Abstract
Acid soluble proteins from 23 samples of normal human gastrointestinal mucosa derived from four normal adult organ donors were extracted and subjected to specific radiommunoassays for transforming growth factor alpha (TGF alpha) and urogastrone epidermal growth factor (URO-EGF). All tissues were found to contain immunoreactive TGF alpha and levels ranged from 57 to 4,776 pg-1 wet weight of tissue. Although levels varied between tissue donors, the distribution of TGF alpha throughout the gastrointestinal tract appeared similar in all cases. URO-EGF levels were much lower (0-216 pg g-1 wet weight). TGF alpha levels in extracts of gastrointestinal mucosa from a 7-year-old female donor were higher and the observed distribution was markedly different from adult levels. URO-EGF was not detected in mucosal or submucosal tissue extracts from this patient. Further studies in juveniles are indicated.
Highlights
Tissues were largely derived from only three adults, and absolute levels varied between individuals, the distribution of hTGFa throughout the gastrointestinal tract mucosa appeared similar. hTGFa levels declined significantly from the gastric mucosa to the duodenal mucosa (396pgg-') and gradually rose again through the ileum (1,290 pg g') and ascending colon (2,173 pg g-') to decrease again through to the sigmoid colon mucosa (530pgg-')
There was no apparent pattern in the distribution of hURO-EGF along the gastrointestinal tract mucosa
To verify the nature of the observed immunoreactivity, reverse phase chromatography was carried out on samples of gastric mucosal extracts (Figure 2). hTGFa and hURO-EGF immunoreactivity were found to coelute exactly with hTGFa and hURO-EGF standards previously applied to the column. hTGFo eluted at 25% acetonitrile and hURO-EGF at 30% acetonitrile
Summary
TissuesWith the permission of HM Coroner and the Coroner's pathologist for North Staffordshire, 30 samples of gastrointestinal tissues of lengths 15-30 cm were obtained from five normal organ donors (three male and one female adults (age range 19-44 years) and one 7-year-old female) as quickly as possible following organ donation and in all cases within 90 min of cessation of cardiac action. Tissues were homogenised using a Silverson Laboratory Mixer (Silverson Machines Ltd, Chesham, Bucks., UK) in 4 ml g-' of a solution comprising 375 ml of 95% (v/v) ethanol, 7.5 ml of concentrated HCI, 33 mg of phenylmethylsulphonyl fluoride and 1.9 mg of pepstatin. The volume was adjusted to 6 ml g-' with distilled water and extracted overnight at 4°C. Mixtures were centrifuged and pellets reextracted overnight with 4 ml g-' (original weight) of a solution containing 375 ml of 95% ethanol, 7.5 ml concentrated HCI and 105 ml distilled water. 1 paper and redissolved in 1M acetic acid (3.5 ml-' of original tissue weight). Extracts were dialysed extensively against 0.1 M acetic acid (Spectrapor tubing, molecular weight cut-off 3,500, Spectrum Medical Industries, Los Angeles, CA)
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