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Biomedical subjects

P Fields

Publications and source records attributed to P Fields.

At least 19 recordsLinked to original sources
PubMed2004

Phase I study of an immunomodulatory thalidomide analog, CC-4047, in relapsed or refractory multiple myeloma.

PURPOSE: To assess the safety, efficacy, and immunomodulatory effects of CC-4047 (Actimid; Celgene, San Diego, CA) in patients with relapsed or refractory myeloma. PATIENTS AND METHODS: Twenty-four relapsed or refractory patients were treated with a dose-escalating regimen of oral CC-4047. Clinical responses and adverse effects were identified, and peripheral T-cell subsets, serum cytokines, and proangiogenic factors were evaluated. RESULTS: CC-4047 was tolerated with no serious nonhematologic adverse events. All patients were eligible for analysis. Toxicity criteria during the initial 4 weeks of study were used to define the maximum-tolerated dose (MTD). During this period, one patient withdrew with a deep vein thrombosis (DVT) probably caused by an undiagnosed primary melanoma with lymphadenopathy in the groin, one patient withdrew because of progressive disease (PD), and three patients discontinued with neutropenia. Nineteen of 24 patients continued on treatment beyond 4 weeks to PD or development of a serious adverse event. Three further patients developed a DVT at 4, 9, and 11 months. Treatment resulted in a greater than 25% reduction in paraprotein in 67% of patients, 13 patients (54%) experienced a greater than 50% reduction in paraprotein, and four (17%) of 24 patients entered complete remission. The MTD was 2 mg/d. All patients showed increased CD45RO expression on CD4(+) and CD8(+) cells, with a concomitant decrease in CD45RA(+) cells. CC-4047 treatment was associated with significantly increased serum interleukin (IL)-2 receptor and IL-12 levels, which is consistent with activation of T cells and monocytes and macrophages. CONCLUSION: This study demonstrates the safety and efficacy of CC-4047. The MTD of CC-4047 orally was 2 mg/d. This is the first report demonstrating in vivo T-cell costimulation by this class of compound, supporting a potential role for CC-4047 as an immunostimulatory adjuvant treatment.

Administration, Oral↗
PubMed2000

Regulation of PKC delta expression by estrogen and rat placental lactogen-1 in luteinized rat ovarian granulosa cells.

Protein kinase C (PKC) delta is dramatically upregulated in the corpus luteum in the second half of pregnancy in the rat. To gain insight into the hormonal regulation of PKC delta expression, studies were undertaken to analyze the regulation of PKC delta expression in a luteinized rat granulosa cell model. PKC delta protein expression was evaluated in luteinized granulosa cells, isolated from human (h)CG-treated immature female rats 7 h after the injection of an ovulatory dose of hCG and cultured up to 12 days. Cytochrome P450 cholesterol side chain cleavage enzyme expression was observed throughout the culture period, and a majority of the cells expressed steroidogenic acute regulatory protein and responded to rat placental lactogen (rPL)-1 by exhibiting hypertrophy, consistent with maintenance of the luteal phenotype. Both PKC delta protein and mRNA expression increased 3.5-4-fold with time of culture, and PKC delta mRNA expression could be eliminated by treatment of cells with the PKC inhibitor GF109203X. E(2) caused a specific dose- and time-dependent increase in expression of PKC delta protein of twofold, whereas PKC delta mRNA was unaffected by E(2) over a 12-day culture period. Treatment of cells with 500 ng/ml rPL-1 for the final 4 days of a 12-day culture in the absence of E(2) had no effect on PKC delta protein or mRNA expression, while treatment with 500 or 3000 ng/ml rPL-1 in the presence of E(2) significantly enhanced both PKC delta protein and mRNA expression (up to threefold). These results show that two of the major regulators of luteal function in the second half of pregnancy in the rat, E(2) and rPL-1, cooperate to regulate PKC delta expression in luteinized granulosa cells.

Animals↗
PubMed1997

Cancer risks in two large breast cancer families linked to BRCA2 on chromosome 13q12-13.

