Interaction between haemochromatosis and transferrin receptor genes in different neoplastic disorders.

A number of genes are involved in iron metabolism, including the transferrin receptor (TFR) and haemochromatosis (HFE) genes. In previous investigations an increased risk for neoplastic disease has been observed in individuals homo- and heterozygous for hereditary haemochromatosis. The HFE wild-type gene product complexes with the transferrin receptor (TF) and two different HFE mutations (Cys282Tyr and His63Asp) have been found to increase the affinity of TFR for TF and increase cellular iron uptake. In a recent study we found no associations for HFE and TFR separately, but an interaction between HFE and TFR genotypes in multiple myeloma. Individuals carrying the HFE Tyr282 allele (homo- and heterozygotes) in combination with homozygosity for the TFR Ser142 allele had an increased risk. In the present study the same association was found in breast and colorectal cancer. The odds ratio for all three neoplasms combined was 2.0 (95% CI 1.0-3.8). The risk for neoplastic disease was further increased (OR 7.7, 95% CI = 1.0-59.9) when the analysis was restricted to HFE Tyr homozygotes and compound heterozygotes in combination with TFR Ser homozygosity. Thus, an interaction between HFE and TFR alleles may increase the risk for different neoplastic disorders.

p53 polymorphisms and haplotypes in breast cancer.

Three polymorphisms in the human tumor suppressor gene p53 (BstUI and MspI RFLPs in exon 4 and intron 6 respectively and a 16 bp duplication in intron 3) and their haplotype combinations were studied in patients with breast cancer and controls. A significant increase in the codon 72 BstUI A1 (pro) allele frequency (P = 0.016) and of individuals carrying the pro allele (pro/pro and pro/arg) (OR, 1.47; P = 0.01 4; 95 % CI, 1.08-2.00) was observed in breast cancer. This increase was most pronounced in highly differentiated breast cancer. Significant associations were found only in BstUI and haplotypes containing this polymorphism, which indicates that the codon 72 pro allele may be functionally involved in low malignancy breast cancer. The distributions of genotypic combinations in breast cancer patients and controls were significantly different (P = 0.005). Two BstUI-16 bp-MspI combinations were significantly overrepresented; 2-1, 1-1, 2-2 (OR, 1.61; 95% CI, 1.13-2.30) and 1-1, 2-1, 2-1 (OR, 2.94; 95% CI, 1.37-6.27).

Intra-epithelial lymphocytes. Evidence for regional specialization and extrathymic T cell maturation in the human gut epithelium.

The human gut epithelium is a unique immunological compartment, containing substantial amounts of intra-epithelial lymphocytes (IEL) with unknown functions. In this study we show that distinct and unusual subpopulations of IEL are present at different levels of human intestine. IEL phenotypes in normal jejunum, ileum and colon were compared using immunoflow cytometry and immunohistochemistry. The expression of mRNA for recombination-activating gene-1 (RAG-1) in IEL from all three levels was compared using reverse-transcription polymerase chain reaction, and the morphology of IEL in situ was determined using immunoelectron microscopy. Surface marker profiles of isolated intestinal epithelial cells at all three levels were also investigated. On average the proportion of TCR gamma delta IEL was comparable in jejunum than ileum and colon and varied in phenotype with gut level. CD4-CD8-TCR alpha beta IEL dominated in colon but were absent in jejunum. CD8+ TCR alpha beta IEL were present at all levels but only in jejunum did they constitute the majority of all IEL. CD4+ TCR alpha beta IEL were present in similar frequencies at all levels of the gut. In general, the majority of IEL had an activated phenotype (CD45RO+, alpha E beta 7+). Furthermore, IEL exhibited phenotypes which are rare in peripheral blood. The thymocyte markers CD1a and CD1c as well as the NK cell marker CD56 were expressed on a fraction of TCR alpha beta and TCR gamma delta IEL. A small population of 'null' cells (CD45+ TCR/CD#-CD20-CD14-CD15- cells) was also present at equal proportions along the gut. Jejunal but not colonic IEL expressed RAG-1 mRNA suggesting that extrathymic T cell maturation occurs in the epithelium of small intestine. RAG-1 was expressed in CD2+TCR/CD3- and CD3+/TCR-IEL. Ultrastructurally, IEL often formed small clusters and intimate contacts with epithelial cells, suggesting cell cooperation within the epithelium. Some IEL had pseudopodium-like extensions penetrating the epithelial basement membrane suggesting transmigration. Epithelial cells in small intestine but not colon expressed heat shock protein 60 and HLA-DR. CD1a, CD1b and CD1c were not expressed on intestinal epithelial cells at any level. The distinct surface marker profiles of IEL and epithelial cells along small and large intestine suggest functional regional specialization and are compatible with the hypothesis that TCR alpha beta IEL participate in immune reactions to lumenal antigens while TCR gamma delta IEL perform surveillance of the epithelium.

