Cytogenetic analysis of peripheral blood lymphocytes of workers occupationally exposed to styrene

The genetic risk run by workers occupationally exposed to styrene vapors was assessed in two different plants A and B, using the cytogenetic analysis of peripheral blood lymphocytes. In plant A engaged in the manufacture of polystyrene vessels the mean styrene exposure level was found to range between 70 and 150 mg . m-3, in plant B manufacturing sports boats, plastic slides for children and plastic guard-stones it reached the level of about 200 mg . m-3. The rate of aberrant cells (AB.C.) found in plant A workers (N = 36) at the time of first sampling was 1.38% and the value of break/cell (B/C) ratio was 0.015; at the second sampling the rate of AB.C. was 1.41% and the B/C ratio was 0.014. The group of matched controls (N = 19) was found to have 1.26% of AB.C. and 0.014 breaks per cell. Plant B workers (N = 22) exhibited at the first sampling 1.72% of AB.C. and their value of B/C ratio was 0.018, the group of matched controls (N = 22) had 1.36% of AB.C and the B/C ratio 0.015; the respective values at the time of second sampling were 2.81% for AB.C. rate and 0.029 for B/C ratio in the exposed and 1.89% for AB.C. rate and 0.021 for B/C ratio in the control group. It is concluded that styrene exposure levels below 100 mg . m-3 do not pose any serious genetic risk for the exposed population groups. The variations found in the degree of chromosome injury by smoking habits, drug intake pattern, or sex were not statistically significant.     

Chromosomal abnormalities in soft coal open-cast mining workers

Chromosome aberration tests on cultured peripheral blood lymphocytes were carried out in two groups of open-cast mining workers engaged in stripping operations (Group I) and digging operations (Group II), in a group of coal cleaning plant workers used as a positive control, and in a group of nonexposed subjects serving as matching controls. Peripheral blood samples in group I and group II miners were collected in March and June of 1984, the antimutagenic potential of long-term prophylactic vitamin administrations (1 g of ascorbic acid given daily for a period of 3 months and 100 mg of vitamin A a day for two 5-day cycles) initiated after the first blood sampling was tested in group II miners. The frequency of aberrant cells (% AB.C.) found in the cultured lymphocytes of group I miners was increased at both times of measurement (2.41% AB.C. and 2.29% AB.C.), in group II miners given the vitamin prophylaxis it dropped significantly (P greater than 0.01) from the initial 3.16% AB.C. in March to 1.77% AB.C. in June, and in the group of coal cleaning plant workers (1.30% AB.C.) it was more or less equal to that of matching controls (1.21% AB.C.). Implicitly, coal dust alone can hardly be made responsible for the rise in % AB.C. It appears that the increase in frequency of chromosome aberrations observed in soft coal open-cast mining workers rather reflects an increased risk of genetic injury due to exposure to ever-present fumes and fire hazards that may give rise to the mutagenic chemicals belonging to a class of polycyclic aromatic hydrocarbons. The antimutagenic effect of ascorbic acid prophylaxis was characterized by a significant drop in % AB.C. and a speed-up of the lymphocyte cell cycle expressed as the proliferating rate index (PRI) whose value rose from the original 1.41 to 1.69, thus reaching the PRI value of matching controls (1.63).     

Distribution of sialic acid between sialoglycoproteins and other membrane components of different erythrocyte phenotypes.

The sialic acid content of erythrocytes of three different AB0 blood groups have been studied. The sialic acid contents of erythrocyte membranes containing 300 mg protein were determined and compared. Groups 0 (Rhesus negative), AB (both Rhesus negative and positive), and B (Rhesus negative) blood differed significantly (p less than 0.05) in total sialic acid content and in the distribution of sialic acid between sialoglycoproteins and other membrane components. Membrane materials containing 300 mg total protein showed sialic acid contents of 52.73 +/- 2.2 mumol sialic acid for group 0 (Rhesus negative) 34.77 +/- 1.16 mumol for group AB (Rh negative), 32.88 +/- 1.52 mumol for AB (Rh positive) and 21.23 +/- 0.84 mumol for B (Rh negative). In group 0 (Rh. neg.) membranes 39.4 +/- 1.4% of the total sialic acid was associated with the sialoglycoproteins. The percentage of sialic acids associated with sialoglycoproteins in other erythrocyte membranes were 77.7 +/- 1.3% for group B, and 55.6 +/- 1.0% and 56.4 +/- 1.8% for group AB (Rh. negative) and (Rh. positive) respectively. The changes appear to be independent of the Rhesus grouping but dependent on the AB0 grouping since membranes of the two Rhesus types of group AB had identical total sialic acid and sialoglycoproteins sialic acids. The sialic acid densities in sialoglycoproteins also differed from one erythrocyte type to another. Group 0 (Rh. negative) membrane sialoglycoproteins had sialic acid density of 140.5 +/- 3.1 nmol/mg compared to 71.7 +/- 1.2 nmol/mg for group B and 128.1 +/- 2.2 and 124.5 +/- 4.0 nmol/mg for group AB Rhesus negative and Rhesus positive respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chromosomal aberrations, SCE and urine mutagenicity in workers occupationally exposed to cytostatic drugs