The penetrance of the BRCA2 gene on chromosome 13q12-13 has been estimated in two large, systematically ascertained, linked families, by use of a maximum-likelihood method to incorporate both cancer-incidence data and 13q marker typings in the families. The cumulative risk of breast cancer in female gene carriers was estimated to be 59.8% by age 50 years (95% confidence interval [95% CI] 25.9%-78.5%) and 79.5% by age 70 years (95% CI 28.9%-97.5%). The cumulative risk of breast cancer in male carriers was estimated to be 6.3% (95% CI 1.4%-25.6%) by age 70 years. There was no evidence of any risk difference between the two families. These results indicate that the lifetime breast cancer risk in BRCA2 carriers, for at least a subset of mutations, is comparable to that for BRCA1. A significant excess of ovarian cancer in gene carriers was observed (relative risk 17.69, based on three cases), but the absolute risk of ovarian cancer was less than that reported for BRCA1. Significant excesses of laryngeal cancer (relative risk 7.67, based on two possible carriers) and prostate cancer (relative risk 2.89, based on five possible carriers) were also observed. One case of ocular melanoma, as well as a second eye cancer of unspecified histology, occurred in obligate gene carriers.

BRCA2 Protein↗
PubMed1997

Amino acid sequence differences cannot fully explain interspecific variation in thermal sensitivities of gobiid fish A4-lactate dehydrogenases (A4-LDHs)

We compared the deduced amino acid sequences, heat stabilities and thermal sensitivities of a kinetic property, the apparent Michaelis­Menten constant (Km) of pyruvate, of A4-lactate dehydrogenase (A4-LDH) in four species of goby fishes (Family Gobiidae), adapted to different temperatures, to examine how changes in primary structure influence the adaptation of enzymes. The effect of temperature on Km of pyruvate reflected each species' environmental temperature. For the most eurythermal species, Gillichthys seta, which is endemic to shallow intertidal regions of the upper Gulf of California and encounters temperatures between approximately 9 and 40 °C, Km of pyruvate was minimally affected by temperature, compared with the A4-LDH orthologues from a less eurythermal congener, G. mirabilis (9­30 °C), a cold temperate goby, Coryphopterus nicholsi (10­18 °C) and a tropical species, C. personatus (25­32 °C). Heat denaturation profiles failed to correlate with habitat temperature; G. mirabilis A4-LDH was most thermally stable, followed by the orthologues of C. nicholsi and G. seta. Complementary DNAs (cDNAs) encoding LDH-As of G. seta, Gulf of California and Pacific coast populations of G. mirabilis and C. nicholsi were isolated and sequenced, and the corresponding amino acid sequences deduced. The nucleotide sequences of LDH-A of the two populations of G. mirabilis were identical. Five nucleotide differences in the coding region and one amino acid substitution (at position 78) distinguished LDH-As of G. mirabilis and C. nicholsi. The substitution of a glycyl residue (C. nicholsi) for an alanyl residue (G. mirabilis) may account for the difference in thermal stability between these two orthologues. Comparisons of the LDH-A cDNAs of G. mirabilis and G. seta revealed four differences in nucleotide sequence in the coding region, but all nucleotide substitutions were synonymous. The identical deduced primary structures of the two enzymes suggested the possibility of different protein conformational variants ('conformers') in the two species. This hypothesis is supported by electrospray ionization mass spectrometry, which indicates that the masses of the A4-LDH orthologues of the two species are the same within the resolution of the technique. To explore the possibility that the two enzymes were different conformers of the same primary structure, we treated purified G. seta and G. mirabilis A4-LDHs with 3.0 mol l-1 urea or 6 mol l-1 guanidine­HCl and, after removing the denaturant, compared their kinetic properties and heat stabilities. Neither treatment had an effect on the A4-LDH of G. mirabilis, but both converted the Km versus temperature profile of the G. seta enzyme to that of the G. mirabilis A4-LDH. The thermal stability of neither enzyme was affected. We propose, as has been suggested in several previous studies of A4-LDH, that this enzyme can fold into a number of conformers with different stabilities and functional properties. The A4-LDH of G. seta furnishes evidence that such conformers may provide an important mechanism for adaptation of proteins to temperature.

