Significance of lymphocyte fluorescence polarization changes after phytohemagglutinin stimulation in cancer and noncancer conditions.

The double-zone fluorescein fluorescence polarization (FFP) "cancer" test was used to study 540 blood lymphocyte samples from 341 donors, of whom 158 had confirmed cancer: The other donors were noncancer patients, pregnant women, and normal individuals. The FFP response in cancer patients was the reverse of that in normal individuals, but an abnormal response was also obtained in some noncancer conditions, including the chronic inflammatory disorders--rheumatoid arthritis, cholecystitis, and diverticulitis--and in early pregnancy or pregnancy-induced hypertension. Thus the cancer discriminatory value of the test is limited. Examination of its biologic basis suggests that the positive FFP response in cancer and other conditions is due to altered immune status of blood lymphocytes, with associated change in cytoplasmic fluidity affecting the polarization of fluorescence. Incubation of normal blood lymphocytes with cyclic GMP induced an abnormal, cancer-like FFP response.

Zoned immune suppression of lymph nodes draining malignant melanoma: histologic and immunohistologic studies.

Node-to-node heterogeneity of reaction and recognizable patterns of reaction in node groups draining melanoma were sought. Nodes from 72 patients undergoing lymphadenectomy for high-risk, primary melanoma (57) or node-spread melanoma (15) were accurately oriented to the nearest melanoma. Reactivity of paracortex, follicular areas, and sinuses was assessed on a 0-3+ scale. Reactivity was prominent in paracortex and sinuses but varied from node to node within node groups. Nodes nearest to tumor showed least reaction; nodes at intermediate distances from tumor were most reactive, while those farthest away showed mostly little reaction. Variation of nodal reaction that correlated with the node position relative to the nearest melanoma (zoned reaction) was seen in 92% of patients with nodal metastases of melanoma and in 64% of patients with primary malignant melanoma. Follicular and sinusoidal reactions showed no significant zoning. S-100 protein-positive paracortical dendritic cells (PDCs) in tumor-oriented nodes were quantified. PDCs were infrequent in nodes partly replaced by melanoma or located near to melanoma but were numerous in nodes located farther from tumor. Changes of nodal activity (relative stimulation or suppression) correlate with the distance of the node from the nearest deposit of primary or metastatic melanoma.

Immunoglobulins intrinsic to colorectal carcinoma: an unfavorable prognostic association with IgM.

Immunoglobulin (Ig) eluates prepared from cell membrane fractions of 50 colorectal carcinomas were found to contain significantly (P less than .001) less IgA and IgM and marginally less IgG than normal colon mucosa. IgA and IgG were present in all tumors, but IgM was present in only 20 of 50 tumors (40%). Small (less than or equal to the median vol) tumors contained significantly (P less than .05) more IgA and IgG per gram tissue than did large tumors. Tumors with a large (greater than the median) proportion of stroma contained more IgG than did those with less stroma. No associations were found between Ig and amount of viable-appearing tumor tissue, necrosis, mucin production, and staging. Patients whose tumors contained measurable amounts of IgM had a significantly (P = .012) shorter survival than did patients without intrinsic IgM (12 and 27 mo at the 75th percentile, respectively). The difference could not be explained by conventional staging.

Lymphocyte fluorescence polarization changes after phytohemagglutinin stimulation in the diagnosis of colorectal carcinoma.

A lymphocyte fluorescence polarization test that measures reactivity to phytohemagglutinin (PHA) has permitted discrimination of a majority of patients with colorectal carcinoma from noncancer individuals. The test involves separation of two lymphocyte fractions from 20 ml venous blood on a modified leukocyte separation gradient, incubation with PHA for 45 minutes, addition of fluorescein diacetate, and analysis of change in fluorescence polarization. Of 19 colorectal patients tested preoperatively, 13 had a positive stimulation index, 3 a zero index, and 3 a negative index. Of 7 patients with other malignant neoplasms, a positive value was obtained in 6 and a negative value in 1. Of 14 patients with other diseases, a negative value was obtained in 9, zero in 3, and positive in 2. Of 31 normal donors a negative value was obtained in 27, zero in 2, and weak positive in 2. This rapid test promises to develop into a useful cancer-diagnostic method, and elucidation of its biological basis should identify a new cancer marker.

Immunomorphological features of prognostic significance in Dukes' Class B colorectal carcinoma.

Histological slides of primary tumors and regional lymph nodes from 134 unselected patients operated on for colorectal carcinoma of Dukes' Class B were assessed semiquantitatively for the presence of perivascular lymphocyte cuffing in the muscular layers and pericolic/subserosal fat immediately subjacent to the tumors and for paracortical hyperplasia in the regional lymph nodes. These two immunomorphological features related significantly to each other (p less than 0.05), and their combined presence related signifcantly to favorable disease-free interval (p = 0.02) and to survival (p = 0.04), making possible the identification of a subgroup of approximately one-third of Dukes' B class patients with an estimated better than 85% chance for 5-year recurrence-free survival.

