Identification of sentinel nodes in patients with colon cancer.

AIMS: Lymphatic mapping and sentinel node biopsy entails better staging in malignant melanoma and breast cancer and we used this technique in patients with colon cancer to possibly improve detection of lymphatic spread. METHODS: Thirty patients with invasive adenocarcinomas were investigated. The tumour status in identified sentinel node(s) was compared with the status in all other harvested regional nodes for each patient. Patients were followed at regular visits for more than 30 months. RESULTS: Sentinel nodes were identified in all cases, either per-operatively (28 cases) or at dissection of the formalin-fixed specimen (2 cases). The sentinel nodes were diagnostic for the entire lymphatic field in 28 patients and the false negative rate was 2/12. In four cases, the sentinel nodes were the only metastatic nodes. After at minimum 30 months, three patients had died of colon cancer metastases. CONCLUSION: This method improved the identification of patients with lymph-node metastases, especially those with only few metastatic nodes. Patients dying from metastatic disease had lymph-node metastases at diagnosis.

Helicobacter pylori strain types and risk of gastric cancer: a case-control study.

The aim of this novel endoscopy clinic-based case-control study was to explore the influence of different Helicobacter pylori strain types on the risk of gastric adenocarcinoma using isolated bacterial strains, tissue samples, and sera. We included 72 cases with gastric adenocarcinoma and 324 age- and sex-matched controls. Histological characterization, culture, molecular typing of H. pylori genes by PCR (cagA/vacA), conventional IgG ELISA, and immunoblotting (Western blot) for the CagA and VacA proteins were performed. With four tests combined, H. pylori infection was detected in 57 (79%) cases and 213 (66%) controls. A positive association between H. pylori infection and gastric cancer risk was found [odds ratio (OR), 2.1; 95% confidence interval, 1.1-3.9]. Type I (OR, 1.8), intermediate (OR, 2.0), and type H (OR, 0.2) strains of H. pylori presented different serum antibody levels and different levels of association with gastric cancer. Our case-control study, based on molecular characterization and serology, provides further evidence that infection by more virulent strains of H. pylori and the presence of antibodies toward the CagA protein can be used as markers for an increased risk of gastric adenocarcinoma and that the strain types of H. pylori could be used in the future to determine disease outcome.

13C-urea breath test results in pigs challenged with Helicobacter pylori or other urease producing bacteria.

The gastric bacterial flora and its influence on the 13C-urea breath test (UBT) for detection of Helicobacter pylori infection was studied in a pig model. Seven SPF minipigs were used. H. pylori or a mix of other urease positive bacteria were administered orally. UBT, serum and biopsies for histology and culture were collected. Our results show that UBT is not specific for H. pylori in pigs as the gastric bacterial flora is responsible for the high UBT values observed. Furthermore, the Ellegaard Göttingen SPF minipigs are not useful in an animal model for H. pylori studies.

Complete sequence of p53 gene in 20 patients with lung cancer: comparison with chemosensitivity and immunohistochemistry.

In this study the entire p53 complementary DNA has been sequenced in 20 non-small cell lung carcinomas (NSCLC) and the results correlated with chemosensitivity, immunohistochemistry and clinical data. Ten patients had mutations in p53, 8 missense mutations and 2 nonsense mutations. The method discovered two mutations never described previously and two other mutations that have never been described before in connection with NSCLC tumours. Chemosensitivity data, according to a short-term assay (FMCA), indicated that tumours with p53 mutation were more resistant to cisplatin and cyclophosphamide. Immunohistochemical studied demonstrated a 70% concordance between over-expression of p53 protein and mutation in p53. No conclusions or trends could be drawn from the immunohistochemical studies of Bcl-2 and Bax.

Is the circumferential resection margin a predictor of local recurrence after preoperative radiotherapy and optimal surgery for rectal carcinoma?

OBJECTIVE: Circumferential resection margin (CRM) involvement has been correlated with a high risk of developing local recurrence. The aim of this study was to examine the prognostic significance of the CRM involvement after curative resection of rectal cancer in patients treated with preoperative radiotherapy and postoperative chemotherapy where indicated. METHOD: All patients with rectal cancer treated in a regional central unit from 1996 to 2004 were identified. A surgical resection was performed on 257 patients, and in 229 of these this was assessed as potentially curative. The CRM was examined in all patients. A CRM of < or = 1 mm was considered positive. RESULTS: A positive margin was seen in 19 (8%) patients. At a median follow up of 40 months, only four (1.7%) patients had developed local recurrence, one of whom had a positive CRM. In the four patients the tumour was 5 cm or less from the anal verge. There were no significant differences regarding local recurrence and survival between CRM positive and negative tumours. CONCLUSION: Rectal cancer managed by combined radiochemotherapy and surgery resulted in a low positive CRM rate and a low local recurrence rate. An involved CRM was not a predictor of local recurrence.

