Optical biosensor assay (OBA).

We describe a new biosensor immunoassay involving optical diffraction to detect clinically important analytes in human body fluids. A silicon wafer is used as a support for immobilization of antigen or antibody. The protein-coated surface is illuminated through a photo mask to create distinct periodic areas of active and inactive protein.

Immunocytochemical localization of phosphatase inhibitor-1 in rat brain.

The localization of phosphatase inhibitor-1 was investigated in rat brain by use of immunocytochemistry. Studies were performed with an affinity purified IgG raised against purified rabbit skeletal muscle inhibitor-1. In rat brain tissue homogenates, this antibody reacted only with a 29 kDa protein corresponding to inhibitor-1.

Characterization in mammalian brain of a DARPP-32 serine kinase identical to casein kinase II.

DARPP-32, a dopamine- and cyclic AMP-regulated phosphoprotein of Mr 32,000, is phosphorylated in vitro by casein kinase II at a site which is also phosphorylated in intact cells. In the present study, we show that a protein kinase activity, present in caudate-putamen cytosol, phosphorylates DARPP-32 on a seryl residue located on the same thermolytic peptide that is phosphorylated by purified casein kinase II. This DARPP-32 serine kinase was indistinguishable from casein kinase II on the basis of a number of biochemical criteria.

Immunocytochemical localization of the neuron-specific form of the c-src gene product, pp60c-src(+), in rat brain.

Neurons express high levels of a variant form of the c-src gene product, denoted pp60c-src(+), which contains a 6 amino acid insert in the amino-terminal half of the c-src protein. We have determined the localization of pp60c-src(+) in neurons using an affinity-purified anti-peptide antibody, referred to as affi-SB12, that exclusively recognizes this neuron-specific form of the c-src gene product. Using affi-SB12, we examined the distribution of pp60c-src(+) by immunoperoxidase staining of sections through adult rat brains, pp60c-src(+) was widely distributed in rat brain and appeared to be differentially expressed in subpopulations of neurons.

Activation of multiple genes by provirus integration in the Mlvi-4 locus in T-cell lymphomas induced by Moloney murine leukemia virus.

Moloney murine leukemia virus-induced rat T-cell lymphomas harbor proviruses integrated near c-myc and near Mlvi-1/Mis-1/Pvt-1, another locus of common integration which maps 270 kilobases 3' of c-myc. In this report, we present the characterization of a new locus of common integration in Moloney murine leukemia virus-induced T-cell lymphomas (Mlvi-4) which maps 30 kilobases 3' of c-myc, between c-myc and Mlvi-1. The Mlvi-4 locus, whose chromosomal map location is conserved in rats, mice, and humans, is also the target of chromosomal rearrangements in a variety of animal and human tumors.

Differential expression of ARPP-16 and ARPP-19, two highly related cAMP-regulated phosphoproteins, one of which is specifically associated with dopamine-innervated brain regions.

ARPP-16 and ARPP-19 are 2 cAMP-regulated phosphoproteins of Mr = 16,000 and 19,000, respectively, which are identical except for the presence of 15 additional amino acids on the NH2-terminus of ARPP-19. The phosphorylation of these 2 proteins is regulated by cAMP and vasoactive intestinal peptide in reaggregate striatal cultures (Girault et al., 1988).

Active frequency stabilization of a 1.062-microm, Nd:GGG, diode-laser-pumped nonplanar ring oscillator to less than 3 Hz of relative linewidth.

We present results on the frequency stabilization of two diode-laser-pumped ring lasers that are independently locked to the same high-finesse interferometer. We measure the relative frequency stability by locking the lasers one free spectral range apart and observing the heterodyne beat note. The resultant beat note width of 2.

Localization of DARPP-32 immunoreactive neurons in the bed nucleus of the stria terminalis and central nucleus of the amygdala: co-distribution with axons containing tyrosine hydroxylase, vasoactive intestinal polypeptide, and calcitonin gene-related peptide.

The distribution and morphology of neurons containing the dopamine- and cyclic AMP-regulated phosphoprotein, DARPP-32, were investigated in the bed nucleus of the stria terminalis (BST) and the central nucleus of the amygdala (CeA). DARPP-32 immunoreactive neurons are numerous in both regions, but are restricted to the lateral dorsal and the lateral juxtacapsular subdivisions of the BST, and the central lateral and lateral capsular subdivisions of the CeA. Immunoreactive neurons in the lateral dorsal BST, and the central lateral and lateral capsular CeA are similar morphologically, while those in the juxtacapsular BST appear to be a subpopulation of striatal medium-sized spiny neurons.

Injection locking of a 13-W cw Nd:YAG ring laser.

A lamp-pumped, 13-W cw Nd:YAG ring laser at 1.064 microm is injection locked using a 40-mW single-frequency diodelaser-pumped Nd:YAG laser as the master oscillator. The phase fidelity of the injected slave to the master is measured using an all-optical technique.

Spatial relationship of the striatonigral and mesostriatal pathways: double-label immunocytochemistry for DARPP-32 and tyrosine hydroxylase.

Antibodies to tyrosine hydroxylase and DARPP-32 were used to examine the spatial arrangement between mesostriatal dopamine projections and the reciprocal pathway from DARPP-32-containing neurons in the basal forebrain. Use of a double-labeling immunocytochemical procedure demonstrated that the mesostriatal and striatonigral pathways run in close proximity throughout the rostral mesencephalon and basal forebrain. The majority of descending axons immunoreactive for DARPP-32 appear to originate in the striatum, including the nucleus accumbens, and run through the internal capsule to innervate the globus pallidus, entopeduncular nucleus, and all subdivisions of the substantia nigra.

