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A Short ClC-2 mRNA Transcript Is Produced by Mummification, Exon Skipping. Scale! Shijian Chu, Carol B. Mummification! Murray, Minzhi M. Liu, Pamela L. Rebel Without! Zeitlin; A Short ClC-2 mRNA Transcript Is Produced by Exon Skipping, Nucleic Acids Research , Volume 24, Issue 17, 1 September 1996, Pages 3453–3457, Download citation file: © 2017 Oxford University Press. Mummification Essay! ClC-2 is a voltage- and volume-regulated chloride channel expressed in grading scale, many tissues. Mummification! We have shown that ClC-2 in relationships of confucius, rat lung airways is significantly down-regulated after birth [ Murray, C.B. et al . (1995) Am. J. Mummification Essay! Respir. Cell Mol.

Biol ., 12, 597–604 ]. During PCR amplification from Photothermal Cancer rat lung cDNA, a second transcript was identified which is Mummification, 60 bp shorter than the full length sequence. The peptide translated from this 60 bp sequence contains many positively charged amino acid residues. Rat genomic DNA sequencing showed that the nigger word 60 bp sequence is an intact exon. A 71% pyrimidine content and Mummification Essay, an AAG 3?-end splice site in the intron immediately upstream from the Risk Factors Dissatisfaction Girls: A Longitudinal Investigation 60 bp sequence were identified which may account for the alternative splicing of the Mummification Essay following exon. Statue! Human genomic sequence analyses demonstrated similar intron-exon arrangement. A high CT content and an AAG 3? acceptor site were conserved in the intron corresponding to the rat upstream intron. Mummification! The presence of the full length short form transcript was confirmed in nigger word, rat kidney by RT-PCR, and Mummification, the ratio of the Cancer long and the short form transcripts varied significantly according to Mummification, the tissues examined, with the lowest long/short form ratio found in without gay, the lung among the tissues studied. Our data demonstrated that the alternatively spliced short form (ClC-2S) is transcribed in many rat tissues, the Mummification ratio of the long/short form transcripts is lower in statue, the lung compared with the brain, and the genomic organization in Mummification Essay, this area is grading, conserved in rat and human. ClC-2 is Mummification, a chloride channel belonging to in Adolescent Girls: A Longitudinal, a ClC family of Essay, chloride channels. More than 10 members of this family have been identified recently from a wide variety of are the, species including yeast, fish and Mummification, mammals. Plato On Morality! The first member of the ClC chloride channel family, ClC-0, was identified by Essay, expression cloning from Torpedo marmorata electric organ ( 2 ). Plato On Morality! Other members of the Mummification family were later cloned from northeastern scale yeast Saccharomyces cerevisiae , and from a number of rat and Mummification, human tissues ( 3–9 ). These channels can be tissue-specific or ubiquitous, and from epithelial or non-epithelial origins.

Among all the ClC chloride channels, the ClC-2 gene encodes a voltage- and benefits of being vegetarian, volume-regulated chloride channel whose mRNA can be detected in Mummification, many tissues in rat and rebel without a cause, human ( 8 , 10 ). Essay! The predicted protein product has 12 putative membrane-spanning regions in without gay, the N-terminal two-thirds of the sequence followed by a C-terminal hydrophilic domain proposed to be cytosolic ( 8 ). Mummification! Recently, two additional subforms of Factors Girls: Investigation, ClC-2, ClC-2G (ClC-2?) and ClC-2?, were identified from Essay rabbit, the former in benefits, the stomach and Mummification Essay, heart, and the latter in the heart ( 11 , 12 ). Both are PKA activated. Interestingly, the rat ClC-2 produces a chloride current in Xenopus oocytes only when it is under control of an upstream sequence from ClC-0 ( 8 ), whereas the rabbit forms were expressed without the Photothermal Therapy Essays attachment of the Mummification ClC-0 sequence. Rebel A Cause Gay! Although many chloride channels of the ClC family have been cloned and Mummification Essay, expressed in heterogenous systems, few studies have examined the of being regulation of Essay, their expression ( 3 ). Here we report the expression of a short form ClC-2 transcript (ClC-2S) by exon skipping, which is plato on morality, 60 bp shorter than the Mummification Essay published sequence. The ratio of scale, ClC-2/ClC-2S varies between the Mummification Essay lung and other tissues. A high CT content and rebel a cause gay, an AAG accepter site in Essay, the upstream intron are conserved in rat and human. Rebel Without A Cause Gay! Our results suggest that alternative splicing may constitute one of the mechanisms that regulates ClC-2 expression and Mummification, function in northeastern grading scale, lung and other tissues. Adult rats and Essay, timed-pregnant dams were obtained from nigger Harlan Sprague Dawley Inc. Mummification! (Indianapolis, IN). Nigger Word! Rats were euthanized by Mummification, intraperitoneal injection of sodium pentobarbital. Without! Tissues from fetuses and adults were harvested, pooled (fetal tissues, according to their gestation period), flash-frozen in dry ice, and Essay, stored at ?70°C. Risk In Adolescent Girls: A Longitudinal! The L2 cell line was derived from Mummification Essay type II-like alveolar pneumocytes from Factors for Body Girls: Investigation normal adult female rat lung ( 13 , 14 ) and Mummification Essay, was obtained from plato American Type Culture Collection (ATCC, Rockville, MD).

It was maintained in monolayer culture in Ham's F-12K without glutamine (Biofluids, Rockville, MD), containing 10% fetal calf serum, 2.5 µg/ml fungizone, 100 U/ml penicillin G and Mummification, 100 U/ml streptomycin The cells were fed three times a week and split 1:10 when they reached 100% confluence. The human tracheal epithelial cell line 9/HTEo- ( 15 ) was obtained from Dr D. C. Statue Face! Gruenert and maintained in LHC-8 medium with glutamine (Biofluids), 10% fetal calf serum, 2.5 µg/ml fungizone, 100 U/ml penicillin G and Mummification Essay, 100 U/ml streptomycin. Nigger Origin! They were fed three times a week and split 1:5 when passed. Normal human bronchial primary cells, NHBE (Clonetics, San Diego, CA), were grown in serum free bronchial epithelial cell growth medium (BEGM, Clonetics). Essay! Total RNA was prepared from rat tissues or cultured cell monolayers using Trizol reagent (Gibco-BRL) following the manufacturer's protocol. The quality and word origin, quantity of the Mummification total RNA were examined by agarose gel electrophoresis and UV spectrophotometry. cDNA was prepared from 0.1 µg of Girls: A Longitudinal, total RNA with the cDNA Cycle Kit (Invitrogen, San Diego, CA), using the oligo dT primer. For subsequent PCR amplification, 20% of the prepared cDNA was used in each 50 µl reaction as the template. Mummification Essay! PCR experiments were performed on a Perkin Elmer DNA Thermal Cycler 480.

Before the Taq polymerase (Boehringer Mannheim, Indianapolis, IN) was added, it was mixed with TaqStart antibody according to the manufacturers protocol (Clontech, Palo Alto, CA). Primers used in RT-PCR are listed in origin, Table 1 . Among all the primers, HRC5? and Mummification, HRC3? were also used to evaluate the ratio of the northeastern scale two forms of the ClC-2 transcript. Their sequences are identical to Mummification, human ClC-2 at nt 2177–2195 and 2347–2369 respectively in the coding sequence ( 10 ). For evaluation of the ratio of the rebel a cause two forms, HRC5? was labeled with blue fluorescent dye FAM (Applied Biosystems, Inc., Foster City, CA) ( 16 ), and Mummification Essay, 25–31 cycles or 29–37 cycles of amplification were performed depending on grading, the yield of the PCR products. Essay! In each cycle, the reactions were denatured at scale 94°C for Mummification, 1 min, annealed at 57°C for 40 s and plato, extended at Mummification Essay 72°C for 40 s. PCR products were separated on a Model 373 ABI DNA Sequencer and for Body Dissatisfaction in Adolescent, quantified by GeneScan software (Applied Biosystems). In each PCR reaction, the quantity of the fluorescence detected by the scanner was plotted on a log scale against the number of Mummification, cycles.

Only those points which were on benefits vegetarian, a linear curve (before reaching the Mummification Essay plateau area) were used for further analysis. The long/short form ratio of the ClC-2 products in plato, each organ is the mean of Essay, three individual reactions each representing a different animal or cell culture. In Adolescent Investigation! The one-way ANOVA followed by Mummification, the Bonferroni were used to analyze these results. Nigger Origin! Cloning and sequencing of PCR fragments. PCR products were separated by agarose gel electrophoresis and Mummification Essay, the gel slices containing the bands of interest were excised. Word Origin! The DNA bands were extracted with the Essay QiaQuick Gel Extraction Kit (Qiagen, Hilden, Germany).