P53 germ line haplotypes associated with increased risk for colorectal cancer.

Three p53 DNA polymorphisms (BstU I and Msp I restriction fragment length polymorphisms (RFLPs) in exon 4 and intron 6 respectively, and a 16 bp duplication in intron 3) and their haplotype combinations were studied in patients with colorectal cancer and compared with patients with ulcerative colitis and healthy controls. There were only minor differences between patients with ulcerative colitis and controls, the only significant difference was observed in the distribution of BstU I-Msp I haplotypes. When single polymorphisms were studied, a significantly lower frequency of the 16 bp duplication was found in patients with colorectal cancer. The protective effect of the 16 bp duplication was more pronounced in haplotype combinations with the BstU I A1 and Msp I A1 alleles, whereas these alleles in combination with the 16 bp A1 allele (no duplication) were associated with an increased risk for colorectal cancer. The genotypic combination BstU I 2-1, 16 bp 1-I, Msp I 2-1 was found in 8.4% of cases among patients with colorectal cancer and 0.5% of cases in the controls (odds ratio = 18.8). The extended haplotype responsible for the high cancer risk of this genotype appears to be BstU I A1-16 bp A1-Msp I A1. The results of this study indicate that the haplotype approach to the identification of p53 germ line alleles associated with increased susceptibility to cancer is far more powerful than the analysis of single polymorphisms, since the capacity to identify germ line alleles predisposing to cancer should increase with the number of polymorphic sites included in the analysis.

Local recurrence and long-term survival in patients with gastric cancer--analysis of possible impact of clinicopathological parameters.

In a retrospective study comprising 88 patients operated upon with curative intent a number of histopathological parameters of possible prognostic value regarding local recurrence and long-time survival were analysed. Local recurrence within 5 years was found in 28 patients (32%) of which 17 (61%) were diagnosed within the first 2 years. Crude survival rates at 5 and 10 years were 25% and 15%. According to Laurén's classification the results indicated better, but not significant, 5- and 10-year survival for the diffuse type (36% vs 25%). The probability of 10-year survival suggested a better (P = 0.06) prognosis for tumours in the middle third of the stomach, and for patients operated with total gastrectomy (P = 0.025). The probability of recurrence in relation to lymph node involvement suggested a more favourable prognosis (P = 0.06) for patients without lymph node metastases, and in relation to tumour fibrosis a less favourable prognosis for pronounced fibrosis (P = 0.001).

Adipose-tissue concentrations of dioxins and dibenzofurans in potentially exposed patients with malignant-lymphoma or sarcoma.

Epidemiological studies have suggested an association between exposure to phenoxyacetic acids, chlorophenols or their contaminating dioxins (PCDDs) and dibenzofurans (PCDFs) and soft-tissue sarcoma and malignant lymphoma. In this study adipose tissue concentrations of PCDDs and PCDFs were measured in nine patients with sarcoma or malignant lymphoma who had been exposed to phenoxyacetic acids or chlorophenols, or who had other potential exposure to dioxins and dibenzofurans. For comparison concentrations of PCDDs and PCDFs in a group of twelve healthy persons without any known exposure to the above chemicals and living in the same geographical area are presented. Even if no consistent pattern of PCDDs and PCDFs concentrations in the patients was demonstrated, the finding of increased concentrations in all three cases with non-Hodgkin's lymphoma is of special interest. The exposure profiles in relation to measured concentrations of PCDDs and PCDFs are discussed for all patients.