The genetic risk run by workers occupationally exposed to chemicals, including the newly developed cytostatics, was assessed in large chemical laboratories. The exposed group comprising 38 people consisted of chemists, laboratory assistants and pilot plant workers. The average rate of aberrant cells in peripheral blood lymphocytes (AB.C). was 3.9% and the value of break/cell ratio was 0.046. The group of matched controls (N = 18) was found to have 1.5% of AB.C. and 0.020 breaks/cell. In the exposed group, there were, on average, 8.94 SCEs/cell, and in controls, 5.81. Urine mutagenicity tests on Salmonella typhimurium tester strain TA98 revealed a significant increase of activity in 23 (64%) out of 36 urine samples tested as compared to the control group (N = 19) with 4 (21%) positive urine samples. Strain TA100 showed only a weak response to urine mutagens.     

Chromosomal aberrations and sister-chromatid exchanges in swine

Frequencies of chromosomal aberrations and sister-chromatid exchanges (SCEs) in peripheral blood lymphocytes were investigated in 56 swine from 3 herds (I, breeding sows; II and III, fattening pigs). The mean frequencies of aberrant cells (AB.C.) were 3.58 +/- 1.59%, 2.10 +/- 1.52% and 6.20 +/- 3.21%, respectively. The mean numbers of SCEs per cell were 7.73 +/- 0.86, 6.51 +/- 0.89 and 7.06 +/- 1.47, respectively. A significant difference was found between the herds under study with regard to the number of aberrant cells but not the SCE frequency. In a parallel study, the presence of aflatoxin B1, polychlorinated biphenyls (PCB), dichlorodiphenyltrichloromethylmethane (DDT), lindane, mercury, lead and cadmium in the environment of fattening pigs was investigated. The total exposure to mutagens of pigs from herd III with a mean frequency of 6.2% AB.C., was markedly higher than that of herd II with the mean frequency of 2.1% AB.C.     

Possible genetic damage in the Czech nuclear power plant workers

The aim of our study was to identify occupational risk of irradiation exposure in the Czech nuclear power plant workers. We analyzed levels of chromosomal aberrations, a well-known biomarker of early biological effects and a predictor of cancer risk. We applied the conventional method of cytogenetic analysis and fluorescence in situ hybridization (FISH, whole chromosome painting for chromosomes 1 and 4, combined with a pancentromeric probe) to three groups: 123 subjects in the Temelin nuclear power plant (2 years in use), 114 subjects in the Dukovany nuclear power plant (20 years in use), and 53 matched controls from Ceske Budejovice. Nuclear power plant workers were divided into two groups: subjects with admittance into the monitored zone, and others. Following factors were also analyzed: GSTM1, GSTT1, GSTP1, XPD, XRCC1, hOGG1, p53, MTHFR, and MS gene polymorphisms, levels of vitamins A, C, E, and folate in plasma, and level of cotinine in urine. Long-term exposure to ionizing radiation in the monitored zone was 0.47+/-1.50 mSv (miliSievert) in the Temelin nuclear power plant and 5.74+/-9.57 mSv in the Dukovany nuclear power plant. Using the conventional cytogenetic analysis, we observed 1.90+/-0.95 and 1.82+/-1.19% AB.C. (percent of aberrant cells) in the Temelin nuclear power plant, and 2.39+/-1.01 and 2.33+/-1.04% AB.C. in the Dukovany nuclear power plant, for monitored zone workers and others, respectively. In the control group, we found 2.25+/-0.82% AB.C. Genomic frequency of translocations F(G)/100 measured by FISH was 1.89+/-1.40 and 2.01+/-1.68 in the Temelin nuclear power plant, and 2.48+/-1.93 and 2.14+/-1.62 in the Dukovany nuclear power plant for monitored zone workers and others, respectively. In the control group, F(G)/100 was 1.83+/-1.19. Following factors were identified as potential confounders by the conventional cytogenetic analysis: XPD-6, by the FISH: age, GSTP1 and p53Bst genotypes, long-term use of medication, alcohol consumption, and smoking. No association between the dose of irradiation and the level of chromosomal aberrations in any nuclear power plant was detected either by the conventional cytogenetic analysis or by FISH.     

