Cytotoxic T-cell-resistant variants arise at early times after infection in C57BL/6 but not in SCID mice infected with a neurotropic coronavirus.

Under certain conditions, C57BL/6 mice persistently infected with mouse hepatitis virus strain JHM (MHV-JHM) develop clinical disease and histological evidence of demyelination several weeks after inoculation with virus. In a previous report, we showed that mutations in the RNA encoding an immunodominant CD8 T-cell epitope within the surface glycoprotein (epitope S-510-518) were present in all persistently infected animals and that these mutations abrogated recognition by virus-specific cytotoxic T cells (CTLs) in direct ex vivo cytotoxicity assays. To obtain further evidence that these mutations were necessary for the development of clinical disease, the temporal course of their appearance was determined. Mutations in the epitope were identified by 10 to 12 days after inoculation, and in some mice, virus containing mutated epitope was the dominant species detected by 15 days. In addition, most mice that remain asymptomatic at 80 days after inoculation, a time after which clinical disease almost never develops, were infected with only wild-type virus. Finally, analysis of virus isolated from mice with severe combined immunodeficiency (SCID) revealed the presence only of wild-type epitope S-510-518. These results, by showing that mutations are not selected in SCID mice and occur at early times after inoculation in C57BL/6 mice, support the view that they result from immune pressure and contribute to virus persistence and demyelination in mice infected persistently with MHV-JHM.     

Differential Regulation of Phenylethanolamine N-Methyltransferase Expression in Two Distinct Subpopulations of Bovine Chromaffin Cells

Abstract: Chromaffin cells were isolated from bovine adrenal glands and fractionated into two distinct subpopulations by density gradient centrifugation on Percoll. Cells in the more dense fraction stored epinephrine (E) as their predominant catecholamine (81% of total catecholamines), contained high levels of phenylethanolamine N-methyltransferase (PNMT) activity, and exhibited intense PNMT immunoreactivity. This population of chromaffin cells was termed the E-rich cell population. Cells in the less dense fraction, the norepinephrine (NE)-rich cell population, stored predominantly NE (75% of total catecholamines). Although the NE-rich cells had only 3% as much PNMT activity as did the E-rich cells, 20% of the NE-rich cells were PNMT immunoreactive. This suggested that the PNMT-positive cells in the NE-rich cell cultures contained less PNMT per cell than did E-rich cells and may not be typical adrenergic cells. The regulation of PNMT mRNA levels and PNMT activity in primary cultures of E-rich and NE-rich cells was compared. At the time the cells were isolated, PNMT mRNA levels in NE-rich cells were ～20% of those in E-rich cells; within 48 h in culture, PNMT mRNA in both populations declined to almost undetectable levels. Treatment with dexamethasone increased PNMT mRNA levels and PNMT activity in both populations. In E-rich cells, dexamethasone restored PNMT mRNA to the level seen in freshly isolated cells and increased PNMT activity twofold. In NE-rich cells, dexamethasone increased PNMT mRNA to levels twice those found in freshly isolated cells and increased PNMT activity sixfold. Cycloheximide blocked the effects of dexamethasone on PNMT mRNA expression in NE-rich cells but had little effect in E-rich cells. Angiotensin II, forskolin, and phorbol 12,13-dibutyrate elicited large increases in PNMT mRNA levels in E-rich cells but had no effect in NE-rich cells. Our data suggest that PNMT expression is regulated differently in the two chromaffin cell subpopulations.     

Yield and water use of wheat (Triticum aestivum) in a Mediterranean environment: Cultivar differences and sowing density effects

Yield of eight wheat cultivars was evaluated under rainfed and irrigated conditions in a Mediterranean environment. Variation in grain yield resulted from variation in both aboveground biomass production and in harvest index. Under rainfed compared to irrigated conditions, grain yield, biomass and days to heading were decreased, whereas harvest index was increased. Grain yield of the different cultivars under rainfed conditions correlated with that under irrigated conditions in one of the two years. Among cultivars, harvest index under rainfed and irrigated conditions were correlated in both years.Water was used more efficiently for biomass production, and equally efficiently for grain production, under irrigated compared to rainfed conditions. Under rainfed conditions, crop water use efficiency was higher for cultivars developed for rainfed environments than for those developed for high-rainfall or irrigated environments. Cultivars with low-rainfall target environments had the lowest evapotranspiration under rainfed conditions. Under rainfed conditions, differences between the cultivar groups in crop water use efficiency corresponded with trends in water use efficiency of individual plants and with the ratio of photosynthesis to transpiration, measured on plants grown in a growth room.Early in the season, water was used more efficiently for biomass production at high sowing densities than at low sowing densities. Through faster biomass production and ground cover a smaller proportion of the evapotranspired water was lost in soil evaporation and a larger proportion was transpired. However, the net effect was a greater water use in the early phases of growth and consequently a lower water availability later in the season, leading to similar yields regardless of sowing density.     

