evergreen tree, native to Europe, Africa and Asia, but it is cultivated worldwide because of its ornamental features. It is long known that all parts of yew (except for aril) are toxic, due to the alkaloid taxine. Nevertheless, some parts of yew tree can be used to treat cancer and as antimicrobial agents. Unlike its antibacterial properties, antifungal activity of T. baccata is poorly investigated. In this research, antifungal activity of yew methanolic and aqueous extracts was tested against Candida albicans ATCC 10231, Aspergillus brasiliensis ATCC 16404, and Ascosphaera apis MUCL 30769, through the agar well diffusion method. Leaves, bark and reproductive structures extracts were prepared separately for male and female plants. Unlike the aqueous, methanolic extracts caused variable degree of fungal growth inhibition. The strongest inhibition was observed in the activity of the aril extract against A. brasiliensis and C. albicans, with the activity of the male bark extract against A. apis following close. Considering the emerging multidrug resistance in C. albicans, an interesting finding is the inhibition of this species by all tested methanolic extracts, which is significantly stronger in comparison to the inhibition by standard antimycotic solution. According to the available data, male reproductive structures of this species were not tested earlier for their antifungal activity, and our study showed high level of antifungal activity of methanolic microstrobili extract.
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
©Genetics & Applications, The Official Publication of the Institute for Genetic Engineering and Biotechnology, University of Sarajevo


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- Amnesty International USA
- Amnesty International USA Turkey Country Page
- Andrew Finkel's Latitute Blogs
- BIA net English Language Page
- Cem's International Turkish Digest
- GIT-North America
- Human Rights Now
- Human Rights Watch
- Jenny White's Kamil Pasha
- Journalist in Turkey
- POMED: Project on Middle East Democracy
Amnesty USA, Turkey Regional Action Network
Tag Archives: DirenGezi
Amnesty calls on Turkey: "Respect right to peaceful assembly" on May 1
In advance of annual May 1 demonstrations, Prime Minister Tayyip Erdogan is talking tough again. "Give up your hope of Taksim," he said. According to Turkish media, tens of thousands of police have been mobilized. In a public statement on … Continue reading
"Twitter Trial" in Izmir
Coming in the context of repeated efforts by the Turkish government to block social media outlets, the next hearing of the Izmir "Twitter Trial" is receiving international attention. The case involves 29 men and women on trial Twitter messages they … Continue reading
Posted in Amnesty International, Freedom of Assembly, Freedom of Expression, Gezi Park, Internet Freedom, Taksim, Uncategorized Tagged Amnesty International, Andrew Gardner, Censorship, criminalization of dissent, Diren Gezi, DirenGezi, Erdogan, Freedom of Association, Gezi Park, Human Rights, internet freedom, Izmir Twitter trial, taksim square, Turkey, YouTube
The frustrating road to justice in the Sarısülük case
The case of Ethem Sarısülük, killed by a policeman's bullet during the Gezi protests, has been one of on-going concern for Amnesty International. Yesterday, the trial had its fourth, inconclusive hearing and Amnesty was there to follow the case.
