In vitro and in vivo activity of murine lymphokine-activated killer cells after cryopreservation

The in vitro and in vivo effects of cryopreservation on the cytotoxic activity of murine lymphokine-activated killer (LAK) cells were studied. LAK cells were generated by incubation of spleen lymphocytes of BALB/c mice for 3 days with recombinant interleukin-2 (rIL-2) and subsequent cryopreservation. Cytotoxicity was determined in a 51Cr release assay. After thawing, cytotoxic activity was reduced (40.4% 51Cr release at an effector:target cell ratio of 40:1 as compared to 68.5% 51Cr release before freezing) and could be restored to precryopreserved levels by reincubation with rIL-2 for 2 days after thawing (78.8% 51Cr release). These cells were then tested in BALB/c mice injected with RAW 112 cells, a pre-B-cell lymphoma line. The results demonstrate that the survival rate of mice injected with cryopreserved and restimulated LAK cells (50% survival greater than 180 days after injection) did not differ significantly from that of mice injected with fresh unfrozen LAK cells (60% survival greater than 120 days, 50% survival greater than 180 days). Cryopreserved LAK cells have potential use in adoptive immunotherapy.     

Contribution of individual targets to the antitumor efficacy of the multitargeted receptor tyrosine kinase inhibitor SU11248

Recent achievements in the development of multitargeted molecular inhibitors necessitate a better understanding of the contribution of activity against individual targets to their efficacy. SU11248, a small-molecule inhibitor targeting class III/V receptor tyrosine kinases, including the platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) receptors, KIT and FLT3, exhibits direct effects on cancer cells as well as antiangiogenic activity. Here, we investigated the contributions of inhibiting individual SU11248 target receptors to its overall antitumor efficacy in tumor models representing diverse signaling paradigms. Consistent with previous results, SU11248 was highly efficacious (frequently cytoreductive) in all models tested. To elucidate the specific contributions of inhibition of PDGF and VEGF receptors to the in vivo efficacy of SU11248, we employed two selective inhibitors, SU10944 (VEGF receptor inhibitor) and Gleevec (PDGF receptor inhibitor). SU10944 alone induced a tumor growth delay in all models evaluated, consistent with a primarily antiangiogenic mode of action. In contrast, Gleevec resulted in modest growth inhibition in tumor models in which the cancer cells expressed its targets (PDGFRbeta and KIT), but was not efficacious against tumors not driven by these target receptor tyrosine kinases. Strikingly, in all but one tumor model evaluated, the antitumor efficacy of SU10944 combined with Gleevec was similar to that of single-agent SU11248, and was greatly superior to that of each compound alone, indicating that the antitumor potency of SU11248 in these models stems from combined inhibition of both PDGF and VEGF receptors. The one exception was a model driven by an activated mutant of FLT3, in which the activity of SU11248, which targets FLT3, was greater than that of SU10944 plus Gleevec. Moreover, SU10944 combined with Gleevec inhibited tumor neoangiogenesis to an extent comparable to that of SU11248. Thus, the potent efficacy of SU11248 in models representing diverse signaling paradigms results from simultaneous inhibition of individual target receptors expressed both in cancer cells and in the tumor neovasculature, supporting the hypothesis that multitargeted inhibitors have the cumulative antitumor efficacy of combined single-target inhibitors.     

Family history of myocardial infarction is an independent risk factor for venous thromboembolism: the Tromsø study

Summary. Background: Recent studies indicate that arterial cardiovascular diseases and venous thromboembolism (VTE) share common risk factors. A family history of myocardial infarction (MI) is a strong and independent risk factor for future MI. Objectives: The purpose of the present study was to determine the impact of cardiovascular risk factors, including family history of MI, on the incidence of VTE in a prospective, population‐based study. Patients and methods: Traditional cardiovascular risk factors and family history of MI were registered in 21 330 subjects, aged 25–96 years, enrolled in the Tromsø study in 1994–95. First‐lifetime VTE events during follow‐up were registered up to 1 September 2007. Results: There were 327 VTE events (1.40 per 1000 person‐years), 138 (42%) unprovoked, during a mean of 10.9 years of follow‐up. In age‐ and gender‐adjusted analysis, age [hazard ratio (HR) per decade, 1.97; 95% confidence interval (CI), 1.82–2.12], gender (men vs. women; HR, 1.25; 95% CI, 1.01–1.55), body mass index (BMI; HR per 3 kg m^?2, 1.21; 95% CI, 1.13–1.31), and family history of MI (HR, 1.31; 95% CI, 1.04–1.65) were significantly associated with VTE. Family history of MI remained a significant risk factor for total VTE (HR, 1.27; 95% CI, 1.01–1.60) and unprovoked VTE (HR, 1.46; 95% CI, 1.03–2.07) in multivariable analysis. Blood pressure, total cholesterol, HDL‐cholesterol, triglycerides, and smoking were not independently associated with total VTE. Conclusions: Family history of MI is a risk factor for both MI and VTE, and provides further evidence of a link between venous and arterial thrombosis.     

