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Fabry disease is an X-linked inherited inborn error of glycosphingolipid catabolism. The deficiency of alpha-galactosidase A leads to the deposition of glycosphingolipids primarily in lysosomes of blood vessel cells. In classically affected hemizygotes clinical manifestations include pain in the extremities, vessel ectasia (angiokeratoma) in skin and mucous membranes, ophthalmological abnormalities, and hypohidrosis. As disease progresses there is renal, cardiac, cerebral and vascular involvement, with most patients experiencing renal insufficiency, cardiac hypertrophy or stroke. Many female carriers of Fabry disease also have symptoms. Recently available enzyme replacement therapy has the potential to control or even reverse disease progression. The present analysis reports on five Austrian families with Fabry disease, cared for by nephrologists in June 2002. Furthermore we discuss potential indications for enzyme replacement therapy in patients maintained on renal replacement therapy.

In this work we study the feasibility of using silver halide sensitized gelatin based on PFG-01 (Slavich) emulsions to construct uniaxial compound lenses. This processing is able to introduce variations in the thickness and refractive index of the emulsion. We prove that these changes are not sufficient to provide the observed variations in Bragg conditions in the reconstruction and that a shear-type effect must exist to explain the performance of processed emulsions. We study the characteristics of a compound lens, obtaining acceptable image quality, good resolution, and the typical field limitation of volume holographic elements.

The purpose of this study was to identify whether experiences of childhood physical and/or sexual victimization would increase women's and men's risk for victimization in adulthood by different perpetrators (any perpetrator regardless of the relationship to the victim; intimate partner perpetrator; non-intimate perpetrator) using a nationally representative sample. Results of hierarchical logistic regression analyses indicated that childhood victimization increased the risk for adulthood victimization by any perpetrator for men and women, and by an intimate partner for women but not men. Female and male victims of physical and/or sexual child abuse are at higher risk for adult victimization by non-intimate perpetrators. These results suggest the appropriateness of interventions among adults or young adults who have been victims of child abuse, to prevent any future victimization in adulthood. To guide the development of such prevention programs, research is needed to identify factors that affect the probability of adulthood victimization among child abuse victims.

Background: Few population-based studies have assessed the physical and mental health consequences of both psychological and physical intimate partner violence (IPV) among women or men victims. This study estimated IPV prevalence by type (physical, sexual, and psychological) and associated physical and mental health consequences among women and men., Methods: The study analyzed data from the National Violence Against Women Survey (NVAWS) of women and men aged 18 to 65. This random-digit-dial telephone survey included questions about violent victimization and health status indicators., Results: A total of 28.9% of 6790 women and 22.9% of 7122 men had experienced physical, sexual, or psychological IPV during their lifetime. Women were significantly more likely than men to experience physical or sexual IPV (relative risk [RR]=2.2, 95% confidence interval [CI]=2.1, 2.4) and abuse of power and control (RR=1.1, 95% CI=1.0, 1.2), but less likely than men to report verbal abuse alone (RR=0.8, 95% CI=0.7, 0.9). For both men and women, physical IPV victimization was associated with increased risk of current poor health; depressive symptoms; substance use; and developing a chronic disease, chronic mental illness, and injury. In general, abuse of power and control was more strongly associated with these health outcomes than was verbal abuse. When physical and psychological IPV scores were both included in logistic regression models, higher psychological IPV scores were more strongly associated with these health outcomes than were physical IPV scores., Conclusions: Both physical and psychological IPV are associated with significant physical and mental health consequences for both male and female victims.

While both men and women can be victims, domestic violence usually consists of assaults on women, and most violence against women occurs within an intimate relationship. In the past twenty years, numerous state and provincial programs to intervene in domestic violence cases have developed. The programs tend to focus on treating batterers, although they also offer counseling to domestic violence victims. The jury remains out on the effectiveness of these programs. A major issue is whether the programs use appropriate standards. After an overview of the prevalence and nature of domestic violence, this article provides a discussion of those standards--their nature, effectiveness, and limitations. Another section discusses use of a batterer intervention program in an urban setting. Yet another section explores the implications of intimate partner violence and looks again at the effectiveness of batterer treatment within intervention programs. The article closes with a look at the way one state addresses domestic violence and treats it as a crime. An inescapable conclusion to be drawn from the discussion is that violence against women has its roots in cultural assumptions that must undergo change if the incidence of that violence is to be reduced.

Background and Objective: Disabling chronic pain is especially devastating among working population and, in many cases, it does not respond to conventional therapies. In chronic pain, the importance of psychosocial and occupational factors, in addition to biological ones, has prompted the development of successful multidisciplinary treatment programmes in various countries. We assessed the outcome of a multidisciplinary therapeutic program for work-disabled selected patients with chronic pain refractory to conventional treatment., Patients and Method: The study included 70 patients (58 women, mean age [SD]: 42 [9]years) with chronic pain and sick leave (mean [SD]: 7 [4] months of work disability) diagnosed with fibromyalgia (51%), chronic low back pain (16%), regional myofascial pain (15%), cervicocraneal syndrome (3%), anquilosing spondylitis (3%), and other conditions(12%). All patients had received previous pharmacological treatment,physical therapy and/or other measures (surgery in 12% cases)without improvement. All patients underwent an intensive multidisciplinary treatment of 4 weeks' duration including medical techniques for pain control, cognitive-behavioural therapy, physical therapy,and occupational therapy. Average follow-up was 10 (4) months(1-24 months) post-discharge., Results: Significant improvements were observed with regard to all relevant variables, as reflected in pre and post-discharge measures: pain(Visual-Analogue Scale 1-10 cm): 7.4 (1.5) versus 3.2 (2) (p <0.01); anxiety (HARS), 19 (7) versus 14 (8) (p < 0.01); depression(BDI), 16 (8) versus 10 (8) (p < 0.01); functional ability(HAQ), 1.6 (0.4) versus 0.6 (0.5) (p < 0.001). At discharge,73% of patients returned to work. In addition, 69% of treated patients maintained the acquired improvement and their employment status at the end of follow-up., Conclusion: Multidisciplinary treatment of chronic pain with special attention to work return is useful for selected patients with a disabling chronic pain syndrome refractory to conventional treatment.

