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Biomedical subjects

C Miller

Publications and source records attributed to C Miller.

At least 127 records · Page 7Linked to original sources
PubMed1999

Anergy and cytokine-mediated suppression as distinct superantigen-induced tolerance mechanisms in vivo.

Recombinant-activating gene 2 (RAG-2-/-) T cell receptor-transgenic mice repeatedly injected with the superantigen staphylococcal enterotoxin A entered a tolerant state in which splenic CD4+ T cells produced little interleukin (IL)-2, interferon gamma, or IL-4. This state resulted from a combination of both clonal anergy and cytokine-mediated immunosuppression. The anergy persisted for at least 3 wk and could be distinguished from the suppression by a decrease in IL-2 production per cell, a block in the activation of early response kinases, and a failure to be reversed with anti-transforming growth factor (TGF)-beta. Full suppression lasted for only 1 wk and involved both IL-10 and TGF-beta, but required additional unknown molecules for optimal effect. These experiments show that complex in vivo interactions of multiple peripheral tolerance mechanisms can now be dissected at both the cellular and molecular levels.

Animals↗
PubMed1999

Meta-analysis of sentinel lymph node biopsy in breast cancer.

BACKGROUND: Sentinel lymph node biopsy (SLNB) is a minimally invasive way to diagnose axillary lymph node (ALN) metastases in breast cancer. The most important features are ability to identify the SLN (I.D. rate), how often the SLN and ALN pathology match (concordance), and how often the SLN is negative for cancer when the ALNs are positive (false negative). Technique and patient criteria for SLNB vary among studies. This study performed meta-analysis of published studies to determine the I.D., concordance, and false negative rate (1) overall and for (2) both blue dye and radiocolloid, (3) the injection method, (3) palpable and nonpalpable ALNs, and (4) invasive and in situ disease. METHODS: Inclusion criteria were patients with breast cancer who had SLNB followed by ALN dissection with H&E staining. Meta-analysis was performed using analysis of variance with each observation weighted inversely to its variance. P < 0.05 was considered significant. RESULTS: Eleven studies (n = 912) met the inclusion criteria. Overall, 762 (84%) SLNs were identified, concordance was 747/762 (98%), and 15/296 (5%) were falsely negative. Highest I.D. rates (P < 0.05) were reported with albumin radiocolloid or dye + radiocolloid (97 and 94%, respectively), with injection around an intact tumor (96%), with invasive cancer (95%), and in the clinically negative axilla (96%). Concordance and false negative rates did not vary. CONCLUSIONS: The SLN can be identified in over 97% of patients if certain techniques and inclusion criteria are used. SLNB reflects the status of the axilla in 97% of cases and has a 5% false negative rate.

Adult↗
PubMed1999

Introduction to stem cell biology in vitro. Threshold to the future.

Transplantable hematopoietic cells with multilineage reconstituting ability can be quantitated in suspensions of human or murine cells using similar assay procedures. The incorporation into these assays of stringently defined functional endpoints ensures a high degree of specificity for the cells detected. Application of these assays to stem cell-containing suspensions after they have been stimulated for several days with defined cytokines in vitro, or by a mixture of defined and/or undefined factors in vivo, has shown that net amplifications in these populations can be obtained under both circumstances. Such studies have allowed cytokine conditions that support stem cell self-renewal divisions to be identified and have also provided evidence that stem cell regeneration can be manipulated both in vitro and in vivo by altering the molecular milieu of the responding cells. These observations pave the way to future delineation of mechanisms that control the normal behavior, pathology and future clinical exploitation of hematopoietic stem cell populations.

Animals↗
PubMed1999

Administration of oxytocin immediately after insemination does not improve pregnancy rates in mares bred by fertile or subfertile stallions.

