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Biomedical subjects

R Seth

Publications and source records attributed to R Seth.

At least 19 recordsLinked to original sources
PubMed2006

A biological process that reduces metals in municipal sludge to yield sulphur enhanced biosolids.

Heavy metal contamination can often restrict or prohibit land application of municipal sludge. In the present investigation a continuous biological process with powdered elemental sulphur as energy source was examined for solubilization of metals from anaerobically digested municipal sludge. The results showed that the continuous process can be designed to achieve both metal solubilization and efficient sulphur utilization. With a sulphur addition of 1.5 g l(-1) and an HRT of 14 days, solubilization efficiencies of 50, 33, 48, and 74% were obtained for cadmium, copper, nickel, and zinc, and about 80% of the added sulphur was oxidized. Addition of ferrous sulphate as an additional energy source did not improve the performance of the process. The residual sulphur in the metal decontaminated biosolids was in balance with the phosphorus content, potentially making it an effective sulphur fertilizer for soils deficient in the nutrient.

Biodegradation, Environmental↗
PubMed2005

Human papillomavirus infection in women who develop high-grade cervical intraepithelial neoplasia or cervical cancer: a case-control study in the UK.

Human papillomavirus (HPV) testing might identify older women who could be withdrawn from the cervical screening programme, or require less frequent screening. A case-control study using the United Kingdom cervical screening population was set up to help address this issue. Cases comprised 575 women who developed cervical intraepithelial neoplasia (CIN) grade 2 or worse over a 13-year period following a cytologically normal baseline smear, and were stratified by age group ('under 20', '20-39' and 40 years or over). Controls (n=601) were women who remained disease free over this interval and were the same age on average as cases. DNA was extracted from the baseline smears and tested for HPV by PCR using GP5+/6+ consensus primers. HPV+ samples were tested for HPV types 16 and 18 using specific PCR primers. In all, 27.0% of cases tested positive for HPV at baseline, compared with 15.4% of controls (odds ratio (OR)=2.00; 95% confidence interval (CI), 1.50-2.68). Among women aged 40 years or over, the OR for HPV 16 was 8.95 (95% CI, 2.63-30.4). These results support the need for further cervical screening of HPV- older women, as many of the cases were HPV- at baseline.

Adult↗
PubMed2004

In vitro assessment of copper-induced toxicity in the human hepatoma line, Hep G2.

Copper, though essential, is highly toxic when present in excess, as in Wilson disease, a genetic disorder of hepatic copper metabolism. We hypothesized that mitochondria are a major target of copper-induced cytotoxicity in Wilson disease. We used the human hepatoma line Hep G2 to examine copper-mediated cytotoxicity and three different methods to assess organelle damage: MTT assay (mitochondria), neutral red (NR; lysosomes) and Trypan blue exclusion assay (TB; plasma membrane). For all assays, cells at approximately 60% confluence in microtitre plates were incubated with CuCl(2) (concentration range: 50-100-150-200 microM) for 24 or 48 h. Results were expressed as percent of untreated control. At 24 h, cytotoxicity as detected by NR assay was significantly higher at all concentrations of copper than for MTT or TB ( p<0.005 at all concentrations). Cytotoxicity as detected by MTT was higher than that detected by TB at all concentrations except at 200 microM (p<0.05 for 50 microM, p<0.005 for 100 microM, p = 0.001 for 150 microM). Results at 48 h were similar (NR versus others: p <0.001 MTT versus TB: NS except at 150 microM where p<0.01). We investigated reactive oxygen species (ROS) production in copper-associated hepatocytoxicity by incubating sub-confluent cells with 2('),7(')-dichlorodihydrofluorescein diacetate dye plus copper (concentration range: 0-200 microM) for 1-1.5 h. Copper, but not zinc, produced significant increases in ROS (p<0.001). In summary, Hep G2 lysosomes appeared more susceptible to Cu-mediated damage than mitochondria; the cell membrane was highly resistant to damage.

Carcinoma, Hepatocellular↗
PubMed2004

Changes in expression of AVT and AVT receptor (VT1) gene in hypothalamus and shell gland in relation to egg laying in white leghorn hen.