Magnetic resonance imaging for preoperative staging of rectal cancer in clinical practice: high accuracy in predicting circumferential margin with clinical benefit.

OBJECTIVE: The aims were to determine agreement between staging of rectal cancer made by magnetic resonance imaging (MRI) and histopathological examination and the influence of MRI on choice of radiotherapy (RT) and surgical procedure. METHOD: In this retrospective audit, preoperative MRI was performed on 91 patients who underwent bowel resection, with 93% having total mesorectal excision. Tumour stage according to mural penetration, nodal status and circumferential resection margin (mCRM) involvement was assessed and compared with histopathology. RESULTS: Five radiologists interpreted the images. Overall agreement between MRI and histopathology for T stage was 66%. The greatest difficulty was in distinguishing between T1, T2 and minimal T3 tumours. The accuracy for mCRM (MRI) was 86% (78/91),with an interobserver variation between 80% and 100%. In the 13 cases with no agreement between mCRM and pCRM (pathological), seven had long-term RT and nine en bloc resections, indicating that the margins initially were involved with an even higher accuracy for mCRM. Preoperative short-term RT was routine, but based on MRI findings, choice of RT was affected in 29 cases (32%); 17 patients had no RT and 12 long-term RT. The surgical procedure was affected in 17 cases (19%) with planned perirectal en bloc resections in all. CRM was involved (< or = 1 mm) in 14.7% of the 34 cases in which MRI had an effect upon choice of RT and/or surgery compared with 8.8% of the remaining 57 cases where it had no impact. CONCLUSION: Magnetic resonance imaging predicted CRM with high accuracy in rectal cancer. MRI could be used as a clinical guidance with high reliability as indicated by the low figures of histopathologically involved CRM.

[Influence of indole-3-alkanecarboxylic acids on glucose utilization in rats]. / Einfluss von Indol-3-alkan-carbonsäuren auf die Glukoseverwertung bei Ratten

The influence of various indole-3-alkane-carboxylic acids (indole-3-propionic acid, indole-3-butyric acid and derivatives of this substance) on some parameters of carbohydrate, fat and insulin metabolism were studied in normal and experimentally induced diabetic rats. Earlier it had been shown, by Schillinger and Loge (1973) using rat liver slices for in vitro studies and adrenalectomized rates for in vivo experiments that indole-3-butyric acid and its analogues investigated suppress gluconeogenesis from pyruvate and that these substances have a hypoglycaemic effect when administered orally at doses of 50-250 mg/kg. Indole-3-propionic acid, indole-3-butyric acid, 1-methyl-indole-3-butyric acid, 2-methyl-indole-3-butyric acid and 1-carboxy-6-fluor-1,2,3,4-tetrahydrocarbazol, administered orally at doses up to 1000 mg/kg, had either only a slight or no hypoglycaemic effect at all in intact and streptozotocin-diabetic rats. The suppression of gluconeogenesis observed in vitro thus only has an appreciable effect on blood glucose in vivo when the capacity of the liver to synthetize glucose is reduced. In normal rats indole-3-propionic acid and indole-3-butyric acid led to a deterioration of glucose tolerance following i.v. glucose loading and to a reduction of 14C-U-glucose oxidation. An increase in the concentration of serum free fatty acids (FFA) measured after treatment with indole-3-butyric acid and its 1- and 2-methylated derivatives was not--in accordance with a theory of an interference of the glucose and fatty acid utilization published in literature--considered to be the primary cause since the reduced formation of 14CO2 from radioactive glucose is not normalized when the increased FFA content is reduced by an inhibitor of lipolysis (5-methyl-isoxazol-3-carboxylic acid)...

Adult coeliac disease within a defined geographic area in Sweden. A study of prevalence and associated diseases.

An epidemiologic study of coeliac disease in a geographically defined area of Sweden showed that the prevalence was 95.5/10(5) inhabitants aged 15 years or more. The highest prevalence, 178/10(5) inhabitants, was found in the age group 65-74 years. The lowest prevalence, 39/10(5) inhabitants, was found in patients aged 15-24 years. Among the associated diseases an especially high incidence of associated thyroid disease was observed: thyrotoxicosis occurred in 5.0% and hypothyroidism in 5.8% of the patients.

Azodisal sodium in the treatment of ulcerative colitis. A study of tolerance and relapse-prevention properties.

