Self-rated sleep characteristics and bother from nocturia.

PURPOSE: The aim of this study was to evaluate if men with varying degrees of bother from a similar number of nocturia episodes differ with respect to self-rated sleep characteristics and fatigue. MATERIALS AND METHODS: As part of the baseline assessments during a nocturia treatment trial, 55 participants reported frequency and bother of nocturia using the AUA Symptom Inventory and completed 7-day sleep diaries prior to treatment. Participants who reported moderate nocturia (either two or three episodes nightly) were further grouped into categories of LOW (nocturia is no problem or a very small problem) or HIGH bother (nocturia is a big problem). Information from the participant completed sleep diaries was abstracted, including information on daytime napping, total sleep time, mean time needed to return to sleep, nighttime ratings of fatigue, and daytime ratings of fatigue. RESULTS: Of the 55 individuals who completed the pilot study, 24 study participants reported two or three episodes of nocturia and had either HIGH (n = 11) or LOW (n = 13) bother. Participants categorised with HIGH bother were significantly more likely than those with LOW bother to report difficulty initiating sleep (47.7 ± 34.4 vs. 23.5 ± 13.6 min, p = 0.05), difficulty returning to sleep after an awakening (28.9 ± 16.1 vs. 15.4 ± 9.6 min, p = 0.03) and greater morning fatigue (3.3 ± 0.7 vs. 2.5 ± 1.0, p = 0.04 on a 7-point scale). CONCLUSIONS: Since bother related to nocturia is linked to sleep quality, interventions targeting fatigue and sleep maintenance may provide useful targets in the management of nocturia in men.

LYSOZYME IN EPIPHYSEAL CARTILAGE : IV. Embryonic Chick Cartilage Lysozyme-Its Localization and Partial Characterization.

THE LOCALIZATION OF CHICK EMBRYONIC LYSOZYME WAS DETERMINED BY TWO TECHNIQUES: by studying the rate of release from the tissue during sequential enzymatic digestion and by immunocytochemistry. Both techniques indicate that, in this tissue, lysozyme is primarily extra-cellular. Cartilage lysozyme was isolated and partially characterized and found to be identical with egg white lysozyme in its immunologic and enzymatic behavior. In addition, a method for the isolation of large numbers of viable chondrocytes is described.

ENZYME ACTIVITIES IN CHICK EMBRYONIC CARTILAGE : Their Subcellular Distribution in Isolated Chondrocytes.

Some characteristic enzymatic activities were determined in chick embryonic cartilage and compared with the analogous activities in bone and liver. Chondrocytes were isolated, broken by sonication, and subjected to subcellular fractionation to yield a nuclear pellet, the mitochondrial, lysosomal, and microsomal fractions, and the high speed supernatant solution. It was established that these fractions are characterized by enzymatic activities usually associated with similar fractions in other tissues, but with some quantitative differences. Lysozyme, a particulate-associated enzyme in other tissues, was not detected in any subcellular fraction even by the sensitive technique of microzone electrophoresis and is therefore considered to be primarily extracellular in cartilage.

Lysozyme in epiphyseal cartilage. II. The effect of egg white lysozyme on mouse embryonic femurs in organ cultures.

Embryonic mouse femoral cartilage, like the epiphyseal cartilage of the calf scapula, contains large amounts of lysozyme. The addition of egg white lysozyme to organ cultures of embryonic mouse femurs induces unique alterations in the gross and microscopic morphology of the femurs. The sites of these alterations are precisely related to the natural distribution of lysozyme in calf scapula. If the exogenous lysozyme is withdrawn from the culture, the morphological changes disappear, accompanied by a resumption or derepression of growth. The effect on growth is evident only in 17-day embryos. These observations support the idea that lysozyme has a physiological role in cartilage, perhaps related to a regulatory mechanism in bone formation.

Both cell substratum regulation and hormonal regulation of milk protein gene expression are exerted primarily at the posttranscriptional level.

The mechanism by which individual peptide and steroid hormones and cell-substratum interactions regulate milk protein gene expression has been studied in the COMMA-D mammary epithelial cell line. In the presence of insulin, hydrocortisone, and prolactin, growth of COMMA-D cells on floating collagen gels in comparison with that on a plastic substratum resulted in a 2.5- to 3-fold increase in the relative rate of beta-casein gene transcription but a 37-fold increase in beta-casein mRNA accumulation. In contrast, whey acidic protein gene transcription was constitutive in COMMA-D cells grown on either substratum, but its mRNA was unstable and little intact mature mRNA was detected. Culturing COMMA-D cells on collagen also promoted increased expression of other genes expressed in differentiated mammary epithelial cells, including those encoding alpha- and gamma-casein, transferrin, malic enzyme, and phosphoenolpyruvate carboxykinase but decreased the expression of actin and histone genes. Using COMMA-D cells, we defined further the role of individual hormones in influencing beta-casein gene transcription. With insulin alone, a basal level of beta-casein gene transcription was detected in COMMA-D cells grown on floating collagen gels. Addition of prolactin but not hydrocortisone resulted in a 2.5- to 3.0-fold increase in beta-casein gene transcription, but both hormones were required to elicit the maximal 73-fold induction in mRNA accumulation. This posttranscriptional effect of hormones on casein mRNA accumulation preceded any detectable changes in the relative rate of transcription. Thus, regulation by both hormones and cell substratum of casein gene expression is exerted primarily at the post transcriptional level.

