The absorption of ultraviolet light by cell nuclei. A technique for identifying neoplastic change.

A technique for measuring the absorption of 260-nm ultraviolet light by cell nuclei is described. The results of such measurements of normal thyroid epithelial cells and benign and malignant thyroid neoplastic cells demonstrate a progressive increase in absorbance that correlates with the histologic appearance of neoplasia. The possible theoretic basis for this phenomenon is explored. The increased nuclear absorbance observed in neoplastic cells is hypothesized to result from the disruption of hydrogen bonds between the DNA base pairs, which allows unwinding of the double helix and loss of the normal control of mitosis.

Increased peritoneal fluid lactic acid values and progressive bowel strangulation in dogs.

In the present study, we determined values for peritoneal fluid and blood ammonia, amylase, and lactic acid content in 11 dogs after experimental bowel strangulation. In the nine survivors of this group, we demonstrated a near linear increase in peritoneal fluid lactic acid over a 24-hour period, during which the plasma lactic acid remained relatively constant. A significant increase in peritoneal fluid ammonia was also demonstrated, but this did not occur until more than 16 hours after strangulation. In a second group of two dogs in which simple obstruction was produced, no significant increase in peritoneal fluid lactic acid or ammonia occurred. We did not find significant changes in peritoneal fluid amylase in this study. These data suggest that the determination of peritoneal fluid lactic acid content, especially when compared with concomitant plasma lactic acid, may be useful in the accurate preoperative diagnosis of bowel strangulation.

The affinity glycated hemoglobin in a family with hereditary spherocytosis and in other non-hemoglobinopathic hemolytic anemias.

The glycated hemoglobin (GHb) is lowered by hemolytic anemia. The cation-exchange HbA1 has been shown to be lowered by hereditary spherocytosis (HS). The HbA1, however, can be increased by elevations of fetal hemoglobin (HbF). The affinity GHb, a parameter related to, but not identical with, the HbA1, and unaffected by HbF, has been shown to be low in hemoglobinopathies but not, to our knowledge, in HS and other non-hemoglobinopathic hemolytic anemias. Therefore, the affinity GHb and HbF was determined in four members of an HS family and in nine other cases of non-hemoglobinopathic hemolytic anemia, including three autoimmune hemolytic anemias, four red cell fragmentation syndromes (two "Waring blender" syndromes, one thrombotic thrombocytopenic purpura in association with tumor, and one case of disseminated intravascular coagulation), and two red cell membrane defects: paroxysmal nocturnal hemoglobinuria and another case of hereditary spherocytosis. The GHb for these nine cases was 3.6 +/- 1.7 percent (normal 6.0 +/- 2.0 percent; p less than 0.001). The reticulocyte count, available in four cases, was 0.23 +/- 0.14 and correlated negatively with the GHb. The average GHb in the HS family was 3.9 +/- 0.8 percent, which was significantly less than the normal of 6.0 +/- 2.0 percent (p less than 0.001); the HbF was less than 1.0 percent. It is concluded that the GHb is diminished in hemolytic anemias not associated with hemoglobinopathies and that this lowering reflects the shortened red cell life span in these processes. To our knowledge, this is the first report of low GHb in hemolytic anemia not associated with hemoglobinopathy, by the affinity chromatographic technique, as opposed to the cation-exchange chromatographic technique.

Automated leukocyte differentials in trisomy 13.

We have reported a case of a 4-month-old girl who died with trisomy 13. Nuclear projections in polymorphonuclear leukocytes are diagnostically important in this syndrome and are not identified by automated leukocyte differential counting. The presence of polymorphonuclear leukocyte nuclear projections on the blood smear was identified only after the results of karyotyping were known. Recently, the importance of physician review of blood smears in anemia has been challenged. We believe this case, however, reaffirms the importance of physician blood smear review in suspected cytogenetic disease.

Densitometry and microchromatography compared for determination of the hemoglobin C and A2 proportions in hemoglobin C and hemoglobin SC disease and in hemoglobin C trait.

