High-frequency1H NMR studies of the effects of growth factors and phorbol esters on normal syrian hamster diploid fibroblast cells

The nuclear magnetic resonance (NMR) parameters, spin-lattice (T₁), and spin-spin (T₂) relaxation time, are usually longer for neoplastic cells than for normal cells of the same cell type. This has generally been true at low NMR frequencies (≤100 MHz) when comparisons have been made between normal and neoplastic cells that have both spent a short time in culture. We have previously demonstrated that although the T₁ values of paired normal and neoplastic Syrian hamster (SH) fibroblastic cells in culture are not significantly different when measured at 300 MHz, the 300 MHz T₂ values for the neoplastic cells are smaller than those of the normal cells. (Xin et al. (1986),Cell Biophysics 8, 213.) Since treatment of normal diploid cells with polypeptide growth factors or tumor promoters frequently results in reversible expression of neoplasia-associated phenotypes, T₁ and T₂ were obtained at 300 MHz for treated and untreated SH cells to see if these compounds could also produce smaller 300 MHz T₂ values. Secondary culture SH fetal fibroblast cells were treated with epidermal growth factor (EGF), fibroblast growth factor (FGF), phorbol-12,13-didecanoate (PDD) and 4-α-phorbol-12,13-didecanoate (4αPDD). Treatment with either growth factor resulted in smaller T₂ values, but a statistically significant decrease was not observed for PDD or 4αPDD. The observed reductions in T₂ values were correlated with the morphological and growth-stimulatory effects of these compounds on the cells.     

Relationship between inspiratory effort and breathlessness in pregnancy.

Using open-magnitude scaling, we compared the relationships between breathlessness, inspiratory esophageal pressure swing (delta Pes), and ventilation in pregnancy and postpartum. Thirteen healthy women performed progressive cycle exercise tests at 33 +/- 2 wk gestation and 12 +/- 3 wk postpartum. Pulmonary function and maximal transdiaphragmatic pressure did not change. Minute ventilation (VE) was greater in the third trimester. This increase was entirely due to the increase in tidal volume (VT; 0.74 +/- 0.18 vs. 0.54 +/- 0.18 liters at rest, P less than 0.01; 1.56 +/- 0.3 vs. 1.24 +/- 0.24 liters at 48 W, P less than 0.001). delta Pes (15.3 +/- 3.0 vs. 11.9 +/- 3.5 cmH2O at 48 W, P less than 0.01) and breathlessness (1.8 +/- 1.4 vs. 1.0 +/- 0.9 at 48 W, P less than 0.05) were greater in the third trimester. However, the relationships between VT and delta Pes and between delta Pes and breathlessness were identical in the two conditions. The VT-tidal abdominal volume (Vab) and Vab-tidal gastric pressure swing (delta Pga) relationships were similar in the two conditions. In conclusion, the relationship between delta Pes and breathlessness is the same in the third trimester and postpartum. The increased VE is responsible for the breathlessness in the third trimester. Despite progressive abdominal distension by the gravid uterus, the VT-Vab and Vab-delta Pga relationships were the same in the two conditions.     

Human non-lineage antigen,CD46 (HuLy-m5): purification and partial sequencing demonstrates structural homology with complement-regulating glycoproteins

CD46, until recently known as HuLy-m5, is a non-lineage restricted surface antigen ubiquitously expressed by almost all human cells except erythrocytes. The CD46 antigen is identified by the E4.3 monoclonal antibody (mAb) and exists at the surface of human peripheral blood lymphocytes (PBLs) as two acidic, non-disulfide bonded chains, and , ofM _r 66 000 and 56 000. Receptor density analysis showed that CD46 was of moderately low abundance on PBLs with 7.5×10³ molecules present on each cell. The two chains of CD46 were purified (144 000-fold) by immunoaffinity-chromatography with E4.3 mAb from the plasma membranes of a human spleen infiltrated with chronic myelogenous leukemia cells. Amino acid sequence analysis of the NH₂-terminal of both and chains yielded the same sequence; XEEPPQ/TFEAMELIGKPKPYYEIGE. Peptide mapping studies confirmed that both CD46 chains were closely related, except for one peptide fragment. This amino acid sequence is identical to that of the NH₂-terminal of the recently cloned membrane co-factor protein (MCP), a membrane protein that binds the C3b and C4b fragments of complement and acts as a co-factor for I protein-mediated decay of the complement convertases. CD46 shares a cross-reactive epitope with some primate retroviruses, and this may indicate that some retroviruses mimic the mechanisms used by autologous human cells to evade complement-mediated immune clearance. Offprint requests to: I. F. C. McKenzie.     