Journal Article↗
PubMed1996

Control of T lymphocyte signal transduction through clonal anergy.

Stimulation of interleukin-2 producing T lymphocytes via the T cell receptor (TCR) complex in the absence of other costimulatory factor results paradoxically not in activation but in an unresponsive state termed clonal anergy. T cell anergy appears to be a mechanism by which potentially autoreactive T lymphocytes are inactivated in the periphery, thus maintaining tolerance to self antigens. The breakdown of such tolerance may result in autoimmune diseases. In contrast, induction of peripheral tolerance is the ultimate goal in organ transplantation and is a potential mechanism by which a growing tumor evades immune destruction. The anergic state is characterized by an inability to secrete interleukin-2 and proliferate following restimulation via the TCR even in the presence of constimulatory factors. Recent studies have demonstrated a specific block in Ras activation in anergic T lymphocytes. This defect is correlated with a failure to activate the downstream effectors Erk and Jnk and a lack of activation of the AP-1 transcription factor complex, offering a plausible mechanism for the inability to initiate interleukin-2 gene transcription in the anergic state.

Cell Differentiation↗
PubMed1995

Differential requirement for protein tyrosine kinase Fyn in the functional activation of antigen-specific T lymphocyte clones through the TCR or Thy-1.

The protein tyrosine kinase Fyn has been shown to be involved in signal transduction through the TCR and the glycosyl-phosphatidylinositol-linked surface molecule Thy-1 expressed on T cells. In this study, we examine the requirement for Fyn expression in signaling through the TCR or Thy-1 using a panel of Ag-specific T cell clones derived from fyn-/- mutant mice. These clones do not express normal Fyn protein, as measured by immune-complex kinase reaction using anti-Fyn Ab. Stimulation through the TCR, either by APC bearing relevant Ag or by immobilized anti-CD3 mAb, resulted in comparable levels of proliferation, lymphokine production, and cytolysis by clones from both wild-type and fyn-/- mice. In contrast, stimulation through Thy-1, using soluble (or cross-linked) anti-Thy-1 mAb, was deficient, as measured by these responses. Thus, Fyn expression is selectively required for functional activation through Thy-1 in these T cell clones.

Animals↗
PubMed1995

Regulation of T lymphocyte subsets.

Patterns of cytokine secretion and functional differences distinguish T lymphocyte subsets. T lymphocyte subsets are also regulated differentially. Most established CD8+ lymphocyte clones secrete gamma-interferon (IFN-gamma) but not interleukin 2 (IL-2) or IL-4. Using murine T cells which express a transgenic, antigen-specific alpha/beta T cell receptor (TCR) specific for L(d) class I major histocompatibility complex antigen, we have found that CD8+ lymphocytes can be divided into functional subsets. Freshly isolated CD8+ T cells are not cytolytic, do not proliferate and do not proliferate and do not secrete cytokines. Stimulation of TCR alone does not induce cytokine secretion, but cells become responsive to exogenous IL-2 or IL-4. Stimulation of CD28 together with TCR induces secretion of IL-2 and IFN-gamma, and cells proliferate without exogenous cytokines. Proliferation is necessary for the development of cytolytic activity. If IL-4 is present during initial stimulation, IL-4 is secreted following restimulation. Upon stimulation, some IL-4-producing murine CD8+ T cell clones express CD40 ligand (CD40L), and they potentiate proliferation and immunoglobulin secretion by small resting B cells. Thus, the CD8+ T cell subsets T cytotoxic 1 (Tc1) and Tc2 are analogous to CD4+ T helper 1 (Th1) and Th2. IL-2 production by naive CD8+ cells requires co-stimulation. IL-4 production by CD8+ T cells requires the presence of IL-4 during initial stimulation. Some IL-4-producing CD8+ T cells express CD40L following TCR stimulation and provide help for B cells.

