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J L Brown

Publications and source records attributed to J L Brown.

At least 37 records · Page 2Linked to original sources
PubMed2001

PTEN inhibits insulin-stimulated MEK/MAPK activation and cell growth by blocking IRS-1 phosphorylation and IRS-1/Grb-2/Sos complex formation in a breast cancer model.

The tumour suppressor gene PTEN encodes a dual-specificity phosphatase that recognizes protein substrates and phosphatidylinositol-3,4,5-triphosphate. PTEN seems to play multiple roles in tumour suppression and the blockade of phosphoinositide-3-kinase signalling is important for its growth suppressive effects, although precise mechanisms are not fully understood. In this study, we show that PTEN plays a unique role in the insulin-signalling pathway in a breast cancer model. Ectopic expression of wild-type PTEN in MCF-7 epithelial breast cancer cells resulted in universal inhibition of Akt phosphorylation in response to stimulation by diverse growth factors and selective inhibition of MEK/extracellular signal-regulated kinase (ERK) phosphorylation stimulated by insulin or insulin-like growth factor 1 (IGF-1). The latter was accompanied by a decrease in the phosphorylation of insulin receptor substrate 1 (IRS-1) and the association of IRS-1 with Grb2/Sos, without affecting the phosphorylation status of the insulin receptor and Shc, nor Shc/Grb2 complex formation. The MEK inhibitor, PD980059, but not the PI3K inhibitor, wortmannin, abolished the effect of PTEN on insulin-stimulated cell growth. Without addition of insulin, wortmannin reduced PTEN-mediated growth suppression, whereas PD980059 had little effect, suggesting that PTEN suppresses insulin-stimulated cell growth by blocking the mitogen-activated protein kinase (MAPK) pathway. Furthermore, PD980059 treatment led to the downregulation of cyclin D1 and the suppression of cell cycle progression. Our data suggest that PTEN blocks MAPK phosphorylation in response to insulin stimulation by inhibiting the phosphorylation of IRS-1 and IRS-1/Grb2/Sos complex formation, which leads to downregulation of cyclin D1, inhibition of cell cycle progression and suppression of cell growth.

Adaptor Proteins, Signal Transducing↗
PubMed2001

A genome survey indicates a possible susceptibility locus for bipolar disorder on chromosome 22.

Bipolar disorder or manic depressive illness is a major psychiatric disorder that is characterized by fluctuation between two abnormal mood states. Mania is accompanied by symptoms of euphoria, irritability, or excitation, whereas depression is associated with low mood and decreased motivation and energy. The etiology is currently unknown; however, numerous family, twin, and adoption studies have argued for a substantial genetic contribution. We have conducted a genome survey of bipolar disorder using 443 microsatellite markers in a set of 20 families from the general North American population to identify possible susceptibility loci. A maximum logarithm of odds score of 3.8 was obtained at D22S278 on 22q. Positive scores were found spanning a region of nearly 32 centimorgans (cM) on 22q, with a possible secondary peak at D22S419. Six other chromosomal regions yielded suggestive evidence for linkage: 3p21, 3q27, 5p15, 10q, 13q31-q34, and 21q22. The regions on 22q, 13q, and 10q have been implicated in studies of schizophrenia, suggesting the possible presence of susceptibility genes common to both disorders.

Bipolar Disorder↗
PubMed2001

Human and mouse IFN-beta gene therapy exhibits different anti-tumor mechanisms in mouse models.

Previously, we suggested that local human interferon-beta (IFN-beta) gene therapy with replication-defective adenoviral vectors can be an effective cancer treatment. Clinical trials to treat cancers with adenovirus expressing the human IFN-beta gene (IFNB1) has been planned. As a continued effort to explore the mechanisms of action of human IFN-beta gene therapy that can occur in the clinical setting, we tested mouse IFN-beta gene therapy in human xenograft tumors in both ex vivo and in vivo models. Delivery of the mouse IFN-beta gene (Ifnb) caused tumor inhibition; this effect was dependent on the indirect anti-tumor activities of IFN-beta, notably a stimulation of natural killer cells. IFN-beta does not show cross-species activity in its anti-proliferative effect and mouse IFN-beta does not cause as significant an anti-proliferative effect on mouse tumor cells as human IFN-beta causes on human tumor cells. Therefore, we believe that mouse models using either human IFN-beta or mouse IFN-beta gene transfer do not capture all aspects of the action of adenovirus-mediated human IFN-beta gene therapy that may be present in the clinical setting. Due to its multiple mechanisms of action, human IFN-beta gene therapy may be effective in treating human cancers that are either sensitive or resistant to the direct anti-proliferative effect of IFN-beta.

