Calretinin and calbindin in the retina of the developing chick

Summary Calretinin and calbindin-D28k are two calcium-binding proteins that are present in largely different sets of nerve cells in the central nervous system. Their appearance during development of the chick retina was studied by immunohistochemistry and Western blots. The patterns are mature one day before hatching. Each cell type acquires its characteristic calcium-binding protein several days after its differentiation has started, but in most cases before morphological maturation is complete. There is also an early phase of calbindin immunoreactivity in many immature amacrine cells, and of calretinin immunoreactivity in the presumptive photoreceptor layer, suggesting that these proteins may have distinct functions in differentiating cells.Abbreviations CR+ Immunoreactive for calretinin only - CB+ immunoreactive for calbindin only - CR+CB+ immunoreactive for both antisera - IPL inner plexiform layer - OPL outer plexiform layer     

Structural analysis of the specific capsular polysaccharide of Streptococcus pneumoniae type 45 (American type 72)

The specific capsular polysaccharide of Streptococcus pneumoniae type 45 (American type 72) was found to be a high molecular weight polymer composed of D-galactose, 2-acetamido-2-deoxy-D-galactose, 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2-deoxy-L-fucose, L-rhamnose, glycerol, and phosphate (2:1:1:1:1:1:1). Partial hydrolysis, dephosphorylation, methylation analysis, periodate oxidation studies, and one- and two-dimensional 1H and 13C high-field nuclear magnetic resonance experiments showed the polysaccharide to be a branched polymer of a 1-phosphoglycerol-substituted hexasaccharide repeating unit having the structure: (formula; see text).     

Membrane proteins are critical targets in free radical mediated cytolysis

The hypothesis that proteins are critical targets in free radical mediated cytolysis was tested using U937 mononuclear phagocytes as targets and iron together with hydrogen peroxide to generate radicals. Those conditions which, after a lag of approx. 30 min, led to drastic lysis were also associated with very rapid membrane depolarisation. Conversely, when the early membrane depolarisation was prevented (by the addition of chelator and catalase), so was lysis. A similar correlation between early membrane depolarisation and subsequent lysis was also observed when the cells were exposed to a toxin from Actinobacillus actinomycetemcomitans. Those conditions of radical attack which led to lysis normally caused substantial lipid peroxidation. However, depolarisation and subsequent lysis were not prevented even when lipid peroxidation was completely suppressed by exogenous antioxidant. ATP levels were not grossly affected within the critical first 30 min period. These data exclude lipids and ATP as the target for lytic damage. We argue therefore that proteins are probably amongst the primary targets in cytolysis by radicals.     

Some aspects of the biology of the red bass, Lutjanus bohar (Forsskal), from the Tigak Islands, Papua New Guinea

The growth rate, food, reproduction and mortality rates of the red bass, Lutjanus bohar , are described for fish associated with coral reefs in the Tigak Islands of Papua New Guinea. Spawning intensity by month was investigated by examination of gonad maturation stages. The length–at–maturity of female L. bohar was estimated from the relationship between changes in gonad index with increasing size. Female fish matured at about 45–0 cm fork length. The Petersen method was used to obtain age–at–length data from length frequency data of L. bohar captured by hook–and–line fishing. The growth of L. bohar was described by a von Bertalanffy growth curve with the parameters AT=0.27 and L_∞ = 81.7 cm. There was a positive correlation between water depth (D) and the length of L. bohar (L) that was described by L = 318–8+ 3.050 D . The mortality rates ( Z ) of L. bohar from shallow (≤30m) and deep (≤80m) water were estimated from length converted catch curves and were 1–181 and 0–64 respectively. The high Z value for the shallow water population of L. bohar was ascribed to the migration of larger specimens into deeper water. The deep water population of L. bohar remains unfished and the estimate of Z was believed to be equivalent to the natural mortality rate ( M ). An empirical method of calculating M gave an estimate of 0.59. Lutjanus bohar is a carnivorous piscivore that feeds throughout the water column.     