Mutations in the FIC1 gene constitute the molecular defect in familial intrahepatic cholestasis I (Fic1 [Byler's disease]) and benign recurrent intrahepatic cholestasis. This report describes the localization of Fic1 in rat liver and intestine, as well as biochemical and transfection studies that support its function as an energy-dependent aminophospholipid translocase. Immunocytochemistry of rat liver and immunoblotting of membrane fractions localized Fic1 to the canalicular, but not basolateral, plasma membrane domain. In the small intestine, Fic1 was localized to the apical membrane of epithelial cells. The distribution of Fic1 in liver plasma membrane fractions from control and taurocholate-treated rats correlated positively with adenosine triphosphate (ATP)-dependent aminophospholipid (phosphatidyl-serine) translocase activity. In canalicular membrane vesicles, translocase activity had an initial velocity of 3.3 nmol phosphatidylserine (PS) translocated per milligram of protein per minute and a K(m) (ATP) = 1.2 mmol/L; was inhibited by vanadate, N-ethylmaleimide, sodium azide, and calcium; and was unidirectional (i.e., from the outer to the inner canalicular plasma membrane leaflet). Transient transfection of CHOK1 cells with FIC1 cDNA resulted in appearance of FIC1 in membrane preparations and energy-dependent PS translocation in cells. These studies indicate that FIC1 is a canalicular P-type ATPase that participates in maintaining the distribution of aminophospholipids between the inner and outer leaflets of the plasma membrane. How this process produces cholestasis is under study.

An optical Fourier processor that allows the use of broadband light sources and colored inputs is designed, fabricated, and tested. We develop a design technique based on phase manipulation in the Fourier plane to construct an image processor that provides a chromatically corrected image making use of the good aberrations behavior of symmetrical optical systems. Only a small number of diffractive lenses and one achromatic refractive lens are required to obtain a real image. We verify our design experimentally using holographic lenses, which are presented, owing to their versatility, as a good alternative to expensive blazed diffractive elements.

ABC transporter trafficking in rat liver induced by cAMP or taurocholate and [(35)S]methionine metabolic labeling followed by subcellular fractionation were used to identify and characterize intrahepatic pools of ABC transporters. ABC transporter trafficking induced by cAMP or taurocholate is a physiologic response to a temporal demand for increased bile secretion. Administration of cAMP or taurocholate to rats increased amounts of SPGP, MDR1, and MDR2 in the bile canalicular membrane by 3-fold; these effects abated after 6 h and were insensitive to prior treatment of rats with cycloheximide. Half-lives of ABC transporters were 5 days, which suggests cycling of ABC transporters between canalicular membrane and intrahepatic sites before degradation. In vivo [(35)S]methionine labeling of rats followed by immunoprecipitation of (sister of P-glycoprotein) (SPGP) from subcellular liver fractions revealed a steady state distribution after 20 h of SPGP between canalicular membrane and a combined endosomal fraction. After mobilization of transporters from intrahepatic sites with cAMP or taurocholate, a significant increase in the amount of ABC transporters in canalicular membrane vesicles was observed, whereas the decrease in the combined endosomal fraction remained below detection limits in Western blots. This observation is in accordance with relatively large intracellular ABC transporter pools compared with the amount present in the bile canalicular membrane. Furthermore, trafficking of newly synthesized SPGP through intrahepatic sites was accelerated by additional administration of cAMP but not by taurocholate, indicating two distinct intrahepatic pools. Our data indicate that ABC transporters cycle between the bile canaliculus and at least two large intrahepatic ABC transporter pools, one of which is mobilized to the canalicular membrane by cAMP and the other, by taurocholate. In parallel to regulation of other membrane transporters, we propose that the "cAMP-pool" in hepatocytes corresponds to a recycling endosome, whereas recruitment from the "taurocholate-pool" involves a hepatocyte-specific mechanism.

Psychological abuse among battered women has been relatively understudied. However, battered women's reports in the existing qualitative and quantitative research suggest that the effects of psychological abuse can be even more damaging than the effects of physical abuse. The current study attempted to clarify the relationship between psychological abuse and posttraumatic stress disorder (PTSD) within a sample of battered women by statistically controlling for the effects of physical abuse. This study also explored the affective experiences of shame and guilt as important variables in the development of PTSD in battered women. This investigation replicated previous work suggesting that battered women are very much at risk for a diagnosis of PTSD and suggests that clinicians and researchers may need to focus on psychological abuse as a predictor of PTSD symptomatology. The current findings encourage attention to shame reactions in battered women and suggest new directions in the study of PTSD for other traumatized populations.

