Human interleukin 4 receptor confers biological responsiveness and defines a novel receptor superfamily.

IL-4, a pleiotropic cytokine produced by T lymphocytes, plays an important role in immune responsiveness by regulating proliferation and differentiation of a variety of lymphoid and myeloid cells via binding to high affinity receptors. In this report we describe the isolation and functional expression of a human IL-4-R cDNA. When transfected into COS-7 cells, the cDNA encodes a 140-kD cell-surface protein. After transfection into a murine T cell line, the cDNA encodes a protein that binds human IL-4 with high affinity and can confer responsiveness to human IL-4. The predicted extracellular domain of the IL-4-R exhibits significant amino acid sequence homology with the beta subunit of the IL-2-R (p75), and the receptors for IL-6, erythropoietin, and prolactin. These receptors comprise a novel superfamily with extracellular domains characterized by four conserved cysteine residues and a double tryptophan-serine (WSXWS) motif located proximal to the transmembrane region.

Molecular cloning of the interleukin-1 beta converting enzyme.

Interleukin-1 beta (IL-1 beta) mediates a wide range of immune and inflammatory responses. The active cytokine is generated by proteolytic cleavage of an inactive precursor. A complementary DNA encoding a protease that carries out this cleavage has been cloned. Recombinant expression in COS-7 cells enabled the cells to process precursor IL-1 beta to the mature form. Sequence analysis indicated that the enzyme itself may undergo proteolytic processing. The gene encoding the protease was mapped to chromosomal band 11q23, a site frequently involved in rearrangement in human cancers.

The IL-6 signal transducer, gp130: an oncostatin M receptor and affinity converter for the LIF receptor.

Leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) are multifunctional cytokines with many similar activities. LIF is structurally and functionally related to another cytokine, Oncostatin M (OSM), that binds to the high-affinity LIF receptor but not to the low-affinity LIF receptor. A complementary DNA was isolated that encodes the high-affinity converting subunit of the LIF receptor. The converter conferred high-affinity binding of both LIF and OSM when expressed with the low-affinity LIF receptor and is identical to the signal transducing subunit of the IL-6 receptor, gp130. The gp130 subunit alone confers low-affinity binding of OSM when expressed in COS-7 cells. This receptor system resembles the high-affinity receptors for granulocyte-macrophage colony-stimulating factor, IL-3, and IL-5, which share a common subunit.

Maternal exposure to water disinfection by-products during gestation and risk of hypospadias.

BACKGROUND: The use of chlorine for water disinfection results in the formation of numerous contaminants called disinfection by-products (DBPs), which may be associated with birth defects, including urinary tract defects. METHODS: We used Arkansas birth records (1998-2002) to conduct a population-based case-control study investigating the relationship between hypospadias and two classes of DBPs, trihalomethanes (THM) and haloacetic acids (HAA). We utilised monitoring data, spline regression and geographical information systems (GIS) to link daily concentrations of these DBPs from 263 water utilities to 320 cases and 614 controls. We calculated ORs for hypospadias and exposure to DBPs between 6 and 16 weeks' gestation, and conducted subset analyses for exposure from ingestion, and metrics incorporating consumption, showering and bathing. RESULTS: We found no increase in risk when women in the highest tertiles of exposure were compared to those in the lowest for any DBP. When ingestion alone was used to assess exposure among a subset of 40 cases and 243 controls, the intermediate tertiles of exposure to total THM and the five most common HAA had ORs of 2.11 (95% CI 0.89 to 5.00) and 2.45 (95% CI 1.06 to 5.67), respectively, compared to women with no exposure. When exposure to total THM from consumption, showering and bathing exposures was evaluated, we found an OR of 1.96 (95% CI 0.65 to 6.42) for the highest tertile of exposure and weak evidence of a dose-response relationship. CONCLUSIONS: Our results provide little evidence for a positive relationship between DBP exposure during gestation and an increased risk of hypospadias but emphasise the necessity of including individual-level data when assessing exposure to DBPs.

Distinct regions of the human granulocyte-colony-stimulating factor receptor cytoplasmic domain are required for proliferation and gene induction.