Oviposition is a complex phenomenon involving various regulatory mechanisms at the neuroendocrine levels. Present study was designed to access the changes in arginine vasotocin (AVT) and its receptor (VT1) gene expression in relation to the time of egg laying of white leghorn hen. The expression of AVT gene (Northern blot analysis and in situ hybridization) in the hypothalamus and localization of ir-AVT in the magnocellular neurons of paraventricular nuclei was studied 2 h before (-2 h), immediately after (0 h) and 2 h after (+2 h) egg laying. Simultaneous changes in the AVT and VT1 receptor gene in the shell gland, which finally responds to AVT for smooth muscle contraction and expulsion of egg, were also determined by semi-quantitative reverse transcriptase-polymerase chain reaction. The findings indicated increased hypothalamic AVT gene expression immediately after egg laying (0 h) when compared to 2 h before and 2 h after egg laying. AVT receptor gene expression in the shell gland also followed the same pattern. However, AVT gene expression in the shell gland, unlike that of hypothalamus was higher at -2 h compared to 0 and +2 h of oviposition. While highly significant increase was noted in plasma AVT concentration at the time of egg laying, other parameters such as plasma osmolality and ionic concentration (Na(+), K(+), Ca(2+), and Cl(-)) did not show any change. It is suggested that in addition to increased hypothalamic AVT transcript and peripheral release, local synthesis of AVT in the shell gland (paracrine release) may contribute to the contraction of shell gland smooth muscles during egg laying. Moreover, these findings clearly indicate temporal correlation of AVT and its receptor gene expression in different tissues during oviposition.

Animals↗
PubMed2004

Expression of hypothalamic arginine vasotocin gene in response to water deprivation and sex steroid administration in female Japanese quail.

Arginine vasotocin (AVT) is a neurohypophyseal hormone involved in reproductive function and control of osmoregulation in birds. In view of the dual function of AVT, the present experiment was designed to observe the effect of water deprivation (WD) and sex steroid [estradiol benzoate (EB) and testosterone propionate (TP)] treatment independently, as well as simultaneously, on the profile/activity of the hypothalamic AVT system. WD resulted in a significant increase in plasma osmolality, sodium ion concentration and AVT concentration, but administration of sex steroids had no significant influence on these parameters. By contrast, the amount of hypothalamic AVT transcript (northern analysis) and the size of immunoreactive vasotocin (ir-AVT) neurons and hybridization signals (in the form of silver grains), representing AVT mRNA in corresponding neurons of paraventricular nuclei (PVN), increased significantly in all the treated groups compared with controls. Our findings indicate that although sex steroid administration has no effect on plasma osmolality and AVT concentration, unlike water deprivation, it may stimulate the profile/activity of AVT neurons of PVN, supporting the possibility of sex steroid receptors on these neurons. It is concluded that in quail, osmotic stress not only upregulates the expression of the AVT gene in existing neurons but also recruits many more neurons to increase the rate of AVT synthesis and secretion, while sex steroids appear to have a stimulatory effect only on the existing number of neurons and only at the level of transcription/translation and hence may influence/modulate hypothalamic AVT gene expression in response to osmotic stress. This study also suggests an interrelationship between reproduction and AVT system/function in birds.

Analysis of Variance↗
PubMed2002

Selective reduction of intestinal trefoil factor in untreated coeliac disease.

The trefoil factor family (TFF) encompasses small peptides of which intestinal trefoil factor (ITF) is expressed specifically in goblet cells of the small and large intestine. Previous studies have shown that ITF plays an important role in mucosal protection and repair. Coeliac disease represents a model of immune-mediated small intestinal inflammation and damage, with recovery on gluten-free diet. The aim of this study was to investigate the expression of ITF in the distal duodenal mucosa of subjects with coeliac disease, before and after treatment with a gluten-free diet. Expression of ITF and mucin in the distal duodenal biopsies from treated (n = 11) and untreated (n = 9) coeliac subjects and controls (n = 8) was investigated by immunohistochemistry and semiquantitative PCR. In untreated coeliac disease, there was reduction of ITF immunoreactivity in goblet cells but mucin expression was preserved. Mucosal recovery on gluten-free diet was associated with increased ITF immunoreactivity in goblet cells. There was also reduction in the expression of ITF transcripts, relative to MUC2 mRNA, in untreated coeliac duodenal samples, with recovery on gluten-free diet. Our study suggests that there is a selective reduction in the expression of the ITF gene in untreated coeliac disease. Recovery of ITF expression on a gluten-free diet suggests that the mucosal immune system regulates goblet cell differentiation and ITF expression in the human intestinal mucosa.