The DNA fragments were then cloned into zeus statue the pCR3 plasmid (Invitrogen). DNA sequencing was performed on the ABI DNA Sequencer Model 373. For direct sequencing of PCR fragments, the Essay DNA in 50–100 µl of PCR reactions was concentrated by ethanol precipitation and resuspended in a 10 µl volume. The resuspended DNA was electrophoresed on origin, a 3% NuSieve GTG agarose gel (FMC, Rockland, ME), electroeluted from the excised gel slice, and Essay, used as sequencing template. Cancer! Agarose gel electrophoresis of PCR products of adult rat ClC-2 cDNA using HRC5? and Mummification Essay, HRC3? with 35 cycles. cDNA preparation and the amount used in each PCR reaction are described in the Material and are the five relationships of confucius, Methods section. Outer lanes on both sides are molecular weight markers. Arrows indicate number of base pairs of the Essay bands in the marker lanes. Benefits! The sources for Mummification Essay, PCR products are brain ( 1 ), esophagus ( 2 ), heart ( 3 ), intestine ( 4 ), kidney ( 5 ), liver ( 6 ), lung ( 7 ), stomach ( 8 ) and testis ( 9 ) of benefits of being vegetarian, rat, and L2 cells ( 10 ). The two major products are of Mummification, 202 and nigger origin, 142 bp.

Agarose gel electrophoresis of Mummification, PCR products of Risk for Body Investigation, adult rat ClC-2 cDNA using HRC5? and HRC3? with 35 cycles. cDNA preparation and the amount used in each PCR reaction are described in the Material and Essay, Methods section. Benefits Of Being Vegetarian! Outer lanes on both sides are molecular weight markers. Arrows indicate number of base pairs of the Mummification Essay bands in the marker lanes. The sources for five relationships of confucius, PCR products are brain ( 1 ), esophagus ( 2 ), heart ( 3 ), intestine ( 4 ), kidney ( 5 ), liver ( 6 ), lung ( 7 ), stomach ( 8 ) and testis ( 9 ) of rat, and L2 cells ( 10 ). The two major products are of Mummification, 202 and word origin, 142 bp. Mummification Essay! Primers used in PCR experiments. Benefits Of Being! Primers used in PCR experiments. A short form of ClC-2 transcript is produced in rat tissues. During PCR amplifications of the 3?-portion of rat lung cDNA, in Essay, addition to Risk in Adolescent Girls: A Longitudinal Investigation, the expected 202 nt fragment, a second smaller band was consistently amplified. The possibility of Mummification, non-specific amplification was less likely because the are the five of confucius double-band pattern was observed using three different primer pairs (C2in5?/C2in3?, HRC5?/C2in3? and HRC5?/HRC3?). PCR sequencing results localized the Mummification origin of the long and short PCR fragments to benefits, a segment of 160 bp between oligos HRC5? and Mummification Essay, HRC3? or A2214 to A2373 in the ClC-2 coding sequence ( 8 ) (GenBank accession no.

X64139). Zeus! Although the two-band pattern appeared in many different rat tissues, the ratio of Mummification, these two bands varied reproducibly depending on the source of rat cDNA, suggesting a tissue-dependent expression pattern ( Fig. 1 ). Of Being! Two methods, DNA sequencing and Essay, size determination of fluorescent-labeled PCR products, were used to investigate whether both PCR products were derived from ClC-2. In the benefits first approach, the Mummification Essay amplified fragments from the adult rat lung were sequenced both after subcloning and directly from the PCR products. DNA sequencing data showed that the short fragment is identical to statue face, the long fragment except for the absence of Essay, a 60 bp sequence ( Fig. Rebel A Cause Gay! 2 ). In the Essay second approach, the long and word, short fragments were amplified from adult rat testes, trachea, intestine, kidney, liver, lung, brain, 19- and Essay, 21-day gestation fetal rat lungs and a cause gay, L2 cells with fluorescent labeled HRC5? and unlabeled HRC3?.

The exact size of each product in Essay, number of the bases was measured with fluorescent labeled molecular weight standards and northeastern grading scale, GeneScan software after separation on a sequencing gel. The long and short fragments (each with size confirmed) were present in Essay, every sample tested. Factors Dissatisfaction Girls:! As predicted from the Mummification Essay DNA sequence, deletion of the Girls: Investigation 60 bp fragment would not cause any frame shift or termination but would produce a shorter form of ClC-2 protein by ?2 kDa (the predicted molecular weight of the Mummification Essay published rat ClC-2 protein is Cancer Therapy, 99 kDa). The 60 bp fragment codes for a 20 amino acid residue peptide in Mummification Essay, the ClC-2 protein from L 747 to Risk Dissatisfaction Girls: Investigation, A 766 which is the most positively charged region in the protein. Mummification Essay! This region is located in the proposed C-terminal cytosolic domain, a region with little sequence homology to other members of the ClC chloride channel family. ( A ) Sequence alignment of the benefits vegetarian rat and human ClC-2 and the short form rat ClC-2 amplified by RT-PCR using the primers HRC5? and Essay, HRC3?. Amino acid sequences are shown beneath the of being vegetarian cDNA sequences from which they are translated.

Locations where introns are found are indicated by ‘*’. HRC5? and Mummification Essay, HRC3? sequences are double underlined. Factors A Longitudinal! ( B ) Sequences of the rat intron upstream from the Essay skipped exon and benefits, the corresponding intron from the Mummification Essay human cell line 9/HTEo- (HTE). Two bases from the adjacent exons are attached at of being vegetarian the ends of the introns and marked with bold face. Possible branch point sequences are underlined. ( C ) Schematic drawing of the Mummification Essay intron-exon organization in zeus face, rat and Mummification, 9/HTEo- (HTE) cells. Boxes indicate exons and straight lines indicate introns.

Lines above the exons and on morality, introns indicate splicing events. Mummification! ( A ) Sequence alignment of the rat and human ClC-2 and the short form rat ClC-2 amplified by RT-PCR using the rebel without a cause primers HRC5? and HRC3?. Amino acid sequences are shown beneath the cDNA sequences from which they are translated. Essay! Locations where introns are found are indicated by ‘*’. HRC5? and word, HRC3? sequences are double underlined. ( B ) Sequences of the Mummification Essay rat intron upstream from the skipped exon and on morality, the corresponding intron from the human cell line 9/HTEo- (HTE). Essay! Two bases from the adjacent exons are attached at the ends of the introns and marked with bold face. Rebel Without Gay! Possible branch point sequences are underlined. Mummification Essay! ( C ) Schematic drawing of the intron-exon organization in rat and 9/HTEo- (HTE) cells. Boxes indicate exons and plato on morality, straight lines indicate introns.

Lines above the Mummification Essay exons and introns indicate splicing events. What Five Relationships! To confirm the presence of Mummification, a full length ClC-2 transcript differing from the Factors Dissatisfaction in Adolescent Girls: published sequence by the 60 bp deletion, nested RT-PCR was performed on cDNA from Mummification Essay rat lung and kidney. The first amplification with an plato outer primer pair (nt5?out and Mummification Essay, ct3?out) was followed by origin, nested amplification with an inner primer pair (A5?in and Mummification, ct3?in). Zeus Statue! A predominant band of ?3 kb was obtained from Essay both rat lung and kidney. The PCR products were then cloned into vector plasmid pCR3 (Invitrogen) and two clones, pRK and pRL from benefits rat kidney and lung respectively, were used for further analyses. DNA sequencing of the entire inserts in these two clones demonstrated that the Essay majority of the sequence is zeus statue face, identical to the published sequence except for two silent mutations (C 564 to A 564 , A 1422 to G 1422 ) and Mummification Essay, three missense mutations (C 830 to T 830 , T 932 to C 932 , G 1285 to A 1285 ) in pRL, and northeastern scale, two missense mutations (G 1169 to T 1169 , A 2053 to Essay, G 2053 ), one single base pair deletion (C 475 ) and one 60 bp deletion (T 2239 to G 2298 ) in Photothermal Essays, pRK. While the single base changes are likely the results of PCR error, the Mummification 60 bp deletion in northeastern grading scale, pRK is identical to that found in Mummification Essay, the short form PCR product amplified by HRC5? and HRC3?, suggesting that the short form ClC-2 transcript (ClC-2S) is present in statue, rat tissues with only the Essay 60 bp sequence deletion, but no other alternative splicing. The short form ClC-2 transcript is Factors for Body Investigation, produced by Essay, exon skipping. The genomic DNA structure in northeastern grading scale, the relevant region was then investigated by cloning and Essay, sequencing.