Leptin response to short-term fasting in sympathectomized men: role of the SNS

We studied plasma leptin levels in six people with high-lesion spinal cord injury [SCI; body mass index (BMI) 25.9 +/- 1.5 kg/m(2), age 37 +/- 3.0 yr] and six able-bodied (AB) controls (BMI 29.1 +/- 1.9 kg/m(2), age 35 +/- 3.5 yr) before and after 12, 24, and 36 h of fasting. The plasma leptin levels significantly decreased during 36 h fasting by 48.8 +/- 4.5% (pre: 11.3 +/- 2.3, post: 6.2 +/- 1.5 ng/ml) and 38.6 +/- 7.9% (pre: 7.6 +/- 5.0, post: 4.2 +/- 1.0 ng/ml) in SCI and AB, respectively. Plasma leptin started to decrease at 24 h of fasting in the SCI group, whereas plasma leptin started to decrease at 12 h of fasting in the AB group. The current study demonstrated that plasma leptin decreased with fasting in both SCI and AB groups, with the leptin decrease being delayed in the SCI group. The delayed leptin response to fasting in the SCI group may be because of increased fat mass (%body fat, SCI: 33.8 +/- 3.0, AB: 24.1 +/- 2.9) and sympathetic nervous system dysfunction.     

Correlation of human lymphocyte SCE frequency with smoking history

The relationship between the level of occupational exposure to epichlorohydrin (ECHH) and the clastogenic effect was studied on a group of 33 workers. The effect of ECHH was assessed by differences in the frequency of chromosome aberrations in peripheral blood lymphocytes in ECHH-exposed and control groups. In the group exposed to the average ECHH concentration, 0.384 mg X m-3, during the last 6 months, the cytogenetic analysis revealed 2.00 +/- 0.23% AC (aberrant cells) (0.0203 B/C, breaks per cell) as compared with 1.68 +/- 0.23% AC (0.0172 B/C) in the matching controls. These results indicate that an average concentration lower than 0.40 mg X m-3 ECHH in the working atmosphere has no significant clastogenic effect on human peripheral lymphocytes.     

Adherence of peripheral blood neutrophils in X-ray operators.

The studies were conducted among 39 people (29 women and 10 men) operating X-ray equipment in radiology institutes and comparisons were made in the following schemes: A/ Group of persons operating the X-ray equipment (39 persons) and control group (18 persons). B/ A subgroup of persons that had been operating radiology equipment for > 5 years (29 workers was separated) and the results were compared with those for the subgroup of persons working < 5 years (10 workers). C/ Among the workers a subgroup of women (29) was separated and the results were compared with those for the subgroup of men (10 men). The following investigations were carried out: 1./ The absolute number of neutrophils in peripheral blood. 2./ Neutrophil adherence to fibres. 3./ Evaluation of CD 11b/CD 18 adhesion molecules expression on the surface of neutrophils. 4./ Evaluation of fibronectin concentration in serum. In the group A of the employees examined who operated X-ray units compared with the control group, the statistically significant decrease in the percentage of peripheral blood neutrophils showing adherence was observed and statistically significant decrease in median value and decrease of "Peak" value from the curve of the CD 18 adhesion molecule expression was detected and amounted to respectively: 91.0 +/- 4.26% adhering neutrophils in workers operating X-ray units and 94.8 +/- 2.98% in control group; values of median 120 +/- 4 in workers and 123 +/- 3 in control group in logarithmic scale; values of "Peak" 120 +/- 4 in workers and 123 +/- 3 in control group in logarithmic scale. Our own testing results show that the adherence properties of the peripheral blood neutrophils taken from X-ray operators are independent of the length of employment period and of the sex of the persons tested.     

