Andy C. Reese

Clinical response to cold insoluble globulin replacement in a patient with sepsis and thermal injury

Cold insoluble globulin (fibronectin, alpha 2-surface binding glycoprotein) is a naturally occurring substance necessary for optimal stimulation of the reticuloendothelial system. While this globulin depends on macrophages as the effector cells for its opsonic function, as is true of both antibody and complement, it is neither part of nor dependent on these systems for its opsonic activity. A relatively simple bioassay developed at the Medical College of Georgia substantiated that cold insoluble globulin is severely depleted in sepsis. Cryoprecipitate, properly processed and stored, is an exogenous source of cold insoluble globulin. Infused into septic patients 10 units thawed at 2 degrees C and reconstituted to 250 ml with saline solution can temporarily restore cold insoluble globulin levels and enhance activity of the reticuloendothelial system. Proper current use dictates measurement of cold insoluble globulin levels in the infusate as well as levels in the patient and the clinical response to infusion. Our bioassay and a septic patients response to infusion of cold insoluble globulin are reported herein.

A competitive inhibition assay for gelatin binding fibronectin.

Abstract. A competitive inhibition assay for functional fibronectin (Fn), based on ELISA technology, is described. The assay measures Fns physiologic ability to bind to denatured collagen (gelatin). Affinity‐purified Fn inhibits the binding of alkaline phosphatase coupled Fn to gelatin‐coated wells of a microtiter plate in a concentration‐dependent manner. The assay range is 50–500 μg Fn/ml, which is suitable for the measurement of plasma Fn in both normal and opsonin deficient individuals. It is reproducible over an eightfold dilution of plasma and is resistant to interference by normal plasma proteins. The assay described is quick, quantitative, and reproducible, and satisfies the need for a measure of functional Fn activity in the clinical laboratory.

Effect of Plasma Fibronectin, Macrophages, and Glycosaminoglycans on Tumor Cell Growth

Fibronectin (Fn) synergizes with macrophages (M phi) in inducing cytostasis and cytotoxicity of neoplastic cells in culture. Since heparin enhances Fns opsonic activity in many systems, we investigated its effect on Fn-macrophage synergy in cytostasis. MCG-T14 (a spontaneous mouse mammary adenocarcinoma) cells (4 X 10(4] were added to wells both with and without C. parvum activated M phi monolayers. To these cultures were added increasing concentrations of Fn with or without heparin. Fn synergizes with both M phi S and heparin in inhibiting tumor cell growth. The combined cytostatic effect of Fn, heparin and M phi S is more than additive. In other experiments, MCG-T14 cells were pre-incubated for 2 hr with Fn, washed free of Fn, and treated as above. The results of these experiments were similar to coculture experiments, but the effect of heparin was even more pronounced. Dermatan sulfate and hyaluronic acid had a variable effect on Fn and Fn-macrophage induced cytostasis.

Effect of Fibronectin on Macrophage-Induced Tumor Cell Cytostasis

Purified fibronectin (Fn) mediated attachment of MCG-T14 cells, a mouse adenocarcinoma, to culture vessel surfaces. Concentrations of Fn less than 30 micrograms/ml enhanced the growth rate of these cells as judged by 3H-thymidine incorporation, whereas higher levels of Fn were inhibitory. Concentrations of Fn and macrophages, which had little or no effect on the growth rate of the T14 cells when added individually, mediated a 64% decrease in the rate of growth of these cells when cultured together. Free Fn was not required for this effect since target cells pretreated with Fn and then washed also were susceptible to growth inhibition by macrophages. These results indicate that Fn is able to mediate an enhancement of macrophage antitumor activity, probably by supporting the binding of target cells to the macrophage effector cells.

Academic success through quality-managed course design

Principles of quality management (which focus on defect prevention rather than defect correction) are applied to course design. These principles mandate that 1) the mission of the course be clearly stated and reviewed in context of the total program of study, 2) the objectives for the course or program of study be clearly defined and known to the students, 3) there be numerous intermediate steps leading to the objectives, 4) students be given immediate feedback about their performance on each step, and 5) corrective action be initiated immediately when a student fails to achieve an intermediate step. These principles apply to both individual courses and to an entire course of study. The authors found that a course incorporating these principles led to an increase in student performance and long-term retention of information.

