Secondary structure of mouse and rabbit α- and β-globin mRNAs: Differential accessibility of α and β initiator AUG codons towards nucleases

The nucleotide sequence from the 5′ terminus inward of one third of mouse α- and β^maj-globin messenger RNAs has been established. In addition, using 5′ ³²P end-labeled mRNAs as substrates and S1 and T1 nucleases as probes for single-stranded regions, the secondary structures of mouse and rabbit α- and β-globin mRNAs have been analyzed. Our results indicate that the AUG initiator codon in both mouse and rabbit β-globin mRNA is quite susceptible to cleavage with S1 and T1 nucleases, suggesting that it resides in a single-stranded exposed region. In contrast, the initiator AUG in the α-globin mRNA of both species is inaccessible to cleavage, indicating that it is either buried by tertiary structure or is base-paired. Since the rate of initiation of protein synthesis with β-globin mRNA in rabbit reticulocyte is 30–40% faster than for α-globin mRNA, these results imply a possible correlation between the differential rates of initiation with these two mRNAs and the accessibility of the respective AUG initiator codons.     

Uptake of the components of phenylalanylphenylalanine and maltose by intestinal epithelium

The observed rate of phenylalanine absorption into rat intestinal rings with 0.5 or 5.0 mM phenylalanine is greater than that for absorption of phenylalanine from 0.25 or 2.5 mM Phe-Phe, respectively. With the amino acid phenylalanine, V for absorption is the same whether Na⁺ is present (149 mM) or absent, but the concentration at which the half-maximal transport rate occurred (K_t) is greater in the absence of Na⁺. For Phe-Phe, the V decreases in the absence of Na⁺ whilst K_t is not influenced by the Na⁺ concentration. The different effect of Na⁺ on Phe and Phe-Phe transport indicates that the absorptive mechanism for Phe-Phe is different from that for phenylalanine. Absorption of a mixture of [U-¹⁴C]Phe-Phe and Phe-[G-³H]Phe showed identical rates of uptake of the carboxyl and amino terminal amino acids.Studies of transport of radioactive maltose showed that the rates of uptake of the reducing and non-reducing glucosyl moieties are identical. Radioactive maltose absorption is not inhibited by glucose oxidase.These results provide evidence that in intestinal epithelium, hydrolysis of Phe-Phe and maltose does not occur on the cell surface with release of the hydrolyzed products to the medium. Rather, hydrolysis and release of the reaction products occur at a point on the cytosol side of a diffusion barrier located in the brush border membrane.     

Inhibitors of superoxide dismutases: A cautionary tale

An extensive search resulted in the identification of pamoic acid as an inhibitor of superoxide dismutases. Pamoic acid appeared to rapidly and reversibly inhibit all types of superoxide dismutases and did so in both the cytochrome c reduction and in the dianisidine photooxidation assays, used to measure this activity. It could nevertheless be shown that pamoic acid did not at all inhibit superoxide dismutase but rather diminished the sensitivity of the assays. The mechanism proposed to account for this effect involved oxidation of pamoate, by O₂^?, to yield a pamoate radical which can then reduce cytochrome c or oxidize pyrogallol. Pamoate thus competes with superoxide dismutase for the available O₂^?, without affecting the observable effects of that O₂^? upon cytochrome c or upon pyrogallol. It consequently makes these assays less responsive to superoxide dismutase, while appearing to be without effect in the absence of superoxide dismutase. Several of the predicted consequences of this proposal were affirmed. Other workers, interested in finding inhibitors for superoxide dismutases, are hereby forwarned of this subtle snare.     

Effects of inorganic amendments (N,P and K) to soil on the rhizosphere microflora of antirrhinum plants infected withVerticillium dahliae Kleb

Summary A study of the inorganic amendments (N, P and K) to soil, and their effect on the rhizosphere microflora, as well as their relation to the control of wilt of antirrhinum plants caused byVerticillium dahliae Kleb. was done. Ammonium sulphate was the only chemical found to be significantly inhibitory toV. dahliae in vitro. Soil amendments (NPK) affected the rhizosphere microorganisms of the antirrhinum plants. Higher concentration of the chemicals were phytotoxic. It was further observed that ammonium sulphate, and the combined chemicals (NPK 25%) in soil delayed the senescence in healthy plants, suggests that chemical fertilisers affected the host plants directly. Addition of ammonium sulphate (0.25%), calcium nitrate (0.25%, 0.5%) combined NPK (0.25%) to soil caused considerable reduction in disease severity. It is assumed that this reduction may be caused by the (1) fungitoxic nature of the chemicali.e. ammonium sulphate, (2) antagonistic environment for the pathogen in the rhizosphere was boostedi.e. where calcium nitrate was added as soil amendments and (3) reduction in disease severity in soil-amended with combined NPK, may be due to the fact that antagonistic actinomycete population was boosted in the rhizosphere.     

