Expression and localisation of matrix metalloproteinases and their natural inhibitors in fistulae of patients with Crohn's disease.

BACKGROUND: Fistulae are a troublesome complication of Crohn's disease but little is known of the final effector molecules responsible for matrix degradation. Although matrix metalloproteinases (MMPs) have been strongly implicated in tissue injury in Crohn's disease, their role in fistula formation is unknown. AIM: To determine the expression pattern of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in fistulae of patients with Crohn's disease. PATIENTS AND METHODS: Resected fistula specimens were obtained from patients with Crohn's disease (n = 11) and classified according to the predominant histological features-that is, acute versus chronic inflammation. Patients with fistulae due to other diseases (n = 9) and normal colon (n = 5) served as controls. MMP and TIMP protein expression was measured by single or double labelled immunohistochemistry, and mRNA expression by in situ hybridisation. MMP activity was measured by gelatin zymography. RESULTS: Compared with normal colon, strong MMP-3 expression was consistently observed in fistulae in Crohn's disease, irrespective of the stage of inflammatory activity. MMP-3 transcripts and protein were localised in large mononuclear cells and fibroblasts. MMP-9 transcripts and protein were expressed in granulocytes and only in fistulae with acute inflammation. Staining for MMP-1 and MMP-7 was weak and negative for MMP-10, whereas MMP-2 was equally expressed in normal colon and fistulae. TIMP-1, TIMP-2, and TIMP-3 expression was low in all samples. Similar expression patterns were found in fistulae of the disease control group. Fistulae also expressed active MMP-2 and MMP-9, as measured by gelatin zymography. CONCLUSION: MMP-3 and MMP-9 are markedly upregulated in intestinal fistulae and may contribute to fistula formation through degradation of the extracellular matrix, irrespective of the underlying disease process.

Screening for dysplasia and TP53 mutations in closed rectal stumps of patients with ulcerative colitis or Crohn disease.

BACKGROUND: Patients who undergo colectomy due to intractable chronic inflammatory bowel disease (IBD) may keep a closed rectal stump for several years, which may be at increased risk of malignant transformation owing to residual inflammatory activity. We examined a hospital series of patients with ulcerative colitis or Crohn colitis to describe the clinical, endoscopical and histological features of the closed rectal stump and to screen for dysplasia and mutations in the TP53 tumour suppressor gene. METHODS: During rigid proctoscopy, rectal mucosal biopsy specimens and rectal lavage fluid were collected from 42 patients. Biopsy specimens were examined histologically, and genomic DNA extracted from frozen biopsies and lavage fluid was analysed for mutations in TP53 exons 4-9. RESULTS: The median disease duration was 8.5 years (range 1.3-34 years). No endoscopic or histological signs of dysplasia or carcinoma were seen and no mutations in the TP53 gene were detected in any biopsy or lavage fluid specimens. Histological moderate to severe mucosal inflammation was present in 78% (33/42) of the patients, however, and rectal stump involution was noted in 43% (18/42). CONCLUSION: No signs of malignancy or premalignant degeneration were detected in this prospective series of IBD patients with a closed rectal stump. Although this is reassuring for patients, the presence of moderate to severe inflammation in the majority of rectal stumps indicates a role for adjuvant molecular markers to improve colorectal cancer surveillance on this subgroup of IBD patients.

Chronic idiopathic anal pain: analysis of ultrasonography, pathology, and treatment.

