Mechanical control of cyclic AMP signalling and gene transcription through integrins.

This study was carried out to discriminate between two alternative hypotheses as to how cells sense mechanical forces and transduce them into changes in gene transcription. Do cells sense mechanical signals through generalized membrane distortion or through specific transmembrane receptors, such as integrins? Here we show that mechanical stresses applied to the cell surface alter the cyclic AMP signalling cascade and downstream gene transcription by modulating local release of signals generated by activated integrin receptors in a G-protein-dependent manner, whereas distortion of integrins in the absence of receptor occupancy has no effect.

A morphometrical analysis of lymph node responses to tumors of different immunogenicity.

An immunogenic tumor and a nonimmunogenic tumor were used in C57BL/Ka and CBA/Rij mice, respectively, in a study in which lymph node morphology was quantitatively measured. Lymph nodes were cut in semiseries, and a morphometrical analysis was done on the paracortical and cortical areas as major parts of the active lymph nodes. These parameters could be expressed with confidence in both absolute and relative numbers, since a strong correlation existed between the weight of lymph nodes and the summated surface areas. To determine whether the regional lymph node plays a major role in this response, we performed similar analyses of two nonregional lymph nodes. It was observed that immunogenicity, defined as the ability to evoke an immune response that significantly influences tumor cell take and tumor cell growth, was not relevant in evoking a morphologically evident response. In absolute numbers, the response evoked was strongest in the regional lymph nodes in both systems. However, the percentages of paracortical and cortical areas in the lymph nodes reacting were not essentially different from one node to another during the growth of both tumors. This observation was confirmed by an analysis of variance. These findings suggest that great caution must be exercised when one is interpreting morphological lymph node changes as evidence of an influence of immunological reactions on tumor cell growth.

Alteration of retinol-binding-protein concentrations by the synthetic retinoid fenretinide in healthy human subjects.

Normal subjects received fenretinide (HPR), 200 mg/d, on three schedules. Schedule 1 was treatment for 28 d. Schedule 2 consisted of 14 d of treatment, 3 d hiatus, and a second drug course of 14 d, 10,000 IU vitamin A was administered during the 3-d hiatus. Schedule 3 was 14 d of treatment followed by a rest period of 7 d and then 14 d of treatment. Increase in plasma HPR was accompanied by an even higher increase in the metabolite N-(4-methoxyphenyl)-all-trans-retinamide (MPR). The administration of HPR was associated with a significant reduction in retinol-binding protein (RBP), which returned to pretreatment values after the drug treatment was discontinued. Reduction of plasma retinol was also observed. Use of interrupted schedules with resting periods of 3 and 7 d changed HPR, MPR, and RBP concentrations in plasma. Addition of vitamin A did not affect the pattern of the measured variables in the plasma.

Circulating IgA-immune complexes in Henoch-Schönlein purpura. A longitudinal study of their relationship to disease activity and vascular deposition of IgA.

In Henoch-Schönlein purpura immune complexes in inflamed vessel walls characteristically contain immunoglobulin A(IgA). To determine whether IgA is also the predominant immunoglobulin in circulating immune complexes, we compared the results of three immune complex assays with specificities for different classes of immunoglobulins in a longitudinal study of 37 patients (30 children and seven adults) with Henoch-Schönlein purpura. Circulating IgA-containing immune complexes were detected by their reactivity with a low avidity anti-IgA antibody in 27 of the 37 patients. IgA was simultaneously present in cutaneous vessel walls in 95 percent of the patients with circulating IgA-containing immune complexes. High levels of IgA-containing immune complexes were found only during the initial phase of the disease. Immune complexes containing bound complement breakdown products were demonstrated by binding to conglutinin. IgA was found in these immune complexes in 17 patients, IgG in 17 and IgM in nine patients. There was no apparent relation with the class of immunoglobulin in the deposits. Conglutinin-binding immune complexes were present later in the course of the disease and after remission. C1q-binding immune complexes were only found in two patients. These findings suggest that immune complexes-containing IgA may initiate the vasculitis of Henoch-Schönlein purpura, whereas complement-reacted immune complexes containing immunoglobulins of the other classes appear in the circulation in a later phase.

