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The purpose of this work is to demonstrate equivalent bipolar transistor damage response to neutrons and silicon ions. We report on irradiation tests performed at the White Sands Missile Range Fast Burst Reactor, the Sandia National Laboratories (SNL) Annular Core Research Reactor, the SNL SPHINX accelerator, and the SNL Ion Beam Laboratory using commercial silicon npn bipolar junction transistors (BJTs) and III–V Npn heterojunction bipolar transistors (HBTs). Late time and early time gain metrics as well as defect spectra measurements are reported.

Procathepsin L, the precursor to a powerful lysosomal cysteine proteinase, has been purified to apparent homogeneity from guinea pig spermatozoa, a novel and previously unrecognized source of this catalytically active zymogen. In the range of pH 5.0, procathepsin L (39,000 M r ) readily self-processed yielding a mature, single-chain proteinase (29,000 M r ) and an intact propeptide (10,000 M r ) by what appeared kinetically to be an intramolecular reaction mechanism. These characteristics resembled those reported for the “major excreted protein” (MEP) of malignantly transformed mouse fibroblasts—a protein that has been characterized as the precursor to the mouse analog of human cathepsin L (B. R. Troen, S. Gal, and M. M. Gottesman (1987) Biochem. J. 246, 731–735). Other characteristics shared by the guinea pig and mouse zymogens included proteolytic activity at pH 5.0, homologous N-terminal amino acid sequences, and immunological relatedness. It was thus concluded that acrosomal procathepsin L is the guinea pig analog of MEP. Acrosomal procathepsin L had a specific activity on benzyloxycarbonyl-Phe-Arg-7-(4-methyl)coumarylamide (Z-Phe-Arg-NMec) of 30 μmol min −1 mg −1 enzyme at pH 3.2 and 37°C. Relative to the assay substrate, rates on other fluorogenic substrates were 90% for Z-Phe-Cit-NMec, 63% for Z-Leu-Leu-Arg-NMec, 43% for D-Phe-Ser(Bzl)-Phe-Phe-Ala-Ala-p-aminobenzoate (a “specific” cathepsin D assay substrate), and 32% for Z-Val-Val-Arg-NMec. No action was detected on Z-Arg-Arg-NMec or Arg-NMec. Mature cathepsin L showed the same relative order of substrate specificity as its proenzyme form, but the absolute rates were about 5-fold greater. Additionally, the mature (single-chain) form of cathep sin L displayed K m and k cat values on Z-Phe-Arg-NMec that yielded an exceptionally high catalytic coefficient (11,600 s −1 m M −1 ) compared to values reported for two-chain forms of cathepsin L. Self-processing by acrosomal procathepsin L at pH 5.5 was totally inhibited by leupeptin, cystatin C, Ep-475, and Z-Phe-Phe-CHN 2 at 1 μ M levels. Gossypol (0.1 m M ) gave 94% inhibition. Interestingly, dextran sulfate (100 μg ml −1 ) gave a 3.6-fold increase in the rate of self-processing seen at pH 5.5—a phenomenon of potential physiological relevance in view of the high-negative-charge density present within the hyaluronic acid-rich outer layer (cumulus oophorus) of the ovum.

The morphology and chemical structure of Parylene F and Parylene N films annealed in air, N2, and vacuum were examined. The decomposition temperatures of Parylene N in air, N2, and vacuum were determined to be 175,350, and 425°C, respectively. The decomposition temperatures of Parylene F in air and N2 were determined to be 400 and 500°C, respectively. For both materials, the decomposition process with and without the presence of O is different. In the case of Parylene N, O diffuses into the materials and reacts with the C in the polymer. In the case of Parylene F, no reaction products with O are observed on the surface. Annealing at the decomposition temperature resulted in the decomposition of -CF2- functional groups to -CF- functional groups in the Parylene F. Thicker films,< μ, of either material cracked during annealing while films

Photochemical reactivity has been observed in solution-phase azulene. Experiments are described which confirm photoinduced deuteration, chlorination, and polymerization. The threshold for the observed chemistry is at the origin of S 2 which is at 27956±8 cm -1 (357.7±0.1 nm) in chloroform. Only one photon is required to induce the observed chlorine substitution reactions. Although no naphthalene is formed, this chemical channel appears to be the equivalent of a thermal reaction, with the substitutions taking place at the two equivalent positions on the small ring of azulene. The mechanism of these reactions is bimolecular in nature

