text:
- Methodology
- Open Access
PCR-based methods for the detection of L1014 kdr mutation in Anopheles culicifacies sensu lato
© Singh et al; licensee BioMed Central Ltd. 2009
- Received: 09 March 2009
- Accepted: 14 July 2009
- Published: 14 July 2009
Anopheles culicifacies s.l., a major malaria vector in India, has developed widespread resistance to DDT and is becoming resistant to pyrethroids–the only insecticide class recommended for the impregnation of bed nets. Knock-down resistance due to a point mutation in the voltage gated sodium channel at L1014 residue (kdr) is a common mechanism of resistance to DDT and pyrethroids. The selection of this resistance may pose a serious threat to the success of the pyrethroid-impregnated bed net programme. This study reports the presence of kdr mutation (L1014F) in a field population of An. culicifacies s.l. and three new PCR-based methods for kdr genotyping.
The IIS4-IIS5 linker to IIS6 segments of the para type voltage gated sodium channel gene of DDT and pyrethroid resistant An. culicifacies s.l. population from the Surat district of India was sequenced. This revealed the presence of an A-to-T substitution at position 1014 leading to a leucine-phenylalanine mutation (L1014F) in a few individuals. Three molecular methods viz. Allele Specific PCR (AS-PCR), an Amplification Refractory Mutation System (ARMS) and Primer Introduced Restriction Analysis-PCR (PIRA-PCR) were developed and tested for kdr genotyping. The specificity of the three assays was validated following DNA sequencing of the samples genotyped.
The genotyping of this An. culicifacies s.l. population by the three PCR based assays provided consistent result and were in agreement with DNA sequencing result. A low frequency of the kdr allele mostly in heterozygous condition was observed in the resistant population. Frequencies of the different genotypes were in Hardy-Weinberg equilibrium.
The Leu-Phe mutation, which generates the kdr phenotype in many insects, was detected in a pyrethroid and DDT resistant An. culicifacies s.l. population. Three PCR-based methods were developed for kdr genotyping. All the three assays were specific. The ARMS method was refractory to non-specific amplification in non-stringent amplification conditions. The PIRA-PCR assay is able to detect both the codons for the phenylalanine mutation at kdr locus, i.e., TTT and TTC, in a single assay, although the latter codon was not found in the population genotyped.
- Indoor Residual Spray
- Voltage Gated Sodium Channel
- Amplification Refractory Mutation System
- Surat District
- Anopheles Culicifacies
Anopheles culicifacies s.l. is the main malaria vector in the Indian subcontinent, affecting mainly rural areas, and contributes 60–65% of malaria cases in India [1]. As this is an endophilic vector, indoor residual spraying (IRS) of DDT is the main strategy used for its control. This species is resistant to DDT in most parts of India. Although DDT is banned in many countries, the recent endorsement by World Health Organization for the use of DDT for IRS for malaria vector control [2] has renewed interest in this insecticide. Pyrethroids are the most commonly used insecticides for IRS and the only insecticide class recommended for impregnation of bed nets due to their relatively low mammalian toxicity and rapid knock down effect on insects. In India, the use of pyrethroids was initiated in 1990s to control malaria epidemics in areas where An. culicifacies s.l was resistant to DDT and malathion. Pyrethroid resistance in An. culicifacies s.l. was detected in Surat district of Gujarat state, western India, soon after its introduction in vector control programme [3].
DDT and pyrethroids are neurotoxins that act on the voltage-gated sodium channels by modifying their gating kinetics, resulting in the prolonged opening of individual channels leading to paralysis and death of the insect. One of the mechanisms of pyrethroid resistance in insects is referred to as knock-down resistance (kdr) caused by reduced target site sensitivity. The phenotype is commonly conferred by a single point mutation (L1014F/S/H) in the IIS6 segment of voltage gated sodium channel [4, 5]. Other mutations in different regions of the gene also confer knock-down resistance in some insects [4, 6], but among anophelines this is the only locus where point mutations have been reported to date conferring resistance. Only two points mutations have been reported in anophelines at this locus–L1014F in Anopheles gambiae [7] (West African kdr), Anopheles arabiensis [8] and Anopheles stephensi [9], and L1014S in An. gambiae [10] (East African kdr).
The Leu-Phe mutation at the kdr locus in An. culicifacies s.l. was reported by Hoti et al [11] using an allele-specific PCR assay, whose external primers Agd1 and Agd2 were based on An. gambiae sequences. However, authors failed to amplify the DNA region of interest using the primers Agd1 and Agd2 (see Additional file 1) due to mismatching with template DNA. As the evident cross resistance between DDT and pyrethroids in this resistant strain strongly suggested a kdr type phenotype, an attempt was made to confirm the presence of kdr-based insecticide resistance in An. culicifacies s.l. by DNA sequencing and developing alternative high throughput methods for kdr genotyping. The part of the voltage gated sodium channel spanning IIS4-IIS5 linker to IIS6 segments of An. culicifacies species A, B and C was first sequenced which contains at least five residues where mutations have been reported in other insects, namely, Met918 in the IIS4-IIS5 linker, Leu925, Thr929 and Leu932 in IIS5 and Leu1014 in IIS6 [6]. Based on the sequences obtained, three PCR based assays for kdr genotyping were developed, i.e., Allele Specific-PCR (AS-PCR), Amplification Refractory Mutation System (ARMS) and Primer Introduced Restriction Analysis-PCR (PIRA-PCR). These assays were tested on a DDT/pyrethroid resistant An. culicifacies population from Surat district and validated through DNA sequencing. The ARMS and PIRA-PCR were developed as alternatives of AS-PCR, which is often alleged to be non-specific [12–14]. Additionally, the PIRA-PCR was developed as a method by which both the codons of Phe (TTT and TTC) at kdr locus can be detected by a single assay.
Study area
Anopheles culicifacies s.l were collected from the villages of Surat district of Gujarat state (21–22°N latitude and 73–74°E longitude) located within three Primary Health Centres (PHC), Amladam, Bareda and Dadheda, where a significant level of resistance to DDT and pyrethroids (7–13% and 60–69% mortalities recorded in adults against 4% DDT and 0.05% Deltamethrin, respectively, in WHO's standard insecticide susceptibility tests) have been reported [3]. This An. culicifacies s.l. population is comprised of species B and C [3]. Indoor resting mosquitoes were collected using hand aspirators between 0600 and 0800 AM. The mosquitoes were preserved in isopropanol and DNA from individual mosquitoes was isolated.
DNA sequencing
cDNA sequencing
The total RNA of An. culicifacies species A, B and C was isolated using TRIzol Reagent (Invitrogen Corporation) according to the manufacturer's instructions. cDNA synthesis and RT-PCR were carried out using AccessQuick RT-PCR System (Promega Corporation). The first strand cDNA was synthesized by avian myoblastosis virus reverse transcriptase (AMV-RT) at 45°C for 45 min, followed by denaturation of RNA/DNA hybrid and inactivation of AMV-RT at 95°C for 2 min. Second strand cDNA synthesis and subsequent DNA amplification were done by Tfl DNA polymerase at the following conditions: 35 cycles of denaturation at 95°C for 30 sec, annealing at 45°C for 30 sec, and extension at 72°C for 1 min followed by a final extension of 5 min at 72°C. The primers used for RT-PCR were Kdr1F and Kdr1R. The product was re-amplified by semi-nested PCR using primers Kdr1F and KdrR, and thereafter sequencing was performed using BigDye Terminator v3.1.
The DNA sequences obtained from cDNA and genomic DNA analyses were deposited in GenBank (accession numbers: GQ268331–GQ268333 and GQ279245–GQ279247).
PCR based assays
Three molecular methods for kdr genotyping were developed based on sequences obtained in this study, viz., Allele Specific PCR (AS-PCR), Amplification Refractory Mutation System (ARMS) and Primer Introduced Restriction Analysis PCR (PIRA-PCR) (Figure 1). In all the PCR-based assays, unless otherwise stated, uniform concentrations of reagents, except primers, were taken, which were: 1× Buffer, 1.5 mM of MgCl2, 200 μM of each dNTP and 0.5 unit AmpliTaq Gold® per 15 μl of reaction mixture. All the PCR reactions were carried out in a GeneAmp® PCR System 9700 (Applied Biosystem) thermal cycler. The three assays were tested on field collected An. culicifacies s.l for kdr genotyping and results were validated following DNA sequencing.
Allele-specific-PCR (AS-PCR) analysis
Amplification refractory mutation system (ARMS)
To check the specificity of ARMS, the assay was also performed with increased concentration of MgCl2 (up to 3.0 mM) and AmpliTaq® Taq DNA Polymerase (up to 2.5 units per 15 μl reaction mixtures) at an annealing temperature of 45°C.
Primer-introduced restriction analysis-PCR (PIRA-PCR)
PIRA-PCR is based on incorporation of a deliberate mismatch in one primer so that a new restriction enzyme recognition site is created in the PCR amplicon in the presence of a specific allele in the target DNA. A PIRA primer KdrP (forward) with sequence 5'-TCC TGG CTA CAG TAG TGA TAG GAA AaT T-3' was designed, where a T-to-A mismatch was incorporated at the 3rd base position from 3' end terminus (shown in small letter in primer sequence). The terminal AAaTT-3' sequence of the PIRA primer (KdrP) creates a recognition site for the restriction enzyme Apo I or Xap I, i.e., AAATTY, in the amplified PCR product with the primer KdrR (reverse) in the presence of the Phe codon (TTT or TTC) in the template DNA at the kdr locus. Hence the PIRA-PCR is capable of detecting both possible codons of phenylalanine (TTT or TTC). With the Leu (TTA) allele, no Apo I restriction site is created.
The predicted size of PCR product amplified with primers KdrP and KdrR is 134 base pairs, which when digested with ApoI or XapI, will be cleaved into 106 and 24 bp fragments (with four bases 5'-overhang) in the presence of Phe allele (TTT/TTC). No cleavage is expected to take place in presence of Leu (TTA) allele. The 134 and 106 bp fragments can be easily resolved on 3% agarose gel whereas 24 bp fragment is normally not expected to be visualized or difficult to discriminate from primer-dimer. Therefore, 134 bp band was scored as S and 106 bp as R.