Adenoviridae↗
PubMed2001

Different effects of a single amino acid substitution on three adjacent epitopes in the gp41 C-terminal tail of a neutralizing antibody escape mutant of human immunodeficiency virus type 1.

The envelope protein of human immunodeficiency virus type 1 (HIV-1) comprises the outer gp 120 SU domain and the anchoring gp41 TM domain, and the conventional view is that it has a single transmembrane region with the following C-terminal sequence situated entirely within the virion. However, we have recently proposed that the gp41 C-terminal region comprises three transmembrane regions and an external loop structure. Part of this loop is the peptide 731PRGPDRPEGIEEEGGERDRDRS752 that carries three antibody epitopes, 734PDRPEG739, 740IEEE743, and 746ERDRD750. PDRPEG is not detected in virions but reacts with its cognate MAb (C8) in Western blots, IEEE is a linear and non-neutralizing epitope, and ERDRD is a conformational and neutralizing epitope. Here we show that escape mutants selected with neutralizing ERDRD-specific antibody had a single 732R-->G substitution, 14 residues upstream of the cognate epitope, and no longer bound the selecting antibody. The same amino acid substitution altered epitope PDRPEG in the virion so that it now reacted with MAb C8, but left epitope IEEE unaffected. Introduction of 732R-->G by site-specific mutagenesis into the gp41 of cloned HIV-1 NL4-3 virions allowed them to escape neutralization by ERDRD-specific IgG, and confirms that 732R makes a major contribution to the neutralizing conformation of the 731-752 region of the C-terminal tail of gp41.

Amino Acid Substitution↗
PubMed2001

Is epidural analgesia associated with an improved outcome following open Nissen fundoplication?

Postoperative epidural analgesia is increasingly popular in paediatric practice, although evidence of its benefit is scarce. We performed a retrospective analysis of a series of 104 consecutive open Nissen fundoplications, to determine whether mode of analgesia, epidural (n=65) or opioid infusion (n=39), influenced certain outcome measures, including intensive care utilization, duration of hospital stay, morbidity and mortality. The two groups were similar in terms of demographic characteristics and associated pathologies. Overall, morbidity and mortality (2%) rates were low. Mean duration of hospital stay was significantly greater for the opioid group, compared to those receiving epidural analgesia (13 vs. 8 days, P < 0.05). The number of patients who remained in hospital for more than 7 days was also significantly greater in the opioid group. Accepting the limitations of a retrospective study, these data suggest that epidural analgesia might be associated with an improved outcome following Nissen fundoplication and this merits a prospective study.

Adolescent↗
PubMed2001

Ultrasound and endocrine evaluation of the ovarian cycle and early pregnancy in the Sumatran rhinoceros, Dicerorhinus sumatrensis.

Longitudinal ultrasound and endocrine evaluations were conducted in two adult female Sumatran rhinoceroses (Dicerorhinus sumatrensis) over a period of 12-22 months to learn more about their reproductive physiology. Rectal ultrasonography was conducted to monitor ovarian activity. Blood samples were collected and analysed for progesterone and LH, and faecal samples were analysed for progestin metabolites. One female showed cyclic ovarian activity during the study period, whereas the other female showed no evidence of ovarian activity. The cyclic Sumatran rhinoceros appeared to be an induced ovulator, the first of its kind reported within the Perrisodactyla. Ultrasound examinations of the ovaries revealed the formation of anovulatory haemorrhagic follicles when the animal was not mated. These follicles appeared to undergo varied degrees of luteinization that resulted in irregular faecal progestin profiles. When allowed to mate, the female showed a 21 day reproductive cycle that was reflected in both faecal progestin and serum progesterone profiles. The concentration of serum LH was baseline before mating, increased approximately 30-fold within 1-2 h of intromission and returned to baseline within 22 h. Ovulation occurred within 46 h of copulation. The female conceived three times during the study. Pregnancy was detected using ultrasonography 14-16 days after mating, and the concentration of both serum progesterone and faecal progestins remained high. Early embryogenesis appeared to be similar to that in horses. However, each pregnancy terminated unexpectedly within the first 3 months of gestation. This study demonstrates the important role that basic research and reproductive technology can play in developing a natural breeding programme for an endangered animal in captivity.

