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RNA interference

The Nobel Prize in Physiology or Medicine in 2006 was awarded to Andrew Z. Fire and Craig C. Mello for their research on RNA interference. The goal of this learning project is to complement the Wikipedia article about RNA interference in two ways. The first goal is to provide a user-friendly introduction to the topic. This means providing learning resources for people who would normally be unable to understand a technical Wikipedia article on the topic of RNA interference. The second goal is to provide learning resources that allow interested university students to collaboratively explore the science behind each awarded Nobel Prize in more detail than is possible with the related Wikipedia article. If you have not done so already, take a look at the Wikipedia article about RNA interference then select one of these learning paths:

 * 1) Explore a user-friendly introduction to the practical medical implications of RNA interference that arise from the Nobel Prize-winning scientific research of Andrew Z. Fire and Craig C. Mello.
 * 2) If you were able to read and appreciate the Wikipedia article about RNA interference then continue reading below and participate in further exploration of this subject.

RNA interference was discovered as a mechanism used by cells for regulating gene expression. This discovery has quickly resulted in the widespread use of artificial interfering RNAs as an important laboratory research technique for altering the amount of specific proteins inside cells. There is also active study of the potential value of RNA interference for medical applications.

Short review: RNA interference in cells

This section and the next few sections briefly introduce RNA interference (RNAi) and will orient you towards the activities. See the Wikipedia article about RNA interference for more a more detailed introduction to RNAi.

The normal function of RNA interference inside cells depends on the production of double stranded RNA (dsRNA). Base pair-complementary RNA strands (ssRNA) can be produced by transcription of both template DNA strands of some genes (Figure 1). In other cases, the protein-coding RNA sense strand might be produced by a virus and the antisense RNA strand produced by the host cell. The sense and antisense RNA strands form double strand RNA (Figure 2, top) that is processed to small (about 20 base pairs long) inhibitory RNA (siRNA). The siRNA can form a molecular complex with proteins that first strip away the sense strand of RNA, making the antisense inhibitory RNA (iRNA) available for base pairing with messenger RNA (mRNA). This targets a specific mRNA for destruction (Figure 3), resulting in inhibition of the biological function that would otherwise be served by the protein coded for by the mRNA.

There had long been interest in the idea that it might be possible to alter gene expression by using single strands of nucleic acid that might base pair hybridize with a specific target in cells. However, it was only in 1998 that experiments were described showing the unexpected power of double stranded RNA to block gene expression. The experimental system used was the worm Caenorhabditis elegans. This was a useful experimental system because the developmental orgin of all of this organism's cells is known and it is possible to inject RNA into early embryos and observe changes in the pattern of development.

Protection against viruses
An important aspect of RNA interference is its role in protecting organisms from some deleterious effects of viruses. Such a role for RNA interference was first found in plants, but has also been found in some animals.

Gene expression knockdown experiments
RNA interference is now a widely used biology research technique that can be applied to both cultured cells and whole animals. RNA interference can be used to selectively reduce the level of expression of a specific protein. Clues to the function of the protein can be obtained by observing changes in cell or organism behavior after knockdown of expression.

Medical applications
It has been suggested that knockdown of expression of proteins by virus-delivered siRNA might be effective for future treatments of Amyotrophic lateral sclerosis  and other neurodegenerative diseases. Working with laboratory mice as an experimental model system for the human disease ALS, Miller et al showed that loss of muscle function could be slowed using RNA interference.

Detailed Review
RNAi is discovered as a defense system to prevent attack of viral and parasitic RNA. Later on, other fuctions of its emerged, like endogeneous gene regulation. This system is able to seen in Caenorhabditis elegans, Drosophila, yeast as well as can be done in mammals. Main mechanism of RNAi is revealed by Fire and Melo. It is used in many laboratories to induce gene silencing in organisms by delivering appropriate dsRNA (double stranded RNA). RNAi can be utilized as an analysis tool to understand function of genes therefore let us to obtain an evolutionary view. It is also possible to determine mutant genes, which cause death of the organism, via RNAi. RNAi centered techniques for full genomic screen provides oppurtunity to identify specific genes via testing a given trait with high trustability. CRISPR interference is an alternative for RNAi in prokaryotes. CRISPR and RNA interference mainly differ at structures of the components, however, pathways share similarities. Increased studies ulliminated the way that RNAi work and let scientist to utlize it better.

Basic Mechanism of RNAi in Mammals
Three main components of RNAi silensing includes Drosha, Dicer and Argonaute (Ago) proteins. Both Drosha and Dicer are the members of RNAase III family. They have ability to cut dsRNA by leaving 5’ phosphate head and 2 nt (nucleotide) overhang at 3’ end. With this property, they are responsible for processing of newly transcribed small hairpin non-coding RNAs to smaller pieces. The pieces are 21- 25 nt dsRNAs. Drosha only found in animal cells. After, one of the strands of 21- 25 nt dsRNAs loaded into RISC (RNA induced silensing complex). This complex is the direct effector of mRNA translational inhibition and it is mainly based on members of Argonaute (Ago) protein family. Slicing activity of Ago is dependent to perfect complementarity of guide RNA with target RNA. Imperfectly matching leads to generation of several scenario containing translational inhibition, prevention of de- adenylation and nuclease degradation by P body. While each scenerio is possible alone itself, also combination of them can be observed. Some Ago including complexes may provide inhibition of transcripition via affecting chromatin structure or causing modification on histone proteins .Basic mechanism of post-transcriptional gene silencing in mammals is given step by step in figures named Part 1, Part 2, Part 3, Part 4, Part 5.

RNAi in other Organisms
RNA interference is an highly conserved mechanism but components of the system may be distinct from one organism to another, even some contributers of the mechanism may be deficient in some cases.

C. elegans: C. elegans is a transparent roundworm. It has 1mm length. Diet of its consists of bacteria. By means of the efforts of Dr. Sydney Brenner, this animal became one of the strongest model organism. Ago- guide RNA interraction forms the base of RISC(RNA induced silencing complex). Ago has two extremely characteristic domain called PAZ(PIWI/ Ago/ Zwille) and Piwi. These domains are responsible for holding guide RNA in a way that does not close the side which base pairing occurs with target mRNA. In PIWI, catalytic role is borne by DDH motif which is similar to RNase H family catalytic. Problem on the residues of DDH on Ago2 results in cancellation of slicing of target.

RDE- 1 is a type of Ago in C. elegans, works with rde- 4. Without rde-4, decline on siRNA level happens. RDE- 4 has a binding motif which provides better interraction with long dsRNA, instead of short ones like siRNA. This protein is not just linked with rde- 1 and dsRNA, also have connection with the other elements of RNAi. Rde- 4’ s main responsibility is recognition of long dsRNA and presentation of it to DCR- 1.

SID- 1, a type of membrane protein of C. elegans, provides passage of exogeneous dsRNA, into C. elegans cytoplasm. Homologs of SID- 1 found in mammals however not found in Drosophila. MicroRNA has crucial effects on differentiation and time of development of this organism. Different organisms have different anatomies and physiologies which cause application of separate techniques to force organism to accept service of dsRNA. Introduction of dsRNA to C. elegans can be achieved via several methods. These methods are:

a)	Direct injection. dsRNA including solution is ejected to the body cavity. In this method, RNAi response arises throughout the body.

b)	Production of trangenic amimal that express target dsRNA. The technique is limited with the tissue that the technique applied.

c)	Immersion of the organism to the solution of dsRNA. Ingestion of RNAs leads generation of RNAi response.

d)	Feeeding with bacteria which are genetically modified to produce target dsRNA.

Drosophila: Drosophila genus hosts one of the model organism named Drosophila melanogaster.

In Drosophila, dicer- 1 and dicer- 2 found as paralogs of Dicer. As dicer- 1 plays role in generation of miRNAs, dicer- 2 included in synthesis of siRNAs. Explanation of this discrimination originates from structural differences. While dcr- 1 does not have helicase domain, dcr-2 suffers from absence of PAZ domain. However, in terms of function, dicer- 1 and dicer- 2 don’ t have a thick boundary wall that separates them from each other. Both of the enzymes are obligatory for functioning of the siRNA mediated destruction of target mRNA and gene silencing. PAZ domain of Dicer works with RNase III domains in order to provide determination of 3’ overhang ends of target RNAs. RNase IIIa and RNase IIIb gather to take over catalytic process that dicer is responsible for. Linkage of these domains with Ago occurs at the side of PIWI domain of Ago. Efficiency of the enzyme is highly dependent to 3’ overhang nucleotide sequence After Dicer processing, small noncoding RNAs passed into RISC.

List of members belongs to RISC of Drosophila  :
 * Ago2
 * dFXR (Drosophila Ortholog of Fragile X Mental Retardation Protein)
 * VIG (Vasa Intronic Gene)
 * Tudor- SN
 * R2D2
 * Aubergine
 * Armitage- RNA helicase
 * Others

While RISC loading occurs, guide strand selection is carried out by the help of R2D2 protein. As passenger strand is gong to be degraded, other one used as a guide. Identification of passenger and guide strand of siRNA is achieved accourding to thermostability of ends of strands. At the time of guide strand selection, Dicer connects to less stable 5’ end of siRNA as R2D2 binds to more stable one, in thermodynamicity. Less stable ended one loaded into RISC. In Drosophila, separation of passenger and guide strand is carried out by Ago2.

Usage of less stable strand on RISC lowers, even minimize, posibility of off targeting. PAZ of Ago contact with 3’ end of siRNA while PIWI domain makes connection both with RNase III domains of Dicer and phosphate headed end of siRNA single strand. PAZ affinity towards 3’ overhangs originated from aromatic residues on it. Different members of Ago family bind to distinct siRNAs. All of four Ago of mammalian cells can connect to miRNA. As opposite to mammals, Drosophila melanogaster has only Ago1 in its miRNA pathway.

Drosophila has 3 type of siRNA based RISC as shown in table located below.

The I Factor is the structure that is the cause of the disorder named as Hybrid Dysgenesis, in Drosophila melanogaster. This disorder contains abnormalities mainly based on high mutation rate. I Factor repression is maternal and have long term effect. Repression of the factor carried out by the help of RNAi. One strong scenerio about it suggests siRNAs for repression originated from egg cytoplasm. Another elements occurs to cause other types of Hybrid Dysgenesis. One of them is called P element, which are DNA transposons. High level of transposible element activity drives cell to death. P element is under the control of RNA mediated mechanism.

PEV (Position Effect Variegation) is the result of chromosomal rearrangement of the place of euchromatic gene. The euchromatic gene transferred next to heterochromatin and euchromatic characteristics of the translocated gene disappears, gene become silenced. PEV is well studied in D. melaogaster eye and body pigments. PEV is mainly related with propagation of methylation.

HP1 based silencing is achieved by the help of RNAi components. Expression of transposable elements of Drosophila is majorly controlled at transcription level. RNAi connected to gene silencing via Polycomp proteins in Drosophila. Polycomb response elements helps silencing via employing Polycomb complex. Stuck at RNAi machinary provoke rise in transcription of heterochromatic genes.

