Role of DNA in the activation of the Cry1A insecticidal crystal protein from Bacillus thuringiensis

The Cry1A insecticidal crystal protein (protoxin) from six subspecies of Bacillus thuringiensis as well as the Cry1Aa, Cry1Ab, and Cry1Ac proteins cloned in Escherichia coli was found to contain 20-kilobase pair DNA. Only the N-terminal toxic moiety of the protoxin was found to interact with the DNA. Analysis of the crystal gave approximately 3 base pairs of DNA per molecule of protoxin, indicating that only a small region of the N-terminal toxic moiety interacts with the DNA. It is proposed that the DNA-protoxin complex is virus-like in structure with a central DNA core surrounded by protein interacting with the DNA with the peripheral ends of the C-terminal region extending outward. It is shown that this structure accounts for the unusual proteolysis observed in the generation of toxin in which it appears that peptides are removed by obligatory sequential cleavages starting from the C terminus of the protoxin. Activation of the protoxin by spruce budworm (Choristoneura fumiferana) gut juice is shown to proceed through intermediates consisting of protein-DNA complexes. Larval trypsin initially converts the 20-kilobase pair DNA-protoxin complex to a 20-kilobase pair DNA-toxin complex, which is subsequently converted to a 100-base pair DNA-toxin complex by a gut nuclease and ultimately to the DNA-free toxin.     

Recombinant human acid alpha-glucosidase: high level production in mouse milk, biochemical characteristics, correction of enzyme deficiency in GSDII KO mice

Glycogen storage disease type II (GSDII) is caused by lysosomal acid alpha-glucosidase deficiency. Patients have a rapidly fatal or slowly progressive impairment of muscle function. Enzyme replacement therapy is under investigation. For large-scale, cost-effective production of recombinant human acid alpha-glucosidase in the milk of transgenic animals, we have fused the human acid alpha-glucosidase gene to 6.3 kb of the bovine alphaS1-casein gene promoter and have tested the performance of this transgene in mice. The highest production level reached was 2 mg/ml. The major fraction of the purified recombinant enzyme has a molecular mass of 110 kDa and resembles the natural acid alpha-glucosidase precursor from human urine and the recombinant precursor secreted by CHO cells, with respect to pH optimum, Km, Vmax, N-terminal amino acid sequence and glycosylation pattern. The therapeutic potential of the recombinant enzyme produced in milk is demonstrated in vitro and in vivo. The precursor is taken up in a mannose 6-phosphate receptor-dependent manner by cultured fibroblasts, is converted to mature enzyme of 76 kDa and depletes the glycogen deposit in fibroblasts of patients. When injected intravenously, the milk enzyme corrects the acid alpha-glucosidase deficiency in heart and skeletal muscle of GSDII knockout mice.     

Manganese reduces myocardial reperfusion injury on isolated rat heart

It has been shown that reactive oxygen species produced during the early phase of myocardial post-ischemic reperfusion are one of the main causes of reperfusion injury. This observation has led to various antioxidant strategies using many reactive oxygen species scavengers, including manganese complexes. The aim of the present work was to provide a reference study of the effects of manganese itself (MnCl2) on isolated rat hearts submitted to global total normothermic ischemia (30 min) and reperfusion (60 min). McCl2 was administered either during the first 10 min reperfusion (10(-5)M and 10(-4)M) or throughout reperfusion (10(-4)M). After 10 min reperfusion, no functional difference was evidenced between control and manganese-treated groups, whereas high energy phosphate contents were significantly higher in treated groups. MnCl2 10(-4)M enhanced the recovery of developed pressure between 40 and 55 min reperfusion. At the end of reperfusion, hearts treated during the first 10 min reperfusion showed a better metabolic recovery. The group treated throughout reperfusion showed a better metabolic recovery, but a reduced coronary flow and a weak recovery of developed pressure. These results suggest that MnCl2, administered during the early phase of reperfusion, protects against myocardial reperfusion injury. This effect might be mediated by manganese antioxidant properties.     

Two roads to coma: the Scottish hypothesis

Clinical experience seems to indicate two separate types of entry to coma. Some patients follow a pathway characterized by confusion, hallucinations, mumbling delirium, myoclonic jerks, and seizures. The author has called this sequence of symptoms the high road to coma and hypothesized that its basic underlying pathophysiology involves increased neuronal firing rates. Other patients develop somnolence, lethargy, obtundation, and unresponsiveness without seizures or muscle twitches. This low road to coma involves either anatomical compression of the midbrain reticular formation or a metabolic or toxic disorder characterized by membrane stabilization and decreased neuronal excitability.     

Multiple infection with Toxocara canis. Influence of antihistamines and corticosteroids on the histopathological response

The histopathology of the tissues of mice infected with ten doses of 100 embryonated eggs of Toxocara canis and the influence of antihistamines and corticosteroids on these findings is described. The main lesions were confined to the liver and lungs. In infected but untreated control mice hepatic lesions consisted of periportal infiltrates, widespread foci of liver cell necrosis and polymorphonuclear and mononuclear accumulation and a few fibrotic granulomata. Treatment with antihistamines did not change these findings significantly, but corticosteroids decreased the amount of cellular infiltration and the size of the lesions. Pulmonary lesions consisted mostly of confluent infiltrates of polymorphonuclears, eosinophils and mononuclear cells with some macrophage proliferation. Neither drug seemed to affect the lung involvement.     

The effects of self-induced mood states on behavior and physiological arousal

The effects of reading emotionally loaded statements on behavioral tasks and physiological measures were investigated. Statements were constructed to arouse elation, depression, or neutrality. Ss were both pre- and posttested on Writing Speed, Reaction Time, Decision Time, and Spontaneous Verbalizations. Base rates were obtained for heart rate and galvanic skin response. Elation Ss significantly outperformed both Neutral and Depression Ss on the Reaction Time task. Scores for Neutral Ss fell between those of Elation and Depression Ss on three of the four behavioral measures. No significant differneces were found on the physiological measures.     

Cardiovascular and neuromuscular effects of dimethyl sulfoxide in anesthetized rabbits

In rabbits anesthetized with pentobarbital, the carotid arterial blood pressure and bilateral contractions of the gastrocnemius muscles due to electrical stimulation of the sciatic nerves were recorded. Intravenous administration of up to 1 ml of dimethyl sulfoxide/kg caused profound hypotension and eventually failure of neuromuscular transmission. Caution must be used in considering dimethyl sulfoxide as a solvent for drug administration.     

Influence of environment on passive immunity in calves

Passive immunity in neonatal calves is influenced by environment. Placing newly born Holstein calves (108 head) in three different housing environments (shade, cooled shade, hutch) during hot weather produced differences in body temperature, serum cortocosteroids, immunoglobulin IgG1 concentrations, and mortality. Experimental design permitted examination of effects due to treatments, time, differences in colostrum, and climatic environment in an analysis of variance. Calves exposed to the hotter, less desirable environment responded by having a higher mortality, higher serum corticosteroid concentration, and lower serum immunoglobulin IgG1 at 2 and 10 days after birth. All of these were correlated. Calves that died had serum immunoglobulin IgG1 falling below the mean for all experimental calves.