Cytosolic free Ca2+ in insulin secreting cells and its regulation by isolated organelles

Summary The role of Ca²⁺ in secretagogue-induced insulin release is documented not only by the measurements of⁴⁵Ca fluxes in pancreatic islets, but also, by direct monitoring of cytosolic free Ca²⁺, [Ca²⁺]_i. As demonstrated, using the fluorescent indicator quin 2, glyceraldehyde, carbamylcholine and alanine raise [Ca²⁺]_i in the insulin secreting cell line RINm5F, whereas glucose has a similar effect in pancreatic islet cells. The regulation of cellular Ca²⁺ homeostasis by organelles from a rat insulinoma, was investigated with a Ca²⁺ selective electrode. The results suggest that both the endoplasmic reticulum and the mitochondria participate in this regulation, albeit at different Ca²⁺ concentrations. By contrast, the secretory granules do not appear to be involved in the short-term regulation of [Ca²⁺]_i. Evidence is presented that inositol 1,4,5-trisphosphate, which is shown to mobilize Ca²⁺ from the endoplasmic reticulum, is acting as an intracellular mediator in the stimulation of insulin release.     

Enhanced detection in the aperture of focal attention during simple discrimination tasks

There is increasing evidence that it is possible to shift an aperture of focal attention to a position in visual space independent of fixation and that this can be done much faster than the eyes are able to move. Recently, we showed that such serial scrutiny by the aperture of focal attention is required before an observer is able to tell what a target is (for example, to know whether the orientation of a line segment is horizontal or vertical). Here we considered whether attention directed towards a specific position in the visual field for an orientation discrimination task improves performance on a simple detection task in the area to which attention is directed. We found that a small test flash could be detected when it was positioned near a peripheral line target presented briefly, if the orientation of the target had to be identified. The test flash could not be detected when presented at some distance from the same target or when another target had to be identified. This enhancement implies that even simple identification tasks such as orientation discrimination are not performed passively by the visual system.     

Cross-reactive monoclonal antibodies to alcohol dehydrogenases

Summary Three anti-horse liver alcohol dehydrogenase (HLADH) monoclonal antibodies are described. Two are specific for ADH and cross-react with class I and II enzymes from mouse, horse and Chinese hamster. They are specific for the native enzyme but do not inhibit enzyme activity except when combined at high concentration. The third antibody was isolated as a response to rabbit metallothionein. It binds metalloproteins and inhibits ADH activity.     

Comparison of different techniques for semiquantitative detection of HBV DNA by PCR

Conclusions Our results indicate that the choice of the probe for ELOSA is of major concern. In our panel we had seven sera which contained about 100 molecules/ml and further five sera which contained less than 1000 molecules/ml. Most of them were detected by the long probe but not by the short probe. When PCR for the S-or PreS-gene was included it was possible to detec all 24 HBV-positive sera (not shown) by ELOSA. The reliable lower quantification limit for the long probe is 250 molecules/ml and for the short probe 2500 molecules/ml. Surprisingly, chemiluminescence did not produce better qualitative or quantitative results. The data suggest that the usage of several replicates allows relative quantification in most cases. One possible drawback we see is the hybridization efficiency. Six of our positive samples showed great differences between the number of target molecules suggested by agarose gel electrophoreses or by hybridization (Southern blot or ELOSA). All of them contained more than 10⁶ molecules/ml. For these cases and for the samples where the short probe and the long probe gave discordant result (2 cases) we think that competitive PCR will be the method of choice, but in most cases ELOSA with the long probe gives reliable results and is highly sensitive.     

Peroxidase activity and thiocyanate accumulation in salivary glands

Summary Salivary glands with high, low, or no peroxidase activity do not differ in [S¹⁴CN^–] after the i.v. injection of KS¹⁴CN, nor do the glands differ from blood and muscle in [S¹⁴CN^–]. The content of SCN^– in a salivary gland does not mirror the gland's participation in the peroxidase-mediated antimicrobial mechanism.     

Mammalian arylsulfatases A and B: relative rates of hydrolysis of artificial substrates

Summary Rodent and bovine arylsulfatase B hydrolyze 4-methylumbelliferyl sulfate (4MUS) 10- to 30-fold more efficiently than arylsulfatase A. Therefore, 4MUS grossly underestimates arylsulfatase A activity in the presence of excess arylsulfatase B.     

Chemotaxis of human neutrophils against gravity: A new method

Summary A new method is described in which human neutrophils were made to migrate upward and against gravity. Thus, the possible effect of gravity on cell migration and consequent detachment of cell after migration have been eliminated.Supported in part by N.I.H. grants: No.HL19628-03, No. DE04898, No. CA24215 and No.RR05493).