In the present study we describe a simple and fast method to measure the concentration of total free amino acids in very small amounts of biological tissues. The procedure described here is based on the reaction of free amino acids with o-phthaldialdehyde (OPA) in the presence of a reducing agent, beta-mercaptoethanol (MET), to give a complex which can be measured by fluorescence. It is a very rapid process and has the same reliability as the conventional ninhydrin method of Moore and Stein but is about 500 times more sensitive. The sensitivity of the new protocol is such to permit the determination with high reliability of very small amounts of free amino acids at picomole levels, either in a standard amino acid mixture or in biological tissues, without chromatographic separation of the amino acids. It is particularly useful when the amount of the sample is very low, e.g. on a single pituitary or pineal gland of small animals or on single cells, such as oocytes or eggs, as well as single ganglions or axons of marine invertebrates.

Abdominal scintigraphy shows silent gut inflammation in patients with spondyloarthropathies (Sp) without clinical evidence of gut inflammation. Abdominal scintigraphy images are different than those obtained in patients with ulcerative colitis or Crohn's disease and are not related to the anti-inflammatory drugs administered. The aim of this study was to examine the clinical associations of findings on abdominal scintigraphy in patients with Sp. A total of 204 Sp patients (European Spondylarthropathy Study Group 1991 criteria) and 54 non-Sp controls receiving non-steroidal anti-inflammatory drugs were studied. Abdominal scintigraphy images were obtained at 30 and 120 min after injection of technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO)-labelled leucocytes. 99mTc-HMPAO-labelled leucocyte scans were positive in 104 Sp patients (50.9%) and in six non-Sp controls (2.9%) (P<0.001; OR=8.32; 95% CI=3.23-22.67). Silent gut inflammation was not associated with any of the following: age of onset, duration of evolution, sex, family history of Sp or psoriasis, articular manifestations, extra-articular manifestations, radiological findings or HLA-B27 positivity. Positive abdominal scintigraphy was associated with active disease (P < 0.0001; OR=52.7; 95% CI=19-145.6) and an increase in the C-reactive protein (P < 0.005; OR = 3.4; 95% CI = 1.5-7.4). It is concluded that (a) abdominal scintigraphy using 99mTc-HMPAO-labelled leucocytes is of value in detecting the silent gut inflammation in Sp patients, and (b) silent gut inflammation is related to the clinical activity, but is not associated with any particular type of illness or with HLA-B27.

Newly synthesized canalicular ectoenzymes and a cell adhesion molecule (cCAM105) have been shown to traffic from the Golgi to the basolateral plasma membrane, from where they transcytose to the apical bile canalicular domain. It has been proposed that all canalicular proteins are targeted via this indirect route in hepatocytes. We studied the membrane targeting of rat canalicular proteins by in vivo [(35)S]methionine metabolic labeling followed by preparation of highly purified Golgi membranes and canalicular (CMVs) and sinusoidal/basolateral (SMVs) membrane vesicles and subsequent immunoprecipitation. In particular, we compared membrane targeting of newly synthesized canalicular ABC (ATP-binding cassette) transporters MDR1, MDR2, and SPGP (sister of P-glycoprotein) with that of cCAM105. Significant differences were observed in metabolic pulse-chase labeling experiments with regard to membrane targeting of these apical proteins. After a chase time of 15 min, cCAM105 appeared exclusively in SMVs, peaked at 1 h, and progressively declined thereafter. In CMVs, cCAM105 was first detected after 1 h and subsequently increased for 3 h. This findings confirm the transcytotic targeting of cCAM105 reported in earlier studies. In contrast, at no time point investigated were MDR1, MDR2, and SPGP detected in SMVs. In CMVs, MDR1 and MDR2 appeared after 30 min, whereas SPGP appeared after 2 h of labeling. In Golgi membranes, each of the ABC transporters peaked at 30 min and was virtually absent thereafter. These data suggest rapid, direct targeting of newly synthesized MDR1 and MDR2 from the Golgi to the bile canaliculus and transient sequestering of SPGP in an intracellular pool en route from the Golgi to the apical plasma membrane. This study provides biochemical evidence for direct targeting of newly synthesized apical ABC transporters from the Golgi to the bile canaliculus in vivo.

The bile canaliculus contains at least four ATP-binding cassette (ABC) proteins responsible for ATP-dependent transport of bile acids (spgp), nonbile acid organic anions (mrp2), organic cations (mdr1), and phosphatidylcholine (mdr2). Other ABC transporters (including mrp3) have also been partially localized to the canaliculus; however, their function has not been fully delineated. The specific amount and function of spgp and mrp2 in the canalicular membrane increases in response to taurocholate and cAMP. The mechanism involves increased recruitment of spgp and mrp2 from Golgi to the canalicular membrane by a microtubular and PI3 kinase-dependent vesicular trafficking system. Because the effects of taurocholate and cAMP summate, two distinct pathways are proposed. Mdr family members traffic either directly to the apical plasma membrane or, in the case of spgp, through a separate intracellular pool(s); in either case, there is no direct evidence for transcytosis of ABC transporters from Golgi to basolateral plasma membrane and subsequently to the canalicular plasma membrane. Direct transfer from Golgi to apical membrane was demonstrated by in vivo pulse labeling, in vitro membrane localization, and on-line video microscopy in WIFB9 cells that were stably transfected with mdr1-GFP. A critical role for 3'-phosphoinositide products of PI3 kinase was demonstrated in the intracellular trafficking of canalicular ABC transporters and for optimal transporter activity within the canalicular membrane. These studies suggest that many intracellular components, including ATP, Ca2+, numerous GTPases, microtubules, cytoplasmic motors, and other unknown factors, are required for physiologic regulation of ABC transporter traffic from Golgi to the canalicular membrane. Defects in this complex system are postulated to produce an "intrahepatic traffic jam" that results in defective ABC transporter function in the canalicular membrane and, consequently, in cholestasis.