S. pombe: Although Schizosaccharomyces pombe is a fungus, because of its high similarities with other eukaryotes, it became a good choice to work on some cellular events including RNAi based heterochromatin generation. In S. pombe, also known as fission yeast, RNAi has considerable effects on the formation of heterochromatin. Centromers, telomeres, locus of mating- type and ribosomal DNA are the regions under the control of RNAi mechanism in terms of chromatin modification.

Heterochromatin is the name of sites exposured to methylation on lysine 9 of H3, a type of histone protein. Methylation occurs with the help of Clr4, a histone methyltransferase. Number of methyl inserted to H3 can vary from one to three. Loss of Clr4 directly means that loss of H3K9 methylation. Clr4 has chromo and SET domain. SET plays role in the methylation. Abnormality on SET domain causes results in changes on the methylation level at some loci of S. pombe.

Clr4 also mediates Swi 6 to bind to related loci. Methyl addition to lysine 9 residue lets Swi 6, a chromo domain protein in fission yeast, to link to H3 by the help of its chromo domain. Dimerization of Swi 6 proteins may lead other proteins to associate with Swi 6 via interraction surface resulted from dimerization.

Acetylation of target residue blocks methyl insertion. Therefore histone deacetylase activity is needed. This requirement is carried out by the enzymes of HDACs (histone deacetylases). Sir2 and Clr3 are the examples of HDACs with Clr6, which has broad target catalog that makes it crucial.

Presence of RNAi mechanism is as significant as functioning of Clr4 and Swi6. RITS (RNA induced transcriptional silencing complex) and RDRC (RNA directed RNA polymerase complex) are the two complexes that are the components RNAi mediated generation of heterochromatin in Schizosaccharomyces pombe. RITS composes of Ago1, Chp1 and Tas3 besides siRNA bound to Ago1. In RITS, Chp1 is responsible for interraction of the RITS with H3K9me2 (2 methyl added to lysine 9 of H3) via its chromo domain. In the absence of one or more components of RITS, remainings can associate but activities of this “half” RITS is not capable of doing activities of fully equiped RITS. Dcr1 and Clr4 are required, alongside Rdp1 (RNA dependent RNA polymerase 1) catalytic activity, for RDRC- RITS linkage. Relation of RITS with both siRNA biogenesis and chromatin structural changes is an evidence of presence of feedback mechanism working between RNAi and chromatin modification. H3K9 methylation at the telomeric, centromeric and mating- type region is not totally cancelled out in the absence of RNAi. This is because of RNAi independent pathways in S. pombe, like Taz1 protein in telomeres. Experiments suggest that other chromo protein Chp2 takes over the job of Chp1 at related loci.

Heterochromatin has impotance on mating- type determination. Abnormalities found in the heterochromation structure of mating- type switching genes, can drive haploid cell of S. pombe to death.

Arabidopsis: In plants, the place for Drosha is filled with the Dicer homolog protein called Dicer- like 1. Plants produce four Dicer Like Proteins, abbreviated as dcl. Four of each bears separate jobs. Processes of pre- miRNAs (precursor miRNAs) are carried out via dcl- 1 while dcl- 2 is responsible for production of siRNA used in defense mechanism against to viruses. Dcl- 3 generates siRNAs that are the part of chromatin modification and silencing at transcription level. The last dcl enzyme named dcl- 4 cares formation of trans- acting siRNAs which are involved in regulation of accumulation mRNAs.

Dcl- 1 is escorted by Hyponatic Leaves1 (HYL1) and Serrate (SE) to function in Arabidopsis. While HYL1 is a dsRNA binding protein , SE is the encoder of C2H2 zinc finger protein, which affects miRNAs’ relation with leaf polarity in Arabidopsis. It is current truth that SE and HYL interracts. Dcl- 2, 3 ,4 are mainly involved in siRNA generation pathway. Dcl- 1 also has crucial missions in the formation of natural antisense short interfering RNAs. Thus, this event is an example demonstrates dcl- 1 is not restricted with miRNA genesis.

Before process of Dcl, Arabidopsis utilizes HASTY, homolog of exportin 5, in order to export miRNA from nucleus.

Methylation of miRNAs is carried out via HEN1 protein as a general step of miRNA biogenesis in plants. Methylation happens after Dcl functions. 2 overhang of 3’ end of miRNA duplex is favored substrate for HEN1. Change on the size of overhang may cause to dramatically decline or stop of HEN1 methylation acitivity. HEN1 functions regardless of the 5’ end characteristics. In addition to 3’ end nucleotides, 2’ OH of end nucleotide at 3’ end is significant determinant of methyl addition. Methyl is added to this 2’ OH. Efficiency of methylation changes with the length of miRNA, in vitro. Again in vitro, HEN1 has ability to methylate both siRNA and miRNA duplexes. Methylated miRNA enters RISC after processed by Ago for separation of passenger strand from the guide one.

Most of the Ago from Arabidopsis are capable of slicing target RNA. As opposite to animals, plants have miRNAs capable of destructing intended mRNA when perfect or barely perfect matching between mRNA and guide strand is provided. At 3’ end of mRNA, just after translational termination codon, a region called 3’ untranslated region is localized, abbraveated as 3’ UTR. This region contains miRNA response elements (MREs). In mRNA metabolism of plants, interraction between miRNA and MREs determines mRNA will be degradaded or not while in animals, this relation just may cause inhibition of translation.

MiRNA cleavage remains smaller RNA particles which will be degraded by exonucleases, like XRN4. Before exonuclease activity, a few modification like insertion of adeninosine or uridine to the end of target molecule can be required.

Plants utilizes its phloem or/and plasmodesmata to propagate RNAi response to other cells of the plant body. Unlike animal miRNA genes, those ones belongs to plants are intergenic and do not exhibit co- transcription and/or translation. Production of miRNA controlled via feedback mechanism that is mediated by means of miRISC activity on mRNAs of Dcl- 1 and RISC components. It is thought that miRNA evolves fast in plants. This idea is supported with the observation that most of miRNA of Arabidopsis are nonconserved in poplar and rice.

Drosha
In human, DGCR8 and Drosha associates with each other.

Dicer
Insertion of ATP triggers production of siRNA by interraction of ATP with helicase/ATPase domain located on one the Dicer of Drosophila and Dicer of C. elegans. Although mammalian Dicer has helicase/ATPase domain, no influential effect of ATP occurs, in the absence of Dicer in mammalian cells does not inhibits RISC activity.

Two example of dsRNA bindind domained proteins, that associate with human Dicer, are TRBP and PACT. TRBP is an effective component for optimum silencing by siRNAs and endogeneous miRNAs. Additionally, the proten contributesto regulation of some other mechanisms like control of cell growth.

Dicer works under the presence of the Mg2+ which can be replaced with Co2+ or Mn2+. Dicer even can process dsRNA that have blocked ends with RNA tetraloop or DNA- RNA duplexes. It converts these ends to 2nt overhang 3’ ends. After that it continues its normal pathway.

Human Dicer works regardless of ATP. This condition is more or less same for UTP, TTP, GTP etc. and nonhyrolyzable ATP analogs. In chicken- human hybrid cells, lack of Dicer is ended up with the mislocation of RAD1 and BupR1 heterochromatin related proteins.

Efficieny of Dicer- 1 enzyme in Drosophila is dependent to Loqs, a type of protein.

Dicer- 2 from Drosophila melanogaster cannot function without ATP  as other RNA helicases are not able to do, too. Functioning of Girdia intestinalis Dicer void of helicase domain demonstrates dispensability of the domain for working of the enzyme.

An extremely conserved part of RNAase III family proteins is called endonuclease domain. It is found in the form of tightly associated dimer. Because of dimerization, a space form between monomers which dsRNA molecule sits. The space is named “catalytic valley”. The valley carries negative charge on itself which lets magnessium ion to interract. Magnessium ions locate close to ends of this valley.

RNase IIIa domain cuts from 3’ end of one strand of dsRNA while RNase IIIb refers to cut from 5’ end of opposite strand.

In Girdia, distance between PAZ and RIIIDs domains is equivalent to the length of cleavage product. Thus it is thought that length of miRNAs and siRNAs determined by structure of Dicer.

Ago
PIWI domain is the slicer of Ago in RISC. PAZ of Ago contact with 3’ end of siRNA while PIWI domain makes connection both with RNase III domains of Dicer and phosphate headed end of siRNA single strand. PAZ affinity towards 3’ overhangs originated from aromatic residues on it. Different members of Ago family bind to distinct siRNAs.

Intrerraction between Ago and Dicer is achieved by the help of Hsp90 protein. This linkage can be disrupted via block of Hsp90 by means of Hsp inhibitors.

RISC
Size of overhangs is one of the most important factor affecting the RNAi activity. As even change in phoshate head of siRNA may result in severe effects on the mechanism of RNAi, its absence means directly loss of RISC stability.

Available Mg2+ is an obligation for catalytic activity of RISC.

RDRP
RdRP primarily belongs to viruses. It directly contributes to viral gene transcription and replication, in viral systems. In general, RdRP carries 4 main domain, palm ,fingers, thumb and N terminal domain that connects fingers to thumb domain.

It has cleft for sitting of template RNA strand. This template creates base pairing with small part of primer RNA

RdRP, in general, known as a type of error prone polymerase.

Details of RNAi in mammals
Neither sense nor antisense strand of dsRNA alone can trigger RNAi response. Both strands together is needed to do it.

Generally, transcripts of miRNA genes, pri- miRNAs, have at least 1000 nt. After operation of DGRC8/DROSHA on pri- miRNA, pre- miRNAs are gained with 60- 70 nt length, 2nt overhang at both side and 10 nt loop at one side. All of these process happen in the nucleus.

Seed sequence is the region of off-target mRNA that match with 6-7 nt of siRNA sequence. If siRNA on RISC has seed sequence coomplementary to 3’ UTR of mRNA, it behaves as miRNA and prevents transcription of that mRNA, which is not real target RNA molecule. Off target effects could drive cell to death by cellular toxicity. H3K9 and H3K7 of mammalian cells are the targets of methyl addition via siRNA directed mechanism. With this methylation silencing at the level of transcription achieved at related si- RNA targetted promoter sequences.

Vigilin is one of the RNAi protein involved in the relation of RNAi with heterochromatin formation. DNA repair protein DNA- PK (DNA Protein Kinase) behaves like a bridge between vigilin and HP1 (Heterochromatin Protein 1). With its kinase activity, vigilin complex promotes phosphorylation of HP1.

DNA- PK is not only repair protein that vigilin interracts. Another “bridge” protein is ATM which found in DNA Damage Response signaling. Drosophila has ortholog of ATM called dATM that helps HP1 to sit true place on telomeres.

Synthetic shRNAs (small hairpin RNAs), which has 25- 29 nt long and are 2’ overhanged at 3’ ends, are more effective than siRNAs that have the same target sequence. The main problem of the usage of these shRNAs and siRNAs is that they cannot generate permenant or inducible slincing on the cells of mammals. The reason for transient effect is lack steps of amplication process of RNAi involved in another systems. Each cell division and formation of each RISC declines the nnumber of siRNAs whose effect is not amplified. However some solutions are available so as to enable stable gene silencing in mammalian cells.