Validation of PCR based assays
All the mosquito samples genotyped as RS (n = 18) and RR (n = 1) were sequenced alongside 23 SS in order to validate assays and to check for other mutations in addition to the Leu-Phe change. The primers used for PCR amplification and sequencing were KdrF and KdrR. The sequence data were aligned and analysed using Mega 4 alignment utilities [18].
Hardy-Weinberg equilibrium analysis
The Exact Test of Hardy-Weinberg equilibrium using a Markov chain on three genotypes, i.e., SS, RS and RR was done using software Arlequin ver 3.11 [19]
DNA sequencing revealed the presence of a Leu (TTA) to Phe (TTT) mutation at residue L1014 in a total of 19 samples, 18 of which were heterozygote and one homozygous resistant individual. No other mutations were recorded, including L995H mutation reported in Iranian An. culicifacies s.l. [20].
Testing of AS-PCR and ARMS assays on An. culicifacies s.l. samples from Surat revealed presence of discrete bands of expected sizes, as visualized on agarose gel. With PIRA-PCR also all the expected bands were seen, except 24 bp fragment resulting from the cleavage of PCR product (134 bp). Therefore, the presence of 106 bp cleaved product was scored as R. A total of 186 samples were genotyped using AS-PCR, of which 167 were assigned as homozygous susceptible (SS), one as homozygous resistant (RR) and 18 as heterozygote (SR) based on the criteria mentioned in section 'Methods'. All of the RS and RR, and 80 of the SS samples, as genotyped by AS-PCR, were also assayed with ARMS and PIRA-PCR assays, with similar results to the AS-PCR.
The result of DNA sequencing of 42 samples genotyped by all the three assays (RR = 1, SR = 18, SS = 23) were in agreement with PCR based assays. Thus all three assays are specific and can be used for genotyping.
The AS-PCR, based on a single mismatch at the 3' end of the allele specific primer, was specific and no mispriming was observed. The ARMS was specific even with increased concentrations of MgCl2 (3.0 mM) and taq polymerase (3 Units of AmpliTaq Gold per 25 μl reaction).
The observed and expected heterozygote frequencies were 0.09677 and 0.10202 respectively, which did not differ significantly (p = 0.41220) from Hardy-Weinberg equilibrium.
Malaria control in India relies on control of the major vector An. culicifacies. Knock-down resistance in An. culicifacies may be a serious threat to the continued use of pyrethroids, which are currently the insecticide class of choice for control of multiple insecticide resistant vector population by IRS and the only insecticide class available for the impregnation of bed nets. The present study provides the first direct evidence of the presence of the L1014F kdr mutation in Indian An. culicifacies.
The point mutation at residue L1014 leading to Leu-to-Phe change in voltage gated sodium channel is a most common mechanism associated with knock-down resistance in insects against DDT and pyrethroids. Variant replacements of this residue, L1014H and L1014S, have also been reported [4]. In anophelines, only two mutations, L1014F and L1014S, are reported conferring resistance [7–10]. No mutation at other location has been reported in anophelines except a novel mutation, L995H, reported in a single specimen of An. culicifacies s.l. from Iran [20]. During this study, screening of limited numbers of An. culicifacies s.l. samples from Surat didn't revealed presence of L995H mutation. The implication of such mutation in DDT/pyrethroid resistance and frequency in resistant population is yet to be investigated.
For monitoring of kdr resistance in vector population, there is a need of simple and reliable methods. AS-PCR is a convenient and inexpensive method used for kdr detection. This assay format has widely been used with the malaria vector An. gambiae in Africa [7, 10]. For An. culicifacies s.l., an AS-PCR was designed by Hoti et al [11] using the external primers Agd1 and Agd2 originally designed for An. gambiae. These primers do not work with An. culicifacies s.l. from Surat district (see Additional file 1) due to mismatch with template DNA which was revealed after sequencing presented in this study and, therefore, not reliable. Further, a significant departure from HW equilibrium, resulting from deficiency of heterozygotes, had been noticed by the authors, while re-examining genotyping data of kdr in An. culicifacies s.l. by Hoti et al [11], which may be due to discrepancy in genotyping result. This necessitated the development of culicifacies-specific molecular assays for kdr genotyping.
The AS-PCR designed for SNP detection is based on single base primer-template mismatch at 3' of the primers. These primers are generally specific but in some instances the mismatched base does not prevent the extension process [12, 13]. Mispriming with the primer specific for susceptible (kds) allele has also been reported in An. gambiae [21], where the mismatched base is at penultimate position from the 3' terminus. To avoid such mispriming, an ARMS method was developed, where an additional deliberate single nucleotide mismatch is incorporated near the 3' end of the allele-specific primers, which increases the specificity of the ARMS primers [16].
PIRA-PCR is a very reliable method for SNP detection that reduces the possibility of genotyping error due to mispriming. The PIRA-PCR was initially used for genotyping of kdr in An. gambiae by Janeira et al [22], where the mismatch in the PIRA primer was incorporated at the penultimate 3' base in order to create a restriction site. In the PIRA primer designed here for An. culicifacies s.l., the mismatch was incorporated at the third base position from 3' end, which seems to be advantageous, being less refractory to amplification in comparison to those where mismatch is at penultimate base. One of the most significant advantages of current PIRA-PCR over previously described methods is its capability to detect both possible codons of phenylalanine, i.e., TTT and TTC, whereas other PCR based assays can detect only one codon. Although in the single mosquito population tested only the A-to-T substitution was encountered, which is the most common substitution in mosquitoes [5], the possibility an A-to-C substitution in other mosquitoes cannot be ruled out and has recently been seen in Culex quinquefasciatus where both substitutions (A-to-T/C) exist [23]. It is possible that the A-to-C substitution is being overlooked in other species. The PIRA-PCR strategy described here could be used for screening of Leu-Phe based kdr for other mosquito species with little modification in primer sequences. The PIRA-PCR is relatively costly compared to AS-PCR or ARMS, due to involvement of a restriction enzyme step, and it is also slightly more time consuming. The PIRA-PCR may be misleading in case of incomplete digestion of amplified product where RR samples may be scored as RS.
Recently several other molecular methods in addition to PIRA-PCR have been used for kdr genotyping as alternative of AS-PCR such as Heated Oligonucleotide Ligation Assays (HOLA) [24], Sequence-Specific Oligonucleotide Probes (SSOP) [25], Fluorescence Resonance Energy Transfer/Melt Curve Analysis (FRET/MCA) [26] and fluorescent based genotyping by capillary electrophoresis [27], but most of these methods are expensive or require more sophisticated instruments. PCR-based methods on the other hand are easy, inexpensive and can be performed in the most of the laboratories equipped with Thermal Cycler.
The Leu-Phe mutation (L1014F) in the voltage gated sodium channel, which generates the kdr phenotype in many insects, was detected in a pyrethroid and DDT resistant An. culicifacies s.l. population from Surat, western India. Three PCR-based methods (AS-PCR, ARMS and PIRA-PCR) were developed for kdr genotyping based on DNA sequences obtained from An. culicifacies species A, B and C. All the three assays were specific. The ARMS method was refractory to non-specific amplification in non-stringent amplification conditions. The PIRA-PCR assay is able to detect both the codons for the phenylalanine mutation at kdr locus, i.e., TTT and TTC, in a single assay, although the latter codon was not found in the population genotyped.
This collaboration was funded by the Wellcome Trust. Authors are thankful to Mr Uday Kumar and Mr Shri Bhagwan for excellent technical assistance.
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The US government now reportedly believes that North Korea is directly to blame for the devastating and unprecedented attack on Sony, The New York Times reports.
The massive Sony data hack is likely retaliation for the controversial comedy "The Interview," which depicts the assassination of North Korean dictator Kim Jong Un.
US investigators have now parsed through enough evidence to claim that North Korea is "centrally involved."
An unnamed government official told NBC News that the US has found "linkage to the North Korean government" although they believe the hack originated outside of the country. The US reportedly believes that the hackers were acting on orders from North Korea, although it's unclear what evidence investigators have that points to that.
There is reportedly a debate within the White House about how to respond now that the government believes North Korea is to blame for the hack, which exposed a massive amount of emails, employee information including salaries and medical history, and business documents from within Sony.
A hacking group calling themselves the "Guardians of Peace" has claimed responsibility for the attack and posted an online threat Tuesday against theaters planning to show "The Interview." The threat promised a "bitter fate" for those who went to see the movie.
US security authorities said Tuesday that they were investigating the threat but had so far seen no real sign of an active plot. A second US official said Wednesday that investigators are not aware of any people or entities capable of actually carrying out such threats against cinemas or audiences.
But theaters are still taking precautions. "The Interview" was scheduled for general release on Dec. 25, but Sony canceled the release of the movie late Wednesday afternoon after many large cinema chains said they wouldn't show the film.
In light of the cancelation, Hollywood has not been quiet. Many have argued that the studio gave into the hackers and violated the First Amendment.
North Korea complained to the United Nations about "The Interview" in June, accusing the US of sponsoring terrorism and committing an act of war by allowing the movie's production.

