The Blue Mexican: A Novel
However, the effectiveness of breast cancer detection can be improved by including tests that identify mutations in the BRCA genes Therefore, it would be desirable to have alternative low-cost options for the detection of mutations.
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This method was capable of detecting four positive BRCA1 exdel mutation carriers, which were previously analyzed by our end-point PCR method. This was the expected result, since the probe was designed to be complementary to the mutant allele, specifically in the recombination region between introns 8 and 12, which is a unique sequence in the human DNA. High prices are a limitation for those who wish to know their genetic status regarding BRCA1 exdel mutation. Using qPCR allows high sensitivity since the reaction can be carried out only with 2.
The fact that a small amount of DNA is required for the assay gives an extra strength to this method, since DNA can be extracted from small biological samples taken in a non-invasive manner, such as urine, epithelial cells from the mouth or hair roots. Peripheral blood guarantees a high DNA yield, but its extraction is painful and uncomfortable for the patient, especially when the quality of the veins is not good due to prior chemotherapy. It is a fast method since in 2 h a total of 90 samples can be analyzed in a single experiment, including the interpretation.
Biallelic mutations in both BRCA genes are rare 32 , Therefore, deleterious mutations in these genes are mainly heterozygous However, biallelic pathogenic mutations have been reported in individuals with ovarian cancer. Nevertheless, those patients are compound heterozygous, this is, they carry mutations in both BRCA1 copies, but in different regions for example: c. Biallelic mutations in BRCA genes could increase the risk of cancer, as well as leading to more severe phenotypes 32 , To date, no individual has been found homozygous for the BRCA1 exdel mutation.
Most probably the latter would not be compatible with life since great quantity of genetic material is lost. However, it is important to detect any region of the wild-type allele at the same time that we detect the mutant allele. This is also helpful as a control of DNA presence in the sample. Nonetheless, in our study, we performed the PCR of the variant allele and the PCR of the wild-type allele in separate reaction wells.
So, a sample will be defined as heterozygous only if it appears in both clusters in the AD plot Fig. Consequently, as future study, we aim to standardize a multiplex qPCR capable of detecting the variant allele recombination region and the wild-type allele a region from exons 9 to 12 in one single reaction, saving time and reagents.
Emotions and Otherness in D. H. Lawrence’s Mexican Fiction
However, if a different recombination region is discovered, or if any additional mutations arise in this region, our method would have to be modified accordingly and readapted to the newly reported genetic variants. Compared to the reported methods used to detect the BRCA1 exdel mutation, our propose represents a suitable option for testing given the advantages already mentioned.
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The software of the qPCR thermocycler analyzes the results at the end of the run, displaying the presence or absence of the mutation and therefore, making the interpretation an easy step. Moreover, working with qPCR avoids the risk of post-amplification contamination, since only one process is needed. On the other hand, end-point PCR requires a first step of amplification, followed by the run of the samples on an agarose gel, a process in which the samples can be cross-contaminated Additionally, it does not require further manipulation of the sample, for example purification of the amplified product, sequencing-PCR and capillary electrophoresis.
The analysis of results is faster and the identification of the heterozygous carriers is an easier task Another method used to detect deletions is MLPA, which has the great advantage of detecting more than one mutation since it analyzes different regions of the genome simultaneously. However, for the detection of the BRCA1 exdel mutation, this method has the disadvantage of requiring a large amount of DNA input — ng , increased sample handling, carrying an additional risk of cross-contamination.
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The method we propose will allow the determination of a high number of samples at the same time, which will be a great advantage in future studies of association or prevalence. Additionally, it is a safe, specific and more affordable option for those wishing to be tested for the mutation. The present study was supported by the University of Monterrey grant no. RBRLC conceived the study, directed, planned and analyzed the data and wrote manuscript. National Center for Biotechnology Information , U. Mol Med Rep. Published online Jun 6.
Author information Article notes Copyright and License information Disclaimer. Received Dec 5; Accepted Apr This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
Abstract In , according to the National Institute of Statistics and Geography INEGI , malignant breast tumors were the first cause of cancer fatality in women 6, fatalities in Mexico, whereas 2, fatalities in women were due to ovarian cancer. Introduction Current diagnostic tools for detection of breast cancer have contributed to reduce its mortality. Open in a separate window. Figure 1. Figure 2. Figure 3. Discussion Currently, different methods are used for the early detection of cancer, such as analysis of circulating tumor cells CTCs , as well as RNA exosomes 27 , proteins 3 and genomic profiles of circulating cell-free tumor DNA ctDNA from liquid biopsy 27 , Table I.
Conventional PCR thermocycler and capillary sequencer. Amount of DNA required per reaction 2. Funding The present study was supported by the University of Monterrey grant no. Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. References 1. Query results: Basic tabulations. Identification and validation of salivary proteomic signatures for non-invasive detection of ovarian cancer.