A late night attack on internet freedom
Turkish Prime Minister Tayyip Erdogan was as good as his word. According to Reuters, he told a friendly crowd at a rally in Bursa earlier today: "Twitter, mwitter!," Erdogan told thousands of supporters at a rally in the northwestern province … Continue reading
Ali Ismail would have been Twenty today
Ali Ismail Korkmaz would have been twenty today. Instead, he was brutally beaten to death last June during the Gezi protests. There is substantial evidence of police involvement in his murder and in a police cover up after the fact. … Continue reading
The Trail of Impunity: Roboski and Okmeydani
The protests which have shaken Turkey in the aftermath of Berkin Elvan's death at age fifteen have generated many shocking images. Grainy photographs from protests in Roboski/Qileban, however, struck a particular nerve with me because they highlight the extent to … Continue reading
Posted in Amnesty International, Excessive Force, Kurds, Turkey Tagged Amnesty, Andrew Gardner, Berkin Elvan, DirenGezi, Erdogan, excessive force, Gezi Park, Human Rights, impunity, Kurds, Qileban, Roboski, taksim square, tear gas, Turkey, Uludere
Neither apology nor call for justice
Many observers wondered how Turkish Prime Minister Tayyip Erdogan would respond to the death of Berkin Elvan. Berkin Elvan was a fourteen year old boy who was struck in the head by a policeman's tear gas canister last June. He … Continue reading
Amnesty issues statement on human rights and urban transformation projects in Turkey
In advance of the March 30 local elections in Turkey, Amnesty International has issued a statement calling on all parties to commit to "[conducting] urban transformation projects only in a way which respects the human rights of all residents." The … Continue reading
AI Turkey Researcher Andrew Gardner blogs from Istanbul
Amnesty International's researcher on Turkey has been in Istanbul monitoring events. This morning, Amnesty his blog. He writes: All the evidence suggests that Berkin died because of injuries he sustained last 16 June, when he was hit at close range … Continue reading
Posted in Uncategorized Tagged #BerkinElvan, #BerkinElvanÖlümsüzdür, Amnesty International, Andrew Gardner, Berkin Elvan, Diren Gezi, DirenGezi, Erdogan, excessive force, Freedom of Assembly, Freedom of Expression, gas canisters, Gezi Park, impunity, Istanbul, Police violence, taksim square, tear gas, Turkey, water cannon
Tensions as Berkin Elvan is laid to rest
Yesterday, we carried news of fifteen year old Berkin Elvan's death. He weighed less than 36 pounds at the time of his death. In Turkey and in cities throughout the world protests have been held in mourning and in anger. … Continue reading
Posted in Amnesty International, Excessive Force, Freedom of Assembly, Freedom of Association, Freedom of Expression, Turkey Tagged #BerkinElvan, Abdullah Gul, Amnesty, Amnesty International, Andrew Gardner, Berkin Elvan, Diren Gezi, DirenBerkin, DirenGezi, excessive force, Freedom of Assembly, Freedom of Expression, gas canisters, Gezi Park, Human Rights, impunity, Istanbul, protests, taksim square, tear gas, water cannon


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Car Hire
Ellis Exclusive Villas are happy to recommend two Orlando car hire companies. Car on the Drive offers a wonderful pick-up from the airport service. For more seasoned Orlando travellers, RentalCars.com offers a more traditional but equally great-value choice.
Car on the Drive
Car on the Drive is a family-owned and run business based in Orlando.
It's run by Peter and Wendy who really go the extra mile to keep their customers happy.
Car on the Drive will pick you up from the airport and take you back to your villa – leaving your car on the drive for you. This is a huge benefit when you've just stepped off a long trans-Atlantic flight with kids in tow: no need to negotiate the busy traffic and road system around Orlando airport. No need to queue for car hire, or find the car hire office.
- No more waiting in a queue
- No struggling with luggage
- FREE meet & greet pick up service by an experienced local driver
- No more getting lost or map reading on the way to your villa
- FREE pick up from your villa back to the airport.
Some of our guests have even enjoyed some impromptu Orlando driving lessons from the Car on the Drive staff after being dropped off at their villa! This is what we mean by the extra mile: if you aren't comfortable with driving on the right or using an automatic, the car on the drive team are so helpful. They'll also take you back to the airport when it's time to leave.
It's a great service and one we highly recommend to all our guests.
If you prefer a more traditional hire car agreement and prefer to make your own way to your vacation property, Ellis Exclusive Villas are pleased to also recommend RentalCars.com
Rental Cars have many locations in and around the Orlando area, including Orlando International Airport. The company doesn't make surcharges for credit card payments and it is quick and easy to make or amend bookings online.