subset: commoncrawl


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Eurozone: A very overcast outlook
The eurozone is back in recession.
Economic activity declined in the third quarter of the year, according to new data from the EU's statistical agency, after a contraction in the previous period too.
In fact the eurozone has been very close to recession for a longer period.
In the first quarter of this year, there was no growth and a decline at the end of 2011. So it's a year since the eurozone's economy grew at all, and you have to go back to the beginning of last year to find a figure that would, under more normal economic circumstances, have been considered reasonable growth (0.6 %).
Another indicator of the depth of the Europe's problems is the fact that the recovery from the last recession lasted less than three years.
This weakness is, up to a point, consistent with the pattern that influential academics have found in the period after financial crises. Carmen Reinhart and Kenneth Rogoff say that the recovery from a recession caused by a financial crisis tends to be relatively slow.
Weakness in Europe
In the case of Europe, the financial crisis - that's to say, the extent to which the problems were rooted in the banking system - was particularly acute in the case of Spain and Ireland. Booms in lending and property prices were followed by sharp retrenchments and bank failures.
But across the eurozone banks were hit by investments in flawed financial assets or by lending to financially-stressed governments.
What this means is that some degree of weakness was to be expected in Europe.
But has it been aggravated by austerity, the spending and taxation policies pursued by many European governments seeking to reduce their borrowing needs?
Professor Paul de Grauwe, of the London School of Economics, says the eurozone is in a "double-dip recession which is entirely self-made". He blames what he calls "excessive austerity in southern countries".
The idea that the austerity has deepened Europe's downturn is widely shared. It is another question whether there is a realistic alternative.
Several eurozone governments have debt burdens that are rising unsustainably. There is a debate to be had, however, over how rapidly they should do that. The International Monetary Fund has suggested there is a case for moderating the pace of cuts.
The countries in the most acute financial and economic difficulty also have problems with competitiveness. Membership of the euro means they don't have the option of allowing their currency to depreciate to readdress that problem. Instead, they have been hoping that reforming their economies, notably their labour markets, would do the job.
There are signs of progress in that area.
In the case of Greece, the EU's commissioner for economic affairs, Olli Rehn, recently said "by the end of this year, all of the competitiveness loss experienced by Greece between 2001 and 2009, relative to the rest of the euro area, will have been recouped".
It is nonetheless a process that takes time, much more time than a currency depreciation.
Worse ahead?
Looking ahead, many economists expect worse to come. Surveys of business have suggested a further deterioration is likely, even in Germany. Growth there has already slowed. The most recent figure was a decidedly sluggish 0.2% in three months. If Germany were to slip into recession, that would be a powerful symbol of the eurozone's problems.
It would also be another headwind to recovery. As well as being an exporting powerhouse, Germany is also a major buyer of goods and services from the rest of the eurozone.
It is, then, a very overcast outlook. To take one example, the London-based consultants Capital Economics forecasts a contraction of 2.5% for the eurozone next year.
The European Commission is less pessimistic. Its recent forecast has the eurozone growing in 2013 year. But the figure is a frankly pitiful 0.1%, followed by a still rather lacklustre 1.4% the following year.
Even if the Commission turns out to be right, that would be a very long period of sub-par performance.


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This is a Service Pack release of the MySQL Enterprise Server 5.0.
This section documents all changes and bugfixes that have been applied since the last MySQL Enterprise Server release (5.0.74).
If you would like to receive more fine-grained and personalized update alerts about fixes that are relevant to the version and features you use, please consider subscribing to MySQL Enterprise (a commercial MySQL offering). For more details please see http://www.mysql.com/products/enterprise/advisors.html.
Functionality Added or Changed
Bugs Fixed
Functionality Added or Changed
libedit library was upgraded to version 2.11. (Bug #42433)
Bugs Fixed
An attempt by a user who did not have the
SUPER privilege to kill a system thread could cause a server crash. (Bug #43748)
Tables could enter open table cache for a thread without being properly cleaned up, leading to a server crash. (Bug #42419)
The SSL certificates included with MySQL distributions were regenerated because the previous ones had expired. (Bug #42366)
Some queries using
NAME_CONST(.. COLLATE ...) led to a server crash due to a failed type cast. (Bug #42014)
DATE_FORMAT() could cause a server crash for year-zero dates. (Bug #41470)
SET PASSWORD caused a server crash if the account name was given as
CURRENT_USER(). (Bug #41456)
When substituting system constant functions with a constant result, the server was not expecting
NULL function return values and could crash. (Bug #41437)
Creating a table with a comment of 62 characters or longer caused a server crash. (Bug #39591)
EXPLAIN EXTENDED evaluation of aggregate functions that required a temporary table caused a server crash. (Bug #34773)


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Dear August
On behalf of the Council of the Dramatists Guild
We read with great sadness of your illness. As your fellow writers, we wanted to make sure you knew how valuable you are to us, and how immeasurable your contribution has been both to the American theatre and the American culture as a whole. In pledging yourself to complete such a profound body of work, you have challenged us all to get our work done, without succumbing to the distractions of the commercial world, or the voices of the critics. Your work reminds us to tell the stories that only we know, to face the fury in our lives, and seek justice and understanding for our characters, saints and sinners alike. You have shown what can be accomplished by demanding that the alien world make a place for you.
Your plays have revealed an America that many of us would not otherwise have known. You have allowed white Americans to look behind the images we see of black culture, and you have given black Americans a chance to see their families and forbears onstage. You have written a history that would have remained unknown were it not for you, you have opened doors that still threaten to keep us living in separate rooms in this country. You have paved a road for travel by young black writers, and you have changed forever the way black Americans are portrayed onstage and understood in the world.
We collected a group of letters to send to you, then decided they might be too daunting for you to read at this point. But here are some excerpts, offered without attribution, and accompanied by our deepest prayers for your peace and comfort.
"August Wilson is the epitome of the men who people his plays. Strong, passionate, honorable, stubborn and good. I have recognized my father, my mother, my sister and brother in his characters. I'm grateful to him for showing me his poetic vision of a world in which I had always belonged but had never appreciated for its true value."
"August Wilson has crawled into the soul of a woman, and written the most thrilling arias for women I have ever heard."
"August Wilson's stories have enriched my life and my understanding of the world. His people have entered my consciousness. I hear them talk, watch them move, in my mind's eye now as I write this, like every vivid personality I've encountered in "real" life.
"August Wilson's contribution to the American Theatre has been profound, both as a playwright and a builder of audiences."
"His characters have lives and histories that go back for decades, generations — and the depth and complexity of their entanglements and connections is apparent in every gesture, as rich as life."
"My students are stunned to encounter the verbal pyrotechnics, and the surprises. Oh the surprises."
"…in the audience for an August Wilson play, people sit up and pay attention. They know they are in for something special. As the play unfolds, you can feel the temperature as he introduces a threatening character or peculiar behavior."
"August Wilson's characters appear at the moment at the moment in their lives when everyone is changed forever. He is a beacon of truthful, powerful storytelling."
"August Wilson has been building a bridge, one span at a time. In the end, he will be able to open the entire structure and we'll be able to cross, experiencing a hundred-year journey that will provide understanding not only for African-Americans but for all who are grappling with our lives in this troubled country."
"His work is a treasure, his writing is poetic, mystical and startling I want to thank August Wilson for the most heart wrenching moments I've had in the theatre."
"I loved his guts and candor. He wasn't scared of anybody. I wish I had some of that."
These notes go on and on, but the thoughts are the same – an outpouring of gratitude and yes, love, from people who have watched you and learned from you and counted on you for more laughing, more singing, more sex, more truth – more August Wilson.
I remember the day you acted as my escort, walking me up to the podium for a Pulitzer Prize celebration that New Dramatists was putting on. That was before you'd won both of yours, of course. But I remember the cool, noble guy who for that moment, made me feel able to give the keynote. I thank you for that bit of comfort, and on behalf of my students, I thank you the world you have preserved, the lives you have created, the vision you have shared, the example you have set. I wish we had had more conversations, but in their place, I have all these plays of yours, to read and read for the rest of my life. If I can do anything else, please let me know. I am your grateful friend,
Marsha Norman