Adult↗
PubMed2001

Quantitative analysis of IL-10 and IFN-gamma mRNA levels in normal cervix and human papillomavirus type 16 associated cervical precancer.

Human papillomavirus type 16 is a major factor in cervical carcinogenesis. Inappropriate cytokine synthesis may direct the local immune response away from a type-1 (cellular) pattern and may subsequently contribute to the development and progression of precancer. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) using a competitive mimic was carried out to determine type-1 (interferon gamma (IFN-gamma)) and type-2 (interleukin-10 (IL-10)) cytokine mRNA levels in whole cervical specimens (without microdissection) from seven normal and nine HPV-16 positive CIN formalin-fixed paraffin-embedded tissues. Microdissection was used to measure separately the epithelial and sub-epithelial levels of IFN-gamma and IL-10 mRNAs in 11 specimens of normal cervix and 25 HPV-16 positive CIN (nine CIN 1, seven CIN 2 and nine CIN 3). IFN-gamma mRNA was lower in CIN than normal (p=0.04). IL-10 mRNA level in CIN was significantly higher (p=0.005) than in normal cervix (before microdissection). Epithelial IFN-gamma mRNA showed a significant decrease in all grades of CIN (median=3.58) compared with normal (7.74) (p<0.05), but there was no significant difference between the grades. A significant decrease in sub-epithelial IFN-gamma mRNA was found in CIN 1(9.81), CIN 2 (3.82) and CIN 3 (4.62) compared with normal cervix (27.35) (p<0.05). Also, sub-epithelial IFN-gamma mRNA was significantly lower in CIN 2 and CIN 3 than in CIN 1 (p=0.005 and 0.0005, respectively). IL-10 was detected in the epithelium of only one of 11 normal and one of 25 CIN, but sub-epithelial IL-10 was significantly higher in CIN 2 (0.08) and CIN 3 (0.26) than in normal (0.00) (p=0.036 and 0.0032, respectively). There was no significant difference in the sub-epithelial level of IL-10 between normal and CIN 1 (0.00) (p=0.96). Our results suggest that reduced epithelial and sub-epithelial IFN-gamma, as well as increased sub-epithelial IL-10 synthesis may play a role in the development and progression of HPV-16 associated cervical precancer.

Cervix Uteri↗
PubMed2001

Isolated human papillomavirus 18-positive extragenital bowenoid papulosis and idiopathic CD4+ lymphocytopenia.

We report a case of isolated extragenital bowenoid papulosis (BP) in a young man with an idiopathic low CD4 count. The lesions occurred on the dorsal aspect of his left middle finger and were not associated with genital involvement. Polymerase chain reaction studies of a biopsy demonstrated human papillomavirus 18. As far as we are aware, this is the first documented case of BP (genital or extragenital) associated with idiopathic CD4 lymphocytopenia.

Adult↗
PubMed2001

Reduction of indicator organisms during biological solubilization of metals: a pilot study.

Laboratory and pilot scale studies on the biological solubilization of metals from undigested raw sludge were carried out using elemental sulphur as the energy source. Metals (Cu, Zn, Ni and Cd), pH, sulfate and indicator bacteria [total coliforms (TC) faecal coliforms (FC) and faecal streptococci (FS)] were used to evaluate the system effectiveness. This paper does not report the detailed design and operational data of the pilot plant (available elsewhere [1]) but focuses on the biological tests carried out at the Main Wastewater Treatment Plant in Toronto from July to December, 1997. Since the indicator bacteria are injured from exposure to low pH and potentially toxic metal concentrations, improved alternative methods for their enumeration and their ability to repair acid-metal induced injury in a resuscitation medium (CASO tryptone soya broth) were also investigated. Resuscitation was not effective in repairing cells injured during bacterial leaching. The MPN technique (using lauryl tryptose broth) for enumerating indicator bacteria (total coliforms) gave higher counts and was therefore superior to the spread plate technique (using m-Endo agar) in recovering bacteria from acidic leached sludge (biosolids). However, the coliform bacterial counts from raw sludge were similar by the two methods. This study indicated that the biological solubilization process could significantly reduce the pathogenic indicators. Concentrations of TC, FC and FS in the leached biosolids from the solubilization tank were lower than concentrations in the raw sludge by 4 to 6 orders of magnitude and the finished product (biosolids) met US. EPA requirements for pathogen and metal concentrations for Class A biosolids to be used on agricultural land.