Abortion, Spontaneous↗
PubMed2001

Are fluid-based cytologies superior to the conventional Papanicolaou test? A systematic review.

OBJECTIVE: We compared fluid-based cytologies (FBC) with conventional Papanicolaou (Pap) tests (CT) to determine if either is superior. STUDY DESIGN: This was a systematic review of original research reports evaluating both CT and FBC with respect to specimen adequacy, comparison with a reference standard, or both. Two reviewers independently reviewed the titles, abstracts, and full articles to determine inclusion status, with differences resolved by consensus with a third author. Risk differences (RD) between occurrence rates for FBC and CT were used for the specimen adequacy data. Sensitivity and specificity were pooled independently and weighted by the inverse of the variance using a random effects model. DATA SOURCES: Studies published between 1985 and November 1999 were identified from MEDLINE, Best Evidence, EMBASE, Biological Abstracts/RRM, and The Cochrane Library. OUTCOMES MEASURED: Sensitivity, specificity, area under the receiver operating characteristic curve (AuROC), and the proportion of satisfactory, unsatisfactory, and "satisfactory but limited by" test results were measured. RESULTS: There was no significant difference in AuROC (P = .37). FBC specimens were more likely to be satisfactory (RD=0.06; 95% confidence interval [CI], 0.03-0.09) or to have absent endocervical cells (RD=0.06; 95% CI, 0.02-0.10) but had 10% fewer "satisfactory but limited by - other" reports (RD = -0.10; 95% CI, -0.14 to -0.06). There was no difference in unsatisfactory Pap test results. CONCLUSIONS: For most women there is no reason to replace CT with FBC. For women at high risk of cervical cancer or who are screened infrequently, the possible increase in FBC sensitivity may outweigh the potential harms from additional false positives.

Confidence Intervals↗
PubMed2001

Understanding the basic reproductive biology of wild felids by monitoring of faecal steroids.

The ability to track gonadal activity is essential for understanding the fundamentals of reproduction. Faecal steroid metabolite monitoring is a well established tool for evaluating reproductive processes in diverse mammalian species, including felids. Domestic cats were used as a model and injection of radiolabelled oestradiol, progesterone, testosterone and cortisol revealed that > 85% of metabolites were excreted in faeces with a time lag of 12-24 h. Steroids were extracted by boiling faecal material (wet or dry) in 90% aqueous ethanol followed by immunoassay with group-specific antibodies that crossreact with excreted metabolites. This approach was used to illustrate the diversity of oestrous cycle characteristics, gonadal responses to photoperiod and ovulatory sensitivity within the felid taxon. Longitudinal analyses demonstrated that faecal oestradiol increases with observed oestrus, and that the duration of the oestrous cycle varies among felid species. Seasonality in gonadal activity was observed in some species (for example clouded leopard, Pallas' cat), whereas other species (for example margay, cheetah, oncilla) are cyclic all year round. Although cats are considered induced ovulators, non-mating and spontaneous ovulation occurred in some species (for example domestic cat, clouded leopard, lion, leopard, margay) with varying frequency. There was also evidence that suppressed ovarian activity and oestrus occurred in group-housed cats (for example cheetahs). As assisted reproductive techniques, such as artificial insemination, are becoming increasingly important for managing zoo species, steroid metabolite monitoring has been especially useful for examining the efficacy of associated hormonal therapies. Exogenous gonadotrophins used to induce ovulation often caused ovarian hyperstimulation, which resulted in a maternal endocrine environment that differed from that of naturally mated cats. Finally, there is evidence that the adrenal status of animals managed under different husbandry conditions can be assessed non-invasively, thereby enhancing our understanding of how social and environmental factors affect animal well-being and reproductive fitness. In summary, understanding the basic endocrinology of endangered felids generates knowledge that can be used to improve management strategies. Because of its enormous utility and non-invasive nature, faecal hormone monitoring is one of the most powerful tools available in zoo research today.

Acinonyx↗
PubMed2000

Comparative fourier transform infrared and circular dichroism spectroscopic analysis of alpha1-proteinase inhibitor and ovalbumin in aqueous solution.