Correlation of regional lymph node in vitro antitumor immunoreactivity histology with colorectal carcinoma.

Lymph nodes from resected specimens of human colorectal carcinoma were investigated for in vitro lymphocyte cytotoxicity against primary cultures of autologous tumor cells. Regional lymph node lymphocytes were cytotoxic in 32 of 142 cases (23%). Altogether 200 nodes were examined and the cytotoxicity correlated directly with sinus histiocytosis, seen in 43 nodes from 35 cases, and with hyperplasia of B- and T-lymphocyte areas combined, seen in 92 nodes from 65 cases. Lymph nodes with combined B- and T-cell hyperplasia were significantly more common in cases of good tumor differentiation. The findings suggest that sinus histiocytosis and hyperplasia of both major lymphocyte populations are morphological expressions of in vitro antitumor immunoreactivity in the regional lymph node.

High-sensitive C-reactive protein level and oxidative stress-related status in former athletes in relation to traditional cardiovascular risk factors.

OBJECTIVE: To analyze systemic and cellular oxidative stress-related indices as well as C-reactive protein level in former top-level athletes in relation to traditional cardiovascular risk factors. METHODS: A cross-sectional study was performed in 53 former male athletes and 25 sedentary controls (age range: 39-59 years). We measured anthropometric factors (BMI, fat percentage, WHR), resting blood pressure (SBP, DBP), serum cholesterol (CHOL), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), triglycerides (TG), total antioxidant status (TAS), oxidized LDL-C (oxLDL), diene conjugates (DC), glutathione redox status, high-sensitive C-reactive protein (hsCRP), and leisure-time physical activity. RESULTS: Physically active former athletes had significantly lower mean overweight (BMI, fat percentage, WHR), better spectrum of atherogenesis indicators (CHOL, HDL-C, TG, TG:HDL-C ratio) and lower oxidative stress (oxLDL, oxLDL:LDL-C ratio, DC) values than sedentary ex-athletes. No significant differences in these variables were found between the sedentary ex-athletes and control group. Significant associations were found between physical activity (METs), SBP, DBP, hypertension, CHOL, HDL-C, TG, TG:HDL-C ratio, oxLDL, oxLDL:LDL-C ratio, DC and hsCRP. CONCLUSIONS: A physically active lifestyle is related to a lower cardiovascular disease (CVD) risk profile including a substantially lower systemic and cellular oxidative stress status as well as C-reactive protein level in middle-aged men.

T helper lymphocyte depression in early human pregnancy.

Peripheral blood lymphocyte subclasses were determined in 60 women with normal pregnancies, 20 from each trimester, and in 20 controls using automated flow cytofluorimetry. The cells were stained with the monoclonal antibodies OKT3, OKT4 and OKT8 to stain total T cells, T helper and T suppressor-cytotoxic lymphocytes, respectively. A polyvalent rabbit anti-human Ig serum was used to stain B lymphocytes. Absolute numbers of T lymphocytes were significantly reduced in both the first and second trimesters. This was due to a significant decrease in T helper lymphocytes and a smaller, statistically not significant, reduction in the number of T suppressor lymphocytes. There was no significant change in lymphocyte subclasses during the third trimester. Total lymphocyte numbers were normal throughout pregnancy.

Changes in body fluids during endurance rowing training.

The aim of this study was to investigate the influence of long-lasting, extensive rowing training on body fluids in 12 experienced rowers (23.3 +/- 6.3 years, 188.7 +/- 5.9 cm, 82.0 +/- 10.8 kg, body mass index [BMI] 23.7 +/- 2.1). All rowers had a light traditional breakfast about 3 h before the training. The subjects rowed 2 h 17 min and covered 22.6 +/- 2.5 km. Before and immediately after the training session and after 30, 60, and 120 min of recovery, body fluid balance was measured using a multiple-frequency impedance device (Multiscan 5000, Bodystat Ltd., UK). Body resistance was measured at the right side of the body. Extracellular (ECW), intracellular (ICW), and total (TBW) body water were measured at 5, 200, and 50 kHz, respectively. Blood hemoglobin (Hgb) and hemotocrit (Hct) were measured, and changes in blood volume (BV), plasma volume (PV), and cell volume (CV) were measured according to Dill and Costill (1974). The body weight of the rowers decreased significantly (p < 0.05) from 82.0 +/- 10.8 kg to 80.6 +/- 11.2 kg during the endurance training. There were no significant changes in TBW immediately after the training session. Significant changes in the content of TBW occurred during the first 30 min of recovery (48.7 +/- 4.4 to 47.9 +/- 4.1; p < 0.05). The amount of ECW also decreased significantly during the first 30 min of recovery (23.3 +/- 2.0 to 22.7 +/- 1.91; p < 0.05). There were no significant changes in the balance of ECW. BV decreased during the training session by 2.5 +/- 5.2% (p < 0.05). There were significant relationships (p < 0.05) between the covered distance and the following parameters after the training session: body weight (r = -0.75), TBW (r = -0.75), and ECW (r = -0.83). Changes in the blood parameters did not correlate significantly with changes in body fluids and training volume. It was concluded that extensive rowing training significantly changed the balance of body fluids in rowers.