Mutagenicity studies on paracetamol in human volunteers. I. Cytogenetic analysis of peripheral lymphocytes and lipid peroxidation in plasma

The clastogenic activity of paracetamol (PC) was assayed on a group of 11 healthy volunteers. PC was administered in the form of tablets 3 x 1000 mg in the course of 8 h. Blood samples were taken 0, 24, 72 and 168 h after the first application of the drug. Each blood sample was used for the cytogenetic analysis of peripheral lymphocytes, for the measuring of the level of lipid peroxidation (LPO) and ascorbemia in plasma. After PC administration the frequency of aberrant cells (AB.C.) was increased to 2.77% AB.C. after 24 h vs. 1.68% at 0 h, and breaks per cell (B/C) to 0.0295 vs. 0.0182, respectively. If PC was applied simultaneously with ascorbic acid (AA), also in a dose of 3 x 1000 mg, an increased frequency of AB.C. was observed only after 72 h, of B/C after both 24 h and 72 h. No increase in LPO as determined by the thiobarbituric acid assay was seen after PC administration (1.02-1.10 nmole malondialdehyde (MDA)/ml plasma). The LPO level was increased 72 h after the simultaneous application of PC and AA (1.26 nmole MDA/ml). No effect of AA in terms of a decreased PC clastogenicity was observed.     

Effect of ascorbic acid prophylaxis on the frequency of chromosome aberrations, urine mutagenicity and nucleolus test in workers occupationally exposed to cytostatic drugs

The genetic risk of workers occupationally exposed to a series of newly developed cytostatic drugs and the presumed antimutagenic potential of ascorbic acid (AA) were studied in a group of 38 chemical laboratory personnel examined for chromosome aberrations in lymphocytes, urine mutagenicity and nucleolar RNA activity before and after a 6-month prophylactic administration of AA at daily doses of 1 g for 5 days a week. Chromosome aberration tests revealed elevated aberrant cell (AB.C) rates both prior to and after AA supplementation (3.9% and 3.65% of AB.C., respectively). These values were significantly higher than those found in 18 non-exposed matching controls (1.05% of AB.C.). Tests for mutagenic activity in the urine of drug-exposed workers revealed 64% positive urine samples prior to vitaminization and 60% positive urine specimens after it; positive urine samples in the group of controls accounted for 21% of samples. In the nucleolus test, numbers of inactivated micronuclei in the exposed were initially higher than those of controls (33.4% versus 24.3%), but dropped to 20.5% after AA supplementation. These findings show that AA prophylaxis alone cannot substantially reduce the hazards associated with exposure to anti-cancer drugs.     

17beta-estradiol regulation of human growth hormone (hGH), insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3) axis in hypoestrogenic, hypergonadotropic women