RNAi machinary is capable of realizing even one nucleotide difference between two mRNA molecule. Because of this high specifity, it can be used for allele- specific purposes means that while it affects mutated allele expression, normal allele expression remains fixed.

As a result of its weight and charge siRNA is not able to pass freely biological barriers. In order to overcome this issue, different methods are developed. 4 main groups of delivery techniques can be counted as:

 * 1)  Lipid based techniques
 * 2)  With high level similarity to lipid based ones, polymer based methods
 * 3)  Naked delivery methods
 * 4)  Peptid based techniques

Succesfull transfection of siRNAs can be reached by electroporation, which causes toxicity, or using microsponge , liposomes or dendrimers.

SiRNAs preferentially created with UU or TT overhangs to be recognized by RNAi mechanism.

2’ – O- (2- methoxyethyl) modification at second base importanly declines off targetting. Utilization of LNAs (Locked Nucleic Acids) as a modification on siRNA is another option to strengthen the RNAi inducer molecule.

Internal modifications are not sufficient at introduction of inducer RNA to central nervous system cells.

Immune system and RNAi
Long dsRNA also triggers sequence independent interferon response (Figure 7 and Figure 8) in addition to possibility of RNAi mechanism induction, in mammalian cells.

Interferons (IFNs) are the cytokines capable of influencing multiple cell types. IFN is dramatically significant at communication among cells throughout innate immune responses beside adaptive ones. It primarily targets bacterial and viral infections alongside neoplastic transformation. INFs catogorized in to type I and type II IFN in terms of the way that they regulate immune components.

Type II IFN contains IFN- γ. IFN- γ sits at the center of immune responses because expression of molecules used in intercellular communication and cellular interractions is under the control of its. At the time of the infection, Dendrite cells and monocytes are alerted by molecules located on the cell wall of the bacteria. Then alerted cells activates T and NK (Natural Killer) cells to produce and release IFN- γ.

MHC (Major Histocompatibility Complex) class I cell surface structures have responsibility to present an order of peptide epitopes to CD8+ T cells. Cells carrying proper epitopes for CD8+ cell recognition interracts with T cell and cause release of IFN- γ.

Released IFN- γ connects its receptor on the target cells and leads dimerization of IFN- γ- R1 and IFN- γ- R2 subunits of the receptor. Further, dimerization causes phosphorylation of JAK1 and JAK2 receoptor associated kinases. Then JAK1 and JAK2 phosphorylates STAT1 monomers to make them proper for dimerization. Dimerized STAT1 goes to inside of nucleus so as to activate transcription of some components found in the immune response.

IFN- γ has a negative feedback mechanism based on SOCS1. SOCS1 which is awaken by IFN- γ, prevents JAK1 and JAK2 activity, thus further steps extending until transcripitional activation fails.

Other machinaries exists in order to regulate IFN- γ signaling pathway. These are mainly based on inhibiting the work of proteins included in IFN- γ signaling and MHC class I path. Block of MHC class I pathway means loss of presentation of abnormal cell to CD8+ T cells, therefore IFN- γ even is not secreted from the T cell.

MiRNAs are also said to be regulatory molecules of innate and adaptive immune system. At many stages of tumor, aberrancies on miRNA metabolism is determined. MiRNAs may contribute or repress the tumor. They achieve this by influencing APM (Antigen Presenting Machinary), which makes protein ready for presentation to immune cells , and regulating IFN- γ signaling proteins.

IFN- γ is able to interfere miRNA metabolism whereas miRNA metabolism can disrupt IFN- γ synthesis, seen in NK cells.

Immune system disorders orginate from overexpression of immune factors. Targetting these factors are able to recover disorders related to immunity.

If the siRNA carries danger motif (5’- GUCCUUCAA – 3’) TLR7 dependent immune response happens via recognition of siRNA by TLR7 receptor on plasmocytoid dendritic cells. Stoppage of this type immune system alertion can be provided by means of chemical modifications or nanopartical coating the inducer RNA. Because TLRs expression is dependent to cell type, immune respose against siRNA is also depends on cell type.

PiRNA
PiRNAs has distinct size and end modification from siRNAs as well as from miRNAs.

PiRNAs’ biogenesis is dicer independent. Activity area of piRNA includes germ- line development, stem cell recovery, transposon blocking and epigenetic regulation. They play role in both posttranscriptional and chromatin level reactions. Though it is not known how piRNAs are produced and amplified, existence of their slicing ability is a known fact.

Long Noncoding RNAs
Long noncoding RNAs (lncRNAs) differ from siRNAs, miRNAs and piRNAs (piwi- interracting RNAs) via their large size (<200 nt).

LncRNAs participates in transcriptional activation, inactivation, silencing, by means of epigenetic modifications, and post- transcriptional gene silencing. Predictions and conclusions from experiments suggest that these RNAs functions by interracting with other RNAs, DNA and proteins, including transcription factors.

Noncanonical RNAi Inducers
While canonical siRNAs are being used, some diverge class of siRNAs discovered which do not alert innate immune system. Noncanonical siRNAs, devoid of any modification, have restrictions in usage arises from their weakness against degradation, probability of undiserable guide strand election and induction of interferon response. These siRNAs are larger than canonical ones while smaller than 30 nt. 27 nt length in these siRNAs gives the best performance at silencing and siRNAs with this length called dicer substrate siRNAs, abbreviated as dsiRNA. DsiRNA are better at generating RISC Loading Complex as compared to canonical ones, which are 21 nt long. One of the crucial issue for in vivo application of RNAi machinary stimulation by dsiRNA is fate of the siRNA in variety of body fluids. Because of this issue RNAs can be modified in a way that does not disrupt Dicer association with modified RNA molecule. 2’ – O- methyl modification is a good and not expensive path to jump over problems relate with stability in body fluids. If dosage of modification is extremely high, mission fails in consequence of loss of Dicer related steps in RNAi system.

Lower concentration of dsiRNA is required for satisfactory silencing, compared to canonical siRNAs. Longer time influence and increased specificity provided via the use of dsiRNAs.

So as to decrease off- targetting, dsiRNA clusters enzymmatically produced via the use of Dicer.

In addition to dsiRNA, other alternatives to canonical siRNA occurs like shRNA, dumbell RNA and fork siRNA (Figure 9).

29 nt stemmed and 4 looped shRNAs are better than 19 nt stemmed ones at silencing of target gene. As loop length is also a significant factor however in high concentrations no critical differences in activity efficiency observed between short and long looped shRNAs. Similar to loop length also side of loop (left or right) is not a determinant of silencing activity at high amounts. Although shRNAs protected at one side via its loop, they even degraded fast in body fluids. This problem overcome by Abe and Abe’s colleagues via invention of new type of siRNA that has two loop at both right and left, called RNA dumbell.

Fork siRNAs are generally preferred to knockout mutated or chimerical genes.

By the help of several strategies long dsRNAs also became a RNAi tool for mammalian cells. They are usually opted for multiple target silencing.

Two grups of long interfering RNA occurs, linear (Figure 10) and branched (Figure 11) longs dsRNA, preferentially with several modification to provide nuclease stability. Some experiments validate these multi siRNAs utilize the same pathway with normal siRNAs.

Linear ones bear a gap on one strand. Devoid of gap, they are non- funtional. Branched ones have more variant shapes.

Although noncanonical RNAi inducers costs much for unit, low amounts give satisfactory results.

Cancer
RNAi theraphies offer to stop drug resistence in cancerous cells via switching on/off the relavant signaling paths. Only RNAi based medication is not sufficient in order to defeat cancer, however, combination with other cancer approaches like radiotheraphy and chemotheraphy, cancer may be knockdown.

Escape Strategies of Viruses
Proteins produced because of viral attact leads formation of negative effects on the metabolism of miRNA. FHV (Flock House Virus) produces B2 that inhibits RNAi via preventing Dicer cleavage. B2 activity result in decline in the number of siRNA.

Some miRNAs are capable of contributing to virus replication while some of them may work for prevention of the replication.

Agricultural utilization
Major reason for agricultural damage is shown to be insect pests. Although Bacillus thuringiensis toxin is highly influensive to combat with some of pests, most of the members of insect pests, especially Hempitera, are not affected by the toxin. Because core RNAi mechinary found in all insect pests, RNAi became a strong option as a pestiside. Generally, one method to knock out agricultural pests is not sufficient because of that Integrated Pest Management (IPM) strategies are preferred.

However not all insect groups contain RNAi mechanism which totally limit RNAi inducer applications in RNAi lacking organisms.

It is essential to conserve beneficial insects, honey bee is one of them, from harmful biological factors, like pathoges and parazites, too.

Conclusion
P bodies also includes RNAi activity alongside cytoplasm.

When use of long dsRNA for silencing target gene unsucessfull in mammalian cells, by innate immune system, in non- mammalians these type of RNAs achieves silencing, because of absence of interferon response. Short siRNA are option to bypass innate immune response. With the shortening of normal siRNA, cleavage point of RISC complex alters, too. However, it is observed that, in some cases, even 21 bp siRNA alongside longer ones can initiate interferon mechanism.

Because miRNA effects comparatively more limited than siRNA influence, usage of synthetic miRNA paralel to natural one also more restricted than synthetic siRNAs.

SiRNA theraphies seem to be more influencive among other antisense theraphies in terms of target specifity, longevity, stability in vivo, efficiency at low amounts and spreading of effect to other cells.

It is thought that usage of RNAi instead of transgenic reformation methods to generate genetically modified foods is safer because RNAi do not use the genes of other organisms. Thus organism genetic code remains orginal.

Learning project: what next?
Read about one of these topics and add what you learn to this page:
 * What if inhibitory double stranded RNA could form from hairpins such as those found in tRNA molecules? (hint)
 * What other medical applications have been proposed for RNA interference? (Hints: search here)
 * Add questions and general discussion to the next sections.

Discussions

 * Read RNAi: a novel strategy for the treatment of prion diseases by Qingzhong Kong and Lentivector-mediated RNAi efficiently suppresses prion protein and prolongs survival of scrapie-infected mice by Alexander Pfeifer et al. Discuss the idea that virus-mediated RNA interference can be used to treat neurological diseases.

Mice as a model system
Are short-lived mice a good model system for study of human neurodegenerative diseases?

title: RNA interference


text:
Fedor Hodža

Fedor Hodža (4 November 1912, in Budapest – 17 September 1968, in New York City) was a Slovak politician and lawyer, the son of Milan Hodža.

He was a graduate of the Faculty of Law of the Charles University in Prague. He worked at the Ministry of Foreign Affairs in Prague (1938 – 1939).