One hundred sixty patients intolerant of or allergic to sulfasalazine (Salazopyrin, Azulfidine) participated in an open tolerance study of azodisal sodium (Dipentum). More than 4 of every 5 patients tolerated azodisal sodium well, but 12.5% of patients stopped medication because of diarrhea. Even after 7 patients who had also experienced diarrhea when taking sulfasalazine were excluded, there still remained a group of patients (9.8%) who had to discontinue azodisal sodium because of diarrhea. Apart from this, only minor side effects occurred. No serious drug-related changes were seen in hematologic or biochemical parameters. Male fertility appeared to be unaffected. One hundred two patients, who were in clinical and sigmoidoscopic remission, took part in a double-blind, placebo-controlled maintenance trial. Of these, 23.1% of the patients treated with azodisal sodium and 44.9% of the patients treated with placebo had a clinical and sigmoidoscopic relapse during a 6-mo trial period (p = 0.02). Azodisal sodium appears to be an effective agent for the maintenance treatment of ulcerative colitis.

Crohn's disease after in-utero measles virus exposure.

BACKGROUND: An epidemiological association between Crohn's disease and measles virus exposure in early life has been suggested in case-control studies. METHODS: To determine absolute risk estimates for in-utero measles virus exposure and Crohn's disease, maternity charts for all 25000 deliveries at University Hospital, Uppsala, between 1940-49 were reviewed: four cases of measles infection in the mother during pregnancy were identified. The children and two of their mothers were interviewed and case records reviewed. Three offspring had undergone multiple intestinal resections; tissue from these cases were examined by routine histology, and for measles-virus nucleoprotein antigen by immunohistochemistry and immunogold electronmicroscopy. FINDINGS: Three of the four children had Crohn's disease. In each the disease was preceded by recurrent, antibiotic-resistant pneumonia. They had extensive ileal and colonic disease; two patients required intravenous feeding. The only offspring to have had measles as a child did not develop Crohn's disease. Measles virus antigen was detected in foci of granulomatous and lymphocytic inflammation in all children with Crohn's disease. INTERPRETATION: The data indicate that exposure of mothers to measles virus in utero is a risk factor for Crohn's disease in their children. Exposure at this time may lead to persistent infection, or modify the response to infection in later life, leading to persistence of measles virus.

Prevalence of Campylobacter pylori in an unselected Swedish population of patients referred for gastroscopy.

A survey of the prevalence of Campylobacter pylori in an unselected series of 117 patients referred for gastroscopy showed that 49.6% of the patients were infected. In 87% of infected patients both the antrum and the body of the stomach were involved. Duodenal infection occurred in only 9% of 102 studied patients. C. pylori could usually be demonstrated in mucosa showing signs of active chronic inflammation, whereas it was rarely found in histologically normal mucosa. The acridine orange staining method for demonstration of bacilli correlated well with bacteriology; identical results were obtained in 92.8% of the samples.

Rectal cancer: evaluation of staging with endosonography.

PURPOSE: To investigate whether endosonography is reliable in making radiation therapy decisions in rectal cancer, with possible downstaging taken into consideration. MATERIALS AND METHODS: Ninety patients (52 men, 38 women; median age, 69 years) with rectal adenocarcinoma underwent endosonography within 2 weeks before surgery and radiation therapy (performed in 54 patients). The tumor invasive edge was used for radiation therapy decision making. RESULTS: The local stage was accurately assessed in 65 patients (39 with and 26 without irradiation). The tumor invasive edge was accurately assessed in 63 patients. Overstaging was present in 19 patients; the tumor had grown almost through the muscularis propria in six. The invasive edge (P = .1) and lymph node status were overstaged more often in the patients with than in the patients without irradiation. Tumor was understaged in eight patients: The invasive edge did not penetrate but there was budding beyond the muscularis propria in five; the invasive edge penetrated the muscularis propria in two. In seven of the eight patients, growth beyond the muscularis propria was smaller than the endosonographic resolution. Three patients with understaged, nonirradiated tumors developed pelvic recurrence. None of the patients with irradiation and none of the 16 patients without irradiation but with correct assessment developed pelvic recurrence. CONCLUSION: Preoperative irradiation decision making on the basis of endosonographic findings is uncertain. Downstaging after preoperative irradiation must be considered.

Psoriasis patients have highly increased numbers of tryptase-positive mast cells in the duodenal stroma.