Role of prostaglandins in controlling plasma fibronectin levels

Abstract Fibronectin is a normal glycoprotein component of plasma, interstitial fluid, and extracellular matrix which has binding sites for collagen, gelatin, actin, glycosaminoglycans, fibrin, Staphylococcus aureus, and some cells. Since it is a dimer, it can crosslink these substances to each other or to extracellular components of basement membrane, thereby affecting many physiological processes. The level of circulating fibronectin is markedly reduced following even moderate blunt or operative trauma, thermal injury, starvation, advanced cancers, hemorrhage, etc. Replacement therapy has been tried with some success in patients who become septic following multiple injuries. The reduction in plasma fibronectin has been attributed to several causes including consumption by binding to cell debris at the site of injury, binding to circulating cell debris and its subsequent removal by elements of the phagocytic system, and degradation by proteolytic cleavage. However, the amount of fibronectin removed from circulation raises some question about this. In this paper, we used indomethacin, ibuprofen, imadazole, and essential fatty acid deprivation to inhibit the synthesis of prostaglandins in young adult rats. Thirty minutes after ip administration of one of the inhibitors, the rats were subjected to a midline laparotomy and mild intestinal manipulation. Blood samples were taken at intervals following closure of the incision and analyzed for fibronectin. In all cases, the normal decline in plasma fibronectin seen in untreated rats was abrogated by inhibiting prostaglandin synthesis. Since imadazole specifically inhibits thromboxane A synthesis, this strongly suggests that thromboxanes directly or indirectly control the trauma-induced reduction in circulating fibronectin. This was confirmed by ip injection of thromboxane into the rats which resulted in a decline in plasma fibronectin levels.

Effect of Cold Insoluble Globulin on Carcinoma Cell Growth in vitro

The effects of cold insoluble globulin (CIG) on short-term cultures of Walker 256 carcinoma cells have been determined. Carcinoma cell proliferation in the presence of normal or heat-treated (opsonically inactive) rat serum, affinity-purified CIG, albumin, or medium alone was measured by determination of [3H]TdR incorporation into cellular DNA and by actual cell counts. The addition of serum or affinity-purified CIG significantly reduced proliferation compared with that which occurred in the presence of albumin or medium alone. The reduced proliferation caused by CIG is due to an inhibition of cell growth (cytostasis) rather than to cell death (cytotoxicity) as demonstrated by a lack of [125I]UdR release from prelabeled carcinoma cells incubated with CIG.

Role of the Reticuloendothelial System in Shock

The main function of the reticuloendothelial system (RES) is that of host defense. It is not surprising, therefore, that the RES plays a key role in shock, a condition which represents a struggle for survival by the organism in an adverse environment. It is a drama of attack and defense, a panoply of defense mechanisms designed to counter the effects of injury and preserve the vital organs.

Dimensions of academic freedom in research.

We most often think of academic freedom in terms of the subject and content of lectures and class materials. When controversies arise concerning research, the issues usually concern political correctness or allegations of bias arising either from the sponsorship of the work or from an interest by the researcher in the outcome. These controversies, however, often result in pressures that threaten the freedom of the academic researcher to pursue that research he or she thinks is most important or interesting. Threats to academic freedom in research can be grouped into three categories: 1) pressure to not work on controversial questions, 2) pressure to contribute to the colleges income by working in areas that attract large grants and contracts, and 3) restrictions on the publication of results of research due to national security classification or because of the proprietary interests of commercially sponsored research. For those who do bench research, the category about controversial research may not seem to be as personally relevant as the others. After all, when a college (in this article, any institution of higher learning from a junior college to a comprehensive university) hires someone to do research, the college pretty well knows the areas in which the work will be done and has approved it by hiring that person rather than another applicant. The researcher (any faculty member who is involved at least part time in research) tries to gather some students, postdocs, and technicians to form a research team and undertakes research in that area. Examples that may counter that impression include the canceled conference on potential genetic basis of propensity to violence (1), studies on the potential relationship between intelligence and race (2), and possible physiological and genetic components in homosexuality (3).