Lack of effect of somatostatin on epinephrine-stimulated glucose production in the dog

The effects of somatostatin on epinephrine-stimulated hepatic glucose production were assessed in the conscious overnight fasted dog. Glucose production was measured using a primed constant infusion of 3-3H-glucose. Two experiments were performed on each of four animals, each experiment consisting of a tracer equilibration period (80 minutes), a control period (40 minutes), and a test period (180 minutes). In the first experiment, epinephrine was infused at 0.08 microgram/kg/min during the test period so that the plasma concentration rose from 138 +/- 4 to 727 +/- 109 pg/mL. In the second experiment two weeks later, epinephrine was again infused (112 +/- 17 to 727 +/- 148 pg/mL) but with somatostatin (0.8 microgram/kg/min) and intraportal replacement amounts of insulin and glucagon. The pancreatic hormones were administered in such a way as to mimic the insulin and glucagon levels observed during epinephrine infusion in the first experiment. In the first experiment, epinephrine caused changes in insulin and glucagon levels at five minutes of plus 9 +/- 1 microU/mL and minus 1 +/- 3 pg/mL, respectively, and averaged plus 6 +/- 2 microU/mL and minus 9 +/- 5 pg/mL over the first hour. Glucose production peaked at 15 minutes (increment of 0.83 +/- 0.24 mg/kg/min) and increased by an average of 0.38 +/- 0.12 mg/kg/min in the first hour. In the second experiment, intraportal replacement of insulin and glucagon during epinephrine infusion resulted in changes in insulin and glucagon levels at five minutes of plus 8 +/- 3 microU/mL and plus 2 +/- 2 pg/mL, respectively, and averaged plus 4 +/- 2 microU/mL and minus 7 +/- 6 pg/mL over the first hour.(ABSTRACT TRUNCATED AT 250 WORDS)     

An audit of consent refusals in clinical research at a tertiary care center in India

Background and Rationale:

Ensuring research participants’ autonomy is one of the core ethical obligations of researchers. This fundamental principle confers on every participant the right to refuse to take part in clinical research, and the measure of the number of consent refusals could be an important metric to evaluate the quality of the informed consent process. This audit examined consent refusals among Indian participants in clinical studies done at our center.

Materials and Methods:

The number of consent refusals and their reasons in 10 studies done at our center over a 5-year period were assessed. The studies were classified by the authors according to the type of participant (healthy vs patients), type of sponsor (investigator-initiated vs pharmaceutical industry), type of study (observational vs interventional), level of risk [based on the Indian Council of Medical Research (ICMR) “Ethical Guidelines for Biomedical Research on Human Participants”], available knowledge of the intervention being studied, and each patient''s disease condition.

Results:

The overall consent refusal rate was 21%. This rate was higher among patient participants [23.8% vs. healthy people (14.9%); P = 0.002], in interventional studies [33.6% vs observational studies (7.5%); P < 0.0001], in pharmaceutical industry-sponsored studies [34.7% vs investigator-initiated studies (7.2%); P < 0.0001], and in studies with greater risk (P < 0.0001). The most common reasons for consent refusals were multiple blood collections (28%), inability to comply with the study protocol (20%), and the risks involved (20%).

Conclusion:

Our audit suggests the adequacy and reasonable quality of the informed consent process using consent refusals as a metric.KEY WORDS: Autonomy, consent, India, reason, refusal, risk     

The friction of explanted hip prostheses

Charnley prostheses, retrieved at revision surgery, were studied to assess the effects of friction on the total hip replacement procedure. Frictional resistance was measured using the Durham hip function simulator under both dry and lubricated conditions. The friction factor values (f) for the explanted prostheses were found to have a non- Gaussian distribution with medians of 0.13 [inter-quartile range (IQR) 0.10-0.16] and 0.06 (IQR 0.005-0.08) for dry and lubricated (n = 0.01 Pa s) regimes, respectively. New Charnley prostheses had values of f equal to 0.11 +/- 0.025 and 0.04 +/- 0.01 under the same conditions, and showed no large deviation from a Gaussian distribution. There was found to be a statistically significant difference in the medians of the friction factors for new and retrieved prostheses in the lubricated regime. Ingression of cement into the worn region of the cup was found to increase the friction factor significantly under dry conditions. There was no evidence of an increase in the friction factor or torque for those joints that had a loose socket with respect to those that were fixed at revision. A decrease in the frictional torque against number of cycles undergone by the joint in vivo may indicate that a fatigue-type process may have a role in the loosening of the socket. However, this relationship was found not to be significant for friction measured under lubricated conditions and it seems unlikely that the frictional torque generated in this type of prosthesis will contribute significantly to the long-term loosening of the socket.