Sulfte-reduction process in sediments of Lake Kinneret,Israel

Monomictic Lake Kinneret is stratified during summer and autumn, resulting in a hypolimnion rich in H₂S (3–7 mg 1^–1). In winter and spring every year a bloom of dinoflagallate Peridinium gatunense produces an average biomass of 150000 ton wet weight. Part of this biomass sinks to the hypolimnion and sediments where it is decomposed and mineralized, with some of the mineralization due to the activity of sulfate-reducing bacteria (SRB). The sulfate-reduction potential of the upper sediment layer at the deepest part of the lake (42 m) was measured. The activity of the enzyme arylsulfatase was also monitored. Rates of sulfate-reduction ranged from a minimum of 12 nmoles SO_f4 ^p2–-reduced cm^–3 day^–1 in December before lake overturn to a maximum of 1673 nmoles SO_f4 ^p2– reduced cm^–3 day^–1 in July during stratification. These rates are considerably higher than those recorded from other freshwater lakes in the world and are probably limited more by the availability of organic matter than by sulfate concentrations.     

24-homologated 1,25-dihydroxyvitamin D3 compounds: separation of calcium and cell differentiation activities

A series of 24-homologated 1,25-dihydroxyvitamin D3 compounds have been chemically synthesized and studied with regard to their activity in inducing differentiation of human promyelocyte HL-60 cells to monocytes and in calcium mobilizing activity in vitamin D deficient rats. Homologation of 1,25-dihydroxyvitamin D3 or its delta 22 analogue by one or two carbons increases by 10-fold and three-carbon homologation reduces by half the activity in causing differentiation of HL-60. On the other hand, homologation causes a substantial decrease in in vivo calcium mobilization activity. The addition of each carbon at the 24-position decreases binding to the HL-60 receptor or rat intestinal receptor by 5-10-fold so that binding affinity of the trihomo compound for the receptors is 130 times less that of 1,25-dihydroxyvitamin D3. Thus, binding affinity for the receptor cannot account for the preferential activity of the 24-homologated compounds in inducing cell differentiation.     

Temporal and spatial root development of cauliflower (Brassica oleracea L. var. botrytis L.)

Row crops are often inefficient in utilizing soil resources. One reason for this appears to be inefficient rooting of the available soil volume. Five experiments were performed to study the temporal and spatial root development of cauliflower (cv. Plana). The crop was grown with 60 cm between rows, and root development was followed in minirhizotrons placed under the crop rows, 15 cm, and 30 cm from the crop rows. Soil was sampled and analyzed for nitrate content at the final harvest and once during growth. In two of the experiments N fertilizer rate was varied and in two of the other experiments two cultivars were compared (cv. Plana and Siria).The rooting depth of cauliflower was found to be linearly related to temperature sum, with a growth rate of 1.02 mm day^-1 °C^-1. Depending on duration of growth this leads to rooting depths at harvest of 85–115 cm. Soil analysis showed that the cauliflower was able to utilize soil nitrogen down to at least 100 cm.With Plana differences in root growth between row and interrow soil were only observed during early growth, but with Siria this difference was maintained until harvest. However, at harvest both cultivars had depleted row and interrow soil nitrate equally efficient. Nitrogen fertilizer did not affect overall root development significantly.The branching frequency of actively branching roots was increased in all soil layers from about 6 to 10 branches cm^-1 by increasing N fertilizer additions from 130 to 290 kg N ha^-1. Increasing N supply increased the number of actively branching roots in the topsoil and reduced it in the subsoil.The average growth rate of the roots was always highest in the newly rooted soil layers, but fell during time. At 74 days after planting very few roots were growing in the upper 60 cm of the soil whereas 70% of the root tips observed in the 80–100 cm soil layer were actively growing. Within each soil layer there was a large variation in growth rate of individual root tips.     

Photocrosslinking of 4-thio uracil-containing RNAs supports a side-by-side arrangement of domains 5 and 6 of a group II intron

Previous studies suggested that domains 5 and 6 (D5 and D6) of group II introns act together in splicing and that the two helical structures probably do not interact by helix stacking. Here, we characterized the major Mg2+ ion- and salt-dependent, long-wave UV light-induced, intramolecular crosslinks formed in 4-thiouridine-containing D56 RNA from intron 5gamma (aI5gamma) of the COXI gene of yeast mtDNA. Four major crosslinks were mapped and found to result from covalent bonds between nucleotides separating D5 from D6 [called J(56)] and residues of D6 near and including the branch nucleotide. These findings are extended by results of similar experiments using 4-thioU containing D56 RNAs from a mutant allele of aI5gamma and from the group IIA intron, aI1. Trans-splicing experiments show that the crosslinked wild-type aI5gamma D56 RNAs are active for both splicing reactions, including some first-step branching. An RNA containing the 3-nt J(56) sequence and D6 of aI5gamma yields one main crosslink that is identical to the most minor of the crosslinks obtained with D56 RNA, but in this case in a cation-independent fashion. We conclude that the interaction between J(56) and D6 is influenced by charge repulsion between the D5 and D6 helix backbones and that high concentrations of cations allow the helices to approach closely under self-splicing conditions. The interaction between J(56) and D6 appears to be a significant factor establishing a side-by-side (i.e., not stacked) orientation of the helices of the two domains.