text: ESC site newshttp://esc-duesseldorf.de/news/2020-08-06T11:55:53ZUpdates on events by the English Speakers' Circle.Games evening – English Scrabblehttp://esc-duesseldorf.de/news/event/189/13 Jun 2020: It's eleven years since we had an evening entirely devoted to this game, useful preparation for wet holiday evenings, perhaps!Long live the short story!http://esc-duesseldorf.de/news/event/188/9 May 2020: The Orange Planet Theatre will perform dramatized readings of five of the best-loved classic short stories ever written: Anton Chekhov's "Oh! The Public", Edgar Allan Poe's "The Tell-Tale Heart", Guy de Maupassant's "The Necklace", Katherine Mansfield's "The Singing Lesson" and W. W. Jacobs' "The Monkey's Paw".A short trip through the life of a textile designerhttp://esc-duesseldorf.de/news/event/187/25 Apr 2020: Susann Sieben will tell about the challenges and changes from her time working as a designer for the German textile industry, also showing us examples of her own designs.Oliverhttp://esc-duesseldorf.de/news/event/191/by Peter Griffith, based on the novel by Charles Dickens. Performed by White Horse Theatre. 27 Mar 2020: Performed here very successfully ten years ago, and using Dickens' original language, this play captures the essence of the novel "Oliver Twist", which is a passionate plea for justice for the poor and a heart-warming tale of hope in the midst of adversity.Dickens' romantic heroineshttp://esc-duesseldorf.de/news/event/186/14 Mar 2020: Britta Zangen illustrates how, in that very Victorian age, when the emancipation of women was being openly discussed and when industrialisation was changing human life radically, Dickens presents almost all the young heroines of his 15 novels as soft, kind, pure and beautiful.


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The UberCloud Compendium of Case Studies is Coming in December !
Now in its 4th annual edition, this 2016 UberCloud Compendium of 18 engineering cloud case studies will again be published in December, demonstrating another milestone towards the democratization of High Performance Computing (HPC) applied to engineering and scientific application simulations. HPC is still widely recognized as a complex computing tool which is mastered by a relatively small and eluded crowd of supercomputing experts who apply HPC to sophisticated fields of applied mathematics, physics, chemistry, life sciences, energy, weather and climate, and more. Supported by UberCloud's novel software container technology, the various teams have tackled complex tasks, ranging from acoustic modelling and simulating dry-type transformers to air flow through an engine intake and the simulation of wind turbines, to thermal modelling of a reactor. HPE and Intel are involved in these UberCloud HPC Experiments again as Sponsors. Both have a strong and long-term HPC private and public cloud strategy, with the goal of helping hundreds of thousands of enterprises take full advantage of the benefits of HPC, including those stemming from the UberCloud HPC Experiments. Watch out for the announcement of the 2016 UberCloud Compendium !


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- 220Shares
Let's take a look at how to organise a drawer in your hallway today. It's a quick weekend project that will take no time at all, but is a really great win for the space. Let's get started on exactly how to get it sorted, shall we….
Last weekend I did a quick project in my Hallway that has been on my TO DO list for a while, and I totally LOVE the results.
As such, I wanted to share it with you today, in the hope that you get inspired to try something like this yourself.
Now – this is how I organised my Hallway drawer – BUT you can use the tips to organise pretty much any drawer in your house.
I, of course, used the A.D.O.R.E method to get it done (Assess, Declutter, Organise, Real Life, Evaluate), so I'll take you through it step by step – hope you enjoy!
Of course, the very first step of any organising project is to Assess / Plan it out.
For this drawer, it's the last storage space before we leave the house, and the first as we enter – so it was the perfect place to put all those little things that we need to grab before we go out (and conversely, to drop down when we get back).
The Hallway Drawer is the perfect place for things like the following: –
- Keys – car keys / house keys etc…
- Letter Writing Essentials (Envelopes, Stamps, Pen) – so you can quickly fill out forms and send them back without having to search high and low for what you need
- Sunglasses – how many times are you searching for these when you need them!
- Spare Change – great idea for when you need some change, or for men especially to empty their change from their trousers when they walk in
- Comb / Brush – last minute straightening before you leave the door
- Lipstick / Lip Balm – again for last minute checks in the mirror
Now I knew what we DID want in the drawer (the day to day stuff we needed to grab and have easy access to), it was time to look and see what was actually in there: –
Now – we had much more than this list inside the drawer, as you can see it had got out of hand a little!
The extras that really weren't needed right there were: –
- Cheque Book – better in our command centre / paperwork area
- Maps / Walking Route books etc… – Better in our holiday area as they aren't used on a daily basis
We also had lots of little bits and bobs that we had all just thrown in there over time – yuck – hate this stuff so much. It always seems to grow more when the space is already disorganised because it can hide a little more – another great reason to get it sorted!
