Stable carbon and oxygen isotopic compositions of recent charophyte oosporangia and water from Malham Tarn,U.K.: palaeontological implications

Charophyte oosporangia and water samples from a highly calcareous lake were measured for stable carbon and oxygen isotopic composition. The time period over which the oosporangia calcify is short, thus any biochemical relationship between the water and oosporangia"s calcite represents only one time window (late Summer in Malham Tam). This important temporal restraint must also apply to interpretations of all fossil material measured. The ¹⁸O_c of the charophyte oosporangia is deduced to be in equilibrium with the ¹⁸O of the water for a given temperature. The ¹³ C_c of the charophyte oosporangia was approximately 2.5 per mil lower than the ¹³C_DIC in the water we measured. With the release Of CO₂ with phosphoric acid from the charophyte oosporangia, there was no significant difference in the ¹⁸O_c values obtained, regardless of whether or not the carbonate was separated from the organic center, however ¹³C_c values were marginally lower for carbonate plus organic center measurements. Our results indicate that fossil charophyte gyrogonites can be used to elucidate the geochemistry of the ancient water body in which they lived.     

Lactotransferrin is the major estrogen inducible protein of mouse uterine secretions

Antibody to an estrogen inducible mouse uterine protein (Teng, C. T., Walker, M. P., Bhattacharyya, S. N., Klapper, D. G., DiAugustine, R. P., and McLachlan, J. A. (1986) Biochem. J. 240, 413-422) has been used to isolate cDNA to the messenger RNA. Analysis of the deduced primary structure and additional biochemical characterization indicates that the protein is lactotransferrin. An increase in the level of lactotransferrin mRNA of at least 300-fold can be induced in the mouse uterus by estrogen. In contrast, the mRNA is virtually undetectable in rat uterine tissue following estrogen administration. The estrogenic stimulation in mouse uterus contrasts with the known prolactin dependence in mammary gland.     

Estrogen regulation of creatine kinase-B in the rat uterus

Creatine kinase-B (CKB) synthesis is rapidly and specifically induced by estrogen in the uterus of the immature rat. This study indicates that this elevation is due at least in part to increases in the levels of mRNA for CKB. The stimulation of CKB mRNA levels is rapid (a 7- to 10-fold increase is detected 1-3 h after estrogen administration), but transient, as levels return to near control values by 6 h. Analysis of cDNAs to both uterine and brain CKB mRNA indicate that the same sequence is expressed in both tissues despite earlier observations of heterogeneity of the protein isolated from the two tissues. A 1.7-kilobasepair DNA fragment containing the CKB promoter and 5' flanking sequences confers estrogen sensitivity on expression of the bacterial chloramphenicol acetyl transferase gene in HeLa cells on cotransfection with an estrogen-receptor expression vector. However, the CKB promoter sequences lack any motif with convincing similarity to the currently accepted consensus estrogen response element GGTCAnnnTGACC.     

Delineation of sites mediating estrogen regulation of the rat creatine kinase B gene.

We have previously confirmed the estrogen-induced protein of rat uterus to be creatine kinase B (CKB), and demonstrated a 1.7-kilobase pair fragment encompassing the promoter and adjoining 5'-flank to be capable of conferring estrogen responsiveness in HeLa cells. In this study we find an element at -550, aGGTCAgaaCACCCt, with limited similarity to the estrogen response element consensus, to be involved in conferring estrogen responsiveness on the CKB promoter. This element can bind estrogen receptor (ER) and is flanked by two GC boxes, which we find capable of binding bacterially expressed Sp1. Additional responsiveness is found closely associated with the CKB promoter at high levels of cotransfected ER construct. No potential response element was identified in this region, but we find the ER DNA-binding domain to be required.     

Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin, 12-O-tetradecanoylphorbol-13-acetate and 17β-estradiol on estrogen receptor regulation in MCF-7 human breast cancer cells

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) exhibits remarkably potent antiestrogenic activity. To further elucidate the role of estrogen receptor (ER) regulation in this response, we examined the effects of exposure to TCDD in MCF-7 human breast cancer cells on ER mRNA levels by using an RNase protection assay, on ER accumulation by using an ER immunocytochemical essay (ER-ICA), and on ER function by competitive binding assays under conditions of saturating 17β-estradiol (E₂). Comparative studies were conducted with E₂ and 12-O-tetradecanoylphorbol-13-acetate (TPA), as both compounds are known to suppress ER expression. Our results indicate that 1 nM E₂ and 100 nM TPA both suppress ER mRNA levels as early as 4 h after exposure and to 33.6% and 16.5% of control levels, respectively, after 72 h. In contrast, no significant effect on ER mRNA levels was attributed to exposure to 10 nM TCDD. A greater than 50% reduction in positive staining was observed by ER-ICA after 72 h exposure to 1 nM E₂ and to 100 nM TPA, while only an 11% reduction in positive staining was observed with 10 nM TCDD. Specific binding of [³H]E₂ under saturating conditions (10 nM E₂) in whole cells was reduced by 50% in cultures exposed to 100 nM TPA, although no effect on binding was observed with exposure to 10 nM TCDD. In contrast, specific binding using subsaturating 1 nM [³H]E₂ was depressed by 49% in MCF-7 cells exposed to 10 nM TCDD for 72 h. This depression was inhibited by a 1-h treatment with 5 μM α-naphthoflavone, which inhibits TCDD-induced, P450-mediated, E₂ metabolism, and subsequent E₂ depletion. In conclusion, while TPA and E₂ effectively down-regulate ER expression, TCDD, under antiestrogenic conditions, has little if any effect on total ER levels in MCF-7 cells, and thus ER modulation is probably not necessary for the suppression of estrogenic activity in MCF-7 cells by TCDD. © 1996 Wiley-Liss, Inc.     

