Binding of 17beta-estradiol by variants of alpha-fetoprotein in rat amniotic fluid.

Two variants of alpha-fetoprotein in rat amniotic fluid were separated by their different affinity for concanavalin A-Sepharose, which selectively binds alpha-D-manno-pyranosides and alpha-D-glucopyranosides. Both forms had the same mobility upon polyacrylamide gel electrophoresis. The binding of 17beta-estradiol per mg of alpha-fetoprotein, determined both immunologically and electrophoretically, was the same for both variants. These results indicate that a specific carbohydrate portion of the molecule is not necessary for steroid binding.

Rapidly progressive glomerulonephritis and monoclonal gammopathy.

A case of rapidly progressive glomerulonephritis associated with nephrotic syndrome, hematuria, and edema is reported. Monoclonal IgG-lambda was found in the serum and urine. Renal biopsy revealed diffuse proliferative glomerulonephritis with crescent formation. Immunofluorescent study revealed IgG and lambda in a focal segmental distribution. Subepithelial humps were found on electron microscopic examination. A spectacular feature of the deposits was the presence of organized linear fibrils within the humps. Similar fibrils were found in the mesangium and urinary space. Renal function deteriorated rapidly, necessitating hemodialysis in eight months. In addition to the present case, 24 cases of glomerulonephritis associated with "benign" monoclonal gammopathy reported since 1970 are reviewed, and the potential causal relationship between monoclonal gammopathy and glomerular involvement is stressed.

A putative CEA moiety is shared by the cotton-top tamarin (Saguinus oedipus) and humans.

CEA-like molecules immunologically distinct from those in humans have been described in non-human primates. These primates do not share the human predilection for colitis and subsequent development of colorectal cancer. CEA expression has not been fully evaluated in a lower-order primate, the cotton-top tamarin (Saguinus oedipus), an animal model for colitis and colorectal cancer. We found increased levels of CEA in both colonic washings and tissues of these animals using a commercially available kit, CEA AIA-PACK (Tosoh Medics, Foster City, CA). In contrast, we observed that other CEA kits failed to detect CEA in tamarins. To elucidate the nature of the CEA-like protein detected, we used the two component monoclonal antibodies used in the CEA AIA-PACK kit, and identified the reactive molecules by Western blotting. A band of approximately M(r) 50,000 was found to be common to samples from both humans and the tamarins. Minimal binding was observed with NCA antibody. We conclude that a CEA-like molecule shared by humans and tamarins may play a role in the pathogenesis of colitis and cancer in both species.

Carcinoembryonic antigen family of adhesion molecules in the cotton top tamarin (Saguinus oedipus).

Humans and the cotton top tamarin, a model for colitis and colorectal cancer, share carcinoembryonic antigen (CEA) moieties. We quantified CEA in colonic washings and extracts in both, and CEA bands were confirmed by Western blot. We compared CEA-family expression in tissues and serum in the tamarin with that of the common marmoset, which develops colitis but not cancer. CEA levels are higher in tamarin washings compared with humans, and higher than in marmosets extracts (P<0.005). CEA molecular species appear to be specific, and human CEA-family member epitopes are also found in these primates. The higher CEA levels in the tamarin may reflect the overall higher cancer prevalence.

Effect of fresh-frozen plasma resuscitation on cardiopulmonary function and serum protein flux.

The adverse effects of albumin resuscitation on coagulation activity, cardiopulmonary function, and extravascular flux of nonalbumin protein have made fresh-frozen plasma (FFP) an attractive alternate volume expander for hemorrhagic shock. This study addresses the effects of FFP on cardiopulmonary hemodynamics and protein flux. Whole blood was collected three and six weeks before shock, separated into red blood cells (PRBCs) and FFP, and stored. In 24 conditioned splenectomized dogs, resuscitation from reservoir shock of two hours' duration consisted of autologous PRBCs and balanced electrolyte solution (BES) in control dogs and PRBCs, BES, and FFP in plasma-treated dogs. Hemorrhagic shock reduced serum albumin and IgG levels in both groups. Resuscitation with FFP led to a higher cardiac output, left ventricular stroke work (LVSW), and pulmonary capillary wedge pressure (PCWP). The PCWP/LVSW ratio was comparable for both groups. Postshock day 2 serum albumin and IgG levels and lymphatic albumin and IgG concentrations were increased in plasma dogs. Therefore, FFP supplement to PRBC and BES resuscitation does not derange the PCWP/LVSW ratio or reduce intravascular nonalbumin proteins.