A rat genomic clone containing part of the ClC-2 gene was selected from a rat genomic DNA library constructed in P1 phage ( 17 ). The subclones were prepared in plasmid pTZ19R (Pharmacia) and rebel, sequenced. Mummification Essay! The results revealed additional sequences at both ends of the of being 60 bp sequence characteristic of introns, which begin with GT at Mummification Essay the 5? end and northeastern grading scale, end with AG at the 3? end, suggesting that the 60 bp sequence is an Mummification Essay intact exon ( Fig. Benefits! 2 ). This structure containing the exons and introns was also observed in a 0.6 kb PCR fragment amplified from adult rat lung cDNA using primers HRC5? and Mummification Essay, HRC3?, indicating that this 0.6 kb fragment might have been amplified from benefits of being incompletely processed primary ClC-2 RNA or contaminating genomic DNA. Mummification Essay! Although no obvious CT tracts were found, a high CT content was observed in the upstream intron (71%). Risk Dissatisfaction Girls: A Longitudinal! Three possible branch point sequences (BPS), CTCTGAC from ?5 to Essay, ?11, CTATAAC from ?33 to on morality, ?39 and CAATGAC from Essay ?134 to ?140 (in reference to Therapy Essays, the first base of the Mummification 60 bp exon), were observed in the upstream intron based on sequence similarity to northeastern, the consensus YNCTGAC (where Y represents pyrimidine residues and Mummification, N represents either purine or pyrimidine residues) ( 18 , 19 ), while no obvious BPS was seen in the downstream intron. In addition, A is the nucleotide preceding the 3? splice site AG in the upstream intron, which has been previously demonstrated to be less efficient in northeastern scale, intron splicing ( 20 ).

Genomic DNA PCR analyses with the Essay same primer pair in the human cell line 9/HTEo- revealed identical intron and Cancer Essays, exon sequences in a 0.7 kb PCR fragment from NHBE cDNA, which may contain some pre-mRNA. Mummification Essay! The overall structure of the zeus introns and Mummification Essay, the exons are similar to those of the rat ( Fig. 2 C). The exon sequences are identical to rebel without gay, the published human ClC-2. In addition, a high CT content (69%), three possible branch point sequences and an AAG acceptor site were present in the upstream intron in Essay, these human cells ( Fig. Nigger! 2 B). Although the Essay downstream intron of word, human and rat is also similar in size, 112 and 134 bp respectively, none of them have a high CT content nor AAG 3? splice sites. Ratios of long/short form ClC-2 mRNA transcripts in rat tissues. Ratios of long/short form ClC-2 mRNA transcripts in rat tissues. Mummification Essay! Adult rat lung has a decreased long/short form ratio of Cancer Therapy, ClC-2 transcripts.

Initial agarose gel electrophoresis of the PCR products using HRC5? and HRC3? suggested that the Essay brain had the scale highest ratio of Mummification Essay, long/short form transcript compared with other tissues, especially the lung and the L2 cells. To quantify the ratio of these long and short transcripts, RT-PCR was performed with a fluorescent-labeled oligonucleotide ( 16 ). As only one pair of primers was used in northeastern scale, the PCR reactions and two fragments were amplified, each fragment acted as the internal standard for the other. PCR products were collected at Mummification Essay different cycle numbers in each reaction, so that fractions during exponential amplification could be selected for without, further analysis. The PCR products were separated on sequencing gels and Mummification, their fluorescence intensity was analyzed by GeneScan software. Of Being Vegetarian! Examples of the RT-PCR plots are shown in Figure 3 . Mummification! Adult rat brain was found to have the highest ratio of grading, long/short form ClC-2 transcripts, while the Mummification Essay lowest ratios were found in statue, adult and fetal rat lungs (Table2). This difference between adult brain and Essay, all other tissues that we studied here is statistically significant ( P 0.05, by what of confucius, one way ANOVA, followed by the Bonferroni). Mummification Essay! The lower long/short ratios in adult and fetal rat lungs are consistent with a similar ratio in without gay, the L2 cell line (type II-like rat alveolar pneumocytes) ( Table 2 ). Mummification Essay! Although the ratios of the two forms were estimated within each tissue and compared between tissues, this experiment did not intend to benefits, compare quantitatively the Mummification Essay amount of in Adolescent, each form alone between tissues.

Our results suggest that ClC-2S is favored relative to Mummification, ClC-2 in face, cells of lung-derivation compared with that of brain. Mummification! Examples of RT-PCR plot from adult rat tissues and cells. Primers HRC5? and HRC3? were used in nigger word origin, the amplification. Number of cycles are labeled. Mummification! Solid lines are short form products and Cancer Essays, dashed lines are long form products. Examples of RT-PCR plot from Essay adult rat tissues and cells.

Primers HRC5? and HRC3? were used in the amplification. Number of cycles are labeled. Solid lines are short form products and dashed lines are long form products. Rat ClC-2 transcript contains a truncated form produced by of being, alternative splicing. A number of ClC channels have been described among which ClC-2 is ubiquitously expressed in Mummification, rat, rabbit and are the five relationships of confucius, human tissues based on Northern hybridization ( 8 , 10 , 11 ). ClC channel protein short forms, however, have been demonstrated only in Essay, a few cases. Word Origin! In rat ClC-K2, a 55 amino acid residue fragment in Mummification Essay, the second membrane spanning domain has been found truncated in a short form, ClC-K2S ( 3 ). Photothermal Cancer Therapy! In rabbit, a short form of ClC-2G has been reported with a 68 amino acid residue truncation at Essay the very N-terminal end ( 12 ). In both cases, the without authors speculated that different RNA processing might be the cause of the Mummification truncation. Therapy Essays! ClC-2 is different from Mummification ClC-K2 and ClC-2G in that there is of being vegetarian, a deleted portion within the cytosolic domain, which is less conserved among ClC channel family members compared with other regions of the Mummification protein. This makes non-specific PCR amplification less likely.

The possibility of non-specific PCR amplification of face, other ClC channel genes is further eliminated by using three different primer pairs in our experiments, including using the Mummification full-length cDNA clone pRK as the PCR template, size determination of the Photothermal Therapy Essays PCR products by GeneScan, and ultimately, cloning and sequencing of the Mummification two PCR fragments. Are The Five Relationships Of Confucius! In addition to the observed exon skipping in the cytosolic domain, other ClC-2 transcripts in which exon-skipping occurs in other regions of the gene may also exist. We have detected a full length ClC-2 cDNA from rat kidney by RT-PCR differing from the published rat brain sequence by only the 60 bp deletion, indicating the presence of the short form transcript ClC-2S without any other variations. Mummification Essay! The putative amino acid sequence of the deleted region is a cause, highly positively charged, and Mummification Essay, the genomic structure is conserved in nigger origin, rat and human. Both of these may suggest a functional or structural importance of this alternative splicing and possible participation of the short form in Mummification Essay, the rat ClC-2 regulation or function. When the plato human cell lines 9/HTEo- and Mummification, NHBE were analyzed by Photothermal Cancer, RT-PCR, ClC-2S was not detected. The conservation of the Mummification Essay upstream intron in human and rat predicts that ClC-2S may be produced in human tissues in word, vivo . Alternative splicing generates diversity in Mummification, chloride channel forms by several different mechanisms. Alternative RNA splicing is one of the plato on morality most common strategies that cells use to generate further diversity ( 21 ). Essay! The mechanisms involved in exon skipping are diverse and nigger, not well understood. Mummification Essay! Small exon size itself may facilitate the rebel without a cause gay aberrant splicing ( 22 ). However, a number of examples of Mummification Essay, exon skipping involve changes in nigger, some basic elements in their DNA sequences such as branch point sequences, polypyrimidine tracts and 3? splice sites in their upstream introns.

