Microsatellite instability and hMLH1/hMSH2 expression in Barrett esophagus-associated adenocarcinoma.

BACKGROUND: Microsatellite instability (MSI) has been documented in malignancies associated with hereditary nonpolyposis colon carcinoma and in sporadic malignancies of the colon, stomach, and endometrium. In these malignancies, MSI is associated with defects in the DNA mismatch repair enzymes hMSH2 and hMLH1. Defects in these enzymes result in a phenotype characterized by instability of multiple microsatellite repeat sequences throughout the genome. This study sought to determine the prevalence of MSI in 80 primary Barrett esophagus-associated adenocarcinomas (BEAd) and to examine the relation of MSI with the clinical and pathologic features of the tumors. METHODS: Eighty BEAd were evaluated for the presence of MSI by using the microsatellite markers BAT25, BAT26, D10S219, D10S541, and D10S551. These tumors also were evaluated for immunohistochemical expression of hMSH2 and hMLH1. RESULTS: High levels of MSI were not found in any of the tumors examined. Furthermore, immunohistochemical expression of hMSH2 and hMLH1 was retained in all cases evaluated. Evidence of low level MSI was found in 16% of tumors. In none of these tumors, however, was MSI present in more than two of five loci. The presence of MSI did not correlate with patient age, tumor stage, degree of differentiation, or with patient survival. CONCLUSIONS: High level MSI and loss of hMLH1/hMSH2 expression is uncommon in BEAd. A subset of BEAd demonstrate low level MSI. The presence of low level MSI was not associated with the clinicopathologic features of the tumors examined.

Allelic loss of 10q23, the PTEN tumour suppressor gene locus, in Barrett's oesophagus-associated adenocarcinoma.

PTEN is a putative tumour suppressor gene located on chromosome band 10q23. Mutations in PTEN have been identified in numerous human malignancies, including cancers of the brain, endometrium, ovary, and prostate. In this study, we screened 80 Barrett's oesophagus-associated adenocarcinomas (BOAd) for loss of heterozygosity (LOH) at 10q23, using the microsatellite markers D10S541, D10S219, and D10S551. Tumours demonstrating LOH were then screened for the presence or absence of PTEN mutations. LOH at one or more loci was identified in 17/80 (21%) cases. In none of these cases did we detect mutations in PTEN. The presence of LOH did not correlate with patient age, tumour stage, degree of differentiation, presence of perineural or vascular invasion, or overall survival. We conclude that LOH at chromosome 10q23 is uncommon in BOAd, is not associated with mutations in the PTEN tumour suppressor gene, and does not correlate with the clinical or pathologic features of these tumours. It is possible that PTEN is inactivated through other mechanisms in BOAd.

Cardiac chamber-specific alterations of ANP and BNP expression with advancing age and with systemic hypertension.

The present study determined cardiac chamber-specific alterations of the expression of the atrial and brain natriuretic peptide (ANP and BNP) genes with a small increase in age beyond adulthood and with systemic hypertension of intermediate duration. The expression distributions of these genes was determined using in situ hybridization in the right and left atria (RA and LA), and the right and left ventricles (RV and LV) in Wistar Kyoto rats (WKY) and age-matched Spontaneously Hypertensive rats (SHR) at ages 6 months (adult) and 8 months (advanced-age beyond adulthood). In all rat groups, both genes were expressed (ANP > BNP) in the LA and LV, and were not expressed in the RA and RV. The genes were expressed in the LA in all rat groups; the ANP, but not the BNP, expression increased with advancing age and with superimposed hypertension. They were expressed in the LV of the advanced-age WKY, adult and advanced-age SHR, but not in the adult WKY. The ANP mRNA labeling in the LA was diffuse and interspersed with dense accumulations, whereas BNP labeling was diffuse. The labeling of both genes in the form of sparse clusters was seen in the LV of the advanced-age SHR. Our study showed that ANP and BNP expression in left heart chambers increased with a small increase in age, with hypertension of intermediate duration, and with modest left ventricular hypertrophy. The chamber-specific expression distribution could be due to special groups of cardiac cells, or to local chamber-specific factors.