Objective: To translate into Spanish a version of the Miller-Rahe Recent Life Change Questionnaire and to adapt it to Venezuelan cultural values., Methods: The Spanish version and cross-cultural adaptation of the Miller-Rahe Recent Life Change Questionnaire was done following recently proposed guidelines to preserve semantic, idiomatic, and conceptual equivalence in translations of health assessment instruments. We performed one or more translations into the new language, as well as back-translation, test-retest reliability, and weighting score for the translated instrument., Results: A Spanish version of the Recent Life Change Questionnaire was obtained. Validity of translations was demonstrated with a significant agreement for the nonliteral translations (kappa value = 0.97, P < 0.05). The conceptual equivalence was demonstrated by significant agreement in the back-translations (kappa value = 0.84, P < 0.05). The translated instrument met acceptable levels of reliability, as assessed by Spearman's rank correlation coefficients (> 0.60 for all categories of the questionnaire). The cross-cultural adaptation of the translated instrument required addition and exclusion of items as well as changes of the ranking and scaling of life units in the original questionnaire., Conclusion: A valid and reliable Spanish version of the Miller-Rahe Recent Life Change Questionnaire was produced and adapted to Venezuelan cultural values.

Arrangements are described for the recording of volume holograms with two sections that, when stacked together, work as uniaxial centered lenses and allow one to solve the problem of angular selectivity in the imaging of wide objects. The performance of such systems is examined qualitatively, and suggestions aimed at improving these designs are proposed.

Bile secretion in liver is driven in large part by ATP-binding cassette (ABC)-type proteins that reside in the canalicular membrane and effect ATP-dependent transport of bile acids, phospholipids, and non-bile acid organic anions. Canalicular ABC-type proteins can be classified into two subfamilies based on membrane topology and sequence identity: MDR1, MDR3, and SPGP resemble the multidrug resistance (MDR) P-glycoprotein, whereas MRP2 is similar in structure and sequence to the multidrug resistance protein MRP1 and transports similar substrates. We now report the isolation of the rMRP3 gene from rat liver, which codes for a protein 1522 amino acids in length that exhibits extensive sequence similarity with MRP1 and MRP2. Northern blot analyses indicate that rMRP3 is expressed in lung and intestine of Sprague-Dawley rats as well as in liver of Eisai hyperbilirubinemic rats and TR- mutant rats, which are deficient in MRP2 expression. rMRP3 expression is also transiently induced in liver shortly after birth and during obstructive cholestasis. Antibodies raised against MRP3 recognize a polypeptide of 190-200 kDa, which is reduced in size to 155-165 kDa after treatment with endoglycosidases. Immunoblot analysis and immunoconfocal microscopy indicate that rMRP3 is present in the canalicular membrane, suggesting that it may play a role in bile formation.

Bile acid transport and secretion in hepatocytes require phosphatidylinositol (PI) 3-kinase-dependent recruitment of ATP-dependent transporters to the bile canalicular membrane and are accompanied by increased canalicular PI 3-kinase activity. We report here that the lipid products of PI 3-kinase also regulate ATP-dependent transport of taurocholate and dinitrophenyl-glutathione directly in canalicular membranes. ATP-dependent transport of taurocholate and dinitrophenyl-glutathione in isolated canalicular vesicles from rat liver was reduced 50-70% by PI 3-kinase inhibitors, wortmannin, and LY294002, at concentrations that are specific for Type I PI 3-kinase. Inhibition was reversed by addition of lipid products of PI 3-kinase (PI 3,4-bisphosphate and, to a lesser extent, PI 3-phosphate and PI 3,4,5-trisphosphate) but not by PI 4, 5-bisphosphate. A membrane-permeant synthetic 10-mer peptide that binds polyphosphoinositides and leads to activation of PI 3-kinase in macrophages doubled PI 3-kinase activity in canalicular membrane vesicles and enhanced taurocholate and dinitrophenyl-glutathione transport in canalicular membrane vesicles above maximal ATP-dependent transport. The effect of the peptide was blocked by wortmannin and LY294002. PI 3-kinase activity was also necessary for function of the transporters in vivo. ATP-dependent transport of taurocholate and PI 3-kinase activity were reduced in canalicular membrane vesicles isolated from rat liver that had been perfused with taurocholate and wortmannin. PI 3,4-bisphosphate enhanced ATP-dependent transport of taurocholate in these vesicles above control levels. Our results indicate that PI 3-kinase lipid products are necessary in vivo and in vitro for maximal ATP-dependent transport of bile acid and nonbile acid organic anions across the canalicular membrane. Our results demonstrate regulation of membrane ATP binding cassette transporters by PI 3-kinase lipid products.