It took less than 5 minutes to go through the drawer and declutter what wasn't needed.
I moved everything that was in the wrong place to the correct place – and binned the rubbish (recycling where possible).
Already it felt better!
The next step of the ADORE Method is O – Organise.
Now I had everything I wanted to put inside the drawer, it was time to work out the best way to organise it so it was: –
- Easy to grab things
- Easy to see what's in there
- Easy to put things away
As with most drawer, having compartments works best – so you can have a section for each different thing.
So – I measured the space available and went searching online for the perfect sized storage.
It takes a while sometimes to find the exact right item, but it's worth waiting, because then you get the right thing.
… and I found the perfect size for what I wanted – with the right number of compartments as well** – win!
STEPs 4 & 5 – LIVE WITH IT (REAL LIFE) AND TWEAK IF NECESSARY (EVALUATE)
Of course, as this is a new project, we are going to have to live with it for a while to see whether it's right or not.
You can't tell completely unless you use it – so I'll give it at least a month to see how it goes.
The proof will be if we look at it in a month and it's looking pretty much the same as it is now.
If not – then I know that I will need to tweak things so that they work even better. Maybe we'll find that we need to store other items there, or that we don't need everything in it that I think we do.
Who knows!
But for now – we're all really pleased with the finished result…
And there you have it – How to organise a hallway drawer – step by step
I hope you've got some inspiration for doing this in your own hallway, it really is a quick win that can make a LOT of difference to your every day life, so it's a great project to get started with.
If you follow the ADORE Method, then you'll find it so easy to do, and if you want a refresh of the entire Method, then just click here for more details.


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Spring Moon - The Peninsula Hong Kong
Top Tags:
It was a birthday celebration and the dinner was superb! Excellent service and food, ambience was perfect too. Highly recommended. The 91 year-old birthday girl was happy!
Friends took us to Spring Moon during our five-day stay in Hong Kong. The food was so exquisite, and the atmosphere so delightful, that we went back again two days later. Best and most interesting dim sum we have ever had.
Duck lunch, beautifully prepared, delicious. Fantastic tea. And genuinely atmospheric.
A classic restaurant which stays true to it's origins. Not the place to come if you want something more creative or modern. The food was fine, the service attentive. Main issue was the amount of noise from other diners which were mainly families. Lot's of people using laptops at the table. Not really like any other Michelin star restaurant I have visited, but I guess that's what you get when the restaurant is located in a big hotel.
Good place and good foods especially dimsum
The best dim sum I've ever had (and I'm chinese). The BBQ pork puff was divine. Will definitely return again to HK just to have their dim sum.
Very upscale superb meal! Would strongly recommend the Peking Duck which is the best I've ever had!
Delicious! We had a nice and elegant dinner, the service is excellent and the food is delicious!
Awesome! Derack - the Manager - is fantastic!
Delicious! It's a most in Hong Kong! We had a nice and elegant dinner, the service is excellent and the food is delicious!
Spring moon's dim sum is very delicious but the pork is too salty,maybe they can change.
Spring Moon - The Peninsula Hong Kong is rated 4.8 stars by 12 OpenTable diners.
You can book this restaurant by choosing the date, time and party size.


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|The global methane budget 2000-2012
|Publication Type
|Journal Article
|Year of Publication
|Saunois M., Bousquet P., Poulter B., Peregon A., Ciais P., Canadell J.G, Dlugokencky E.J, Etiope G., Bastviken D., Houweling S., Janssens-Maenhout G., Tubiello F.N, Castaldi S., Jackson R.B, Alexe M., Arora V.K, Beerling D.J, Bergamaschi P., Blake D.R, Brailsford G., Brovkin V., Bruhwiler L., Crevoisier C., Crill P., Covey K., Curry C., Frankenberg C., Gedney N., Hoglund-Isaksson L., Ishizawa M., Ito A., Joos F., Kim H.S, Kleinen T., Krummel P., Lamarque JF, Langenfelds R., Locatelli R., Machida T., Maksyutov S., McDonald K.C, Marshall J, Melton J.R, Morino I., Naik V., O'Doherty S, Parmentier F.JW, Patra P.K, Peng C.H, Peng S.S, Peters G.P, Pison I., Prigent C., Prinn R., Ramonet M., Riley W.J, Saito M., Santini M., Schroeder R., Simpson I.J, Spahni R., Steele P., Takizawa A., Thornton B.F, Tian H.Q, Tohjima Y., Viovy N., Voulgarakis A., van Weele M., van der Werf G.R, Weiss R., Wiedinmyer C., Wilton D.J, Wiltshire A., Worthy D., Wunch D., Xu X.Y, Yoshida Y., Zhang B., Zhang Z., Zhu Q.
