DNA aneuploidy and histologic dysplasia in long-standing ulcerative colitis. A 10-year follow-up study.

PURPOSE: This study was designed to assess the presence of DNA aneuploidy and mucosal dysplasia, respectively, in 63 patients with long-standing ulcerative colitis. METHODS: The DNA content in colonic biopsies was investigated, using a flow cytometry method, and compared with conventional histology. Patients were subsequently followed each or every second year with colonoscopy and histology. A second flow cytometry examination to monitor the DNA pattern was performed after 10 years. RESULTS: Initially, abnormal DNA pattern (i.e., aneuploidy) was found in 13/63 (21 percent) patients. The colonic mucosa was flat in 10, polypoid in 1, and tumor infiltrated in 2. Eight of the 10 aneuploid cases with a flat mucosa showed no signs of histologic dysplasia. In one of two cases with simultaneous aneuploidy and low-grade dysplasia, a carcinoma Dukes B was found at subsequent colectomy. On the other hand, dysplasia (one low grade and one high grade) without aneuploidy was found in two patients at the initial investigation. After 10 years, 13 had been colectomized, 11 had died (7 noncolitis related), and 3 were lost to follow-up. In the remaining 36 living patients with intact colorectum, no case of histologic dysplasia, but 6 cases of DNA aneuploidy were discovered at the initial investigation. Of the six aneuploid cases, one was later reclassified as diploid and one consisted of an aneuploid adenomatous polyp (removed by polypectomy). At follow-up 10 years later, 3/4 of the other aneuploid cases showed repeated abnormal DNA pattern, now together with histologic low-grade dysplasia (in flat colon mucosa). The 30 patients with initially normal DNA patterns were all still diploid at re-examination 10 years later, but 2 now revealed low-grade dysplasia histologically. CONCLUSIONS: DNA aneuploidy in chronic ulcerative colitis seems to be stable and it may precede histologic dysplasia by many years. It appears to be an additional marker for detecting neoplastic transformation in ulcerative colitis.

Early effects of preoperative irradiation upon the cell cycle composition in rectal adenocarcinomas. A flow-cytometric DNA investigation.

The DNA patterns were studied by means of flow cytometric analysis in 43 rectal adenocarcinomas. Ploidy level and cell cycle distribution were related to clinical stage and histopathology. The frequency of grossly aneuploid tumours and tumours with multiple aneuploid cell populations increased with more advanced clinical stages and with the degree of dedifferentiation. In 15 cases the DNA pattern was studied before and after preoperative irradiation. The ploidy level was not affected by irradiation. A pronounced increase in the proportion of G2 cells was found after irradiation. This G2 blockage was proportional to the amount of S-phase cells before irradiation. Since following irradiation the proportion of S-phase cells was low and the proportion of G1 cells unchanged, the existence of a high fraction of resting G1 tumour cells can be assumed.

Flow-cytometric DNA analysis as a means for early detection of malignancy in patients with chronic ulcerative colitis.

A new approach to the problem of monitoring patients with chronic ulcerative colitis is presented and discussed in connection with a case report. When annual colonoscopies are performed, biopsies are taken for histopathological examination and DNA measurements are made using flow-cytometric techniques (FCM). Using the latter approach, gross chromosomal aberrations indicating malignant transformation in a cell population may be detected. In a 46 year old man with a long history of ulcerative colitis, an area with slight mucosal dysplasia at light microscopy was accompanied by two aneuploid cells lines - that is, colonic mucosa cells with an abnormal amount of DNA in the nuclei. An operation one year later revealed a 5 X 2 mm large adenocarcinoma in the corresponding area of the colon. We suggest that flow-cytometric techniques can be used as a complement to already practised methods for monitoring the colorectal mucosa in colitic patients for the early detection of malignancy.

Villous dysplasia. An ominous histologic sign in colitic patients.

Preoperative biopsies and colectomy specimens from 40 patients with long-standing ulcerative colitis (of whom 20 had adenocarcinoma at colectomy) were searched for the presence of villous adenomatous changes with or without cellular dysplasia. Villous adenomatous changes were found in available preoperative punch biopsies in nearly 70 per cent of the patients with carcinoma, but in none of the preoperative punch biopsies from the 20 patients without cancer. Only three of the preoperative biopsies from patients with carcinoma showed severe dysplasia, and also one of the 20 colitic patients without carcinoma. The mucosal tip in villous adenomatous changes was usually covered by columnar epithelium without dysplasia. In preoperative punch biopsies from patients with long-standing ulcerative colitis, the presence of structures compatible with villous adenoma--even those without cellular dysplasia--should be considered an ominous histologic sign and, thus, an indication for panproctocolectomy.

