Transdermal estrogen for female stress urinary incontinence in postmenopause.

OBJECTIVE: To evaluate the effect of transdermal estrogen for stress urinary incontinence in postmenopause. STUDY DESIGN: An open within patient, dose-finding study with transdermal 17-beta-estradiol combined with cyclic medroxyprogesterone acetate was conducted over 9 months in 21 patients (mean age 57.3 years) suffering from urodynamically verified mild to moderate stress incontinence without detrusor instability. RESULTS: Subjective improvement was noted in 16 out of 21 patients (76%). The dose level of 50 micrograms was better tolerated than 100 micrograms and sufficient enough to achieve continence. CONCLUSION: Transdermal estrogen therapy plays an adjuvant role in conservative therapy for mild to moderate stress urinary incontinence in postmenopausal women.

Acid cysteine proteinase inhibitor in cutaneous lymphocytic infiltrates.

Acid cysteine proteinase inhibitor (ACPI, cystatin A) is normally present in squamous epithelium and dendritic cells of lymphoid follicles. Its expression is altered both in proliferative and malignant squamous epithelium and in neoplastic lymphoid follicles. The expression of ACPI in the lymphoid infiltrates of cutaneous psuedolymphomas and B-cell lymphomas was studied. Eighteen pseudolymphomas from 15 patients were divided into three groups according to the proportion of B and T lymphocytes. The B-cell-type lesions with well-developed follicles and germinal centers showed a pronounced ACPI expression in dendritic cells. Varying amounts of ACPI-positive cells were present in the mixed B- and T-cell-type and also in the T-cell-type lesions. The labeled cell population was distinct from the factor XIIIa-positive dermal dendrocytes, S-100-positive histiocytes, and HAM 56-positive histiocytes. Malignant lymphomas contained a few haphazardly arranged ACPI-positive cells with short dendrites and granular cytoplasm. It was concluded that follicular dendritic cells can be reliably labeled with ACPI antiserum in cutaneous pseudolymphomas. The structure and distribution of ACPI-containing cells in malignant cutaneous B-cell lymphomas is altered when compared with pseudolymphomas.

Identification of acid cysteine proteinase inhibitor (cystatin A) in the human thymus.

BACKGROUND: Acid cysteine proteinase inhibitor (ACPI, also called cystatin A) is a protein that is present in the epithelial cells of the skin and in the dendritic reticulum cells of lymphoid tissues. In this study the presence and cellular localization of ACPI in the thymus was investigated. METHODS: The cellular and topographical location of ACPI was immunohistochemically demonstrated in the normal thymus of man. RESULTS: ACPI was found in the cells of the Hassall's corpuscles and in many medullary cells. Most of these cells were epithelial cells, as shown by the results of immunohistochemical cytokeratin and epithelial membrane antigen stainings. Also, some individual cytokeratin negative but S-100 positive medullary reticular dendritic cells were stained with ACPI. CONCLUSIONS: The finding that ACPI is constantly present in the thymus at restricted and specific cellular locations leads to the suggestion that protease inhibitors may play a role in specific thymic functions.

Cystatin A-like immunoreactivity is widely distributed in human brain and accumulates in neuritic plaques of Alzheimer disease subjects.

The cellular localization of cystatin A, an endogenously occurring inhibitor of lysosomal thiol proteases (cathepsins B, H, L and S), was studied immunohistochemically in human postmortem brain using the peroxidase-antiperoxidase method. Both polyclonal and monoclonal antibodies to cystatin A were employed. Western blot analysis revealed one molecular form of the inhibitor in human brain extracts. Its molecular weight was about 13,000. Immunostaining appeared in a sizeable population of neurons and a few cells surrounding cerebral blood vessels (pericytes). In Alzheimer disease subjects cystatin A was found in many neuritic plaques. Possible functional consequences with regard to a role of cystatin A in the inhibition of the Alzheimer amyloid precursor protein (APP)-clipping enzyme, cathepsin B, are discussed.

Tight-binding inhibition of cathepsin S by cystatins.

Human cystatins A, B and C were purified, and their inhibition efficiency was tested with the cysteine proteinase cathepsin S. Cathepsin S was strongly inhibited by cystatins A and B in the subnanomolar range and by cystatin C in the picomolar range. Two steps of inhibition of cathepsin S by the cystatins which involve slow binding are discussed.

Protein Synthesis during Natural and Precocious Soybean Seed (Glycine max [L.] Merr.) Maturation.

Protein synthesis was studied during precocious and natural soybean seed (Glycine max [L.] Merr.) maturation. Developing seeds harvested 35 days after flowering were precociously matured through controlled dehydration. Total soluble proteins and proteins labeled with [(35)S]methionine were extracted from control, developing seeds and from precociously and naturally matured seeds and were analyzed by one-dimensional PAGE and fluorography. The results demonstrated that several polypeptides which were designated "mature polypeptides," were synthesized de novo during precocious and natural seed maturation. Two of these polypeptides, 31 and 128 kilodalton in mass, also stained intensely with Coomassie blue, suggesting their abundant accumulation during seed maturation. Results from in vitro translation experiments showed that the mRNAs corresponding to these "maturation polypeptides" accumulated during precocious maturation and in naturally matured seeds, but not in seeds freshly harvested 35 days after flowering (control). The role of the "maturation polypeptides" is currently unknown; however, their presence and that of their corresponding mRNAs was coincident with the ability of matured seeds to establish seedling growth. This study has demonstrated that precocious seed maturation treatments may be extremely useful for investigations of metabolic events and molecular control mechanisms affecting soybean seed maturation.