|Earth System Science Data
|Date Published
|Type of Article
|ISBN Number
|Accession Number
|atmospheric; biomass burning emissions; composition; greenhouse-gas emissions; hydroxyl radicals; intercomparison project accmip; isotopic; natural-gas; past 2 decades; process-based model; trace gases; tropospheric methane
The global methane (CH4) budget is becoming an increasingly important component for managing realistic pathways to mitigate climate change. This relevance, due to a shorter atmospheric lifetime and a stronger warming potential than carbon dioxide, is challenged by the still unexplained changes of atmospheric CH4 over the past decade. Emissions and concentrations of CH4 are continuing to increase, making CH4 the second most important human-induced greenhouse gas after carbon dioxide. Two major difficulties in reducing uncertainties come from the large variety of diffusive CH4 sources that overlap geographically, and from the destruction of CH4 by the very short-lived hydroxyl radical (OH). To address these difficulties, we have established a consortium of multi-disciplinary scientists under the umbrella of the Global Carbon Project to synthesize and stimulate research on the methane cycle, and producing regular (similar to biennial) updates of the global methane budget. This consortium includes atmospheric physicists and chemists, biogeochemists of surface and marine emissions, and socio-economists who study anthropogenic emissions. Following Kirschke et al. (2013), we propose here the first version of a living review paper that integrates results of top-down studies (exploiting atmospheric observations within an atmospheric inverse-modelling framework) and bottom-up models, inventories and data-driven approaches (including process-based models for estimating land surface emissions and atmospheric chemistry, and inventories for anthropogenic emissions, data-driven extrapolations). For the 2003-2012 decade, global methane emissions are estimated by top-down inversions at 558 TgCH(4) yr(-1), range 540-568. About 60% of global emissions are anthropogenic (range 50-65 %). Since 2010, the bottom-up global emission inventories have been closer to methane emissions in the most carbon-intensive Representative Concentrations Pathway (RCP8.5) and higher than all other RCP scenarios. Bottom-up approaches suggest larger global emissions (736 TgCH(4) yr(-1), range 596-884) mostly because of larger natural emissions from individual sources such as inland waters, natural wetlands and geological sources. Considering the atmospheric constraints on the top-down budget, it is likely that some of the individual emissions reported by the bottom-up approaches are overestimated, leading to too large global emissions. Latitudinal data from top-down emissions indicate a predominance of tropical emissions (similar to 64% of the global budget, <30 degrees N) as compared to mid (similar to 32 %, 30-60 degrees N) and high northern latitudes (similar to 4 %, 60-90 degrees N). Top-down inversions consistently infer lower emissions in China (similar to 58 TgCH(4) yr(-1), range 51-72, -14 %) and higher emissions in Africa (86 TgCH(4) yr(-1), range 73-108, + 19 %) than bottom-up values used as prior estimates. Overall, uncertainties for anthropogenic emissions appear smaller than those from natural sources, and the uncertainties on source categories appear larger for top-down inversions than for bottom-up inventories and models. The most important source of uncertainty on the methane budget is attributable to emissions from wetland and other inland waters. We show that the wetland extent could contribute 30-40% on the estimated range for wetland emissions. Other priorities for improving the methane budget include the following: (i) the development of process-based models for inland-water emissions, (ii) the intensification of methane observations at local scale (flux measurements) to constrain bottom-up land surface models, and at regional scale (surface networks and satellites) to constrain top-down inversions, (iii) improvements in the estimation of atmospheric loss by OH, and (iv) improvements of the transport models integrated in top-down inversions. The data presented here can be downloaded from the Carbon Dioxide Information Analysis Center (http://doi.org/10.3334/CDIAC/GLOBAL_METHANE_BUDGET_2016_V1.1) and the Global Carbon Project.