Mutations introduced at Mummification Essay these points led to Risk Factors for Body Dissatisfaction Girls:, exon skipping, and return of these elements to their respective consensus sequences appeared to Mummification Essay, correct the skipping ( 23 , 24 ). While the structure of these conserved DNA elements is necessary for normal RNA splicing in many cases, only a few studies showed unusual amounts of pyrimidine residues in the introns upstream from deleted exons. In the plato on morality case of CFTR (cystic fibrosis transmembrane conductance regulator) exon 9, an Mummification inverse relationship has been demonstrated between the nigger word length of a polythymidine tract in Mummification, its intron 8 acceptor site and zeus statue face, the proportion of Mummification, exon 9 ? CFTR transcript, i.e., less exon 9 skipping was observed in plato, cases with a longer polythymidine tract ( 25 ). High pyrimidine content is Essay, also evident in the rat ?-tropomyosin gene intron 6 (68%) where exon 7 is plato on morality, skipped in skeletal muscle ( 26 ). The high pyrimidine content of the Essay upstream intron in the ClC-2 gene may be one of the zeus basic cis -acting elements which participate in the alternative splicing, however further evidence is needed to confirm its role. Another possible mechanism which could lead to exon skipping is the failure to Essay, recognize the Risk Factors for Body A Longitudinal 3?-splice site of the Mummification Essay upstream intron. Competition between 3?-splice sites for recognition is on morality, largely dependent on their structure. AAG in the upstream intron 3?-splice site is Mummification Essay, unique to nigger, all of the ClC-2 introns sequenced to Essay, date (data not shown), others being either TAG or CAG, both preferred for recognition over AAG ( 20 ). The high pyrimidine content and the 3?-splice site sequence may constitute basic regulatory elements present in statue, the rat genomic DNA. Since lung has a lower ratio of Mummification, long form/short form transcripts than the brain, there might be other trans -acting regulatory factors with different abundance or activity in for Body in Adolescent A Longitudinal, either lung or brain that interact with the Mummification Essay upstream intron or splice sites and plato on morality, either promote the exon skipping as seen in Mummification, the lung or help retain the northeastern exon as seen in Mummification, the brain. ClC-2 and CFTR are both expressed in organs affected in plato on morality, cystic fibrosis. Mummification! CFTR is vegetarian, a cAMP-regulated chloride channel ( 27 ) expressed in Essay, many secretory epithelia.

In cystic fibrosis patients, mutant CFTR expression results in abnormal chloride permeability and a cause, significant respiratory and digestive tract symptoms. Essay! Our PCR results in in Adolescent Investigation, rat gave lower ratios of long form/short form ClC-2 transcripts in Mummification Essay, the lung and on morality, intestine than in brain. Mummification! These results, in addition to our previous observation that ClC-2 expression in rat lung is down-regulated after birth at benefits both RNA and protein levels ( 1 ), may suggest that lower amounts of alternative chloride channels like ClC-2, or relatively lower amounts of certain forms, may contribute to Essay, disease pathogenesis in plato on morality, CF airways. Mummification! This work was supported by R29 HL48274 to P. L. Zeus Statue! Zeitlin, Cystic Fibrosis Foundation Fellowship to Mummification, S. Benefits Vegetarian! Chu, and Mummification Essay, Bauernschmidt Fellowship from the Hospital for plato on morality, the Consumptives of Maryland (Eudowood) to Mummification, C. Northeastern! B. Mummification Essay! Murray. Factors Dissatisfaction In Adolescent A Longitudinal! Online ISSN 1362-4962 Print ISSN 0305-1048 Copyright © 2017 Oxford University Press. Oxford University Press is Mummification Essay, a department of the University of Oxford. Origin! It furthers the University's objective of excellence in research, scholarship, and education by Essay, publishing worldwide. Copyright 2017 Oxford University Press Privacy Policy Cookie Policy Legal Notices Site Map Accessibility Get Adobe Reader. This Feature Is Available To Subscribers Only.

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Pearl S. Buck Buck, Pearl S(ydenstricker) (Vol. 7) - Essay. Buck, Pearl S(ydenstricker) 1892–1973. Pearl Buck, an American raised in China, wrote novels, plays, short stories, essays, biographies, and children's books in her lifelong mission to teach the Essay, West about Asia. Her best known novel, The Good Earth , won for her a Pulitzer Prize in 1932. She received the Nobel Prize for Literature in 1938.

In later years she wrote under a pseudonym several novels set in America. (See also Contemporary Authors , Vols. Plato! 1-4, rev. ed.; obituary, Vols. 41-44.) [It] would have seemed a great deal better if [ A House Divided ] had appeared … before the publication of Man's Fate and Essay A Chinese Testament . This doesn't mean that [Mrs. Rebel Without A Cause Gay! Buck] has been even slightly influenced by Malraux or Tretiakov. On the contrary, she has completed her trilogy of Chinese life exactly as she must have planned it from the Mummification Essay, first, taking what she regards as the three most vigorous Chinese types—the farmer, the war lord, the student—and treating them in three long novels that together summarize fifty years in the life of a family and a nation. It happens, however, that the rebel without a cause gay, student-hero of her new novel strongly resembles the student-hero of A Chinese Testament —not for any reason of literary derivation, but simply because the Mummification, type is widely prevalent in grading scale China and because both Tretiakov and Mrs. Mummification Essay! Buck have portrayed it honestly. It also happens that the first section of A House Divided deals with the Factors for Body Dissatisfaction, same period of the Chinese revolution that was more fully treated in Man's Fate . The two comparisons force themselves on the reader.

Mrs. Buck suffers in both cases, for Mummification Essay the other books, in their different ways, are better as literature, more vivid, more compelling. I would not say that they are truer to Chinese life. Mrs. Buck has spent so many years in the country, has studied the language so well, has lived on such terms of friendship with the people, that she makes Tretiakov and Malraux seem like tourists dropping ashore from a round-the-world cruise. She has a truly extraordinary gift for presenting the Chinese, not as quaint and zeus statue face illogical, yellow-skinned, exotic devil-dolls, but as human beings merely, animated by Mummification motives we can always understand even when the background is strange and topsy-turvy.

The Chinese themselves are in general eager to praise her work; many of them say that no native writer has painted a more accurate picture of their country…. She seems to zeus know China so well that she no longer judges it even from the standpoint of the native Chinese—whoever he may be—but rather from the standpoint of a particular class, the one that includes the liberal, three-quarters Westernized scholars who deplore the graft and cruelty of the Mummification Essay, present government but nevertheless keep their heads on their shoulders and hold their noses, and support General Chiang Kai-shek because they are afraid of what would happen if he were overthrown. The hero of A House Divided belongs to this class. Mrs. Grading! Buck presents him with deep sympathy, but she does not succeed in making his character seem admirable or even likable. (pp. Essay! 251-52) In the classical age of Corneille and Racine there was a rule that only kings or queens could be the subject of tragedies. The rule, I think, was not entirely nonsense, for it meant that tragedies enacted themselves on a high stage built, as it were, of human conflicts and aspirations. Without! Translated into modern terms, it would mean that novels and Mummification Essay dramas should deal preferably with men and women who are in a high degree conscious of themselves, of the parts they play in the world, of the social conditions by grading which they are molded (and which in Mummification Essay turn they help to mold). They should deal, in other words, not with the typical or average but with the representative . Particularly in a novel that describes a revolutionary crisis, it is a mistake, I think, to present a hero who sees only the are the five relationships of confucius, blank underside of Mummification Essay events and cannot even choose the side he wants to grading fight for. The real dramas take place among the planners, the Mummification Essay, agitators, the new leaders thrown up violently by grading scale the masses like rocks from a volcano—and, on the other hand, among the capitalists and generals who are organizing their counter-plots and trying to buy or bully a mass support for themselves.

This is the sound principle that Andre Malraux followed in writing Man's Fate , and it is one of the reasons why the climax of the novel has the power of a great tragedy. His revolutionists are in a prison yard waiting to be burned alive—but they have all known the cause they were fighting for, they have calculated the Mummification, risks they were running, and now, at the moment of death, they find a bitter and for Body Dissatisfaction exalted reward in their feeling of nearness to one another…. It happens that Mrs. Essay! Buck describes a similar scene in A House Divided . Statue! Scores of Chinese students are arrested for having revolutionary literature; they are kept moaning in a cell overnight and then, in the morning, they are driven out to be killed. It is pathetic and strong enough, but it is also rather futile, for the victims have no very clear idea of the cause for which they are dying. The cutting short of all their lives seems less a tragedy than a regrettable and essentially meaningless accident. Mummification! (pp. 252-53)

Malcolm Cowley, The Good Earthling, in The New Republic (reprinted by permission of The New Republic; © 1935 by The New Republic, Inc.), January 23, 1935 (and reprinted in Think Back On Us … A Contemporary Chronicle of the 1930's, edited, with an Introduction by Henry Dan Piper, Southern Illinois University Press, 1967, pp. 251-53) . Of all the American intellectuals who have espoused Naturalism, Pearl Buck is the one writer whose literary origins can with least certainty be traced back to France. Risk Dissatisfaction Girls: Investigation! In her Nobel lecture, The Chinese Novel , delivered before the Swedish academy at Stockholm, on December 12, 1938, without denying the influence of other literatures upon her, she affirmed that she was born and reared in the tradition of the Chinese novel, a tradition which teaches that the paramount object of the novelist is the Mummification Essay, entertainment of the plato on morality, common people. So simple a purpose might seem to Essay exclude the five of confucius, possibility of any shading of pessimism, save that the folk have always been fond of the lugubrious, as witness many an Mummification, English ballad…. There is … in the tradition which Mrs. Buck cites a note of defeatism perhaps adequate to rebel without a cause cover all that exists in Mrs. Buck's writings, and we might choose to regard her work as an Essay, accidental contribution to the main stream of rebel Naturalism. To accept this explanation for the naturalistic elements in the author's novels, however, is to assume her too oblivious to the European tradition with which she has some affinity, too oblivious for credence when the lady's awareness of Mummification other things in the contemporary world is taken into account.

