The affinity of phospholipase A2 for the interface of the substrate and analogs.

In the intravesicle scooting mode of interfacial catalysis, the interfacial complex E*S is formed by the interaction of the membrane bound phospholipase A2 (E*) with the substrate monomer (S) in the interface. In the presence of nonhydrolyzable substrate analogs (I) the kinetics of interfacial catalysis is modified. If phospholipase A2 is added to a mixture of the vesicles of L-DMPMe ester and of DTPMe ether or D-DMPMe ester, the extent of hydrolysis, A, decreases and the interfacial scooting rate constant, ki, remains unchanged. On the other hand, when the enzyme is added to the vesicles prepared from premixed L-DMPMe ester with D-DMPMe ester or L-DTPMe ether, ki decreases but A remains constant. Qualitatively, these results are in excellent accord with the Scheme I for interfacial catalysis. However, a quantitative departure has been noted, which suggests that the interfacial dissociation constant for E*S is larger than that for E*I. These results are interpreted to suggest that the catalytic rate constant for decomposition of E*S to E* + P is larger than the rate constant for decomposition of E*S to E* + S. Broader implications of the scooting mode of interfacial catalysis are discussed.

Effect of the structure of phospholipid on the kinetics of intravesicle scooting of phospholipase A2.

Action of pig pancreatic phospholipase A2 on vesicles of over 50 synthetic 1,2-diacylglycerol-3-phosphate derivatives and analogs is examined in the absence of any additives. In general, shorter acyl chains and small substituents on the phosphate make a better substrate, while phospholipids with large apolar substituents are not hydrolyzed. The interfacial turnover rate constant for scooting kinetics, ki, for the various phospholipids were from less than 0.1 to 1 per min. Intervesicle exchange of the bound enzyme is faster in vesicles of phospholipids with larger polar substituents, and it is promoted in the presence of anions like chloride, sulfate and thiocyanate. These factors lower the residence time of the enzyme on the bilayer and therefore effectively decrease the rate of hydrolysis. The apparent Km for the enzyme in the interface of anionic phospholipids in the presence of salts is in the 40 to 100 microM range which is 3- to 7-times larger than the dissociation constants for the bound enzyme measured by fluorescence enhancement of Trp-3. The quantum yield of the bound enzyme in vesicles of the various lipids is found to be up to 4-fold different. It is suggested that this difference is due to the E* + S to E*S equilibrium, where E*S has higher fluorescence intensity. The role of calcium in generating the enzyme binding site at the anionic interface, the role of anion anchoring site on the enzyme, and the relationship between the catalytic efficiency and the fluorescence quantum yields are discussed.

Phase transition characteristics of diphosphatidyl-glycerol (cardiolipin) and stereoisomeric phosphatidyldiacylglycerol bilayers. Mono- and divalent metal ion effects.

Synthesis and phase transition chaaracteristics of aqueous dispersions of the homologous (12 : 0, 14 : 0, 16 : 0) diphosphatidylglycerols (cardiolipins) and phosphatidyldiacylglycerols are reported. Electron microscopy of the negatively stained aqueous dispersions reveals a characteristic lamellar structure suggesting that these phospholipid molecules are organized as bilayers in the aqueous dispersions. The phase transition temperature (Tm) and the enthalpy of transition (delta H) increase monotonically with chain length in the cardiolipin and phosphatidyldiacylglycerol series; Tm for phosphatidyldiacylglycerol is higher than that for cardiolipin of the same chain-length. The transition temperatures for the enantiomeric sn-3,3- and sn-1,1-phosphatidyldiacylglycerol and for the diastereomeric, meso-sn-1,3-phosphatidyldiacylglycerol are approximately the same. The molar enthalpy for the transition of cardiolipin-NH+4 bilayers is approximately twice the value for the phosphatidylcholines of the same chain length, i.e., the molar enthalpy per acyl chain is approximately the same in the two systems. The transition temperatures for metal ion salts of C16-cardiolipin exhibit a biphasic dependence upon the unhydrated ionic radii, i.e., the highest Tm is observed for Ca2+-cardiolipin and decreases for the salts of ions with smaller and larger ionic radii than that of Ca2+. The lowest Tm is observed for Rb+-cardiolipin. Monovalent metal salts of cardiolipin exhibit two phase transitions. This effect may result from different conformational packing of the four acyl chains due to differences in metal-phosphate binding.