Animals↗
PubMed1995

Induction of the increased Fyn kinase activity in anergic T helper type 1 clones requires calcium and protein synthesis and is sensitive to cyclosporin A.

Several alterations in T cell receptor-associated signal transduction have been observed following induction of anergy of T helper type 1 (Th1) clones, including a modified intracellular free calcium ([Ca2+]i) response and increased kinase activity associated with the protein tyrosine kinase p59fyn. In the current study, we demonstrate that, although the kinetics of acquisition of both of these signaling alterations correlated with the generation of anergy, a normal calcium response returned within 48 h after removal from the anergizing stimulus, whereas the increased p59fyn activity persisted and the cells remained hyporesponsive. Generation of both the anergic state and the increased p59fyn activity was prevented in the presence of calcium-free medium, cycloheximide (CHX), or cyclosporin A (CsA), and could be mimicked by the calcium ionophore ionomycin. In contrast, the altered calcium response was inhibited by stimulation in the presence of calcium-free medium or CsA, but not CHX. Thus, surprisingly, these data suggest that a chronic elevation of [Ca2+]i is proximal to and necessary for the increase in p59fyn-associated kinase activity observed in anergic Th1 clones. Increased p59fyn activity, but not the altered calcium response, correlates with maintenance of the anergic state.

Animals↗
PubMed1995

Expression of SQ10 (a preprorelaxin-like gene) in the pregnant rabbit placenta and uterus.

A Northern blot containing poly(A)+ RNA isolated from pregnant rabbit placenta, uterus, ovary, mammary gland, psoas muscle, and intestine was hybridized with an oligonucleotide probe to porcine preprorelaxin. Only the placenta and uterus exhibited high levels of a 1-kb mRNA that hybridized to the probe. Porcine preprorelaxin primers were used to generate and amplify cDNAs from placental and uterine poly(A)+ RNA. Southern blot analysis of the placental and uterine cDNAs each revealed a single PCR product that hybridized with the porcine relaxin probe. The sequence of the cDNAs exhibited 100% identity with SQ10, a preprorelaxin-like gene identified in rabbit tracheobronchial epithelial cells. Identical results were obtained with primers specific for SQ10. SQ10 is expressed in tracheobronchial epithelial cells during abnormal conditions, such as mechanical or toxic injury. The protein is believed to have a role in tracheal epithelial cell differentiation and transformation that occurs under the above conditions. This work shows, for the first time, the presence of high levels of message for SQ10 in normal reproductive tissues. The expression of SQ10 in the uterus and placenta may suggest an important role in cell differentiation and transformation that occurs in these tissues during pregnancy.

Amino Acid Sequence↗
PubMed1995

Induction of lytic pathways in T cell clones derived from wild-type or protein tyrosine kinase Fyn mutant mice.

The OVA-reactive CD4+ Th1 clones and alloreactive CD8+ clones derived from wild-type or fyn-/- mice serve as model systems which have allowed us to investigate several aspects of the molecular events associated with T cell-mediated cytotoxicity, including 1) the differential utilization of two distinct cytolytic pathways by CD4+ Th1 clones and CD8+ CTL, 2) a comparison of the pathways of lysis induced by stimulation of the TCR or by alternative stimuli, 3) the requirement of Fyn for derivation of antigen-specific T-cell clones having properties of CD4+ Th1 and CD8+ CTL cells 4) the differential requirement of Fyn in the induction of responses by TCR and the alternative stimuli. Stimulation through the TCR, either by APC bearing relevant antigen or by immobilized anti-CD3 mAb, resulted in comparable levels of target cell lysis by clones from both wild-type and fyn-/- mice. These clones also utilize the Fas pathway to lyse target cells. Thus, Fyn does not appear to be required for expression of the Fas pathway when triggered through the TCR. In contrast, lysis of target cells by T-cell clones lacking Fyn was deficient when stimulated through Thy-1 or Ly-6C (using mAb) or with Con A or phorbol ester as compared to clones derived from wild-type mice. The basis for the defect in response to stimulation through the GPI-linked molecules appears to be a signaling defect which affects all of the functional responses we measured, while the defect in response to Con A stimulation appears to affect lysis but not lymphokine production. Thus, Fyn expression is selectively required for efficient activation of the Fas pathway of lysis through Thy-1, Ly-6C, and by Con A or phorbol ester in these T-cell clones. CD8+ clones derived from fyn-/- mutant mice, like clones derived from wild-type mice, display antigen-specific lysis, and appear to express perforin message and perforin protein. A Ca(++)-dependent (presumably perforin/exocytosis) component and Fas component of lysis was detected in CD8+ clones derived from fyn-/- mutant mice. Thus, Fyn is not required for expression of these components of antigen specific lysis by CD8+ alloreactive CTL clones. It appears that CD8+ clones that use multiple lytic mechanisms may selectively employ the perforin or Fas-based pathway depending on properties of the target cell or stimulus.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

