(i) Hepatocytes isolated from adult rats were cultured for 2 to 3 weeks on collagen in a modified, serum-free Waymouth medium containing fatty acids and varying concentrations of glucocorticoid, insulin and glucagon. (ii) In the presence of all three hormones, it was possible to maintain the content of DNA, the activity of glucokinase, pyruvate kinase, hexokinase and lactate dehydrogenase at initial levels for 2 to 3 weeks. The activity of glucokinase and pyruvate kinase was affected by the concentration of insulin. (iii) The activity of alcohol dehydrogenase was stable for 3 days and declined to about 25% of the initial level after 2 weeks of culture, irrespective of the presence of hormones. (iv) Maintenance of albumin secretion was dependent on the presence of glucocorticoid, and glucocorticoid and insulin showed an additive or, at some time points, a synergistic effect on its secretion. (v) The content of cytochrome P-450 could be kept at 65% of the initial level, provided that a relatively high concentration of dexamethasone was present (10(-6) M). (vi) In the absence of hormones, urea synthesis was 70% of initial levels throughout the experimental period. With insulin and glucocorticoid present, a high concentration of glucagon (10(-8) M) was required to maintain the synthesis of urea at this level. (vii) It is concluded that hepatocyte cultures as described in the present study may be a useful, well-defined system for long-term metabolic, pharmacologic and toxicologic studies.

Alcohol Dehydrogenase↗

PubMedDate not supplied

Distribution of the CCR5 gene 32 base pair deletion and SDF1-3'A variant in healthy individuals from different populations.

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Base Pairing↗

PubMedDate not supplied

Effect of ethanol on fatty acid metabolism in cultured hepatocytes: dependency on incubation time and fatty acid concentration.

In a previous report it was shown that ethanol increases the rate of accumulation of triacylglycerol by 90% in hepatocytes in primary culture. This represents the first known suitable model for in vitro studies of the ethanol-induced fatty liver. The biochemical alterations causing this accumulation of triacylglycerol remain to be elucidated, however. In the present report it is shown that (1) the effect of ethanol exhibits a time lag of 6-9 hours (2) the increment in the content of triacylglycerol caused by ethanol is increased by increased concentrations of fatty acids (3) the fatty acid uptake is not affected by ethanol (4) fatty acid synthesis is inhibited 20% by ethanol (5) the contents of diacylglycerol and phospholipids are not affected by ethanol (6) addition of ethanol increases the cytosolic and mitochondrial redox levels. It is concluded that ethanol is likely to exert its effect on the accumulation of triacylglycerol by redistributing fatty acids between oxidation and triacylglycerol synthesis and/or between storage and secretion of triacylglycerol.

Animals↗

PubMedDate not supplied

On the inheritance of the Era red cell antigen.

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Blood Group Antigens↗

PubMedDate not supplied

Association between HLA-DR1 and -DR3 antigens and unexplained repeated miscarriage.

Few, mostly small, studies have investigated the distribution of HLA class II antigens among women with unexplained recurrent miscarriage. Although some studies have reported statistically significant associations between this syndrome and certain HLA-DR antigens--especially the -DR1 and -DR3 antigens--other studies have been unable to demonstrate such associations. For the present meta-analysis, 18 cross-sectional or case-control studies (published or unpublished) reporting on frequencies of HLA-DR1 and -DR3 antigens among Caucasian women with unexplained repeated miscarriage were identified by searching literature databases (MEDLINE and EMBASE), reading the references of identified studies, and by contacting researchers within the field. The studies comprised a total of 1508 patients. The methodological quality of most of the studies was low, especially because of small numbers of patients and because patients with only two miscarriages were included in many studies; this is defined as repeated miscarriage. The odds ratios of repeated miscarriage for the HLA-DR1 and -DR3 antigens were calculated for the individual studies and subsequently the pooled odds ratios for the studies were calculated. The combined odds ratio for HLA-DR1 was 1.29 [95% confidence interval (CI) = 1.05-1.58] (17 studies) which is statistically significant (P <0.05). The combined odds ratio for HLA-DR3 was 1.00 (95% CI 0.80-1.24) (18 studies), which is not significant. The results of the meta-analysis suggest that the HLA-DR antigen DR1 is associated with an increased susceptibility to unexplained repeated miscarriage.

Abortion, Habitual↗

PubMedDate not supplied

n-3 fatty acids and leukocyte chemotaxis. Effects in hyperlipidemia and dose-response studies in healthy men.