Anticancer Activities of 7-, and 9-Substituted 3-Deazaguanine Derivatives

Abstract

Interesting differences in anticancer activities of 7- and 9-substituted derivatives of 3-deazaguanine are described.     

Suppression of Ventricular Tachyarrhythmias by Transvenous Intracardiac Pacing after Acute Myocardial Infarction

Transvenous intracardiac pacing was carried out on nine patients with recurrent ventricular tachyarrhythmia after drug treatment had failed. In eight patients recurrent Ventricular ectopic activity was suppressed by pacing at a rate above the sinus rate. With this technique the need for D.C. shock and repeated antiarrhythmic drugs can be avoided.     

Evidence for Ubiquitin-Regulated Nuclear and Subnuclear Trafficking among Paramyxovirinae Matrix Proteins

The paramyxovirus matrix (M) protein is a molecular scaffold required for viral morphogenesis and budding at the plasma membrane. Transient nuclear residence of some M proteins hints at non-structural roles. However, little is known regarding the mechanisms that regulate the nuclear sojourn. Previously, we found that the nuclear-cytoplasmic trafficking of Nipah virus M (NiV-M) is a prerequisite for budding, and is regulated by a bipartite nuclear localization signal (NLS_bp), a leucine-rich nuclear export signal (NES), and monoubiquitination of the K258 residue within the NLSbp itself (NLS_bp-lysine). To define whether the sequence determinants of nuclear trafficking identified in NiV-M are common among other Paramyxovirinae M proteins, we generated the homologous NES and NLS_bp-lysine mutations in M proteins from the five major Paramyxovirinae genera. Using quantitative 3D confocal microscopy, we determined that the NES and NLS_bp-lysine are required for the efficient nuclear export of the M proteins of Nipah virus, Hendra virus, Sendai virus, and Mumps virus. Pharmacological depletion of free ubiquitin or mutation of the conserved NLS_bp-lysine to an arginine, which inhibits M ubiquitination, also results in nuclear and nucleolar retention of these M proteins. Recombinant Sendai virus (rSeV-eGFP) bearing the NES or NLS_bp-lysine M mutants rescued at similar efficiencies to wild type. However, foci of cells expressing the M mutants displayed marked fusogenicity in contrast to wild type, and infection did not spread. Recombinant Mumps virus (rMuV-eGFP) bearing the homologous mutations showed similar defects in viral morphogenesis. Finally, shotgun proteomics experiments indicated that the interactomes of Paramyxovirinae M proteins are significantly enriched for components of the nuclear pore complex, nuclear transport receptors, and nucleolar proteins. We then synthesize our functional and proteomics data to propose a working model for the ubiquitin-regulated nuclear-cytoplasmic trafficking of cognate paramyxovirus M proteins that show a consistent nuclear trafficking phenotype.     

Formation of laminate travertines at bagno vignone,Italy

Travertine at Bagno Vignoni, Tuscany, is deposited from thermal CO₂‐rich water up‐welling from an artificial pool, Bagno Grande, which contains a thick benthic mat of cyanobacteria. The travertine mound, over 40 m in height, consists of calcite feather crystals and micrite. Over much of the mound, but particularly in the lower part, the travertine is finely banded, with pale bands of feather crystals alternating with dark bands containing micrite. Deposition rate measurements showed that the bands were formed daily, with the slightly narrower and darker bands formed at night. These nocturnal bands also contained wind‐borne dust. It is postulated that the dust caused heterogeneous nucleation of micrite and reduced the development of feather crystals. The deposition rate of travertine ranged from 127 to 348 μm/day and was highest at the top of the mound. Water analysis conducted over 27 h demonstrated a pronounced diurnal variation in the dissolved carbon dioxide of Bagno Grande, consistent with the photosynthetic activity of the benthic cyanobacteria. This variation could be traced to the base of the travertine mound but was not alone responsible for the higher deposition rate during the day. Water temperature on the mound also increased during the day and would have aided CO₂ evasion. Photosynthetic activity on the mound itself appeared to have far less influence on the carbon dioxide flux than Bagno Grande. The upper mound supported unlithified mats of the cyanobacteria Lyngbya (Phormidium) laminosum and Spirulina labyrinthiformis. These mats sometimes possessed fine travertine laminae resulting from cyanobacterium phototaxis and may have been responsible for some lamination in the upper mound. The CO₂ flux measurements indicated that overall, atmospheric evasion of carbon dioxide was more significant to travertine deposition than photosynthetic activity.