During the World War II (more precisely 1938 – 1945) he lived in France and the UK, where he was Minister of Foreign Affairs and Agriculture of the Czechoslovak government in exile from 1940 to 1945. He participated, along with his father, in the creation of the Slovak National Council in Paris, which was supposed to be an opposition body to the exile policy of Edvard Beneš. After his father had left for the U.S., Fedor established close relationships with Beneš and even became a member of the Czechoslovak pseudo-parliament in exile ("State Council") in London in 1944.

In 1945, he was a member of the Slovak National Council. After World War II, from 1945 to 1948, he was a member of the Constituent National Assembly of Czechoslovakia and Secretary General of the Democratic Party.

In 1948, when the communists took power in Czechoslovakia, he emigrated to the U.S. There, as a member of the Council for a Free Czecho-Slovakia and of the Permanent Council of Slovak Democratic Exiles, he tried to achieve a re-establishment of democracy in Czechoslovakia, ruled by communists at that time. He died on 17 September 1968 in New York City.

source: wikimedia

license: Creative Commons - Attribution Share-Alike - https://creativecommons.org/licenses/by-sa/4.0/

title: Fedor Hodža


text:
A Safe Place

A Safe Place is a 1971 American drama film written and directed by Henry Jaglom and starring Tuesday Weld, Orson Welles, and Jack Nicholson.

Plot
A young woman, named Noah, lives alone in a small apartment in New York City. She is a mentally disturbed flower child, who retreats into her past, yearning for lost innocence. She recalls her childhood, searching for a "safe place." As a child (whose real name was Susan), she met a charismatic magician in Central Park who presented her with magical objects: a levitating silver ball, a star ring, and a Noah's ark.

In the present day, Noah is romantically involved with two different men: Fred, who is practical but dull, and Mitch, who is more dynamic and closer to her ideal fantasy partner. Neither man is able to fulfill her needs totally.

Cast
The cast includes:
 * Tuesday Weld as Susan/Noah
 * Orson Welles as The Magician
 * Jack Nicholson as Mitch
 * Philip Proctor as Fred
 * Gwen Welles as Bari
 * Roger Garrett as Noah's Friend
 * Francesca Hilton as Noah's Friend
 * Richard Finocchio as Noah's Friend

Production
The film was "culled from 50 hours of footage." The work was a product of BBS Productions, a company formed by Bob Rafelson, Bert Schneider, and Steve Blauner, financed by their work on the TV pop group the Monkees. Other BBS films of the era include Easy Rider, Five Easy Pieces, The Last Picture Show, The King of Marvin Gardens, Head, and Drive, He Said. All seven of these films have been restored and released in DVD versions by The Criterion Collection in a set called America Lost and Found: The BBS Story.

Reception
Jaglom's directorial debut was a "critical and box-office disaster" Time magazine called the film "pretentious and confusing", a film that "suggests that the rumors of his expertise were greatly exaggerated, or at least that it does not extend to directing." Vincent Canby described the film as a "superficial case history of a suicide" whose "narrative pretends to be a lot more complex"; Canby noted that the film "reveals the director's apparent adoration of his star [Weld], whom he studies in every possible light and color combination, and in every possible camera setup, often orchestrated with fine, corny songs out of the 1940s and 1950s on the order of Charles Trenet's 'La Mer' and 'Vous Qui Passez Sans Me Voir.'" Variety said the film's "deliberate experimentation puts a heavy burden upon the viewer." Its writer-director "has plunged in over his own depth."

title: A Safe Place


text:
Dutch/Introduction

Layout of the Course
This textbook is intended to be a comprehensive course in the Dutch language for English speakers, but of course people who speak English as a second language are most welcome as well.

Being an Afrikaans speaker is a huge help too, as it is a daughter language of Dutch, though Afrikaans has its differences and words can have different meanings from the same ones in Dutch, and about 95% of Afrikaans vocabulary comes from Dutch. If you are an Afrikaans speaker, this course shall be significantly easier. Just remember the different dialects etc. when speaking Dutch.

Early lessons emphasize conversational subjects and gradually introduce Dutch grammatical concepts and rules. In addition, sound files and illustrations accompany appropriate parts of each lesson.

Requirements for the course

 * Financial: This course does not cost you a dime.

 * Audio: Listening and speaking yourself are an important part of language acquisition. Therefore there are many sound files embedded in the course, so do make sure your computer or other device can play them. This includes using an appropriate browser. The author is using Firefox. Chrome might work too, but Internet Explorer does not always show the sound buttons.

 * Flashcards: We have added links to Quizlet data sets in parallel with each lesson through lesson10. Thus, all 24 lessons of the Beginner Level have Quizlet links. Registration for Quizlet is recommended (It is for free).

 * Keyboard: We also recommend that you install an international keyboard to deal more easily with the -rather few- diacritics that Dutch uses. For Microsoft operating systems this will tell you how to proceed.

Structure of the course
The course is divided in three levels: beginner, intermediate and advanced. Each level consists of two cycles of 4 lessons each. At the beginner and intermediate level each lesson is accompanied by two parallel lessons: a practice lesson and a cultural lesson. That means that the beginner level comprises 2*4*3= 24 lessons in total.

At the advanced level there are no parallel lessons. Starting from scratch, expect to arrive there in a year, depending on how intensively you study. By that time you should be able to start picking your own things to read, newspapers, Wikipedia articles, what have you.

Parallel lessons
The main lessons aim at introducing grammatical topics by means of conversations, interspersed with some exercises. Of course that is not sufficient to actually start speaking the language. Therefore, each lesson is accompanied by:
 * A parallel practice lesson 1⇒1A that elaborates the material further in conversations, reading material, fill in the blank exercises, small quizzes, pronunciation drills etc., to practice the material in the main lesson and build up vocabulary.
 * A parallel example lesson 1⇒Vb. 1 that uses pieces of literature, songs, nursery rhymes at times linked to YouTube videos to expose the reader to some Dutch cultural phenomena. In the earlier lessons the material is originally mostly intended for children who are acquiring the language in their playful way. In later lessons the material becomes a bit more adult and involves poetry and other forms of literature and some cultural and historical background information.

In addition there are pages intended to help build up vocabulary, some of which are interwoven in the practice lessons. Other ones are stand alone.

Possible strategies
Which way the reader wishes to use the book may vary. The recommended strategy for a beginner with no experience is:

Lesson 1 > Lesson 1A -> Vb. 1 -> Lesson 2 > Lesson 2A > Vb. 2 >Lesson 3 > Lesson 3A >, etc.

People who have experience with other languages, grammars etc. might want to follow the order

Lesson 1 > 2 > 3 > 4 > and on to the end of the basic text

Others may want to start tackling the language in context of a situation and worry about grammar later might want to start with Lesson 1A or Example 1 and check back for the grammar in Lesson 1 later. But readers are encouraged to revisit pages they have worked on again and again, so the order may be more complicated than this.

For people who have been learning Dutch in other ways and want to revisit a certain topic there is an Index of grammatical and syntactic topics covered.

Layout within main Lessons
The following topics can typically be found in one of the main lessons:

 * 1) One or more conversations (Gesprek) or stories (Verhaal) in a colored box, usually with a sound button
 * The translation of these textual pieces is given in a drop down box. Words can also be underlined, and the reader can hover the mouse over.
 * A vocabulary box on the right hand side that can be opened for easy access to pronunciation, translation and links to WikiWoordenboek
 * 1) One or more grammar (Grammatica) lessons covering elements of Dutch grammar, with examples often drawn from the conversation, story, or study materials.
 * 2) Sections (''Uw beurt - Your turn) with exercises pertaining to the material of various kinds:
 * Fill in the blank
 * Pronunciation drills
 * Translation assignments
 * Modifying sentences
 * 1) Illustrations of concepts related to the story usually with a sound button
 * 2) A list of words (Woordenschat) and phrases introduced in the lesson, usually in the conversation, story, or study presentations. including sound files
 * 3) A link to Quizlet with the vocabulary taught (under construction)
 * 4) A synopsis of what the reader should have gained, including a cumulative vocabulary count (under construction)

Layout in the practice lessons
There are often additional texts, many exercises of various sorts and short quizzes to practice what was taught in the main lesson or to repeat what was taught in the lessons before. To further expand vocabulary students may be asked to go to one of the vocabulary pages to study words related to a certain topic. Quizlet links provide another way to brush up on vocabulary

Layout in the example lessons
In these lessons we will have a look at rhymes, poems, songs etc. Often the student is referred to a YouTube video to watch in order to practice oral understanding and expand vocabulary in the context of Dutch culture. Quizlet links provide another way to brush up on vocabulary.

Quizzes
Short quizzes are integrated into the practice lessons, but there are also longer quizzes at the end of the two cycles of the Beginner Level.

Rate of acquisition of vocabulary
Each lesson -its two parallel lessons included- contains on average about 120 new terms. At the end of the Beginner Level the student should know a little over one thousand terms.

The Student and the Lesson
The text is designed to constitute a comprehensive course of study in the Dutch language. Each lesson should be read thoroughly and mastered before moving on. Substantial text in Dutch is included and the student should read all of it, not once, but multiple times. In most cases sound files are given as an arrow button; they should be listened to multiple times as well, both while reading the text simultaneously, and -once the content is understood- with eyes closed. Complete translations into English are included only in selected places or they are hidden in a drop down box. Most of the text should first be translated by the student using his or her acquired vocabulary and the vocabulary presented at the bottom of each lesson and/or in collapsed boxes in the right hand margin of the text. Hints about the meaning of new, underlined words can also be obtained by the.

As the Dutch is read (out loud is better), the student must succeed in gaining an understanding of the meaning of each sentence, and the role each word plays in establishing that meaning. To the beginner, there will seem to be many words in a Dutch sentence that are out of place or even redundant or unnecessary. These add subtleties to the language that will make sense eventually. But it is important to experience these subtleties from the very beginning.

There are exercises interspersed throughout the main lesson with additional ones in the practice lessons. They are of various nature. Translation exercises, pronunciation drills, fill-in-the-blanks, adapt a word form, swap to items etc.

The Dutch Language
Dutch (Nederlands) is a member of the western group of the Germanic languages. It is spoken primarily in the Netherlands, and in a major part of both Belgium and Surinam. It has about 23 million mother tongue speakers and another 5 million second language speakers. Continue reading about the Dutch language and its history at Wikipedia.

Dialects
As a standard language Dutch is a relatively young phenomenon. The standard is based on a variety of dialects that are much older and show considerable differences not only in pronunciation but even in grammar and syntax. This holds for many languages, including for English as spoken in the UK. By urbanization, suburbanization and the influence of the mass media the standard language has been gaining ground at the cost of the dialects for over a century, so that it is now the mother tongue of most speakers. Others are typically perfectly bilingual in their regional tongue and the standard language. But in the way that the standard is spoken there are many regional and social differences in pronunciation or even in syntax and grammar. In Bruges (Flanders), Rotterdam (Netherlands) or Paramaribo (Suriname) Dutch will sound as different as English does in Edinburgh, London or Indianapolis. This course aims at teaching Dutch that would be acceptable to most if not all speakers but will point out a number of important differences that non-native speakers are likely to encounter in their interaction with native speakers.