Immunohistochemical evidence of loss of PTEN expression in primary ductal adenocarcinomas of the breast.

Germline mutations in PTEN, encoding a dual-specificity phosphatase on 10q23.3, cause Cowden syndrome (CS), which is characterized by a high risk of breast and thyroid cancers. Loss of heterozygosity of 10q22-24 markers and somatic PTEN mutations have been found to a greater or lesser extent in a variety of sporadic component and noncomponent cancers of CS. Among several series of sporadic breast carcinomas, the frequency of loss of flanking markers around PTEN is approximately 30 to 40%, and the somatic intragenic PTEN mutation frequency is <5%. In this study, we analyzed PTEN expression in 33 sporadic primary breast carcinoma samples using immunohistochemistry and correlated this to structural studies at the molecular level. Normal mammary tissue had a distinctive pattern of expression: myoepithelial cells uniformly showed strong PTEN expression. The PTEN protein level in mammary epithelial cells was variable. Ductal hyperplasia with and without atypia exhibited higher PTEN protein levels than normal mammary epithelial cells. Among the 33 carcinoma samples, 5 (15%) were immunohistochemically PTEN-negative; 6 (18%) had reduced staining, and the rest were PTEN-positive. In the PTEN-positive tumors as well as in normal epithelium, the protein was localized in the cytoplasm and in the nucleus (or nuclear membrane). Among the immunostain negative group, all had hemizygous PTEN deletion but no structural alteration of the remaining allele. Thus, in these cases, an epigenetic phenomenon such as hypermethylation, -ecreased protein synthesis or increased protein degradation may be involved. In the cases with reduced staining, 5 of 6 had hemizygous PTEN deletion and 1 did not have any structural abnormality. Finally, clinicopathological features were analyzed against PTEN protein expression. Three of the 5 PTEN immunostain-negative carcinomas were also both estrogen and progesterone receptor-negative, whereas only 5 of 22 of the PTEN-positive group were double receptor-negative. The significance of this last observation requires further study.

Epididymal cystadenomas in von Hippel-Lindau disease.

OBJECTIVES: Epididymal cystadenomas (ECs) are frequently found in association with von Hippel-Lindau disease (VHL), but little has been reported about their sonographic appearance. We review the sonographic appearance of ECs, the relationship of ECs to other manifestations of VHL, and the specific genetic mutations associated with ECs. METHODS: Fifty-six male patients with VHL were examined with scrotal sonography and physical examination as part of a larger screening program for VHL. The head of the epididymis was measured in two planes on sonography and compared with age-matched normal controls. All VHL patients with palpable epididymal abnormalities or enlargement (more than two standard deviations) of the head of the epididymis on ultrasound were considered positive for EC. RESULTS: Thirty of 56 (54%) male patients with VHL demonstrated a unilateral (n = 10; 33%) or bilateral (n = 20; 67%) solid abnormality in the head of the epididymis suggestive of EC. Sonographic appearances ranged from a solid mass with multiple tiny cysts to an almost completely solid mass. The most common appearance was a 15- to 20-mm solid mass with small cystic components. Dilated efferent ductules were seen within the testicle in 7 men, evidently a result of chronic obstruction. There was no association between the clinical subtype of VHL and the presence of ECs (P > 0.10, chi square). Mutations resulting in a truncated gene product were associated with the development of ECs but the association did not reach statistical significance (P = 0.06). CONCLUSIONS: ECs are a common manifestation of VHL in men and exhibit a range of appearances on ultrasound. Sonography can be used to identify ECs and determine the extent of cystic dilation of the rete testes. The benign course of ECs and the usual absence of clinical symptoms favor a conservative approach to their management.

Background subtraction in technetium-99m-MAG3 renography.