Agriculture↗
PubMed2000

Differential levels of granzyme B, regulatory cytokines, and apoptosis in Crohn's disease and ulcerative colitis at first presentation.

The mechanisms of tissue damage in ulcerative colitis and Crohn's disease may reflect disordered humoral or cell-mediated effector mechanisms, respectively. Mucosal biopsies from untreated inflammatory bowel disease patients and normal controls were analysed for the expression of granzyme B, a cytotoxic effector molecule specifically associated with cell-mediated immunity, and for regulatory cytokines. Messenger RNA (mRNA) was analysed by reverse transcription-polymerase chain reaction and enzyme-linked oligonucleotide chemiluminescence assay. Mucosal biopsies were analysed by immunohistochemistry for granzyme B protein and lymphocyte markers and for the presence of apoptotic cells by terminal deoxynucleotidyl transferase end labelling. Granzyme B mRNA was elevated in Crohn's disease, but not in ulcerative colitis or control mucosal biopsies. Granzyme B mRNA levels correlated with interferon gamma mRNA levels in Crohn's disease. Granzyme B was expressed in CD3+, CD8+ T cells in the lamina propria of Crohn's disease mucosa and there were significantly more apoptotic cells in the lamina propria in Crohn's disease. In conclusion, granzyme B-expressing T lymphocytes are present in the focal mucosal lesions of Crohn's disease, together with spatially related apoptotic cell death. These results support the hypothesis that T-cell-mediated cytotoxic effector mechanisms may play a role in Crohn's disease.

Adolescent↗
PubMed2000

Decreased synthesis and expression of TGF-beta1, beta2, and beta3 in epithelium of HPV 16-positive cervical precancer: a study by microdissection, quantitative RT-PCR, and immunocytochemistry.

Cervical carcinogenesis is a multistep process initiated by 'high-risk' human papillomaviruses (HR-HPVs), most commonly HPV 16. Transforming growth factor-beta (TGF-beta) inhibits epithelial proliferation and down-regulates transcription of E6/E7 genes of HPV. Altered TGF-beta expression may be important in carcinogenesis. Quantitative RT-PCR was used to investigate TGF-beta1, beta2, and beta3 mRNA levels in nine specimens of normal cervix and 15 cervical precancers (eight HPV-positive, including five HPV 16-positive). Immunocytochemical expression of TGF-beta1, beta2, and beta3 was examined in cervical intraepithelial neoplasia (CIN) positive for HPV 16 (26), and in HPV-negative, normal ectocervical epithelium (9); reserve cell hyperplasia (12); and immature (7) and mature (15) squamous metaplasia. The intensity of staining for TGF-beta1 was measured using grey-scale image analysis. Microdissection was used to investigate epithelial and stromal (excluding crypts) levels of TGF-beta1 mRNA in HPV 16-positive cervical precancer. Normal cervix, including reserve cells and immature and mature metaplasia, showed strong immunocytochemical expression of all TGF-beta isoforms. Expression was decreased in the basal third of the epithelium in CIN 1, in the basal and middle thirds in CIN 2, and in all layers in CIN 3. Quantitative analysis of TGF-beta1 expression showed that the changes in CIN compared with normal ectocervix and mature metaplasia were statistically highly significant (p<0.001, ANOVA). TGF-beta1, beta2, and beta3 mRNA levels showed a significant decrease only in the five HPV 16-positive CIN samples when compared with normal (p=0. 0034, 0.0033, and 0.029, respectively). TGF-beta mRNA levels in HPV 16-positive epithelium also decreased from normal through low-grade to high-grade precancer. Stromal TGF-beta1 was absent or very low compared with epithelial production and was not altered in HPV 16 precancer. Progressive loss of epithelial TGF-beta expression and synthesis may be important in HPV 16-associated human cervical carcinogenesis.

Epithelium↗
PubMed2000

Simulating the response of metal contaminated lakes to reductions in atmospheric loading using a modified QWASI model.

