Anti-IgM induces transforming growth factor-beta sensitivity in a human B-lymphoma cell line: inhibition of growth is associated with a downregulation of mutant p53

We wished to examine the role of transforming growth factor-beta (TGF- beta) in the regulation of human lymphoma cell growth. The RL cell line is an immunoglobulin M (IgM)+, IgD+ B lymphoma cell line, which does not constitutively express receptors for TGF-beta, and thus has lost the ability to respond to the inhibitory effects of TGF-beta. We demonstrate here that anti-Ig antibodies can efficiently upregulate the expression of TGF-beta receptors and promote sensitivity to growth inhibition by TGF-beta. Furthermore, because TGF-beta has been shown to function in late G1 of the cell cycle, we examined the ability of TGF- beta to modulate two tumor suppressor proteins known to be critical regulators of the G1/S transition, Rb and p53. Rb is a 105- to 110-kD phosphoprotein, which has been shown to maintain its growth suppressive function when it is found in the hypophosphorylated state. Wild-type p53 is a 53-kD phosphoprotein that appears to be important in preventing cell-cycle progression and promoting apoptosis in cells with DNA damage, whereas mutant p53 can overcome those functions. We show here that TGF-beta treatment of phorbol myristate acetate (PMA) or anti- Ig-activated RL cells results in growth inhibition through a dual effect on Rb and mutant p53. After TGF-beta treatment, we observe a predominance of Rb in the hypophosphorylated, growth suppressive form. In addition, we show a decrease in levels of mRNA and protein for mutant p53. We also show that, although these changes are sufficient to halt progression through the cell cycle, the cells do not appear to undergo extensive programmed cell death following 72 hours of TGF-beta treatment. Thus, although these lymphoma cells maintain the capacity to be negatively growth regulated by TGF-beta, the ability of TGF-beta to induce apoptosis must be independently controlled.     

Diverging associations of an intended early invasive strategy compared with actual revascularization, and outcome in patients with non-ST-segment elevation acute coronary syndrome: the problem of treatment selection bias

AIMS: In several observational studies, revascularization is associated with substantial reduction in mortality in patients with non-ST-segment elevation acute coronary syndrome (nSTE-ACS). This has strengthened the belief that routine early angiography would lead to a reduction in mortality. We investigated the association between actual in-hospital revascularization and long-term outcome in patients with nSTE-ACS included in the ICTUS trial. METHODS AND RESULTS: The study population of the present analysis consists of ICTUS participants who were discharged alive after initial hospitalization. The ICTUS trial was a randomized, controlled trial in which 1200 patients were randomized to an early invasive or selective invasive strategy. The endpoints were death from hospital discharge until 4 year follow-up and death or spontaneous myocardial infarction (MI) until 3 years. Among 1189 patients discharged alive, 691 (58%) underwent revascularization during initial hospitalization. In multivariable Cox regression analyses, in-hospital revascularization was independently associated with a reduction in 4 year mortality and 3 year event rate of death or spontaneous MI: hazard ratio (HR) 0.59 [95% confidence interval (CI) 0.37-0.96] and 0.46 (95% CI 0.31-0.68). However, when intention-to-treat analysis was performed, no differences in cumulative event rates were observed between the early invasive and selective invasive strategies: HR 1.10 (95% CI 0.70-1.74) for death and 1.27 (95% CI 0.88-1.85) for death or spontaneous MI. CONCLUSION: The ICTUS trial did not show that an early invasive strategy resulted in a better outcome than a selective invasive strategy in patients with nSTE-ACS. However, similar to retrospective analyses from observational studies, actual revascularization was associated with lower mortality and fewer MI. Whether an early invasive strategy leads to a better outcome than a selective invasive strategy cannot be inferred from the observation that revascularized patients have a better prognosis in non-randomized studies.     