Immunohistological patterns of carcinoembryonic antigen in colorectal carcinoma. Correlation with staging and blood levels.

Forty-four primary adenocarcinomas of the large bowel and 2 liver metastases were stained for carcinoembryonic antigen (CEA) in tissue sections by indirect immunofluorescence. All tumours were positive and showed either one or more of 3 different patterns--luminal; linear at surface of the tumour cells; cytoplasmic. In most cases (83%), two or all 3 patterns were seen in the same or in different parts of a tumour. The immunohistological staining was concordant with preoperative blood levels of CEA in 31 cases (67%) in that 26 tumours showed strong immunofluorescence associated with blood CEA above 2.5 micrograms/l, and 5 showed weak staining and blood CEA values less than 2.5 micrograms/l. However, in 7 strong staining was associated with low blood CEA, and in 8 weak staining was associated with high blood levels. The dissociation between histological and blood CEA findings in 1/3 of the cases, together with the marked variation within the same tumour and differences between one of the primaries and its liver recurrence, suggest that CEA immunohistology is of no better prognostic value than blood CEA levels. There was no association between CEA immunohistology and tumour staging or differentiation. However, blood CEA levels were significantly higher in tumours with extensive local or distant spread (stage D) and in poorly differentiated tumours.

Mucinous colorectal carcinoma: immunopathology and prognosis.

A total of 519 colorectal carcinomas were examined for the presence or absence of mucinous differentiation by means of microscopical morphometry. Of these, 28% had objectively measurable amounts of mucinous tumour epithelium. Tumours with > 50% mucinous areas (14%) had significantly poorer prognosis than non-mucinous in stages A and C, while mucinous differentiation did not correlate with prognosis in stages B and D. Lymph nodes regional to mucinous tumours had significantly less paracortical response, and those with < 50% mucinous differentiation, significantly less perivascular lymphocyte cuffing at the tumour margins. These lymph node and stromal compartments are putative T-lymphocyte areas, and hence our findings suggest that mucinous tumours are either less stimulatory or perhaps inhibitory of cell-mediated immunity.

Regional lymph node and stromal immunomorphology in colorectal carcinoma and relation to tumour spread.

A quantitative morphometric study of lymphocyte patterns in the stroma and regional lymph nodes was made in 509 cases of colorectal adenocarcinoma and 17 non-invasive adenomas. An increase in the perivascular lymphocytes, and in the size of lymph nodes, germinal centres and paracortical areas was most obvious in 'localized' invasive tumours and significantly more in Dukes' stage B than in stage C. Stage C1, defined as cases where the primary tumours were confined to the wall but with lymph node spread, showed hardly any perivascular lymphocyte aggregates, although in the regional lymph nodes there was relative paracortical, i.e. T-lymphocyte, hyperplasia. It is concluded that the presence of cuffs of perivascular lymphocytes at the tumour edge, and the increased size of tumour-free regional lymph nodes together with the relative and absolute abundance of germinal centres (B-lymphocyte) and paracortical areas (T-lymphocyte), are stage dependent. This is most obvious in stage B. These morphological expressions of host immunoreactivity may well reflect favourable anti-tumour mechanisms.

Persistence of organ- and iso-antigens in vitro in a long-term culture cell line of colonic carcinoma.

Cells of colonic carcinoma line HT-29, cultured over more than 170 generations and extensively used in immunological studies of patients with colorectal tumours, still express several immunologically valuable characteristics presumably present since its inception. The cell line can induce a poorly differentiated adenocarcinoma in the nude mouse, it has human antigenic characteristics, it expresses blood group A antigen, and produces a colon-specific mucin and CEA, though not colon cancer-specific mucin (CCM). It remains useful as a target for in vitro testing of anti-tumour immunoreactivity in colorectal cancer patients.

Germinal centre hyperplasia: an unfavourable immune response in recurrent malignant melanoma.