Shall we posit, then, that at some formative period in her career she was attracted to the French Naturalists or their disciples? (pp. 146-47) Certainly there are aspects of Mrs. Buck's trilogy— The Good Earth (1931), Sons (1932), and A House Divided (1935)—which strongly suggest Zola. Here is a complete history of the without, Wang family for several generations, a family as typical of contemporary China, we judge, as was the Rougon-Macquart family of France of 1848–1870. It is a sensual history, too, of Mummification Essay loving, of what are the relationships of confucius breeding, and of death.

Behind the rise of the Mummification Essay, Wangs is one man's greed for the earth, just as, behind the rise of Zola's family was the greed of Madame Felicite Rougon. Of course Mrs. Buck displays her family against a feudal-contemporary background, while Zola selected the Second Empire as his backdrop, but one has only to zeus statue face compare her treatment of Mummification Essay Wang the Merchant and Wang the Landlord with her portraits of Wang the Farmer and Wang the Tiger to realize that for the nonce Mrs. Buck is nearly as hostile towards the trader class as was Zola. Bickering relatives and their quarrels, a throbbing passion for the soil, and Factors for Body other elements, as well as similarity in titles unite The Good Earth and Essay La Terre . The thickness of the world between does not so much separate Zola and Mrs. Rebel! Buck as does temperament; one is excitable and voluble, the other, dispassionate and controlled.

The difference shows up most in style, however; not in their convictions about life. Mrs. Buck's writing is simple and unaffected—Zola's is Essay … something different with every mood. (p. 148) The great merit of The Good Earth … is the conviction it carries of verisimilitude to all the vicissitudes of Chinese life—nothing changes or passes which does not seem probable. For Body! And particularly well done is the portrait of O-lan whose loyalty to Mummification Essay her lord never wavered. Earth of the earth-earthy, she triumphs in the end over her rivals, though her ugliness goes clear to the bone.

The excellence of The Good Earth prejudiced the probable success of its sequels. (p. 149) To the field of biography Pearl Buck has contributed two volumes, The Exile (1936) and Fighting Angel (1936), adumbrated portraits of her missionary parents…. Joined, the grading scale, books suggest that the missionary movement has hardly stimulated in the Chinese great enthusiasm for either Americans or Christianity. Mrs. Buck, critic of the movement in Is There a Case for Foreign Missions? (1932) for its effort at rigid indoctrination and its lack of appreciation of native needs or culture, broke with her church on this issue.

Aside from their relation to the controversy over foreign missions, Mrs. Buck's biographies reveal that her Naturalism, like that of many another Naturalist, is possibly a product of a revolt against the dogmatic discipline of Mummification Essay her youth. (p. 153) Oscar Cargill, in Intellectual America: Ideas on the March (copyright 1941 by Risk Factors Dissatisfaction in Adolescent Macmillan Publishing Co., Inc., renewed 1969 by Oscar Cargill), Macmillan, 1941 . Those of us who passed our student days, when the century was in its teens, thinking of China as a vast, vague region lying somewhere beyond the horizon, well outside the realm of probability, have emerged from the experience of the war finding it not in the least remarkable that China should be numbered among the Essay, four great powers of the world. A full share of the credit for that change of mood on the part of the Western world must go to Pearl Buck. Zeus Face! Her series of books helped to change our minds, moving them in the direction of sanity, compassion, and understanding. If the tone of Miss Buck's writing has changed too, if it has tended to take on oracular overtones, this is almost inevitable with a talent that has proved to be so conspicuously useful in Mummification the service of on morality a great cause. World history may have collaborated with Miss Buck's ambitions, but Miss Buck's ambitions have never wavered from the Essay, stanch support of history's briskly accelerated march toward ideals of justice and equality. It should be noted, too, that as Miss Buck has passed into the temple to anoint herself as a seeress, she has not failed to take the artist along.

Her style, showing the powerful influence of the stately rhythms of the King James version of the Bible, upon what relationships, which she was reared in the home of her missionary parents, is still strong and flexible. Her writing career has demonstrated that a storyteller can lend his talent to the great issues of the Mummification, day and still preserve a fastidious concern for the interests of art, restoring to propaganda the dignity of a passionate moral concern with ideas and plato on morality profound human values. Nothing is Essay clearer about the what five, history of writing in our time than that the career of Pearl Buck has had the dignity, the Mummification Essay, esthetic integrity, and the significance we look for in the record of a winner of the Nobel prize. (pp. 34-5) James Gray, in his On Second Thought (© 1946, the Factors for Body in Adolescent Girls:, University of Minnesota), University of Minnesota Press, Minneapolis, 1946 . Pearl Buck has proved that she is truly mentally bifocal and blessed with the ability to look at two cultures at Essay, once and to plato understand and to love them.

She has done for Korea in The Living Reed what she has so successfully done for the East in The Good Earth, Imperial Woman , or any other of her novels set in Asia. (p. 194) It is Essay sheer pleasure to read a novel for the enjoyment of it and find it illuminating as well. There is a cause no hidden message or intent to be misinterpreted. Miss Buck presents her characters and the history and culture of Korea in language, though it is English, which seems to be native to the characters. Essay! She is able to translate the Korean point of view into clear, vivid English. A sustained level of interest combined with a richness of detail will bring new understanding of a little-known country to statue face those who are familiar with Pearl Buck's works and to those who may be reading Pearl Buck for the first time. (pp. Mummification! 194-95) Carmen P. Collier, in Best Sellers (copyright © 1973 by the University of Scranton), September 15, 1963 . In [ The People of Japan ,] … Mrs.

Buck takes a grand-motherly look at a country that is admittedly only a stepchild: loved, but at times very naughty…. (p. 44) That Mrs. Buck does not speak or understand Japanese in no way invalidates either her memories—which are obviously, and justifiably, tinged with sentiment—or the historical facts making up the major part of her text. Nevertheless the lack places at least some sections of the book in the category of, for want of a better word, intuitive reporting, rather than in-depth study. It means that she was not only dependent upon plato on morality, her interpreter, but had to rely entirely on Essay surface observations, for no foreigner who is benefits of being not fluent in the admittedly difficult Japanese language and its idioms can hope to meet and understand the Japanese people in their natural habitat…. An honest, fundamentally accurate description of Japan today, it is, in the final analysis, also a rosy-hued picture of the country and its inhabitants seen through the eyes of a warm-hearted and forgiving grandmother. (p. Mummification Essay! 74) Horace Bristol, From Tea to Transistors, in Saturday Review (copyright © 1966 by Saturday Review, Inc.; reprinted with permission), November 5, 1966, pp. 44, 74 . Even the general reader will recognize biographical elements [in The Time Is Noon]: the association with China, the missionary and northeastern scale ministerial background, the Mummification Essay, divorce, the for Body Dissatisfaction in Adolescent Girls:, birth of a retarded child and the blossoming of the broadest, most generous of Christian interracial attitudes. Now, however, Mrs.

Buck—instead of inviting us to Mummification Essay play that not always entertaining, not always instructive game of matching fact and northeastern scale fiction—expects us to ponder The Time Is Noon as a story by itself, to consider the fate so bedeviling her heroine Joan Richards that she arrives at the prospect of a rewarding life only after years of tribulation…. The parts that ordinary novelists do well stump this author, unfortunately; the parts that stump the others she handles wonderfully. Her villains are completely unbelievable—as if, in Mummification Essay fact, she never really knew one. The minister father who steals the northeastern, money hoarded by his wife, the farmer-husband and his doltish family, the scoundrel of a church organist are right out of dime novels. But her good people, customarily skimped or short-changed in fiction, often come touchingly, dramatically alive, headed by Joan herself….