Helicobacter pylori infection: independent risk indicator of gastric adenocarcinoma.

BACKGROUND: Helicobacter pylori has been implicated as a possible etiologic factor in gastric cancer. This case control study was performed to determine the association between H. pylori and gastric cancer, taking into account the possibility of confounding by other background factors. METHODS: Sera were collected from 112 incident case patients with gastric cancer and 103 control patients with nongastroenterological diseases, who were frequency-matched with respect to age and sex. Immunoglobulin G antibodies to H. pylori were identified using the HM-CAP immunoassay (Enteric Products Inc., Wesbury, NY). RESULTS: The prevalence of H. pylori seropositivity was significantly higher (P = 0.002) among case patients than control patients. The odds ratio (OR) was 2.60 (95% confidence interval, 1.35-5.02). The increased OR associated with H. pylori infection was confined to tumors with a noncardia location (OR, 3.06) and men (OR, 4.27). OR increased with decreasing age at cancer diagnosis to reach 9.33 in patients < 60 years of age. Multivariate logistic regression analysis was used as control for potential confounding, but the elevated OR associated with H. pylori infection remained significantly increased. CONCLUSIONS: The results support the hypothesis of H. pylori infection as an independent risk indicator of gastric cancer.

Neutralizing human monoclonal antibodies against Puumala virus, causative agent of nephropathia epidemica: a novel method using antigen-coated magnetic beads for specific B cell isolation.

Human monoclonal antibodies (MAbs) against Puumala (PUU) virus were generated and characterized. Human spleen B lymphocytes were preselected for specific surface immunoglobulin (Ig) using magnetic beads coated with the viral glycoproteins, and subsequently immortalized by Epstein-Barr virus transformation. Four IgG-positive monoclonal lymphoblastoid cell lines (LCLs) were established and have remained stable MAb secretors for over 12 months. Analyses of the antigen and epitope specificities recognized by the MAbs showed overlapping binding patterns of four anti-glycoprotein 2-specific clones. Identical isotypes (IgGl lambda) and isoelectric points (9.2) of the four MAbs suggested that they were derived from the same original clone. The MAbs reacted with eight PUU virus-like strains, but were negative for Hantaan, Seoul, and Prospect Hill viruses in an immunofluorescence assay, indicating binding to a conserved epitope unique for strains associated with the European form of haemorrhagic fever with renal syndrome, nephropathia epidemica. The MAbs neutralized all investigated PUU virus-like strains in a focus reduction neutralization test. The MAb neutralizing activity was significantly enhanced in the presence of human or guinea-pig complement. To stabilize and increase antibody secretion and to reduce the demand for culture medium supplements (e.g. fetal calf serum), three of the monoclonal LCLs were fused with the non-secreting human x mouse partner SPAM-8. Several of the established human x (human x mouse) monoclonal triomas grew faster and produced larger amounts of MAbs when compared with the original LCLs.

Isolation of functionally active intraepithelial lymphocytes and enterocytes from human small and large intestine.