A major division in the dialects is formed by de grote rivieren the great rivers that run through the Netherlands from east to west on their way to the North Sea as shown in the map. The course is mostly based on Northern standard Dutch, because that is most readily accepted under all speakers, but it will point out some important differences at times and also give examples of other varieties.

A dynamic language
Dutch has undergone far more sweeping changes in grammar and syntax in the last century or two than either English or German. It has lost most of its case endings and much of one of the three original genders (feminine). This has led to some interesting shifts in its grammar and syntax. Some of these developments are still taking place today. This means that Dutch grammar is less set in stone than the reader may be familiar with from other grammars. Occasionally we will have to discuss the evolution rather than the creature to explain modern Dutch usage.

Dutch and English
If you are an English speaker unfamiliar with Dutch, you may be surprised to learn that English and Dutch are closely related languages and share many words that are very similar. This is particularly true for everyday words in English that are Anglo-Saxon (i.e. Germanic) in origin. After 1066 English has absorbed a lot of (Norman) French. Dutch also has been exposed to contact with first vulgar Latin and then French, but the French influence has been less pervasive.

Consider the following list of English words followed by their Dutch counterparts: Many words of French origin have entered both languages and are quite recognizable:

 * communication ~ communicatie
 * proclaim ~ proclameren

But in many cases Dutch retains a Germanic word, sometimes aside the Latin one:

 * proclaim ~ uitroepen

English spelling has conserved many now silent consonants, e.g. gh in light. This may have been an obstacle when learning to write English but when learning Dutch the investment pays off. Dutch has licht and the ch is very much still pronounced as a guttural fricative /x/ like in German Bach or Scottish Loch.

Some words are even completely the same. (true friends)

Of course, even words whose spelling is no different in English and Dutch may be pronounced quite differently or mean something different (false friends):

 * door ~ deur
 * through, by ~ door

 * worst ~ ergst, slechtst
 * sausage ~ worst

Nevertheless, when reading Dutch you will see the kinship between the languages, even in many short words, common or not. For example, compare:

 * This week, my father is on the wharf with my brother and his daughter.
 * Deze week is mijn vader op de werf met mijn broer en zijn dochter.

These sentences consist almost entirely of cognates: words that evolved from the same source. Dutch is indeed one of easiest languages to learn for an Anglophone. Notice however the position of the verb is in these two phrases. In Dutch it stands in front of the father. This is because Dutch has retained something that English has lost: the rather complicated word order (syntax) of the West-Germanic languages. Many English speakers who learn Dutch find that one of the most difficult aspects to learn to do correctly, but it hardly ever leads to miscommunication. In the course it is introduced bit by bit. The full picture is only described in one of the last lessons (21).

Dutch and German
Both Dutch and German are West-Germanic languages and this means that there are many resemblances. However, Dutch is easier to learn for a speaker of English for a number of reasons. First of all, (High-) German underwent a major shift of almost all its consonants in the early Middle Ages. In term of its consonants Dutch has been pretty conservative. Compare: This makes a major part of Dutch vocabulary easier to memorize. Secondly, German retained its system of case endings in contrast to Dutch and English. It is not easy to master that system if your mother-tongue does not have it. Compare:

Knowledge of German can certainly help in learning Dutch, but it can also be a source of confusion. A good example is the letter combination sch. In German it denotes the same consonant as sh in English (in IPA: [ʃ]), in Dutch this sound is relatively rare. It only occurs in loans from languages like Frisian, English, French etc.

In Dutch 'sch' usually denotes [sx]: an [s] followed by a velar spirant [x], like in schip. In the ending -isch the 'ch' is mute and it is pronounced as [-is] as in English 'fleece'.

A topic where knowledge of German is a great help is syntax (word order), but on the other hand there are differences in how the verb tenses are used. In German the imperfect past tense, like du gabst, sprachst, rettettest is on its way out. In Dutch forms like jij gaf, sprak, redde are alive and kicking and in every day use.

Dutch and Scandinavian languages
Although Danish, Swedish and Norwegian are North-Germanic languages, which means that the relationship is a bit more distant, speakers of these languages typically do not find Dutch very difficult, because many changes in the language such as the loss of the case endings and of feminine gender are quite similar. Some Scandinavians manage to speak Dutch so well without any discernible accent that people don't realize that they are not Dutch. Swedes may have to pay attention to their intonation, because that is rather different in Swedish.

Dutch and French / Italian and other romance languages
For speakers of the Romance languages Dutch is by no means an easy language to learn, although if you already speak English some of the problems may already have been overcome.

Dutch is a stress language like English and German. That is: every word has one syllable that is high in pitch, a bit louder, and usually a bit longer than all the other ones. In French those three qualities are not coupled and spread more evenly over the syllables of the word. In Dutch, stressed syllables have either a full vowel (one of twelve) or a diphthong; the unstressed syllables all tend to have a schwa. Intonation is therefore difficult for Romance speakers because the contrasts between the syllables is much smaller in those languages.

Another problem is formed by the separable verbs. There often is no direct equivalent in French for the fine nuances imparted by the separable prefix in Dutch. Often an entirely different verb needs to be substituted, or there is none available.

French speakers usually have little problem with the vowels, but they do tend to speak much more in the front of the mouth than Dutch speakers do. The placement of the sounds is different. For other Romance languages a vowel like u [y] or eu [ø] the ui [ʌy] diphthong might be problematic, as well as the guttural spirants g and ch.

Dutch and Russian / other Slavic languages
A main problem for Russian speakers lies in the vowel system of Dutch. The vowels a, e, i, o, u all occur in two different qualities. The differences are difficult to hear and reproduce for Russian speakers.

Another problem is the voiced consonants like b, v, d, z. In Dutch they all get devoiced at the end of the word, but that hold for Russian too. However, they often also get devoiced when they follow a voiceless consonant, even in assimilation from the previous word. E.g.
 * de vele autos - [v]
 * het vee  - [f]

A grammatical problem is that Dutch is a tense language and Russian an aspect language. Dutch has perfect tenses, Russian perfective aspects. They cover the same territory, but one lengthwise the other in the other direction.

Dutch and Mandarin
Of course the differences are very large and numerous, but one difficulty deserves mention, that of epenthesis. The syllables of Mandarin are either CV or CVn, i.e. they start with a single consonant and end in a vowel or a nasal. In Dutch they can end in other consonants and in fact multiple ones, like herfst is CVCCCC with no less than four consonants at the end. Mandarin speakers need to suppress the tendency to add epenthetic vowels, making it something like herefesete. Epenthetic (inserted) vowels are sometimes used by Dutch speakers too, e.g. werken may well be pronounced werreke, but this is considered dialectal and non-standard and is frowned upon by most speakers.

Vocabulary and Grammar
In learning to read or speak any new language, two important aspects to be mastered are vocabulary and grammar (others are pronunciation and syntax, but they usually do not stop you from being understood). Acquiring vocabulary is a "simple" matter of memorization.

Learning by ear
Children do it all the time, but they are at an advantage: they memorize far easier than grown-ups. Age is a definite disadvantage in language learning. The child's learning process can be "reactivated" to some extent by immersion in a second language: a method of learning a new language by moving to a place where that language is spoken and having to get around and live without use of one's native tongue.

If you do not have the opportunity of residing in a Dutch speaking area an alternative is to listen to recordings and we are in process of adding bits and pieces as .ogg files so that you can learn by ear. Use them as much as you can. More than once.

These files take different forms
 * 1) Single words. They are useful when you are trying to memorize vocabulary
 * 2) Spoken text of the chapter. They should be used to study the conversations
 * 3) Drills. Here you need to repeat words or utterances in the pauses.
 * 4) Translation drills. Here you are told to say something in Dutch yourself.

In the Example lessons there are also links to YouTube videos where the text of a poem is recited or a song is sung.

Of course there is also a drawback to the by-ear method: You do not get much immersion into reading Dutch. You as an internet user, will most likely want to be literate in Dutch.

As with all languages: the written Dutch language and the spoken Dutch language are by no means identical. At times we will explain the distinction if necessary.

Learning by eye
This is why this course also tries to train your eyes, but this will not work without effort from your side. This is why we often say: Your turn! (Uw beurt!)

So what do you need to do? There are a variety of things. We are tackling the problem with a multi-pronged approach.

Be sure to "learn"—commit to memory—all of the vocabulary words in each lesson as they are presented. Early lessons have simple sentences because it is assumed that the student's vocabulary is limited.

To help you accumulate vocabulary there are a number of additional pages see: Dutch/Vocabulary. They are mostly both visual and audio in nature and there are exercises to go with them (still being created).

Throughout the text, more complex discourses (e.g. as photo captions) are included to introduce the student to regular Dutch in use. It may be helpful to translate these using a Dutch-English dictionary (Wiktionary is usually on a click or two away). Other sources of Dutch, such as newspapers, magazines, web sites, etc. can also be useful in building vocabulary and developing a sense of how Dutch words are put together. The Dutch Wikipedia provides an ever-expanding source of Dutch language articles that can be used for this purpose. Further, a Dutch version of the English Wikibooks project—a library of textbooks in Dutch — is available at Dutch Language Textbooks and there is a growing Dutch version of wiktionary (WikiWoordenboek) to which a number of words in the text have been linked for direct reference. WikiWoordenboek usually has an example phrase to go with every dictionary entry to show the word in context. This too is a helpful tool for expanding your vocabulary, as context helps memorization.

Learning grammar and syntax
This is where as a grown up you are at an advantage, because you may already know how grammar works from your mother tongue or other languages you are proficient in to some extent.

Dutch grammar is sufficiently similar to English grammar that "reading" Dutch is possible with minimal vocabulary. The student should generally recognize the parts of a sentence. With a good dictionary, a sentence can usually be translated correctly. Of course there are some notable exceptions and false friends, e.g. in the way that the passive voice is formed:

 * hij wordt gezien - he is seen
 * hij is gezien - he has been seen.

To speak and write Dutch properly you do need to learn its grammar and syntax. Particularly the latter (word order) is rather different. We will gradually introduce it. Do not be daunted by it. Learning a language goes bit by bit, word for word, structure by structure. Just keep at it and look at what you have gained not at what you don't understand. Children don't always understand everything either, but they are not ashamed or humiliated by that.

Pronunciation
A guide to pronunciation of Dutch is provided as Appendix 1. You should become familiar with this page early on, and refer to it often. Nothing can replace learning a language from a native speaker, but the text is liberally sprinkled with audio files providing the student with valuable input from hearing spoken Dutch. Analyze the spoken words carefully. Descriptions of the pronunciation as in Appendix 1 can only closely, not exactly, convey how Dutch words should be pronounced. This is why there are quite a few buttons on that page that allow you hear what is meant. Do compare!

Of course there are variations in pronunciation. Dutch spoken in the country side of Brabant, in the cities of Amsterdam, Antwerpen or Paramaribo sound pretty different and the same thing can be said for the board room and the back alley. We do present pronunciations from different people and places in the sound files and even more so in the YouTube video links.