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In August 1987, several Darwin Institute of
Technology staff from the DIT Students Association and the then Director of
Coaching for the Northern Territory Rugby Union discussed the possibility of
DIT entering a C-Grade team into the 1987/88 Rugby season. Following key
signings of a few local stars, the Pirates were then convinced to field an
A-Grade side as well. The DIT Pirates Rugby Union Football Club was formed and
the Pirates were set to become a reputable force in Darwin Rugby Union.
From the outset, the members of that fledgling Club strove on the playing fields to be competitive, attempting to align themselves with the on-field tactics displayed by the top Rugby Union sides of their era. It was not merely that they were able to imitate their strategies, but more that they set themselves ambitious game plans and tried hard to achieve them. The style of game they set for the Club quickly became its trade mark.
Today, the University Pirates Rugby Union Football Club is based at Charles Darwin University in the Northern suburbs of Darwin in the Northern Territory. University Pirates is now a well established club, consisting of 75 senior players and 125 juniors. The club also boasts a modern club house situated on a picturesque oval with newly built changing rooms and state of the art gym facilities. The club is very much family orientated with various activities for the juniors during home games and training sessions including family dinners on Tuesday and Thursday nights.
As was intended back in 1987, the Pirates of today continue to play with ambition, determination, and a winning team attitude. As a testament to this, the University Pirates have gone on to win 9 premierships since their inauguration into the A grade competition and have been well represented in 16 hard fought grand finals. Pirates players wear the uniform with pride and play with the badge aptly above their hearts; giving their all for their team mates, and driving forward in pursuit of victory.
University Pirates RUFC - "Never to be crossed"


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The decisions should be delivered to the banks (branches of banks) in electronic form through the Interregional Inspection of the Federal Tax Service for the central data processing unit, authorized by the Bank of Russia and and local Central Bank branches.
Electronic message should include: the decision of the Tax Authority in electronic form, encrypted package, confirmation, notice, receipt notification, which constitute the complete package of information, compiled under stadard format, suitable for clear perception, supplied with authentication ID (hereinafter ID) for control the integrity and authenticity of information.
The procedure of the encrypted electronic messages delivery from FTS through the internal banking system enables prompt counteraction between tax authority, RF Federal Tax Service and the bank (bank branch) directly, without involvement of the Bank of Russia.
3. The RF Ministry of Finance letter N 03-05-04-01/32, dated 13.10.2008 and RF MF and FNS letter N SHS-6-3/787 @, dated Oct. 31, 2008 clarified the tax rates, applicable to the property of individuals, applying special tax regimes.
In particular, it is noted, that with regard to Art. 18 of the Tax Code, special tax regimes, stipulated therein, provide for exemption from certain taxes and levies. But the benefits, provided to a taxpayer in regard to property tax on individuals due to special tax regimes, do not result in exclusion those assets from the relevant tax base. That is, the property tax on business, operating under special tax regime, is subject for assessment, rather than payment in this case.
4. The RF FNS letter N SHS-6-3/765 @, dated October 24, 2008, has clarified the issue of corporate income tax assessment. A taxpayer has the right to transfer to tax assessment on the basis of monthly advance payments, rather than actually amounts received, by notifying the tax authority no later than December 31 of the year, preceding to the tax reporting period, when the transfer under advance payment regime is made. Herewith, regime of advanced payments should not be changed by the taxpayer within the complete tax reporting period.
5. The RF Ministry of Finance letter N 03-02-07/2-192, dated of October 29, 2008 has clarified the issue of penalties, accrued by tax authorities after the term of arrears recovery has expired.
In accordance with Article 46 of the Tax Code, the term (the so-called preclusive time frames) is set up, within which the tax authority should make the decision on tax arrears recovery instructions to the bank to transfer the tax due to the RF budget. Enforcement of penalties payment should not be considered in isolation from the duty to pay taxes, according to the RF Ministry of Finance. In the event the tax authorities miss the deadline for sending the recovery demand to the bank, all arrears and penalties should be claimed through the court. Herewith, if the preclusive term for the tax recovery is overdue, penalties can not serve as the means to ensure tax payment, and from that point, there are no grounds for charging thereof. The same approach is expressed in the Resolution of the Supreme Arbitration Court Board of the Russian Federation N 8241/07, dated of 06.11.2007.
6. In view of the adoption of the Federal Law N 175-FZ, dated of October 27, 2008 On additional measures to strengthen the stability of the banking system up to 31.12.2011, one can expect a large number of disputes and lawsuits.
Thus, it is foreseen, that the Deposit Insurance Agency should provide assistance to the investors and banks. The presence of an intermediary service in the person of the Agency in the transfer of funds from the federal budget to the banks and investors in the form of assistance will entail excessive tax losses and result in reduction of the investors' interest in the acquisition of insolvent banks.
From the standpoint of the Tax Code, assistance in the form of monetary transfers is a free-of-charge income of investors, i.e., it is subject to profit tax in the total amount. However, the Agency has no right to transfer funds for the investorТs tax, i.e., the investor would have to spend all assets, received from the Agency for the purpose of preventing bankruptcy of insolvent bank, and moreover, to pay income tax at the expense of its own funds.
There is also a difficulty in the fact, that the Agency has granted the right to invest the assets of physical entities in deposit insurance fund to finance activities on prevention the banksТ bankruptcy (Clause 3, Art.10). As far as the Fund is supposed to perform targeted activities, payments, addressed to other purposes than deposit insurance, can be recognized as the misuse of assets (though legitimate in terms of the federal law).
Such payments of the Agency can be subject to profit tax, as in accordance with the Tax Code (Article 321, Para.3), only obligatory and non-refundable expenditures are relevant to reduce the tax base of the organization only by binding costs. Since the action plan on bankruptcy prevention should be developed individually for each bank and approved by the Board of Directors of the Agency, those expenses cannot be considered as binding ones (that is, Para.3, Art.321 is not applicable), and. as they are not related to payments for the goods (works, services), they can not be applied to the Agency tax base reduction as other ordinary business expenses.
Apparently, problems will arise in regard to compulsory reduction of the authorized capital of problematic bank. In the framework of the federal law N 175-FZ, the RF CB has already adopted Regulation N 2108-U of 29 October, 2008 On the Order of adoption the RF CB resolution to reduce the bank's authorised capital, equal to the value of its own funds (capital). In fact, in the balance of the bank founders contributtions to the authorized capital of the bank are reflected in the amount of the original actual cost. However, the banksТ founders are not necessarily credit organizations or legal entities. I think that compulsory reassessment on the grounds of the RF CB resolution the balance sheet assets of on-banking organizations and contributions of legal and physical entities to the authorized capital of commercial banks, reflected in the Articles of Incorporation, may also provoke legal disputes.
The above analysis of the tentative disputable issues is preliminary and perhaps other types of lawsuits and disputes can arise in future.
7. The RF Federal Tax Service Letter N SHS-6-3/782 @, dated October 29, 2008, provides detailed comments on payment procedure for the VAT tax agents, transferring the funds for the works (services), supplied in the territory of the Russian Federation, purchased from the taxpayers - foreign entities, who are not registered with the tax authorities as taxpayers (Par.4 of Article 174 of the Tax Code).
8. The RF FTS Letter 2008 N SHS-15-7/1275 as of November 1, has clarified the FTS approach to social deductions, agreed with the Authorized Representative of the Russian Government to the Constitutional Court of Russia, the Supreme Court and the Supreme Arbitration Court. It is clarified, that Art.219 and 210 of the Tax Code provide the right to taxpayers to receive social tax deduction in the amount paid by the taxpayer in the tax period for his/her study courses in any in educational institutions, regardless of the type of training of the taxpayers, as well as the right to taxpayers-parents to receive social tax deduction in the amount paid by them for the education of their children in the form of full-time attendance.
The RF FTS Russia does not consider the limitation in regard to the form of the children full-time attendance as contradictory to the Russian Constitution and commented, that the author of federal legislation has the right, based on the principle of fairness and taking into account the economic, social and other conditions, set different tax requirements to various categories of taxpayers (RF Constitutional Court Resolution N 14-P, dated 16.07.2004). In fact, tax exemption is a benefit in nature, which is an exception from the Russian Constitution principles of universality and equality of taxation (Arti cles 19 and 57). Therefore, benefits should always be targeted, and establishment thereof is an exclusive prerogative of the legislature.
Social tax deduction in the amount paid by the taxpayer-parent for the education of their children is provided for the amounts, paid by the taxpayer to the third parties (not to themselves) without restrictions in the children age (up to 24 years). Those who are studying in the full-time form, have no opportunity to work and, respectively, to pay for their education (which implies the need for the parentsТ assistance). Therefore, imposing the limiting requirement in the full-time form of education for social tax deduction does not violate the principle of universality and equality of taxation, because it takes into account a legally relevant and objective distinction between individual categories of taxpayers.
9. The Letter of the RF Ministry if Finance and Federal tax Service N SHS-6-3/793 @ of 1 Nov., 2008 provides clarifications on the specifics in treatment for the tax purposes the banksТ profit, derived from interest on debt liabilities in cases, where the contract provides for the possibility of change (replenishment or withdrawal) of the bank deposit within the contract term.
In accordance with Article 269 of the Tax Code, in regard to such the debt liabilities, (in case there are no provisions for the interest rates amendments throughout their validity term), the RF CB refinancing rate, effective on the date of monetary assets involvement is established. Therefore, if the agreement provides only for the changes in the amount of the deposit, rather than in interest rates, in order to assess expenses for taxation purposes, the refinancing rate, set up by the RF CB on the date of the contract conclusion should be applied.
10. The RF Government Resolution N 833, dated of November 7, 2008 introduced amendments to the Regulations for approval the scope of minerals losses in the mining industry, based on the procedure and technology of the deposit development, approved by the RF Government Resolution N 921, dated of December 29, 2001. Standard losses rates for solid minerals are assessed individually in view of their location, technology of mining operations and should be included in the project documentation (technical design, feasibility study, deposit development project and other documentation).
Loss standards of solid minerals (rare ones) are approved by the Federal Agency for Subsoil Use.
The loss standards for common minerals are approved by public authorities of the subjects of the Russian Federation. Standards in losses of hydrocarbons are approved by the RF Ministry of Energy (annually).
Upon the approval, the responsible authorities provide the details on the upgraded standards to the RF RTS within 10 days.
11. By the Order of the Ministry of Economic Development N 392 of Nov. 12, 2008, the deflator K1 at the rate of 1.148 is established for 2009, which is applied to assess the tax base for the unified tax on imputed income in accordance with Chapter 26.3 of the Tax Code "Taxation system in the form of a unified tax on the imputed income for certain types of activities".
12. By the Order of the Ministry of Economic Development N 395 of Nov. 12, 2008 the deflator index at the rate of 1.538 is established for 2009 in order for application Article 26.2 of the Tax Code "A simplified taxation systemФ.
13. Federal Law № 205-FZ of 24.11.2008 has made amendments to the RF Budget Code and the RF Tax Code. In particular, the structure of income taxes to the budgets of different levels is amended in regard to production sharing agreements: 60 per cent of proceeds will be transferred to the federal budget instead of 60 %, and relatively 40 per cent to the regional budgets, instead of 20 % from income tax.
The regulation, defining the tax rate, established by RF CB for taxes, paid in foreign currency, as of the date of tax payment is transferred to the basic part of the Tax Code (Article 45) from Part 2.
Benefits on VAT and excise taxes on goods imported to the territory of Magadan special economic zone are suspended.
The rules governing the procedure of debt restructuring in the arrears for the insurance contributions of public institutions to the budgets of public extra-budgetary funds, as well as the schedule thereof (the insurance contributions should be settled before 1.01.2010, penalties from 1.01.2010 to 1.01.2020, etc.) are stipulated in the Tax Code.
14. Federal Law N 208-FZ of 24 Nov., 2008 has amended Art. 346.14 of the Tax Code. In a simplified tax system the taxpayers are allowed to change the subject of taxation annually, rather than once in 3 years. To do so, they should notify the tax authority before December 20 of the year preceding the year, in which the taxpayer is going to change the subject of taxation. This provision is in the effect since 1.01.2008.
15. Federal Law № 209-FZ of 24.11.2008 has revised the list of organizations, whose sales of products are not subject to VAT taxation. In addition to Public Unitary Enterprises in the antiphthisic, psychoneurological, psychiatrical and other institutions of social security and rehabilitation, the workshops in criminal-executive system are enclosed.
16. Federal Law № 224-FZ of 26.11.2008 has introduced amendments to the basic and special parts of the Tax Code. In particular, the procedure is provided for postponement and tax payment by installments within five years, granted directly by the Minister of Finance (when the debt exceeds RUR 10 billion and a compulsory debt redemption would lead to the bankruptcy of organization).
Amendments are made to the procedure of suspending bank operations with the accounts of the taxpayers, desk audit procedure; the amount of penalties is increased for late submission of information by banks or failure to provide information on opening and services for the accounts of notaries, individual entrepreneurs, lawyers, etc. (from RUR 10 to 20 thousand and from RUR 20 to 40 thousand, respectively), etc.
Buyers, purchasing discarded property of public institutions are regarded as tax agents.
The Tax Code provisions on the procedure of VAT payment and refund is revised.
Income tax deduction amount for the purchase of housing is increased from 1 to 2 million rubles.Pages: | 1 | ... | 9 | 10 | 11 | 12 | 13 | ... | 15 | Книги по разным темам