The changes in metal concentration following significant reductions in atmospheric metal loading of two nickel and copper contaminated lakes in Coniston Valley of the Sudbury Basin of Ontario, Canada were simulated by using steady-state and dynamic versions of a modified Quantitative Water Air Sediment Interaction (QWASI) Model. Metal partitioning and precipitation processes were quantified with the aid of US EPA's MINTEQA2 Model. The dynamic model successfully described the recovery of the two lakes and identified key input, loss and partitioning processes. A useful modelling strategy is to develop one or more steady-state models that give an approximate representation of conditions at defined times, then extend this to a dynamic version which can take into account the differing rates of response of components of the system. This modelling strategy can be used for designing and assessing remediation programs for metal contaminated lakes and watersheds.

Air Pollutants↗
PubMed2000

Bacteroides fragilis enterotoxin induces the expression of IL-8 and transforming growth factor-beta (TGF-beta) by human colonic epithelial cells.

Bacteroides fragilis toxin (BFT) has been shown to be capable of inducing intestinal mucosal inflammation in animals. Such inflammation may be responsible for diarrhoea, which occurs in some, but not all human carriers of enterotoxigenic strains of B. fragilis (ETBF). We have studied responses to BFT by different human intestinal epithelial cell lines and subsequently investigated the expression of IL-8 and TGF-beta by T84 cells. The latter were selected because their responses to BFT, characterized by morphological changes and cell death by apoptosis, were similar to those we have recently observed in primary human colonocytes. We show that BFT dose-dependently increased the expression of transcripts and protein of the polymorphonuclear cell chemoattractant IL-8. BFT also dose-dependently induced the release of TGF-beta, which has been shown to enhance the repair of the injured intestinal epithelium. However, the secreted TGF-beta was almost exclusively in the biologically inactive form, as determined by Mv1Lu bioassay. Our studies therefore suggest that exposure of colonic epithelial cells in vivo to high concentrations of BFT can initiate an inflammatory response via secreted IL-8. BFT-induced release of latent TGF-beta may facilitate the subsequent repair of the injured epithelium, following its activation by proteases from neighbouring cells. Variation in cytokine responses by colonic epithelial cells in vivo could be an important determinant in the development of mucosal disease and symptoms in response to ETBF.

Bacteroides fragilis↗
PubMed1999

Dictated versus database-generated discharge summaries: a randomized clinical trial.

BACKGROUND: Hospital discharge summaries communicate information necessary for continuing patient care. They are most commonly generated by voice dictation and are often of poor quality. The objective of this study was to compare discharge summaries created by voice dictation with those generated from a clinical database. METHODS: A randomized clinical trial was performed in which discharge summaries for patients discharged from a general internal medicine service at a tertiary care teaching hospital in Ottawa were created by voice dictation (151 patients) or from a database (142 patients). Patients had been admitted between September 1996 and June 1997. The trial was preceded by a baseline cohort study in which all summaries were created by dictation. For the database group, information on forms completed by housestaff was entered into a database and collated into a discharge summary. For the dictation group, housestaff dictated narrative letters. The proportion of patients for whom a summary was generated within 4 weeks of discharge was recorded. Physicians receiving the summary rated its quality, completeness, organization and timeliness on a 100-mm visual analogue scale. Housestaff preference was also determined. RESULTS: Patients in the database group and the dictation group were similar. A summary was much more likely to be generated within 4 weeks of discharge for patients in the database group than for those in the dictation group (113 [79.6%] v. 86 [57.0%]; p < 0.001). Summary quality was similar (mean rating 72.7 [standard deviation (SD) 19.3] v. 74.9 [SD 16.6]), as were assessments of completeness (73.4 [SD 19.8] v. 78.2 [SD 14.9]), organization (77.4 [SD 16.3] v. 79.3 [SD 17.2]) and timeliness (70.3 [SD 21.9] v. 66.2 [SD 25.6]). Many information items of interest were more likely to be included in the database-generated summaries. The database system created summaries faster and was preferred by housestaff. Dictated summaries in the baseline and randomized studies were similar, which indicated that the control group was not substantially different from the baseline cohort. INTERPRETATION: The database system significantly increased the likelihood that a discharge summary was created. Housestaff preferred the database system for summary generation. Physicians thought that the quality of summaries generated by the 2 methods was similar. The use of computer databases to create hospital discharge summaries is promising and merits further study and refinement.