Using two different cell systems, we show that the cytoplasmic domain of the granulocyte-colony-stimulating factor receptor (G-CSFR) may be composed of at least two functional regions. The first, within the membrane-proximal 57 amino acids, is absolutely required to deliver a proliferative signal. This region contains two sequence motifs conserved between members of the hematopoietin receptor family. The second functional region resides between amino acids 57 and 96. This region is required for the induction of acute-phase plasma protein gene expression when the G-CSFR is transfected into human hepatoma cell lines. The G-CSFR-transfected hepatoma cells respond to G-CSF by increasing the production of the same set of plasma proteins as stimulated by interleukin-6, suggesting that the two cytokines share a common signal transduction pathway.

A spectrum project: preterm birth and small-for-gestational age among infants with birth defects.

OBJECTIVE: The aim of this study is to investigate the association between birth defects (BDs), prematurity and small-for-gestational age (SGA) in a population-based sample. STUDY DESIGN: Participants were singleton live births enrolled in the National Birth Defects Prevention Study, including 18 737 case infants with one or more BD and 7999 controls. Logistic regression models to evaluate associations between BDs, prematurity and fetal growth were computed while adjusting for covariates. RESULT: Cases were significantly more likely to be born prematurely than controls, particularly at 24 to 28 weeks of gestation. The highest odds ratios for preterm birth were found for intestinal atresia, anencephaly, gastroschisis and esophageal atresia. Infants with BDs were also significantly more likely to be SGA than controls (17.2 and 7.8%). CONCLUSION: Infants with BDs are more likely than controls to be born prematurely and SGA. Findings from this study present additional evidence demonstrating a complex interaction between the development of BDs, prematurity and intrauterine growth.

On the accuracy and reliability of predictions by control-system theory.

In three experiments we used control-system theory (CST) to predict the results of tracking tasks on which people held a handle to keep a cursor even with a target on a computer screen. 10 people completed a total of 104 replications of the task. In each experiment, there were two conditions: in one, only the handle affected the position of the cursor; in the other, a random disturbance also affected the cursor. From a person's performance during Condition 1, we derived constants used in the CST model to predict the results of Condition 2. In two experiments, predictions occurred a few minutes before Condition 2; in one experiment, the delay was 1 yr. During a 1-min. experimental run, the positions of handle and cursor, produced by the person, were each sampled 1800 times, once every 1/30 sec. During a modeling run, the model predicted the positions of the handle and target for each of the 1800 intervals sampled in the experimental run. In 104 replications, the mean correlation between predicted and actual positions of the handle was .996; SD = .002.

A national survey of current admission practices in selected allied health educational programs.

This study investigated the admission practices of selected allied health programs, specifically considering the priorities placed on cognitive and noncognitive factors. The extents to which diversity and a student's desire to work in underserved areas are considered in the selection process were also examined. Of 206 questionnaires mailed to accredited baccalaureate occupational therapy, physical therapy, health information management, and respiratory therapy programs, 144 were returned. The results indicate that allied health programs use combinations of cognitive and noncognitive factors in selecting students for admission. A higher priority is placed on overall grade-point average (GPA) and GPA in foundation courses, whereas lower priorities are placed on the need for diversity and a student's desire to work in underserved areas. The authors discuss the implications of the findings and urge the rethinking of the traditional selection method, which places applicants from ethnic/racial minority backgrounds at a disadvantage.

Determination of the minimum polypeptide lengths of the functionally active sites of human interleukins 1 alpha and 1 beta.

Interleukin 1 (IL-1) is a two-member family of proteins (IL-1 alpha and IL-1 beta) that mediates a diverse series of immune and inflammatory responses. These two proteins have only 26% amino acid homology yet bind to the same receptor. It is of importance to define the active sites of these molecules in order to understand their receptor interactions and the mechanisms involved in their multiple biological functions. We report here the localization of the biologically active portions within the initial polypeptide translation products. An in vitro transcription and translation system was used to generate specific fragments of each of the IL-1 molecules, which then were assayed for receptor binding capability and biological activity. Using this system, we have demonstrated that core sequences of amino acids for IL-1 beta (numbers 120-266) and 140 amino acids for IL-1 alpha (numbers 128-267) must be left intact to retain full biological activity and further that the biological activities of the IL-1 polypeptides parallel their receptor binding capabilities.

Black adolescents' attitudes toward and use of alcohol and other drugs.