Mononuclear cells in renal allografts. Correlation with peripheral blood T lymphocyte subpopulations and graft prognosis.

Cellular infiltrates in 20 renal allograft biopsies taken at the time of acute rejection episodes were analyzed with antibodies reactive with monocytes (BRL antihuman monocyte monoclonal antibody), T lymphocyte subpopulations (OKT- and Leu-series of monoclonal antibodies), and B lymphocytes (heterologous antihuman IgM antibodies). For demonstration of the various mononuclear cells, an indirect immunoperoxidase technique was used. The number of monocytes in the infiltrates varied from 10-20%; the number of B lymphocytes was always below 10%. The T lymphocytes accounted for 50-90% of the total number of mononuclear cells. The OKT4/OKT8 ratios for the T lymphocytes in the graft infiltrates were correlated with the peripheral blood OKT4/OKT8 ratios measured by indirect fluorescence and flow cytometry. The OKT4/OKT8 ratios for perivascular infiltrates correlated far better with peripheral blood OKT4/OKT8 ratios (r = 0.72) than did the OKT4/OKT8 ratios for interstitial infiltrates (r = 0.58). Low or inverted OKT4/OKT8 ratios and low or inverted Leu 3a/Leu 2a ratios were associated with a high risk of irreversible graft rejection (P less than 0.001 for perivascular infiltrates).

Ultrastructural localization of the low molecular weight protease inhibitor in human bronchial glands.

Light microscopically the low molecular weight protease inhibitor (LMI) and lysozyme have both been demonstrated in the cytoplasm of serous cells of bronchial glands. By immunoelectron microscopy LMI was demonstrated in secretory granules of these serous cells. Many granules showed a peripheral or bean-shaped staining pattern, other granules were uniformly stained or not stained at all. In contrast to this latter finding, virtually all granules were found positive for the presence of lysozyme, suggesting that there is no association of LMI with lysozyme at the ultrastructural level.

Adult T-cell leukemia: a report on two white patients.

Two white European males are reported with adult T-cell leukemia (ATL), a disease first described in Japan, but recently also in the U.K. and U.S.A. Both patients presented with lymphadenopathy, but without a mediastinal mass. In addition, one patient had skin infiltrates and the other had hepatosplenomegaly. Morphologic and ultrastructural examination of the blasts in bone marrow and lymph node biopsy revealed a predominance of polymorphic lymphoid cells with pronounced nuclear irregularities and a semi-mature chromatine pattern. Histopathology of the lymph nodes showed a diffuse infiltration with medium-sized lymphoblasts with irregular nuclei. The blasts in the bone marrow formed E rosettes with sheep erythrocytes, lacked terminal deoxynucleotidyl transferase (Tdt) activity but expressed the Ia-like antigen; although the majority of the cells reacted with a polyclonal anti-T-cell serum, they were negative for OKT3. In one patient a helper/inducer phenotype (OKT4+) was found in the lymphoblasts of bone marrow and lymph node, while in the other only in the lymph node. The difference between bone marrow and lymph node phenotype is discussed. To our knowledge, these are the first two European patients reported with ATL, a disease clearly different from convoluted T-cell acute lymphocytic leukemia.

Combination therapy of pentoxifylline and TNFalpha monoclonal antibody in dextran sulphate-induced mouse colitis.