Two cups cooked chopped beef, one-half cup green chile pulp, one-half cup suet, tablespoon lard, one cup tomatoes, one button garlic, one-half cup chopped onion. Cook until thick, serve with frijoles. One-half cup chopped salt pork, one-half cup suet, add one-half cup chopped onion, one teaspoon aregano, one-half teaspoon safron, fry all together; add two cups cooked ground beef, two cups frijoles, two cups red chile pulp, one cup water or three cups water and four tablespoons chile powder, salt to taste and cook forty minutes.
Boil a fat chicken in small amount water with garlic button, or slice of onion to flavor. When tender chop the meat in small pieces, heat some of the chicken fat skimmed from the liquor and cut from chicken, add flour, bit of onion, add one cup green chile pulp, two cups of water chicken was boiled in, salt; add chopped chicken, stew few minutes, serve on Spanish rice. Cook beef, veal or chicken with parsley, carrots, celery, onions, bay leaf.
Let it cook very slowly until tender, then cut meat into small pieces, strain the juice, add to every pint one-half cup chile pulp and one cup of cooked and mashed frijoles, add tablespoon butter or lard, then meat, and if too hot add cup tomatoes, cook few minutes longer. Chop one pound of beef, pork or chicken, add a little chopped tallow or one tablespoonful of lard and a little salt; fry in a pan until tender; chop again very fine; return to pan; add a little warm water and pulp of two red chiles; stir and fry few minutes.
Add to one quart of cornmeal two tablespoonfuls of salt, two tablespoonfuls of lard, and boiling water to make a thick dough. Cut off about one inch of corn husk stalk ends and soak in hot water ten minutes; dry and rub over with hot lard. Put a layer of dough on the husk about four inches long, one and one-half inches wide and one-fourth inch thick; along the center spread two teaspoonfuls of the prepared meat; roll and fold the small end of the husk; place them folded end down in a strainer over hot water.
Cover and steam several hours. Soak some trimmed corn husk bought in Mexican stores for several hours in cold water, then boil until soft, remove; dry on cloth, and rub with lard. Cut up a fat chicken, cook until very tender in just enough water to leave about four cups. Chop up cooked chicken, add corn meal or masa to boiling hot chicken broth until a thick dough; add salt to taste, one tablespoon chile powder, or chile sauce No. Now to all the chicken add enough chile sauce No. In center of one larger husk place a large kitchen spoonful of chicken; spread over this one tablespoonful of dough; place another husk spread with dough; continue placing husk around on all sides until about ten are used.
Contested Indigeneity in the Mexican Colorado Delta
Tie ends together with a strip of husk and place on end in a colander over boiling water for two or three hours, or place some corn husk in bottom of vessel, pile tamales on top, pour in about a quart of water, bring to a boil and steam slowly for three or four hours. Make a crust of two cups of hot water, one-half cup lard, one teaspoon salt, and cornmeal to make thick dough.
Spread on bottom and sides of pie tin. Mix beef, pork, veal or chicken with plenty of suet for shortening—any two of above meats will do—measure two cups of meat, add two tablespoons of cornmeal, one-half cup of chile pulp, one teaspoon salt, one-half cup meat stock or water. Cook for a few minutes, fill pie crust, spread more dough over top. Make little balls the size of marbles set around edge. Pour a little melted lard over the top, sprinkle with chile powder; bake about thirty minutes.
Ground beef fried in lard and suet, add chile sauce No. Prepare cornmeal or masa by pouring on boiling hot water, salt to taste, add one cup of lard to four cups meal or masa, add few olives and raisins if desired.
Prepare as for chicken tamale recipe. Texas tamales made as above with olives and raisins left out and spoonful put in one long shuck, fold ends over and press flat. There are firms in California putting up a specially prepared corn meal for tortillas.
There are chile powders and canned red and green hot chile peppers; also the red sweet pepper, called pimiento, which makes it possible to cook Spanish dishes anywhere. A complete lesson—Fry a strip of bacon for each diner; remove from fat and keep hot. Cook until tender, in the fat, one tablespoon minced onion and sliver of garlic button young green onion is best , one tablespoon minced green sweet pepper, same of canned sweet red pepper pimiento , one tablespoon parsley, four sliced ripe olives, four seedless raisins, six mushrooms.
Add when done, level teaspoon flour, then add one cup tomato—raw or canned; cook few minutes, salt to taste, add one teaspoon chile powder or two tablespoons red chile sauce. Use the above amount for every two eggs. Keep hot and prepare eggs by breaking in separate bowl whites and yolk; beat yolk first very stiff till light in color, add tablespoon water to each yolk and one-eighth teaspoon salt. Beat whites stiff, add small pinch cream tartar while whipping. Fold three-fourths of the whites into the yolks—do not break up too fine.
When nearly done, set under gas broiler or in very hot oven for a few minutes until seared. Remove, and with spoon, put prepared filling on one-half of omelet, fold other half over, turn out on a hot platter. Put rest of whipped whites on top to form a circular crown around edge. Sprinkle with salt and bits of butter; set back in oven and brown but not too deeply.