Survival of density subpopulations of rabbit platelets: use of 51Cr-or 111In-labeled platelets to measure survival of least dense and most dense platelets concurrently

The origin of the density heterogeneity of platelets was studied by measuring the survival of density subpopulations of rabbit platelets separated by discontinuous Stractan density gradient centrifugation. When a total population of 51Cr-labeled platelets was injected into recipient rabbits, the relative specific radioactivity of the most dense platelets decreased rapidly. In contrast, that of the least dense platelets had not changed 24 hr after injection, and then decreased slowly. To distinguish between the possibilities that most dense platelets are cleared from the circulation more quickly than least dense platelets or that platelets decrease in density as they age in the circulation, the concurrent survival of least dense and most dense platelets, labeled with either 51Cr or 111In-labeled total platelet populations, determined concurrently in the same rabbits, were identical, calculated from 1 hr values as 100%. However, the 1-hr recovery of 111In-labeled platelets was slightly but significantly less than that of 51Cr-labeled platelets. Therefore, we studied the survival of 51Cr-labeled least dense and 111In-labeled most dense platelets as well as that of 111In-labeled least dense and 51Cr-labeled most dense platelets. Mean 1-hr recovery of least dense platelets, labeled with either isotope (78% +/- 7%, SD) was similar to that of most dense platelets, labeled with either isotope (77% +/- 8%; SD). Mean survival of least dense platelets was 47.3 +/- 18.7 hr (SD), which was significantly less than that of most dense platelets (76.1 +/- 21.6 hr; SD) (p less than 0.0025). These results indicate that platelets decrease in buoyant density as they age in the circulation and that most dense platelets are enriched in young platelets, and least dense in old. Thus, the events that affect platelets as they age in the circulation contribute to platelet density heterogeneity, although they may not be the sole cause of it.     

Inhibition of cyclic 3',5'-nucleotide phosphodiesterase-a possible mechanism of action of bifunctional alkylating agents.

The effect of anti-tumour alkylating agents on the adenosine 3′,5′-monophosphate (cyclic AMP) levels of sensitive and resistant Walker carcinoma cells in tissue culture has been investigated. Chlorambucil caused a two-fold elevation of cyclic AMP, in the sensitive line only, 8 hr after treatment with a dose of 5 μg/ml. A comparable increase is produced by the cyclic nucleotide phosphodiesterase inhibitor aminophylline at a dose which produces the same degree of inhibition of cell growth. The therapeutically-inactive monofunctional N-ethyl analogue of chlorambucil had no effect on the cyclic AMP level of the sensitive cells at a dose of 250 μg/ml after 8 hr, while the highly selective monofunctional alkylating agent, CB 1954, at 1 μg/ml caused an elevation of cyclic AMP in the sensitive line 24 hr after treatment. This is not a non-specific effect caused by an inhibition of cell growth for the antimetabolite methotrexate had no effect on the intracellular cyclic AMP of sensitive Walker cells at doses which produced complete inhibition of cell growth. The effect of the alkylating agents on cyclic AMP levels is probably due to a specific inhibition of the cyclic nucleotide phosphodiesterase with a low K_m value, since only this form of the enzyme is inhibited, and only in the sensitive cells, when an effect on cell growth and cyclic AMP content is observed. In the case of CB 1954, however, there was no inhibition of either form of the phosphodiesterase at a time when cyclic AMP levels were elevated. A linear relationship exists between the reciprocals of the intracellular cyclic AMP and the percentage growth inhibition produced by a given dose of chlorambucil.     

Patient information booklets for Asian patients with ulcerative colitis

Our aim was to address the information requirements for ulcerative colitis patients from Asian ethnic minorities in Leicester city. We sought to determine if the information leaflets provided in English could be successfully employed when translated into the common South Asian languages. A postal survey determined the initial demand for information leaflets, offering the leaflet in English, Hindi, Gujarati or Punjabi. Follow up questionnaires were again by post and subsequently by telephone contact.All patients found the leaflets useful, but felt that doctors should do more to help with language problems. On reading the leaflets, sixty-six percent of patients experienced reduced levels of anxiety or no change, whereas thirty-three percent found increased levels of anxiety. Nearly two-thirds of patients felt there was insufficient or satisfactory information in the leaflets. The final response rate for returning the questionnaire was 53%. The leaflets were generally well received, but there may be a role for increased detail, which may in turn reduce anxiety levels. The low response rates highlight the difficulty of communication with this group, suggesting that we need to make more resources available to these patients.     