Effect of medroxyprogesterone and an aorta-derived cell growth inhibitor on B16 melanoma in mice.

An aorta-derived inhibitor of endothelial cell and tumor cell growth and medroxyprogesterone, which depresses collagenase expression in vivo, were tested alone and in combination against B16-F10 melanoma in C57BL/6 mice in such doses that either agent alone had little effect. Together, these agents retarded growth of subcutaneously transplanted tumor cells and reduced the number and size of pulmonary tumors after iv tumor cell injection. Of the treatments used, only the aortic factor administered alone prolonged life in mice with pulmonary tumors.

Heparin-binding fragments of fibronectin are potent inhibitors of endothelial cell growth: structure-function correlations.

Heparin-binding fragments derived from the amino- and carboxyl-terminal regions of human plasma fibronectin appear to be at least relatively specific potent inhibitors of the growth of bovine aortic endothelial cells in culture by as yet unknown mechanisms. In order to understand better the sites which subserve this activity, we have compared the relative potency of other major fragments of fibronectin, most of which have dissimilar properties and do not bind heparin. We have also proteolytically digested and chemically modified the most potent of these fragments, the amino-terminal 29-kDa fragment, in order to test whether structural alterations that affect heparin-binding also affect the inhibitory property. Not all chemical modifications that abolished heparin-binding also abolished endothelial cell growth. Neither an amino-terminal 20-kDa nor a carboxyl-terminal 8-kDa subfragment of the 29-kDa fragment bound heparin; however, both were as inhibitory as native 29-kDa fragment. Reduction of the disulfides of the 20-kDa and 8-kDa fragments did not abolish inhibitory activity. We therefore conclude that the activity is not strictly conformation-dependent and that although the inhibitory activity is distributed throughout the 29-kDa segment, it can be expressed by an 8-kDa carboxyl-terminal segment containing residues of the last Type I loop structure.

Angiogenesis in arteries: review.

The inner parts of the walls of large blood vessels do not normally contain intrinsic vasculature. In pathologic conditions such as arteriosclerosis or thrombosis, angiogenesis occurs, and may have significant clinical consequences. This review attempts to relate the little that is known about the factors specific to vascular walls which regulate angiogenesis to more general knowledge of the phenomenon.

The ground substance of the arterial wall. Part 2. Electron-microscopic studies.

Electron microscopy of ruthenium red stained bovine aorta before and after chondroitinase digestion demonstrates proteoglycans on and between collagen fibrils. The collagen-associated proteoglycans include a proteoglycan previously purified from this tissue as demonstrated by immunocytochemistry and are extractable with high molar guanidine HC1. In loci rich in proteoglycans such as areas of turbulent flow in calves, more proteoglycan can be demonstrated morphologically, and these molecules also coat elastin.

The effect of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) on a rabbit model of athero-arteriosclerosis.

The purpose of this study was to test the effectiveness of various doses of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) in an experimental rabbit model of athero-arteriosclerosis designed by Hass et al. (Amer. J. Pathol., 49 (1966) 739). This model, which involves the feeding of a hypercholesterolemic diet in conjunction with the administration of moderately high doses of vitamin D and nicotine, results in an extensive arterial disease with complicated lesions. EHDP was administered daily by subcutaneous injection at levels of 0.25, 1.0 and 2.5 mg/kg body weight beginning with the initiation of the atherogenic regimen. Results of chemical and histopathological analyses after 8 and 12 weeks of treatment indicate the following: (1) There was a dose-related inhibition of arterial calcification at 8 weeks. At 12 weeks, only the 2.5 mg/kg dose of EHDP resulted in reduced calcification. (2) EHDP administration appeared to influence arterial lipid-containing plaque formation in medium sized arteries at 12 weeks. There was no apparent effect of EHDP administration on serum cholesterol and triglyceride levels. (3) EHDP, at a dose of 2.5 mg/kg/day, inhibited the vitamin D induced hypercalcemia. (4) EHDP administration at 2.5 mg/kg/day almost totally inhibited the thromboarteritis accompanying this disease. (5) The data thus indicate that if arterial calcification is inhibited, the other morphological effects of this treatment regime are also inhibited. This effect occurred even though serum lipid levels were unaffected. The data therefore emphasize the role of calcification in the pathogenesis of this type of experimental atherosclerosis and perhaps in human disease as well.

The ground substance of the arterial wall. Part 1. Extractability of glycosaminoglycans and the isolation of a proteoglycan from bovine aorta.