UNLABELLED: Correction represents a potential source of error in estimating split renal function and camera-based clearances. The purpose of this study was to determine which of five background options and four time intervals was associated with the least error for 99mTc-mercaptoacetyltriglycine (MAG3). METHODS: Fifteen single-kidney patients were imaged supine after 111-370 MBq (3-10 mCi) 99mTc-MAG3 injection. A phantom kidney was drawn on the 2-3-min images, approximately equal in size to the solitary kidney and used for all time intervals. Counts in the phantom and native kidneys were calculated using manual inferior and lateral regions of interest (ROIs), automated elliptical and perirenal background ROIs and no background correction at various time intervals (1-2, 1-2.5, 1.5-2.5 and 2-3 min) postinjection. With optimal background correction, counts and the relative function in the phantom kidney should be zero. The error was measuring by estimating both the relative function and absolute function expressed as the percent injected dose in the phantom kidney. RESULTS: The percent injected dose in the phantom kidney as well as the error in measuring relative function were significantly greater than zero for the inferior background correction and the no background correction options at all time intervals, p < 0.05. The percent dose in the kidney and the error associated with the lateral, elliptical and perirenal ROIs were not significantly different from zero. CONCLUSION: Regardless of time interval, the greatest error was associated with no background correction. The inferior ROI consistently underestimated the background correction and probably should not be used for 99mTc-MAG3. There was no significant difference between errors generated using the lateral and automated ROIs, although automated ROIs are probably more reproducible for sequential studies.

Small (< or = 3-cm) renal masses: detection with CT versus US and pathologic correlation.

PURPOSE: To determine the sensitivities of computed tomography (CT) and ultrasound (US) for detection and characterization of surgically verified small renal lesions. MATERIALS AND METHODS: Twenty-one patients with von Hippel-Lindau disease or hereditary papillary renal cancer underwent CT and US before partial nephrectomy or enucleation; 205 renal masses were removed (92% were <3 cm). Detection rates and accuracy of CT and US in the characterization of renal morphology were correlated with lesion size. RESULTS: CT and US detection rates for lesions of 0-5 mm were respectively 47% and 0%; 5-10 mm, 60% and 21%; 10-15 mm, 75% and 28%; 15-20 mm, 100% and 58%; 20-25 mm, 100% and 79%; and 25-30 mm, 100% and 100%. Among the lesions 10-35 mm, 80% and 82% were correctly characterized with CT and US, respectively. CONCLUSION: A substantial proportion of lesions under 1 cm were not detected with either modality. Neither CT nor US was superior in the characterization of lesions 3 cm or less. CT and particularly US screening studies in patients with von Hippel-Lindau disease should be interpreted cautiously because missed or mischaracterized small renal lesions are a frequent problem in these patients.

Intestinal peptide YY: ontogeny of gene expression in rat bowel and trophic actions on rat and mouse bowel.

The purpose of this study was twofold: 1) to characterize the profile of colonic peptide YY (PYY) gene expression in rats and 2) to examine for potential trophic effects of PYY on the intestine in rats and mice. Expression of PYY mRNA (analyzed by Northern blotting and in situ hybridization) and PYY (analyzed by high-performance liquid chromatography and radioimmunoassay) was detected initially at day 17 of gestation in colonic extracts of Sprague-Dawley and Fischer rats. Expression of colonic PYY mRNA increased until 7 days of age and remained at its highest level (approximately twofold greater than the adult level) through the end of the nursing period. After weaning (21 days of age), PYY mRNA levels declined quickly to adult levels. Colonic PYY concentrations followed, in a coordinated manner, with some temporal delay after birth, the increase and decrease of its mRNA. Administration of PYY increased the weight and DNA content of the duodenum significantly in nursing rats and adult mice. In mice, PYY treatment also increased weight and DNA content of the ileum and colon. The trophic effects of PYY were dose related, peptide specific, and independent of species and sex. From these findings, we hypothesize that PYY plays an important role in intestinal development and dietary adaptation.

Effect of aging on neurotensin-stimulated growth of rat small intestine.