A mild purification method has been developed for the isolation of human intraepithelial lymphocytes (IEL) and enterocytes from the same individual. The isolation procedure includes mechanical disruption of the mucosal layer, treatment with reducing agent and sedimentation followed by Percoll gradient centrifugation. Finally, epithelial cells are removed from the IEL fraction using magnetic beads coated with the anti-epithelial antigen monoclonal antibody (mAb) BerEP4. Leucocytes are removed from the enterocyte fraction using magnetic beads coated with mAbs directed against common leucocyte antigen (CD45). Using this procedure IEL and enterocytes have been isolated from apparently normal jejunal, ileal and colonic tissue specimens. Recoveries of IEL were 7 x 10(5), 4 x 10(5) and 1 x 10(5)/cm2 mucosa from jejunum, ileum and colon respectively. 1-2 x 10(6) enterocytes/cm2 mucosa were recovered from small intestine while the corresponding value for colonic biopsies was approximately 2 x 10(5) enterocytes/cm2. The IEL fraction was pure as judged by the low percentages of B cells, macrophages and BerEP4 positive cells (less than 4%) present in the purified fraction. The enterocyte fraction contained less than 2% CD45+ cells. The two cell fractions were viable and expanded in vitro. Enterocytes expanded spontaneously while IEL required initial stimulation with mitogens. The isolation procedure described here will make it possible to study the function of human IEL, interactions between IEL and enterocytes and the role of both cell types in local immunity.

Ultra-low field MR imaging of hepatic hemangiomas (at 0.02 and 0.04 T).

Twelve patients with cavernous hemangiomas of the liver were studied with computed tomography (CT) and ultra-low field magnetic resonance imaging (MRI). Seven patients were examined at a field strength of 0.02 T and 5 patients at 0.04 T, while 3 patients were studied at both field strengths. On T2-weighted images all hemangiomas had the same characteristic appearance of a homogeneous high signal intensity that has been described at higher field strengths. Signal characteristics of the hemangiomas were the same at 0.02 and 0.04 T, but the higher field strength provided better signal-to-noise (S/N) ratio and hence improved image quality. Homogeneous contrast enhancement was seen in three hemangiomas examined after intravenous administration of gadopentetate dimeglumine. Our results indicate that ultra-low field MRI can be useful in the differential diagnosis of hepatic hemangiomas.

[Information to patients following surgery for cancer]. / Information till patienter efter operation för cancersjukdom.

Patients who are operated upon for cancer tumour are anxious about the postoperative information and the result of the surgical procedure. This investigation was undertaken to study the routines for postoperative information in a department of surgical oncology. Thirty two patients operated for abdominal cancer ("cancer patients") and 36 patients operated for inguinal hernia or gallstone disease ("hernia patients") answered a questionnaire with 16 questions. Sixty nine percent replied. The need for postoperative information about the result of the operation was higher for the cancer patients than for the hernia patients. The cancer patients were given more attention by the doctors postoperatively and were given more opportunities to ask about the operation than the hernia patients. However, there was lack of privacy in the information situation and both groups were informed on the ward in presence of other patients.

The value of routine biochemical tests in discriminating between malignant and benign pancreatic tumours.

The probability that routine hematological laboratory tests of liver and pancreatic function can discriminate between malignant and benign pancreatic tumours, incidentally detected during operation, was investigated. The records of 53 patients with a verified diagnosis of pancreatic carcinoma and 19 patients with chronic pancreatitis were reviewed with regard to preoperative total bilirubin, direct reacting bilirubin, alkaline phosphatase, glutamyltranspeptidase, aminotransferases, lactic dehydrogenase and amylase. Multivariate and discriminant analysis were performed to calculate the predictive value for cancer, using SYSTAT statistical package in a Macintosh II computer. Total and direct reacting bilirubin and glutamyltranspeptidase were significantly higher in patients with pancreatic carcinoma. However, only considerably increased levels of direct reating bilirubin were predictive of pancreatic carcinoma.

Macrophage function and surgery. A clinical review with special reference to phagocytosis.

Present knowledge of macrophage phagocytosis in the context of surgical trauma is reviewed. The historical and morphologic background of the reticuloendothelial system and the mononuclear phagocyte system is surveyed. The physiology of the phagocytic process and methods of measurement are summarized and the influence on phagocytosis of shock, sepsis, cancer, parenteral nutrition and surgical procedures such as liver resection and splenectomy is discussed. Conclusions are that multiple factors may depress macrophage phagocytosis during surgery and that, in order to maintain immune defence balance, traumatic manipulation of tissue and abdominal organs must be minimized.