PURPOSE: This study was undertaken to analyze whether intra-anal ultrasound examination, anorectal physiologic evaluation, and histopathologic examination in patients with chronic idiopathic anal pain presented any common features and whether the results of different treatment modalities correlated with these findings. METHODS: Eighteen patients who met the criteria for chronic idiopathic anal pain were studied. All had an intra-anal ultrasound examination and a complete anorectal physiologic evaluation. In a selected group of patients, ultrasound-guided biopsy samples were taken from pathological areas in the internal and external sphincter. Treatment consisted of analgesics only in four patients, 0.2 percent nitroglycerin ointment in four, and ultrasound injection of botulin (botulinum toxin, Botox) into the intersphincteric space in nine. Two patients, including one who was previously treated with botulin, ultimately had a colostomy. RESULTS: Four patients were managed satisfactorily on analgesic treatment under the guidance of the hospital's pain clinic. Nitroglycerin ointment resulted in temporary pain relief in one of four patients. Injection of botulin resulted in a permanent improvement in four patients, a temporary improvement in one patient, and no effect in four patients. Two patients had a colostomy, resulting in complete pain relief. The effect or lack of effect of nitroglycerin ointment and botulin was not related to changes in anal pressure. CONCLUSION: Chronic idiopathic anal pain is a condition of unknown origin for which no proven therapy exists. As in other syndromes based on muscular dystonia, some patients may benefit from injection of botulin.

[Positron-emission tomography with gamma camera in patients with colorectal and anal cancer. Preliminary results]. / Positron-emissionstomografi med gammakamera hos patienter med kolorektal cancer og analcancer. Praeliminaere resultater.

Metastasectomy after the primary surgical treatment of colorectal cancer may prolong survival in patients with resectable recurrences. Dedicated 18F-FDG-PET is of growing interest as a diagnostic tool in the staging and diagnosis of recurrent disease. We wanted in our departments to investigate whether a dual-head gamma camera with coincidence detection could serve as a substitute for the dedicated PET scanner. Twenty consecutive PET scans of 18F-FDG were performed by means of an Adac VertexPLUS gamma camera in 14 patients with colorectal cancer and four patients with anal cancer with known or suspected recurrences. Evaluation of the patients were, however, according to the routine in the department. In fourteen patients one or more abnormal foci were detected by PET, nine of these were verified by biopsy and one by renewed CT scan. In the four patients with negative PET the findings were in agreement with later biopsies. In one patient PET was "falsely" negative with regards to other processes than a solitary metastasis to be removed, as he six months later revealed dissemination confirmed by biopsy. Four patients with positive PET findings and elevated CEA remain to be evaluated. We conclude that PET with a coincidence detection gamma camera in this preliminary study seems to have the same possibilities to stage and diagnose recurrent anal- and colorectal cancer as the dedicated PET scanner.

The effect of Buserelin versus conventional antiandrogenic treatment in patients with T2-4NXM1 prostatic cancer. A prospective, randomized multicentre phase III trial. The "Danish Buserelin Study Group".

A prospective, randomized multicentre phase III trial was undertaken to compare the effectiveness and safety of Buserelin, a gonadotropin-releasing hormone analogue (GnRHa), with conventional antiandrogenic treatment in patients with painful metastases from T2-4NXM1 prostatic cancer. Seventy-two patients received Buserelin, 22 received estrogens and 46 were subjected to orchiectomy. The trial was completed one year after allocation of the patients. No significant differences as regards suppression of testosterone or survival were found in favour of one of the three treatment modalities. The performance index improved significantly both during the first months of treatment with Buserelin and following orchiectomy. No detectable improvement of performance index was seen during treatment with estrogens. Treatment with estrogens also failed to alleviate pain or general symptoms of cancer. Tolerability, safety and compliance of Buserelin was although administered intranasally clearly evidenced as palliation of advanced symptomatic cancer and the efficacy and sideeffects were fully comparable to those following orchiectomy.

Carcinoid tumour of the breast.

A case of primary carcinoid in the female breast is reported. The patient was treated by segmental mastectomy and is without local recurrence and signs of carcinoid tumour at any other site after 3 years of observation.

Breath hydrogen analysis in patients with ileoanal pouch anastomosis.

The possible influence on functional outcomes of hydrogen production in the ileoanal pouch after restorative proctocolectomy was investigated by means of lactulose H2 breath tests. Eight of 15 patients had significant increases in breath hydrogen after 10 g lactulose. One patient declined to participate in further investigations, the remaining seven responders had no evidence of small bowel bacterial overgrowth after glucose H2 breath tests. The ability to produce hydrogen by anaerobic fermentation of lactulose in the pouch was unrelated to the age of the patients or of the pouch. Seven of eight responders had successive breath tests after ingestion of lactulose 20 g and wheat starch 100 g. Five of seven had significant increases after lactulose but none after wheat starch. The overall function of the pouch continence, spontaneity of defecation, and 24 hour stool frequency was significantly better in responders than in non-responders. The absence of H2 production of 100 g wheat starch may indicate either increased absorption or defective fermentation.