Water and glycosaminoglycan contents were measured in upper and lower thoracic aortas of claves and steers. The ability of various molarities of guanidine hydrochloride to extract glycosaminoglycans from these tissues was assessed. Some glycosaminoglycans seem to be more resistant to extraction than others. A procedure is described for the isolation of a proteoglycan. The molecule appears to contain both dermatan sulfate and chondroitin sulfate. It also seems to be less dense than cartilage proteoglycans extracted by similar methods as assessed by its behavior in centrifugal fields. The properties, locus and biological activities of this molecule are currently being studied.

Immunologic studies of bovine aortic and cartilage proteoglycans.

Rabbit antisera prepared against bovine cartilage and aortic proteoglycan monomer (PGM) were employed to identify and localize the distribution and cross-species reactivities of these PGMs. A specific reaction to both cartilage and aortic PGM was obtained as revealed by rocket immunoelectrophoresis of the immunogens against the respective antisera, with no reaction against bovine serum albumin or human fibronectin. These antisera appear to demonstrate tissue specificity. Indirect immunofluorescence studies with antiserum to aortic PGM on bovine aorta tissue revealed intense localization of fluorescence in the intima and superficial media with staining alone collagen fibers, around smooth muscle cells, and on the surface of elastin. When antiserum to cartilage PGM was applied to bovine aorta, staining was much weaker and localized more in the interfibrillar matrix. On the other hand, when antiserum to aortic PGM was used to stain bovine nasal septum cartilage, fluorescent staining was restricted to the pericellular matrix, in contrast to the diffuse, intense staining of both pericellular and extraterritorial staining by antiserum to cartilage PGM. Absorption with eigher bovine aorta or cartilage PGM abolished antibody activity against the absorbing molecules only. The immunologic distinction between the pericellular and extraterritorial cartilage matrix was demonstrated further when both antisera were studied with guanidine chloride (GuHCl)-extracted bovine nasal septum cartilage, when they both demonstrated the same pericellular matrix staining. Despite the tissue specificity, these antisera are not completely species specific, since staining in a similar pattern was observed in both human and rat tissues.

Discovery of the ceruloplasmin homologue hephaestin: new insight into the copper/iron connection.

Approximately 75 years ago Hart and colleagues discovered that copper deficiency impaired mammalian iron metabolism. Discovery of hephaestin identifies a critical new component of the copper and iron connection in mammals. Hephaestin appears to be a multicopper oxidase required for efficient export of iron from the intestine.

Interaction of the hemochromatosis gene product HFE with transferrin receptor modulates cellular iron metabolism.

Mutations of a novel MHC class I-like protein, termed HFE, have been found in the vast majority of patients with the iron overload disease heredity hemochromatosis. Identification of HFE is likely to shed light on one of the major enigmas of mammalian iron homeostasis: How is intestinal iron absorption regulated?

The inhibition of corneal vascularization in rabbits.

A single subconjunctival injection of 250 microgram of a partially purified extract of bovine aorta, administered immediately before or after silver nitrate injury to the cornea, markedly inhibited corneal vascularization in the rabbit eye. We believe the active molecule is a protease inhibitor that prevents the potential source of new vessels from proliferating and invading the diseased cornea.

The inhibition of corneal vascularization with aortic extracts in rabbits.

A low molecular weight fraction of bovine aortic extract inhibited corneal vascularization and edema in rabbits when administered either subconjunctivally or topically as long as 48 hours after injury. The extract also appeared to enhance the regression of newly formed corneal vessels. Topical administration for as long as two months had no deleterious ocular side effects. Tissue culture experiments showed that analagous fractions prepared from bovine vitreous inhibit endothelial cell growth. The major growth inhibitor of corneal neovascularization was not the Kunitz bovine protease inhibitor.

Inhibitor of vascular endothelial cell growth in the lens.

Because diabetic eyes with proliferative retinopathy show an increased incidence of iris neovascularization after removal of the lens, we studied the effect of extracts of human and bovine lenses on one of the key events in the neovascular process, endothelial cell proliferation. Both human and bovine lens extracts demonstrated a dose-dependent inhibition of bovine aorta endothelial cell proliferation that was greatest in extracts with molecular weight of less than 100,000. This inhibition was reversible and the extracts did not significantly inhibit smooth muscle cell proliferation. Considerable endothelial cell inhibitory activity was present in bovine lens capsule extracts. These data suggested that the lens may play an active role in the prevention of iris neovascularization.

Growth inhibitory activities in avascular tissues are recognized by anti-transforming growth factor beta antibodies.

The aqueous and vitreous humors are normally avascular and may be sites of altered inflammatory responses. Inhibitors of endothelial cell and lymphocyte growth as well as of angiogenesis have been isolated from vitreous and an inhibitor of lymphocyte mitogenesis has been found in aqueous. Data are presented here indicating that aqueous humor also inhibits endothelial cell growth. In addition, neutralizing antibodies against TGF beta block the effects of materials isolated from these tissues on endothelial cell and lymphocyte growth. Similar data were obtained from aorta, another normally avascular site. These observations suggest that the avascularity and altered inflammatory responses in these sites may be regulated in part by TGF beta.