Comparison of glucokinase in C3H/He and C58 mice that differ in their hepatic activity

Partially purified preparations of the hepatic glucokinase from C3H/He and C58 inbred mice have been used to explore the molecular basis for the observed twofold difference in activity between the strains. The single codominant gene that appears to regulate activity, the alleles of which are designated Gk^a and Gk^b, respectively, for the two strains, could represent a structural gene change. This now seems unlikely because the mouse enzyme, although showing small differences from rat glucokinase, appeared to be identical in the two strains with respect to thermal stability, electrophoretic mobility in agarose gels, and kinetic properties such as the apparent K _m values for MgATP^2– and glucose and the unique cooperative interaction with the latter substrate. The enzymes also reacted identically in a range of immunological tests (double-diffusion, immunoelectrophoresis, immune precipitation and immune inhibition assays) and ELISA immune inhibition assays indicated that the twofold difference in activity was due to a similar difference in antigenically active enzyme. Genetic control over the physiologically significant regulation of enzyme amount is therefore probable.This work has been supported in part by a grant from the British Diabetic Association and a Training Studentship to PAJ from the Medical Research Council (U.K.).     

Adaptation to High-Intensity, Low-Wavelength Light among Surface Blooms of the Cyanobacterium Microcystis aeruginosa

Natural populations of the nuisance bloom cyanobacterium Microcystis aeruginosa obtained from the eutrophic Neuse River, N.C., revealed optimal chlorophyll a-normalized photosynthetic rates and resistance to photoinhibition at surface photosynthetically active radiation (PAR) intensities. At saturating PAR levels these populations exhibited higher photosynthetic rates in quartz than in Pyrex vessels. Eucaryotic algal populations obtained from the same river failed to counteract photoinhibition. At saturating PAR levels, such populations generally yielded lower photosynthetic rates in quartz containers than they did in Pyrex containers. Cultivation of natural Microcystis populations under laboratory conditions led to physiologically distinct populations which had photoinhibitory characteristics similar to those of other cultured cyanobacterial and eucaryotic algae. Our findings indicate that (i) photosynthetic production among natural surface populations is best characterized and quantified in quartz rather than Pyrex incubation vessels; (ii) extrapolation of natural photoinhibitory trends from laboratory populations is highly subjective to culture and PAR histories and may yield contradictory results; and (iii) buoyant surface-dwelling populations, rather than exhibiting senescence, are poised at optimizing PAR utilization, thereby maintaining numerical dominance in eutrophic waters when physico-chemical conditions favor bloom formation.     

Preliminary crystallographic data for cross-linked (lysine7-lysine41)-ribonuclease A

A cross-linked derivative of ribonuclease A, N^ε,N^ε′-(2,4-dinitrophenylene-1,5)-(lysine⁷-lysine⁴¹)-RNase A, has been crystallized by dialysis against 30% ($\text{v}\text{v}$) ethanol/water mixtures buffered at high pH. Single crystals belong to the orthorhombic space group P2₁2₁2₁, $\text{a = 37.2}\text{A}\text{?}\text{, b = 41.2}\text{A}\text{?}\text{, b = 41.2}\text{A}\text{?}$, with one molecule in the Crystallographic asymmetric unit.     

Potato Lectin: A Cell-Wall Glycoprotein

The activity and the amount of potato lectin were measured inpotato tuber slices (Solanum tuberosum cv. Huinkul) aeratedfor 48 h. Lectin was found in a soluble form, liberated to themedium and associated with insoluble structures. Polyacrylamidegel electrophoresis in denaturating conditions and immunologicaltechniques indicated that the lectins associated to cell wall,soluble or liberated to the medium, were identical. The cell-wallfraction was found to contain 65% of total lectin in the tuber.The possible role of potato lectin in tubers was discussed. (Received June 5, 1985; Accepted September 3, 1985)     

The cytoskeletal architecture of interdigitated microvilli in the photoreceptors of some nocturnal spiders