OBJECTIVE: Ovarian hormonal function may be as important contributing factor to hGH-IGF-I-IGFBP-3 axis as age. AIM: To examine plasma hGH, IGF-1 and IGFBP-3 levels in women with premature ovarian failure compared to healthy normal controls and postmenopausal ones. PATIENTS: Group A-15 women with premature ovarian failure (POF) (mean: age 38.9+/-5.2 years, FSH 101.4+/-29.0 IU/l; 17beta-estradiol 22.5+/-14.6 ng/l). Group B consisted of 15 menopausal women (mean: age 54.7+/-2.7 years; FSH 81.9+/-32.1 IU/l; 17beta-estradiol 17.1+/- 8.0 ng/l). Group C - controls - 15 normally menstruating women (mean: age 37.1+/-9.0 years; FSH 6.2+/-1.0 IU/l; 17beta-estradiol 144.8+/-117.1 ng/l). METHODS: Body mass and BMI were measured. Basic fasting plasma hGH, IGF-I, IGFBP-3, insulin, testosterone and LH as well as prolactin (PRL), FSH and estradiol were assessed by RIA kits. Statistical analysis. Shapiro-Wilk test, Mann-Whitney u-test, Spearman rang correlation coefficient, stepwise multiple regression. RESULTS: Mean serum IGF-I level was the lowest (p<0.005) in group B (172.0+/-54.6 microg/l) and the highest in group C (273.6+/-109.0 microg/l). The mean plasma IGF-I level in group A was similar (NS) (208.3+/-66.5 microg/l) to that found in group B and lower (p<0.02) compared with that in group C. The lowest (p<0.005) serum IGFBP-3 level was found in group B (3.1+/-0.7 microg/l) compared to group C (4.4+/-0.3 microg/l). The mean plasma IGFBP-3 level (3.1+/-1.0 microg/l) in group A was lower than in group C (p<0.005) but identical as in group B. No statistically significant differences between groups were observed in mean hGH levels. Women in group A and C were younger (p<0.001) than those in group B. The lowest mean estradiol level was found in groups A and B. The highest was in group C (p<0.001). Mean plasma LH and FSH levels were higher (p<0.001) in groups A and B vs group C. In group C there were links between IGF-I and age (r=-0.60; p=0.014) The IGF-I/age relation disappeared in the groups A and B (rA=-0.26; rB=0.10; NS). The same regards IGFBP-3/ age link (rA=-0.44, NS; rB=0,31;NS). Estradiol level was related to hGH levels in group C (r=-0.54; p<0.05). In none of groups hGH/IGF-1 as well as IGFBP-3/hGH relations were found. Prolactin accounted for 69% of the variance in IGF-I level in the group B (p=0.003) and for 24% in group A (NS). Testosterone accounted for 88% (p=0.004) of the variance in IGF-I level in group B and IGFBP-3 was responsible for 86% (p=0.038) of the variance in IGF-I level in group C. Again IGFBP-3 was responsible for 47% (p=0.023) in group A and for 49% (p=0.04) in group B of the hGH variance. CONCLUSIONS: 17b-estradiol may be as important contributor to insulin-like growth factor-I (IGF-I) plasma level as age in hypoestrogenic, hypogonadotropic women.     

Effects of recombinant human extracellular-superoxide dismutase type C on myocardial reperfusion injury in isolated cold-arrested rat hearts.

The efficacy of recombinant human extracellular-superoxide dismutase type C (EC-SOD C) on myocardial reperfusion injury was explored in hypothermically arrested rat hearts, as was its site of action. Forty isolated working rat hearts were subjected to 30 min of global ischemia followed by 30 min of reperfusion. The hearts were arrested by the administration of 10 mL of cold perfusate at the onset of ischemia. At the same time, they were randomly assigned to one of five groups; A: cold perfusate only; B: cold perfusate + EC-SOD C 10.4 mg/L (30,000 U/L); C: cold perfusate+bovine CuZn-SOD 7.5 mg/L (30,000 U/L); D: cold perfusate + EC-SOD C 10.4 mg/L + heparin 50,000U/L; E: cold perfusate + heparin 50,000 U/L. Heparin was given to prevent binding of EC-SOD C to endothelial cell surfaces. Left ventricular function was studied before ischemia and at the end of reperfusion. Percent recovery of maximal left ventricular dP/dt after reperfusion was more pronounced in group B (109 +/- 24%; p less than .05) than in groups A (42 +/- 40%), C (47 +/- 36%), D (44 +/- 33%) and E (58 +/- 25%). Likewise, percent recovery of the double product (heart rate x systolic left ventricular pressure) was better in group B (104 +/- 18%; p less than .05) than in the other groups (A: 47 +/- 37%, C: 49 +/- 36%, D: 50 +/- 35%, E: 69 +/- 31%). Compared to the preischemic level, creatine kinase increased significantly in the coronary effluent after reperfusion in groups A, C, D, and E, but not in group B. The results suggest that EC-SOD C, which attaches to the endothelial cell surfaces, might be particularly effective as protection against myocardial reperfusion injury when given together with cardioplegic solution.     

Immunobiochemical profiles of workers differing in the degree of occupational exposure to vinyl chloride