BACKGROUND: Tumour necrosis factor-alpha (TNFalpha) has been suspected of playing an important role in the pathogenesis of inflammatory bowel diseases, and has become a target for the treatment of these diseases. Open-label, placebo controlled studies have shown that engineered CDP571 and chimeric anti-TNF antibody (cA2) provide a significant benefit in Crohn's disease. Since these antibodies have to be used repeatedly to maintain remission in inflammatory bowel disease, there is a concern that their use may compromise host defence and produce toxic side-effects. METHODS: We evaluated the combined use of mouse specific TNFalpha mab (25 microg/mouse, Endogen) and pentoxifylline (PF, 100 mg/kg/day, p.o., TNFalpha release inhibitor) in the DSS (3% dextran sulphate solution) model of mouse colitis. Colitis was induced by the feeding of 3% DSS for three cycles. The study groups were: Group I: single injection of rat anti-mouse IgG, Group II: single injection of TNFalpha mab, Group III: daily PF for three cycles, Group IV: single injection of TNFalpha mab + PF for three cycles, Group V: TNFalpha mab at the beginning of each cycle (three injections) and Group VI: TNFalpha mab (three injections) + daily PF for three cycles. Daily disease activity (DAI) was measured throughout the study. At the end of each cycle, colon tissue was processed for histology, myeloperoxidase (MPO) and plasma TNFalpha. RESULTS: Mice treated with a single injection of TNFalpha alone or TNFalpha mab + PF showed significantly lower DAI, inflammation scores and ulcer index compared with the IgG treated group. Mice treated with TNFalpha mab + PF had no ulcers. Multiple injections of TNFalpha mab or TNFalpha mab + PF showed greater inhibition in DAI and cytokines in the first two cycles. However, in the third cycle, multiple injections of TNFalpha mab showed adverse proinflammatory effects. CONCLUSION: The simultaneous administration of pentoxifylline and TNFalpha mab may enhance therapeutic outcomes in inflammatory bowel disease and reduce the side-effects associated with the repeated use of TNFalpha mab.

Prospective evaluation of prognostic value of morphometry in patients with primary breast cancer.

In a prospective study the predictive value of a multivariate morphometric prognostic index was evaluated in 195 patients with primary breast cancer who had not been treated with any form of chemotherapy or hormonal treatment. The presence or absence of distant tumour recurrence combined with the scores of the prognostic index were compared with the survival curves predicted in a previous study. The value of the presence of lymph node metastases, number of positive nodes, tumour size, mitotic activity index, and oestrogen receptor status in prediction of prognosis were also investigated. In agreement with the results of the previous retrospective study, the prospective use of the index had the strongest predictive prognostic value, followed by the mitotic activity index. Statistical analysis showed that the actual prognoses of 43 of the 195 patients (22%) were more accurately determined by the prognostic index rather than by using the presence of the lymph node metastases as the classifying variable. The prognostic index is consistently reproducible by different technicians; it is a reliable method of predicting distant recurrence of tumour and hence the prognosis of patients with primary breast carcinoma. It provides more prognostic information than the presence of lymph node metastases alone, and the index should be incorporated in routine pathology reports.

Morphometry and breast cancer. II. Characterisation of breast cancer cells with high malignant potential in patients with spread to lymph nodes: preliminary results.

The prognostic value of clinical, quantitative, and qualitative microscopical features of both the primary tumour and also of the affected lymph nodes were investigated in 71 patients with breast cancer with spread to lymph nodes (T X N + M0). Age, tumour size, and localisation of the tumour comprised the clinical features; morphometry included assessment of the cellularity index, the mitotic activity index, and seven nuclear indices; the qualitative features investigated were histological type and grade, nuclear grade, oestrogen receptor content, number of lymph nodes affected, capsule infiltration of the nodes, presence of metastatic deposits in the efferent lymph vessels, percentage area of lymph node occupied by tumour. Immunohistochemistry was performed to show the presence of carcinoembryonic antigen and peanut agglutinin. All the patients had a minimum follow up of 24 months (maximum 48 months, mean 36 months). Analysis of the results showed that the combined results of morphometry (of the primary tumour and the axillary lymph node metastatic deposits) yielded more information than analysis of axillary lymph node state, or morphometry of the primary tumour, or the lymph node metastases alone. Patients with a nuclear axes ratio of greater than 1.41 in the primary tumour and greater than 1.36 in the lymph node metastatic deposits were less likely to develop distant metastases than patients with values below any of these thresholds (recurrence rates 5.2% and 46%, respectively). Thus the preliminary results of this prospective study indicate that morphometry provides important prognostic information in patients with breast cancer that has spread to lymph nodes.

Increased numbers of IgE containing cells in gastric and duodenal biopsies. An expression of food allergy secondary to chronic inflammation?