Alpha1-proteinase inhibitor (alpha1Pi) and ovalbumin are both members of the serpin superfamily. They share about a 30% sequence identity and exhibit great similarity in their three-dimensional structures. However, no apparent functional relationship has been found between the two proteins. Unlike alpha1Pi, ovalbumin shows no inhibitory effect to serine proteases. To see whether or not a conformational factor(s) may contribute to the functional difference, we carried out comparative analysis of the two proteins' secondary structure, thermal stability, and H-D exchange using FT-IR and CD spectroscopy. FT-IR analysis reveals significant differences in the amide I spectral patterns of the two proteins. Upon thermal denaturation, both proteins exhibit a strong low-wavenumber beta-sheet band at 1624 cm(-1) and a weak high-wavenumber beta-sheet band at 1694 cm(-1), indicative of intermolecular aggregate formation. However, the midpoint of the thermal-induced transition of alpha1Pi (approximately 55 degrees C) is 18 degrees C lower than that of ovalbumin (approximately 73 degrees C). The thermal stability analysis provides new insight into the structural changes associated with denaturation. The result of H-D exchange explains some puzzling spectral differences between the two proteins in D2O reported previously.

Circular Dichroism↗
PubMed2000

Chorionic gonadotropin administration in domestic cats causes an abnormal endocrine environment that disrupts oviductal embryo transport.

Fecal steroid analysis was used to investigate relationships between endocrine parameters and embryo characteristics in domestic cats subjected to chorionic gonadotropin stimulation and artificial insemination (AI). In Study 1, normal endocrine patterns were assessed in 12 cycling domestic queens. Fecal estradiol (E) patterns established an anovulatory cycle length of 18.3 +/- 0.4 d with estrus lasting 6.3 +/- 0.3 d. Eight females (67%) exhibited at least one spontaneous ovulation based on sustained increases in fecal progestagens (P). In Study 2, queens were mated during natural estrus (NE, n = 5) or subjected to exogenous i.m. gonadotropin stimulation, 100 IU eCG followed by 75 IU hCG 80 h later, (GS, n = 5). Compared with NE queens, fecal E concentrations were higher (P < 0.05) and remained elevated longer after ovulation induction with hCG. In Study 3, gonadotropin-stimulated queens (n = 7) were artificially inseminated and ovariohysterectomized 160 h after hCG. Ancillary follicles and/or corpora lutea were observed in 5 of 6 (83%) ovulating queens. Both fecal E and number of unovulated follicles observed at ovariohysterectomy were negatively correlated with the percentage of embryos recovered from the uterus (r = -0.91 and r = -0.87, respectively; P < 0.05). In summary, exogenous gonadotropin administration causes an abnormal endocrine environment in domestic cats, likely due to ancillary follicle development. The sustained elevations in estradiol appear to impair oviductal transport of embryos, possibly leading to the reduced fertility typically observed in cats subjected to gonadotropin stimulation and AI.

Animals↗
PubMed2000

Basolateral sorting signals differ in their ability to redirect apical proteins to the basolateral cell surface.

Polarized sorting of membrane proteins in epithelial cells is mediated by cytoplasmic basolateral signals or by apical signals in the transmembrane or exoplasmic domains. Basolateral signals were generally found to be dominant over apical determinants. We have generated chimeric proteins with the cytoplasmic domain of either the asialoglycoprotein receptor H1 or the transferrin receptor, two basolateral proteins, fused to the transmembrane and exoplasmic segments of aminopeptidase N, an apical protein, and analyzed them in Madin-Darby canine kidney cells. Whereas both cytoplasmic sequences induced endocytosis of the chimeras, only that of the transferrin receptor mediated basolateral expression in steady state. The H1 fusion protein, although still largely sorted to the basolateral side in biosynthetic surface transport, was subsequently resorted to the apical cell surface. We tested whether the difference in sorting between trimeric wild-type H1 and the dimeric aminopeptidase chimera was caused by the number of sorting signals presented in the oligomers. Consistent with this hypothesis, the H1 signal was fully functional in a tetrameric fusion protein with the transmembrane and exoplasmic domains of influenza neuraminidase. The results suggest that basolateral signals per se need not be dominant over apical determinants for steady-state polarity and emphasize an important contribution of the valence of signals in polarized sorting.

Amino Acid Sequence↗
PubMed2000

A generalized fecal glucocorticoid assay for use in a diverse array of nondomestic mammalian and avian species.