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About Me
I am working full time as a Light Worker, Spiritual Teacher, and Messenger for Creator, Angels, Mother Earth, and Healing Energies. I love my job in helping people open up to their Higher Selves. Every one is a Divine Being, all Perfect, and sometimes we just need to be pointed or helped onto our "Path of Divineness".Read more…
Angel Intuitive Practitioner
In these workshops, you will learn to open up your clairvoyance, connect with Angels, and Archangels. Learn how to heal yourself and others using powerful energies that raise your vibrations to heal disease and manifest. You will be certified to give Angel healings, readings, chakra healings, mediumship and channeling...More info…
WorkshopsHealing Workshops, Healing mini Retreats, Vision Quests, Animal Communication... Learn More
Intuitive Readings
Intuitive readings will help give you validation and clarity about your life.Find out more…
The Angel Store
Angel sprays, salts, oils, Angel cards, jewelry, and more!Let's shop…
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Keep Current
Stay up to date with all of Susan's events.View Event Calendar


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March 22, 2016
Islamic Terrorists have claimed responsibility for a large-scale coordinated attack in Brussels today, killing 34 people and critically injuring at least another 200 people. ISIS terrorists struck the city's main international airport and in a subway station at the heart of the city, near the headquarters of the European Union.
The attacks started with a large explosion at a departure terminal at the Brussels airport. Witnesses described an apocalyptic scene of carnage, after an ISIS terrorist set off a suicide bomb and turned the terminal into a war zone.
79 minutes later a second explosion ripped through a Metro station just 400 meters from the EU headquarters, killing at least 20 more people and injuring a countless number of bystanders.
ISIS Claims Responsibility
In the afternoon, Amaq, a news agency affiliated with the Islamic State, issued a bulletin…This post was originally published on this site


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Welcome to the Guidance Department
The mission of the Pope John XXIII High School Guidance Department is to foster an environment in which all students are encouraged to become lifelong learners. By working with the same group of individuals over a four-year period, the counselors strive to nurture the healthy development of all students, acknowledging their individual and cultural uniqueness. The counselors work proactively to develop collaborative relationships with their counselees, their families and staff.
To address students' needs, guidance counselors provide the following services:
- Academic planning
- Course scheduling
- Develop a 4 year plan with students
- Monitor academic progress
- Provide counseling services and make appropriate referrals
- Evaluate promotion and graduation eligibility
- Provide college and career counseling
- Coordinate the college application process
- Inform students of educational opportunities


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Prior to recorded history, a small genetic mutation occurred that created a race of humans now found among every civilization in every corner of the globe. Unrecognizable to the rest of mankind but living among us and dependent upon humans for survival, this race is known as Mortui; those whose spirits do not sever from their bodies when they die. They are the source of all living dead legends.
At the time of his premature death, Wesley Lohmann discovers he belongs to this select group.Wes' transition plunges him into the center of a struggle between those who seek to coexist peacefully with humans and those who wish to use and rule them, a struggle that carries over into the treacherous world of dreams. The delicate balance human and post-human societies have maintained for millennia is threatened by a recent scientific discovery that offers Mortui the hope of returning to human life, and human sensation, but at a terrible cost. As he struggles with new and past relationships and tries to find meaning for his continued existence, Wes discovers within himself the true meaning of being alive.
AfterLife transcends the typical zombie or vampire genre and audience to present a plausible explanation of where such stories come from in an unsettling but believable way. Even those who usually shy away from this type of science fiction are falling prey to the "what if"question that makes this book so unique and is attracting readers who typically enjoy other types of fiction.
Tens List:
Ten pieces of advice I would tell the "teen" me:
1. Yellow is not your color. Don't try to pull it off.
2. Don't waste so much time trying to find a haircut you like. By the time you figure it out, you'll be losing your hair and it won't matter anyway.
3. Read for pleasure. Books are fun when you don't have to write essays about them.
4. Learn to get out of your comfort zone from time to time. The earlier you learn to do this the more you'll enjoy life.
5. Take care of your back. Good posture doesn't seem important now but it will effect on you in the future.
6. Learn to play a musical instrument. It's something you're always going to wish you had learned to do and it'll be easier to do while you're still young.
7. The world is not as big as you think it is. The older you get the smaller the world becomes.
8. The "unpopular" crowd outnumbers the "popular"one. Become friends with people who can be your true friends during the rough times in your life. Having few good friends that care about you is more important than having tons of friends that disappear when you need them most.
9. Don't focus so much on what others think of you. It doesn't matter.
10. The sky is blue. Some people might tell you from time to time that it's a different color but you should never believe them.
Get AfterLife FREE TODAY on Amazon here.
Thanks for visiting! This post was also published on my other blog, FanGirl Hostess.