Similar fecal immunochemical test results in screening and referral colorectal cancer

AIM: To improve the interpretation of fecal immunochemical test (FIT) results in colorectal cancer (CRC) cases from screening and referral cohorts. METHODS: In this comparative observational study, two prospective cohorts of CRC cases were compared. The first cohort was obtained from 10 322 average risk subjects invited for CRC screening with FIT, of which, only subjects with a positive FIT were referred for colonoscopy. The second cohort was obtained from 3637 subjects scheduled for elective colonoscopy with a positive FIT result. The same FIT and positivity threshold (OC sensor; ≥ 50 ng/mL) was used in both cohorts. Colonoscopy was performed in all referral subjects and in FIT positive screening subjects. All CRC cases were selected from both cohorts. Outcome measurements were mean FIT results and FIT scores per tissue tumor stage (T stage). RESULTS: One hundred and eighteen patients with CRC were included in the present study: 28 cases obtained from the screening cohort (64% male; mean age 65 years, SD 6.5) and 90 cases obtained from the referral cohort (58% male; mean age 69 years, SD 9.8). The mean FIT results found were higher in the referral cohort (829 ± 302 ng/mLvs 613 ± 368 ng/mL,P = 0.02). Tissue tumor stage (T stage) distribution was dif-ferent between both populations [screening population: 13 (46%) T1, eight (29%) T2, six (21%) T3, one (4%) T4 carcinoma; referral population: 12 (13%) T1, 22 (24%) T2, 52 (58%) T3, four (4%) T4 carcinoma], and higher T stage was significantly associated with higher FIT results (P 0.001). Per tumor stage, no significant difference in mean FIT results was observed (screening vs referral: T1 498 ± 382 ng/mL vs 725 ± 374 ng/mL, P = 0.22; T2 787 ± 303 ng/mL vs 794 ± 341 ng/mL, P = 0.79; T3 563 ± 368 ng/mLvs 870 ± 258 ng/mL,P = 0.13; T4 not available). After correction for T stage in logistic regression analysis, no significant differences in mean FIT results were observed between both types of cohorts (P = 0.10). CONCLUSION: Differences in T stage distribution largely explain differences in FIT results between screening and referral cohorts. Therefore, FIT results should be reported according to T stage.     

Plasma bile acids are not associated with energy metabolism in humans

Bile acids (BA) have recently been shown to increase energy expenditure in mice, but this concept has not been tested in humans. Therefore, we investigated the relationship between plasma BA levels and energy expenditure in humans. Type 2 diabetic (T2DM) patients (n = 12) and gender, age and BMI-matched healthy controls (n = 12) were studied before and after 8 weeks of treatment with a BA sequestrant. In addition, patients with liver cirrhosis (n = 46) were investigated, since these display elevated plasma BA together with increased energy expenditure. This group was compared to gender-, age- and BMI-matched healthy controls (n = 20). Fasting plasma levels of total BA and individual BA species as well as resting energy expenditure were determined. In response to treatment with the BA sequestrant, plasma deoxycholic acid (DCA) levels decreased in controls (-60%, p < 0.05) and T2DM (-32%, p < 0.05), while chenodeoxycholic acid (CDCA) decreased in controls only (-33%, p < 0.05). Energy expenditure did not differ between T2DM and controls at baseline and, in contrast to plasma BA levels, was unaffected by treatment with the BA sequestrant. Total BA as well as individual BA species did not correlate with energy expenditure at any time throughout the study. Patients with cirrhosis displayed on average an increase in energy expenditure of 18% compared to values predicted by the Harris-Benedict equation, and plasma levels of total BA (up to 12-fold) and individual BA (up to 20-fold) were increased over a wide range. However, neither total nor individual plasma BA levels correlated with energy expenditure. In addition, energy expenditure was identical in patients with a cholestatic versus a non-cholestatic origin of liver disease while plasma total BA levels differed four-fold between the groups. In conclusion, in the various (patho)physiological conditions studied, plasma BA levels were not associated with changes in energy expenditure. Therefore, our data do not support an important role of circulating BA in the control of human energy metabolism.