Phosphatidylcholesterol bilayers. A model for phospholipid-cholesterol interaction.

Aqueous dispersions of monovalent and divalent cations salts of O-(1,2-dipalmitosyl-sn-glycero-3-phosphoryl)cholesterol form multilamellar vesicles as shown by freeze-fracture electron microscopy, by electron micrographs of the negatively stained liposomes, and by swelling curves of liposomes in hypo-osmotic medium. Differential scanning calorimetry reveals that aqueous dispersions of divalent metal salts of O-(1,2-dipalmitoyl-sn-glycero-3-phosphoryl)cholesterol undergo a characteristic thermotropic phase transition with a relatively large cooperative unit (n greater than 250 for the calcium salt). In contrast, monovalent cation salts of O-(1,2-dipalmitoyl-sn-glycero-3-phosphoryl)cholesterol do not show a thermotropic phase transition under comparable conditions. The molecular area of 0-(1,2-dipalmitoyl-sn-glycero-3-phosphoryl)cholesterol in a monolayer is the same in the presence and absence of Ca2+, and is virtually equal to the area of an equimolar mixture of dipalmitoyl phosphatidic acid and cholesterol. To account for the novel state induced by Ca2+, on aqueous dispersions of O-(1,2-dipalmitoyl-sn-glycero-3-phosphoryl)cholesterol (i.e., bilayer organization and highly cooperative phase transition), a linear array model is proposed in which Ca2+ bridges adjacent arrays of O-(1,2-dipalmitoyl-sn-glycero-3-phosphoryl)cholesterol molecules, thus freezing the acyl chains in their normal state. One of the main corollaries of the model is that the cooperative unit for a thermotropic phase transition is essentially one-dimensional, rather than a two-dimensional matrix. O-(1,2-Dipalmitoyl-sn-glycero-3-phosphoryl)cholesterol is proposed as an orientationally and conformationally restricted analog of glycerophospholipid plus cholesterol in bilayers.

Kinetics of interfacial catalysis by phospholipase A2 in intravesicle scooting mode, and heterofusion of anionic and zwitterionic vesicles.

In this and the following three papers we examine the kinetics of action of pig pancreatic phospholipase A2 on vesicles of anionic phospholipids without any additives. The results provide the first unequivocal demonstration of interfacial catalysis in intravesicle scooting mode. In this paper we describe the conditions in which the action of pig pancreatic phospholipase A2 on DMPMe (ester) vesicles in the absence of any additive commences without a latency. Under these conditions the free monomer substrate concentration is insignificant; the bilayer enclosed vesicle organization remains intact even when all the substrate in the outer monolayer has been hydrolyzed; the rate of intervesicle exchange and the rate of transbilayer movement (flip-flop) of molecules is negligibly slow; and the rate of fusion of vesicles is insignificant. Thus an enzyme molecule bound to one vesicle hydrolyzes all the DMPMe molecules in the outer monolayer of the vesicle by a first-order process with a rate constant of 0.6 per min at 30 degrees C; or viewed another way, one enzyme molecule in a DMPMe vesicle can hydrolyze all the available substrate molecules at the rate of 3000 per min. At low anion concentrations excess substrate vesicles are not hydrolyzed unless the rate of intervesicle exchange of the bound enzyme is stimulated by anions in the aqueous phase. Higher calcium concentrations promote not only homofusion of DMPMe vesicles but also heterofusion of DMPMe and DMPC vesicles. It is proposed that calcium-induced isothermal lateral phase separation in DMPMe vesicles induces defects in the bilayer organization, and such defects are the sites for phospholipase A2 binding and for heterofusion with DMPC (ester) vesicles which do not have such sites.

Lateral interaction of cholesterol in diacylphosphatidylcholesterol bilayers.