The immunobiochemical studies were conducted in a group of 98 production workers engaged in polyvinyl chloride manufacture from ethylene (group A workers) and in a group of 59 vinyl chloride workers from a chemical plant employing classic production technology from acetylene (group B workers). Both groups of workers were matched by age (group A workers: 37.7 +/- 8.66 years; group B workers: 34.9 +/- 11.2 years) and average exposure length (group A workers: 8.6 +/- 3.0 years; group B workers: 10.7 +/- 8.4 years). All workers were examined for the serum concentrations of immunoglobulins IgG, IgA and IgM and acute reactants lysozyme (LYS), transferrin (TRF), ceruloplasmin (CPL), alpha-l-antitrypsin (AlAT), alpha-2-macroglobulin (A2M) and orosomucoid (ORO). The statistical analysis included calculations of means, standard deviations and 95% confidence intervals. Differences in means were evaluated by t-test, differences in the distribution pattern of values by F-test. Abnormality of values was assessed by comparisons to normal values valid in Czechoslovakia. Group A worked in conditions meeting the MAC 10 mg VC.m-3 comparing with group B workers had elevated levels of IgG (P less than 0.005), IgA and IgM (P less than 0.001 both). Group B workers differed from group A workers by exhibiting significantly elevated levels of AlAT, and CPL. (P less than 0.001). The differences in the frequency of abnormal values between group A and group B worked in substantially less favourable hygienic conditions were significant for immunoglobulins elevated in group A and for ORO (P less than 0.01) and CPL (P less than 0.001) elevated in group B. The possible relationship of these immunobiochemical findings with the degree of vinyl chloride exposure are critically analyzed.     

The effects of exposure to different clastogens on the pattern of chromosomal aberrations detected by FISH whole chromosome painting in occupationally exposed individuals

The pattern of chromosomal aberrations (CA) was studied by fluorescence in situ hybridization (FISH) technique (whole chromosomes #1 and #4 painting) in workers occupationally exposed to any of the four following conditions: acrylonitrile (ACN), ethyl benzene (EB), carcinogenic polycyclic aromatic hydrocarbons (c-PAHs), and irradiation in nuclear power plants (NPP), respectively. Decrease in the relative frequency of translocations was observed in EB group, and an increase in reciprocal translocations in ACN and NPP-exposed groups. An increase in a relative number of insertions was registered under all four conditions (significant at ACN, EB, c-PAHs, quasisignificant at NPP-exposed groups). Significant differences in the percentage of lymphocytes with aberrations on chromosome #1 (58.8+/-32.7%, versus 73.8+/-33.6% in the controls, P < 0.05), and chromosome #4 (47.0+/-34.1%, versus 29.4+/-32.2%, P < 0.01) were found in workers exposed to ACN. Similarly, a decrease in the proportion of cells with aberration on chromosome #1 (61.0+/-24.0%, versus 73.8+/-33.6%, P < 0.05) and an increase on chromosome #4 (45.6+/-24.6%, versus 29.4+/-32.2%, P < 0.05) were observed in workers exposed to EB. Frequency of aberrant cells (%AB.C.) as well as genomic frequency of translocations (F(G)/100) increased with age (P < 0.001). Aging also increased the percentage of translocations and reciprocal translocations (P < 0.05), but decreased the relative number of acentric fragments (P < 0.01). Smoking led to significantly increased F(G)/100 (P < 0.05), but did not affect the pattern of chromosomal aberrations. Our results seem to indicate that different carcinogens may induce a different pattern of chromosomal aberrations.     

Effect of fish oil diet on hepatic lipid metabolism in nonhuman primates: lowering of secretion of hepatic triglyceride but not apoB

African green monkeys were fed diets containing either 11% (by weight) fish oil or lard for 2.5 yr. To test the hypothesis that fish oil decreases hepatic secretion of triglyceride (TG) and apoB, livers from these animals were perfused with a fatty acid mixture [85% (w/w) oleate containing [14C]oleate and 15% n-3 containing [3H]eicosapentaenoic acid (EPA)] at a rate of 0.1 mumol fatty acid/min per g liver. Liver perfusate was sampled every 30 min during 4 h of recirculating perfusion. The concentration of triglyceride was similar for livers of animals of both groups and there was no difference between groups in the extent of incorporation of [3H]EPA or [14C]oleate into hepatic TG. While the secretion rate for the mass of TG was less in the fish oil-fed group (8.3 +/- 2.5 vs 18.3 +/- 4.4 mg/h per 100 g liver, P less than 0.05), the apoB secretion rate was similar (0.92 +/- 0.15 vs 1.01 +/- 0.13 mg/h per 100 g liver). Significantly less [3H]EPA was incorporated into secreted TG in the fish oil group (0.4 +/- 0.1 vs 1.0 +/- 0.1% infused dose/h; P less than 0.01). The rate of secretion of [14C]TG was similar for both groups (1.3 +/- 0.3 vs 1.4 +/- 0.1% infused dose/h for fish oil and lard groups, respectively). No significant diet-related differences in [3H]TG or [14C]TG fatty acid specific activity were observed for perfusate TG or hepatic TG. After perfusion, livers from fish oil-fed monkeys contained significantly more [3H]EPA in hepatic phospholipid than livers from lard-fed monkeys (19.5 +/- 1.8 vs 11.4 +/- 1.7% infused dose; P less than 0.01) although hepatic phospholipid mass concentrations were similar. The liver phospholipids of the fish oil group were enriched in n-3 fatty acid mass and were relatively depleted of oleate and linoleate. We conclude that although apoB secretion was unaffected, dietary fish oil significantly decreased hepatic TG secretion through relatively poor utilization of EPA for the synthesis of TG destined for secretion in VLDL; at the same time, increased incorporation of [3H]EPA into hepatic phospholipid accompanied the decreased incorporation into secreted TG and these events may be coupled.(ABSTRACT TRUNCATED AT 400 WORDS)     