Gastric and duodenal biopsies from 2543 patients with abdominal complaints were sent to the Department of Pathology of the Stichting Samenwerking Delftse Ziekenhuizen in 1980 and 1981 and screened for plasma cells containing IgE using an indirect immunoperoxidase technique. Increased numbers of IgE containing cells were found in 2.6% of the patients. These patients all suffered from a variety of chronic non-specific inflammatory disorders of the upper gastrointestinal tract. No specific clinical, endoscopical, or histological picture could be found. The results suggest that increased numbers of plasma cells containing IgE in biopsies from the upper gastrointestinal tract are an expression of IgE mediated type I allergy presumably to food constituents as a secondary complication of chronic non-specific gastric and duodenal inflammation in these patients.

Monitoring serum CEA in women with primary breast tumours positive for oestrogen receptor and with spread to lymph nodes.

Serum carcinoembryonic antigen concentrations (serum CEA) in 80 patients with primary breast cancer were measured preoperatively, one month after operation, and thereafter serially every third month. These data were related to histological and morphometric features of the primary breast carcinoma and the lymph node metastases and to clinical follow up data. Analysis of the serum CEA values showed significant correlations with size of tumour, the presence of lymph node metastases, oestrogen receptor, and occurrence of distant metastases. Furthermore, the results indicated that serial determination of serum CEA in the first two years after operation may be useful in monitoring for the occurrence of distant metastases in patients with metastatic spread to lymph nodes and with large (greater than or equal to 2 cm) primary breast tumours positive for oestrogen receptor. In agreement with other studies, however, it was found that the predictive value of serum CEA concentrations in general is weak and costs may prohibit the implementation of the routine assessment of CEA concentrations.

Use of statistical evaluation of antigen profiles in differential diagnosis between colonic and ovarian adenocarcinomas.

A study was carried out to determine whether it was possible to classify colonic and ovarian adenocarcinomas by their antigen profile. Colonic and ovarian adenocarcinomas were immunostained with a panel of antibodies which have a limited specificity for colon (parlam-4, 19.9, anti-secretory component) and ovary (OV-TL3 and OC125) and the most discriminatory antibodies were selected by stepwise linear discriminant analysis. For frozen material OV-TL3 and OC125 were the best classifying antibodies. Although OC125 had better discriminative power, for paraffin wax embedded material parlam-4 was selected as the best classifying antibody. OC125 had no additional effect on the classification of a tumour. These antibodies were subsequently tested on an independent test set of primary and metastatic adenocarcinomas of colonic and ovarian origin. When ovarian posterior probabilities of less than 0.1 and greater than 0.9 were selected as cut off points for a positive identification of colonic or ovarian origin (jackknifed classification method), no adenocarcinoma was incorrectly identified as ovarian carcinoma in frozen material. The same trend was noticed for paraffin wax embedded material. Statistical analysis of antigen profiles can be helpful in defining the colonic or ovarian origin of an adenocarcinoma when routine microscopy does not yield a definitive result.

Retrospective study of prognostic importance of DNA flow cytometry of urinary bladder carcinoma.

Cellular DNA content was determined by flow cytometry on routinely processed paraffin sections of 61 primary and untreated transitional cell carcinomas of the urinary bladder, and correlated with tumour grade and stage and clinical follow up. All 16 (25%) grade 1 carcinomas were diploid and all 11 (20%) grade 3 tumours were aneuploid. The 34 (55%) grade 2 carcinomas comprised 13 (40%) diploid and 21 (60%) aneuploid cases. Among the 37 superficial carcinomas (stage Ta and T1), 25 (65%) were diploid; 20 (85%) of the 24 advanced tumours (stage T2 to T4) had aneuploid tracings. Ploidy was a significant prognostic indicator (p: 0.006) of five year survival. The initial presence of aneuploidy in superficial bladder carcinoma (stage Ta and T1) is a strong argument for more aggressive treatment than is customary.

Quantitative morphometry of squamous cell hyperplasia of the larynx.

The histopathological diagnosis of squamous cell hyperplasia of the larynx is very subjective. Since morphometry is highly reproducible, this method was applied to routine processed slides of 45 such lesions to assess objectively the epithelial characteristics. In each case measurements of nuclei of 50 cells in the basal, intermediate, and superficial cell layers were carried out. The data were analysed statistically. The findings suggest that quantitative morphometry may be helpful for the histopathological classification of squamous cell hyperplasia of the laryngeal mucosa.

Improved gas chromatographic analysis of chlorpromazine in blood serum.

