A transforming growth factor beta 2 (TGF-beta 2)-like immunosuppressive factor in amniotic fluid and localization of TGF-beta 2 mRNA in the pregnant uterus.

This report describes a murine amniotic fluid (MAF) immunosuppressive factor that has properties similar to transforming growth factor beta (TGF-beta). The MAF factor exhibits TGF-beta-like activity in stimulating soft agar colony formation by AKR-2B cells and inhibiting thymidine uptake by Mv1Lu cells. We demonstrate that both the immunosuppressive and TGF-beta-like activities of the MAF factor are completely neutralized by anti-TGF-beta 2-specific antibodies and not by anti-TGF-beta 1-specific antisera. The immunosuppressive factor in MAF is novel in that it appears to be identical or very closely related to TGF-beta 2 and is active in its native state. This active and anti-TGF-beta 2-neutralizable factor chromatographs at approximately 70 kD on Sephadex at neutral pH and appears to be able to complex with alpha-fetoprotein in native amniotic fluid. Chromatography of native MAF under acidic conditions demonstrates a lower molecular mass protein that chromatographs on BioGel in the same position as the mature 25-kD TGF-beta. This protein has the biological properties of TGF-beta and is immunosuppressive. Both of these activities are neutralizable with anti-TGF-beta 2 but not with anti-TGF-beta 1 or other antisera. By Northern analysis, we find high levels of TGF-beta 2 mRNA (with little or no TGF-beta 1) in the pregnant uterus that peak around day 15 of gestation and then fall rapidly by day 19 as birth approaches. The TGF-beta 2-like factor could possibly play a role in maternal immunity, in the retention of the fetal allograft, as well as in regulating fetal and neonatal immunological competence.

Differential expression of neural isozymes by human medulloblastomas and gliomas and neuroectodermal cell lines.

For the determination of their possible utility as tumors markers, 2 neural-associated isozymes, neuron-specific enolase [(NSE) EC 4.2.1.11] and creatine kinase BB [(CK-BB) EC 2.7.3.2], were quantitated by radioimmunoassay in human neuroectodermal-derived cell lines, primary brain tumors, and sera and cerebrospinal fluid (CSF) from brain tumor patients. The NSE content of neuroblastoma cell lines was more than sixfold that of the glioma and medulloblastoma lines; the CK-BB content of neuroblastoma and medulloblastoma lines was fourfold to nineteen-fold that of the glioma and other lines. Expression of NSE in neuroblastoma cell lines was not related to time in culture and was cell line specific. NSE in ex vivo medulloblastomas was raised six to ten times that in astrocytomas and gliomas, although no significant differences were noted for the CK-BB content. Serum and CSF NSE levels were markedly raised above control values in 10 of 29 and 6 of 10 cases of astrocytoma, respectively. Raised CK-BB levels in serum (greater than 10 ng/ml) and CSF (greater than 12 ng/ml) were found in 9 of 18 and 2 of 10 patients, respectively. These data suggest that NSE is preferentially expressed by neuroblastoma lines and medulloblastomas and that NSE and CK-BB may have clinical utility as markers for prognosis, diagnosis, and monitoring of response to therapy.

Estrogenic properties of 3,9-dihydroxy-7,12-dimethylbenz[a]anthracene in rats.

Previously, the 3,9-dihydroxy derivative of benz[a]-anthracene was shown to be weakly estrogenic. The availability of the related diol of the mammary carcinogen dimethylbenz[a]-anthracene, i.e., 3,9-dihydroxy-7,12-dimethylbenz[a]anthracene (3,9-diOHDMBA), prompted a similar study of its estrogenic properties. The competitive binding studies of 3,9-diOHDMBA with 17beta-estradiol in the uterine cytosol of immature SD rats gave a Ka of 1.7 x 10(8) M-1. 17beta-Estradiol (10(-9) M) binding to the 8S binding protein was inhibited by 3,9-diOHDMBA at concentrations similar to those of nafoxidine HCl (1 x 10(-5) M). Bioassay demonstrated that the diol possesses 1/4,464 the activity of 17beta-estradiol.