Abstracting and Indexing↗
PubMed1999

Choroidal tuberculosis diagnosed by polymerase chain reaction. A clinicopathologic case report.

PURPOSE: To show the use of the polymerase chain reaction (PCR) in a granulomatous choroidal lesion to support a diagnosis of tuberculosis. DESIGN: Observational case report. TESTING: Nucleic acid target amplification of a choroidal specimen using PCR for detection of Mycobacterium tuberculosis was tested. MAIN OUTCOME MEASURES: Positive nucleic acid target amplification for M. tuberculosis in the ocular sample was measured. RESULTS: PCR was positive for M. tuberculosis with appropriate negative controls. CONCLUSIONS: PCR was thought to be a useful supportive technique in the diagnosis of choroidal tuberculosis.

Choroid Diseases↗
PubMed1998

Production of proinflammatory cytokines and inflammatory mediators in human intestinal epithelial cells after invasion by Trichinella spiralis.

Epithelial cells are the first point of host contact for invasive intestinal pathogens and may initiate mucosal inflammatory responses via production of proinflammatory cytokines and mediators. The aim of the present study was to investigate in vitro the initial invasion of a parasitic nematode (Trichinella spiralis), to measure the early production of specific epithelial cytokines and inflammatory mediators after invasion, and to compare these responses with those to invasive bacteria. Monolayers of human colonic epithelial cell lines (HT29, T84, and Caco-2) were infected by T. spiralis or Listeria monocytogenes. Bile-activated infective larvae of T. spiralis invaded and migrated into the epithelial cell monolayers, leaving trails of dead cells. Transmission electron microscopy studies of damaged cells along the trail showed a progressive increase in size, disruption of cell membranes, loss or dilution of cytoplasmic proteins, and swelling of mitochondria and nuclei. However, no nuclear fragmentation was observed. With reverse transcription-PCR and an enzyme-linked oligonucleotide chemiluminescent assay, mRNA transcripts of interleukin-1beta (IL-1beta), IL-8, and epithelial neutrophil-activating peptide 78 were shown to increase in epithelial cells invaded by T. spiralis or L. monocytogenes, but only L. monocytogenes elicited increased inducible nitric oxide synthase (iNOS) mRNA. No increase in tumor necrosis factor alpha or transforming growth factor beta mRNA was seen after T. spiralis invasion. Increased levels of IL-8 were also released from the basolateral surfaces of infected monolayers as detected by sandwich enzyme-linked immunosorbent assay. Induction and secretion of proinflammatory cytokines in epithelial cells after nematode or bacterial invasion may initiate the acute inflammatory response of the small intestine. The upregulation of iNOS in bacterial infections may contribute to mucosal defense and may also be associated with subsequent cell death, whereas different mechanisms appear to operate after nematode invasion.

Animals↗
PubMed1998

Coexpression of gastrin and gastrin receptors (CCK-B and delta CCK-B) in gastrointestinal tumour cell lines.

BACKGROUND: The peptide hormone gastrin is a recognised growth factor for gastrointestinal (GI) tumour cells. Carboxyamidated gastrins bind to the cell surface gastrin/cholecystokinin B (CCK-B) receptor which can be expressed as either a normal or a truncated isoform (delta CCK-B). AIMS: To compare gastrin gene expression with delta CCK-B and total CCK-B (both isoforms) gene expression in both GI and non-GI tract derived human tumour cell lines. METHODS: Total RNA was extracted and gene expression was assayed by the reverse transcription-polymerase chain reaction followed by Southern blotting and hybridisation with specific oligo probes. RESULTS: Gastrin was expressed by 5/5 gastric and 7/8 colorectal cell lines. Coexpression of gastrin CCK-B isoform was found in 80% of gastric and 75% of colorectal cell lines. Non-GI cell lines, with the exception of a lymphoblastic leukaemia cell line, showed no coexpression. The truncated receptor, delta CCK-B, was shown in 3/5 gastric and 5/8 colorectal cell lines and was always coexpressed with gastrin. CONCLUSIONS: The truncated gastrin receptor, delta CCK-B, is coexpressed with gastrin in 8/13 GI tumour cell lines. Gastrin and CCK-B receptor isoforms may be involved in maintaining autocrine/paracrine growth pathways in GI cancer cells.

Blotting, Southern↗