Human neutrophils contain and bind high molecular weight kininogen.

Because plasma kallikrein activates human neutrophils, and in plasma prekallikrein (PK) circulates complexed with high molecular weight kininogen (HMWK), we determined whether HMWK could mediate kallikrein's association with neutrophils. HMWK antigen (237 +/- 61 ng HMWK/10(8) neutrophils) was present in lysates of washed human neutrophils. Little if any plasma HMWK was tightly bound and nonexchangeable with the neutrophil surface.

High molecular weight kininogen inhibits fibrinogen binding to cytoadhesins of neutrophils and platelets.

Fibrinogen inhibited 125I-high molecular weight kininogen (HMWK) binding and displaced bound 125I-HMWK from neutrophils. Studies were performed to determine whether fibrinogen could bind to human neutrophils and to describe the HMWK-fibrinogen interaction on cellular surfaces. At 4 degrees C, the binding of 125I-fibrinogen to neutrophils reached a plateau by 30 min and did not decrease.

The distribution of dopamine-beta-hydroxylase, neuropeptide Y and galanin in locus coeruleus neurons.

The locus coeruleus (LC) is composed of noradrenaline-producing neurons that project widely throughout the neuraxis. Subpopulations of LC neuron perikarya have been shown to contain neuropeptide Y (NPY) and galanin (GAL). In the major terminal fields of LC projections, the cerebral cortex, dorsal thalamus and cerebellar cortex, there are differing plexuses of dopamine-beta-hydroxylase (DBH), NPY and GAL immunoreactive axons.

Efficient monolithic MgO:LiNbO(3) singly resonant optical parametric oscillator.

We have operated a monolithic MgO:LiNbO(3) singly resonant optical parametric oscillator (OPO) as both a standing-wave and a ring-geometry resonator. The OPO was pumped by the second harmonic of an amplified single-mode diode-laser-pumped Nd:YAG laser. Pump depletions of greater than 60% were observed when pumping four times greater than the 35-W threshold.

Noradrenaline and neuropeptide Y innervation of the rat hypothalamus are differentially affected by 6-hydroxydopamine.

The neuropeptide Y (NPY) innervation of the hypothalamus is thought to arise largely from noradrenaline (NA) neurons of the medullary tegmentum in the rat. This view was tested in this study by analyzing the effects of intraventricular injections of 6-hydroxydopamine (6-OHDA) on hypothalamic NA and NPY innervation. 6-OHDA markedly depletes or eliminates the NA innervation of the hypothalamus, as demonstrated by dopamine-beta-hydroxylase immunohistochemistry, but does not affect the NPY innervation of the hypothalamus.

Noradrenaline neuron plasticity in developing rat brain: effects of neonatal 6-hydroxydopamine demonstrated by dopamine-beta-hydroxylase immunocytochemistry.

The present study was conducted to assess the morphological changes produced by neonatal administration of 6-hydroxydopamine (6-OHDA) in the noradrenergic innervation of the developing and adult rat brain. As demonstrated by dopamine-beta-hydroxylase (DBH) immunohistochemistry, the major alterations are the following. First, neocortical and hippocampal noradrenergic innervation is permanently eliminated by the treatment, with lesser effects on other telencephalic structures.

Inhibition of thrombin-induced platelet aggregation by high molecular weight kininogen.

1. Thrombin-induced platelet aggregation and cleavage of [3H]FSBA-labeled aggregin is mediated by platelet calpain. 2.

Neuropeptide Y localization in the rat amygdaloid complex.

Neuropeptide Y (NPY)-, avian pancreatic polypeptide (APP)-, and molluscan cardioexcitatory peptide (FMRF)-like immunoreactivity in the amygdaloid complex of the rat was investigated immunohistochemically. The distribution of each of these peptides within the amygdala is identical and cross-blocking studies indicate that all three antisera recognize the NPY antigen. Morphologically distinct populations of NPY immunoreactive neurons are differentially distributed in the medial amygdaloid nucleus and at the base of the stria terminalis.

High molecular weight kininogen binds to unstimulated platelets.

Studies were performed to determine if the unstimulated platelet membrane has a site for high molecular weight kininogen (HMWK) binding. 125I-HMWK bound to unstimulated platelets. Zn++ was required for 125I-HMWK binding to unstimulated platelets and binding was maximal at 50 microM Zn++.

Plasma prekallikrein assay: reversible inhibition of C-1 inhibitor by chloroform and its use in measuring prekallikrein in different mammalian species.

The assay of plasma prekallikrein requires activation of prekallikrein to kallikrein and sufficient inactivation of the plasma protease inhibitors of kallikrein to accurately measure the generated kallikrein activity. One method of elimination of the plasma protease inhibitors to kallikrein is to chemically pretreat the plasma. Methylamine has previously been employed to selectively inactivate alpha 2-macroglobulin.

Coherent anti-Stokes Raman scattering from small volumes.

We have used coherent anti-Stokes Raman scattering spectroscopy to resolve the Raman Q-branch line [Q (2) at 2987.18 cm(-1)] of dueterium gas contained in a 60-microm-diameter glass sphere. The 60-microm spheres contained only 10(+13) molecules at standard temperature and pressure; this made it possible to study gases that in larger quantities would be too dangerous or expensive to use.