Estrogenic properties of 3,9-dihydroxybenz[a]anthracene, a potential metabolite of benz[a]anthracene.

The estrogen receptor binding properties of 3,9-dihydroxybenz[a]anthracene (3,9-diOHBA) were determined in uterine cytosol of immature Sprague-Dawley rats by competitive binding experiments with [3H]estradiol and sucrose density centrifugation. 3,9-DiOHBA inhibited estradiol binding to the 8S binding protein at a concentration (1.2 X 10(-5) M) approximately equal to that of nafoxidine-HCl (3 X 10(-5) M) required to inhibit estradiol-specific binding, and bioassay for estrogenic activity substantiated this finding.

Effect of Ca2+ and salt on forms of estradiol cytoplasmic receptor in human neoplastic breast tissue.

After incubation with [3H]estradiol low-salt sucrose gradient centrifugation of 250 human breast tumor cytosis demonstrated receptor-bound steroid in both 4 S and 8 S forms. Cytosols prepared from 24 of these receptor-positive tumors were examined in the presence of KCl, Ca2+ and the serine protease inhibitor, phenylmethylsulfonyl fluoride. After incubation with KCl (0.4 M), interconversion of 8 S and 4 S forms was found in one-half of the 24 cytosols examined whose original low-salt patterns contained well-defined 4 S and 8 S peaks. When cytosols were made 4 mM in Ca2+ 15 to 30 min before the addition of KCl, an additional 4.5 S form was observed in low-salt gradients in 18 tumors, and, in 6 tumors, inactivation of receptor occurred. The effects were not produced when Ca2+ and salt were added simultaneously. After 2 to 24 hr of preincubation with Ca2+ of the same cytosols under the same conditions, an increased recovery of the approximately 4 S receptor complex was observed in low-salt gradient. These Ca2+ effects were inhibited if phenyl-methylsulfonyl fluoride and Ca2+ were added simultaneously to cytosols prior to the addition of salt. The data suggest that human mammary tumor cytosols may contain proteolytic activities that can be activated by Ca2+ and that can effect irreversible changes in the salt-dissociated steroid receptor proteins which can be detected in sucrose density gradients.

P-glycoprotein expression in human breast cancer cells.

Multidrug resistance in Chinese hamster ovary cells is associated with the Mr 170,000 surface glycoprotein. Using our monoclonal antibody to this protein, we have isolated a complementary DNA clone from an expression vector library. This complementary DNA recognizes a 4.5-kilobase mRNA in drug-resistant but not-sensitive Chinese hamster ovary cells; it also recognizes a 5.0-kilobase mRNA in our Adriamycin-resistant subline of the MDA-231 human breast cancer cell line which is not expressed in the drug-sensitive parent line. Southern blot analysis shows that the P-glycoprotein sequences are greatly amplified in resistant Chinese hamster ovary cells but not in the resistant human breast cancer cells, indicating that amplification and expression of the Mr 170,000 P-glycoprotein gene are not necessarily coordinate events. Amplification of this gene may not be required for multidrug resistance in human cells.

Role of transforming growth factor-beta1 in the suppressed allostimulatory function of AIDS patients.

BACKGROUND: The T-cell stimulatory function of accessory cells isolated from peripheral blood lymphocytes of AIDS patients has been reported to be suppressed. These patients also have elevated levels of the immunosuppressive factor transforming growth factor (TGF)-beta1 in their serum and plasma. OBJECTIVE: To explore the role of TGF-beta1 in the loss of accessory cell function of peripheral blood lymphocytes from AIDS patients. METHODS: Fluorescent labeled anti-TGF-beta1 and confocal microscopy were used to detect the presence of TGF-beta1 on the cell membrane of dendritic cells. To assess the role of TGF-beta1 in the inhibition of accessory cell function in AIDS, antibodies against TGF-beta1 or the TGF-beta1 type III receptor, beta-glycan, were added to a mixed lymphocyte reaction. RESULTS: TGF-beta1 was detected on the cell membrane of dendritic cells isolated from AIDS patients. The addition of blocking antibodies against either TGF-beta1 or beta-glycan restored the T-cell stimulatory function to accessory cells from these patients. CONCLUSIONS: T-cell stimulatory function was not irreversibly lost in AIDS patients. Our data suggested that beta-glycan-TGF-beta1 immunosuppressive complexes may contribute to the suppression of accessory cell function in these patients.