The contribution of psychological abuse, beyond that of physical abuse, to battered women's psychological adjustment and their intentions to terminate their abusive relationships was examined. Sixty-eight battered women residing in shelters for battered women provided information on their: (1) physical and psychological abuse; (2) psychological symptomatology; (3) strategies for coping with and perceptions of control over partner violence; and (4) intentions to return to their abusive partners. Multiple regression analyses indicated that frequency and severity of physical abuse was not a significant predictor of posttraumatic stress disorder (PTSD) symptomatology nor of women's intentions to terminate their abusive relationships. However, psychological abuse was a significant predictor of both PTSD symptomatology and intentions to permanently leave abusive partners even after controlling for the effects of physical abuse. PTSD symptomatology moderated the relationship between psychological abuse and intentions to terminate the abusive relationships: resolve to leave the abusive partner as a function of level of psychological abuse was significant only among women characterized by low levels of PTSD symptomatology. Greater use of emotion-focused coping strategies, absolutely and relative to problem-focused coping, had direct effects on PTSD symptomatology. However, neither coping nor perceptions of control moderated the effects of psychological abuse on psychological adjustment. The results of the investigation suggested that psychological abuse and ensuing PTSD symptomatology are important variables to assess among physically battered women.

This study investigated lifetime prevalence of traumatic events and posttraumatic stress disorder (PTSD) symptoms among 937 college students. Participants rated their lifetime experiences of traumatic events and, in response to their "most stressful" event, completed measures of objective stressor dimensions, PTSD, and peritraumatic reactions. Approximately 67% of respondents reported at least one traumatic event. An estimated 4% of the full sample (12% of traumatized individuals) met PTSD criteria within the past week. After controlling for vulnerability factors and objective characteristics, peritraumatic reactions remained strongly predictive of PTSD symptoms. Results are discussed with respect to immediate reactions to traumatic events as potential precursors of PTSD symptomatology.

Sjögren's syndrome is an autoimmune exocrinopathy of unknown etiology. It is characterized by a chronic inflammatory process that leads to functional impairment and destruction of lachrymal and salivary glands. There is a primary and a secondary form of the disease; the latter accompanies a well defined connective tissue disease. As far as we know, there are no previous reports of primary Sjögren's syndrome in Venezuela. The main purpose of this study was to present the clinical findings and outcome of a population of patients diagnosed and followed in our hospital. A population of fifty-four patients predominantly females (96%) with a mean age of 42 (range 24 to 84) was studied. The presence of articular symptoms was the most common extraglandular manifestation (87%), followed by enlargement of parotideal glands (52%). Parotid enlargement and renal disease were observed with a higher frequency than previously reported in the literature. Recurrent enlargement of parotideal glands has been related to lymphoid malignant transformation in these patients. Our findings seem to suggest that the pattern of clinical expression of primary Sjögren's syndrome may be influenced by the genetic make-up of the population under study, and possibly by local environmental influences. Two of our cases developed pseudolymphoma, a transitional stage between the benign lymphoproliferation of primary Sjögren's syndrome and lymphoma. Furthermore, in this relatively small sample six patients have died during a short follow-up period, suggesting a potentially more aggressive course of the disease in our patients.

Background & Aims: Guanosine 3',5'-cyclic monophosphate (cGMP), whose production is stimulated by the interaction of nitric oxide, natriuretic peptides, and guanylin with their respective guanylate cyclases, activates secretion through ion channels in several epithelia. Cl- channels have been identified in the apical membrane of biliary epithelial cells. The aim of this study was to investigate the production of cGMP and its effects on Cl- permeability in biliary epithelial cells., Methods: Halide efflux measurement, whole-cell patch clamp recording, radioimmunoassay, and reverse-transcription polymerase chain reaction using two human biliary cell lines (H69 and Mz-ChA-1) were performed., Results: In cells equilibrated with 125I, bromo-cGMP stimulated halide efflux by 22%. In whole-cell patch clamp recordings, the addition of cGMP intracellularly, or of atrial natriuretic peptide extracellularly, stimulated inward currents at negative membrane potentials, consistent with Cl- efflux through open channels. In H69 cells, atrial and C-type natriuretic peptides stimulated production of cGMP. Mz-ChA-1 responded only to atrial natriuretic peptide. Both cell lines expressed messenger RNA for the guanylate cyclase type A receptor and the guanylate cyclase free-clearance receptor., Conclusions: These data suggest that natriuretic peptide stimulates cGMP production in human biliary epithelial cells, which in turn may regulate ductular bile formation through the opening of Cl- channels.

Previous research suggests that certain types of self-appraisals may predispose individuals to be more or less tolerant of relationship violence. The current study investigates two such appraisals, self-esteem and self-attributions, as correlates of women's responses to hypothetical episodes of relationship violence by their dating partners. Undergraduate women involved in dating relationships (N = 145) reported global self-esteem, attributions for hypothetical partner aggression, and probable responses to the aggression. Results showed that self-esteem and self-attributions emerged as correlates of intentions to forgive violence, whereas only self-attributions emerged as a correlate of intentions to dissolve the relationship. The association between self-attributions and intentions to exit a violent relationship was fully mediated by intentions to forgive the partner. Because self-appraisals may inform prevention programs for women who may experience relationship violence, clinical implications are discussed.