It is probable that reduced pregnancy rates in mares bred to subfertile stallions is attributable, in part, to the reduced number of normal spermatozoa that colonize the oviduct. Administration of oxytocin stimulates both uterine and oviductal contractility. The hypothesis that oxytocin may enhance sperm transport to/into the oviducts, and thereby increase pregnancy rates, was tested in 2 trials. For both trials, fertile estrous mares with follicles > or = 35 mm in diameter were inseminated once at 24 h after administration of 1500 to 2000 U hCG. The inseminate dose was limited to 100 million spermatozoa in order to lower pregnancy rates and thus increase the chance of detecting a treatment effect. Pregnancy status was determined by transrectal ultrasound examination 14 to 16 d after insemination. In Trial 1, 49 mares were inseminated with 4 mL extended semen from 1 of 3 stallions (1 fertile and 2 subfertile males). Immediately after insemination, the mares were administered either 20 U oxytocin or 1 mL saline intravenously. In Trial 2, 51 mares were inseminated with 4 mL extended semen from 1 of 4 stallions (1 fertile and 1 subfertile male used in Trial 1, and 2 additional fertile males). Immediately after insemination, and again 30 min later, mares were administered either 5 U oxytocin or 0.25 mL saline intramuscularly. To test for effects of treatment with oxytocin and for the interaction between semen quality and treatment, a generalized linear mixed regression model was used that accounted for the split-plot design (treatment within stallions), the random effect of stallion, the fixed effect of semen quality, the binary outcome of a single breeding trial, and the varying number of trials per stallion/treatment groups. Three treatment protocols or regimens were used: placebo, 5 U oxytocin injected twice intramuscularly, and 20 units oxytocin injected twice intravenously. Semen was classified as high (fertile stallions) or low (subfertile stallions) quality. No interaction between semen quality and treatment was detected (P > 0.10). The pregnancy rate of mares treated with oxytocin immediately after insemination was 30% (15/50) compared with 50% (25/50) for mares treated with saline immediately after breeding. Administration of oxytocin did not affect pregnancy rates (P > 0.10).

Animals↗
PubMed1999

Simultaneous treatment of cytomegalovirus and varicella zoster infections in a renal transplant recipient with ganciclovir: use of viral load to monitor response to treatment.

Disseminated zoster occurring simultaneously with cytomegalovirus (CMV) disease in a renal transplant recipient is potentially life threatening. We describe the use of intravenous ganciclovir to treat both infections. The efficacy of treatment was assessed clinically and by the measurement of CMV viral load using the hybrid capture (Murex version 2) and varicella zoster (VZV) viral load using an in-house assay. Results from this case suggest that clinical resolution in severe viral infections such as described below may be related to early control of viraemia.

Adult↗
PubMed1999

Calphostin C induces AP1 synthesis and AP1-dependent c-jun transactivation in normal human chondrocytes independent of protein kinase C-alpha inhibition: possible role for c-jun N-terminal kinase.

Activator protein-1 (AP1) regulates the promoter activity of a large number of genes associated with developmental, proliferative, inflammatory, and homeostatic processes in human connective tissue cells. Some of these genes (e.g., cyclooxygenase-2) are regulated by the protein kinase C (PKC) inhibitor, calphostin C (CalC). We examined whether CalC could indeed induce AP1 and AP1 gene transactivation (c-jun) in human chondrocytes. Exploratory studies confirmed the anti-PKC effects of CalC, as equal molar concentrations of CalC blocked the PMA-induced translocation of PKC-alpha from the cytosolic to the membrane fraction. CalC induction of AP1, as judged by gel-shift analysis, using a consensus AP1 oligonucleotide, was biphasic with an initial increase (maximum 4 h), followed by a decline, reaching its nadir after 16 h, and finally a major upregulation phase at 24 h. Maximum induction of AP-1 was reached at a concentration of 250 nmol/L of CalC. CalC did not block PMA-induced AP1 synthesis. Gel-shift analysis in the presence of specific antibodies to c-Jun, JunB, JunD, c-Fos, and CREB/ATF showed that the AP1 complexes were probably c-Jun/c-Jun, c-Fos/c-Jun, c-Fos/JunB, or c-Jun/JunB dimers. Northern blot analysis confirmed that c-jun, junB, and c-Fos were the principal proto-oncogenes induced by CalC. To confirm that c-jun induction occurs at the transcriptional level and to examine the role of the AP1 site present in the c-jun promoter in the induction of c-jun by CalC, we performed transient transfections of c-jun promoter-CAT constructs harboring either wild-type (WT) AP1 regulatory element sites or mutant AP1 sites. CalC (250 nmol/L) induced a marked increase in CAT activity (i.e., promoter activation) with WT AP1 c-jun promoter-CAT plasmids, but the response was completely abrogated when using constructs where the AP1 site was mutated. PMA produced similar results, but the induction of the WT AP1 c-jun promoter-CAT plasmid was smaller. CalC (250 nmol/L) inhibited MAPK (p42/44) activity while stimulating c-Jun N-terminal kinase activity in a time-frame coincident with the activation of AP1. We conclude that CalC induces signaling pathways that activate AP1 and transactivate genes harboring AP1 enhancer sites independent of PKC-alpha.