The proliferative activity of gut mucosa is altered with aging; the potential for the aged gut to respond to trophic stimuli is not known. The purpose of this study was to determine whether there are age-related differences in the effects of the trophic gut peptide neurotensin (NT) on the structure and function of small bowel mucosa. NT (300 micrograms/kg) or saline (control) was injected subcutaneously at 8-h intervals for 5 days in rats of two age groups, young (2 mo) and aged (24 mo). On day 6, rats were killed, and the gut mucosa (proximal and distal small bowel) was scraped, weighed, and analyzed for DNA, RNA, and protein content and for disaccharidase (sucrase and maltase) activity. In a second experiment, the groups of rats and the protocol for NT administration were identical; however, when the rats were killed, the distal gut was removed for histological evaluation of crypt and villus length (mm) and density (no./cm gut segment) and bromodeoxyuridine immunohistochemistry. NT produced significant increases in mucosal growth (wt, DNA, RNA, and protein) in both age groups when compared with age-matched controls; the increase of growth measurements was the greatest in the small bowel mucosa of the aged rats. In addition, NT increased crypt density in both groups; only the aged group treated with NT demonstrated increases in crypt depth and villus height. Specific activities of sucrase and maltase did not change with NT treatment in either of the age groups. We conclude that the proliferative potential of small bowel mucosa is maintained with aging in response to administration of NT.(ABSTRACT TRUNCATED AT 250 WORDS)

Symptomatic interleukin-2-induced cholecystopathy in patients with HIV infection.

OBJECTIVE: This study reports the clinical and radiologic findings in seven patients infected with HIV who had 10 consecutive episodes of symptomatic cholecystopathy induced by infusion of interleukin-2. SUBJECTS AND METHODS: Ten episodes of right upper quadrant pain associated with gallbladder wall thickening were seen in seven of 29 HIV-infected patients who received IV interleukin-2. Patients received 6-18 million IU/day of continuous interleukin-2 infusion for 5 days. Patients with right upper quadrant pain underwent sonographic examinations, which were interpreted prospectively. Medical records and previous sonographic studies were reviewed retrospectively. Follow-up was obtained through outpatient visits and sonography. RESULTS: Right upper quadrant pain during these 10 episodes of cholecystopathy usually developed 4-5 days after starting infusion of interleukin-2. Sonography during that time showed gallbladder wall thickening (mean thickness, 12.4 mm; range, 5-18 mm) and a wide variety of sonographic appearances. Tenderness during sonography was focal in six episodes, diffuse in one, and absent in three. Sludge was identified in one episode; calculi were not seen. Findings on radionuclide biliary scans were normal in three cases. Symptoms abated rapidly in every case after infusion of interleukin-2 was reduced or stopped. No surgery was necessary. When treatment was repeated, three patients had recurrent episodes, with clinical courses and sonographic aberrations showing little variance from the initial episodes. Follow-up sonography in three episodes showed a maximal thickness of the gallbladder wall of 4 mm. No patient had a history or laboratory evidence of intrinsic biliary disease. CONCLUSION: Symptomatic thickening of the gallbladder wall during infusion of interleukin-2 can exactly mimic other forms of acalculous cholecystitis, except that when associated with interleukin-2 the thickening is rapidly reversible and surgery is not required. Radionuclide scans can be useful in clinical decision making. The process appears to be benign, and cessation of interleukin-2 therapy, along with close clinical observation, appears to be the appropriate treatment.

Autoprotection: stimulated tissue repair permits recovery from injury.

Autoprotection is a phenomenon whereby prior exposure to a small dose of a chemical results in protection against a subsequently administered lethal dose of the same compound. While CCl4 autoprotection has been studied the most, it has also been demonstrated for other chemicals. Recent studies indicate that the prevailing concept of decreased bioactivation of the normally lethal dose of CCl4 owing to decreased hepatic microsomal cytochrome P-450 content cannot be supported by direct end points of liver injury such as necrosis. These findings suggest a pivotal role for hepatocellular division and tissue healing processes stimulated by the protective dose in the mechanism of autoprotection. Augmentation of hepatocellular regeneration and tissue repair, stimulated by the protective dose, appears to permit timely recovery and restoration of hepatic structure and function. In the absence of the protective dose, hepatocellular division is substantially deficient and it occurs too late to tip the delicate balance between recovery from injury and progression of massive injury in favor of recovery. Abolition of autoprotection by colchicine antimitosis, under conditions where metabolism and disposition of CCl4 are not altered, is supportive of this concept. Selective colchicine antimitotic suppression of the early phase of hepatocellular division and tissue repair induced by a low dose of CCl4 results in progression of toxic liver injury, leading to hepatic failure and mortality. Studies have shown that pretreatment with phenobarbital results in postponed low-dose CCl4-stimulated cell division by 24 hours, which accordingly postpones the optimal autoprotection.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of phenobarbital and mirex pretreatments on CCl4 autoprotection.