Normal and malignant neural cells: a comprehensive survey of human and murine nervous system markers.

Tumor-associated neural markers are finding increased application in diagnostic histopathology and in the development of brain tumor therapy. The major cell-type-specific markers and monoclonal antibodies that identify murine and human neural cells are reviewed in this study. Monoclonal antibodies, raised against fetal and adult neural tissue, neuroectodermal tumor tissue, or cell line immunogens which recognize epitopes on brain tumors are comprehensively described including antigens common to the nervous, hematopoietic, and immune systems. The clinical application of neural cell markers and monoclonal antibodies for the diagnosis, localization, and treatment of neuroectodermal tumors is reviewed.

Antiestrogenic properties of substituted benz[a]anthracene-3,9-diols.

The antiestrogenic potency of benz[a]anthracene-3,9-diol, as well as its 7- and 12-methyl derivatives, was evaluated by measuring he inhibition in the onset of estrus brought about by this compound in ovariectomized rats treated with 17beta-estradiol. At a dose of 0.5 mg and 7,12-dimethyl derivative caused a decrease in the percentage of rats in estrus from 78 to 44%. This decrease is identical with that caused by 0.05 mg of nafoxidine.

Differential regulation of TGF-beta 2 by hormones in rat uterus and mammary gland.

Previous work from this laboratory has shown that transforming growth factor beta 2 (TGF-beta 2) mRNA is abundant in the pregnant uterus. In the present study, we examined the synthesis and secretion of TGF-beta 1,2 and 3 in the rat uterus and mammary gland and show differential secretion and expression of TGF-beta 2 in a tissue specific manner. Elevated levels of TGF-beta 2 were detected in late pregnant maternal plasmas (> 100 pM), and in the milk (> 500 pM) during early lactation. High concentrations of TGF-beta 2 (> 200 pM) were also detected in uterine fluids collected from ovariectomized adult rats after high dose estrogen treatment. TGF-beta 2 mRNA levels were elevated in lobuloalveolar epithelial cells isolated from pregnant mammary gland. Three major transcripts of 3.5, 4.0, and 4.7 kb were seen, of which the 4.7 kb, dominates. Mammary glands of estrogen treated ovariectomized rats showed a similar pattern of TGF-beta 2 transcripts. In contrast, four major TGF-beta 2 mRNA transcripts of 5.7, 4.7, 4.0, and 3.5 kb, with the dominant species of 4.0 and 5.7 kb, were observed in uteri from the estrogen treated animals up to 48 h after the last estrogen injection. This suggests that TGF-beta 2 is regulated in a tissue specific manner. We conclude that the secretion of TGF-beta 2 is tightly regulated by hormones and that estrogen and prolactin are critical factors in the tissue-specific regulation of the local production of TGF-beta 2 in the mammary gland and female reproductive tract.

TGF-beta 2 gene and protein expression in maternal and fetal tissues at various stages of murine development.

The transforming growth factor beta family of peptides have diverse actions on the reproductive tracts of primates and rodents. In this study we report the expression of high levels of mRNA of one member of this superfamily, TGF-beta 2, in the pregnant mouse uterus. Using Northern blot analysis and in situ hybridization techniques, we have examined the pattern of expression of TGF-beta 1, TGF-beta 2 and colony-stimulating factor (CSF-1) in mouse maternal and fetal tissue at specific days of gestation. We report here that TGF-beta 2 is synthesized primarily in maternal decidual and uterine epithelial tissues. We observed a shift in the major site of synthesis from decidua to uterus between days 8.5 and 10.5 of gestation. These data demonstrate that the expression of TGF-beta 2 is differentially regulated in the decidua and uterine epithelial cells at various times during gestation. Small amounts of TGF-beta 2 mRNAs were detected in the fetus, and none was detected in placenta, yolk sac, or amniotic membrane. The uterus is likely the major site of synthesis of the TGF-beta 2 found in mouse amniotic fluid. TGF-beta 1 mRNAs are expressed in the uterus at markedly lower levels when compared to TGF-beta 2 mRNAs in both the decidua and uterus. Our results suggest that there is a unique regulation of TGF-beta 2 during pregnancy which may depend on pregnancy hormone(s) and differentiates it from the other mammalian isoforms of the TGF-beta s. TGF-beta 2 may play an important, albeit unknown, role at the maternal/fetal interface.