Contact
If you have questions or want contact try the dutchgrammar forum. I often hang out there and correct people's stuff or do some exercises

title: Dutch/Introduction


text:
بلدية

Etymology 1
, from the root.

Noun

 * 1) township, rural community
 * 2) district, ward (of a city)
 * 3) municipality
 * 4) municipal council
 * 5) city hall

title: بلدية


text:
Wikipedia:Mediation Cabal/Cases/2006-08-10 Leó Szilárd

Mediation Case: 2006-08-10 Leó Szilárd
Please observe Etiquette and Talk Page Etiquette in disputes. If you submit complaints or insults your edits are likely to be removed by the mediator, any other refactoring of the mediation case by anybody but the mediator is likely to be reverted. If you are not satisfied with the mediation procedure please submit your complaints to Wikipedia talk:Mediation Cabal.

Request Information

 * Request made by: Hi There 01:42, 11 August 2006 (UTC)

Hi There
 * Where is the issue taking place?

http://en.wikipedia.org/wiki/Le%C3%B3_Szil%C3%A1rd
 * Who's involved?

Hi There, Will314159, SafeLibraries
 * What's going on?

Dispute concerning the appropriateness of inclusion of certain highly contentious historical issues which are not necessary for the article to include. Edits and reversions, with my opponent reverting but refusing to discuss the matter on the talk page, and labeling my changes as vandalism on the History page.

1) I removed section about the decision to use the bombs and the effects of their use from the article. The removed section posited certain very debatable theses and relied on ONE book by ONE author which the article thereby implied was authoritative. I removed this section for reasons which I specified on the Talk Page as follows: "I have completely removed the sections concerning the effects of the atomic bombings and the reasons for the decision to use the bombs. It is a highly contentious issue and is being debated at length on the WP page about the bombing (Atomic bombings of Hiroshima and Nagasaki). There is no point to having a stunted rehash of that article here too, and any interested reader would do MUCH better to refer to the article - which, luckily, is already cited in the text itself." I replaced the text I removed with the advice to go read the WP article on that subject specifically and included a hyperlink. Another person interested in the Szilard article, User SafeLibraries, agrees with this edit.

2) User Will314159 reversed my edit, not discussing it on the Talk page, merely labelling his edit on the History page "atomic bomb too close to subject's hear to minimize"

3) Considering this to be insufficient reason to reverse my edit, and having had answers to none of my objections on the Talk Page, I restored the edit that Will314159 had undone.

4) User Will314159 then reversed my edit, not discussing it on the Talk page, merely labelling his edit on the History page "Quit it Hil There you are trivializing man's lilfe" (Original spelling preserved)

5) I restored the my edit (which User Will314159 had removed) and I removed the section that he had reinserted. I put the following comment on the Talk page: "I am removing the section about the effects of and decision to drop the atomic bombs again. My previous remarks and reasons for removing the section are still valid; if you think otherwise please explain why. If you think that James Carroll's opinion is final and definitive, please support such a contention, which you could do, by, let's say, going to http://en.wikipedia.org/wiki/Atomic_bombings_of_Hiroshima_and_Nagasaki#Debate_over_the_decision_to_drop_the_bombs and citing Carroll there - and if all the participants in the discussions THERE accept Carroll's opinion as final and definitive and if all further debates therewith cease in deference to his opinion, then perhaps it will accepted here too; but if not, not."

6) Once again User Will314159 reversed my edit, and in lieu of discussing it on the Talk Page, merely contented himself with labelling my changes as "Vandalism" on the History page.

 * ''What would you like to change about that?

1) I would like the opinion of an uninvolved third party as to the appropriateness or otherwise of my editorial decision to remove the passage the originally removed, as having the implication that the book and author cited by User Will314159 have an authoritative opinion in the matter of the history of the atomic bomb. I also want the passage deleted as being (as I said) far too controversial and contentious to be discussed in anything other than a dedicated article, and certainly not to be discussed in a short biographical article.

2) If my edits are appropriate, I would like User Will314159 enjoined from making any more changes in the article, especially because (but not only because) he feels no need to discuss anything. (And frankly, this is a 0-sum game; I can only accept a link in the article to the complete WP article on the bombs, whereas he seems to think that the topic is simple enough to sum in a short paragraph and that he has found an author whose opinion is final, decisive, and beyond disputation.)

 * Would you prefer we work discreetly? If so, how can we reach you?

No, do it in public, so that we may all be enlightened. So a third opinion placed on the original Talk page of the disputed article would be good.

Mediator response
Working on it. This case seems to be lacking any meaningful communication on the part of one of the parties involved. Torinir  ( Ding my phone  My support calls   E-Support Options  ) 20:58, 11 August 2006 (UTC)

Interest in the case died fast and hard. Overlying issue seems to have been resolved outside of MC. Closing. Torinir ( Ding my phone  My support calls   E-Support Options  ) 01:18, 8 November 2006 (UTC)

Compromise offers
This section is for listing and discussing compromise offers.


text:
Cookbook talk:Baking

This module uses text adapted from the Wikipedia article Baking. The authors at the time the text was copies are inlcuded in the page history below:

* (cur) (last). . M 06:52, 6 Dec 2003. . Steeev (The person that does the baking is called a baker.) * (cur) (last). . 13:22, 21 Jul 2003. . <IP_ADDRESS>   * (cur) (last). . M 13:46, 10 Jul 2003. . Spud007 * (cur) (last). . 12:12, 7 Jul 2003. . <IP_ADDRESS>   * (cur) (last). . M 11:48, 23 Mar 2003. . Bluelion (link) * (cur) (last). . 00:09, 16 Oct 2002. . Merphant (copyedit) * (cur) (last). . 16:44, 30 Sep 2002. . <IP_ADDRESS>   * (cur) (last). . 16:44, 30 Sep 2002. . <IP_ADDRESS>   * (cur) (last). . 16:43, 30 Sep 2002. . <IP_ADDRESS>   * (cur) (last). . M 14:32, 1 Sep 2002. . Ben-Zin (de:) * (cur) (last). . M 10:40, 8 Jul 2002. . Karen Johnson (link dessert, cake) * (cur) (last). . M 08:11, 20 May 2002. . Ap

title: Cookbook talk:Baking


text:
Thom Pain (based on nothing)

Thom Pain (based on nothing) is a 2004 one man show written by Will Eno. It is a rambling monologue in which the protagonist, who has suffered a lot in his life, tells the story of a bee sting, a boy with a dog that died, and his experience with a woman.

Production history
The play started as a reading at the Soho Theatre and was then performed at the Edinburgh Festival Fringe in 2004. The play next opened in London at the Soho Theatre, running from 3 to 24 September 2004. It received the First Fringe Award.

Thom Pain (based on nothing) premiered in the United States Off-Broadway at the DR2 Theatre on February 1, 2005 and closed on September 4, 2005. The original production starred James Urbaniak, and was directed by Hal Brooks. Urbaniak was nominated for the Drama Desk Award for Outstanding Solo Performance for his portrayal of Thom Pain.

The play was revived at the Off-Broadway Signature Theatre Company on October 23, 2018 (previews), starring Michael C. Hall and directed by Oliver Butler.

The play has been translated into over a dozen languages, and since 2005 has been produced regularly around the U.S. and the world. Some notable American productions have been presented at Seattle Rep, the Dallas Theater Center, Theatre Wit, Hyde Park Theatre, Actor's Express, among many more. Worldwide the play has appeared in Finland, France, Israel, Holland, Norway, Portugal, China, Mexico, Germany, Greece and elsewhere.

The play was adapted into a feature film starring Rainn Wilson in 2016. The film adaptation was directed by Will Eno and Oliver Butler, edited by Tymon Brown, and produced by Geffen Playhouse.

The play was a finalist for the 2005 Pulitzer Prize for Drama, which was won by Doubt, A Parable by John Patrick Shanley.

Critical reception
The play received enthusiastic reviews. David Gritten of the Telegraph  wrote, "It's hard to imagine more dazzling writing on any stage in Edinburgh this year."

The Theatre Journal wrote: "Why did this production receive some of the strongest reviews of the season and a twice-extended run in the heart of New York City? The production aesthetics seem to work against popularity."

Charles Isherwood, in his review for The New York Times wrote: "It's one of those treasured nights in the theater - treasured nights anywhere, for that matter - that can leave you both breathless with exhilaration and, depending on your sensitivity to meditations on the bleak and beautiful mysteries of human experience, in a puddle of tears. Also in stitches, here and there. Speechless, in any case."

The Theatremania.com critic wrote: "At its conclusion, audience members may not know exactly what they just saw or what it was supposed to mean. Is Thom Pain a meditation on disappointment? An exercise in futility? Perhaps both. One thing is certain, however. Even if the piece is based on nothing, as its subtitle proclaims, its unconventional style makes it far more interesting than many other plays that are currently on the boards."

Ten years after the original production, Lyn Gardner of the Guardian wrote, "it's still one of the best monologues I've ever seen".

title: Thom Pain (based on nothing)


text:
Wikipedia:Peer review/List of municipalities of Portugal/archive1

List of municipalities of Portugal
I started creating this list inspired on the already featured List of municipalities of Lithuania. It has a similar structure and is giving me lots and lots and lots of work because Portugal has 308 municipalities and I decided to put 5 variables, which gives 1,540 different values, plus the ranks. Along with that, in 14 districts (out of 20), I only added the info after including the Portuguese municipalities infobox in all of its municipalities. And also the maps, which I made by hand. I really want to make it featured, so, I need your help. Make your review, and if you want, answer to my questions:
 * Do you think the article needs ranks? (I've got the data in an Excel file, ready to be included)
 * Do you think it has good references?
 * Do you think it needs better maps?
 * Do you think it is well organized?
 * Do you think it can reach FL status?
 * Do you like the ruling party color system?
 * (The district of Vila Real lacks the data, I'll put it soon)

Thanks! Afonso Silva 00:48, 20 March 2006 (UTC)
 * I think the maps are sufficient and it is well-organized, but why are density and km2 linked each time? Thanks, AndyZ 02:37, 20 March 2006 (UTC)
 * It was copy paste, I did not spot that, I can remove that. Thanks! Afonso Silva 10:58, 20 March 2006 (UTC)


text:
Talk:Fresnel diffraction

 * Please put new material at the bottom of this page. Otherwise, since frequent readers go to the bottom to see what is new your contributions will often be missed.

fresnel number and approximation
On the page about fresnel's number, it says that this equation is only accurate when its greater than or equal to 1. Is this correct? On the page about diffraction, it says that it describes the more general case of diffraction when compared to far-field diffraction. These are contradictory, and this page only says its used when the wave is "a small number of wavelengths away from the diffracting object". Is it an approximation or genralization? Fresheneesz 06:02, 20 March 2006 (UTC)

In fact, the Fresnel approximation should be good whenever $$\frac{N_F \theta_{\rm{max}}^2}{4}\ll 1$$. $$N_F$$ can be smaller than 1, but doesn't have to like in the far-field approximation. --Ahmes 14:37, 3 July 2006 (UTC)