Selective allele loss and interference between cauliflower mosaic virus DNAs

Summary Some cauliflower mosaic virus (CaMV) alleles are selectively lost during growth of the virus in mixedly infected turnip plants. Viral DNA from plants co-inoculated with DNA of the cabbage S isolate and infectious cabbage S DNA with an extra EcoRI restriciion site lacked the extra site. The EcoRI allele was also lost in most plants co-inoculated with a non-infectious mutant of cabbage S DNA while little selective allele loss was observed with two other non-infectious mutant DNAs. Plants co-inoculated with DNAs of closely-related isolates (CM4-184 and W) contained both parental viral DNAs and some DNAs with characteristics of both parents. Interference, scored as a reduced frequency of infection or a delay in symptom appearance relative to plants inoculated with wild-type DNA, occurred when plants were inoculated with wild-type and mutant DNAs covalently attached to one another in partial dimer plasmid DNAs. Similarities in the conditions leading to selective allele loss and those leading to interference suggest that both may have been due to active gene conversion between CaMV DNA molecules.     

1H-15N heteronuclear NMR studies of Escherichia coli thioredoxin in samples isotopically labeled by residue type

Ten samples of Escherichia coli thioredoxin were individually isotopically enriched by residue type via growth of an appropriate auxotrophic strain on media supplemented with one 2H, 15N-enriched amino acid. 1H observe-heteronuclear decoupling experiments were conducted on these samples making use of the 95-Hz 1H-15N amide J1 coupling. Subtraction of near-resonance from off-resonance 15N decoupled spectra generated difference patterns corresponding only to protons directly bonded to 15N nuclei. For the ten different enriched residue types observed to date, every labeled position (60) has been observed as a resolved resonance. The spectral dispersion in both the 1H and the 15N dimensions was roughly 1500 Hz (at 500-MHz field strength) with rather little apparent dependence on residue type. With the exception of the glycine-enriched sample, the range of the J1 coupling constants was not much greater than the precision of the measurements (1.5-2.0 Hz). However, for the glycine residues the J1 amide coupling values varied over a range of 10 Hz.     

Rapid determination of DNA synthesis in adherent cells grown in microtiter plates

A specific, rapid, and economical method for measuring the extent of DNA synthesis in adherent rat hepatoma H4-II-E cells grown in 96-well microtiter plates is described. The adherent cells were pulsed for 1 h with [methyl-3H]thymidine, released from the substratum by trypsinization, and collected on fiberglass filters with a MASH II cell harvester. The amount of radioactivity incorporated was directly proportional to the number of cells per well. Growth curves generated by measuring [methyl-3H]thymidine incorporation and counting the number of cells per well were identical. Experiments with inhibitors of DNA, protein, and RNA synthesis demonstrated that this method selectively measured DNA synthesis. In addition, [3H]thymidine uptake showed excellent correlation with autoradiographic assessment of DNA synthesis. This specific and sensitive method for determining DNA synthesis in microtiter cultures should facilitate studies of effects of various growth-controlling agents on epithelial, fibroblastic, and other cells which grow as adherent cells in culture.     

Chemically induced vasospasm: the effect of ischemia, vessel occlusion, and adrenergic blockade

Vasospasm in revascularized tissue can compromise tissue perfusion even though the microsurgical anastomoses remain patent. Circulating catecholamine stimulates peripheral vasoconstriction. Chemical vasospasm was induced by the intraarterial administration of norepinephrine to denervated rat hind limbs. Heel pad blood flow was assessed by laser-Doppler velocimetry. Mean blood flow was 463 +/- 106 in the denervated leg and 337 +/- 50 in the opposite (intact) leg (p less than 0.01). Flow in the denervated leg decreased 78 percent (463 +/- 106 to 100 +/- 23) within 5 minutes of norepinephrine administration and did not return to normal for 30 minutes. Norepinephrine administration in the presence of 1 and 3 hours of ischemia decreased flow at 5 minutes to 6.6 and 6.0 percent of normal, respectively (31 +/- 14, 28 +/- 14, control 100 +/- 23; p less than 0.001). Administration of intraarterial norepinephrine distal to a femoral artery occluded for 1 and 5 minutes reduced flow following clamp release to 11.2 and 7.1 percent of normal 5 minutes after clamp release (52 +/- 9, 33 +/- 7, control 100 +/- 23; p less than 0.001). Comparison of the 1-minute and 5-minute groups to each other showed a significant flow decrease in the 5-minute group (p less than 0.007). This indicates that the observed decrease in flow was related both to the presence of a vessel occlusion and to the length of the occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)