Male Sprague-Dawley rats maintained on either normal diet (N) or on a diet containing phenobarbital (PB; 225 ppm) or mirex (M; 10 ppm) for 15 days received either corn oil or 1 single administration of a protective dose of CCl4 (0.3 ml/kg, po) on day 16. At 24, 48, 72, 96, or 144 hr after the protective dose, a high dose of CCl4 (5 ml/kg, po) was administered to rats of all the groups, and they were observed for 14-day lethality. In a second experiment, in rats maintained on N, PB, or M diet, liver microsomal cytochromes P-450, aminopyrine demethylase, and aniline hydroxylase were measured at various time points after the administration of the protective dose of CCl4. Serum aspartate transaminase, alanine transaminase, and sorbitol dehydrogenase elevations and histopathological changes observed under a light microscope were used as toxic end points to assess hepatotoxicity. Autoprotection was 100% when the high dose was given at 24 hr after the protective dose in N rats, whereas it was only 55% in PB- or M-pretreated rats. For later time points of 48, 72, and 96 hr, autoprotection was only around 50% in N rats, whereas it was almost 100% in PB- and M-pretreated rats. When the high dose was administered at 144 hr after the protective dose, autoprotection further declined to 25% in N rats and to 75% in M-treated rats, but it remained at 100% in PB-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Triple-phase bone scan findings in aggressive fibromatosis. Before and after radiation therapy.

The authors present comparative triple-phase bone scan findings in three cases of histologically proven aggressive fibromatosis both before (initial evaluation) and after radiation therapy. The purpose of the study was to compare triple-phase bone scan findings in aggressive fibromatosis both before and after radiation therapy and to determine whether any additional physiological information could be obtained. Before radiation therapy, the triple-phase bone scintigraphy demonstrated increased flow and radiotracer pooling in the areas of tumors on dynamic flow and immediate blood pool images, respectively. However, the delayed static images demonstrated variable radiotracer uptake. When compared to preradiation therapy triple-phase bone scan, decreased vascularity was well demonstrated in all three patients after radiation therapy. In addition, it also provided information regarding the changes in the size and extent of tumor, noninvaded underlying bone, and remainder of the skeleton. This additional information can be particularly useful in patients with equivocal or questionable histologic diagnosis especially from small, unrepresentative biopsies.

Rapid plasma clearance of albumin-acrolein adduct in rats.

Protein adducts are used as markers of chemical exposure. Determination of the clearance rate of these adducts from the blood circulation will provide the time frame for their measurement. Radioactive albumin was prepared biosynthetically by repeated intraperitoneal injections of L-[4,5-3H]lysine to a rat. After an affinity purification, an aliquot of this native [3H-lysine]albumin was adducted with 5 mM acrolein. Both the native albumin (A-treated group) and the albumin-acrolein adduct (AAA-treated group) were intravenously injected to separate groups of rats, and the clearance of radioactivity from the plasma was measured as a function of time. At the end of the experiment (33 h after the injection), radioactivity in the whole plasma, and in homogenates of liver, kidney and spleen and their trichloroacetic acid(TCA)-soluble and -insoluble fractions in both A- and AAA-treated groups, was measured. The results, at the initial 11 h after the injection, showed that the radioactivity was cleared from the circulating plasma more rapidly in the AAA-treated group (32% of the injected radioactivity remained) than the A-treated group (52%). At 33 h after the injection, 22% of the injected radioactivity remained in the plasma in the AAA-treated group as compared to 32% in the A-treated group. The whole homogenates of liver and kidney and their corresponding TCA-soluble fractions showed higher radioactivity in the AAA-treated group as compared to the A-treated group. However, the TCA-insoluble fractions from livers and kidneys of the AAA-treated group showed lower radioactivity as compared to the A-treated group. These results indicated that the albumin-acrolein adduct was removed more rapidly from the circulation than the native albumin, and degraded more rapidly by the liver and kidney. There was no preferential removal or degradation of the adducted albumin by the spleen.