Alternative Forms: Fourier Transform
I do not agree with the following section, which states that H is the fouriertransform of h

 * $$= H(p,q) \cdot G(p,q) |_{p = {x \over \lambda z}, q = { y \over \lambda z } }$$

where
 * $$H(p,q) = \mathcal{F} \left\{ h(x,y) \right\} $$

This can not be true, since H is equal to h. —Preceding unsigned comment added by <IP_ADDRESS> (talk) 10:11, 2 February 2009 (UTC)

Undefined Values, unclear Fourier Transform
In this text

 * $$G(p,q) = \mathcal{F} \left\{ g(x,y) \right\} \equiv \iint_{-\infty}^{\infty} g(x,y) e^{-i 2 \pi (p x + q y)} dx dy $$

where p and q are spatial frequencies (in units of lines/meter). The Fresnel integral can be expressed as:

 * $$E(x,y,z) = \frac{e^{i k z}}{i \lambda z} e^{i \frac{\pi}{\lambda z}(x^2 + y^2)} \mathcal{F}

\left. \left\{ E(x',y',0) e^{i \frac{\pi}{\lambda z} (x'^2 + y'^2)} \right\} \right|_{p = \frac{x}{\lambda z}; q = \frac{y}{\lambda z}} $$

g(x,y) is used without any definition. Also it is not clear why the FT is done on
 * $$E(x',y',0) e^{i \frac{\pi}{\lambda z} (x'^2 + y'^2)}$$ —Preceding unsigned comment added by <IP_ADDRESS> (talk) 10:28, 2 February 2009 (UTC)

Photos
The picture given as an example for fresnel diffraction should be changed, because it gives you a wrong idea about the phenomenon. <IP_ADDRESS> (talk) 13:22, 29 May 2009 (UTC)
 * What do you mean? It is consistent with a whole page full of fresnel diffraction photos given by Francis Weston Sears in his ''Optics," which is a standard text. It is not as nice a photo. If anyone has access to a better photo that shows a center black spot, or a black spot and black rings, let them supply the better picture. If you don't believe Sears (even though Sears and Zemansky is still a standard physics text), look at the simulation software that is linked to the article.P0M (talk) 07:48, 30 May 2009 (UTC)
 * OK, i took some shots and i'll try to upload them now. <IP_ADDRESS> (talk) 20:49, 14 July 2009 (UTC)
 * I had no tripod, so the picture is a bit unsharp, i will do better ones in 3 weeks. —Preceding unsigned comment added by <IP_ADDRESS> (talk) 20:53, 14 July 2009 (UTC)

Why is the picture i uploaded not visible any more? Always a beginner (talk) —Preceding undated comment added 14:21, 7 October 2009 (UTC).
 * What often happens is that somebody uploads a photo but does not give information about it. Specifically you need to state that it is your own work (by putting your user name in as "author," for instance), and you need to choose a version of the permission to reproduce from the GFDL drop-down menu. One of them is marked as recommended, so I usually choose that one as default. So all you really are doing is to say that you made the photo and that you give it away. Also it is best to give a description, i.e., say that it is a pattern produced by fresnel diffraction, the hole is .3 mm in diameter (or whatever it actually was), etc.  In other words you supply all the technical stuff that is relevant to the physics of the phenomenon in question.
 * If I fail to fill in required information I generally get a warning on my user page telling me the photo will be deleted if I do not fix things soon. Since you are not a registered user, you probably never saw any notice, and so the administrators saw a photo that was possibly a copyrighted image copied from someplace without permission, and they deleted it. P0M (talk) 22:59, 7 October 2009 (UTC)

Variable Definition
Many variables appear on this page, but not all of them seem to be defined. Some of the variables I have noticed include "a" and "L".

1/R^2 Dependency of field instead of 1/R
In the general form of the integral for the field at a given point, the expression involves a 1/R^2 dependency of the field. It should actually be a 1/R dependency on distance. see here for derivation: http://www.cv.nrao.edu/course/astr534/PDFnewfiles/LarmorRad.pdf I edited it. Please tell me if I'm doing things inappropriately as I am new to editing wikipedia. Thanks! Edit: just realized that the page is dealing with the near field diffraction pattern; different sources call the Fresnel pattern differently; some call it the far field and others call it the near field pattern; that's where the 1/r^2 difference came from. edited back. — Preceding unsigned comment added by Buhbuhbuhbowser (talk • contribs) 23:13, 2 May 2012 (UTC)

I'm with you on thinking this should be 1/r...? Why wouldn't the 1/r equation apply in the near field too? See for example http://www.iue.tuwien.ac.at/phd/kirchauer/node50.html eqn 4.4 Pwrichards (talk) 21:40, 11 May 2016 (UTC)

It should be 1/r, compare it just to the last equation in the paragraph, there it it also 1/r (or 1/z). See also equation 3.1 in Schnars, U. & Jüptner, W. P. O. Digital recording and numerical reconstruction of holograms. Meas. Sci. Technol. 13, R85 (2002). Low87 (talk) 10:59, 1 July 2016 (UTC)

title: Talk:Fresnel diffraction


text:
Puzzles/Easy Sequence 9/Solution

These numbers are sorted by reverse alphabet:

two, three, six, seven, one, nine, four, five, eight

2 3 6 7 1 9 4 5 8

title: Puzzles/Easy Sequence 9/Solution


text:
Brew (surname)

Brew is a surname. Notable people with the surname include:

 * Aled Brew (born 1986), Welsh rugby union player
 * Bill Brew (1918–1941), Australian rugby league footballer
 * Charlotte Brew, British equestrian
 * Chartres Brew (1815–1870), Canadian judge
 * Chloe Brew (born 1995), British rower
 * Derrick Brew (born 1977), American sprinter
 * Dorian Brew (born 1974), American football player
 * Francis Brew (cricketer) (1903–1974), Australian cricketer
 * Frank Brew (1927–2020), Australian rules footballer
 * George G. Brew (1868–1937), member of the Wisconsin State Assembly
 * Helen Brew (1922–2013), New Zealand actress, filmmaker and educator
 * John Brew (born 1938), Australian businessman
 * Josephine Macalister Brew (1904–1957), British educator
 * Kate Brew Vaughn (1873–1933), American author and teacher
 * Kwesi Brew (1928–2007), Ghanaian poet and diplomat
 * Nathan Brew (born 1982), Welsh rugby union player
 * Neil Brew (born 1979), New Zealand rugby union player
 * Paul Brew (born 1965), British swimmer
 * Rama Brew, Ghanaian actress
 * Ray Brew (1903–1979), Australian rules footballer and coach
 * Robin Brew (born 1962), British swimmer
 * Thomkins Brew, Irish resident magistrate

title: Brew (surname)


text:
Misato (surname)

Misato (written: 美里 or 美郷) is a Japanese surname. Notable people with the surname include:

 * Aki Misato (美郷 あき), Japanese singer
 * Misato Anman (美里 親方 安満), Ryukyuan politician and bureaucrat
 * Misato Chōtei (美里 王子 朝禎), Ryukyuan prince

title: Misato (surname)


text:
competitivity

Noun

 * 1)  competitiveness
 * 2) * 1962, "Britain's Export Trade with Canada. Canadian Studies in Economics: No. 12" (Review), Angus Maddison, The Economic Journal, Vol. 72, No. 286. (Jun., 1962), pp. 395, URL: JSTOR
 * It would throw further light on the inter-relation of competitivity and growth if we had a similar structural analysis of British performance in a more slowly performing market.
 * 1) * 2000, Sylvain Gandon and Yannis Michalakis, "Evolution of Parasite Virulence against Qualitative or Quantitative Host Resistance," Proceedings: Biological Sciences, page 986, Vol. 267, No. 1447. (May 22, 2000), pp. 985-990, URL: JSTOR.
 * Parasites compete within hosts and their level of competitivity is v.
 * 1) * 2002, Horst Lüddecke, "Association between Breeding Cycle and Male Body Condition in Hyla labialis," Journal of Herpetology, page 613, Vol. 36, No. 4. (Dec., 2002), pp. 607-614.,
 * Because factors like the social environment at the pond and competitivity for females may also play a role in a male's decision to stay or leave,the rules for change in individual behavior may be more complex and more plastic than those that guide the start and end of the annual reproductive cycle.

title: competitivity


text:
Talk:London Community Credit Union

External links modified
Hello fellow Wikipedians,

I have just modified one external link on London Community Credit Union. Please take a moment to review my edit. If you have any questions, or need the bot to ignore the links, or the page altogether, please visit this simple FaQ for additional information. I made the following changes:
 * Added archive https://web.archive.org/web/20150208031701/http://www.londoncu.com/general-info/what-is-a-credit-union/ to http://www.londoncu.com/general-info/what-is-a-credit-union/

Cheers.— InternetArchiveBot  (Report bug) 14:09, 5 January 2018 (UTC)

title: Talk:London Community Credit Union


text:
Sardar Raja Arts and Science College

Sardar Raja Arts and Science College, is a general degree college located in Vadakkangulam, Tirunelveli district, Tamil Nadu. The college is affiliated with Manonmaniam Sundaranar University. This college offers different courses in arts, commerce and science.

Science

 * Mathematics
 * Microbiology
 * Computer Science

Arts and Commerce

 * English
 * Commerce

Accreditation
The college is recognized by the University Grants Commission (UGC).

title: Sardar Raja Arts and Science College


text:
Talk:David Oei

Surname
Is his Chinese surname 黃, 韦, or 卫, or something else? And does he have a Chinese given (first) name? Badagnani 07:34, 16 February 2007 (UTC)

 * It's the first one. It's actually Wong spelled Oei. davidoei.weebly.com —Preceding unsigned comment added by <IP_ADDRESS> (talk) 02:27, 15 March 2009 (UTC)

Is that a Cantonese-language pronunciation? Standard Cantonese (Guangzhou dialect) is supposed to be "Wong4." Badagnani (talk) 04:16, 15 March 2009 (UTC)

 * It's from the Amoy province. —Preceding unsigned comment added by <IP_ADDRESS> (talk) 17:25, 15 March 2009 (UTC)

You mean that your ancestors came from Xiamen, in the Fujian province, before settling in Hong Kong? If so, that would mean that the language they spoke was the Amoy dialect, which is a mixture of the Quanzhou and Zhangzhou dialects of Hokkien (Min Nan). Badagnani (talk) 18:44, 15 March 2009 (UTC)

 * There is a given first name. How can one send Chinese characters? (Carlylewines (talk) 17:46, 15 March 2009 (UTC)

You may use this website to look up the characters, then copy and paste them here. http://en.wiktionary.org/wiki/Index:Chinese_radical Badagnani (talk) 18:44, 15 March 2009 (UTC)

 * My surname Oei is pronounced Wee (as in bee, see, etc.) It's the closest approximation of my native southern Fujian dialect. The biography of my great grandfather Oei Tjoe confirms this and for 14 years touring with The Intimate P.D.Q. Bach I was introduced loudly as David Wee. <IP_ADDRESS> (talk) 03:35, 19 April 2009 (UTC)