Thermotropic phase-transition properties of the aqueous dispersions of several diacylphosphatidylcholesterol (DRCh) analogs are examined. The aqueous dispersions of their calcium salts exhibit characteristic endothermic thermotropic transitions due to a change in the conformation of acyl chains. These dispersions consist of osmotically intact liposomes that trap ions, and at the transition temperature there is anomalous increase in the ion leakage. Wide-angle electron diffraction studies of DPCh . Ca monolayers also exhibit a transition from a sharp 4.25 A band to a broad one centering at 4.7 A, reflecting an order-disorder transition in the acyl chains. The long-range order in the organization of acyl chains of DRCh molecules could arise from intermolecular interactions between the cholesterol moieties to form a functional dimer, and such dimers are apparently cross-linked by Ca2+ to form a long-range interacting lattice of acyl chains. Evidence for this model is adduced from the fluorescence properties of the dispersions of dimyristoylphosphatidylcholesta-5,7,9-trienol. The phase-transition properties of DRCh are an ideal illustration of calcium-induced isothermal phase transition.

The lipophilicity, pharmacokinetics, and cellular uptake of different chemically-modified tetracyclines (CMTs).

CMTs are analogs of tetracyclines, which are chemically modified to eliminate their antimicrobial efficacy but which retain their inhibitory activity against matrix metalloproteinases. These compounds have been found to inhibit connective tissue breakdown in animal models of diseases such as periodontitis, arthritis and cancer. Because CMTs exhibit different in vivo efficacy in these various models of disease, the current study compared their pharmacokinetics and other properties as follows: Adult male Sprague-Dawley rats were administered by oral gavage a single dose of 5mg of different CMTs suspended in 1 ml 2% carboxymethyl-cellulose, and blood samples were collected from 1-48 hours after dosing. The sera were extracted, then analyzed by HPLC using a C-18 reverse-phase column. The results showed that the peak concentrations (C(max)) in rat sera 1-12 hours after oral administration of CMTs -1, -2,-3, -4,-5,-6,-7,-8 and doxycycline were 5.5, 0.7, 4.6, 6.2, 0.8, 0.7, 9.0 (note: the 3 peaks detected for CMT-7 were combined), 15.0 and 0.9 microg/ml, respectively. Their in vivo half-lives (t(1/2)) were 11, 5, 22, 11, 32, 15, 37, 38, and 17 hours, respectively. Of the anticollagenase CMTs tested, CMT-8 showed the greatest C(max) and t(1/2)values, followed by CMTs-3, -1, -4, and perhaps -7; CMTs-2, -5, and -6 exhibited much lower levels in serum. The relative lipophilicities of the 8 CMTs and doxycycline were tested by examining their extractability in octanol. The results showed that CMT-2, -5, and -6 had the lowest partition coefficients using this organic solvent, while CMT-3 was the most lipophilic. The lipophilicity of the different CMTs was also positively correlated (r(2)=0.767, P<0.05) to peak serum concentrations (C(max)), but not to their serum half-lives (r(2)=0.25,P=0.49). This property of the different CMTs was also found to be positively correlated to their ability to enter into human whole blood cells in vitro (r2=0.95, P<0.001). Since CMT-8, as well as CMTs-3 and -1, consistently exhibited the greatest in vivo efficacy in animal models of tissue breakdown, this may reflect, at least in part, their favorable pharmacokinetics and tissue uptake.

Controlled chemical modification of hyaluronic acid: synthesis, applications, and biodegradation of hydrazide derivatives.

Controlled modification of the carboxylic acid moieties of hyaluronic acid with mono- and polyfunctional hydrazides leads to biochemical probes, biopolymers with altered physical and chemical properties, tethered drugs for controlled release, and crosslinked hydrogels as biocompatible scaffoldings for tissue engineering. Methods for polyhydrazide synthesis, for prodrug preparation, for hydrogel crosslinking, and for monitoring biodegradation are described.

Synthesis and evaluation of 6-[(18)F]fluoro-3-(2(S)-azetidinylmethoxy)pyridine as a PET tracer for nicotinic acetylcholine receptors.