Perceptual learning of spatiotemporal events: evidence from an unfamiliar modality.

Perceptual learning was examined in two experiments in which subjects, originally unfamiliar with vibrotactile stimulation, were required to identify dynamic vibrotactile patterns with static visual patterns of the same two-dimensional shapes. In Experiment 1 we examined changes in performance with practice under a variety of vibrotactile spatial and temporal conditions. In Experiment 2 we investigated transfer of learning from one set of vibrotactile patterns to another different set. In neither experiment were subjects supplied with knowledge of results. Substantial perceptual learning (improvement in identification with practice) was observed in Experiment 1, although a minority of subjects did not exhibit improvement. Experiment 2 confirmed the general findings of Experiment 1 and also provided evidence of substantial positive transfer. In both experiments, multidimensional scaling of pattern confusion data revealed that practice (and improvement in identification) did not qualitatively change the relative confusability of patterns, suggesting that the (informative) structure of the patterns, irrespective of familiarity with a specific set of patterns, determined confusability. The findings are interpreted in terms of learning constructs offered by E. J. and J. J. Gibson. We conclude by considering the prospects that a connectionist mechanism can account for the observed perceptual learning.

In search of realistic optimism. Meaning, knowledge, and warm fuzziness.

Is it better to be realistic or optimistic? A realistic outlook improves chances to negotiate the environment successfully, whereas an optimistic outlook places priority on feeling good. But are realistic and optimistic outlooks necessarily in conflict? The author suggests that the fuzzy nature of accuracy typically places only loose boundaries on what it means to be realistic. As a result, there are many forms of optimism that do not, in principle, yield unrealistic assessments. Nevertheless, there remain numerous "optimistic biases" that do involve self-deception, or convincing oneself of desired beliefs without appropriate reality checks. The author describes several ways that realistic and unrealistic optimism can be differentiated and explores the impact of this distinction for current views of optimism. This critique reveals how positive psychology may benefit from a focus on personal meaning and knowledge as they relate to making the most of life.

Framing and conflict: aspiration level contingency, the status quo, and current theories of risky choice.

The effect of positive versus negative frames on risky choice was examined for a variety of scenarios and risks. Preferences in the positive domain were strong and mainly risk averse, with notable exceptions. Preferences in the negative domain, however, were marked by their inconsistency, shown both by an overwhelming lack of significant majority preferences and a surprisingly strong tendency of individual subjects to vacillate in their negatively framed choices across presentations. This finding is accounted for by a proposed aspiration level contingency in which aspiration levels are systematically set to be more difficult to achieve in the face of a perceived loss than a gain. The implications of the results, and the aspiration level contingency, are explored with respect to current theories of risky choice, including Kahneman and Tversky's (1979) prospect theory and Lopes's (1987, 1990) security-potential/aspiration theory.

Training wh-question production in agrammatic aphasia: analysis of argument and adjunct movement.