Antibodies have been raised against synthetic peptides derived from the predicted primary sequence of the human cocaine- and antidepressant-sensitive norepinephrine (NE) transporter (NET). One antibody (N430), raised and purified against a putative intracellular human norepinephrine transporter (hNET) epitope, detects hNET expression in a stably transfected cell line (LLC-NET) by indirect immunofluorescence only in the presence of detergent, while no immunoreactivity is observed in either the parental cells (LLC-PK1) or in LLC-NET cells incubated with preimmune sera or peptide absorbed antibody. N430 immunoblots of LLC-NET cell extracts reveal two major immunoreactive hNET species in these cells, migrating at 80 and 54 kDa, respectively. Pulse-chase N430 immunoprecipitation studies confirm that the 54-kDa species is a transient, glycosylated intermediate of a longer lived, more highly glycosylated protein with an apparent M(r) of 80,000. In contrast, a 54-kDa species is the primary hNET product in vaccinia virus T7-infected HeLa cells, transiently transfected with hNET cDNA. PNGase F digestion of extracts prepared from LLC-NET- and hNET-transfected HeLa cells convert all immunoreactive species to a 46-kDa form, equivalent to that observed following incubation of whole cells with the glycosylation inhibitor tunicamycin. As transiently transfected HeLa and stable LLC-NET cells exhibit a pharmacologically similar NE transport activity, it appears likely that the additional glycosylation evident in the stable line does not contribute significantly to antagonist sensitivity. On the other hand, NE transport and antagonist ([125I]RTI-55) binding assays on whole LLC-NET cells treated with tunicamycin reveal a pronounced reduction in NE transport activity and hNET membrane density paralleled by an inability of NET proteins to replenish the higher M(r) hNET pool. These findings suggest an obligate role for N-linked glycosylation in hNET biosynthetic maturation, stability, and functional expression. In summary, N430 antibody is a useful tool for the visualization and characterization of hNET gene products and has permitted the first direct evaluation of biosynthetic steps leading to functional catecholamine transporter expression.

In addition to its procoagulant properties, the serine protease thrombin increases endothelial permeability, stimulates granulocyte adherence, and serves as a fibroblast mitogen. We demonstrate that thrombin is mitogenic for human lung fibroblasts in vitro. The mitogenic effect of thrombin is associated with an increase in the expression of the ligand PDGF-AA and up-regulation of PDGF alpha-receptor. Since scleroderma (systemic sclerosis; SSc) is characterized by widespread microvascular injury and is frequently complicated by pulmonary fibrosis, we sought to determine the level of thrombin activity in bronchoalveolar lavage (BAL) fluid from SSc patients and normal controls. We report a significantly higher level of thrombin activity in BAL fluid from SSc patients compared with normal controls (P0.001). Taken together, the high levels of thrombin in BAL fluid and its demonstrated mitogenicity for lung fibroblasts suggest an important role for thrombin in the pathogenesis of SSc and perhaps other fibrotic lung diseases.