Ectonucleotidases are enzymes that degrade extracellular nucleotides. Extracellular nucleotides (especially ATP) and their degradation products (particularly adenosine) have multiple effects on cell functions by acting through purinergic receptors. Adenosine nucleotides are present in bile, which suggests that hepatocytes may release nucleotides into the canaliculus where they are promptly degraded into adenosine by ecto-ATPase and 5'-nucleotidase, which have been identified in the canalicular plasma membrane. Adenosine is then transported into hepatocytes by a Na+-dependent nucleoside transporter that is present in the canalicular plasma membrane. Purification and molecular cloning of ecto-ATPase and other canalicular proteins are complicated by an abundant canalicular plasma membrane protein, cCAM 105. However, the recent cloning of an ecto-ATPase (apyrase) from potato tubers provides a new opportunity to identify the canalicular ecto-ATPase. The canalicular Na+-dependent purine nucleoside transporter has been cloned from rat liver. Study of its expression during development and other physiological circumstances suggests that the transporter may play an important role in maintaining hepatic purine levels that are essential for the liver to serve as a major source of purines for tissues (i.e., brain, muscle) that lack pathways for de novo purine biosynthesis.

Liver endothelial cells possess fenestrae, which are pores supported by a cytoskeleton ring composed of actin and myosin. Fenestrae are dynamic structures that can contract or dilate, although the mechanism for this phenomenon remains to be elucidated. Staining of actin and/or of myosin permitted measurement of fenestral diameter and area in cultured rat liver endothelial cells using digitized video-intensified fluorescence microscopy with image analysis. Within 1 minute of incubation with 0.1 micromol/L serotonin, fenestral diameter and area decreased by 24 +/- 5% and 56 +/- 7%, respectively. Contraction of fenestrae by serotonin was inhibited by chelation of extracellular Ca2+ with EGTA and by addition of Ca2+ channel blockers, such as dilthiazem and verapamil. The response of fenestrae to serotonin was mimicked by addition of a Ca2+ ionophore, A23187. Serotonin inhibited cAMP production, had no effect on inositol phosphate production, and activated phospholipase A2, causing release of arachidonic acid. These results suggest that contraction of fenestrae is associated with Ca2+ influx. In response to 0.1 micromol/serotonin, intracellular Ca2+ levels increased within 3 to 5 seconds from 150 nmol/L to >400 nmol/l followed by rapid phosphorylation of the 20-kd subunit of myosin light chain; both events dependent on extracellular Ca2+.

A murine model in which to study multiple drug resistance in human hepatocellular carcinoma was developed. PRF/PLC/5 hepatoma cells (Alex 0) and an induced multidrug resistant clone (Alex 0.5) were injected intrasplenically into severe combined immunodeficiency mice. In 70% of injected mice, hepatoma cells engrafted in the liver and grew as intrahepatic metastasis. Since Alex cells contain an integrated hepatitis B virus genome and secrete hepatitis B surface antigen (HBsAg), the serum HBsAg concentration in tumor-bearing mice was used to quantitate tumor burden. Tumor wet weight determined at necropsy was directly proportional to the serum HBsAg concentration. In Alex 0 cells, IC50s for doxorubicin, vinblastine, and cis-platinum were 0.35 microM, 0.029 microM, and 3.70 microM, respectively. Alex 0.5 cells were 25-, 14-, and 1.4-fold more resistant to doxorubicin, vinblastine, and cis-platinum, respectively. Immunoblotting of Alex 0 cell membranes with an anti-P-glycoprotein antibody (C219) revealed small amounts of P-glycoprotein, whereas Alex 0.5 membranes overexpressed the protein. Concurrent exposure to verapamil (10 microM) sensitized both cell lines to the cytotoxic action of vinblastine and doxorubicin but had no effect on the cytotoxicity of cis-platinum. Mice bearing intrahepatic xenografts derived from Alex 0 and 0.5 cells had no response to treatment with i.v. vinblastine or doxorubicin, as was anticipated from in vitro drug testing. Addition of verapamil to vinblastine treatment did not improve the success of in vivo chemotherapy. Immunotherapy with a human anti-P-glycoprotein antibody (MRK16) suppressed the in vivo growth of tumors derived from both cell lines. The effect was most pronounced in mice bearing Alex 0.5 tumors. Immunoblotting of tumors which initially responded to MRK16 therapy, but subsequently relapsed, revealed a marked decrease in P-glycoprotein expression when compared to results in tumors that were untreated or treated with vinblastine or control antibody. In summary, we have developed an intrahepatic tumor xenograft model of human hepatocellular carcinoma in mice that permits noninvasive serial quantification of tumor burden by determination of serum HBsAg levels and demonstrated a positive response to immunotherapy with anti-P-glycoprotein antibodies.

The effect of thiol/disulfide exchange on ATP-dependent S-(2,4-dinitrophenyl)glutathione (GS-DNP) transport was studied in sodium nitrate treated rat liver plasma membrane vesicles. Transport followed Michaelis-Menten kinetics with an apparent Km of 9.6 microM for GS-DNP and 124 microM for ATP. 5,5'-Dithiobis(2-nitrobenzoate) (DTNB) and N-ethylmaleimide (NEM) efficiently inactivated GS-DNP transport activity in a dose- and time-dependent manner. Half-maximal inactivation occurred in 10 min at 40 microM for DTNB and 550 microM for NEM. Inactivation by DTNB was reversed by dithiothreitol. S-(N-Ethyl)maleimyl glutathione and/or ATP-Mg2+, but neither S-(N-ethyl)maleimyl cysteinylglycine nor oxidized glutathione could protect transport activity from inactivation by NEM or cystamine. These results suggest that reactive thiols are located near the active site of the transporter and that S-alkyl and the gamma-glutamyl residues of glutathione are important for protection. Biological disulfides which were tested included cystine, oxidized glutathione, oxidized Coenzyme-A, oxidized lipoic acid, and oxidized lipoamide; cystamine was the most potent reversible inactivator. Molecular oxygen also inactivated transport activity, which was recovered on addition of dithiothreitol, suggesting intramolecular disulfide formation by vicinal thiols. We interpret these results to indicate that the ATP-dependent GS-DNP transporter contains two or more thiols which are necessary for the maintenance of transport activity. The reversible inactivation of the activity by biological disulfides suggests that the transporter may be regulated by thiol/disulfide exchange in vivo.

