Redistribution and unmasking of Annexin V binding sites in apoptotic Raji cells

The complement receptor type 2 (CR2) associates with other surface antigens and proteins and its redistribution and/or unmasking occurs through still unknown mechanism(s). The data presented demonstrate that high-density cultured CR2-positive cells undergo apoptosis and that the redistribution and unmasking of Annexin V binding sites occurs in a fashion similar to the redistribution and unmasking of CR2. Therefore, apoptotic and non apoptotic cells from the same lineage may share a similar mechanism for the exposition of neo-surface markers     

Postadhesive Technique for Archival Paraffin Sections

We present a postadhesive protocol for adhering paraffin sections of archival material to microscope slides. Appropriately posttreated sections, subsequently processed for immunohistochemistry, remained attached to the slides and were well preserved with no signs of artifacts, such as scratching and shrinkage. The immunohistochemical staining was intense and antigen-specific without nonspecific background. Specific staining intensity was equal to that produced in untreated control sections; however, the latter became partially or fully detached from the slides. The postadhesion protocol may be used with modern techniques and is recommended for reclaiming use of otherwise unsuitable paraffin sections of archival material.     

Endogenous Tumor Necrosis Factor α Unmasks the Binding Sites for N-Acetylglucosaminyl-(β1-4)-N-Acetylmuramyl-Alanyl-D-Isoglutamine on the Surface of Melanoma Cells

The surface of the melanoma BRO cells was shown to contain binding sites for N-acetylglucosaminyl-(1-4)-N-acetylmuramyl-alanyl-D-isoglutamine (GMDP). Their number (1500 ± 200 per cell) and affinity (K _d= 10 ± 1.2 nM) were determined. The occurrence of these sites was found to correlate with the ability of the melanoma cells to react in vitrowith GMDP by increasing the expression of melanoma-associated antigens (MAA). An increased number of the GMDP binding sites (5200 ± 500 per cell) was observed upon treating the melanoma BRO cells with tumor necrosis factor (TNF-). The mechanism of the TNF- action most likely involves the unmasking of GMDP binding sites, initially expressed on the cell surface, by activating the endogenous protease that hydrolyzes surface proteins, in particular, highly glycosylated LAMP-2 protein exposed on the melanoma cell surface.     

Fine structure of the band 3 protein in human red cell membranes: Freeze-fracture studies

The major red cell membrane protein, band 3, is a glycoprotein which extends across the membrane from the extracellular space into the cytoplasmic compartment. It is widely held that band 3 is a component of the intramembrane particles (IMP) which can be demonstrated by freeze-fracture electron microscopy. In this study, we find that the outer surface poles of the IMP can be seen by freeze-etching after they are unmasked by proteolysis under conditions which excise the surrounding sialopeptides from the membrane. The poles appear as distinctive projections, 30–50 Å in diameter, the “ES particles.” The ES particles remain associated with the outer surface of the membrane following cleavage of the band 3 polypeptide by chymotrypsin or pronase. This is consistent with previous biochemical studies which have shown that the 38,000-dalton outer surface segment of band 3 is intercalated in the lipid bilayer. A granulofibrillar component at the inner surface of the membrane is provisonally identified as the 40,000-dalton inner-surface domain of band 3.     

Functional Properties of Slowly Adapting Mechanoreceptors in Cat Footpad Skin

The functional properties of slowly adapting (SA) afferent fibers innervating cat footpad skin were examined. Measurements were taken of receptive field area; spontaneous activity (< 1 impulse/sec); the slope of the stimulus-response curve for steady indentations up to 2 mm in amplitude; variability of the interimpulse intervals, as measured by the coefficient of variation of time interval histograms; decay of the response to steady indentation; and sensitivity to sinusoidal vibration (most sensitive at 5-10 Hz). Where comparable tests were performed on glabrous and hairy skin SA fibers, the functional properties of those in glabrous skin more closely resembled SAI fibers than SAII fibers. Additional results from glabrous skin SA fibers suggest that it is distortion of the nerve endings rather than steady indentation or compression that leads to a brisk response. On the measures described above, there appeared to be only one functional class of SA fiber innervating the cat footpad skin.     

Spatial unmasking in the echolocating Big Brown Bat, <Emphasis Type="Italic">Eptesicus fuscus</Emphasis>

Masking affects the ability of echolocating bats to detect a target in the presence of clutter targets. It can be reduced by spatially separating the targets. Spatial unmasking was measured in a two-alternative-forced-choice detection experiment with four Big Brown Bats detecting a wire at 1 m distance. Depth dependent spatial unmasking was investigated by the bats detecting a wire with a diameter of 1.2 mm in front of a masker with a threshold distance of 11 cm behind the wire. For angle dependent spatial unmasking the masker was turned laterally, starting from its threshold position at 11 cm. With increasing masker angles the bats could detect thinner wires with diameters decreasing from 1.2 mm (target strength −36.8 dB) at 0° to 0.2 mm (target strength −63.0 dB) at 22°. Without masker, the bats detected wire diameters of 0.16 mm (target strength −66.2 dB), reached with masker positions beyond 23° (complete masking release). Analysis of the sonar signals indicated strategies in the echolocation behavior. The bats enhanced the second harmonics of their signals. This may improve the spatial separation between wire and masker due to frequency-dependent directionality increase of sound emission and echo reception.