In a previous study, we have demonstrated that infusion of neuropeptide Y (NPY) into the stalk-median eminence (S-ME) of gonadectomized rhesus monkeys stimulated LHRH in a dose-dependent manner. This finding led us to address the following questions: 1) What are the characteristics of NPY release in vivo? 2) How does NPY release relate to LHRH release? 3) Is endogenous NPY essential to pulsatile LHRH release? To answer these questions, three experiments using push-pull perfusion were performed in adult gonadectomized rhesus monkeys. Perfusate samples from the S-ME were collected at 10-min intervals for 6 to 12-h periods, and the concentrations of LHRH and NPY in perfusates were determined by RIA. In Exp I, the release pattern of NPY and LHRH in the S-ME was independently determined in a group of 11 conscious monkeys: NPY release in the S-ME was pulsatile with an interpulse interval of 44.9 +/- 3.3 min (n = 11). This interpulse interval was similar to that seen for LHRH release (43.8 +/- 1.1 min, n = 7). Exp II was designed to determine whether NPY pulses and LHRH pulses occur synchronously and to examine whether NPY release in the S-ME is correlated with circulating LH pulses. NPY and LHRH concentrations in aliquots of the same perfusate sample from the S-ME and circulating LH levels were concurrently measured in 8 monkeys sedated with Saffan. It was found that NPY pulses were temporally correlated (P less than 0.001) with LHRH pulses, which were also temporally correlated (P less than 0.001) with LH pulses. Moreover, NPY pulses were correlated (P less than 0.05) with LH pulses. NPY peaks preceded LHRH peaks by 4.5 +/- 0.6 min, LHRH peaks preceded LH peaks by 5.5 +/- 0.6 min, and NPY peaks preceded LH peaks by 9.7 +/- 0.8 min. In Exp III, the role of endogenous NPY in LHRH release was evaluated by infusing a specific antiserum to NPY into the S-ME during push-pull perfusion in 8 conscious monkeys. Infusion of a specific antiserum to NPY into the S-ME at 1:100 and 1:1000 dilutions suppressed pulsatile LHRH release significantly (P less than 0.05). Infusion of nonimmune serum as a control was without effect. These results are summarized as follows: 1) NPY release in the S-ME is pulsatile, 2) NPY pulses occur synchronously with LHRH and LH pulses, and 3) immunoneutralization of endogenous NPY in the S-ME suppresses pulsatile LHRH release.(ABSTRACT TRUNCATED AT 400 WORDS)

A survey of the degenerate four-wave mixing measurements and their interpretation done on composite materials composed of nanometer coated particles, such as gold-on-aluminum, metal-on-metal, gold-on-silica, and silver-on-silica, will be presented. These data show an anomalous change in slope of the phase conjugate signal versus laser intensity from 3 to 4, depending on the thickness of the coating. To understand this feature, we study the effects of the surface properties coming from the inner surface of the particle. The effects of the second harmonic generation from such an interface, as studied, for example by Shen,1 are investigated. The metallic part of the grain is modeled via a Kerr type nonlinearity in the dielectric constant. We obtain an expression for the dielectric constant in the effective medium theory. Using a Maxwell-Garnett and self-consistent field approximation, we then calculate the phase conjugate signal versus laser intensity to study the effect observed above.

L'irradiation (40 torr et 100 u/cm 2 ) de l'hexadiene-1,5 conduit aux methane, ethylene, acetylene, propene, propadiene-1,2, butene-1, butadiene-1,3, cyclopentadiene-1,3 et benzene. Le radical hexadienyle est vraisemblablement un intermediaire de cyclisation. Il est prepare en solution et en phase gazeuse; son comportement chimique est etudie

A variant form of dipeptidyl peptidase II (DPP II), initially reported under the name of "DPP V" (Eisenhauer, D. A., and McDonald, J. K. (1982) Fed. Proc. 41, 507), was detected in aqueous extracts of porcine ovaries on the basis of a markedly enhanced action on prolyl bonds. This porcine form of DPP II, which was most sensitively assayed on Phe-Pro-2-naphthylamide (Phe-Pro-NNap), was purified 1400-fold to a specific activity of 28 mumol/min/mg of protein (pH 6.0, 37 degrees C) from an aqueous extract of hog ovaries taken during pregnancy, when ovarian levels of DPP II are some 3- to 8-fold higher. Purification involved ammonium sulfate fractionation, molecular exclusion chromatography, chromatofocusing, affinity chromatography on concanavalin A-Sepharose 4B, and high performance ion exchange chromatography. The purified enzyme, which was apparently electrophoretically homogeneous at pH 3, 7, and 8.8 (pI = 5.0), was shown to be an Mr = 110,000 glycoprotein containing about 2% carbohydrate (primarily mannose) and less than 1% sialic acid, and to consist of two noncovalently linked Mr = 54,000 subunits. A serine catalytic mechanism was supported by inhibitor studies and by common mobilities seen during electrophoresis for (histochemically detected) Phe-Pro-arylamidase activity and the [14C]diisopropyl fluorophosphate-labeled enzyme. In the standard fluorometric assay at 37 degrees C, Phe-Pro-NNap (0.2 mM) was hydrolyzed optimally at pH 6.0 (Km = 45 microM; kcat = 54 s-1). In comparison to the rate seen on Lys-Ala-NNap, the usual DI P II assay substrate, rates seen on Phe-Pro-, Lys-Pro-, and Arg-Pro-NNap were about 8-, 4-, and 2-fold higher, respectively. No action occurred on N-blocked derivatives or on Pro-NNap. Action on oligopeptides appeared to be limited to tripeptides, in particular those containing proline or alanine in the P1 position, i.e. Phe-Pro-Ala (100%), Lys-Ala-Ala (35%), Ala-Ala-Ala (33%), and Gly-Pro-Ala (26%). Only a trace of activity was seen on Phe-Pro-Ala-Ala, and none on Z-Phe-Pro-Ala or Ala-Ala-Ala-OMe. Evidence for the lysosomal localization of DPP II included sedimentability and latency and its distribution, coincident with acid phosphatase, in two distinct isopycnic regions following equilibrium density centrifugation. Lysosomal heterogeneity was suggested by this dual isopycnic banding.(ABSTRACT TRUNCATED AT 400 WORDS)