Substance use and misuse has become a major problem in recent years. Although there has been research conducted concerning the use of substances by adolescents, little attention has been focused on the Black adolescent. The purpose of this research was to identify the attitudes of Black adolescents toward substances, their level of use, and the extent of their participation in alternative activities. Results indicated that there was a high percentage of students who reported negative attitudes toward substances, that actual use levels were extremely low, and that students spend a considerable amount of time involved in alternative activities.

Influence of subunit combinations on signaling by receptors for oncostatin M, leukemia inhibitory factor, and interleukin-6.

Oncostatin M (OSM), leukemia inhibitory factor (LIF), and interleukin-6 (IL-6) induce expression of a similar set of acute phase plasma protein genes in hepatic cells. The redundant action of these cytokines has been ascribed to the involvement of the common signal-transducing receptor subunit, gp130, in combination with cytokine-specific, ligand-binding subunits. To define the specificity of the signal transduction by the LIF/OSM receptor (a heterodimer of gp130 and LIF receptor (LIFR)) and the OSM-specific receptor (a heterodimer of gp130 and OSM receptor (OSMR)), we reconstituted the receptor function by transfection into receptor-negative Hep3B hepatoma cells. Both receptors activate DNA binding activity of STAT1, -3, and -5B and induce gene transcription through IL-6-responsive elements. The signaling-competent cytoplasmic domain regions of OSMR and LIFR were defined by the analysis of progressive carboxyl-terminal deletion constructs. The 36 residue carboxyl-terminal region containing the distal box 3 sequence motif of OSMR is required for signal transduction by the OSM-specific receptor. In contrast, signaling by LIFR did not display the same requirement for receptor domains and was not strictly dependent on the box 3 elements. The signaling by endogenous LIF and OSM receptors differed from that by IL-6R by the prominent activation of STAT5 as shown in the mouse hepatoma cell line, Hepa-1. The data suggest that the signaling specificity of the receptors for the three cytokines is determined by the composition of the cytoplasmic domains associated in the signal-competent receptor complex and that the signaling is not identical among these cytokine receptors.

Oncostatin M-specific receptor expression and function in regulating cell proliferation of normal and malignant mammary epithelial cells.

Oncostatin M (OSM) is a cytokine produced by activated T lymphocytes and macrophages. OSM is structurally and functionally related to leukaemia inhibitory factor (LIF), another cytokine in the interleukin 6 (IL-6) family. The biological activities of OSM are mediated through two types of receptor complexes, the LIF/OSM shared receptor (type I) and OSM-specific receptor (OSM-R, type II), which is composed of gp130 as a binding subunit and a newly identified affinity conversion subunit, OSM-R beta. Previous research conducted in the authors' laboratory has shown that OSM inhibits the growth of several breast cancer cell lines. To investigate whether OSM has a similar effect in primary normal human mammary epithelial (HME) cells, the activity of OSM in HME cells derived from four donors was examined. OSM produced a dose-dependent inhibition of DNA synethesis in these cells. In order to determine the receptor subtypes mediating OSM activity in HME and breast cancer cells, flow cytometry analysis using anti-gp130mAb and anti-OSM-R beta mAb was performed. In these studies, the authors were able to examine expressions of gp130 and OSM-R beta. In addition, quantitative RT-PCR assays were conducted to measure expressions of the mRNAs of the subunits for type I and type II OSM receptor. The results show that HME cells and most breast cancer cell lines express both the type I and the type II OSM receptors. However, type II, OSM-specific receptors are expressed at a higher levels than type I, OSM/LIF shared receptors. Accordingly, we compared the growth regulatory activities of OSM with LIF in HME cells and in breast cancer cells. In contrast to the inhibitory activity of OSM, LIF stimulated the growth of breast cancer cells, whereas it had no effect on normal mammary epithelial cell growth. Together, these data suggest that OSM plays an inhibitory role in normal and malignant mammary epithelial cell growth in vitro. OSM activity is mediated by the OSM-specific receptor (type II), not by the OSM/LIF shared receptor.

Purification and initial characterization of a ferret Pneumocystis carinii surface antigen.