Effects of N-serve (2-chloro-6-(trichloromethyl)pyridine) formulations on nitrification and on loss of nitrate in sand culture experiments

Summary N-serve (2-chloro-6-(trichloromethyl)pyridine) was tested as an inhibitor of nitrification of ammonium or urea in sand cultures. Nitrification was reduced but not prevented by N-Serve present at between 5 and 20 ppm in solution or by weight of sand. In the presence of root debris and acetone, used in some experiments at 2–4 ml/l of nutrient to convey N-Serve, denitrification was stimulated under the same conditions and resulted in loss of a large proportion of nitrate, probably mainly as gaseous products and some nitrite. These losses were greater when N-serve was also present. There was also conversion of nitrate to an insoluble form in the sand. A smaller proportional loss of nitrate occurred in other treatments in the presence of root debris when N-Serve was added without acetone, either as the commercial formulation 24E or as a solid. Thus, using N-Serve to inhibit nitrification may encourage denitrifying organisms especially in the presence of carbon sources including root debris or acetone. Large decreases of nitrate reductase activity in plants produced by using N-Serve in the presence of ammonium or urea were caused as much by losses of nitrate in the presence of acetone as by prevention of nitrate formation. Other N-Serve treatments (solid or 24E) decreased enzyme induction by between 50 and 90 per cent as a result mainly of reduced nitrification.     

Effect of neutralization and addition of urea,sucrose, and various glycols on phosphorus absorption and leaf damage from foliar-applied phosphate

Summary Inclusion of sucrose in the solution applied to soybean (Glycine max L. merr.) leaves much reduced the severity of the damage to the leaves from application of urea and, to a lesser extent, from application of phosphorus (P) as orthophosphoric acid. Sucrose had no evident effect on P absorption. Damage to the leaves from joint application of orthophosphoric acid and urea exceeded the sum of the damage caused by the substances individually. Urea did not seem to influence P absorption, but the effect, if any, was not readily determined because nearly all values for P absorption exceeded 90%.Neutralization of orthophosphoric acid with nitrogen-containing organic bases, including choline, guanidine, and guanyl urea, did not prove useful as a technique for increasing the quantity of orthophosphate that could be applied without damage to the leaves.Absorption and translocation of orthophosphate by corn (Zea mays L.) and soybean leaves were not influenced by the pH of the solution within the range from 2 to 10. Absorption of tripolyphosphate by corn leaves decreased with an increase in pH of the solution applied, but translocation of the absorbed P was not influenced by pH. With soybeans, absorption of tripolyphosphate decreased with an increase in pH of the solution. Translocation of P applied to soybean leaves as tripolyphosphate was less than 5% of the amount absorbed within the first 24 hr and decreased with an increase in pH after 10 days.     

Stereospecificity of peptide transport by germinating barley embryos

The stereospecific requirements for peptide transport in the scutellum of germinating barley (Hordeum vulgare) embryos are described. Replacement of an L-amino acid residue in a peptide by its D-stereoisomer decreases the affinity of the peptide for the transport site, leading to a reduction in transport. Substitution of a second D-residue reduces affinity still further. The extent to which transport is inhibited depends upon the position of the D-residue in the primary sequence, with D-residues at the C-terminus of the peptide having the greatest effect. Competition between D- and L-peptides indicates that they both enter via the same transport system. Although D-amino acids can be accumulated when presented as a peptide, these same D-residues are not transported when supplied as the free amino acids. L-Leu-D-leu is accumulated intact against a concentration gradient, indicating the operation of an active transport mechanism that can function without the involvement of peptidase activity.     

Effector cells which mediate antibody-dependent cell-mediated cytotoxicity. II. Ontogeny of effector activity in murine spleen.

The specificity of T cells for syngeneic target cells is directed to both antigens and products of the major histocompatibility complex (MHC) on the target cell surface. This dual requirement is best accounted for by the altered-self hypothesis, which implies that the MHC products on a cell's surface are able to form complexes with many other proteins on the surface of the same cell. To account for the ability of MHC products to bind so many different cell surface antigens we propose that interactions in general among macromolecules on the surface of a membrane may be dramatically enhanced by a purely physical effect. This effect derives from the confinement of membrane macromolecules to an effective volume which is the product of membrane surface area times d, the distance over which the center of mass of the molecules can move in a vertical direction (perpendicular to the membrane surface). Because d is very small the effective concentrations of surface molecules are extremely high and their interactions are correspondingly enhanced.     

The heterocytotoxicity of human serum. II. Role of natural antibody and of the classical and alternative complement pathways.

Mouse lymphoid cells and tumor cell lines were employed as target cells for the investigation of the mechanism of heterocytotoxicity in human serum samples. It was shown that the heterocytotoxic effects were due to two differing mechanisms. Cytotoxicity was mediated in part, by activation of the alternative complement pathway on target cell membrane, a process which was antibody-independent. A second mechanism of cytotoxicity was induced by natural antibodies to a target cell, which probably mediated activation of the classical complement pathway. These data may shed light on the frequently observed cytotoxicity in mammalian sera for various target cells.     

Effect of 2450 MHz microwave exposure on behavioural thermoregulation in mice

1. 1.|The purpose of this study was to determine the threshold specific absorption rate (SAR) during exposure to 2450 MHz continuous wave (CW) microwaves that affected thermoregulatory behaviour in mice.

2. 2.|A Plexiglas shuttle box was placed inside a waveguide imposed with a temperature gradient. The temperature gradient allowed the mice to select a particular section of the shuttle box which was, presumably, related to their state of thermal comfort. Exposing the mice to 2450 MHz inside the waveguide at SARs of 0–5.3 W kg⁻¹ for 1 h caused no significant change in their preferred ambient temperature.

3. 3.|Increasing SAR from 5.3 to 18.1 W kg⁻¹ caused the animals to shift their position to the cooler end of the shuttle box.

4. 4.|Following termination of microwave exposure animals that had selected a cool ambient temperature returned to the warm side of the shuttle box.

5. 5.|It is concluded that for mice exposed to radiation at 2450 MHz the thermoregulatory behaviour is significantly affected at SARs of 5.3 to 9.9 W kg⁻¹.