The rat kidney cortex was found to contain two N-terminal exopeptidases of the tripeptidyl peptidase (TPP) class. Each required a free N-terminus to catalyze the release of collagen-related (Gly-Pro-X) "triplets." In accordance with their apparent pH optima, activities were routinely determined fluorimetrically at pH 4.0 (TPP 4) and at pH 7.0 (TPP 7) on Gly-Pro-Met-2-naphthylamide. The specific activity in both the homogenate and the classical subfractions was much greater at pH 7 than at pH 4. Subfractionation of the microsomal fraction by equilibrium banding in sucrose did not separate the TPP 4 and TPP 7 activities. The banding density (1.18 g/ml) and the distribution patterns for TPP 7 in the microsomal subfractions, and also in the subfractions of the small lysosomes in the mitochondrial-lysosomal (ML) fraction, demonstrate that TPP 7 is associated with smooth membranes. The TPP 4 and TPP 7 activities were clearly separated during subfractionation of the ML fraction. Rate sedimentation demonstrated that TPP 4 was present in the large, fast-sedimenting lysosomes (protein droplets) and in a heterogeneous broad band of smaller lysosomes. Equilibrium banding of the small lysosomes gave two distinct TPP 4-containing populations at densities 1.20 and 1.235 g/ml. Notably, dipeptidyl peptidase II (DPP II) gave identical banding densities and showed distributions very similar to TPP 4.(ABSTRACT TRUNCATED AT 250 WORDS)

The somatostatin-related peptides somatostatin-14 (SS-14) and somatostatin-28 (aSS-28) are synthesized at the C-terminal end of two separate pre-pro-somatostatins in anglerfish pancreatic islets. The purpose of this study was to determine whether these peptides are expressed in the same or different cell types. Antisera R141 and R293, which recognize the central region of SS-14 and the C-terminal region of aSS-28 ([Tyr7,Gly10] SS-14), respectively, were used in an immunohistochemical examination of anglerfish islets. The R293 antiserum-labeled cells were distributed individually or in small clusters. These same cells, as well as a separate set of cells arranged in large clusters, were stained by the R141 antiserum. Pre-absorption of the R141 antiserum with [Tyr7,Gly10] SS-14 eliminated staining by R141 of only those cells also labeled by R293, whereas pre-absorption of R141 with SS-14 prevented all staining. Pre-absorption of R293 with [Tyr7,Gly10] SS-14 eliminated all staining, whereas pre-absorption with SS-14 had no effect on aSS-28-like immunoreactivity. These results suggest the existence of two separate cell types which express either SS-14 or aSS-28. The cells that contained the somatostatin-related peptides were found to be distinct from those cells that contained insulin, glucagon, or anglerfish peptide Y. However, the cells stained by the R293 antiserum were distributed in close association with glucagon-containing cells. The implications of the existence of separate cell types which express SS-14 or aSS-28 are discussed with regard to processing of the biosynthetic precursors to these peptides.