The purification and initial characterization of a ferret Pneumocystis carinii surface glycoprotein is described. Previous studies have demonstrated that passive administration of monoclonal antibody recognizing this glycoprotein reduces the severity of P. carinii pneumonitis in animal models of infection. This acidic glycoprotein (approximate isoelectric point, 5.0-5.7) contains both mannose (and/or glucose) and N-acetyl-glucosamine residues. The cross-reactive surface antigen on P. carinii of human origin is also a mannose- (and/or glucose-) containing glycoprotein. Initial biochemical characterization of these surface molecules should aid in understanding the immunobiology of this organism and in developing reliable diagnostic assays for P. carinii pneumonitis.

Dual oncostatin M (OSM) receptors. Cloning and characterization of an alternative signaling subunit conferring OSM-specific receptor activation.

Oncostatin M (OSM) is a cytokine whose structural and functional features are similar to other members of the interleukin (IL)-6 family of cytokines (IL-6, IL-11, leukemia inhibitory factor (LIF), granulocyte colonystimulating factor, ciliary neurotrophic factor, and cardiotrophin-1), many of which utilize gp130 as a common receptor subunit. A biologically active OSM receptor has been previously described that consists of a heterodimer of leukemia inhibitory factor receptor (LIFR) and gp130. This LIFR.gp130 complex is also a functional receptor for LIF. We have cloned and characterized an alternative subunit (OSMRbeta) for an OSM receptor complex (a heterodimer of gp130 and OSMRbeta) that is activated by OSM but not by LIF. The signaling capability of specific receptor subunit combinations was analyzed by independent assays measuring cell proliferation or induction of acute phase protein synthesis. Our results demonstrate that both LIF and OSM cause tyrosine phosphorylation and activation of the gp130.LIFR combination, but the gp130.OSMRbeta complex is activated by OSM only. OSM-induced cellular responses, initiated through low affinity binding to gp130, are mediated by two heterodimeric receptor complexes that utilize alternative signal transducing subunits that confer different cytokine specificities to the receptor complex.

Percutaneous ultrasonic lithotripsy in the community hospital.

A total of 38 patients underwent attempted percutaneous extraction of upper tract calculi with the Wolf nephroscope. A 1-day interval between radiological insertion of the guide wire and extraction of the stone with the patient under general anesthesia was used. Of the stones 87 per cent were removed successfully. Average operating and hospitalization times were 62 minutes and 9.9 days, respectively. Reduced morbidity and a shorter interval to return to work make this the procedure of choice in selected cases.

Defense of the radical perineal prostatectomy.

There is sharp disagreement as to what constitutes the proper surgical approach to localized carcinoma of the prostate. We have performed 31 radical perineal prostatectomies in a six-year period with no mortality and minimal morbidity. Thirteen of these patients were understaged preoperatively and had extraprostatic cancer; however, only one has died from his tumor. One patient is incontinent but none has troublesome local symptoms. These patients required an average of 15 postoperative days, none required more than two units of blood, and careful preoperative consultation has minimized the psychologic stress of impotence. These data contrast sharply with the published morbidity and mortality statistics associated with a preliminary staging lymphadenectomy and a definitive radical retropubic prostatectomy. Also, we are convinced that our patients with stage C cancer have been done a real service by removing the prostate gland even though cancer remains in the stumps of the seminal vesicles. Unless the advocates of the staged procedure can demonstrate an improvement in the patients' survival data, we believe the radical perineal prostatectomy remains the procedure of choice for the cure of localized prostatic cancer and we would advocate this operation as an acceptable palliative approach to selected patients with stage C lesions.

Reconstitution of the response to leukemia inhibitory factor, oncostatin M, and ciliary neurotrophic factor in hepatoma cells.

Ciliary neurotrophic factor (CNTF) has been described as a neuro-active cytokine that shares functional similarities with the leukemia inhibitory factor (LIF). We demonstrate here that, like LIF, CNTF stimulates expression of acute phase plasma proteins in rat H-35 hepatoma cells. Transfection of the LIF receptor into Hep3B hepatoma cells reconstituted LIF and oncostatin M regulation of acute phase plasma protein genes. Co-expression of the LIF receptor and the CNTF receptor, but not expression of either subunit alone, generated CNTF responsiveness in Hep3B cells, suggesting cooperativity of these receptor subunits. Evidence is presented for direct interaction of the LIF receptor with the intracellular signal transduction machinery.