Involvement of the xdh gene in Gene Polymorphisms Involved in Manifestation of Leucopenia, Digestive Intolerance, and Pancreatitis in Azathioprine-Treated Patients

An estimated 10-28% of individuals treated with thiopurines get a severe drug reaction from them. Myelosuppresion is one of the severe reaction that patients experience (Myelosupression is also termed leucopenia). The main objective of this research was to study the variant alleles of thiopurine S-methyltransferase, inosine triphosphate diphosphatase, and xanthine dehydrogenase (xdh) and their relationships to adverse drug reactions to azathioprine therapy. 188 Caucasian patients with inflammatory bowel disease were subject to a standard dose of azathiprine. Statistical results were obtain by the use of standard PCR and Quantitative PCR and with the use of Fisher’s Test and the odds ratio calculation. In regards to our xdh gene, more results are being waited on but it is believed that xdh gene plays a role in the development of pancreatitis (inflammation of the pancreas).

Works Cited

Source: http://www.ncbi.nlm.nih.gov/pubmed/22535280

Wroblova K, Kolorz M, Batovsky M, Zboril V, Suchankova J, Bartos M, Ulicny B, Pav I, Bartosova L.

Department of Human Pharmacology and Toxicology, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno, Palackeho 1/3, 612 42, Brno, Czech Republic.

Preparation of successfully amplified xdh regions from DNAs 4-34 to 4-44

Purpose

Experiment performed by Mahmud Abdallah and Sasha LeSure.  This experiment required amplification of the xdh region of DNA with the Polymerase Chain Reaction with the use of specific primers. Then we ran our samples using Gel Electrophoresis and the results are shown in the picture below. We want to obtain the sequence of our extracted DNA afterward.

Notes: We used Ethidium Bromide to obtain a sharper image of our results because our initial stain was faint. The products desired are on those next to the highest most DNA marker.

Samples 1-8 are approximately 1000 base pairs compared to our Gel Marker (GM).

Samples 9,10, and 11 did not produce any efficient products so we did not count them in our calculations (possibly due to a low quantity of DNA).

On the bottom of the marker we have PCR artifacts possibly from primers priming off each other.

Figure 1: Gel Electrophoresis of DNA samples 4-34 to 4-44. H2O was used as a negative control indicating no contamination. Samples 4-42 to 4-44 produced insufficient results. Organisms DNA indicated in figure 2.

Lane Number	DNA Code	Genus Species	PCR Results
GM	–	Gel Marker
H2O	–	Negative Control
1	4-34	Polystachya longiscapa	++
2	4-35	Cyrtorchis monteirae	++
3	4-36	Tridactyle tridentata	++
4	4-37	Jumellea comorensis	++
5	4-38	Aerangis biloba	++
6	4-39	Phalaenopsis stuartiana	++
7	4-40	Phalaenopsis amboinensis	++
8	4-41	Phalaenopsis gigantea	++
9	4-42	Paraphalaenopsis serpentelingua	–
10	4-43	Adenoncos sp.	–
11	4-44	Papilionanthe pedunculata	–

Figure 2: Gel Electrophoresis results including organisms the DNA was extracted from, the code for each DNA sample, and the results of the PCR run.

Gastrochilus acutifolius

Photo taken by Mahmud S. Abdallah of specimen in research collection.

Gastrochilus acutifolius (Lindl.) Kuntze. belongs to the tribe Vandeae and the sub tribe Aeridinae. This plant can be found in Assam India, the Himalayas, Nepal, Myanmar and Vietnam growing at an elevation between 700-900 meters above sea level. This beautiful orchid’s leaves can reach lengths up to 2 inches long and possesses 8 flowers with each having a length of approximately 2 cm with a very distinct lip not seen in my other orchids. G. acutifolius is an epiphytic ¹  species which can grow in warm as well as hotter environments. It requires prompt watering of the roots and exposure to plenty of sunlight. It is noted, however, that it should not be watered after the flowers have bloomed. It lives in areas surrounded by other plants such as coniferous ²  forests and plenty of mosses. G. acutifolius usually starts blooming its flowers in fall and it’s flowers are described as fragrant.

Source: http://www.orchidspecies.com/gastacutifolium.htm

Vocabulary:

¹Epiphytic: a plant that grows upon another plant (such as a tree) non-parasitically or sometimes upon some other object (such as a building or a telegraph wire), derives its moisture and nutrients from the air and rain and sometimes from debris accumulating around it, and is found in the temperate zone (as many mosses, liverworts, lichens and algae) and in the tropics.

²Coniferous: One of 13 or 14 division level taxa within the Kingdom Plantae. They are gymnosperms. They are cone-bearing seed plants with vascular tissue; all extant conifers are woody plants, the great majority being trees with just a few being shrubs.

____________________________________________________________________________________________________

References:

http://orchids.wikia.com/wiki/Gastrochilus_acutifolius (Orchids Wiki)

http://www.orchidspecies.com/gastacutifolium.htm (Orchid Species)