Noninvasive fecal glucocorticoid analysis has tremendous potential as a means of assessing stress associated with environmental disturbance in wildlife. However, interspecific variation in excreted glucocorticoid metabolites requires careful selection of the antibody used in their quantification. We compared four antibodies for detecting the major fecal cortisol metabolites in yellow baboons following (3)H cortisol administration, ACTH challenge, and HPLC separation of fecal glucocorticoid metabolites. The most effective antibody (ICN corticosterone RIA; Cat. No. 07-120102) demonstrated relatively high cross-reactivities to the major cortisol metabolites present in feces during peak excretion, following both radiolabel infusion and ACTH challenge. This same antibody also detected increased fecal glucocorticoid metabolites after ACTH administration in the African elephant, black rhinoceros, Roosevelt elk, gerenuk, scimitar-horned oryx, Alaskan sea otter, Malayan sun bear, cheetah, clouded leopard, longtailed macaque, and northern spotted owl. Results suggest that (1) fecal glucocorticoid assays reliably detect endogenous changes in adrenal activity of a diverse array of species and (2) where comparisons were made, the ICN corticosterone antibody generally was superior to other antibodies for measuring glucocorticoid metabolites in feces.

Adrenocorticotropic Hormone↗
PubMed2000

Evaluating the parent-of-origin effect in bipolar affective disorder. Is a more penetrant subtype transmitted paternally?

BACKGROUND: Numerous genetic mechanisms and modes of transmission underlying bipolar affective disorder (BPAD) have been postulated. Recently, the discovery of genomic imprinting and mitochondrial transmission of illness in humans has stimulated study of parent-of-origin effects in the transmission of BPAD. METHODS: We examined a large sample of families from an associated linkage study to search for a possible parent-of-origin effect. Selecting for unilineal families with at least one offspring and/or parent diagnosed with BPAD after structured interview, we conducted three analyses: (1) the rates of illness among mothers and fathers of offspring affected with BPAD; (2) the observed frequency of transmission and rates of illness among maternal and paternal lineages; and (3) the rates of affective illness among offspring of parents affected with BPAD. RESULTS: Our results indicate no significant differences in the rates of illness among mothers and fathers of offspring affected with BPAD. Also, the frequency of transmission and rates of illness among maternal and paternal lineages did not differ significantly. However, the rate of BPAD among the offspring of fathers affected with BPAD was significantly higher than for mothers with the illness. LIMITATIONS: Substantially more women than men, and maternal than paternal relatives were studied - introducing possible gender biases. CONCLUSIONS: These results suggest a possible paternal parent-of-origin effect.

Bipolar Disorder↗
PubMed1999

Long-term trend toward earlier breeding in an American bird: a response to global warming?

In regions with severe winters, global warming may be expected to cause earlier onset of breeding in most animals, yet no documentation of such a trend exists in North America. In a study of marked individuals of the Mexican jay (Aphelocoma ultramarina) in southeastern Arizona, from 1971 to 1998, the mean Julian date of first clutch in the population declined significantly by 10.1 days. The date of the first nest in the population also became earlier, by 10.8 days. These changes were associated with significant trends toward increased monthly minimum temperatures on the study area, traits that are associated with the onset of breeding in this population. Significant trends from 1971 to 1997 toward warmer minimum temperatures in the months before and during the initiation of breeding were observed. These trends parallel changes in minimum temperatures and community composition in a recent study of grassland ecology in the western United States. Together, they suggest that more attention should be given to the possible ecological importance of global change in minimum temperatures.

Animals↗
PubMed1999

Heterogeneity of in vitro radiosensitivity in human bladder cancer cells.

Human bladder cancer is often heterogeneous containing biologically different populations. Radiotherapy plus chemotherapy is the most common treatment for invasive disease. However few studies have investigated the role of heterogeneity in determining radiosensitivity. The radiation sensitivities of a parent human bladder cancer cell line (UCRU-BL-17CL) and nine cloned cell lines derived from it were determined. These cloned cell lines were previously shown to exhibit different biological characteristics when grown in nude mice. Radiation sensitivity was determined using both MTT and clonogenic assays. The radiobiological parameters, alpha,beta, and surviving fractions at 2 Gy and 8 Gy from the linear-quadratic model, were used to assess radiation sensitivity in the statistical analyses. The nine clones differed in radiosensitivity by both assays. By MTT, but not by the clonogenic assay, their radiation sensitivities were relatively consistent within each of the three biological groups (non-tumorigenic, tumorigenic, invasive); invasive clones were more sensitive than those of the non-tumorigenic and the tumorigenic groups for all the three-test criteria. The heterogeneity exhibited by this cell line may explain some of the variations in the clinical responses seen in the radiation treatment of invasive bladder cancer.

Animals↗