Reliable determinations of chlorpromazine levels in blood serum samples obtained from patients were accomplished by electron capture gas chromatography. By using modifications of the procedure to insure stability of the sample, minimal losses during sample preparation and gas chromatography, and by selecting appropriate operating parameters of the electron capture detector, excellent agreement was obtained in replicate analyses with a limit of sensitivity of 1 ng/ml in 1 ml of plasma.

The effect of selenium on phagocytosis in humans.

The effect of selenium on immune responses in animals and humans is controversial. It has been reported that phagocytosis as a part of the immune function is affected by selenium deficiency. We conducted a study to investigate the effect of selenium on the phagocytic function of polymorphonuclear leukocytes (PMNs) in normal healthy individuals before and after selenium supplementation. Ingestion of sodium selenite 400 micrograms/day (182.8 micrograms pure selenium) resulted in a significant increase in plasma selenium levels. The phagocytic function of PMNs was measured by ingestion of Oil Red O paraffin droplets and chemiluminescence tests. The phagocytic function was increased, but the results before and after selenium supplementation were not significant. It was concluded that inorganic selenium was not an efficient stimulating agent of phagocytosis in humans.

MRI of normal or cancerous laryngeal cartilages: histopathologic correlation.

MRI appearances of laryngeal cartilages, normal or invaded by cancer, are still relatively unfamiliar to most clinicians. Twelve primary laryngeal tumors out of a series of 65 patients which have been investigated by MRI were examined postoperatively by macroscopic and microscopic sectioning of the surgical specimens. Images were obtained with a 0.6 Tesla superconductive system using a solenoid surface coil. The authors emphasize the value of a combined use of T1-weighted and balanced (relatively T2-weighted images with still T1-characteristics) Spin Echo images. T1-weighted images permit differentiation between pathological and normal bone marrow. Balanced images allow separation between nonossified cartilage and tumor tissue. MRI is an additional tool in the diagnostic workup of cartilage invasion by tumor.

Magnetic resonance imaging vs palpation of cervical lymph node metastasis.

In a series of 100 patients with head and neck carcinoma, the preoperative histopathologic findings of palpation and magnetic resonance imaging were compared with regard to both laterality and lymph node level (I through V). The overall error for palpation in detecting affected sides was 32%. Gadolinium-enhanced magnetic resonance images reliably upgraded 60% of the clinically negative necks, the overall error of magnetic resonance imaging being 16%. However, for both modalities, the sensitivity per level was too low to allow for selective neck dissections in case of only one positive level. These findings show that apart from primary tumor grading, magnetic resonance imaging can improve the preoperative grading of cervical lymph nodes. In selected cases, this may change the treatment plan to a "wait-and-see" policy or a more conservative type of neck dissection.

Human papillomavirus DNA and genotypes: prognostic factors for progression of cervical intraepithelial neoplasia.

A retrospective study of 227 patients presenting with abnormal cervical cytology was conducted to investigate the relationship between human papillomavirus (HPV) and progression of untreated cervical intraepithelial neoplasia (CIN) lesions. All patients had colposcopically directed biopsies for histologic diagnosis. The patients were followed cytologically and colposcopically for a mean of 19 months (range 6-42 months). Progression of a cervical lesion was defined as progression to a higher CIN grade confirmed histologically by directed biopsy. HPV DNA detection was done on material remaining from the cervical swabs by the general primer polymerase chain reaction (PCR) and type-specific PCR method, which made the detection of HPV types 6, 11, 16, 18, 31, 33 and not yet sequenced DNA types (X) possible. The presence of HPV DNA increased with the severity of the lesion (P < 0.001). In CIN III, a 100% HPV DNA prevalence was found, with HPV type 16 being the most prevalent type in 75%. Progression was significantly related to the presence of HPV DNA, in particular HPV type 16. The percentage of progressive disease was 21% in the case of HPV DNA positive lesions (n = 130) and 29% in the presence of HPV type 16, whereas HPV DNA negative lesions (n = 97) showed no progression. The detection of HPV DNA and HPV genotype can be used to identify patients with high-risk cervical lesions, since the presence of HPV DNA and genotype 16 in particular are closely related to CIN progression.