Mutation analysis of the cystic fibrosis transmembrane regulator gene in native American populations of the southwest

We report DNA and clinical analyses of cystic fibrosis (CF) in two previously unstudied, genetically isolated populations: Pueblo and Navajo Native Americans. Direct mutation analysis of six mutations of the CFTR gene--namely, delta F508, G542X, G551D, R553X, N1303K, and W1282X--was performed on PCR-amplified genomic DNA extracted from blood samples. Haplotype analyses with marker/enzyme pairs XV2c/TaqI and KM19/PstI were performed as well. Of the 12 affected individuals studied, no delta F508 mutation was detected; only one G542X mutation was found. None of the other mutations was detected. All affected individuals have either an AA, AC, or CC haplotype, except for the one carrying the G542X mutation, who has the haplotype AB. Clinically, six of the affected individuals examined exhibit growth deficiency, and five (all from the Zuni Pueblo) have a severe CF phenotype. Four of the six Zunis with CF are also microcephalic, a finding not previously noted in CF patients. Our DNA data have serious implications for risk assessment of CF carrier status for these people.     

Barley anther culture: Pretreatment on mannitol stimulates production of microspore-derived embryos

A period of four days preincubation at 25 °C on a medium containing mannitol was found to be superior to those pretreatments requiring incubation at 4 °C. In addition, the yield of green plants was improved by orienting anthers flat on the medium during mannitol preincubation, and reducing the number of anthers cultured per dish.     

Two classes of continuous cell lines established from Syrian hamster 9 day gestation embryos: Preneoplastic cells and progenitor cells

Summary Primary cultures of 9-d-gestation Syrian hamster embryo (E9) cells are distinct from primary cultures of later gestational age in terms of their growth and differentiation. First, primary E9 cell cultures express multiple mesenchymal differentiation lineages (e.g., adipocyte, myoblast) only rarely seen in cultures of 13-d-gestation fetal (F13) cells. Second, although most primary E9 cultures have a limited in vitro proliferative life span and exhibit cellular senescence similar to primary cultures of F13 cells, E9 cultures seem to have higher frequency of escape from senescence and conversion to continuous cell lines compared to F13 cells. Moreover, this frequency can be further increased 4- to 5-fold by continuous exposure of the E9 cells to tumor promoters or epidermal growth factor. Eleven continuous cell lines have been isolated from unreated, promoter-treated, or epidermal growth factor-treated primary E9 cultures. Seven of these are neoplastic or preneoplastic. However, the remaining four do not show any evidence of being in neoplastic progression and three of these continue to express the same differentiated phenotype observed in ther parental primary cell cultures. These studies were supported in part by grants from the National Institutes of Health (AG 01998), Bethesda, MD, and the U.S. Department of Energy (DE-A-C02-76-EVO-3280), Washington, DC.     

The microbial plankton of Lake Fryxell,southern Victoria Land,Antarctica during the summers of 1992 and 1994

 Samples collected from Lake Fryxell, southern Victoria Land, Antarctica in January 1992 and 1994 were analysed for the abundance of bacterioplankton and the diversity and abundance of protistan plankton. At the times of sampling, 14 ciliate species and 10 species of autotrophic flagellate were recorded. The samples contained two species of rotifer (Philodina spp.), which formed the first record of planktonic metazoans in the Dry Valley lakes of this region of Antarctica. Bacterial concentrations ranged between 1.0 and 3.8×10⁸ l^-1 in the upper oxic waters increasing to 20×10⁸ l^-1 in the anoxic waters. Heterotrophic flagellates decreased in abundance down the oxygenated water column, disappearing completely at 9 m, and ranged between 0.28 and 7.39×10⁵ l^-1 in abundance. Autotrophic flagellates were much more abundant exhibiting a number of distinct peaks down the water column (1.89–25.3×10⁸ l^-1). The ciliated protozoa were very abundant (up to 7720 l^-1) in relation to flagellate and bacterial numbers, typical of oligotrophic lakes world-wide. The distribution of the protistan plankton showed marked zonation, probably in response to the differing salinity and temperature gradients in the water column. Possible trophic interactions are discussed and comparisons with other continental Antarctic lakes made. Received: 29 November 1995/Accepted: 18 February 1996