Nonlinear chirped pulse compression can be theoretically achieved to any order by using a nonplane grating with adequate groove spacing. We evaluate the holographic recording of a grating that compensates to the quadratic chirp. A suitable design is found, and the building tolerances are analyzed.

For both theoretical and practical reasons, it is important to understand processes that lead to marital dissatisfaction and dissolution among women who are targets of relationship violence. Because attributional tendencies may often forecast marital behavior and because alcohol use is often seen as providing an excuse for deviant behavior, we examine two potential moderators of the associations between husband violence and wife marital outcomes: wife attributional style and husband problem drinking tendencies. A community sample of married couples (N = 66) completed a comprehensive battery of marital assessments. Results suggested that responsibility attributions moderated the association between husband violence and wives' marital dissatisfaction but exerted a direct effect on wives' disposition toward divorce. Husband problem drinking moderated the impact of husband violence only on wives' disposition toward divorce. As would be expected from an "excuse" model of the associations between violence and marital outcomes, violence had less of an impact on marital satisfaction and divorce ideation when wives attributed responsibility for negative spouse behavior as external to their husbands and when husbands were problem drinkers, respectively.

The adenosine triphosphate (ATP)-dependent transport system is a major determinant of canalicular bile acid secretion. The system transports bile acids and neither organic cations nor non-bile acid organic anions, such as glucuronides or glutathione adducts. To define the structural specificity of the ATP-dependent system, the authors examined the ability of various bile acids to inhibit ATP-dependent taurocholate transport by rat liver canalicular membrane vesicles. Only bile acids with a negative charge inhibited transport, which was unaffected by side chain length. Conjugated, but not unconjugated, mono- and di-hydroxy bile acids inhibited transport. The presence of 7 alpha- and 12 alpha-hydroxylation also influenced inhibition of ATP-dependent taurocholate transport. Inhibition of transport by bile acids was kinetically competitive. These results suggest that the canalicular ATP-dependent bile acid transport system depends on bile acid side chain charge, conjugation, and hydroxylation.

This study examined the role of perceived control and coping in mediating the relationship between violent and nonviolent negative relationship events and women's experience of distress. Results based on the responses of 48 victims of dating relationship violence and 74 nonvictims indicated that perceived control was negatively related to distress for victims but not nonvictims. While both victims and nonvictims engaged in both problem- and emotion-focused coping, and different patterns of coping emerged for the two groups, appraisals of control were not related to choice of coping strategies for violent or nonviolent negative relationship events. Psychological distress was not significantly predicted by coping strategies or the interaction of control and coping for either type of event of for either group. These results suggest that control appraisals may be particularly important in reducing distress for victims. However, appraisals of perceived control may place them at increased risk for abuse in the long run, as victims are unlikely to be able to control the violence as it escalates in both severity and frequency over time.

Background/aims: The biochemical mechanism of bile canalicular contraction is similar to that of smooth muscle contraction. Contraction follows inositol-1,4,5-trisphosphate (InsP3)-dependent Ca2+ release, which activates actin-myosin interactions. Nitric oxide is a myorelaxant through the actions of 5'-cyclic guanosine monophosphate (cGMP) and is produced in hepatocytes exposed to endotoxin and cytokines. The aim of this study was to investigate the effect of nitric oxide on canalicular contraction and to determine the mechanism by which cGMP interferes with the contractile signal., Methods: The canalicular motility in rat hepatocyte doublets was measured by microscopic image analysis, and intracellular Ca2+ was measured by fluorescence microscopy. cGMP and InsP3 were determined by radio-immunoassay and high-pressure liquid chromatography. Ca2+ release from liver homogenate was measured by filtration and superfusion assays., Results: Compounds that release nitric oxide stimulated hepatocellular production of cGMP and prevented agonist-induced contraction by inhibiting the increase in intracellular Ca2+. The cGMP analogue bromo-cGMP prevented contraction and the increase in Ca2+. Bromo-cGMP marginally decreased InsP3 production. cGMP blocked InsP3-dependent Ca2+ release from internal stores., Conclusions: These findings suggest that nitric oxide interferes with Ca2+ signals by cGMP-mediated inhibition of the InsP3 receptor/Ca2+ channel and that hepatocellular production of nitric oxide may be cholestatic by impairing canalicular motility.