Both ABT-594 ((R)-2-chloro-5-(2-azetidinylmethoxy)pyridine) and A-85380 (3-[2(S)-2-azetidinylmethoxy]pyridine), novel nicotinic agonists that possess potent non-opioid analgesic properties, have high affinity for neuronal nicotinic acetylcholine receptors (nAChR) but do not elicit the pronounced toxicity of epibatidine. 6-[(18)F]Fluoro-3-(2(S)-azetidinylmethoxy)pyridine (6-[(18)F]fluoro-A-85380), a F-18 labeled analogue of these two compounds, is therefore a promising radioligand for positron emission tomography (PET) studies in humans. The use of trimethylammonium as a leaving group in nucleophilic aromatic substitution reactions has proven to be a versatile and efficient strategy, and offers several advantages over other leaving groups. Here, we report the synthetic strategy for the preparation of a precursor, as a trimethylammonium iodide salt, and its use in the radiosynthesis to 6-[(18)F]fluoro-A-85380. Preliminary compartative PET studies of 6-[(18)F]fluoro-A-85380 and 2-[(18)F]fluoro-A-85380 were carried out in baboon to examine their suitability as tracers for studying nAChR system.

Gender, politics, and psychology's ways of knowing.

Recent work on the psychology of gender has emphasized comparisons of men and women. Such comparisons rest on a view of gender as an individual difference or psychological attribute. Feminist theorists have challenged this view as limited and inadequate. In place of it, a variety of alternative conceptions of gender are emerging. These conceptions shift the focus of analysis from the individual to interpersonal and institutional arenas. Moreover, they dispute the idea of gender as static, unitary, and separable from other markers of social identity and status. In contrast to Alice Eagly (1995), I assert that the production of knowledge (whether by scientific procedures or other means) is not set apart from society, but rather is always and inevitably embedded within it. Therefore, I call for efforts to uncover the ways in which psychological knowledge is shaped by ongoing societal struggles and cultural politics.

Counseling adolescents with problem pregnancies.

PIP: The whole gamut of problems involved in counseling pregnant teens, from the national statistics and psychosocial context to the goals of counseling and follow-up are discussed. In the U.S. 40% of women have become pregnant by age 20 (1982 data), making this one of the highest rates of teenage pregnancy worldwide. Many factors contribute to this problem: adolescent development with its unresolved conflicts; teenage cultural myths about sex roles, such as the idea that it is harmful to deny sex to males; cultural roles for women such as being passive and therefore unprepared with contraception. The teen pregnancy, then, evokes a personal, interpersonal, moral and medical crisis. Goals for counseling are to help the young woman mjake a fully integrated, final decision; provide emotional support and referral information; to help her use the experience for growth. The decision often has to be rapid, because many teens delay revealing their pregnancy out of fear, yet prenatal care is essential, while a second trimester abortion should be avoided. Decisions about whether to bear the child, involve the father, the family, where to live, how to get financial support become complex. Even the counselor's background can be counterproductive, especially when the teen chooses STET unlikely to succeed. Issues of client's autonomy and confidentiality regarding family, the male partner, and even local regulations may confuse the process. Follow-up counseling may be indicated, for post-abortion or pregnancy support, for future contraception, for a committed relationship, or for balancing her needs versus her sexual partner's.^ieng

New tetherable derivatives of myo-inositol 2,4,5- and 1,3,4-trisphosphates.

(+/-)-myo-Inositol 1-(3-aminopropyl hydrogen phosphate) 3,4-bis(disodium phosphate) (5) and (+/-)-myo-inositol 2-(3-aminopropyl hydrogen phosphate) 4,5-bis(disodium phosphate) (11) have been synthesized by conventional procedures. Each derivative has been immobilized on a polymeric resin in order to give a bioaffinity matrix.

Culture, gender, and suicidal behavior in Sri Lanka.

In less than 50 years, the rate of suicide among Sri Lankans has risen from a modest level to one of the highest in the world. This article describes the current pattern of suicides, including sex differences and similarities, and reviews some of the institutional practices, material conditions, and social norms that may figure in the increases. A study is reported, which asked how ordinary Sri Lankans account for suicidal behavior and what practices they recommend for assisting suicidal individuals. Gender inflected these accounts, with essentialist accounts associated with women's suicides and contextual accounts associated with men's suicides.

Women and mental health: a review of feminist change efforts.