Summary The photoreceptor microvilli of some nocturnal spiders (Isopeda andOlios in theSparassidae, andClubiona in theClubionidae) are wide (80–140 nm), and microvilli from adjacent receptors are interdigitated. Because microvillar diameters are relatively large in relation to the thicknesses of thin sections, it is possible to examine cytoskeletal structures closely associated with the microvillar plasmalemmae directly.Retinae were treated with a specific inhibitor of cysteine proteases before primary fixation for electron microscopy in a Ca²⁺-chelating medium. Cytoskeletal components were stabilized with tannic acid. A variety of microvillar profiles was obtained, consistent with an assumption that they represent imperfect preservation of anin vivo plasmalemmal undercoat, inferred to consist of longitudinally-disposed microfilaments, presumptively bonded to the microvillar plasmalemma. The microvillar lumen is inferred to be empty of cytoskeletal components in life.This model is discussed in terms of 1. the cytoskeletal organisation of microvilli of the primitive photoreceptors of a leech (Blest et al. 1983), where the arrangement of microfilaments resembles that in the vertebrate intestinal brush-border; 2. the large complement of membrane-associated oligomeric actin in rhabdoms of crayfish, where identifiable microfilaments cannot be resolved within microvilli by transmission electron microscopy (de Couet et al. 1984), and a single visualizable axial filament of uncertain composition is linked to the plasmalemma by side-arms.     

Heterologous hybridization of Azospirillum DNA to Rhizobium nod and fix genes

Abstract Probes containing the nod and hsn regions of Rhizobium meliloti and the fixABC genes of Rhizobium japonicum were used to perform hybridization experiments with endonuclease-restricted DNA from Azospirillum brasilense strains and 2 Azospirillum lipoferum strains. Homology to nod, hsn and fixA was found in the 4 Azospirillum strains.     

Reduction in headache patients' medical expenses associated with biofeedback and relaxation treatments

Comparisons are made of self-reported medical costs from a sample of headache patients who underwent various combinations of relaxation training and biofeedback training. The average costs for the 2 years prior to self-regulatory treatment were $955±480 (3 SEM) for 45 patients; for the 2 years after completing treatment the average costs were $52±28 (3 SEM) for patients. Within the limitations of the study, medical costs do seem to have been markedly reduced.This research was supported by a grant from NINCDS, NS-15235.     

Stoichiometric binding of diacylglycerol to the phorbol ester receptor

The major phorbol ester receptor is the Ca++-activated, phospholipid-dependent protein kinase C. Diacylglycerol stimulates protein kinase C in a fashion similar to the phorbol esters. Likewise, it inhibits phorbol ester binding competitively. Both results suggest that diacylglycerol is the/an endogenous phorbol ester analogue. Alternatively, the diacylglycerol might simply be acting to modify the phospholipid environment of the protein. If diacylglycerol were indeed functioning as an analogue, it should interact with the receptor stoichiometrically. This interaction can be quantitated by measuring the perturbation in apparent diacylglycerol binding affinity as a function of the ratio of diacylglycerol to receptor. We report here that 1,2-dioleoylglycerol interacts with the receptor with the predicted stoichiometry.     

Sperm aging in the male after sexual rest: Contribution to chromosome anomalies

The chromosome complement was studied in first-cleavage metaphases of mouse zygotes resulting from sperm aged in the male physiologically, after sexual rest. Females were inseminated by control males mating at 3-day intervals while experimentals mated to males that had had a sexual rest of 14 or more days. A total of 1954 eggs were collected 33–35 h post-HCG from 101 superovulated females mated to 42 controls and 43 experimental males. The fertilization rate was similar in both groups, being 84% and 85%, respectively. G-banded or Q-banded chromosomes were analyzed in 301 (68.3%) controls and 392 (49%) experimental first-cleavage metaphases. The overall rate of chromosome anomalies in controls was 4.45% as compared to 10.94% in experimentals, a highly significant difference. In the experimental group compared to controls, the frequency of trisomy, triploidy, structural rearrangements, and tetraploidy increased from 3.9% to 6.9%, 0% to 1.6%, 0.8% to 2.8%, and 0% to 1.3%, respectively. The genomic source of origin of the abnormalities was determined on the basis of differential condensation of the genomes. In the experimentals, grossly unbalanced sperm (diploids, disomics, double disomics, and those with large fragments) fertilized significantly more oocytes compared to controls. Our results implicate an advantage either in numbers or fertilizing capability for chromosomally abnormal sperm in a physiologically aged population.