Guinea pig spermatozoa were found to contain a fully-latent cysteine proteinase that could be unmasked by incubating epididymal sperm for 2 hr at pH 3.5 and 37 degrees C. The proteinase was identified as cathepsin L (EC 3.4.22.15) on the basis of its optimal hydrolysis of benzyloxycarbonyl-Phe-Arg-7-(4-methyl)coumarylamide (Z-Phe-Arg-NMec) at pH 5.5; lack of action on Z-Arg-Arg-NMec and Arg-NMec; urea-enhanced digestion of azocasein; marked sensitivity to thiol reagents, leupeptin, Z-Phe-Phe-CHN2, and L-trans-epoxy-succinylleucylamido(3-methyl)butane (Ep-475 or E-64-c); and insensitivity to pepstatin and serine proteinase inhibitors. Gossypol, a male antifertility agent, was inhibitory. The unmasking phenomenon was reversibly inhibited by HgCl2 and mersalyl acid, and prevented by leupeptin and Ep-475, but not by pepstatin.

Substrate specificity of cathepsin C derived from rat liver, describing polymeric structure and behavior as acidic protein

Purification and substrate specificity of dipeptidyl arylamidase II from extracts of bovine anterior pituitary glands, noting peptidase and dipeptide esterase activities

An enzyme is described that was observed in extracts of rat and bovine pituitary glands. This enzyme, named dipeptidyl arylamidase II, catalyzes the cleavage of the dipeptide moiety from Lys-Ala-β-naphthylamide at pH 5.5, but not from monoamino acid arylamides. Dipeptidyl arylamidase II exhibits no metal, halide, or sulfhydryl dependences, and its specificity is restricted to substrates having an unsubstituted NH2-terminus. Lower but significant rates of hydrolysis were also exhibited at pH 5.5 on the β-naphthylamides of Arg-Ala and Leu-Ala, with only traces of activity on the arylamides of Ala-Ala, Gly-Pro, and Ser-Met. No activity was detected on the β-naphthylamides of benzylodicarbonyl-Lys-Ala, Lys-Lys, Arg-Arg, Gly-Arg, Gly-Phe, Ser-Tyr, or His-Ser. Cations were exceptionally inhibitory to the hydrolysis of Lys-Ala-β-naphthylamide by dipeptidyl arylamidase II, and the sensitivity of the enzyme increased with the size of the cation: Li+ l Na+ l K+ l Tris l puromycin aminonucleoside l puromycin. A 50 mm concentration of Tris, pH 5.5, in the assay system gave a 96% inhibition of arylamidase II activity, while puromycin at 1 mm gave a 76% inhibition. A Km of 1.1 x 10-5 m was determined for the hydrolysis of Lys-Ala-β-naphthylamide at pH 5.5 at 37°. The inhibition by cations was competitive for all cations tested, with Ki values at pH 5.5 and 37° of 1.8 x 10-3 m for Na+, 3.2 x 10-4 m for Tris, 1.9 x 10-5 m for puromycin, and 3.0 x 10-5 m for puromycin aminonucleoside. Dipeptidyl arylamidase II activity was detected in tissues other than pituitary; thyroid, spleen, and kidney were relatively rich sources, whereas liver, pancreas, and serum were poor. The hydrolysis of Lys-Ala-β-naphthylamide at pH 5.5 by all tissues tested showed the same sensitivity to Tris ions. As indicated by studies with sucrose homogenates of fresh rat pituitary glands, and with subcellular fractions prepared by differential and density gradient centrifugation, dipeptidyl arylamidase II was located, along with acid phosphatase, in pituitary lysosomes. However, unlike acid phosphatase, arylamidase II appeared to be located almost exclusively in the lysosomes, and exhibited a functional latency of about 91 to 95%. Ser-Tyr arylamidase (dipeptidyl arylamidase I) was also found to be lysosomal, whereas Arg-Arg arylamidase (dipeptidyl arylamidase III) was cytoplasmic. The aminopolypeptidase and dipeptidase activities measured on Lys- and Arg-β-naphthylamide were predominantly (90 to 95%) cytoplasmic.