We have shown that the number of tryptase-positive mast cells in the duodenal mucosa in psoriasis is increased and that a subgroup of psoriasis patients showed elevated levels of antibodies to gliadin (some of whom also had increased lymphocytes in the duodenal epithelium). Duodenal biopsy specimens from 37 patients with psoriasis (eight mild, 13 moderate and 16 severe) and 22 patients with irritable bowel syndrome (IBS) were examined regarding the presence of tryptase + mast cells. Intraepithelial infiltration by lymphocytes was evaluated and scored from 0 to 3. Patients with psoriasis had 131 +/- 58 mast cells/mm2 (mean +/- SD) and those with IBS 28 +/- 18. Only in four of the 37 psoriasis patients was the number within the range of that in the IBS group. There were no signs of stromal inflammation except in one psoriasis patient. No relationship was found between degree of severity of psoriasis and number of mast cells. In 25 of the 37 specimens from psoriasis patients there was no increase in intraepithelial lymphocytes, whereas seven showed a slight increase (score 1-2) and five a moderate increase (score > or = 2-3). The number of tryptase + mast cells was similar in patients with or without increased intraepithelial lymphocytes. The number of mast cells showed no relation to the presence or absence of antibodies to gliadin. We hypothesize that there are at least two types of abnormalities in the duodenal mucosa in psoriasis, one type that is present in most psoriasis patients and characterized by an increase in mast cells and eosinophils, and another that is present in a subgroup of patients with antibodies to gliadin and an increased number of duodenal intraepithelial lymphocytes. The mechanisms underlying the increase in the number of mast cells and its relevance are not yet known.

Production of antibodies to gliadin by peripheral blood lymphocytes in children with celiac disease: the use of an enzyme-linked immunospot technique for screening and follow-up.

Problems in the diagnosis of celiac disease are that a long time is needed between challenge with gluten and the appearance of the typical diagnostic morphologic signs in gut mucosa. Furthermore, local immunity to gliadin is only slowly and often incompletely mirrored by serum IgA anti-gliadin antibody (AGA) levels. It is known that a local IgA-associated immune response in the gut may be better and more quickly mirrored by an increase of circulating IgA-producing cells against the immunogen than by IgA serum antibodies. We have therefore used an enzyme-linked immunospot (ELISPOT) assay to enumerate IgA AGA spot-forming cells (SFC) in peripheral blood in 82 children with suspected celiac disease or with other gastrointestinal symptoms. The numbers of IgA AGA SFC/10(6) mononuclear cells were markedly increased in 17 patients with untreated (and later biopsy-verified) celiac disease compared with healthy children, children with nonceliac disease, and patients treated for celiac disease (p < 0.0001). In 20 children with celiac disease the numbers of IgA AGA SFC increased rapidly (p < 0.0001) after gluten challenge. As early as 2 wk after gluten challenge, 15/20 of these patients had abnormal levels of IgA AGA SFC, 6/20 patients had increased levels of serum IgA AGA, and 7/20 had IgA anti-endomysium antibodies. Our results indicate that analysis of IgA AGA production in peripheral blood cells may in further clinical studies provide a sensitive method for the diagnosis of celiac disease after a short time of gluten challenge.

Vulvar papillomatosis, aceto-white lesions, and normal-looking vulvar mucosa evaluated by microscopy and human papillomavirus analysis.

BACKGROUND AND OBJECTIVES: To assess the presence of human papillomavirus (HPV) DNA in vulvar papillomatosis, since some women with this complaint have symptoms associated with HPV infection, such as itching, burning, and dyspareunia. GOAL: To reassure the patients that they do not have a transmissible viral disease, by excluding a HPV origin of their condition. STUDY DESIGN: Vulvar biopsy specimens from 22 females with vulvar papillomatosis, from 10 females with prominent aceto-white vulvar lesions, and from 14 healthy controls were analyzed histologically for signs of HPV infection and by in situ hybridization and polymerase chain reaction for the presence of HPV DNA. RESULTS: Specimens from women with vulvar papillomatosis showed some histological signs associated with HPV infection, but no koilocytotic atypia or dysplasia and thus resembled normal-looking vulvar mucosa. Aceto-white lesions frequently displayed histological features suggestive of HPV infection, including dysplasia. HPV DNA was detected in 6 of 10 patients with aceto-white lesions, but only in 1 of 22 patients with papillomatosis and in no healthy controls. CONCLUSIONS: According to our results, HPV DNA, which is generally found in cervical lesions and subclinical infections, is not present in vulvar papillomatosis even though symptoms associated with HPV infection are frequent complaints.

The skin and the gut in psoriasis: the number of mast cells and CD3+ lymphocytes is increased in non-involved skin and correlated to the number of intraepithelial lymphocytes and mast cells in the duodenum.