Cystatin C containing neurons in human postmortem hypothalamus.

The regional distribution and cellular localization of cystatin C in neurons of human postmortem hypothalamic was studied by use of peroxidase-anti-peroxidase technique. Cystatin C (earlier named gamma-trace) was found to be present in multiple nerve cells belonging to nuclei supraopticus, paraventricularis and arcuatus. We speculate that the occurrence of cystatin C in human cerebrospinal fluid is the result of a release of the protein from these neurons into the ventricular system.

Changes in Soybean (Glycine max [L.] Merr.) Glycerolipids in Response to Water Stress.

Soybean (Glycine max [L.] Merr.) plants with the first trifoliate leaf fully expanded were exposed to 4 and 8 days of water stress. Leaf water potentials dropped from -0.6 megapascal to -1.7 megapascals after 4 days of stress; then to -3.1 megapascals after 8 days without water. All of the plants recovered when rewatered. The effects of short-term drought stress on triacylglycerol, diacylglycerol, phospholipid, and galactolipid metabolism in the first trifoliate leaves was determined. Leaf triacylglycerol and diacylglycerol content increased 2-fold during the first 4 days of stress and returned to control levels 3 days after rewatering. The polar lipid fraction, which contained phospholipids and galactolipids, changed little during this time. The linolenic acid (18:3) content of the triacylglycerol and diacylglycerol increased 25% during stress and the polar lipid 18:3 content decreased 15%. The pattern of glycerolipid labeling, after applying [2-(14)C]acetate to intact leaves was altered by water stress. After 4 days of water stress the radioactivity of phosphatidic acid + phosphatidylinositol, phosphatidylcholine, triacylglycerol, and diacylglycerol increased between 4 and 9% (compared to control plans) while radioactivity of phosphatidylethanolamine, monogalactosyldiglyceride, and digalactosyldiglyceride decreased 2 to 11%. These data indicated that increased levels of triacylglycerol and diacylglycerol observed during water stress were attributed to de novo synthesis rather than breakdown or reutilization of existing glycerolipids and fatty acids.

A Comparison of Oleic Acid Metabolism in the Soybean (Glycine max [L.] Merr.) Genotypes Williams and A5, a Mutant with Decreased Linoleic Acid in the Seed.

The metabolism of oleoyl coenzyme A (CoA) was examined in developing seed from two soybean (Glycine max [L.] Merr.) genotypes: Williams, a standard cultivar and A5, a mutant containing nearly twice the oleic acid (18:1) content of Williams. The in vitro rates of esterification of oleoyl-CoA to lysophosphatides by acyl-CoA: lysophosphatidylcholine acyltransferase was similar in both genotypes and lysophosphatidyl-ethanolamine was a poor substrate. Crude extracts desaturated exogenous [1-(14)C]dioleoyl phosphatidylcholine at 14% of the rate achieved with [1-(14)C]oleoyl-CoA, and 50 micromolar lysophosphatidylcholine. The desaturase enzyme also required NADH for full activity. Extracts from Williams contained 1.5-fold more oleoyl phosphatidylcholine desaturase activity, on a fresh weight basis, than did A5 and appeared to have a similar affinity for oleoyl-CoA. There was 1.2- to 1.9-fold more linoleic acid (18:2) in phosphatidylcholine from Williams than from A5, measured at two stages of development, but both genotypes had a similar distribution of fatty acids in the one and two positions. Phosphatidylethanolamine in A5 contained relatively more linoleic acid (18:2) in the one position than did Williams. The increased oleic acid (18:1) content in A5 appeared to be a result of decreased rates of 18:1 desaturation of oleoyl-phosphatidylcholine in this genotype.

Life support for 10 weeks with successful fetal outcome after fatal maternal brain damage.

A 31 year old woman in whom subarachnoid and intracerebral haemorrhage occurred during the second trimester of pregnancy was sustained in intensive care with a respirator for 10 weeks. Computed tomography of the brain showed bilateral intraventricular haemorrhages. Because of drug resistant hypotonic episodes at 31 weeks' gestation caesarean section was performed, and a boy was delivered. The woman died of spontaneous cardiac arrest two days after caesarean section, and the boy showed normal development. Life support can be continued for several weeks in a modern intensive care unit after fatal insult to the brain even in a pregnant woman without affecting the fetus.

Studies on the metabolism of lipid molecular species in immature soybean cotyledons.