Preoperative localization of parathyroid tissue with technetium-99m sestamibi 123I subtraction scanning.

To evaluate the utility of technetium-99m (Tc-99m) sestamibi for visualization of functioning parathyroid tissue, 14 subjects underwent Tc-99m sestamibi 123I subtraction scanning as part of the preoperative evaluation for hyperparathyroidism. Informative scans were obtained in 13 subjects, including 7 patients with recurrent or persistent hyperparathyroidism, and correctly identified the location of the hyperfunctioning parathyroid tissue found at surgery. In all informative patients, hyperparathyroidism was due to adenomatous disease or hyperplasia secondary to renal failure. Successful scans were obtained with glands as small as 220 mg. In the lone patient in whom Tc-99m sestamibi scanning failed to localize hyperfunctioning parathyroid tissue, surgery revealed a 1700-mg hyperplastic parathyroid neoplasm in the neck. In no case did a Tc-99m sestamibi scan suggest parathyroid tissue where there was none. In 1 case, a patient presented with persistent hyperparathyroidism after 1 neck and a second combined neck and mediastinal exploration. Tc-99m sestamibi imaging revealed uptake in the periaortic region, and a 570-mg adenoma was found in the aortopulmonary window. Using only initial studies, prospective evaluation provided a sensitivity of 78.5% and a positive predictive value of 100%. After repeat studies in 5 patients, 2 of 3 patients with initially negative results and technically deficient scans became positive on restudy. Inclusion of these studies increased sensitivity to 93%. Tc-99m sestamibi 123I subtraction scanning appears to be a reliable noninvasive method for preoperative localization of hyperfunctioning parathyroid tissue.

The natural history of renal lesions in von Hippel-Lindau disease: a serial CT study in 28 patients.

OBJECTIVE: Von Hippel-Lindau disease is a multisystem disorder predisposing to renal cysts and cancer. The growth and development of these renal lesions have not been documented previously. We reviewed serial CT scans to determine the rates and patterns of growth of renal lesions associated with von Hippel-Lindau disease. SUBJECTS AND METHODS: Twenty-eight patients with von Hippel-Lindau disease and renal involvement, including the spectrum from simple cysts to solid masses, had follow-up examinations for at least 1 year (mean, 2.4 years; range, 1-12 years) with serial contrast-enhanced abdominal CT. Renal lesions were measured and characterized. Surgical correlation was available in 12 patients. RESULTS: Two hundred twenty-eight lesions (eight lesions per patient) were detected. On the basis of their CT appearance, 168 lesions (74%) were classified as cysts, 18 (8%) as cysts with solid components, and 42 (18%) as solid masses. Among 12 patients with pathologic confirmation, the solid components of cystic lesions and solid lesions almost always contained renal carcinoma. The majority of cysts remained the same size (71%) or enlarged (20%); 9% became smaller or entirely involuted during the follow-up period. Although it is generally presumed that renal cysts are precursors to cancers, the transformation of a simple cyst to a solid lesion was observed in only two patients. Among the 42 solid lesions, all but two enlarged with time, with a mean doubling time of 10 months. CONCLUSION: The renal lesions associated with von Hippel-Lindau disease exhibited wide differences in growth. The majority of renal cysts grew slowly but some involuted. Transition to solid renal cancer was rare among cysts. Complex cystic and solid lesions contained neoplastic tissue that uniformly enlarged. These data may be used to help predict the progression of renal lesions in von Hippel-Lindau disease.