Base Sequence↗
PubMed1999

Perceptions of the effect of an impending restaurant smoking ban on dining-out experience.

BACKGROUND: The introduction of bans on smoking in restaurants is frequently marred by claims that they will lead to a loss of business. METHODS: A representative sample of 3,019 South Australians age 15+ years were asked questions about dining-out frequency and perceived effects of the ban on their dining-out enjoyment and restaurant patronage. RESULTS: Sixty-one percent thought the ban would make dining out more enjoyable, 5% thought it would be less enjoyable, and 34% thought it would make no difference. Overall, 82% thought the ban would make no difference to their likelihood of dining out, 14% would be more likely to dine out, and 4% would be less likely. CONCLUSIONS: These data suggest that the public expects bans on smoking in restaurants to result in both increased enjoyment and increased patronage of restaurants.

Adolescent↗
PubMed1999

Leber's hereditary optic neuropathy mitochondrial DNA mutations in familial multiple sclerosis.

Leber's hereditary optic neuropathy (LHON) can be difficult to distinguish from optic neuritis due to multiple sclerosis (MS). For several decades an association of LHON and MS has been suspected, and within the past 7 years the LHON nucleotide (nt)-3460 and nt-11778 mtDNA mutations have been identified in several patients with MS-like phenotypes. To further study this association, we tested 42 index patients with clinically definite, familial MS for the LHON mtDNA mutations at nt-3460, nt-11778, and nt-14484. No patients had a pathogenic LHON mtDNA mutation; however, two MS patients with unilateral optic neuritis harbored the nt-15257 mtDNA polymorphism that was reported originally as a pathogenic LHON mutation. Several investigators have shown evidence that the nt-15257 mtDNA mutation is not primarily pathogenic. Therefore, we conclude that pathogenic LHON mtDNA mutations are absent or rare in unselected patients with familial, clinically definite MS (95% confidence intervals for each of the negative mutations 0-7.0%).

Adult↗
PubMed1999

Which copper is paramagnetic in the type 2/type 3 cluster of laccase?

Understanding the structure and function of the three copper atoms in the dioxygen reduction site of the blue oxidases such as laccase has been a long standing challenge. In the case of a widely studied derivative, known as type 2-depleted laccase, the removal of one copper from the cluster abolishes the EPR signal of the so-called type 2 copper. However, the present studies of isotopically enriched protein from Polyporus versicolor show that the readily replaceable copper is not active in the low-temperature EPR spectrum of fungal laccase or its difluoride adduct. The same is true for the difluoride adduct of the tree enzyme. Thus, in type 2-depleted laccase the pattern of antiferromagnetic coupling is quite different from that of the native protein or the difluoride adduct.

Copper↗
PubMed1999

Flow cytometric analysis of in vitro proinflammatory cytokine secretion in peripheral blood from multiple sclerosis patients.