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The 50th Class Reunion is the second weekend in August, 2018. Email email@example.com for details!
We are still looking for email addresses for some of our classmates. If you have any addresses, phone numbers or emails of people, please send them to us and we'll be better able to stay in touch with those classmates. We'd like to have emails as they are the most cost effective, however a phone number or mailing address is very helpful as well. If you don't know what info we have one someone, please login to see. You can email us at firstname.lastname@example.com.
Help others find out about us… please send a link to this website to class of 1968 members who might be interested.


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What was the Best Country to Live in 2010?
If you read my post back in January, Where Should I Live in 2011, you would have learned that the United States was the best country for quality of life in 2010. Looking back over the past year, even over the past few months you can see that a lot has changed in our crazy world. Has it changed enough to dislodge the U.S. for that #1 postion - probably not. In fact I would say recent world events have probably solidified that decision even more. The U.S. is still one of the healthiest and wealthiest countries to call home. Relative to recent international turmoil be it earthquake, Tsunami or revolution the states is also one of the safest and most stable countries on the planet, save perhaps Canada which one most certainly rank up their as one of the best places to live.
Best Places to Live in The World
But lets have a look at what may have changed over the past few months in terms of the Best Places to Live in the World. Canada for one was suspiciously absent from the Top 20 Countries for Quality of Life in 2010. I'm certain that was some kind of oversight or typo as Canada ranked #9 on the 2009 list. Given the rising cost of oil and the strength of the Canadian dollar I've got to believe Canada would now move quite close to the top of the list. If market indicators are any measure it may very well be number as the U.S. dollar has slipped to a value of 95 cents compared to the Canadian Loonie. Japan is another country that we have all been watching closely. Sure it has had soem economic woes over the past few years with a crippling debt and aging population. However, this small island nation still ranked 7th on the 2010 list of best places for quality of life. The recent earthquake followed by a devastating Tsunami and now an ongoing and as yet unresolved nuclear reactor problem will surely have pushed Japan well down the list if not even off the list altogether.
All in all the events of the past few months have certainly confirmed that we are living in challenging and constantly changing times. Lists are fun and interesting to read but current events tell all. For me I'm thinking a peaceful little place like retiring in Belize may just be the solution - it may even bring me right back to finding the cheapest places to live!


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Get Hidaya (guidance) from The Creator and give Hadiyyah (gift) to His creation
2017 Report
2014 Report
The majority of the people Hidaya Foundation helps are poor villagers who live in primitive dwellings such as mud huts without electricity. For such people, having a few animals could be a very good way of sustaining themselves for their own personal food as well as for profit and trade.
Poultry Farming: Five chickens and one rooster, or a flock of 25 chicks, are given to a family (feeding supplies are also provided). It costs about $50 to start a small poultry farm.
Goat Farming: Goats are given to the poor in rural areas for milk and goat farming. It costs about $220 for a pair of goats for a family to get started.
Cow Farming: It costs about $400 for a cow which can provide milk.
Sign up for Automatic Monthly Donation towards this project.
Hidaya Foundation was awarded Sitara-i-Eisaar (Star of Sacrifice) by the Government of Pakistan, in recognition of the results-oriented relief efforts during the 2005 earthquake in Pakistan.
Hidaya Foundation is a non-profit 501(c)(3) with FEIN 77-0502583. All cash and in-kind donations are tax deductible. Hidaya puts it's best effort to utilize the funds donated for a specific project, sometimes circumstances dictate we use funds otherwise.
Warehouse for in-kind drop-off: 57 Bonaventura Dr, San Jose, CA 95134 (Open Sat & Sun 12 - 4pm)
Mail Donations to: Hidaya Foundation, PO Box 5481, Santa Clara, CA 95056
Tel: 408-244-3282 | Toll Free: 1-866-2HIDAYA | Fax: 1-866-3HIDAYA | E-mail: firstname.lastname@example.com


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NGV Friday Nights returns this winter with exclusive late night access to MoMA at NGV. Featuring a line-up of international and home-grown acts presented by the Melbourne International Jazz Festival, experience art, music and dining inspired by the city that never sleeps.
7–7.45pm & 8.45–9.30pm
Linda May Han Oh first began playing piano and bassoon before picking up the electric bass at fifteen. She went on to study at the W.A. Academy of Performing Arts (WAAPA) before completing her Masters at the Manhattan School of Music in 2008. Linda has performed with musicians such as Joe Lovano, Vijay Iyer, Dave Douglas, Kenny Barron, Geri Allen, and Terri Lyne Carrington. Based in New York with an active international touring schedule, she is currently the bassist with guitarist, Pat Metheny.
Fabian Almazan found his musical roots as a child in his homeland of Havana where he first became involved in the classical piano tradition. After moving to Miami, Fabian studied at the New World School of the Arts High School from 1998 to 2002. Since 2007 Fabian has been the pianist for Terence Blanchard's various bands, including the E-Collective which was nominated for a Grammy in 2016. He has toured North and South America, Asia, Africa, Australia and Europe extensively and has performed with artists such as Linda May Han Oh, Gretchen Parlato, John Hollenbeck, Paquito D'Rivera, Mark Guiliana, Dave Douglass, Chris Dingman, David Sanchez and Ambrose Akinmusire.
Linda and Fabian will perform with performance staff – Rob Burke, Jordan Murray, Paul Williamson and Dave Beck – from The Sir Zelman Cowen School of Music, Monash University.
Angus and Harriett
6–7pm and 7.45–8.45pm
Angus Gray is an instrumentalist, vocalist, composer, producer and arranger based in Melbourne. His piano skills are highly sought after on the jazz, funk, and blues music scene, and he is a recipient of the Will Poskitt Piano Scholarship and Lionel Gell Foundation Scholarship for Jazz and Improvisation at The University of Melbourne. Harriett Allcroft is an Australian vocalist and composer, known for her mesmerising yet light-hearted style. Originally from Adelaide and now based in Melbourne, Harriett performs original contemporary jazz and re-arrangements of Australian jazz tunes with her trademark playfulness and whimsy.


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Ara Chackerian may be commonly known by the public to be a successful entrepreneur, investor and philanthropist, but the fact of the matter is that he is more than that. He is more than just a committed creator of solutions in the medical industry. He is more than just a competent investor of capital in ventures that push for finding alternative ways to treat the kinds of depression that resist treatment.
In the interview that Ara Chackerian did for Ideamensch, it's revealed that most of his time today is spent in developing TMS Health Solutions. Before his venture in this area, Chackerian used to be spending most of his precious time in investment programs and entrepreneurial pursuits. It was when he got initially inspired by new methods of curing diseases that he realized he wants to be actively involved in building healthcare companies. Right now, he has a lot of these companies under his name, which include BMC Diagnostics, Embion/Provider Links and the most recent one is TMS Health Solutions.
Another advocacy of Chackerian today is in promoting different mental health advocacies. He even recently wrote a Medium article sharing his views on what to do during this Mental Health Month. It is an important part of his career to be involved in treating depression, and mental health is nothing, if not a celebration of the ways that depression has now recently been treated. He believes that for this Mental Health Month to be effective, lectures should inform and educate the public on the various efforts needed to prevent one from being mentally unhealthy.
He also added some of the important tips to find out whether one or the loved ones are suffering from depression. Many people die from mental illness and that's why it is important for families and friends to understand the best way to help those who are in need of help in terms of mental health treatment or just diagnosis. Visit his Vimeo account for more.
About Ara Chackerian
Ara Chackerian is well-known for being the Founder of TMS Health Solutions. He went to the Florida State with a degree in B.S. Marketing. You can follow their Twitter page.
Click here: https://vimeo.com/arachackerian


