A panel of antibodies for the immunostaining of Bouin's fixed bone marrow trephine biopsies.

AIMS: To assess a panel of antibodies on Bouin's fixed bone marrow trephine (BMT) biopsies. These biopsies are widely used in routine diagnosis of various haematological malignancies and may be the sole material available in many centres; however, information regarding the immunostaining of this material is lacking. METHODS: Biopsies were taken from 72 patients presenting with various haematological malignancies (leukaemia, 38; lymphoma, 14; multiple myeloma, 20). A panel of antibodies was assessed on Bouin's fixed BMT biopsies by the alkaline phosphatase-antialkaline phosphatase method. RESULTS: Three B (MB2, LN-2, Ki-B5) and two T cell lineage antibodies (UCHL-1, CD3-r) reliably identified lymphoid cells, while MPO-r, Leu-M1/CD15, and KP-1/CD68 recognised cells from the myeloid or histiocytic/macrophage series. Reed-Sternberg cells were stained by LN-2, Leu-M1, and CD30. Antibodies specific for plasma cells (VS38) and hairy cells (DBA.44) gave a variable pattern of staining. Among the proliferation markers, proliferative cell nuclear antigen but not Ki-67 related antibodies were effective. CONCLUSION: This study presents a panel of antibodies with reactivity not restricted to common fixatives that are also suitable for Bouin's fixed BMT biopsies.

Chloride and inorganic phosphate modulate binding of oxygen to bovine red blood cells.

The influence of Pi and Cl on the equilibrium of oxygen binding to bovine red blood cells was assessed by plotting the whole blood oxygen dissociation curve measured under standard conditions with and without added KCl and K2HPO4. Both salts shifted the oxygen dissociation curve to the right. This effect was more marked at the highest saturation levels. At a given saturation level, the anion-induced shift was linearly related to the concentration of salt added to the blood. Cl had a greater effect than Pi. The relationship between changes in Po2 at 50% hemoglobin saturation (in Torr) and concentrations of ions added (in mmol/l) was equal to 0.0515[Cl] + 0.0302[Pi] (r2 = 0.94; P < 0.001). These changes were not due to the hyperosmolality induced by salt addition, since sucrose added in place of salts was without effect on the measured parameters. The oxygen exchange fraction expressed as percentage of saturation, i.e., the difference in hemoglobin saturation when Po2 decreases from 130 to 40 Torr, was linearly correlated to added anion concentration (in mmol/l) (= 0.102[Cl] + 0.059[Pi] (r2 = 0.95; P < 0.001)). No significant interaction between the anions was observed; their effects were purely additive. This original mechanism of controlling the oxygen affinity of bovine blood may have clinical relevance: Cl and Pi could be used to increase oxygen transport efficiency in hypoxic animals.