The value of quantitative DNA flow cytometry of testicular fine-needle aspirates in assessment of spermatogenesis: a study of 137 previously maldescended human testes.

In order to assess the suitability of DNA flow cytometry of fine-needle aspirates for quantifying spermatogenesis, the results from DNA flow cytometry were compared to histological evaluation of testicular biopsies taken concomitantly from 171 previously maldescended testes. In 137 of 171 cases, sufficient material for flow cytometric as well as histological evaluation was obtained. Histological analysis of surgical biopsy specimens revealed spermatogenesis including the spermatid stage in 117 of the 137 gonads. In six of the 117 gonads no haploid cells were found using flow cytometry. On the other hand, surgical biopsies failed to reveal spermatogenesis in five cases in which the corresponding aspirates contained haploid cells. Both methods therefore seem equally sensitive in detection of spermatogenesis. Other types of histological patterns also corresponded to distinct DNA histograms. Thus, in 11 of 12 cases with Sertoli-cell-only pattern in all tubules, at least 95% of the cells had a diploid DNA content. Furthermore, predominance of tubules with maturation arrest at the primary spermatocyte level corresponded to an increased proportion of tetraploid cells. When compared to surgical biopsy, DNA flow cytometry of testicular fine-needle aspirates is a more objective, easy and rapid method, which is more convenient for the patient. This study has indicated that DNA flow cytometry is a suitable method of quantitative assessment of spermatogenesis. One of the first target groups might be men with azoospermia. In such men, DNA flow cytometric analysis of fine-needle aspirates and surgical biopsy are apparently of equal sensitivity in detecting gonads with spermatogenesis. We conclude that DNA flow cytometry may become an alternative method for the quantification of spermatogenesis.

Histological grade, DNA ploidy and mean nuclear volume as prognostic factors in prostatic cancer.

Seventy-nine consecutive patients with prostatic cancer diagnosed between 1979 and 1983 were classified with respect to histological grade according to Shelley et al. (23), DNA ploidy (diploid, tetraploid and aneuploid), and mean nuclear volume (mnv) of the cancer cells. Nine patients were still alive at the time of analysis (minimum observation time 7.5 years). Univariate analysis showed that histological grade (p = 0.004), DNA ploidy (p = 0.0002) and mnv (p = 0.009) all had a significant influence on crude survival. A Cox proportional hazards model was used to analyse the prognostic value of these factors and patient age. However, when the two strongest of these four factors, histological grade (p = 0.0025) and DNA ploidy (p = 0.0035), were corrected for, age (p = 0.07) and mnv (p = 0.22) had no independent impact on crude survival.

[Sucralfate versus placebo in the treatment of abdominal wound abscesses]. / Sukralfatpulver versus placebo i behandling af spaltede abdominale søarabscesser.

Twenty-seven patients were divided at random into two groups following abscess formation and complete reopening of the abdominal wound in a prospective double blind investigation to study the effect of topical sucralfate on the wound healing rate in secondary abdominal wounds. Fifteen patients were treated with sucralfate powder and 11 patients with placebo while one patient was excluded. The two groups were comparable as regards age, sex and factors predisposing to abscess formation. In the sucralfate group the median healing rate was 2.8 cm2/24 hours as compared with 2.4 cm2/24 hours in the placebo group (p = 0.92, Mann-Whitney test). From this investigation, it is concluded that topical application of sucralfate powder does not appear to increase the rate of healing in secondary abdominal wounds after incision of a wound abscess.

[Partial spleen resection using a stapler]. / Partiel miltresektion ved hjaelp af haeftemaskine.