Canine lymphoma is a spontaneous, naturally occurring disease that is a model for non-Hodgkin's lymphoma in humans. Chemotherapy with antineoplastics results in a high rate of remission; however, relapse and clinical drug resistance are usually seen within 8-10 months. The P-glycoprotein product of the mdr gene is thought to function as an ATP-driven membrane drug efflux pump and appears to play an important role in tumor cell resistance. To assess the role of mdr gene products in drug resistance in canine lymphoma, membrane preparations of lymphoma cells from 31 dogs with high- or intermediate-grade lymphoma were subjected to Western blotting for detection of P-glycoprotein. In this study, one of 30 samples taken from dogs prior to receiving chemotherapy expressed detectable levels of P-glycoprotein. P-glycoprotein was also detected in biopsy samples from 3 of 8 dogs that had become resistant to chemotherapy. This pattern of expression is similar to that in human non-Hodgkin's lymphoma. These studies suggest that canine lymphoma is a useful model for studying multidrug resistance.

To study the effect of bile acids on P-glycoprotein-mediated drug transport, we performed experiments using multidrug resistant cells and rat canalicular membrane vesicles. Cellular accumulation and efflux of rhodamine 123 were measured in drug-resistant cells by means of computerized quantitative image analysis and fluorescence microscopy. ATP-dependent [3H]daunomycin transport was studied by means of rapid filtration in canalicular membrane vesicles prepared from normal rats. Doxorubicin-sensitive (PSI-2) and -resistant (PN1A) 3T3 cells and human-derived hepatocellular carcinoma doxorubicin-sensitive and -resistant cells were used. Taurochenodeoxycholate and glycochenodeoxycholate, taurolithocholate and ursodeoxycholate (50 to 200 mumol/L) inhibited rhodamine 123 and [3H]daunomycin transport in multidrug-resistant cells and canalicular membrane vesicles, respectively, whereas taurocholate, taurodeoxycholate and tauroursodeoxycholate did not. Primary and secondary unconjugated bile acids had no effect. These results reveal that taurolithocholate, taurochenodeoxycholate and glycochenodeoxycholate and ursodeoxycholate inhibit P-glycoprotein-mediated drug transport function in multidrug resistant cell lines and in canalicular membrane vesicles. These results suggest possible interaction between P-glycoprotein function and bile acids in cholestasis and after treatment of patients with ursodeoxycholic or chenodeoxycholic acid.

We report the case of a previously healthy girl who developed a grand mal seizure after an inappropriate dose of clobutinol and had a good response to iv diazepam.

This study is aimed to describe the differences in mortality among the 38 wards of the city of Barcelona for the period 1983-89. Mortality data for years 1983 to 1989 came from the death certificates. The indicators used for the 38 city wards and the 10 city districts were: Comparative Mortality Figure, Ratio, Ratio of Potential Years of Life Lost, and Life Expectancy at birth. A descriptive analysis of these indicators by wards and district is offered; a cluster analysis based on these indicators was also performed. Wards from the same district are considered homogeneous for a given indicator, when all ward's values are higher, equal or lower to district average. The ward with the most unfavourable indicators and worst situation was Montjuïc, an exponent of shanty town problems until few years ago and located near the centre. Next in ranking, from worst to best situation, did appear the four wards of the city centre district, the old historical quarter. Other wards with high mortality rates were some peripheral areas, nearly all built recently such as Ciutat Meridiana, Bon Pastor and Zona Franca, and also other old wards, such as Poble Sec. Cluster analysis classified the wards with higher mortality in several different clusters, clearly split from the other more homogeneous wards. Five of 10 districts have been considered as homogeneous. This study has allowed a deeper knowledge of the geographical distribution of mortality in the city of Barcelona, until now analysed by city districts.

Most chronic liver disorders are accompanied morphologically by the deposition of fibrous tissue within the hepatic parenchyma. This fibrotic tissue compromises hepatic function and contributes significantly to hepatic failure. Fibrosis is a dynamic process associated with the continual deposition and resorption of connective tissue. Therapeutic strategies are emerging whereby this dynamic process can be modulated. Since collagen is the major component of the extracellular matrix deposited in hepatic fibrosis, most anti-fibrotic therapies have been directed toward the control of collagen metabolism. After collagen genes are transcribed and translated into precursor procollagen proteins, a number of post-translational modifications that ensure the deposition of structurally sound collagen within the extracellular matrix occur. A number of drugs can specifically modulate collagen biosynthesis at the transcriptional level or at various post-translational stages. These anti-fibrotic drugs include corticosteroids, azathioprine, penicillamine, colchicine, zinc, prostaglandins, cyclosporine, and interferons. The pharmacologic action of these drugs and the clinical role in veterinary and human fibrotic hepatopathies will be discussed.

We report the case of an 11-yr-old boy who developed a non-cardiogenic pulmonary edema following inhalation of chlorine gas at a swimming pool. Rapid clinical improvement was noted after i.v. administration of morphine.

The present investigation examined the association between marital investment, marital satisfaction, and commitment to marriage among physically abused women. We applied an investment model and a social learning model to understanding victimized wives' satisfaction and commitment to stay married. Thirty wives who reported physical abuse and 58 nonabused wives completed measures of marital stability, investment in marital problem solving, and dyadic adjustment. Investment in marital problem solving was assessed by having subjects indicate how much energy that they have put into solving 34 common marital problems and whether their efforts were successful or not successful. Consistent with a social learning model but counter to an investment perspective, correlational and multiple regression analyses for each group revealed that failed investment was significantly related to lower, not greater, commitment. Group differences also emerged. Whereas nonabused wives' commitment was related to their dyadic adjustment abused wives' commitment was related to their level of failed investment. Results are consistent with the notion that women may remain in abusive relationships because of psychological entrapment.