Just glanced at my history page and realized there's an editor out there deleting and disputing the pronounciation of my name. This is LUDICROUS!!!! I AM DAVID OEI AND MY NAME IS PRONOUNCED WEE. Are you telling me how my name should be pronounced? <IP_ADDRESS> (talk) 22:47, 19 April 2009 (UTC)

One of the P.D.Q. Bach albums has Peter Schickele announcing David Oei's name pronounced as "David Wee" (he further states that he holds a "black belt in piano" and that although he is from Hong Kong he has "performed on both sides of...Interstate 80." Badagnani (talk) 16:44, 20 April 2009 (UTC)

U.S. citizenship
Is he a U.S. citizen and, if so, in what year did he obtain U.S. citizenship? Badagnani (talk) 05:47, 15 December 2007 (UTC)

 * I am a citizen. Will have to look up the year. Thanks for everythig. How can I send a photo to you? davidoei.weebly.com —Preceding unsigned comment added by <IP_ADDRESS> (talk) 02:25, 15 March 2009 (UTC)

Sorry to ask such a personal question, but it helps to categorize the article (we already have you in Category:Chinese Americans from Hong Kong, which is accurate). I assume your whole family moved from Hong Kong when you came to the U.S.? Badagnani (talk) 04:14, 15 March 2009 (UTC)

 * Only my Mom came with me. Brother and Mom now live in SF. Relatives mostly in Hong Kong and around the globe. —Preceding unsigned comment added by <IP_ADDRESS> (talk) 17:29, 15 March 2009 (UTC)

Your life has been a real odyssey. Badagnani (talk) 18:50, 15 March 2009 (UTC)

Linkspam
Regarding and other edits on this same information: This is nothing more that a link to a site created by Oei's wife about herself. It's not being used as a source, but rather was added to promote her website. --Ronz (talk) 01:18, 21 April 2009 (UTC)

 * I had requested such a link only because my wife was linked erroneously to Eriko Sato the model/actress. It was only to clarify and prove that my wife was not the model/actress but indeed Eriko Sato the violinist. She's a totally successful individual and certainly needs no promotion from my article so I have absolutely no problem with this decision. That cannot be said for the other deletions made by different editors as explained further below. Finally remembered to sign my post.(<IP_ADDRESS> (talk) 16:43, 21 April 2009 (UTC))
 * I'm sure we can find a reference that meets WP:RS if this confusion needs such clarification. --Ronz (talk) 16:51, 21 April 2009 (UTC)
 * http://www.nytimes.com/1998/10/11/nyregion/music-dawn-upshaw-in-recital-at-sarah-lawrence.html mentions Oei and Sato as husband and wife. --Ronz (talk) 17:00, 21 April 2009 (UTC)

Thank you for the trouble of finding this link. It is appreciated.(Carlylewines (talk) 00:15, 22 April 2009 (UTC))
 * Glad I could help. Sorry about the drama that's occurred in response to my trying to fix this. --Ronz (talk) 00:24, 22 April 2009 (UTC)

No problem at all. The mistakes I've made on this article are only due to ignorance and computer ineptitude and not due to contempt. On the other hand it is a very unenviable position to be in as the subject of an article where truth is replaced by verifiability. I know Badagnani has no other thought than to make this a top quality article and I sincerely hope that we can all work together toward that end. Otherwise this article is useless to me and useless to Wikipedia. (Carlylewines (talk) 18:01, 23 April 2009 (UTC))

Advertisement
Regarding and other edits on this same information: Given that Oei no longer owns this shop, and no sources have been provided that show how his past ownership of this shop is an important part of Oei's life, I've removed it as overly promotional for the shop itself. --Ronz (talk) 01:21, 21 April 2009 (UTC)

 * There is no advertisement, as such is not permitted at Wikipedia. What there was was insistent and repeated deletion, quite against consensus, of important content and sourcing--something that has become a hallmark of the above editor (in addition to WP:STALK, in which he has engaged for months, and continues to engage in to this very day). Kindly cease insistently removing, against consensus, important, sourced data from our articles. The shop's ownership was clearly sourced, quite properly, by this source, which was removed again and again quite against consensus. Please restore it. Badagnani (talk) 02:14, 21 April 2009 (UTC)

 * For one thing, Badagnani, please don't engage in personal attacks and baseless threats. Second of all, directories and blatant ads don't count as sources and have no place in an article. Therefore it should stay removed. There does not have to be consensus for every single thing that is removed; rather, it should be for questionable items that are added. WP:BRD, WP:OWN and WP:DRNC apply here quite nicely.  GraYoshi2x► talk 02:42, 21 April 2009 (UTC)

 * For editors new to these problems, Requests_for_comment/Badagnani includes documentation of many similar issues. --Ronz (talk) 02:59, 21 April 2009 (UTC)

 * I'm curious how the inclusion of a store that completely occupied my life for 10 years can be considered promotional material when that store was closed FOREVER by me back in 2004. How can it be promotional material when the store no longer exist? Please explain to me what the benefits are with this PROMOTION. <IP_ADDRESS> (talk) 03:53, 21 April 2009 (UTC)
 * Maybe to promote the shop with the same name that currently exists? Who cares?  It's irrelevant.  Until we have a source that demonstrates otherwise, It's of no importance to Oei's life.  See WP:UNDUE. --Ronz (talk) 16:49, 21 April 2009 (UTC)

 * On the contrary, it is of fundamental importance to an encyclopedic coverage of this individual's life. Mentioning it in a single sentence does not promote it any more than we are trying to promote Pepsi by lovingly documenting this currently-sold beverage, produced and marketed by PepsiCo, about which we also have an article. Finally, I believe the editor insisting on removing text, as seems to be his primary interest at Wikipedia, found this article via WP:STALK, which is a very big no-no at our project, in fact against our policy. Badagnani (talk) 18:31, 21 April 2009 (UTC)

I've requested help at Neutral_point_of_view/Noticeboard --Ronz (talk) 01:18, 22 April 2009 (UTC)

This info about the store was initially written by the editor who created this article. He's responsible for nearly all the info on that page. Since I discovered the page I've made a couple of corrections and very small additions to the article, not knowing how the system works. Now that I understand the system better I shall never edit again. I did not figure out that I needed an account to do that and therefore the unsigned edits. Since I've open the account as Carlylewines I've also neglected to sign my post properly at times. That being said, I don't claim to be David Oei, I AM David Oei. 421 West 57th St. NYC<PHONE_NUMBER>. This store was opened by me on June 16th, 1994 and closed by me on June 31st, 2004. Ninety percent of my energies went into this store during that time although I managed to continue part-time in my musical life. Carlylewines didn't exist before me and it died with me. There are no other wine stores with that name that I know of. I will totally abide by the ruling on this issue and I thank user Ronz for going this route, but no one will convince me that 10 years of my life, a quarter of my adult life, is not critical to my life story. (Carlylewines (talk) 16:34, 22 April 2009 (UTC))

Sorry, the dates are Dec. 16th, 1994-June 30th, 2004. (Carlylewines (talk) 16:45, 22 April 2009 (UTC))

Carlyle Wines was chosen to be included in the inaugural issue (Nov. 1999) of 'America's Elite 1000-The Ultimate List- Millenium Issue-The Inside Story Behind America's top 1000 names' by Cadogan Publications Ltd. of UK fame. On page 80 you will find the following description of Carlyle Wines. "Kevin Bacon is a regular customer at this smart little wine store, which boasts an impressive array of wines, brandies and spirits. Each bottle has been personally selected by proprietor David Oei, whose expertise and commitment to service make this a must stop destination for wine lovers. The store is particularly strong on burgundies, which is unusual for New York. Woody Allen and Sharon Stone are also frequent customers." (Carlylewines (talk) 05:01, 23 April 2009 (UTC)) —Preceding unsigned comment added by Carlylewines (talk • contribs)

 * I found this bio from the Notice board posting. Advertisements are considered a form of self publishing so they are generally not allowed. Why not just use a better secondary source? In this NY Times article from 1999, Oei discussing the shop's preference for Campogrande. He sounds like a very hands on proprietor.--Eudemis (talk) 00:45, 24 April 2009 (UTC)
 * I'd found the same article, but decided against it because it's such a brief mention. --Ronz (talk) 20:12, 24 April 2009 (UTC)

I've gone ahead and removed the material. While it may personally be important to Oei, we have no sources showing this. --Ronz (talk) 23:02, 14 May 2009 (UTC)

Protection rights and the joys? of Wikipedia
When my friend told me I was in the Wikipedia I was stunned. I couldn't believe someone would find my life interesting enough to write an article about me. To this end I will always feel tremendously humbled and privileged. Since then the backsniping and the disputing of facts of my life have soured this honor noticeably. It completely boggles my mind that there are strangers out there deciding what my life is about and even having the unmentionable gall to decide how my name should be pronounced. My life is not something you can play around with, just as I would never do that to somebody else. It's a simple life that touched someone enough to create this article and I will forever be grateful to him. But if this continues I must request that this article be removed forever. This time I actually remembered to log in and sign. And for the last time, this is David Oei, pronounced Wee. (Carlylewines (talk) 17:02, 21 April 2009 (UTC))

 * User:Ronz, kindly stop pushing, pushing, pushing, until the subject of a WP biography, a distinguished concert pianist, no longer wishes to have his biography at our encyclopedia. The fact that this has nearly occurred means that something is very wrong. Please check yourself and think if what you are doing (insistent blanking without consensus), not only here but at dozens of pages, nearly 100 located via WP:STALK. This is simply behavior alien to our fundamentally collaborative, constructive, and collegial project. I ask in all sincerity that you contribute to our project in a manner in line with our founders' vision, always maintaining this all-important sense of collegiality and thoughtfulness, eschewing any form of aggression in one's editing. Badagnani (talk) 18:45, 21 April 2009 (UTC)
 * That's a great comment to make if you want Ronz to do the opposite of what you ask. If you want him to do what you ask, a different approach would be the intelligent choice. Your post here is not likely to produce a resolution. Do you understand? -GTBacchus(talk) 22:34, 11 May 2009 (UTC)

removing POV tag with no active discussion per Template:POV
I've removed an old neutrality tag from this page that appears to have no active discussion per the instructions at Template:POV:
 * This template is not meant to be a permanent resident on any article. Remove this template whenever:
 * There is consensus on the talkpage or the NPOV Noticeboard that the issue has been resolved
 * It is not clear what the neutrality issue is, and no satisfactory explanation has been given
 * In the absence of any discussion, or if the discussion has become dormant.

Since there's no evidence of ongoing discussion, I'm removing the tag for now. If discussion is continuing and I've failed to see it, however, please feel free to restore the template and continue to address the issues. Thanks to everybody working on this one! -- Khazar2 (talk) 00:27, 27 June 2013 (UTC)

title: Talk:David Oei


text:
Talk:Break (2020 film)

The number 1. link goes to the wrong movie. — Preceding unsigned comment added by <IP_ADDRESS> (talk) 18:08, 2 January 2024 (UTC)

title: Talk:Break (2020 film)