Early detection of malignancy in ulcerative colitis. A flow-cytometric DNA study.

The cellular DNA content in colorectal biopsies from 51 patients with chronic ulcerative colitis has been studied with flow-cytometric technique. The DNA pattern has been related to the duration of the disease, and to histopathology. Aneuploid cell lines were found in eight patients, two of which had histopathologically verified malignancies. Aneuploid cell lines were found in 2% of biopsies from mucosa classified as normal, in 10% of biopsies with inflammation, hyperplasia or atrophy, in 25% of polyps and in biopsies with dysplasia and in seven of eight biopsies from adenocarcinomas. Four of 27 cases with 11 to 20 years duration of disease, and 4 of 10 with more than 20 years disease, had aneuploid cell lines. Combined with colonoscopy the flow-cytometric technique can routinely be used in the follow-up of patients with ulcerative colitis in screening for malignant cell lines.

Prospective, randomized trial of selective proximal vagotomy with ulcer excision and partial gastrectomy with gastroduodenostomy in the treatment of corporeal gastric ulcer.

In a prospective, randomized trial, selective proximal vagotomy with complete ulcer excision was compared with partial gastrectomy with gastroduodenostomy for the treatment of primary corporeal gastric ulcer in 30 patients. The results were evaluated by clinical follow-up examinations at regular intervals, endoscopy, and blood tests. The mean follow-up period was 3 years for both operations. There was no mortality. Postoperative complications, ulcer recurrence rates, the overall clinical results (Visick classification), and blood test results were similar in the two groups. Three of 15 selective proximal vagotomy patients were classified grade IV due to recurrent ulcer (two patients) or dumping (1 patient), and 2 of 14 partial gastrectomy patients as were classified grades III (epigastric pain) and IV (recurrent ulcer). Considering the risk for late symptoms of impaired resorption and gastric cancer after partial gastrectomy, the similar results with selective proximal vagotomy and partial gastrectomy justify further trials of selective proximal vagotomy with ulcer excision for treatment of corporeal gastric ulcer.

Ploidy and proliferation patterns in colo-rectal adenocarcinomas related to Dukes' classification and to histopathological differentiation. A flow-cytometric DNA study.

The cellular DNA pattern in 66 colo-rectal adenocarcinomas was studied by means of flow-cytometric DNA analysis. The degree of ploidy and the proportion of cells in S-phase were related to the clinical stage according to Dukes' classification and to the histological differentiation. Multiple cell populations were found in about 60 per cent of the tumours but more frequently in advanced clinical stages. According to the DNA index the cell populations were bimodally distributed with one peak in the diploid-peridiploid region and one peak in the tri- to tetraploid region. In the second group there was a higher frequency of more advanced tumours as compared to the first. The proportion of cells in S-phase was higher in pure diploid tumour cell populations of all clinical stages as compared to normal mucosa but lower as compared to peridiploid and aneuploid cell populations with high DNA index. High as well as low S-phase values may occur in all clinical stages, but a significant higher mean value was found for Dukes' C compared to Dukes' B tumours. Distant metastases occur at all DNA indices and with various S-phase values. In conclusion, tumours of different clinical stages and histological differentiation may be subdivided according to DNA index, to the existence of single or multiple cell populations and to the proportion of cells in S-phase. The biological significance of this subdivision can only be evaluated by means of clinical follow-up.

Adriamycin, CCNU, and 5-fluorouracil in patients with advanced gastrointestinal cancer.

Forty-one patients with measurable advanced gastrointestinal malignancy were treated with a combination of Adriamycin (doxorubicin), 5-FU (5-fluorouracil), and CCNU (1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea). The response to therapy was evaluated every eight weeks. In addition to standard clinical examinations and laboratory tests to evaluate toxicity, 31 patients were asked to rank both the expected and experienced negative effects of the treatment. Although both patients with hepatocellular cancer and the patient with a soft tissue sarcoma responded to the regimen, only 1 of 38 patients with adenocarcinoma had a favourable response. The clinically measurable toxicity, principally to the gastrointestinal tract and bone marrow, was relatively mild. However, the patients' subjective evaluation of their treatment made the side effects appear more significant. The results suggest that this regimen deserves further evaluation in patients with hepatocellular cancer and soft tissue sarcomas.