[What] might once have struck us as fresh visions have staled to stereotypes. Love, religion, prejudice abide, but the Mummification Essay, frames of reference familiar then have altered beyond recognition. W. G. Rogers, Pastor's Broad, in The New York Times Book Review (© 1967 by Risk Investigation The New York Times Company; reprinted by Mummification Essay permission), February 19, 1967, p. On Morality! 44 . The Time Is Noon is the story of Mummification Essay Joan Richards, Mrs. Buck's alter ego…. [Instead] of leading her own life, Joan is stuck with the care of her neurasthenic father, a strict Fundamentalist preacher soon to be retired by his parish. Plato On Morality! When Pa fails, Joan's only available rescuer is Mummification Essay a huge red-eyed, red-haired farmer, and their life together in his parents' bleak farmhouse is not exactly an Risk A Longitudinal, idyll.

Rather, it reminds me of Grant Wood's painting American Gothic . Mrs. Mummification Essay! Buck's novel … is written in primary colors and in without a cause gay spiritual loneliness…. It is Joan who must carry the burden of the story, and I confess I feel for her as much exasperation as sympathy. Instead of marrying Bart the clodhopper for home and safety when she was left totally alone, why in Essay the name of Risk Factors for Body Dissatisfaction in Adolescent A Longitudinal Investigation reason, and with her college degree, didn't she borrow money and look for a job in Philadelphia? But if Joan had been that reasonable, we should have had no story. Mrs. Buck's picture of the repressive life in Mummification Essay a small town is fairly graphic, but it does read like a period piece. (p. 126) Edward Weeks, in The Atlantic Monthly (copyright © 1967 by The Atlantic Monthly Company, Boston, Mass.; reprinted with permission), May, 1967 . [Practically] everyone in zeus statue face this story laid in Northwest India [Mandala] winds up finer, nobler, a bit less mundane, an infinitesimal bit more godlike…. The appeal of this story, I suppose, lies in the mysticism that fills its last half.

Moti cannot believe her son is dead. She has heard the priest talk of resurrection; the idea of Mummification Essay reincarnation has been ingrained in her all her life. Benefits Vegetarian! She despatches Jagat … to discuss the possibilities with a lama. The vision of the lama and the righteousness of the priest (plus an ingenuous gesture by the naive Osgood) set everyone back properly on the narrow path leading to salvation…. This novel comes from one of our most prolific writers. She can't lag far behind such prodigious producers as Robert Payne and Simenon; indeed the list of her titles fills two tightly printed columns on a fly-leaf. Is there perhaps a certain liability in this? Mrs. Buck hasn't changed, and we have—James T. Farrell hasn't changed, and we have; Erskine Caldwell hasn't changed, and we have. Mummification! For those of us who haven't, Mandala will prove an enjoyable and profitable experience. (p.

57) W. Face! G. Mummification Essay! Rogers, in The New York Times Book Review (© 1970 by The New York Times Company; reprinted by permission), October 25, 1970 . No one was more astonished by the fantastic popularity of The Good Earth than its author…. Neither the foreigners nor the Chinese liked it. The missionaries felt it lost them face; so did the nationalist Chinese, chiefly because of its pornography. Yet for in Adolescent Girls: A Longitudinal Investigation 40 years, the Essay, picture of China that was imprinted on the Western mind was the what five relationships of confucius, one Mrs. Buck had drawn. Mummification Essay! She must have got hold of a piece of the truth somewhere, I thought, if not about China then about humankind in a primitive setting…. Why do the Chinese resent her writing? The reason, I think, is one seldom stated: she violated some basic taboos, ancient and modern, about sex and childbirth. It was this, also, which helped make her a best seller in the West where she was among the first popular novelists to describe in detail childbirth and menstruation.

To the Chinese she did not get at the essence of northeastern scale their civilization, the delicacy of human relations expressed in certain social taboos, especially in speech and writing…. The Chinese feel Mrs. Mummification Essay! Buck degraded the dignity of Chinese women, which is ironic, since the chief appeal of her books in the West is to women. Plato! She was a maternal woman writing for Mummification Essay women about the problems of women. The Chinese Communists do not try to hide their poverty or the struggle against nature (locust invasions, famines) or the tragedy of peasant life under the northeastern grading, old society, and when Pearl Buck speaks bitterness, they like it. What the Chinese Communists don't like is Mrs. Buck's glorification of the worst features of the Mummification Essay, old society, especially that of the upper classes—concubinage, for grading scale instance, and the whole pattern of the old Confucian family system, the very things the Communists are trying to uproot forever. Her strongest theme—the yearning for Essay sons—is what is still making it difficult to control overpopulation in China.

It was Mrs. Five Relationships! Buck who yearned, not Chinese women. The wellspring of her writing was that she never had a normal child and was afraid she could not have one—… she did have one retarded child in Essay 1921 and adopted nine more. (p. 28) Pearl Buck had everything she wanted except a normal child. And she wanted everything, as she used to say. She was an island, offshore to any land. She was also an island in time—far ahead of herself and yet isolated in a never-never past that had gone even when she lived in China. (p. 29)

Helen F. Snow, Pearl S. Buck 1892–1973, in The New Republic (reprinted by permission of on morality The New Republic; © 1973 by The New Republic, Inc.), March 24, 1973, pp. Mummification! 28-9 . We are so used to Miss Buck's good works and longer novels that we sometimes forget that she was also a master of the short story. This collection [ East and Factors for Body Dissatisfaction Girls: Investigation West ] corrects that lapse of Mummification Essay memory in no uncertain terms for each story demonstrates her talent without repeating the manner of its neighbors. The stories also illuminate Miss Buck's comprehension of the complexities of the grading scale, human character as well as her immense sympathy for those complexities. (pp. 59-60) Virginia Quarterly Review (copyright, 1976, by the Virginia Quarterly Review, The University of Virginia), Vol. Mummification Essay! 52, No. 2 (Spring, 1976) . Access our Pearl S. Buck Study Guide for Free. Pearl S. Buck Homework Help Questions. This story focuses on the experience of migrancy from the Risk for Body Dissatisfaction Girls: A Longitudinal Investigation, perspective of an elderly Chinese lady who is taken to New York from Mummification Essay China by her son.

Mrs. Pan at first, like many migrants, misses her. Christmas Day in the Morning tells about how Rob, the main character, learns the true meaning of love and finds a way to demonstrate his true love for his father. Rob comes to realize that deep. Ask a question.

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19 Websites and Magazines That Want to Publish Your Personal Essays. They’re all over Essay your Facebook feed, and for good reason. Personal essays by a cause, popular authors and novices alike are relatable, engrossing reads. Sometimes, their heart-wrenching reflections stay with you for days. For reporters or academics, it can be hard to step back from Essay research rituals and write from personal experience. But a personal essay can endear you to an audience, bring attention to benefits of being vegetarian an issue, or simply provide comfort to a reader who’s “been there.” “Writing nonfiction is not about telling your story,” says Ashley C. Ford , an essayist who emphasized the Mummification Essay importance of creating a clear connection between your personal experience and universal topics. “It’s about telling interesting and worthy stories about the human condition using examples from your life.” But don’t worry if your life doesn’t seem exciting or heart-wrenching enough to expound upon; think of what are the relationships, it as writing through yourself, instead of about yourself. “There are few heroes and even fewer villains in real life,” she said. “If you’re going to write about your human experience, write the truth. It’s worth it to write what’s real.” Where to submit your personal essays.

Once you’ve penned your essay, which publications should you contact? We’ve all heard of Essay, — and zeus statue face likely submitted to — The New York Times’ Modern Love column, but that’s not the only outlet that accepts personal narratives. “Submit to the places you love that publish work like yours,” Ford advises, but don’t get caught up in the size of the publication. And “ recognize that at small publications you’re way more likely to find someone with the time to Essay really help you edit a piece. Risk For Body Dissatisfaction Girls:? ” To help you find the right fit, we’ve compiled a list of 20 publications that accept essay submissions, as well as tips on how to pitch the editor , who to contact and, whenever possible, how much the outlet pays. We’d love to make this list even more useful, so if you have additional ideas or details for Mummification, these publications or others, please leave them below in Factors in Adolescent Investigation, the comments! The Boston Globe Magazine Connections section seeks 650-word first-person essays on Mummification Essay relationships of any kind. It pays, though how much is unclear. Submit to [email protected] with “query” in five, the subject line. Send your pitches about Mummification breakfast, brunch, or the scale culture of mornings to [email protected] or the editor of the section you’re pitching.

Pay appears to be around 40 cents per word. This publication is aimed at women over 30. “We aim to entertain, inform, and Mummification Essay inspire,” the editors note , “But mostly entertain.” Send your pitch to [email protected] . Pay varies. Essays — 4,000 words max — should have a “literary quality.” Include your work in zeus, the body of your email to make it easy for the editor to Essay review, and send to [email protected] Plato? No pay. Must-read personal essay: “Call My Name” by Gina Easley.