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10 Brilliant Event Ideas for College Students
Don't get scared if your event ideas suddenly become the 'biggest thing' on campus.
If you have a passion for event planning, I know ten brilliant event ideas that you can organize while in college. It is never too early to start getting experience as an event planner. What's more, you have an audience in college; your fellow students will be delighted to have a fun break from their studies.
The event ideas I have written in this article are easy to achieve if you are focused, so make a choice and start planning. There are new students who will be delighted to attend these events and connect with others.
Organize TEDx talks on Campus
There is always an audience for TED talks on campus, and the topic options you have are unlimited, so you can make it a regular campus event. Thankfully, the organizers of TED talks are always willing to support the program on campuses because that is where it started. So, visit the website and find out the information you need to get started.
Start a fiction writing competition
Give the upcoming writers a chance to test their skills at fiction writing. You can make it an open event for everyone who is interested in writing. Some college professors may also be interested in being the judges to determine winners at the event.
Organize local community service
Many college students may already be doing things such as cleaning up their environment, advocating greener methods of living, and other environmentally friendly practices. These students will be delighted to be a part of an organized group who regularly engage in awareness campaigns about keeping the environment clean and safe for people and animals. Your events can be about creating awareness regarding environmentally friendly practices, cleaning up the college environment, the beach and streets, setting a good example for the others to follow.
Start a Campaign to encourage the consumption of organic foods
There is so much publicity about eating healthy meals and the need to choose organic foods over others. Many college students are willing to try it but may have not had the opportunity on campus. You can organize a mini-market on campus where farmers near the college can bring their products to sell. This is an idea that should get much support from the college administration and huge patronage from students.
Organize creative contests for students
This will be a fun event. There are many students who have creative hobbies, such as making short videos with smartphones, creating themes, mobile wallpapers, or Memes online. You can create a contest for this group of students. For example, start a contest for short videos with the most creative theme; this can be done easily with a smartphone and good editing. This is another campus event idea that can become a regular thing and eventually your brand, so give it all you have.
Organize creative handcraft classes
Many of the things we discard such as old picture frames, fashion accessories, and home décor can be creatively redesigned and used again. Handcraft is a thriving business online; you can find amazing pieces for sale on websites such as Etsy and Instagram. This event will teach college students how to creatively restore or redesign different things that can be sold or used in their dorms.
Arrange cooking lessons for students
This event should be fun for all participants. This is what you can do to get started; find the common quick and affordable meals that can be quickly cooked on campus and organize an event to teach students how to cook these meals. You will be helping the participants save funds by cooking more meals instead of eating at restaurants in the college.
Organize a mentorship program for students
The freshman batch comes into the college with little idea of how to survive in the new environment. Don't allow them to learn from experiences which may not be too good. It is better if they have a mentor to guide them. Your mentorship program can pair new students with seniors who will show them how the college system operates.
Start college music festivals
Give local and upcoming bands a chance to perform before a crowd of college students. This is going to be a fun experience and a chance to unwind after long hours of studying. Turn it up a notch by inviting food and drink vendors for more entertainment while the music bands perform. This can become a regular event at your college.
Start a fitness club
It doesn't have to be about bodybuilding, you can focus on physical activities to help students stay active and control their weight. You can organize aerobics in the mornings, runs and hikes through trails on campus. Students will like this, it is a chance to exercise and connect with other students. Without such events, many students won't bother about exercising, so you will be doing a good thing in the college community.
Don't get scared if your event ideas suddenly become the 'biggest thing' on campus. There are smart ways to handle the crowd, you can use apps such as
to schedule events and sell tickets. The Meeti app provides a perfect way to ensure that everyone can buy tickets conveniently.
With Meeti, you can monitor the response of your audience regarding ticket sales with the live analytics feature on the app. Try it today, Meeti takes away the stress of ticketing and scheduling events, allowing you to focus on ensuring that your audience has a wonderful time at the event.
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Teaching A level Mathematics (TAM) is a sustained professional development course designed for teachers who are new to teaching A level Mathematics. It is for teachers in state-funded schools and colleges in England. All participating teachers must be teaching AS/A level or IB Mathematics throughout the academic year in which they complete the TAM course.