Electron microscopic studies of serially sectioned cat spinal alpha-motoneurons. IV. Motoneurons innervating slow-twitch (type S) units of the soleus muscle.

Two intracellularly stained cat alpha-motoneurons of the soleus-S type (Burke et al., '74) were studied ultrastructurally. The architecture and synaptology of the cell body and proximal parts of the dendrites were analyzed from a long series of consecutive sections (Conradi et al., '79). Only few of the dendrites had a base diameter exceeding 10 micrometer. The proportion of the membrane surface of the cell body and dendrites covered by boutons was 40-45% and 50-80%, respectively. Out of this, 15-20% was constituted by S-boutons and 70-75% by F-boutons in both regions. In contrast to the situation in the gastrocnemius FR-neurons (Kellerth et al., '79) the large boutons of the C-type showed no clustering around the dendritic roots and were absent on the axon hillock. In addition, the M-boutons of dorsal root origin were more sparse on the proximal parts on the dendrites of the soleus-S neurons.

Studies of some twitch and fatigue properties of different motor unit types in the ankle muscles of the adult cat.

Contractile responses of motor units in the gastrocnemius, soleus and pretibial flexor muscles of adult cats were elicited by intracellular stimulation of motoneurones. The motor units were classified into types FF, FR and S (Burke et al. 1971) and their responses to the same stimulation patterns as those used in a previous investigation of whole muscles (Hammarberg and Kellerth 1975 a) were studied. The duration of motoneurone afterhyperpolarization was short in both the fast twitch FF and FR units; it was longer in the soleus S units than in the S units of the pale muscles. Twitch time-to-peak was less than 30 ms in the FF and FR units, but exceeded 40 ms in the S units. Soleus S units were slower than S units of the pale muscles. Potentiation was observed in the gastrocnemius units, but not in the soleus S units. A short rest allowed fatigued extensor units of the FF and FR types to regain some contractile strength. This was less evident in the S units which, on the other hand, were extremely resistant to fatigue. Differences in response patterns between corresponding motor unit types of the flexor and extensor muscles were observed. A few fast twitch units were identified in the slow soleus muscle.

The postnatal development of some twitch and fatigue properties of single motor units in the ankle muscles of the kitten.

Contractions of single motor units in the gastrocnemius, soleus and pretibial flexor muscles were elicited by intracellular stimulation of the innervating motoneurones. Kittens being 1, 2, 6 and 10 weeks of age were used. The aim was to establish the pattern of postnatal differentiation of the adult motor unit types (Burke et al. 1973, 1974, Hammarberg and Kellerth 1975 b). In the developing type S units of the soleus muscle the mean value for contraction time showed a transient decrease during the early postnatal period, but the twitch half-relaxation time (HRT), the duration of motoneurone post-spike afterhyperpolarization (AHP) and the susceptibility to fatigue remained virtually unchanged during the age period studied. In the units of the developing fast twitch muscles the contraction time, HRT and susceptibility to fatigue were significantly different from those of the soleus units already at 1 week of age. At 6-10 weeks of age the contractile characteristics typical of the adult type FF and FR units were attained. The AHP duration gradually decreased up to 10 weeks of age, and it was then considerably shorter than both at 1 week of age and in the adult stage.

The postnatal development of some twitch and fatigue properties of the ankle flexor and extensor muscles of the cat.

The isometric responses of the medial gastrocnemium (MG), soleus (SOL) and anterior tibial (TA) muscles to single shocks and different modes of repetitive stimulation were studied in kittens of varying postnatal ages and in adult cats. The postnatal decrease in time-to-peak and half-relaxation time of the twitch contractions were similar for the MG and TA muscles and adult values were attained at around 6-7 weeks of age. The SOL muslce displayed a transient decrease in contraction time during the first postnatal weeks, followed later by a slowing towards adult values. The susceptibility to fatigue during iterative stimulation was smallest in the SOL at all ages studied, and usually largest in TA. It changed only little for the MG and SOL postnatally while increasing markedly for the TA up until 6-7 weeks of age. Tetanic contraction resulted in similar depressions in contractile tension of all three muscles in the youngest kittens, but the SOL displayed a greater ability to recover from this depression than the MG and, in particular, the TA muscles. Tetanus resistance increased postnatally and adult responses were attained at 6-7 weeks of age.