At emergency laparotomy after a fall bleeding from injuries to the upper pole and from lacerations involving the hilum of the spleen were found in a 2 1/2-year-old girl. Partial splenectomy using a linear stapler allowed preservation of the lower pole with intact vessels. Observed peroperatively for 10 minutes, the splenic remnant retrained a macroscopically normal appearance. Blood transfusion was avoided and drainage postoperatively comprised no more than 25 ml serosanguineous fluid. Provided intact splenic capsule can be used for the line of resection, partial splenectomy using a linear stapler seems to be a safe method.

The prognostic value of histological grading and mean nuclear volume in human prostatic cancer.

Fifty-eight consecutive patients diagnosed with prostatic cancer at transurethral resection in the period 1979-1983 were classified using histological grade according to Shelley, and using an unbiased estimate of mean nuclear volume. We find that both histological grade and mean nuclear volume appear to be significantly associated with survival. In eight of nine patients still alive after 89-130 months mean nuclear volume at diagnosis was in normal range and the histological grade showed a highly differentiated cancer in eight of the nine cases. These findings suggest highly differentiated cancer and normal range of mean nuclear volume may be important for long-term survival. In patients with advanced disease requiring endocrine treatment both histological grading and mean nuclear volume estimates failed to show any prognostic properties with regards to time to progression and time of survival. In patients not subjected to endocrine treatment histologic grade, but not mean nuclear volume, was found to be significantly associated with survival. Subtle changes in disease progression may be diagnosed early by repeated biopsies due to the accuracy and high reproducibility of mean nuclear volume measurements; however, a single estimate of mean nuclear volume has not been shown to offer advantage over histological grade with respect to prognostic properties.

DNA distributions in maldescended testes: hyperdiploid aneuploidy without evidence of germ cell neoplasia.

Fine-needle aspiration biopsies and surgical biopsies were obtained from maldescended testes of 149 consecutive men. The aspirates were subjected to quantitative DNA flow cytometry and the surgical biopsy to histological evaluation. From more than 80% of the gonads, sufficient material was obtained for both examinations. A significant hyperdiploid cell population with a mean DNA index of 1.23 (range 1.17-1.31) was found in six gonads. Hyperdiploid aneuploidy was found in gonads without, as well as with, complete spermatogenesis. In none of the six cases did the surgical biopsy show evidence of early testicular neoplasia by morphology or by immunohistochemical methods with antibodies against carcinoma in situ. This indicates that aneuploidies in maldescended testes do not necessarily indicate malignancy. It may be speculated that hyperdiploid aneuploidy is related to the development of preneoplastic lesions.

Prevalence of carcinoma in situ and other histopathological abnormalities in testes of men with a history of cryptorchidism.

The incidence of invasive testicular cancer is increased in men with a history of cryptorchidism. Previous studies based on relatively small series indicated that the risk of carcinoma in situ of the testis also is increased in these men. In our study 500 consecutive men 20 to 30 years old, who were previously admitted to a department of surgery with the diagnosis of testicular maldescent, were asked to participate in a screening for carcinoma in situ of the testis. Of the men 300 consented to testicular biopsy. The biopsies were evaluated by light microscopy for carcinoma in situ and other histopathological abnormalities. Carcinoma in situ was diagnosed in 5 patients (1.7%, 95% confidence limits 0.5 to 3.9%). However, the true risk of carcinoma in situ might be higher, since 2 men who had been treated for testicular cancer before they were offered biopsy were excluded from the study. Advanced spermatogenesis, including the spermatid stage in all tubules, was found in biopsy specimens from only 37% of the men. In 80% of these specimens even the number of late spermatids was decreased. Thus, our study, based on a large number of testicular biopsies from an unselected group of men with testicular maldescent, provided further evidence that these men have an increased risk for carcinoma in situ of the testis. Our data combined with the results of other Scandinavian studies indicate that the true prevalence of carcinoma in situ in men with a history of cryptorchidism is approximately 2 to 3%. Additionally, we confirmed that spermatogenic function is severely impaired in maldescended gonads. Invasive testicular cancer can be prevented if the neoplasm is detected at the stage of carcinoma in situ. In our opinion the magnitude of prevalence of carcinoma in situ found in men with a history of cryptorchidism justifies that these men should be offered testicular biopsy when they reach adulthood.