A quantitative immunomorphological analysis of actively responding B-(germinal centre) and T-lymphocyte (paracortical) areas in 187 patients with malignant melanoma recurrent in the regional lymph nodes has been made. The results show that tumour involved nodes have significantly increased germinal centre and decreased paracortical response in comparison with uninvolved lymph nodes. Patients treated with surgery alone whose uninvolved nodes had hyperplastic (defined as exceeding the median) germinal centre areas, had significantly shorter survival than those without germinal centre hyperplasia. No association was found between lymph node immunomorphology and survival in BCG treated patients.

Relationship between subcutaneous fatness and leptin in male athletes.

PURPOSE: Circulating leptin is low in trained subjects and closely related to body fat content. However, data are scarce as to whether differences exist in the relationship between different estimates of adiposity, metabolic parameters, and leptin in endurance- and resistance-trained male athletes. We investigated this relationship with special emphasis on subcutaneous fatness and its distribution. METHODS: 20 endurance (ET) and 17 resistance (RT) athletes recruited from different kind of sports were studied. Fat-free mass (FFM) was estimated by means of impedance and fat mass (FM) was calculated. Subcutaneous fat (SAT) and its distribution was measured by means of the optical device Lipometer at 15 body sites (SAT-layers; from 1-neck to 15-calf) on the right side of the body. Fifteen SAT-layers were summed to calculate SAT. Blood samples were obtained for determination for leptin, insulin, and glucose. Insulin resistance was calculated through the fasting insulin resistance index (FIRI; [insulin x glucose/25]). RESULTS: RT-athletes had a greater body mass and body fat content than ET-athletes, but no differences were found for leptin and metabolic parameters. In all athletes, estimates of adiposity were correlated to leptin. However, in ET-athletes FM (P < 0.05), FFM (P < 0.05), and SAT (P < 0.001) but not metabolic parameters were correlated to leptin. In RT-athletes, SAT (P < 0.0001), metabolic parameters (all P < 0.05), but not FM and FFM were in significant relationship with leptin. Stepwise regression revealed SAT as the main determinant for the variation in leptin in all athletes (adj. R(2 )= 0.52, P < 0.0001). CONCLUSION: The results suggest that estimates of adiposity and metabolic parameters are associated with leptin in a sport-specific manner. Whereas leptin might be regulated by overall subcutaneous fatness in athletes, our study does not imply a main influence of fat patterning on leptin in this group of trained subjects.

Physical activity, musculoskeletal disorders and cardiovascular risk factors in male physical education teachers.

BACKGROUND: To compare the occupational and leisure-time physical activity (LTPA) patterns, musculoskeletal disorders and cardiovascular disease risk factors among the physical education teachers (PETs) and their fellow workers (controls). DESIGN: a case-control study. PARTICIPANTS: 86 male PETs and 102 other teachers aged 51-72 years old. MEASURES: the questionnaire was applied to determine the occupational activity and LTPA level, musculoskeletal disorders, general health status and lifestyle patterns. Anthropometric parameters, resting blood pressure values, and physical working capacity were measured. Concentrations of total cholesterol (CHOL), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol, triglycerides and glucose were determined and the HDL-C/CHOL ratio was calculated. RESULTS: PETs had maintained their physically active lifestyle - 59.3% of them were regularly active in their leisure time. The results of the questionnaire did not reveal any major between-group differences in their health status and health habits (smoking, alcohol consumption, etc.). PETs had a significantly lower adjusted risk of overweight (OR 0.63, 95% CI, 0.21-0.94), hypertension (OR 0.25, 95% CI, 0.13-0.47) and all musculoskeletal disorders (during the past year) (OR 0.52, 95% CI, 0.29-0.94) in comparison with the control group. CONCLUSIONS: It is reasonable to assume from our study that long-term physical activity is associated with a lower risk of overweight, hypertension and musculoskeletal disorders in PETs in comparison with sedentary fellow workers.

Blood lactate recovery and perceived readiness to start a new run in middle-distance runners during interval training.

The aim of this study was to establish to what extent ratings of perceived readiness to start a new run are dependent on recovery of blood lactate and heart rate during passive recovery between interval runs (4 x 2000 m) of increasing intensity by 15 male college-level middle-distance runners. The Perceived Readiness Rating scale was administered at each minute of recovery. Blood lactate concentration was measured immediately after runs at Min. 3 of recovery after the first and second runs, and, in addition, at Min. 6 of recovery after the third and fourth runs. Heart rate was recorded continuously during the runs and during recovery. The obtained correlation between blood lactate recovery and perceived readiness ratings after the third and fourth runs were -.36 and -.56, respectively (p<.05). Consequently, blood lactate concentration recovery appears to influence the middle-distance runners' perceived readiness estimation to begin a new run. The Perceived Readiness Rating applied by us appears useful in the training of middle-distance runners.