The cytokines, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and interleukin-2 (IL-2) are important endogenous proinflammatory proteins and have been linked to disease activity in multiple sclerosis. In this study, we use flow cytometric methodology to compare the secretion of IFN-gamma, IL-2, and TNF-alpha from peripheral blood-derived T cells of multiple sclerosis patients to the secretion in healthy controls. The percentages of IFN-gamma, IL-2, and TNF-alpha secreting cells are not significantly different between multiple sclerosis patients and controls. However, the TNF-alpha secreting CD3 cell percentage is correlated with the IFN-gamma and IL-2 secreting CD3 cell percentages in multiple sclerosis patients. In the controls, only the TNF-alpha secreting CD3 cell percentage is correlated with IFN-gamma. These findings show that correlated secretion of cytokines occurs in multiple sclerosis and suggest that concerted intercytokine interactions may play an important role in the disease.

Adolescent↗
PubMed1999

K+ channels lacking the 'tetramerization' domain: implications for pore structure.

The difficulty of obtaining high-resolution structures of integral membrane proteins has been a frustrating barrier to understanding the membrane-based functions of living cells. The mere handful of such structures stands out in dismal contrast to the cornucopia of water-soluble proteins comprehensible at the atomic level. Nevertheless, crystallographically tractable preparations of aqueous domains of membrane proteins have provided molecular insight into phenomena as varied as chemotaxis, immune cell responses to antigens, viral infectivity and cellular synthesis of ATP. Recently, the first structural glimpse of a neuronal ion channel was reported - the T1, or 'tetramerization,' domain of a Shaker-type voltage-gated K+ channel at 1.6 A resolution. The isolated domain associates into a water-soluble four-fold symmetric homotetramer. This structure prompted the novel, provocative proposal that the T1 domain is an essential component of the ion permeation pathway, forming a previously unsuspected ion-coordinating constriction on the cytoplasmic side of the channel and acting as the receptor for the pore-blocking 'ball and chain' inactivation peptide. It has also been commonly conjectured that the T1 domain is required for tetramerization in the channel maturation process. By studying the detailed properties of Shaker K+ channels in which the T1 domain is deleted, we show all these proposals to be invalid. The structure of the T1 domain expressed in isolation is therefore unlikely to mirror in detail its structure when attached to the ion-conducting channel.

Animals↗
PubMed1999

Immune modulation in autologous bone marrow transplantation: cyclosporine and gamma-interferon trial.

From March 1994 to November 1994, 16 patients with high risk hematological malignancies were entered in a phase I clinical trial, designed to confirm the toxicity of cyclosporine and gamma interferon given to induce autologous graft-versus-host disease (GVHD) after autologous bone marrow transplantation (ABMT). This trial was based on the results in a rodent model, in which cyclosporine given after ABMT induces an autoimmune syndrome (autologous GVHD) identical to allogeneic GVHD. Further, this autologous GVHD is associated with a graft-versus-tumor effect augmented by interferon that upregulates MHC class II expression on normal and tumor cells, the target of the cytolytic T cells in autologous GVHD. In this trial, cyclosporine 1 mg/kg/day was given from the day of bone marrow reinfusion until the completion of the interferon and gamma-interferon. Gamma-interferon at 0.025 mg/m2 every other day was started when the total white cell count was >200 cells/ml for 2 consecutive days and continued for a total of 10 doses after ABMT. The preparative regimens were busulfan and cyclophosphamide, or cyclophosphamide with total body irradiation. All patients received 4HC-purged marrow grafts. Median age was 45 years (range 19-68). The diagnoses included chemo-resistant non-Hodgkin's lymphoma (10), acute lymphoblastic leukemia (two), chemo-resistant Hodgkin's disease (two), acute myeloid leukemia (one), and multiple myeloma (one). Median absolute neutrophil count recovery was 25.5 days (range 19-46 days). Median platelet count recovery was 40.5 days (range 28-279 days). There were nine deaths, two were related to transplant toxicity (infection), while the other seven were due to relapse. Event-free survival with a median of 964 days (range 19-1441 days of follow-up was 44%. In conclusion, treatment with cyclosporine, and gamma-interferon after ABMT was well tolerated and did not impair engraftment. Further studies with a larger number of patients are required to document any beneficial anti-tumor effect of autologous GVHD induction after ABMT.