Stereological estimation of mean nuclear volume in prostatic cancer, the reproducibility and the possible value of estimations on repeated biopsies in the course of disease.

The reproducibility of stereological estimates of mean nuclear volume by using the principle of volume estimation of particles of arbitrary shape is investigated together with the possible prognostic value of the estimates in the course of advanced prostatic cancer. Repeated transurethral resection of the prostate was performed 14-93 months after the first resection in 14 of 85 consecutive patients in whom prostatic cancer was newly diagnosed in the period 1979-1983. Twelve of these fourteen patients needed endocrine treatment because of progression of the metastatic disease. The reproducibility of the estimations was excellent as the variation of the method was less than 1% when the estimation of the mean nuclear volume was repeated at random on the same specimens after a time-interval of 2 months. The progression of the disease was evidenced by a significant histological upgrading and by a highly significant increase of the mean nuclear volume from the first to the second transurethral resection of the prostate.

Clinical applicability of an amino-terminal radioimmunoassay for determination of parathyroid hormone.

With a radioimmunoassay using hPTH 1-34 for antibody production, for radioiodination and as a standard, hPTH 1-34 was detectable (detection limit 40 pg/ml) preoperatively in peripheral blood in 14 of 29 patients with hyperparathyroidism, but in no controls. In all patients with parathyroid adenoma and detectable hPTH 1-34 preoperatively, the values fell after parathyroid surgery. Contrastingly, three of four patients with diffuse parathyroid hyperplasia and two of three with normal parathyroid glands showed a rise in hPTH 1-34 postoperatively, which was concomitant with very low serum calcium levels. In studies of hPTH 1-34 in central venous blood (3 patients), levels were detectable in all samples, but not in simultaneously drawn peripheral blood. Values for hPTh 1-34 in central blood correlated to PTH determined from a bovine assay, but the peripheral samples showed no correlation. The low levels of circulating N-terminal immunoreactivity in peripheral blood make this assay inapplicable for routine diagnostic purposes. These low levels are due not only to low secretion rates, but also to rapid peripheral metabolization.

Intravesical therapy of noninvasive bladder tumors (stage Ta) with doxorubicin: initial treatment results and the long-term course.

A total of 64 consecutive patients with noninvasive bladder tumors (stage Ta) between 1977 and 1983 underwent treatment with intravesical instillation of doxorubicin. The total response rate was 85 per cent and 57 per cent of the patients had a complete response after 16 weekly instillations. Half of the patients with a complete response had recurrences within 12 to 16 months. Ten per cent of all patients had an invasive tumor during the observation period. We conclude that doxorubicin can render some patients with noninvasive bladder tumors (stage Ta) free of disease but there currently is nothing to indicate that the same results could not be obtained by transurethral resection.

Dihydrotestosterone measured in core biopsies from prostatic tissues.

Dihydrotestosterone (DHT) levels were measured and related to the content of DNA in 10- to 15-mg tissue samples obtained by punch biopsies from the prostatic gland. The aim of the study was to provide an opportunity to detect the variation of dihydrotestosterone (DHT) in prostatic tissue on endocrine manipulations in phase III studies of metastatic prostatic cancer. DHT was assayed in ranges of 27.9 to 102.9 pg/ml or when related to DNA from 0.57 to 4.00 pg/micrograms DNA by modifying a commercial testosterone/dihydrotestosterone kit. Preliminary clinical results from nine patients with prostatic cancer and eight with benign hyperplasia showed a significantly lower DHT/DNA ratio in prostatic cancer. The predictive value of the histological grading was improved by estimates of the weighted mean nuclear volume. The trend that mean nuclear volume of prostatic cancer exceeded the mean nuclear volume of hyperplasia was clear, and a significantly larger nuclear variation was demonstrated in the malignant specimens. Neither the DHT/DNA ratio nor the mean nuclear volume appears to be sufficient to guide the handling of individual patients, but the described methods enable us by repeated measurements to follow the course of disease and response to treatment.