Hyperbaric oxygen treatment of dogs has no effect on red cell deformability but causes an acute fluid shift.

Red blood cells respond to a number of perturbations, including hypoxia, with a reduction in deformability. Local hypoxia may become self-reinforcing, as hypoxic cells block capillaries preventing perfusion by oxygenated cells. Hyperbaric oxygen (HBO) is frequently used to treat conditions involving some degree of local hypoxia, but does it have a direct effect on deformability? To investigate this, 12 normal dogs received a 10 week "clinical" course of HBO: one 90 min treatment per weekday at 2.4 ATA (243 kPa), 100% O2. On Mondays and Fridays, a blood sample was drawn into EDTA, centrifuged, and the packed red blood cells resuspended in medium to a dilution of 2 x 10(6) to 5 x 10(6) cells/ml, and filtered under constant of 1.08 kPa through a precalibrated Nucleopore Hemafil Polycarbonate membrane. Filtrate was collected for one minute and weighed, and the red blood cell "incremental volume" calculated according to Engstrom (Engstrom and Ohlsson, Pediatric Res. 27:220-226, 1990). No significant change was seen in filtration rates, indicating that HBO itself neither improves nor impairs dog red blood cell deformability. Changes in other commonly measured blood parameters remained within clinical norms. An acute fluid shift out of red blood cells and into plasma was indicated.

Phosphorylation of algal centrin is rapidly responsive to changes in the external milieu.

Centrin, a calcium-sensitive contractile phosphoprotein, is the major component of the striated flagellar roots of the flagellate green alga, Tetraselmis striata. Flagellar roots contract in response to elevated calcium levels. Data presented here indicate that the level of centrin phosphorylation is rapidly responsive to changes in the cell's external environment. Centrin is dephosphorylated in response to elevated calcium or to heat shock. An increase in centrin phosphorylation accompanies pH shock or ethanol treatment. These changes are compared with flagellar excision, flagellar root contraction, and protein synthesis under the same treatments. We conclude that under certain conditions phosphorylation/dephosphorylation of centrin and extension/contraction of the flagellar root are uncoupled. Possible role(s) of centrin phosphorylation independent of force generation in the flagellar root are discussed.

Calcium-modulated contractile proteins associated with the eucaryotic centrosome.

Affinity-purified antibodies that recognize the 20,000-dalton molecular weight (20 kd) striated flagellar root protein of Tetraselmis striata have been used to identify antigenic homologs in other eucaryotic organisms of diverse evolutionary origins. Among the green algae, Tetraselmis and Chlamydomonas, and their colorless relative, Polytomella, the 20-kd homologs appear associated with basal bodies. This occurs most prominently in the form of flagellar roots of both striated and microtubule subtended types. Among cultured mammalian cells (PtK2 and primary mouse macrophage cell lines), flagellar root protein homologs appear as basal feet, pericentriolar fibrils, and pericentriolar satellites. Mammalian sperm cells also show flagellar root protein homologs associated with their basal bodies. We envisage a functional role for these fibrous calcium-sensitive contractile proteins in altering the orientation of centrioles or basal bodies with their associated MTOCs by responding to topological calcium fluxes.