Application of a sol particle immunoassay to the determination of affinity constants of monoclonal antibodies

The affinity constants (Ka) of monoclonal antibodies (MAb) for binding to their corresponding antigens (Ag), unlabelled and in buffered solution were determined by the following procedure: 1. Incubation of MAb (fixed concentration) with Ag (concentration dilution series). 2. Rapid bound/free separation by adding immobilized second antibody, followed by centrifugation. 3. Determination of free Ag in the supernatant using a gold sol particle agglutination immunoassay (SPIA) in a microtitration plate format. 4. Calculations and interpretation were based on Scatchard and Sips plots. Ka values found by this procedure were found to be similar to those obtained by a radio-immunoassay (RIA) procedure. The present method avoids possible artefacts in Ka values introduced by the procedure or chemical modification due to labelling of MAb or Ag. It enables rapid, simultaneous screening of a considerable number of different MAbs under non-specialized (i.e. RIA) laboratory conditions.     

Moloney cell surface antigen (MCSA) has properties of an env gene product that is serologically distinct from the env gene products of the moloney strain of murine leukemia virus

Moloney cell surface antigen (MCSA) is serologically detectable on Moloney murine leukemia virus (Mo-MuLV)-induced lymphomas. It is probably related to the tumor-associated transplantation antigen of these lymphomas. We characterized MCSA by using the typing antiserum in radioimmunoprecipitations followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Surface-iodinated Mo-MuLV-infected cells were used to identify the molecular nature of MCSA. It appeared that MCSA was expressed as an entity of 82,000 apparent molecular weight (p82^MCSA). Translation of size-fractionated mRNAs from infected cells in oocytes of Xenopus laevis showed that p82^MCSA was made on an mRNA of the same size (22 S) as that coding for the Mo-MuLV env proteins. Moreover, p82^MCSA was processed in oocytes to proteins exactly comigrating in the gel with gp70, p15E, and p12E. Competition experiments with unlabeled virus showed that MCSA and the MCSA-related env-like proteins made in oocytes were not related to any of the Mo-MuLV structural proteins. We concluded that MCSA has the properties of the product of the env gene of a murine C-type virus different from Mo-MuLV.     

Autologous bone versus calcium-phosphate ceramics in treatment of experimental bone defects

Autologous bone grafting is currently considered the treatment of choice for correction of large bone defects. However, to avoid morbidity associated with autologous bone harvesting many artificial bone-substitute materials have been developed over the years. A new generation of resorbable materials is emerging, with promising results so far. In order to investigate the possibility to use one of these new materials as an alternative with better results than hydroxyapatite, an experimental study was performed. A new resorbable calcium phosphate particles and paste forms, the latter of which hardens in situ after application. In 28 sheep, a 3-cm segmental tibial defect was made and intramedullary fixed by an interlocking nail. Twelve weeks after defect filling, radiological, biomechanical, and histological examinations were performed. Mean radiographic and biomechanical tests results were compared with the Mann-Whitney test. Significance was set at p<0.05. Radiographically, the resorbable paste group performed better than all other groups. Biomechanical investigations showed a higher torsional stiffness (p=0.049) for the resorbable calcium-phosphate paste group in comparison with autologous bone. On histological examination, no adverse effects were observed in the calcium-phosphate groups. Resorption by osteoclasts was seen in the resorbable implants. In conclusion, the current study shows an advantageous radiological and mechanical outcome for resorbable calcium phosphates. This indicates that these new materials might be a potential alternative for autologous bone grafting in humans.     

A longitudinal study of otitis media with effusion among 2- to 5-year-old African-American children in child care

OBJECTIVE: To prospectively document the prevalence of otitis media with effusion (OME) in 86 African-American children between ages 2 and 5 years. STUDY DESIGN: Eighty-six children in center-based child care whose ear status had been followed from infancy continued to be observed. Middle ear status was assessed by pneumatic otoscopy and tympanometry biweekly. RESULTS: The prevalence of OME decreased as children became older. The mean proportion of examinations demonstrating bilateral OME (BOME) ranged from 12% between 24 to 30 months to 4% between 54 to 60 months of age. The mean proportion of exams revealing bilateral normal ears increased from 77% at 24 to 30 months to 88% at 54 to 60 months of age. Although 60 children had experienced BOME that lasted 4 months or longer in the 6- to 24-month age period, only 8 of these children experienced at least 4 months of continuous BOME between 24 to 60 months. CONCLUSIONS: The proportion of time with BOME decreased progressively with increasing age in this population. Only 8 of 60 children who had experienced more than 4 consecutive months of BOME before 2 years of age continued to manifest persistent effusion or experience recurrences of prolonged BOME after 2 years of age.