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Our March event takes place from 6.30pm on Tuesday, 8th March, and we're delighted to have as our special guest poet, novelist, musician and lecturer Anthony Joseph. Born in Trinidad he moved to the UK in 1989. His publications include Desafinado (1994), Teragaton (1997), The African Origins of UFOs (Salt, 2006) and Bird Head Son (Salt, 2009). His work has also been included in several anthologies, including Identity Parade (Bloodaxe, 2010), Red (Peepal Tree, 2010) and Black, Brown & Beige (University of Texas Press, 2009). He performs and lectures internationally and tours with his band The Spasm Band. Joseph lectures in creative writing at Birkbeck and is a AHRC scholar and doctoral candidate at Goldsmiths College.
Anthony will be joined by Rebecca Rouillard, Narayani Guibarra, Michelle Shine and Jenny de Montfort reading short stories, novel extracts and poetry.You'll find them all on stage at the RADA Foyer Bar in Malet Street, London WC1E 7JN. Admission is free but with a suggested donation of £3.50 to Oxfam. To secure your place, email us at email@example.com


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Helpful Hints:
Bring or wear a jogging or fleece outfit in the cooler months, it is always nice to put on something warm and cozy when you come out of the water.
You are also welcome to bring a lap blanket.
We provide warm coco and coffee upon your return it always seems to hit the spot and warms you right up.
We perform more manatee tours in the cooler months than any other time of the year. (manatee tours offered all year)
The water temperature never changes and is a constant 70-72 degrees. (spring fed waters maintain the temperature).
Our 3m wet suits will keep you warmer longer in the water. sizes toddler up to 5xL. Creating awesome buoyancy control for guest (effortless floating)
Age Restrictions: We do have age restrictions but for coast regulations which means a child 6 years and under must remain in a life jacket at all times, additionally we require 18 years and under to be with a parent or guardian, secondly 14 years and younger must remain with a parent in the water or on the boat at all times. From time to time we have babies that attend our tours parents usually switch out their time in the water or bring someone to stay with baby as a dry rider and watch the manatee interaction from the boat. Coast guard considers even the smallest of guest as one person please include every guest riding along on the boat when making your booking.
Weather Conditions: We never cancel a tour due to rain (lighting conditions might mean a delay )
Cool weather is great conditions for when your are swimming with manatees so we never cancel due to cool weather!
Snorkeling with the manatees
Copyright © 2020 Adventure Diving-SeaDaddys - All Rights Reserved.
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Fitness is more fun with friends, donchya think? It's also more fun when you know that you're making progress and positive changes.
I'm part of a 12-Week online fitness group and this week we're working on HIIT workouts. in case you don't know, HIIT means "High Intensity Interval Training" and basically is a quick workout with short, intense bursts of work with brief periods of rest in between. Sounds tough? That's because it is! But it's also super effective. In fact, unless my clients are training for a race, I would prefer they did HIIT exercise vs. long, endurance cardio sessions.
We're talking getting the same calorie burn as an hour of treadmill jogging -- but in 15 minutes. We're getting off the machines. Ending the slow and steady cardio. And really challenging our hearts. Wanna try?
P.S. If you can't jump or perform any of these, MODIFY!! Just do an easier version of the move but don't stand still. The secret is to moving with intensity - and your intense might look different than mine. And that's A-OK!
P.P.S. Wanna workout with me? My online group fitness coaching goes down monthly and I save 10 spots for new clients each month. To apply, please fill out this form.
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Wrath of the Righteous
Abyssal Pitborn with anger issues
Character Summary
- Unwilling to trust many people closely.
- Strong desire to gain revenge on demons / cultists due to torture
- Plagued by impulsiveness and often lashes at people out of anger issues
- Has accepted her abilities, but is tormented by the idea of not being in control
- Cynical about general human nature after being disowned
- Impulsive in helping the more innocent.
Originally born in Old Kenabres, Kaeilyn's childhood and early years were a mostly idyllic experience. While life was hard in general in Kenabres, being an child of an established family made her early years relatively painless, albeit somewhat suffocating. No matter where she went as a child, the ever watchful eye of an aunt, uncle, or elder cousin was always looming overhead.