A CENTRAL CONCERN of the women's movement has been the damage done to women's psychological well-being by traditional views of women and femininity. This paper will review some of the negative effects of social factors on women's psychological well-being and will discuss four areas in which feminists are striving to change the mental health system.

Dentine lead concentration as a predictor of neuropsychological functioning in inner-city children.

Cumulative lead exposure in 193 inner-city Black children was evaluated by measuring lead concentrations in the primary dentine and the circumpulpal dentine of their deciduous teeth. The children's dentine lead levels were comparable to those reported in population studies of low-income children living in inner-city areas. The results of neuropsychological tests administered at age 7 were available for children in the sample; we performed further neuropsychological assessments when their teeth were shed (10-14 yr of age). Higher dentine lead concentrations were associated with deficits in neuropsychological test performance at both testings, with performance abilities more strongly affected than verbal abilities. Moreover, the negative effects of lead on neuropsychological functioning were more evident when lead concentrations in circumpulpal dentine, rather than in primary dentine, was used as the index of lead exposure. Based on this finding, we suggest that circumpulpal dentine and primary dentine should be assayed separately in order to yield a more sensitive estimate of exposure to lead.

Low-level lead exposure in childhood influences neuropsychological performance.

Cumulative lead exposure in 193 inner-city black children was assessed by measuring lead concentrations in the primary and circumpulpal dentine of their deciduous teeth. Lead concentrations for these children were comparable to the concentrations reported in population studies of low-income children living in inner-city areas. Analysis of the children's neuropsychological test performance showed that elevated lead levels were associated with deficits in visual-motor functioning and perceptual integration, right-left orientation, and verbal abstraction; other verbal abilities and motor functioning were not affected. Possible alternative explanations for these findings were evaluated by examining the relationships between lead and maternal intelligence, family socioeconomic status, and perinatal indicators of neurological impairment. None of these relationships was significant. It was concluded that, even at levels usually regarded as asymptomatic, lead in the environment represents a hazard to inner-city children.

The concentration levels of Cd, Pb, Hg, Cu, Zn and Se in blood of the population in the Czech Republic.

Knowledge of normal levels of concentrations of trace elements (Cd, Pb, Hg, Cu, Zn, and Se) in the population serves, among others, in design of regulations concerning health protection, determination of exposure limits and prevention of diseases caused by deficiency of trace elements. Concentrations of the named elements in whole blood of the Czech population were determined by means of atomic absorption spectrometry. The blood was collected during 1996-1998 from 1,216 blood donors (896 males and 320 females, average age 33 years) and 758 children (397 boys and 361 girls, average age 9.9 years). Mineralisation in a microwave digestion system was used in sample preparation. The accuracy of results was checked by means of the Control material Seronorm Whole Blood 404107 and Seronorm Serum 704121, Nycomed. Values of concentrations of the trace elements in blood found for adult (medians) were 0.7 microgram Cd.l-1, 800 micrograms Cu.l-1, 0.78 microgram Hg.l-1, 41 micrograms Pb.l-1, 76 micrograms Se.l-1, and 5,800 micrograms Zn.l-1, respectively. Statistically significant differences between men and women have been found in the concentrations of Cu, Hg, Pb, and Zn. In the juvenile population following medians of concentrations have been found: 0.15 microgram Cd.l-1, 1,047 micrograms Cu.l-1, 0.46 microgram Hg.l-1, 34 micrograms Pb.l-1, 69 micrograms Se.l-1, and 8,180 micrograms Zn.l-1. Statistically significant differences between boys and girls were found only in Pb and Zn concentrations. Concentrations of the studied elements correspond to the published values concerning population not exposed professionally.

Gender roles in the relationships of lesbians and gay men.

Recent research on gay male and lesbian couples suggests that traditional gender-role-playing sometimes occurs in their relationships, though it is less common than in the relationships of heterosexuals. This paper briefly explores three issues raised by these findings. First, we consider reasons why partners of the same gender might engage in gender-role-playing. Second we discuss the processes that might be involved in allocating masculine and feminine roles to partners in a couple. Finally, we consider the finding that traditional gender roles are associated with diminished satisfaction and suggest possible reasons why this might be so.