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You can prevent your child from being bullied. The solution to bullying lies in self-confidence for both parties.
According to the National Association of Learning Disabilities: "The average bullying episode lasts only 37 seconds."
However, from my experience, it doesn't take 37 seconds to say, "You're gay."
Or "You suck."
Or "You're an idiot."
It only takes a second for a word to hit like a fist.
Then, if the bully sees from your face that he has connected; he wins and you're marked as a victim.
Patricia McDougall wrote an article for Education.com, What happens over time to those who bully and those who are victimized?
"Researchers have found that bullying roles (those who bully and their victims) remain fairly stable throughout school.
For example, even after switching to a new classroom, victims of bullying continued to be victims."
Children who are overweight, wear glasses, have a learning disability or are simply insecure will often remain victims because they believe the taunts and feel shame over it.
Ben Franklin once observed, "The sting in any rebuke is the truth."
I know.
I was bullied in fourth and fifth grades by my classmates calling me "Baby Bobby."
The element of truth was that when I was a first grader I cried a lot in class.
The name-calling connected with me because I still felt shame about it.
Making it worse, I believed there was nothing I could do to change my victim status in my classmates' memory.
As a result, I was a bully magnet.
It was as if I had a permanent "Kick Me" sign taped to my back.
Nevertheless, despite my history with bullies, I have empathy for them as they are not bullies because they want to be (and I'm only talking about children here).
Like everyone else, they want to feel important.
They want to be cool and popular.
They want to be liked and have friends.
Unfortunately, they have learned that they can gain that sense of importance by bullying.
Their bad behaviour has been rewarded by their peers, so they continue to refine what is working for them.
We need to give them another way to achieve these ends.
Most likely they learned to bully at home – where they are the victims of a sibling or of their parents.
When they pay it forward, their own victimhood becomes less painful or debilitating.
The degree to which someone is a bully, a control freak, narcissist or sociopath is the degree to which they have been abused.
The more severely abused a person was – the bigger the bully he or she becomes.
To see examples of this we can look at Adolph Hitler, Joseph Stalin and Saddam Hussein, all of whom were severely abused as children, and in turn grew up to be tyrants.
People who seek power through bullying do so because they feel powerless.
Confident people don't need power over others.
They have learned to acquire the things they need through peaceful communication.
The time to rescue a bully is when he or she is a child. When they can still learn more positive ways to gain a sense of importance.
Bullies and their victims are opposite sides of the same coin.
The problem for both of them is essentially the same.
They both crave positive recognition, but they both lack self-confidence and the social skills to acquire what they need.
The solution is twofold: helping the victim develop self-confidence, and helping the bully develop self-esteem.
As a society we can do more by giving them what they need.
Previously in this column, I have written about self-efficacy which is the belief that we can achieve our goals.
I have found that the more a child becomes good at something – and it can be just about anything – the more their overall self-confidence grows.
The more visible their achievements are to their peers, the better, because kids admire accomplishments.
It gives them "cred."
One of the ways in which we acquire self-efficacy is from the encouragement of others.
Teachers, parents and other adults who work with children can do so much to make this happen.
Your praise is more powerful than you know; it fortifies victims, and nourishes those who would otherwise become bullies.
– Robert Wilson
• Robert Wilson is an author, humorist and innovation consultant. He is also the author of the children's book, The Annoying Ghost Kid and the recently-published Wisdom in the Weirdest Places. For info, visit www.jumpstartyourmeeting.com.Posted: Sep 1, 2015


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HP CEO Meg Whitman on Integrity & Courage in Leadership
"You have to be known for high integrity – doing what you say, saying what you mean," emphasized HP Chairman, President, & CEO Meg Whitman. During her View From The Top talk at Stanford GSB on April 6, 2015, Whitman shared leadership lessons and offered career advice. Read more takeaways: http://stanford.io/1y8McHC
Source  :  Stanford Graduate School of Business
Category: Fascinating Stories


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Who are we?
The automobile industry is undergoing a profound transformation. New technologies like automonmous driving and advanced data connectivity are redefining the way cars are sold and operated. As a result, our industry needs salespeople who are up-to-speed on the latest technologies, eager to help customers find the best solution for their transportation needs, and excited to be part of the future of transportation.
No company is better positioned for this new world than Holman Automotive Group. Much more than a network of dealerships, Holman has built a 90-year tradition of innovation and success by drawing on our most valuable resource…our employees. Now into our third generation of family leadership, Holman is looking for a new generation of intelligent, motivated and sophisticated sales consultants ready help us meet tomorrow's challenges. If you'd like to work in this kind of exciting, technology-driven, customer-centered environment then Apply Today!
Kuni BMW has an outstanding opportunity for a Client Advisor to join the Holman family.
What will you do?
What are we looking for?


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You can have fun with locks
I read about this guy who made an interesting locking box for a wedding present. It is a locked box that will only open if is in a certain spot on the Earth. It has a built in GPS and when you press the button it will tell you on the screen how far away you are from the location of it opening. It also only allows you to press the button 50 times. If you would like to read more about it here is a link to the guys blog.


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Circuitry is one of my favorite explorations with kids. The excitement when that bulb lights up? There is nothing like it. With basic materials, kids can really start creating their own projects and in the process, learn a lot along the way. A simple entry point is Squishy Circuits, but more advanced projects like adding in motors and other components lead to endless possibilities. You don't need expensive kits like LittleBits or Snap Circuits. While those tools are fun, they aren't a must have. In fact, kids can learn just as much with low-tech materials listed below that can be purchased in bulk. Partner with cardboard, duct tape, and good old paper? And the possibilities just became awesome.
Squishy Circuits
Pro-tip: Playdoh is highly conductive and comes in LOTS of colors.
- Squish the Boredom and Power Up the Play
- St. Thomas Project Page
- Tinkering Studio How To
- Squishy Circuits Classroom Guide
- Gadgiteration Guide
Paper Circuits
- The Tinkering Studio: Paper Circuitry
- HighLowTech
- Instructables for Beginners
- Paper Circuits from Makerspaces.com
Copper Tape & Conductive Inks
- Copper Adhesive Tape [Note: Look for copper tape with conductive adhesive!]
- How to Make Conductive Ink
- CiruitScribe Conductive Ink Pens
LED Lights
- 10mm are fun for tiny hands, but pricier
- 5mm bulk LED's in multiple colors
- Chibitronics Stickers
- 3V Button Cell, packs of 100 on Amazon are the best deal.
- Ikea Button Cell is a good deal in store
- Battery Holders


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Create Content in MinutesGet Started For Free
Craftsy asked its members: "Should art ever be censored?" people have multiple opinions wether art should be censored or not. This relates to speech because wether or not, this still is an act of wether you have a say in art.
A man speaks whats on his mind and starts mayhem for some people, causing the president even to get in the act. Even though some people may not be right, freedom of speech should be even given to this individual.
What is FREEDOM OF SPEECH? What does FREEDOM OF SPEECH mean? This articles gives you a brief description of what freedom of speech is.
A professor at Yale encourages students what they should wear on Halloween, causing the students to make rules on what to wear and using offensive costumes.


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1. The Committee has considered the instruments set
out in the Annex to this Report and has determined that the special
attention of both Houses does not require to be drawn to any of
2. The Committee draws the special attention of
both Houses to these Regulations on the ground that they make
an unexpectedly limited use of the enabling power.
3. Regulation 53(1) creates a right of appeal to
the Secretary of State or the Food Standards Agency on the ground
that the amount of the charge paid by a person for veterinary
checks to a local authority is unreasonable. If the appeal is
successful, regulation 53(2) requires the local authority to recalculate
the charge and repay the difference. Regulation 53(3) creates
a right of appeal to an independent person on the ground that
the amount of the charge paid to the Secretary of State or the
Agency is unreasonable. However, no provision (corresponding to
regulation 53(2)) is made in this case requiring the recipient
of the charge to repay the difference if the appeal is successful.
The Committee, in its Eleventh Report (Session 1996-97)), reported
earlier Regulations (S.I. 1996/3124) on the ground that the
absence of such a provision constituted an unexpectedly limited
exercise of the enabling power. In a memorandum printed at Appendix
1, the Department for Environment, Food and Rural Affairs accepts
that a provision should be included in regulation 53 for recalculation
of the charge and repayment of the excess where the amount of
a charge paid to the Secretary of State or the Agency is held,
on appeal, to be unreasonable. It undertakes to include such a
provision at the first available opportunity. The Committee
accordingly reports regulation 53 on the ground that it makes
an unexpectedly limited exercise of the enabling power.
4. The Committee draws the special attention of
both Houses to these Regulations on the ground that they are defectively
5. Regulation 3(2) provides for the functions of
the children's rights director set out in regulation 3(1)(b) to
be subject to the requirements of the Care Standards Act 2000
and section 87 of the Children Act 1989. It was not clear to the
Committee what requirements of the 2000 Act were contemplated
and why it was necessary to make this saving for that Act and
section 87 of the 1989 Act. In a memorandum printed at Appendix
2, the Department of Health explains that the contemplated provisions
of the 2000 Act are sections 13, 31,32, 45 and 46, and that the
saving was considered necessary so as to avoid any possible inconsistency
between these provisions and the procedure for dealing with applications
for registration, and the methodology for inspection of premises,
to be established by the director. In relation to the saving for
section 87 of the 1989 Act the Department states that this reference
was included so as to ensure that the director exercises his function
consistently with the powers of inspection provided by that section.
In the Committee's view, the saving in regulation 3(2) for provisions
of primary legislation is both unnecessary and misleading. It
is unnecessary because secondary legislation is subordinate to
primary legislation; and it is misleading in suggesting that,
without the saving, the legal position might be otherwise. The
Committee accordingly reports regulation 3(2) for defective drafting.