Kaeilyn's birth was a tumultuous occasion. The faint smell of brimstone and her mismatched eyes hinted that not all was right with her blood. Her family looked on with a mixture of revulsion and sadness – the demonic attack that had injured her ( nth great grand ) father had spread its roots so far.
Her childhood and teenage years were relatively boring. Kaeilyn had few friends to speak of, save for her elder sister Aliera. Days passed endlessly, melting together in a sea of repetition, save for the occasional failed excursion to various libraries. Any attempt at leaving the watchful eye of her relatives resulted in swift, merciless punishment. Aliera would do what she could to help Kaeilyn, gathering various tomes and books to keep her mind occupied.
Kaeilyn and Aliera were almost as different as night and day. While Kaeilyn was frail and more interested in a book than another human being, Aliera loved to chatter and . They would spend long nights talking about their hopes and aspirations. There was something about Aliera's expression that always calmed Kaeilyn – maybe her radiant smile or her pure, hope-filled eyes. Most of all, Aliera lacked the small twinge of scorn on her face that her other relatives tried to hide.
For as long as she could remember, Kaeilyn held a deep fascination with the endless amounts of crusaders traveling through the city. She envied their freedom and courage, even more so after Aliera joined the ranks of the Iomedian paladins to fight the demonic scourge. They communicated by mail often, but she rarely got to see her beloved sister.
As a young adult, Kaeilyn found herself troubled by strange dreams. Visions of bloody, gaping maws, razor-sharp claws, and unearthly screams became nightly visitors. Her sleep and overall condition became troubling enough for her to seek guidance from higher powers. She found solace within Desna's embrace, the nightmares eventually becoming no more than a sour memory.
One fateful night after her 22nd birthday, Kaeilyn ventured outside of Kenabres to meet up with Aliera, who had sent her a letter, promising grand adventures. Outside the walls, the darkness seemed to swallow her up. She awoke as abruptly as she had fallen asleep, chained face down to a stone table. Hooded figures surrounded her, an eerie glow keeping their faces hidden in the early hours of twilight. The figures were silent, save one behind her, chanting in some unknown tongue. She felt the cold steel of a dagger press against her back and screamed. The tool carved every last detail of Sifkesh's unholy symbol into her, the chanting growing ever louder as the blood dripped down her back. A red glow filled the air, and suddenly – silence.
Kaielyn awoke to the dull, throbbing pain of her body. Sunlight streamed in through the cracks in the temple. Her body felt odd – heavy and foreign. Her hand brushed up against her forehead to find protrusions that extended and curled back as horns. She was speechless as she began to examine the rest of her body. Her skin had turned a dark black and was cool to the touch. She stood up, nearly falling over due to her newfound physique, but managed to balance herself with her new tail. She expected to feel the cool stone tile underneath her toes, but they had been replaced by hooves. A quiet laugh escaped her lips, quickly turning into full on hysterics as Kaielyn convinced herself that this was another nightmare. The laughter slowly faded and turned to sobbing as she saw the slaughtered remains of the cultists strewn about, their blood painting the ground a dull crimson. The pain of the wound on her back had shattered any delusions she may have had.
Leaving the temple, she saw the silhouette of a woman dissapear into the nearby forest. Broken and exhausted, she trudged her way back to the walls of Kenabres. Her family reacted in a manner that should have been all too obvious from the start. They feigned knowing her when she appeared on the doorstep to her home, demanding that she leave before the witch hunters were called. She abandoned her family name soon afterwards.
Currently, Kaeilyn has been recently employed by the owners of the Truestone Quarry as a caravan guard. The abandonment by her family and the torture by the cultists have caused her once shy personality to fester into a deep distrust of most others. She is quick to anger and slow to forgive, personality traits that have been reinforced by her abrupt change in lifestyles. Her recent excursions to the town of Zharech for gladiatorial combat as a means of relieving stress have only exacerbated the situation once she found that she could utilize her rage when fighting. Kaeilyn's physique now reflects her occupations as the muscle of many groups. Deep down, she still wants to be a crusader, fighting for good like her sister, but the harsh realities of Kenabres working class have all but destroyed those ideals.