Azulene is an aromatic hydrocarbon that, in addition to being an isomer of naphthalene, is also isoelectronic with it.

A variant form of dipeptidyl peptidase II (DPP II), initially reported under the name of "DPP V" (Eisenhauer, D. A., and McDonald, J. K. (1982) Fed. Proc. 41, 507), was detected in aqueous extracts of porcine ovaries on the basis of a markedly enhanced action on prolyl bonds. This porcine form of DPP II, which was most sensitively assayed on Phe-Pro-2-naphthylamide (Phe-Pro-NNap), was purified 1400-fold to a specific activity of 28 mumol/min/mg of protein (pH 6.0, 37 degrees C) from an aqueous extract of hog ovaries taken during pregnancy, when ovarian levels of DPP II are some 3- to 8-fold higher. Purification involved ammonium sulfate fractionation, molecular exclusion chromatography, chromatofocusing, affinity chromatography on concanavalin A-Sepharose 4B, and high performance ion exchange chromatography. The purified enzyme, which was apparently electrophoretically homogeneous at pH 3, 7, and 8.8 (pI = 5.0), was shown to be an Mr = 110,000 glycoprotein containing about 2% carbohydrate (primarily mannose) and less than 1% sialic acid, and to consist of two noncovalently linked Mr = 54,000 subunits. A serine catalytic mechanism was supported by inhibitor studies and by common mobilities seen during electrophoresis for (histochemically detected) Phe-Pro-arylamidase activity and the [14C]diisopropyl fluorophosphate-labeled enzyme. In the standard fluorometric assay at 37 degrees C, Phe-Pro-NNap (0.2 mM) was hydrolyzed optimally at pH 6.0 (Km = 45 microM; kcat = 54 s-1). In comparison to the rate seen on Lys-Ala-NNap, the usual DI P II assay substrate, rates seen on Phe-Pro-, Lys-Pro-, and Arg-Pro-NNap were about 8-, 4-, and 2-fold higher, respectively. No action occurred on N-blocked derivatives or on Pro-NNap. Action on oligopeptides appeared to be limited to tripeptides, in particular those containing proline or alanine in the P1 position, i.e. Phe-Pro-Ala (100%), Lys-Ala-Ala (35%), Ala-Ala-Ala (33%), and Gly-Pro-Ala (26%). Only a trace of activity was seen on Phe-Pro-Ala-Ala, and none on Z-Phe-Pro-Ala or Ala-Ala-Ala-OMe. Evidence for the lysosomal localization of DPP II included sedimentability and latency and its distribution, coincident with acid phosphatase, in two distinct isopycnic regions following equilibrium density centrifugation. Lysosomal heterogeneity was suggested by this dual isopycnic banding.(ABSTRACT TRUNCATED AT 400 WORDS)

Relaxin, a peptide hormone responsible for the widening of the birth canal in mammals, has been purified from the ovaries of pregnant hogs. The amino acid sequences of its constituent A and B chains were determined, and the positions of the disulfide cross-links were established. Relaxin was shown to be identical to insulin with respect to its disulfide bond distribution, but significant homology was lacking in other positions. These findings suggest that relaxin and insulin were derived from a common ancestral gene. Since the intrauterine mode of propagation is synonymous with the development of mammals, the genetic distance between insulin and relaxin should therefore permit an estimate of the earliest possible time of commitment of one evolutionary branch to the development of mammals. This event was estimated to have occurred about 5 X 10(8) years ago.

Dipeptidyl aminopeptidase II (DAP II) was demonstrated cytochemically at light and electron microscope levels in rat macrophages and mast cells using Lys-Ala-4-methoxy-2-naphthylamide as a specific substrate. The enzyme which was found to be lysosomal in both cell types, was analyzed biochemically in extracts by measuring fluorometrically the liberated naphthylamine, and was visualized in sections microscopically using azo-coupling methods. DAP II was further characterized by isoelectric focusing techniques. Macrophage DAP II was found to be typical of that found in other rat tissues in terms of its structural latency, substrate specificity, inhibitor sensitivities, and pH activator requirements. Addition DAP II isozymes, not previously recognized, were observed.