The aim of this work was to study tryptase+ mast cells and CD3+ T lymphocytes in non-involved skin in psoriasis and their possible relation to mast cells and lymphocytes in the duodenal mucosa. Skin biopsy specimens were obtained from 43 patients with psoriasis of variable severity and from 10 healthy subjects. Compared with the reference subjects, the number of mast cells in non-involved skin was clearly increased, most markedly in the papillary dermis. The increase was present both in mild, moderate and severe psoriasis. CD3+ lymphocytes were increased in non-involved skin in moderate and severe psoriasis. Patients with an increased number of duodenal intraepithelial lymphocytes had significantly more mast cells in non-involved skin than those without such an increase, and there was a significant correlation between the number of mast cells in non-involved skin and score for intraepithelial lymphocytes. However, when the 14 patients with increased intraepithelial duodenal lymphocytes were excluded-as they may represent a separate type of psoriasis-another type of correlation between the skin and the duodenal mucosa was found, namely a highly significant inverse correlation between the number of CD3+ lymphocytes in non-involved skin and the number of duodenal mast cells, which is highly elevated in psoriasis. The results might indicate an interplay between skin and intestinal mast cells and lymphocytes in a hitherto unknown way.

Histopathologic examination of penile epithelial lesions is of limited diagnostic value in human papillomavirus infection.

BACKGROUND AND OBJECTIVES: Guidelines for the histopathologic diagnosis of human papillomavirus (HPV) infection have been drawn from changes seen in cervical specimens not necessarily applicable to penile epithelium. GOAL: To evaluate histopathologic examination as a means of diagnosing HPV infection of the male genital tract. STUDY DESIGN: Ninety-two consecutive male patients seen at the sexually transmitted diseases clinic. Twelve had condyloma acuminatum, and 80 had papular lesions, macular lesions, or both. Fifteen men without signs of HPV infection served as controls. Biopsy specimens were evaluated morphologically by light microscopy, and HPV DNA detection was performed by in situ hybridization and polymerase chain reaction. RESULTS: All acuminate lesions were HPV DNA positive with in situ hybridization. Forty papular and/or macular lesions harbored HPV DNA, 28 (35%) of them positive with in situ hybridization and the other 12 (15%) positive with polymerase chain reaction. Morphologic signs attributed to HPV infection were found in HPV-positive and HPV-negative penile lesions, as well as in normal epithelium. In papular and macular lesions, the only criterion associated with HPV DNA positivity was neoplastic changes, which was present in 16 (40%) HPV DNA-positive specimens, compared to 4 (10%) HPV DNA-negative specimens (P < 0.01). Of the 16 lesions with neoplasia, 15 (94%) had detectable HPV DNA of a potentially oncogenic type. CONCLUSIONS: Histopathologic signs of HPV infection other than neoplasia seem to be of limited value. Detection of the infectious agent, in this case HPV, should be the gold standard for the diagnosis as it is for other infectious diseases. The strong association between neoplasia and potentially oncogenic HPV types makes this issue even more important.

Increased expression of the transcription factors CCAAT-enhancer binding protein-beta (C/EBBeta) and C/EBzeta (CHOP) correlate with invasiveness of human colorectal cancer.

Regulation of cell differentiation is most often impaired in malignant tumors and may represent a key mechanism for the progression of the disease. CCAAT-enhancer binding protein (C/EBP) is a family of transcription factors involved in the regulation of embryonic gut development in rodents, which has also been detected in various malignancies, e.g., liposarcomas and breast and ovarian epithelial tumors. We studied the relationship between C/EBP and tumor histology (Duke's invasive stage and pathological grade) in colorectal cancer. Immunoblotting techniques were used on microdissected fresh frozen tumor specimens, and expression of C/EBPalpha, C/EBPbeta and C/EBPzeta (CHOP) was analyzed in addition to that of the cell-cycle regulator p53 and the proliferation marker PCNA. Expression of C/EBPbeta (LAP isoforms) was markedly increased in all tumors compared with normal colon mucosa. Although the inter-patient variability was large, we found that LIP, the isoform of C/EBPbeta known to inhibit transcription, was expressed at higher levels in Duke's stage B tumors compared with Duke's stage A, whereas Duke's C tumors had the lowest LIP expression. A similar relationship was seen for CHOP. The cell-cycle regulator gene p53 was the only factor that clearly correlated with pathological grade: a decrease in p53 expression was demonstrated. Our data suggest that genetic and cellular events involving C/EBPbeta and CHOP are important for tumor invasion and that these events do not appear to be related to the pathological grade of the tumor.