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Inhalation therapy is an area of research focus at Bend Research. Our pulmonary drug delivery and nasal drug delivery technologies leverage more than 15 years' experience in formulation development and particle engineering.
We have expertise in:
- Particle engineering
- Development of novel inhalation dispersion excipients
- Formulation and particle embodiments
- Equipment and scale-up
- Dry powder formulation development for biotherapeutics
- Capsule fill for dry powder aerosols
Bend Research provides feasibility and formulation identification for dry powder compositions of small molecules and biotherapeutics. We have a full scale-up train from mg scale to kg scale, cGMP manufacturing facilities, and capsule filling capabilities. Analytical, chemistry manufacture and control, stability, and in vivo studies are supported through our collaborator network.
Particle Engineering Experience
At Bend Research, we have the equipment, process and formulation know-how to develop engineered particles across a wide range of sizes and size distributions for a wide variety of delivery routes and applications. All approaches have been demonstrated as feasible through current good manufacturing practice (cGMP) manufacture.
Development of Novel Inhalation Dispersion Excipients
A critical factor in inhalation dispersion platform technology is the identification of safe excipients with high glass-transition temperatures (Tgs). Dextran and dextran-based materials provide an overlay of performance and precedented use in clinical studies. Using this approach, we have successfully formulated many different types of compounds including combination therapies, biotherapeutics and small molecules.
Formulation and Particle Embodiments
Lack of precedented excipients for inhalation and interest in combination therapies has led Bend Research to pursue multiple creative formulation concepts. All terminally utilize the spray-drying process to generate respirable powders.
Equipment and Scale-up
The spray-drying process leads to a scalable way to tune particle size to obtain highly efficient dry-powder aerosols. Bend Research has custom equipment designed to efficiently manufacture and collect particles of sizes and size distributions suitable for inhalation. The scale-up train delivers consistent product from material-sparing feasibility studies (milligram quantities) through large-scale manufacture (several kilograms) of clinical trial material.
Dry powder formulation development for biotherapeutics
There has been increasing interest in delivering biotherapeutics to the lung. Very often, a dry powder is ultimately desired and current lactose-based technologies are not appropriate. Using approved, precedented or novel excipients, Bend Research identifies formulations and a gentle spray-dry process that leads to stable dry powders with excellent aerosol performance. Our expertise in spray drying and custom equipment enables rapid material sparing formulation identification that can then easily be scaled to our GMP facility for toxicology supplies or clinical studies.
Capsule fill for dry powder aerosols
Very often in early proof-of-concept studies, it is desirable to rapidly begin clinical studies with a simple and commercially available
capsule-based dry powder inhaler. To support this, Bend Research has dry-powder capsule-fill capabilities to support stability, toxicology studies and Phase 1 clinical studies.
Download the PDF presentation below for an overview of our inhalation technology:


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Those who care deeply about pizza probably know about Liz Barrett. Liz is editor at large for PMQ Pizza Magazine, and blogger at The Pizza Insider. She's just released her first book, Pizza: A Slice of American History. As soon as I could get a copy, I did - and I devoured it almost as fast as a DeLorenzo's tomato pie. Click HERE to find her book on Amazon.
I appreciate when a book is well-written and thoughtfully organized; I think of her pizza book as the necessary "Volume II" that follows Ed Levine's Pizza: A Slice of Heaven. Read them both for a valuable perspective on pizzas new and old.
I had some questions for Liz after reading her book; here are her answers.
PQ: Liz, in your book you quote Ed Levine, who says that the first pizza of your childhood becomes the metric against which you judge all pizzas. Tell us about your first pizza, and if it still holds a place in your heart even though you now know of so many world-class pizza styles and pizza makers.
LB: My first pizza was in Troy, Michigan - typical Detroit-style pizza with the thick crust, crisp bottom and caramelized cheese around the edges. No matter how many pizzas I eat, I still get excited when I hear that a place I'm visiting may have an authentic Detroit-style pie.
PQ: I particularly enjoyed your chapter on the tomato pie, the Trenton NJ version of pizza, because that was my first pie and the inspiration for my blog. Despite having eaten (or searched) for Trenton-style pie my entire life, I never heard the term "reverse pizza" applied to tomato pie, as noted in your book. Can you tell us where you heard that, and also your own tomato pie experiences?
LB: That's a good question. I've interviewed so many people over the years and talked with so many pizza experts that I don't know if I could pinpoint the person who called it that first. I'd hate to name the wrong person here. It makes sense though; the pizza is assembled in a reverse fashion, with cheese on the bottom and sauce on top.
PQ: One aspect that elevated your book beyond the mere passion of a pizza fan was your thoroughly researched approach. I dug deep into the chapters on Detroit style and St. Louis style pizza, because they sound (and look) wonderful and I've yet to try either. Have you sampled all the styles in your book? What kinds of pizza remain on your "must try that" list?
LB: I'm happy to say that I have tried all of the main styles in the book. There are only a few styles in the More Pizza Styles chapter that I have yet to try, such as grilled pizza, Quad City-style pizza and Colorado style mountain pie.
PQ: I've noticed a lot of overlap in pizza styles. Your book covers the bar/tavern/party cut thin-crust pies, with the wonderful Rubino's (Columbus, OH) as an example. I often think that a New Haven apizza has much in common with a Trenton tomato pie. And a Philly tomato pie is, to me, a Sicilian pizza with no cheese. Are there other hybrids or overlaps that stand out from your research or pizzeria visits?
LB: You're right; there's some crossover with pizza styles, mainly because of the close proximity to other towns and the fact that pizza makers move from one place to another and integrate their skills into the next place. Many people have compared Detroit-style pizza to square pan pizzas and Sicilian pizza, but without the caramelized cheese on the edges, you don't have Detroit style, in my opinion. I don't think we've seen all the pizza styles we're capable of yet.
PQ: My blog's tagline is "La pizza male è meglio che non la pizza" -- even bad pizza is better than no pizza. But let's be honest - some pizzas are better than others. Your books offers a passionate yet fair-minded analysis of all the pizza styles - but is there a type of pizza that you don't like? For instance, I have never had Old Forge style, even though it wouldn't be a long trip from my home to try it. From pictures, it looks like a soft, gloppy, badly undercooked attempt at Sicilian pizza. Is there any pizza style you judge to be unworthy of the calories?
LB: There are some pizzas I haven't finished; but I always give them a chance (it's a tough job, but you know).
PQ: While on the subject of bad pizza, let's talk about the chains. As your book notes, the chains are very much responsible for the popularity of pizza in America. Without the chains making pizza into a cheap, tasty, and convenient dining option, we probably don't arrive at the Neapolitan pizza renaissance taking place right now all across the country. Nonetheless, I feel that most big chain pizza can't compete with a good frozen pizza. However, I find a lot of the smaller chains are making some good to excellent pies. Grottto, Jules Thin Crust, Monical's, Bertucci's, and most of all Anthony's Coal-Fired Pizza. Do you have a favorite chain pie, big or small?
LB: There are times when I just need some delivery. And much like your quote above, I'd rather call for some pizza than have no pizza at all. My delivery pizza of choice is usually Domino's. I like the seasoned crust they added a couple of years back.
PQ: Lastly, do you have a guilty pizza pleasure? A particular pizza, or topping, or other decadent pizza habit? I admit, I will wolf down a monstrous slice of $2 Costco pizza a little too often. I know it's not "good" pizza but I enjoy it anyhow. Anything you'd like to confess?
LB: While it goes against everything traditionalists fight for, I can't resist a barbecue chicken pizza. Just love it.
|Inside Liz Barrett Foster's Pizza Book
PQ: Liz, thanks for taking the time to share your thoughts with Pizza Quixote blog readers, and thanks for that thoroughly-researched tome on pizza. It's added to my growing list of "must try that" pizzas.


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Gru is finally settling down. His new life with Lucy and the girls is the furthest thing from what he had imagined, yet it's exactly what he never knew that he wanted. Just when he thinks he has things figured out, a mysterious stranger shows up to inform him about his long lost brother. To make matters even more difficult, it's his twin brother that he never knew about. The journey of meeting and getting to know Dru (his twin), will re-ignite a passion for villainy. So is Dru enough to cause Gru to relapse into his old ways? Or, will Gru's new life keep him on track? This is a bit unconventional in its presentation but there are references/easter eggs that pay homage to other family reunion type of films. A heartwarming adventure that is safe for all ages, it's specifically aimed at a younger audience but fans of the other Despicable movies will find that there is more to enjoy. Unlike the others, each of the characters has a few of their own stories along the way, some are cute, some are corny but they're all fun. Is it enough? Only you can decide.
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