A 45-year follow-up of kindred 107 and the search for BRCA2.

Interest in the genetics of breast cancer has intensified with the discovery of a breast cancer susceptibility locus, BRCA1, on chromosome 17q. In this paper, we describe updated information on a large breast cancer kindred (K107) that has been extensively studied since 1948. Specifically, we have identified many new cases of cancer in the family and have shown that this family is unlinked to BRCA1 as well as a number of other genes considered as candidates for breast cancer. In a collaborative study between the University of Utah and the Institute of Cancer Research in the United Kingdom, we have collected a set of families with a predisposition to breast and ovarian cancers that have been reliably excluded from linkage to BRCA1 and evaluated their usage in a genomic search for other breast cancer loci. This effort led to the discovery of a second breast cancer locus located on chromosome 13q, BRCA2, which is responsible for the increased incidence of breast cancer in Kindred 107.

Adult↗
PubMed1994

A large kindred with 17q-linked breast and ovarian cancer: genetic, phenotypic, and genealogical analysis.

BACKGROUND: Mutation of a specific, but as yet unidentified, gene BRCA1 on chromosome 17q results in increased susceptibility to breast and ovarian cancer. It is important to know the effects of this gene in terms of the age-specific risks of these cancers and the potential interaction of this gene with other known risk factors. PURPOSE: We performed detailed studies on a large multigenerational family, in which there is known 17q-linked breast and ovarian cancer, in order to characterize the effects of the BRCA1 mutation on development of breast and ovarian cancer. METHODS: Data from the Utah Population Database were used to identify a family (identified as K2082) with a cluster of premenopausal breast cancer and ovarian cancer at any age. Blood samples from 195 members of the family were obtained and these individuals were genotyped for a series of four chromosome 17q polymorphic markers. Information on reproductive history, cancer incidence and treatment, and lifestyle factors was collected on 72 women in the family by questionnaire or through contact with living relatives. RESULTS: Odds in favor of linkage of breast and ovarian cancer in this family to the BRCA1 region of chromosome 17q are greater than 10(8) to 1. The estimated risks for breast or ovarian cancer because of the BRCA1 mutation in this family are 40% by age 50 years and 90% by age 70. No differences between affected and unaffected older BRCA1 gene carriers were observed for a number of known epidemiologic risk factors for these cancers. The gender of the parent from whom the mutant BRCA1 allele was inherited was significantly associated with phenotypic expression (P = .04). A recombinant which places BRCA1 distal to the marker Mfd191 was observed. CONCLUSIONS: Women with the BRCA1 mutation are at increased risk of developing breast and ovarian cancer. In our study population, the mutation appears to confer a lower risk of cancer at younger ages than found in previous studies. Continued interaction with family K2082 will be useful in longitudinal follow-up studies and in studies of the psychosocial implications of providing DNA diagnosis of BRCA1.

Adult↗
PubMed1994

Anergic T-lymphocyte clones have altered inositol phosphate, calcium, and tyrosine kinase signaling pathways.