Multiple forms of bovine kidney alkaline phosphatase.

Partial purification of bovine kidney alkaline phosphatase produced two fractions (F1, F2). Double immunodiffusion experiments using antiserum against the pooled kidney fractions showed a complete fusion of the precipitin line of bovine kidney F2 with those of bovine kidney F1, liver, bone, and placental alkaline phosphatases. No reaction was observed between the antiserum and alkaline phosphatase from either bovine intestine or human kidney. Crossed immunoelectrophoresis of fraction F2 produced two peaks that crossed each other and stained for enzyme activity. This finding is discussed in light of recent reports of multiple antigenic forms of alkaline phosphatase form human and rabbit kidney.

Beta2-microglobulin in human colostrum and milk: effect of breast feeding and physico-chemical characterization.

A statistically significant increase in beta2-microglobulin concentration in babies' sera after birth was accompanied by a decrease in beta2-microglobulin concentration in sera of nursing and non-nursing mothers; the amount by which babies' sera concentrations increased was not correlated with the decrease in serum or milk concentrations in their mothers. These results suggest that breast feeding does not affect the concentration of beta2-microglobulin in babies' sera. Furthermore, there was no relationship between serum beta2-microglobulin concentration of mothers and their babies at either point of observation. In all instances, however, the beta2-microglobulin concentration was significantly higher in infants' sera than in mothers' sera.

IgG Antibodies Against Factor VIII in Normal Individuals.

Previously, we found noninhibitory antibody against factor VIII in a hemophilic subject. We screened sera from normal and hemophilic individuals for the presence of similar antibodies. Sera from 18 normal individuals and 17 hemophilia A subjects were tested for the presence of factor VIII antibodies. We also tested two commercial immunoglobulin preparations for the presence of similar antibodies. Serum from two normal individuals were found to have IgG1 and IgG2 antibodies against factor VIII. The two commercial immunoglobulin preparations also contained IgG1 and IgG2 antibodies against factor VIII. Conclusions: Sera of apparently normal individuals contain antibodies to factor VIII with no inhibitory qualities. Such antibodies against factor VIII appear to be a common occurrence.

The effect of steroids for shock on the immune response to tetanus toxoid.

The effect of corticosteroids on the immune response after severe injury and hemorrhagic shock was studied in 88 patients, who received an average of 13 blood transfusions for injury and hemorrhagic shock, which caused the systolic blood pressure to be below 80 torr for an average of 29 minutes. The immune response to tetanus toxoid was tested in the postoperative period. Besides the administration of blood, crystalloid solution, and plasma for coagulation factor deficiency, 42 patients also received methyl-prednisolone 1 g during additional resuscitation followed by an average of 15 mg/kg given daily for the next 2 days. The total dose of methylprednisolone averaged 3.9 g. The two groups of patients were similar for resuscitation needs and for insult. The immune response to tetanus toxoid was not significantly different between the two groups of patients. These data show that a short-term bolus of massive steroids does not appear to alter, significantly, the immune mechanism following severe hemorrhagic shock.

IgD myeloma protein with "unreactive" light chain determinants.

Serum from a patient with multiple myeloma showed a monoclonal protein, classified by immunoelectrophoresis as IgD. Immunofixation electrophoresis and immunoelectrophoresis failed to demonstrate a precipitation reaction between the paraprotein and antisera to immunoglobulin light chains. The light chains of the monoclonal protein, immunologically inaccessible in the intact molecule, reacted with anti-lambda chain antisera only after reduction and alkylation of the paraprotein. Moreover, interpretation of the immunoelectrophoretic patterns was hampered by the presence in patient's serum of free lambda chains having about the same mobility as that of the paraprotein.

A new approach to immunologic identification of factor VIII antibodies.