L'irradiation (40 torr et 100 u/cm 2 ) de l'hexadiene-1,5 conduit aux methane, ethylene, acetylene, propene, propadiene-1,2, butene-1, butadiene-1,3, cyclopentadiene-1,3 et benzene. Le radical hexadienyle est vraisemblablement un intermediaire de cyclisation. Il est prepare en solution et en phase gazeuse; son comportement chimique est etudie

Calcitonin gene-related peptide (CGRP) has been found in nerves that innervate the rat ovary. In this study we used indirect immunohistochemical methods to investigate the normal distribution of CGRP-immunoreactive fibers in the prepubertal rat ovary and to determine the route by which these fibers reach the gland. Additional experiments were performed to examine the possible colocalization of CGRP with immunoreactivities of substance P-, neuropeptide Y-, and tyrosine hydroxylase. Potential effects of CGRP on estradiol and progesterone secretion were explored with cultured granulosa cells and short-term incubation of whole ovaries in vitro. In ovaries with intact nerves, CGRP-labeled fibers were observed in dense plexuses surrounding the ovarian vasculature. Additional fibers were found occasionally in the interstitial tissue and in the vicinity of ovarian follicles. Surgical transection of the plexus nerve completely eliminated CGRP immunoreactivity. Section of the superior ovarian nerve or the abdominal vagal trunks had no discernible effect on CGRP labeling. Additional studies revealed coexistence of CGRP and substance P in several axons. Neither tyrosine hydroxylase nor neuropeptide Y-immunoreactivity was co-localized in CGRP-containing fibers. CGRP (10(-10) - 10(-6) M) had no effect on the basal or follicle-stimulating hormone-stimulated release of estradiol or progesterone from ovaries of pregnant mare's serum gonadotropin-treated rats or from cultured granulosa cells from hypophysectomized, diethylstilbestrol-treated rats. We conclude that CGRP-containing nerves enter the ovary via the plexus nerve and are probably involved in the regulation of vasomotor function.

NPY exhibits a broad distribution throughout the body. NPY has been localized in neurons that synthesize norepinephrine or epinephrine and also in many cell bodies which are not catecholaminergic. Coexistence of NPY and several other peptides has also been observed. Accordingly, NPY displays a wide variety of functional activities depending on its location and coexistence with other substances, especially catecholamines. NPY exerts direct effects at several targets and also modulates the cellular response to catecholamines and to peptides such as LHRH. It is reasonable to expect that NPY will modulate the pre- and postjunctional effects of catecholamines at many of their targets in view of the distribution of NPY in central catecholaminergic neurons and throughout the preaortic and sympathetic chain ganglia. Virtually nothing is known at the time of this writing about NPY receptors and postreceptor transduction mechanisms at different sites of NPY activity. Equally mysterious are the pre- and postjunctional receptor-coupled transduction mechanisms which are involved in the modulation of catecholaminergic or other peptidergic effects by NPY. The distribution of NPY and its involvement in cardiovascular, GI, endocrine, and neuroendocrine systems suggest that NPY may be an extremely important regulator of a spectrum of physiological functions.

Relaxin, a peptide hormone responsible for the widening of the birth canal in mammals, has been purified from the ovaries of pregnant hogs. The amino acid sequences of its constituent A and B chains were determined, and the positions of the disulfide cross-links were established. Relaxin was shown to be identical to insulin with respect to its disulfide bond distribution, but significant homology was lacking in other positions. These findings suggest that relaxin and insulin were derived from a common ancestral gene. Since the intrauterine mode of propagation is synonymous with the development of mammals, the genetic distance between insulin and relaxin should therefore permit an estimate of the earliest possible time of commitment of one evolutionary branch to the development of mammals. This event was estimated to have occurred about 5 X 10(8) years ago.

The amino acid sequence of cystatin, the protein from chicken egg-white that is a tight-binding inhibitor of many cysteine proteinases, is reported. Cystatin is composed of 116 amino acid residues, and the Mr is calculated to be 13 143. No striking similarity to any other known sequence has been detected. The results of computer analysis of the sequence and c.d. spectrometry indicate that the secondary structure includes relatively little alpha-helix (about 20%) and that the remainder is mainly beta-structure.