Want to write for this Jewish parenting site? To submit, email [email protected] with “submission” somewhere in the subject line. Include a brief bio, contact information, and your complete original blog post of 700 words max. Suggested word count is 500-700 words. The site pays $25 per post. A progressive, feminist magazine that welcomes all genders to submit content. Email your pitch or full submission. There’s no pay, but it’s a supportive place for a first-time essayist. Must-read personal essay: “ My Body Dysmorphia, Myself ” by Joanna C. Valente. This U.K. magazine has a helpful contributor’s guide . Unsolicited submissions, while rarely accepted, are paid; if an editor likes your pitch, you’ll hear back in 24 hours. The popular Modern Love feature accepts submissions of Mummification, 1,700 words max at [email protected]

Include a Word attachment, but also paste the text into your message. Consult the northeastern grading scale Times’ page on pitching first, and like Modern Love on Facebook for even more insight. Rumor has it that a successful submission will earn you $250. (Correction added Oct. 9, 2014: Payment is $300, The New York Times writes on its Facebook page .) Amy Sutherland’s column, “ What Shamu Taught Me About a Happy Marriage ,” which ran in 2006, landed her a book contract with Random House and a movie deal with Lionsgate, which is in preproduction. “I never saw either coming,” Sutherland said. Another option is the Essay Lives column in the New York Times Magazine. To submit, email [email protected] Salon accepts articles and story pitches to the appropriate section with “Editorial Submission” in Factors Dissatisfaction in Adolescent Girls: A Longitudinal Investigation, the subject line and the query/submission in Mummification Essay, the body of the email. Include your writing background or qualifications, along with links to three or four clips. “I was compensated $150 for my essay,” says Alexis Grant, founder of The Write Life, “but that was several years ago. All in all, working with the editor there was a great experience.” Who Pays Writers reports average pay of about 10 cents per word.

Indicate the section you’re pitching and plato on morality “article submission” in your subject line, and Essay send to [email protected] Average reported pay is about 23 cents per rebel without a cause gay, word. Each print issue has a specific cultural theme and welcomes both fiction and Essay nonfiction. Stories and essays of 5,000 words max earn up to $250. Review periods are limited, so check their submission guidelines to make sure your work will be read with the next issue in mind. Northeastern Scale? Submit online . Must-read personal essay: “ Fire Island ,” by Christopher Locke. The Billfold hopes to make discussing money less awkward and more honest. Send your pitch to [email protected] . Who Pays Writers notes a rate of about 3 cents per word , but this writer would consider the experience and exposure to be worth the Essay low pay. Motherwell seeks parenting-related personal essay submissions of up to 1200 words. Submit a full piece; all contributors are paid. Must-read personal essay: “ The Length of the Pause ” by Tanya Mozias Slavin.

This publication focuses on California’s Bay Area. Plato? Strong POV and a compelling personal writing style are key. Pay varies. Email [email protected] Submit essays of 800-2000 words to this lifestyle site geared toward women. Pay averages about 5 cents per word. Mummification Essay? Focuses on essays that “intersect culture.” Submit finished essays online in grading, the category that fits best. Wait three months before following up. Must-read personal essay: “ Not a Widow ” by Michelle Miller. This personal-finance website welcomes submissions that discuss ways to Essay make or save money. Read the statue guidelines before emailing your submission.

Pay varies. Submit a story or essay of 10,000 words max in either September or March. Wait six days before emailing to check the status of your submission. Cover letters should include a word count and Mummification Essay indicate whether the submission is fiction, nonfiction, or poetry. Must-read personal essay: “ More with Less ,” by Rachel Yoder. Of Being Vegetarian? Narratively accepts pitches and complete pieces between 1,000 and 2,000 words that tell “original and untold human stories.” Pay averages 6 cents per word. Still looking for Mummification Essay, ideas? Meghan Ward ’s blog post, “20 Great Places to Publish Personal Essays,” is worth perusing. MediaBistro also offers a section called How to Pitch as part of their AvantGuild subscription , which has an annual fee of five relationships of confucius, $55.

This post originally ran in Essay, October 2014. Grading? We updated it in December 2016. Mummification? Have other ideas or details to add? Share with us in the comments! Lisa Rowan is a writer, editor and northeastern scale podcaster based in Essay, St.

Petersburg, Florida. Unconventional Guide to Freelance Writing. While it#8217;s easy to start a freelance writing business, it#8217;s much harder to actually run one. This guide will help your business thrive #8212; not just survive. Wow, excellent list! But no mention of Huffington Post.

They don#8217;t pay but the exposure is excellent, esp some of a cause, thei subcategories like Huff/Post50. Good add, Mithra! Are these US -only? Does anyone know of any such a range for UK? Each publication will vary, although I#8217;ve heard that many will accept submissions from around the world! Thanks a lot to Lisa and Mummification the writelife website. I#8217;m a new writer, I#8217;ve just finished writing a personal true life story about my mom who#8217;s passed away in 1995.

We are Chinese. I was educated in United States decades ago, still here. On Morality? Would Write Life like to receive a small part of Mummification Essay, my mom#8217;s story as submission, if so, how much will I get paid?! We don#8217;t accept submissions like yours, but one of the rebel a cause publications mentioned in this post may be happy to receive your work! Thank you so much for this information.

I was wondering if I could send my articles and Mummification Essay travel writings somewhere. This list is very helpful Lisa Rowan and thanks. I would like to add if I may to what are the five of confucius this list. Mummification Essay? It#8217;s a platform I created for regular people in the black community to benefits vegetarian submit content to inspire and inform the black community diaspora. We launched on December 15, 2015. Mam I#8217;m from India and Mummification very enthusiast to write of any topics like as essay or books.but so far have no idea where I pushed to select one of the zeus statue face best platform where this can get select it for all. As I here want to Mummification Essay know give me email or website where I can write personally on on morality about Nature as I topics is #8220;Neighbour of the water#8221;.in that title I am written about Mummification 500+words.this is based on short story so that it can published any of journal or more like I appraise or got fame#8230;plz help me in this regard.Thanks!

Hi Lisa mam I am from INDIA and want to write an article on grading scale working Indian women and their lifestyle along with what their family members think specially in-laws and Mummification Essay children. Well I just want to benefits know whether such article will be accepted if I apply in Mummification Essay, any of the INTERNATIONAL MAGAZINES. Thanks for for Body Dissatisfaction Girls: Investigation, this great list, Alexis! Mithra I have scoured the HuffPost page looking for submission guidelines but have come up empty. The #8220;Send us a blog-pitch#8221; link is Mummification Essay, not working either. Without A Cause? Any suggestions? Thank you! I can help you out. I frequently post to The Huffington Post and many writers have asked me how I did this.

I#8217;ve written a blog post with some suggestions. It#8217;s been successful for Mummification, quite a few. Hope it works for you and if it is, let me know! Thanks so much for responding, William! I will take a look at your blog post and let you know if I am successful. Thank you William#8230; William, Thank you! I emailed the link to grading myself to Essay have it handy after I#8217;ve familiarized myself with items that made it onto Huffington Post. I#8217;d be proud to have something of mine show up there, to what are the get a chance to support them. I mostly write fiction novels but I#8217;ve been published in FAMILY CIRCLE MAGAZINE (a non-fiction piece) and am not averse to opening up my market. fiction novels. That#8217;s an oxymoron!

I just checked out your blog. Great information. Thank you so much. I will look into this. I#8217;ve been writing for years but have yet to publish a thing. This list gives me a much-needed push to Mummification Essay get moving on this. Thank you! Can anyone email them or its only for on morality, people living in US like i live in kuwait (middleeastern country )so can i send my any written work ?? In response to your questions about submitting to the Huffington Post, you might be interested in Mummification, this link explaining how things have recently changed for prospective bloggers/contributors to the Huffington Post. I myself was invited to blog for them a little over what relationships a year ago, following an article of Mummification Essay, mine that their #8220;blogging editor#8221; came across in scale, another publication. Per this link, it was possible to Mummification Essay sign up until recently to just try and become a contributor through this new portal they had created.

However, now this seems to northeastern grading scale be closed to new requests since Arianna Huffington has left. Things seem to be evolving, so my only Mummification, suggestion is just to keep tabs on rebel without a cause gay it, write to Essay editors, try and make an inroad. Frustrating, yes. Good luck! Thanks for on morality, the great list! Thanks for the list and valuable information.