The present research utilized aspects of the Principles and Parameters Approach (P&PA; Chomsky, 1991, 1993) in linguistic theory as well as findings from the psycholinguistic literature as a basis for examining sentence production in aphasic individuals. We examined the production of particular wh-movement constructions--wh-questions requiring movement of an argument noun phrase (i.e., who and what questions) and those which require adjunct movement (i.e., when and where questions). Using a single-subject experimental treatment paradigm, subjects were sequentially trained to produce these wh-questions and, throughout training, generalization to untrained wh-questions relying on similar wh-movement processes was tested. As well, the influence of training on aspects of narrative and conversational discourse was examined. Seven agrammatic aphasic subjects who evinced difficulty producing (and comprehending) "complex" sentences (e.g., passives, object relative clauses, wh-questions)--sentences that involve movement of noun phrases (NPs) out of their canonical positions, leaving behind a "trace" of that movement or "gap"--participated in the study. Subjects were trained to produce wh-questions by taking them through a series of steps emphasizing the lexical and syntactic properties (e.g., thematic role assignment, movement processes, and proper selection of wh-morpheme) of declarative sentence counterparts of target sentences. Results revealed improved sentence production abilities in all subjects under study in both constrained sentence production and, importantly, in discourse tasks. The argument/adjunct distinction was observed in the sentence production recovery patterns noted in six of the seven subjects. Three of the subjects evinced correct argument movement across trained and untrained question structures when wh-questions relying on argument movement were trained; similarly, for these subjects, training structures relying of adjunct movement resulted in improved adjunct movement. Three of the remaining four subjects who required additional treatment to alleviate their wh-morpheme selection deficits, too showed covariance between argument and adjunct movement structures with each type of movement emerging across structures in temporal sequence. We discuss these data in terms of the operations necessary to produce wh-questions, the importance of considering linguistic and psycholinguistic data when designing treatment programs for language disordered patients, and the contribution that detailed recovery data can make both to understanding the nature of sentence production deficits and to issues regarding normal sentence production.

Moving health care education into the community.

The A+ Asthma Club, an educational program developed for elementary school children in inner-city schools, is offered through a series of six sessions during school hours with an additional three booster sessions. This article describes how the program was designed, its theoretical basis, the curriculum and its staffing.

Do we know what we've learned from listening to the news?

This study investigates the relationship between knowledge acquisition and an awareness of that knowledge within the context of listening to the news. Subjects listened to a recording of a radio news program consisting of regular news items as well as editorials, manipulated to be of high or low personal relevance. They then completed a surprise memory test and rated their confidence in their answers. In contrast to many studies, the results indicated a strong positive confidence-accuracy relationship. Confidence ratings were generally a better predictor of an individual's performance than were predictions based on item difficulty. Whereas subjects reported strong and accurate feelings of knowing, they apparently lacked complementary feelings of not knowing. The implications of these findings and others are discussed.

Effects of NO2 on the response of baboon alveolar macrophages to migration inhibitory factor.

Pulmonary alveolar macrophages (PAM) were obtained by lavage from baboons exposed for 6 mo to 2 ppm NO2 for 8 h/d, 5 d/wk, and the response of these cells to autologous migration inhibitory factor (MIF) was determined. PAM from two of three antigen-sensitized, NO2-exposed animals failed to respond to MIF derived from antigen-stimulated autologous lymphocytes. Similarly, PAM from three of the four NO2-exposed animals had diminished responsiveness to MIF obtained by phytohemagglutinin stimulation of their own lymphocytes. The altered responsiveness resulted from an effect on the macrophages and not on the lymphocytes used to prepare the MIF, as shown by the normal blastogenic responsiveness of the lymphocytes and the normal activity of the MIF thus produced on guinea pig peritoneal macrophages. These results demonstrate that inhalation of 2 ppm NO2 may have important subtle effects on pulmonary cells, which may result in altered immune capabilities within the lung.

Alcoholic mortality: a 12-year follow-up.

This is a study of alcoholic mortality in which time, cause, and age at death were variables of critical interest. Five cohorts of 100 members each were followed 12, 11, 9, 6, and 4 years. A total of 133 cases were located as deceased. The overall case fatality rate (CFR) was .0371. Higher CFR's were observed in years 1 to 6. Cardiovascular disease, violence (homicide, suicide, accidental), cirrhosis, carcionomas, and acute intoxication were the leading causes of death. Violent deaths were more prevalent in younger admission age groups. The cardiovascular/other ratio increased in older admission age groups.