Murine protein kinase CK2 alpha': cDNA and genomic cloning and chromosomal mapping.

Protein kinase CK2 (casein kinase II) is a heterotetrameric enzyme implicated in many essential regulatory pathways in cells. We have determined the sequence of the murine CK2 alpha' cDNA that encodes a 350-amino-acid protein that would have 99 and 98% homology with the human and chicken proteins, respectively, and is also highly homologous to murine CK2 alpha. To clarify the sequence of the 5' end of the cDNA and to elucidate the structure and regulation of the gene, we obtained a bacterial artificial chromosome clone that contains the 35-kb CK2 alpha' gene. The gene consists of 12 small exons; the 5' end, including the first exon and intron, is extremely GC rich and contains a CpG island. The putative promoter contains potential binding sites for a variety of transcriptional factors but appears to lack CCAAT- or TATA-like elements. A polymorphic dinucleotide repeat in the fifth intron allowed us to map the CK2 alpha' gene to murine Chromosome 8.

A flexible, computer-controlled video microscope capable of quantitative spatial, temporal, and spectral measurements.

A video microscope system has been constructed and tested that incorporates computer-controlled video cameras for high-resolution and low-light microscopy. The low-light camera system involves a dual microchannel plate-image intensifier capable of photon gain as high as 500 000 and a gated silicone-intensified target vidicon to achieve usable photon sensitivity with a noise equivalent signal of only 2 photons (500 nm) per pixel per second. We have compared the limitations and capabilities of this camera system with those of a high-resolution video camera and conventional photomicroscopy. Uses of the low-light camera coupled to a computer system include image acquisition of weak-light images from self-luminous specimens, fluorescence microscopy with weak exciting light, kinetic resolution of calcium-mediated events as monitored by the calcium-sensitive bioluminescence of aequorin, and spatially resolved spectroscopic measurements. Flexible use of this system in these various applications is possible because it allows operation with illumination intensities over a dynamic range of 100 000:1.

Instrumentation and techniques for analysis of hydrogen peroxide and peroxide-producing reactions involving earthworm (Diplocardia longa) bioluminescence.

Earthworm bioluminescence (species Diplocardia longa) is a sensitive tool for determination of peroxide titers in biological systems, with linearity of the standard curve for hydrogen peroxide extending from 10 nmol/L in the assay to greater than 1 mmol/L, and with sensitivity to 2 pmol of peroxide per assay. This assay's insensitivity to extraneous protein, turbidity, and solutes that are not redox active makes it particularly useful for automated or crude sample analysis and for analysis of oxidases and their substrates in coupled reactions. Examples of this latter application we discuss are analyses of glucose and putrescine oxidases and their substrates. In each case linear standard curves are obtained, covering several decades of concentration. The coupled reactions also allow in situ detection of oxidases in electrophoresis gels. We also describe a small, portable photometer system and an on-line computer-controlled instrument system that has facilitated development of these assays and will also allow automatic scanning of bioluminescence in gels. The data-collection and decision-making algorithms of this system are simple and easily adaptable for use in an inexpensive computer-based photometer system.

Propranolol effects on acute marihuana intoxication in man.

To investigate the possible interaction of a beta adrenergic blocking agent and marihuana, six healthy experienced marihuana smokers received the two drugs separately and in combination. Propranolol (120 mg per os) reduced resting HR and BP; there were no changes in performance on tasks designed to test psychomotor speed, attention, memory and learning. Marihuana (10 mg delta9-THC), administered in smoke, induced the typical subjective state ("high") with marked increases in HR, BP and conjunctival injection; it impaired performance on a learning test without significantly affecting attention. Pre-treatment with propranolol blocked effectively the cardiovascular effects of marihuana; it prevented the learning impairment and, to a lesser degree, the characteristic subjective experience.

Visceral learning in asthma.

The hypothesis of visceral learning has opened a new avenue in the search for a pathway between psychosocial stimuli and physiological changes. To apply this approach to asthma required a technique for the measurement of the airways' patency, which could be interfaced with the strategy of visceral learning. The method of forced oscillations was shown to correlate highly with whole body plethysmography. The rapid output of the instrument was used on line to control a visual reinforcement signal. Forty-six mild asthmatics, blind to the effect sought in order to minimize the role of suggestion, were tested in a series of evolving experiments. In the first series, two groups of subjects (N=15,13) were able, with this sensory feedback, to decrease (p less than 0.01) their total respiratory resistance; subsequently a smaller group of subjects (N=5), who received reinforcement signals unrelated to the state of their airways, showed no such change. Similar results were obtained in the second series of tests; in A-B-B-A order, the subjects (N=13) received either contingent or noncontingent reinforcement; furthermore, the reinforcement was available only if their lung volume was within the range observed during baseline.