Regulation of idiotope expression. III. H-2 influences the magnitude and the idiotypy of a T-independent antibody response in mice of certain genetic backgrounds

Antibody response to the phosphocholine (PC) epitope on Streptococcus pneumoniae R36a (Pn), a T-independent Ag type 2, was studied in H-2 congenic mouse strains. The PC-specific antibody plaque-forming cells (PFC) were enumerated in the spleen at various intervals after the primary Pn injection, and the proportion of PFC that produced antibody expressing the AB1-2 idiotope (Id) was determined by using the corresponding monoclonal anti-Id. AB1-2 is a cross-reactive Id, detectable on germline-encoded PC antibody of the T15 family, and on most, but not all, somatic variants of that antibody. The specific PFC responses in BALB/c (H-2d) and BALB.B (H-2b) strains were of comparable magnitude and most, if not all, PFC were ABl-1 Id-positive (AB1-2+). This was not the case in the responses of the B10D2 (H-2d) vs C57BL/10 (H-2b) strains and the D1.C (H-2d) vs D1.LP (H-2b) strains (on DBA/1 background). In each of these pairs, the H-2d mice were high responders, and the response was dominated by AB1-2 Id (greater than or equal to 80% AB1-2+ PFC at the peak, on day 5). The H-2b mice were low responders, and only a minor proportion of PFC (less than or equal to 30%) were AB1-2+; an increase of AB1-2+ was seen later in the response (d.10). The results of PFC assays were confirmed by measuring the PC-binding antibody and AB1-2 Id in the sera of D1.C and D1.LP mice immunized repeatedly with Pn. Moreover, D1.LP mice that had very low levels of AB1-2 Id had higher serum levels of antibody expressing two other T15 Id, B36-82, and B24-44. The B36-82 and B24-44 Id have been previously found on somatic variants of PC antibody expressed independently of the Ab1-2 Id. The concentrations of these two Id in D1.LP mice after repeated immunization approached those in D1.C. These results indicate that 1) the H-2 allelism may have a significant effect on TID antibody response in mice of a certain genetic background, but not in the BALB/c; and 2) the idiotypic repertoire of the response may be influenced by H-2 at the level of clonal variants of PC-reactive cells.     

Optical measurement of osmotic water transport in cultured cells. Role of glucose transporters

Methodology was developed to measure osmotic water permeability in monolayer cultured cells and applied to examine the proposed role of glucose transporters in the water pathway (1989. Proc. Natl. Acad. Sci. USA. 86:8397-8401). J774 macrophages were grown on glass coverslips and mounted in a channel-type perfusion chamber for rapid fluid exchange without cell detachment. Relative cell volume was measured by 45 degrees light scattering using an inverted microscope; measurement accuracy was validated by confocal imaging microscopy. The time required for greater than 90% fluid exchange was less than 1 s. In response to a decrease in perfusate osmolality from 300 to 210 mosM, cells swelled without lag at an initial rate of 4.5%/s, corresponding to a water permeability coefficient of (6.3 +/- 0.4) x 10(-3) cm/s (SE, n = 20, 23 degrees C), assuming a cell surface-to-volume ratio of 4,400 cm-1. The initial rate of cell swelling was proportional to osmotic gradient size, independent of perfusate viscosity, and increased by amphotericin B (25 micrograms/ml), and had an activation energy of 10.0 +/- 1 kcal/mol (12-39 degrees C). The compounds phloretin (20 microM) and cytochalasin B (2.5 micrograms/ml) inhibited glucose transport by greater than 85% but did not influence Pf in paired experiments in which Pf was measured before and after inhibitor addition. The mercurials HgCl2 (0.1 mM) and p-chloromercuribenzoate (1 mM) did not inhibit Pf. A stopped-flow light scattering technique was used to measure Pf independently in J774 macrophages grown in suspension culture. Pf in suspended cells was (4.4 +/- 0.3) x 10(-3) cm/s (assuming a surface-to-volume ratio of 8,800 cm-1), increased more than threefold by amphotericin B, and not inhibited by phloretin and cytochalasin B under conditions of strong inhibition of glucose transport. The glucose reflection coefficient was 0.98 +/- 0.03 as measured by induced osmosis, assuming a unity reflection coefficient for sucrose. These results establish a quantitative method for measurement of osmotic water transport in adherent cultured cells and provide evidence that glucose transporters are not involved in the water transporting pathway.     