Dietary supplementation with n-3 polyunsaturated fatty acids (n-3 PUFAs) has been shown to inhibit neutrophil and monocyte chemotaxis in healthy subjects and, with respect to neutrophils, also in various patient groups. We studied the effect of dietary supplementation with n-3 PUFAs on monocyte and neutrophil chemotaxis in patients with hyperlipidemia. Chemotaxis was investigated with the under-agarose assay, using autologous serum and N-formyl-methionyl-leucyl-phenylalanine as chemoattractants. The patients were examined before and after 6 weeks of supplementation with 6 g n-3 PUFAs daily. Monocyte chemotaxis was reduced after n-3 PUFA supplementation in type IIa patients but was unaffected in patients with type IV hyperlipidemia. Furthermore, monocyte chemotaxis was increased in untreated type IIa patients compared with normocholesterolemic controls. We also studied the dose-response effects of n-3 PUFAs on monocyte and neutrophil chemotaxis in healthy men given 1.3, 4, and 9 g n-3 PUFAs daily for 6-week periods. Monocyte and neutrophil chemotaxis was reduced after n-3 PUFA supplements in a dose-dependent fashion, with the majority of the effect observed after the low dose. These results lend support to the notion of an antiatherosclerotic effect of n-3 PUFAs and may provide an explanation for the hitherto-unexplained effect of low doses of n-3 PUFAs in coronary heart disease.

Chemotaxis, Leukocyte↗

PubMedDate not supplied

Direct cell mediated lympholysis in man. Cellular cytotoxic capacity of 100 normal, healthy blood donors.

The cellualr lymphocytotoxic capacity of 100 normal, healthy blood donors were investigated by the Direct Cell Mediated Lympholysis (DCML) test. The scorings were found to be distributed according to the normal, exhibiting a mean value of 0.1 Cr-51 release %, and an upper 1% significance level of 10.1 release 1%. It is concluded that lymphocytes from the peripheral blood of blood donors may well be used as negative biological controls in DCML testing of immunologically primed patients (e.g. after allotransplantation).

Blood Donors↗

PubMedDate not supplied

Direct cell mediated lympholysis by peripheral blood lymphocytes from renal allograft recipients.

Investigations of 22 kidney allograft recipients by the Direct CML-test revealed coincidence between acute allograft rejection and a positive test. 15 patients exhibited acute rejection, 3 chronic, and 4 no rejection episodes. 4 patients with acute rejection were not detected as positive in Direct CML, while 3 patients were positive without acute rejection, one having a chronic rejection. The exact role of HL-A antigens for the specificity of a positive Direct CML could not be evaluated from this material. It is concluded that the Direct CML-test may be helpful for the diagnosis of acute rejection, but that it has only limited power as the only test of recipients' response to foreign tissue.

Acute Disease↗

PubMedDate not supplied

Influence of radiographic contrast media on phagocytosis.

To evaluate the influence of radiographic contrast media (CM) on human polymorphonuclear leucocytes (PML), the ability of these cells to ingest latex particles after in vitro exposure to five different radiographic contrast media was investigated. All CM inhibited the phagocytic properties of PML. The inhibition was dose dependent. The inhibitory effect was partly due to hyperosmolality but CM specific inhibition was also evident.

Contrast Media↗

PubMedDate not supplied

Metabolism of thapsigargin in rat hepatocytes.

The cytotoxicity, uptake, and metabolism of thapsigargin, an inhibitor of the Ca(2+)-ATPases of the sarco- and endoplasmatic reticulum (the SERCA-family), were investigated in suspensions of rat hepatocytes using [3H]thapsigargin labeled at C-8. No effect was observed on the lactate dehydrogenase leakage from the cells or in glucose formation when hepatocytes were incubated with 0.5-25 microM thapsigargin. At 25 microM [3H]thapsigargin the initial rate of uptake into the cells was 471 nmol/10(8) cells/min. Thapsigargin metabolism followed first-order kinetics, with an initial rate of metabolism at 25 microM of 65 nmol/10(8) cells/min. The much faster uptake than metabolism suggests that thapsigargin probably is bound to cellular proteins or trapped in cellular membranes. The metabolites were characterized by normal and reversed phase TLC and by positive and negative FAB/ms. Two labeled products were identified when cells were incubated with 25 microM [3H]thapsigargin. The first product formed was desoctanoyl-thapsigargin, followed by formation of desacyl-thapsigargin. Thapsigargin metabolism was strongly inhibited by diethyl p-nitrophenyl phosphate, indicating that the deacylation of the compound is catalyzed by microsomal carboxylesterases. Upon prolonged incubation, a volatile, tritiated compound appeared, possibly water, due to oxidation of the alcohol group at C-8 of thapsigargin.

Animals↗