Full activation of TH1 helper T lymphocytes requires ligation of the specific T-cell antigen receptor (TCR) and a second signal provided by costimulator molecule(s) expressed on particular antigen-presenting cells. Stimulation via the TCR complex alone generates a subsequent unresponsive state characterized by an inability to produce interleukin 2. We report here that such anergic cells exhibit multiple alterations in TCR-associated signaling. The basal levels of intracellular free calcium and phosphatidylinositol 1,4,5-trisphosphate are elevated in anergic cells, and the levels fail to increase significantly upon subsequent restimulation. Examination of phospholipase C-gamma 1 reveals evidence for post-translational modification, correlating with increased tyrosine phosphorylation of the molecule. Tyrosine phosphorylation of additional substrates identified from whole-cell lysates also is altered compared to untreated cells, suggesting a modification in net tyrosine kinase activity. Although the level of kinase activity present in TCR/CD3 or Lck immunoprecipitates is modestly altered after induction of anergy, there is a dramatic increase in specific Fyn-associated tyrosine kinase activity in anergic cells and increased phosphorylation of a 110-kDa protein that is coimmunoprecipitated with Fyn. These results are consistent with a model in which anergic TH1 lymphocytes display a fundamental alteration in TCR-mediated tyrosine kinase activity, associated with changes in phospholipase C-gamma 1, inositol phosphates, and intracellular free calcium.

Animals↗
PubMed1993

Irreversible blindness in systemic lupus erythematosis.

We describe a patient with systemic lupus erythematosis and renal involvement and who developed visual impairment in the absence of hypertension and anti-cardiolipin antibodies. This was attributed to occlusive vascular disease and choroidal infarction in the context of active lupus. Complete and irreversible blindness developed rapidly despite treatment and the patient died of cerebral haemorrhage 2 months after discharge.

Adult↗
PubMed1993

Chromosome 17q linkage studies of 18 Utah breast cancer kindreds.

In this paper we present linkage results from the analysis of 18 Utah breast cancer kindreds, for three 17q markers. Four kindreds had LOD scores greater than 1.0 for at least one of the marker loci. One of these kindreds has a LOD score of 6.07 with D17S579, and we believe it to be the most informative 17q family reported to date. Among the kindreds which appear unlinked to 17q were an early-onset breast cancer family, a large breast-ovarian family, and a kindred with mixed age at onset. Analysis of individual recombinants in the linked families localizes the BRCA1 gene between THRA1 and D17S579 (Mfd188). A comparison of the Cancer and Steroid Hormone Study (CASH) model and a model which assumes a rare dominant susceptibility locus with low penetrance and no phenocopies stresses the difficulties in assessing linkage if the assumptions of the CASH model in terms of age at onset of breast cancer are not appropriate for the BRCA1 locus. A hypothetical breast cancer pedigree is used to calculate gene carrier probabilities under the CASH model, thereby illustrating some of our concerns regarding the use of this model to detect and exclude 17q linkage in breast cancer families.

Adult↗
PubMed1991

An evaluation of antenatal fetal acoustic stimulation.

Fetal heart rate reactivity was evaluated following acoustic stimulation testing (AST). The AST and NST (non-stress test) were compared with regards to certain adverse perinatal outcome criteria, and in a selected sample of cases, to other surveillance tests. These tests were the oxytocin challenge test (OCT) and biophysical profile (BPP). 479 tests were performed on 240 high-risk patients. The AST significantly increased fetal heart rate reactivity. With regards adverse perinatal outcome criteria, there was no significant difference between the AST and NST in relation to sensitivity, specificity or predictive values. In the 38 patients with both non-reactive NSTs and reactive ASTs, 5 (13.5%) had immediate follow-up surveillance tests (OCTs and/or BPPs) that were non-reassuring and necessitated delivery. In four of the five cases, there was evidence of potential fetal compromise. These preliminary observations suggest that the AST may have evoked reactivity in fetuses with early compromise, and raise concerns about replacing the NST with the AST as a primary screening test of fetal well-being.

Acoustic Stimulation↗