Using both densitometry and anion-exchange microchromatography, we measured hemoglobin C (Hb C) and Hb A2 proportions in 11 patients, eight of whom had Hb AC, two Hb SC, and one Hb CC. For one patient with Hb SC, we made the determinations before and after a transfusion. The mean (and SD) for the sum of Hb C + Hb A2 by densitometry and anion-exchange microchromatography for the nine patients with Hb AC and Hb CC were 45 (18) and 40 (18)%, respectively (p greater than 0.1, r = 0.98); for the three determinations involving the two Hb SC patients, the respective proportions were 40 (9.9) and 38 (6.6)% (r = 0.88). Electrophoretic analysis of microchromatographic eluates from the Hb AC and Hb CC patients showed that 6% of the absorbance of the late high-ionic-strength eluate was due to Hb C, which was responsible for the statistically insignificant difference between densitometric and chromatographic values for Hb C + A2 values. Electrophoresis on cellulose acetate of concentrated eluates of the Hb C + A2 fraction from the two Hb SC patients revealed no contamination by Hb S. Evidently, microchromatography can be used to determine Hb C + A2 in patients with Hb C or Hb SC disease or Hb C trait.

The microchromatographic measurement of fetal hemoglobin levels in hemoglobin C-associated conditions.

We report the microchromatographic estimation of fetal hemoglobin (Hb F) in 3 conditions associated with hemoglobin C: hemoglobin C-hereditary persistence of fetal hemoglobin (Hb C-HPFH), homozygous hemoglobin C disease, and hemoglobin SC disease. In the case of Hb C-HPFH (Case 1) the Hb F level by alkali denaturation was 32.5% while the Hb F level by microchromatography was 35.3%. In both cases of homozygous Hb C disease and Hb SC disease the Hb F level by alkali denaturation was less than 1% and the microchromatographic Hb F level was 0.7%. We conclude that microchromatographic methods for determination of Hb F levels can be employed in the Hb C-HPFH, homozygous Hb C, and Hb SC conditions. To our knowledge, the microchromatographic determination of Hb F levels has not been applied previously to the Hb C-HPFH condition or Hb C disease.

Factor VIII related antigen-concanavalin A binding in plasma and serum.

The Factor VIII Related Antigen (FVIIIRAG) was determined by electroimmunodiffusion and the per cent concanavalin A (% con A) binding of FVIIIRAG was determined by crossed affinity electrophoresis (CAIEP) in the plasma and serum of 8 hospitalized patients. The FVIIIRAG in plasma and serum was 669 and 501% of lyophilized plasma, respectively (correlation coefficient, r = 0.92). The FVIIIRAG con A-binding of plasma and serum were 44 and 16%, respectively (p less than 0.01), while the FVIIIRAG-con A binding of lyophilized plasma is 78%. We conclude that plasma has a higher content of electrophoretically slower, high con A affinity FVIIIRAG multimers than serum. The high FVIIIRAG levels of both serum and plasma and the comparatively low plasma FVIIIRAG-con A binding here may reflect the acute phase reaction.

Detection of fetal-maternal hemorrhage in a mother with sickle trait and hereditary persistence of fetal hemoglobin.

An Rh-negative term primigravida with sickle trait and hereditary persistence of fetal hemoglobin and an Rh-positive infant were evaluated postpartum for fetal-maternal hemorrhage (FMH) by the acid-elution test for fetal hemoglobin (HbF) cells. Using the acid-elution test, 100 percent of the cells contained HbF. The patient received one vial of Rh immune globulin and was discharged. This report illustrates that lack of specificity is a major problem of the acid-elution test for detection of fetal-maternal hemorrhage, due to its failure to differentiate true fetal Rh-positive cells from maternal Rh-negative HbF cells. Moreover, productivity studies suggest performance of agglutination tests such as the immune rosetting test is more convenient than performance of the acid-elution test to screen for FMH. The recently developed enzyme-linked antiglobulin test, however, may replace both of these procedures for detection of FMH.

The identification of chemical carcinogens by an absorption spectrum associated with DNA base pairs.

A simple spectrophotometric technique for identifying carcinogens is described based on the observation that many known carcinogens and/or their metabolites exhibit maximal absorption in the 260-273nm range. This absorption spectrum of carcinogens falls between that of DNA base pairs. Carcinogens in the enol form may possess electromagnetic properties which disrupt the normal hydrogen-oxygen bonds between the base pairs, inducing uncontrollable mitosis.