Fish oil induced increase in walking distance, but not ankle brachial pressure index, in peripheral arterial disease is dependent on both body mass index and inflammatory genotype

Peripheral arterial disease (PAD) is an atherosclerotic disease. Evidence suggests that atherosclerosis is an inflammatory condition and long chain n-3 fatty acids, found in oily fish and fish oils, have been shown to reduce inflammation. Genetic and lifestyle factors such as body mass index (BMI) also influence inflammation. In this study we have examined the effect of fish oil in patients with claudication secondary to PAD. Fish oil supplementation, providing 1g EPA and 0.7 g DHA per day for 12 weeks, increased walking distance on a treadmill set at 3.2 km/h with a 7% incline. Walking distance to first pain increased from 76.2+/-8.5 m before fish oil to 140.6+/-25.5 m after fish oil (mean+/-SEM, p=0.004) and total distance walked increased from 160.0+/-21.5 m before fish oil to 242.1+/-34.5 m after fish oil (p=0.002). Fish oil supplementation also improved ankle brachial pressure index (ABPI) from 0.599+/-0.017 before fish oil to 0.776+/-0.030 after fish oil (p<0.001). The increase in walking distance was dependent on both BMI and genotype for single nucleotide polymorphisms in the genes encoding the pro-inflammatory cytokines tumour necrosis factor-alpha and interleukin (IL)-1beta and the anti-inflammatory cytokine IL-10 (detected using amplification refractory mutation system polymerase chain reaction). Neither BMI nor any of the genotypes examined affected the ability of fish oil to increase ABPI. The mechanisms by which fish oil affects walking distance and ABPI do not appear to be the same.     

Omega-3 fatty acid supplementation accelerates chylomicron triglyceride clearance

Omega-3 fatty acids (FAs) reduce postprandial triacylglycerol (TG) concentrations. This study was undertaken to determine whether this effect was due to reduced production or increased clearance of chylomicrons. Healthy subjects (n = 33) began with a 4-week, olive oil placebo (4 g/d) run-in period. After a 4-week wash-out period, subjects were randomized to supplementation with 4 g/d of ethyl esters of either safflower oil (SAF), eicosapentaenoic acid (EPA), or docosahexaenoic acid (DHA) for 4 weeks. Results for EPA and DHA were similar, and therefore the data were combined into one omega-3 FA group. Omega-3 FA supplementation reduced the postprandial TG and apolipoprotein B (apo B)-48 and apoB-100 concentrations by 16% (P = 0.08), 28% (P < 0.001), and 24% (P < 0.01), respectively. Chylomicron TG half-lives in the fed state were reduced after omega-3 FA treatment (6.0 +/- 0.5 vs. 5.1 +/- 0.4 min; P < 0.05), but not after SAF (6.9 +/- 0.7 vs. 7.1 +/- 0.7 min). Omega-3 FA supplementation decreased chylomicron particle sizes (mean diameter; 293 +/- 44 vs. 175 +/- 25 nm; P < 0.01) and increased preheparin lipoprotein lipase (LPL; 0.6 +/- 0.1 vs. 0.9 +/- 0.1 micromol/h/ml; P < 0.05) activity during the fed state, but had no effect on postheparin LPL or hepatic lipase activities. The results suggest that omega-3 FA supplementation accelerates chylomicron TG clearance by increasing LPL activity, and that EPA and DHA are equally effective.