Adolescent↗
PubMed1999

Cell division tracking and expansion of hematopoietic long-term repopulating cells.

The combined use of rigorous assays for quantitating transplantable stem cell numbers and precise cell labeling and tracking procedures have provided definitive evidence that stem cell self-renewal divisions can occur in vitro in the absence of stromal feeder layers. These findings set the stage for defining conditions that may alter the ability of these cells to maintain their primitive status when mitogenically activated.

Animals↗
PubMed1999

Grammatical morphology and the lexicon in children with specific language impairment.

We examined the use of grammatical morphology by preschool-age English-speaking children with specific language impairment (SLI) as a function of their lexical diversity. Relative to a group of normally developing (ND) preschoolers, these children's use of finite-verb morphology logged behind expectations based on the number of different verbs they used. Noun-related morphology fell below expectations based on overall lexical diversity. Differences between the ND children and children with SLI were also seen for the slope of the increases in finite-verb morphology as a function of lexical diversity, with shallower slopes in the SLI data. The findings of this study add to existing evidence suggesting that a measure of finite grammatical-morphology use has promise as a clinical marker of SLI in English.

Age Factors↗
PubMed1999

Issues in balancing teenage clients' confidentiality and reporting statutory rape among Kansas Title X clinic staff.

Through Federal welfare reform, Congress directed states to aggressively enforce statutory rape laws. Family planning professionals deal with many adolescent clients, and their support for such enforcement or willingness to report is unclear. The authors of this study examined current attitudes and practices of family planning program managers (FPPMs) about statutory rape law enforcement, including current reporting practices. In 1997, all 77 local Kansas Title X FPPMs were surveyed. Structured telephone interviews were conducted with 10 FPPMs to add detail to quantitative responses. Sixty-eight FPPMs responded to the written survey (88%). Of these, 79% supported aggressive enforcement, and 43% thought enforcement would reduce adolescent pregnancy rates. With increased enforcement, 38% believed teenagers would be discouraged from seeking reproductive health care, compared to 41% who believed they would not. Among key informants, all of whom were FPPMs, willingness to report cases was mixed, with those who would report wanting the flexibility to judge on a case-by-case basis. For those not reporting cases, confidentiality concerns overrode beliefs in any positive outcome of enforcement. Kansas Title X FPPMs strongly supported aggressive enforcement, but had mixed beliefs about negative consequences. Among those interviewed, there were also mixed beliefs and practices about reporting. Reporting from FPPMs will be sporadic and arbitrary unless protocols are developed and laws are clarified.

Adolescent↗
PubMed1999

Helical structure and packing orientation of the S2 segment in the Shaker K+ channel.

Six transmembrane segments, S1-S6, cluster around the central pore-forming region in voltage-gated K+ channels. To investigate the structural characteristics of the S2 segment in the Shaker K+ channel, we replaced each residue in S2 singly with tryptophan (or with alanine for the native tryptophan). All but one of the 23 Trp mutants expressed voltage-dependent K+ currents in Xenopus oocytes. The effects of the mutations were classified as being of low or high impact on channel gating properties. The periodicity evident in the effects of these mutations supports an alpha-helical structure for the S2 segment. The high- and low-impact residues cluster onto opposite faces of a helical wheel projection of the S2 segment. The low-impact face is also tolerant of single mutations to asparagine. All results are consistent with the idea that the low-impact face projects toward membrane lipids and that changes in S2 packing occur upon channel opening. We conclude that the S2 segment is a transmembrane alpha helix and that the high-impact face packs against other transmembrane segments in the functional channel.

Amino Acid Sequence↗