Metabolism of lipid molecular species in soybean cotyledons (Glycine max [L.] Merr. var. "Harosoy 63") was determined from incorporation studies with radioactive acetate and glycerol. Lipid synthetic activity was highest in immature cotyledons at 30 days after flowering. Distinct differences in labeling patterns of molecular species within lipid classes demonstrated that selective utilization of diglyceride intermediates occurred in complex lipid biosynthesis in soybean. The phospholipid molecular species in this tissue that displayed the highest turnover rates had the following acyl combinations: saturate-linoleic and dioleic in phosphatidic acid; saturate-oleic in phosphatidylinositol and phosphatidylethanolamine; dioleic in phosphatidylcholine; oleic-dilinoleic in N-acylphosphatidylethanolamine. Saturate-dilinoleic, oleic-dilinoleic, trioleic, and trilinoleic structures were rapidly synthesized species of triglyceride in immature soybean cotyledons.

Lipid molecular species composition in developing soybean cotyledons.

The fatty acid composition of triglyceride and phospholipids in developing soybean cotyledons (Glycine max L., var. "Harosoy 63") was analyzed at several stages of growth between 30 and 70 days after flowering. Changes observed in fatty acid composition within each lipid class were related to the levels of lipid molecular species present in the oil. Thirteen molecular species of triglyceride were identified in developing cotyledons, however three of these groups: trilinolenic, dilinolenic-monolinoleic, and linolenic-linoleic-oleic triglycerides, were not found in the mature seed. In immature cotyledons, trioleic and trilinoleic triglycerides accounted for 50% of the structures found; the level of these molecules decreased to 24.9% in the mature seed. The dilinoleic-monolinolenic triglycerides increased from 0.4 to 23.4% during cotyledon development. Changes in triglyceride composition were compared to the levels of molecular species for each phospholipid class. Dilinoleic and monosaturated monolinoleic phospholipid species were dominant in all phospholipid classes throughout development.

Involvement of phospholipids in triglyceride biosynthesis by developing soybean cotyledons.

The incorporation of phospholipids specifically labeled with glycerol-2(3)H and acyl-(14)C by whole cell tissues of developing soybean cotyledons (Glycine max L.) reveals that phosphatidylinositol, phosphatidylcholine, phosphatidylethanolamine, N-acylphosphatidylethanolamine, and phosphatidic acid can be metabolized to diglyceride. The diglyceride formed may be recylced into phospholipid or acylated to triglyceride. Diglyceride from phosphatidic acid and phosphatidylethanolamine is used readily in triglyceride biosynthesis compared to the other phospholipids. Incorporation of N-acylphosphatidylethanolamine having [9-10-(3)H(N)]oleic acid esterified at sn-3 in cotyledons shows rapid acyltransfer of (3)H into triglyceride and therefore N-acylphosphatidylethanolamine appears to participate in triglyceride biosynthesis as an acyl donor. These studies emphasize phospholipid metabolism in developing soybean cotyledons is a dynamic process which plays a key role in triglyceride formation.

Studies on lipid synthesis and degradation in developing soybean cotyledons.

The metabolic activity of individual lipid classes found in developing soybean cotyledons (Glycine max.) is estimated by determining the degradation rate of the compound under given conditions. Pulse-labeling and dual substrate labeling are used to evaluate this parameter. These studies indicate first order decay kinetics for phosphatidic acid, phosphatidylinositol, phosphatidylcholine, phosphatidylethanolamine, N-acyl-phosphatidylethanolamine, diglyceride, and zero order kinetics for triglyceride in cotyledons var. "Harosoy 63" at 30 days after flowering. Decay coefficients for acyl groups and lipid-glycerol moieties within specific lipid classes from either method are comparable. Half-life (t((1/2))) calculations from the decay coefficients indicate extremely rapid turn-over rates (0.08 to 3.4 hours at 25 C) and suggest similar turnover rates of acyl groups and lipid-glycerol in diglyceride and all phospholipids except N-acylphosphatidylethanolamine where acyl groups are replaced independent of the glycerol moiety. These experiments reveal not only different metabolic activity between lipid components of soybean cotyledons, but also describe a new method for measuring lipid turnover in plants.

Effect of freezing and cold storage on phospholipids in developing soybean cotyledons.

Freezing of plant tissue adversely affects lipid composition. Immature soybean cotyledons (Glycine max L. Merr.) var. "Harosoy 63" were frozen with liquid N(2), dry ice, or stored in a freezer (-20 C) before lipid extraction. The effects of freezing temperature, thawing rate, and cold storage on the lipid composition of frozen tissue revealed significantly higher levels of phosphatidic acid, and diminished levels of phosphatidylcholine, phosphatidylethanolamine, and N-acylphosphatidylethanolamine from the control. Regardless of freezing temperature, phosphatidic acid levels increased from 4.7 mole% to nearly 50 mole% of the total phospholipid when frozen tissues were stored 10 days at -20 C. During the same period, N-acylphosphatidylethanolamine decreased from 54.1 mole% to 6.6 mole% phospholipid. At least 8 mole% of the phosphatidic acid increase occurred during slow thawing of the frozen tissues. In autoclaved samples, phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, and N-acylphosphatidylethanolamine levels were not different from the control. Labeling of the lipid-glycerol with (3)H, and fatty acids with (14)C, demonstrated the degradation product was primarily phosphatidic acid. Apparently enzymic destruction of the phospholipids occurred during freezing, cold storage, and thawing.