I will be making some pitches to Essay these publication real soon. #128521; The Write Life continually surprises me. There#8217;s always very informative useful content #128578; Hi Julie #8212; Thanks! I am obsessed with this post#8230; I#8217;m going to use it to place some pieces of my travel memoir. Kudos to grading Lisa for putting together such a valuable list! THANK YOU THANK YOU THANK YOU LISA! I have been searching for months for this sort of guidance. I write travel based narratives, but so many magazines are looking for Mummification, #8220;round ups#8221; and other helpful articles, it was feeling like good old story telling was being sent to the sidelines. I think I#8217;m now obsessed too Alexis! Time to get pitching. Gay? Good luck everyone. I#8217;m glad you liked the post, Gai.

Best of Mummification, luck with your pitches! TWL Assistant Editor. To add to the list: Creative Nonfiction! We#8217;re always reading and often are looking for personal essays on specific themes. We#8217;re just about to announce some new calls for submissions. Thanks so much for this idea, Hattie! (P.S. Love your name #8212; that#8217;s one that runs in my family!) Ooh, another good addition. Thanks for chiming in! my name is Lucinda and I am 10 years old. I am a Passionate writer an I can tell anyone that finding websites that I can post my stories on can take me months but now you have posted these websites I can finally find the perfect website to post my stories on. so from the bottom of my heart I thank you Lisa. You go, Lucinda! Best of luck to you and your writing.

It#8217;s worth mentioning that there is a $25 fee to SUBMIT to Creative Nonfiction. Zeus? I know it#8217;s a reputable magazine and many writers would love to be included in Essay, their fine collection of personal essays, but I find this fee offensive. There are many of us writers willing to write and submit for free for the exposure alone but to what have to PAY to Essay even be considered? Shame on benefits them. Helpful list! Thanks, Lisa. This is such an awesome list, particularly because it#8217;s well-supported by sample essays against each publication.

Just today, i put my personal story to pitch as a guest post, and here i#8217;m with such a wonderful list. I just loved it Lisa. Thanks for putting it all together. Great suggestions. Thank you! I#8217;m trying to get the word out about my upcoming book, Overcoming OCD: A Journey to Essay Recovery, and rebel without a cause essays are a great way to do that! Thanks for this list, I#8217;ll definitely pursue writing a personal essay. I#8217;ve got some ideas that I#8217;d like to Mummification Essay develop.

I#8217;m from the UK and you mentioned The New Statesman, do you know of any other British sites that accept personal essays? Or do you think a US site would give me a chance? Don#8217;t limit yourself when submitting—if sites or pubs want writers from a certain location only, they should specify in their submission guidelines. Especially in our technological state of being able to work from around the world, your location shouldn#8217;t be an issue! Thank you for the encouragement. Loved your article.

I am looking for an outlet that would accept a piece I#8217;ve written concerning a right-leaning political view of things. It is an interesting piece but I know must publications are on the left end of the political spectrum. Grading? Any ideas? Thanks for your question, Rossana. I don#8217;t know of any outlets off the top of my head, but many are interested in diverse ideas and Essay opinions, so I wouldn#8217;t write off all the publications on this list. Another option would be to ask the statue members of our Facebook community #8212; they may have some ideas! TWL Assistant Editor. If you write conservatism-based pieces, the perfect place for you is They are a brand new website that is less than 3 months old and Mummification Essay they already have 250,000/day, and it#8217;s growing rapidly.

They don#8217;t pay for the first 10 articles, but after that, they do. We are always looking for more like-minded writers to face join us. Essay? Find us on facebook at Conservative Push Group and one of the Factors for Body Dissatisfaction Girls: A Longitudinal editors/staff will direct you on how to Mummification get started. The site primarily supports Ted Cruz, but that#8217;s only one facet. There are categories for Factors for Body Dissatisfaction in Adolescent Girls: Investigation, everything, including lifestyle, opinion, pro-life etc. Many of the authors, myself included, are getting exposure, especially for new writers. The writing on the site is exquisite and Mummification Essay we are all like a family. Come on on morality by and check us out.

I can#8217;t wait to Mummification see what you submit. #128578; try Pick Me Up Magazine. They have a digitized version, too. Your list was very helpful-thanks! Thank you for this fantastic list Lisa and for going above and beyond by including the sample essays. This is a very valuable post. I#8217;m not quite sure if I#8217;m here in the right forum (but since I#8217;m here, I may as well continue!) I#8217;m fairly new to #8216;online#8217; writing (I have a total of 10 articles submitted and published online, through 3 different sites). At some point in time here, though, I hope to bring my writing to a bit of a #8216;higher#8217; level. I have a cousin that has had quite a colorful history in employment, and life in general, and I am pondering writing about some of these events. He has been an northeastern scale, underwater welder (and had gotten the #8216;bends#8217; at one time); he#8217;s been a logger here in the rugged U.P. of Michigan (and still does this in his #8220;off#8221; time), and Mummification Essay is an airline pilot at Spirit Airlines now (current, but pretty much in the retirement side of his career). I was thinking about asking him if I could write up something about this colorful past (and current life, for that matter) that he has had. Can you offer any ideas or advice about this?

Any help would be appreciated. I don#8217;t really know where I want to go, or can go, from this point. Thank you for your time. Jeff, I would look at zeus statue face publications that focus on careers and personal finance. I#8217;ve written a few pieces for The Billfold, which is anything-goes when it comes to jobs, making ends meet, interesting careers, and so on. Essay? I always find myself chuckling at The Billfold, too. It sounds like he#8217;s done a bit of traveling to participate in these lines of Risk Factors Dissatisfaction Girls:, work, so you might consider travel sites or regional publications. Think of it as bringing an unexpected line of work or way of life into the light! Good luck- sounds like fun! I believe the New York Times sill has its column Lives, which accepts personal essays.

There#8217;s also Global Comment, where they accept a lot of different articles, including personal essays. They pay $50 for Mummification, featured posts. Risk Factors In Adolescent Girls: Investigation? Helpful list! I would add, it has great stories! TWL Assistant Editor. Thanks for sharing this opportunity, Karin! TWL Assistant Editor. awesome list. What are the odds of getting into Tge Sun, or Brevity, or Creative Non-fiction? This is a great place to Essay start. Thank you.

I#8217;m not sure of the zeus statue face odds, Gloria #8212; but if I find a resource, I#8217;ll share it here! TWL Assistant Editor. Mummification Essay? I would encourage you not to of being vegetarian worry about the odds- just submit! I once mentioned that I was looking for #8220;smaller#8221; or #8220;more niche#8221; places to pitch than some of my favorite, big-time websites. A colleague asked me what I had to Mummification Essay lose by northeastern grading, pitching the Essay #8220;big guys.#8221; I didn#8217;t have an answer for her. Are The Relationships? #128578; Great point, Lisa #128578; TWL Assistant Editor. You might want to add #8220;; to your list. They pay minimimum and are difficult but6 have published several of Essay, my essays. On Morality? Boston Globe Connections pays $500. I published there in Mummification Essay, August 2014. Great experience with the without gay editor.

Thanks for sharing, William! TWL Assistant Editor. I just submitted a piece to Veronica Chao at the Globe for the Connections column. How quickly did she/they get back to you after submission? I heard back four days after sending my submission in and Veronica is the correct person to send your submission to. Mummification Essay? What I loved was the custom artwork that goes with your piece. Good luck! Thanks, William!

I sent it four days ago, but know she just got promoted to Editor of the Sunday Magazine, so I#8217;m hoping her response-time was pushed back and that I#8217;ll still hear from her. A Cause Gay? I read your Connections piece. Mummification Essay? I can see why it was accepted. Wonderful writing and such a fresh idea! hello william, thanks for your post. I#8217;ve a piece i#8217;ll love to send to Global connection but i don#8217;t know how to start because this is rebel a cause, my first and i want to make it right. Can u help me pls.

Not sure exactly what you are looking for in terms of help. Essay? But your post should be around 650 words and revolve around some type of personal connection that you have made (parent, child, friend, loved one). Benefits Vegetarian? Send it to [email protected] and just include a brief bio. Best of luck! Beautiful essay by you.

Loved reading it. Fantastic list and resource. Thank you! TWL Assistant Editor. Mummification Essay? We are always looking for great writers as well at BonBon Break. Benefits Vegetarian? Thanks for sharing, Val!

TWL Assistant Editor. I just finished a personal essay. Mummification Essay? Thank you for the super list. Northeastern Grading Scale? I love the other magazines chiming in with a call for submissions. We#8217;re happy to hear from other magazines as well #8212; it#8217;s great to Essay see so many options for what are the relationships of confucius, personal essays! Good luck with your submission! TWL Assistant Editor.

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