We tested for antibodies against factor VIII by using monoclonal antibody-purified factor VIII preparation as a source of antigen. The factor VIII was adsorbed on nitrocellulose membranes and stored in a refrigerator until later use. Plasma or serum was incubated with the factor VIII containing strip and the antibody was detected by another incubation with peroxidase-labelled antihuman immunoglobulin antibodies. The test was efficient in detecting antibodies in haemophilic and normal subjects with acquired inhibitors to factor VIII. It also detected antibodies to the factor VIII protein in a haemophilic subject with no evidence of inhibitor. The technique is simple, readily applicable, and serves as a useful screening tool for detecting factor VIII antibodies. The stability of the antigen-containing strips in a refrigerator is a practical advantage with potential commercial application.

Characterization of glomerular permeability and proteinuria in acute hypertension in the rat.

The effects of acute angiotensin II (AII) induced hypertension on renal hemodynamics, urinary excretory rates, and clearances of endogenous proteins, together with colloidal iron staining and numerical density of differently charged ferritins in glomerular basement membrane (GBM) have been studied. AII decreases para-aminohippurate clearance (63%) more than glomerular filtration rate (GFR) (42%), resulting in an increased filtration fraction (54%). Simultaneously, large increments in the excretory rates and clearances of albumin and IgG2a occur. The number of native ferritin particles per unit volume of GBM and its different layers increases significantly in both superficial and juxtamedullary glomeruli as a result of acute hypertension. In contrast, the number of cationized ferritin particles per unit volume of GBM as well as colloidal iron staining of GBM and adjacent cell membranes remain unchanged, irrespective of AII treatment. The results demonstrate that acute AII-induced hypertension enhances glomerular permeability to proteins of different size and shape in the absence of detectable alterations in the fixed negative charges of the GBM. Since both RBF and GFR are decreased, the increased transglomerular passage of proteins in acute hypertension appears to be due to an increase in the pore size of the glomerular filter, induced possibly by either high intracapillary pressure and/or a direct action of AII on GBM constituents.

Gastrointestinal tract antigenic profile of cotton-top tamarin, Saguinus oedipus, is similar to that of humans with inflammatory bowel disease.

As an animal model for human inflammatory bowel disease and colorectal cancer, the cotton-top tamarin remains controversial. Demonstration of antigenic similarity to the human would enhance its validity. Using colonic extracts and washings, we compared binding of seven monoclonal antibodies reactive with bowel and cancer antigens in both tamarins and humans with inflammatory bowel disease. Additionally, telomerase activity was tested for. Expression of a mucin antigen specific to human cancer was increased in tamarin colonic washings as well as aminoproteoglycans and EGFR in tamarin extracts, as compared to those of humans with inflammatory bowel disease (P < 0.005). An adenoma-associated antigen and k-ras p21 protein were negative in the tamarin. A trend to greater telomerase activity exists in tamarins. The antigenic similarity validates this model for human inflammatory bowel disease and colorectal cancer. A trend to increased telomerase activity in tamarins is consistent with the greater predisposition to cancer in these animals.

Glomerular permeability to endogenous proteins in the rat: effects of acute hypertension.

The distribution of endogenous albumin and immunoglobulin G (IgG) has been studied by the immunoperoxidase technique in the superficial glomeruli of both normotensive and hypertensive (acute angiotensin II-induced) Munich-Wistar rats. Endogenous IgG has also been detected in rats immunized with horseradish peroxidase. Labeled antibodies have been applied to sections on a conventional manner as well as by an electrophoretic technique. The immunization of animals with horseradish peroxidase, as well as application of the electrophoretic technique, both result in a greater yield of labeled glomeruli. Albumin is present within the capillary lumina of control animals, penetrates the capillary walls, and extends into the urinary space. Endogenous IgG is mainly confined to lumina of glomerular capillaries, with only small amounts visible in the laminae rarae of the basement membrane. After acute hypertension induced by angiotensin II, there is increased staining of albumin and IgG in the basement membrane and of albumin in the urinary space. There is also penetration of IgG into Bowman's space. Both macromolecules are found in dilated mesangial channels. These modifications of glomerular permselectivity in hypertension are not accompanied by discernible ultrastructural changes in the peripheral capillary wall. It is suggested that the transcapillary passage of albumin and IgG is dependent upon hemodynamic factors and/or subtle changes in the filtering membrane.