Fine-needle aspiration biopsy of pancreatic masses.

The results of fine-needle aspiration biopsy of pancreatic masses in 79 patients (percutaneous with ultrasonic guidance in 23 and peroperative in 56) were evaluated and correlated to survival (follow-up at least 2 years). The original biopsy diagnosis was malignancy in 41 patients, histologically confirmed in 19, all but two of whom died of cancer within 18 months. None of the 22 patients without histologic verification of the primary malignant cytodiagnosis survived for 18 months. The fine-needle biopsy showed benign cells in 30 patients, in 13 of whom histologic diagnosis was obtained, revealing carcinoma in seven. Six of these seven died within a year, but of the six with histologically benign lesion, five survived for more than 2 years. All 17 patients without histologic verification of benign aspiration biopsy findings survived more than 24 months. The biopsy diagnosis was inconclusive in eight patients. Four of them proved to have carcinoma and died within 18 months. The sensitivity of fine-needle aspiration biopsy of the pancreas was 76% in this study and the predictive value for malignancy was 100%.

The role of sex and age in yeast cell phagocytosis by monocytes from healthy blood donors.

Possible sex differences and possible age decline of monocyte phagocytosis was studied in healthy blood donors (n = 306). Monocytes from women phagocytosed yeast cells more efficiently than monocytes from men, but the difference was slight. The sex difference was located mainly in the adherence step of phagocytosis. In contrast, no age decline in monocyte phagocytosis could be demonstrated within the age range (18--65 years) represented by healthy blood donors. It is suggested that sex difference and age decline are negligible in the phagocytosis by monocytes from healthy blood donors.

Distribution and scatter of yeast cell phagocytosis by human monocytes in an improved glass surface assay.

The assessment of yeast cell phagocytosis by glass-adherent monocytes was improved by ultrasonication of yeast cells prior to the experiments in order to prevent aggregation, restriction of measurements to completed engulfment regardless of the number of peptides ingested, and stopping of phagocytosis before counting by maintained cooling and the addition of EDTA. The modifications provided a reduced dispersion of individual values, increased engulfment of yeast cells, and decreased numbers of yeast cells only adherent to the monocyte membrane. The improvement made the method more convenient for the study of engulfment by monocytes.

Low field magnetic resonance imaging of focal hepatic masses at 0.02 T.

The diagnostic utility of extremely low field magnetic resonance (MR) imaging was evaluated in 25 patients with focal hepatic masses, including 17 with primary (n = 7) or secondary (n = 10) malignant neoplasms and 8 with benign lesions (6 hemangiomas). The findings were compared with the results of computed tomography (CT). Out of 16 patients with malignant tumors demonstrated by both modalities, the diagnostic information from MR imaging was equal to or better than that from CT in 6 patients and inferior to CT in 10. Shortcomings of MR were mainly due to low signal-to-noise ratio and poor spatial resolution, resulting in an image quality inferior to that obtained at higher field strengths. Considering these facts, together with the long imaging times required, low field MR cannot be recommended for general use in the evaluation of hepatic masses. On the other hand, our results indicate that this technique may be useful in establishing the diagnosis of hepatic hemangioma.

Effect of therapeutic concentrations of anthracyclines on monocyte phagocytosis of yeast cells.

The effect of therapeutic concentrations of doxorubicin, epirubicin, and mitoxanthrone on mature leukocyte function has been examined by measuring phagocytosis of yeast cells by surface-bound monocytes, using a fluorescence-quenching method. There was a 10% inhibition of monocyte phagocytosis by doxorubicin, but epirubicin and mitoxanthrone had no effect on monocyte phagocytosis. Anthracyclines may have a major immunosuppressive effect due to bone marrow depression. The lack of interference with mature monocyte function by epirubicin and mitoxanthrone provides a potential advantage in comparison with the parent compounds.