Control of locomotion in vitro: I. Deafferentation

We previously described the ability to induce adult-like, coordinated airstepping following electrical stimulation of the brainstem in the hindlimb-attached, in vitro brainstem-spinal cord preparation. These findings suggest the presence at birth of supraspinal systems capable of activating and modulating spinal locomotor mechanisms, which presumably also are present at birth. The current study employed the hindlimb-attached in vitro brainstem-spinal cord preparation from 0- to 4-day-old rats maintained in oxygenated artificial cerebrospinal fluid. After the control threshold-frequency relationship for eliciting airstepping was established, the dorsal roots to the attached limbs were severed and the procedure was repeated. No changes in electrical threshold or major differences in the elicited locomotor pattern were observed after deafferentation, although the amplitude of the electromyograms decreased. The mean frequency of alternation at threshold before deafferentation was similar to that after deafferentation. However, the maximum mean frequency induced by suprathreshold stimulation was significantly higher after deafferentation than that before deafferentation. These results suggest that (1) the supraspinal modulation of spinal locomotor mechanisms is not entirely dependent on afferent input; (2) intrinsic spinal locomotor mechanisms are present in the spinal cord at birth; and (3) afferent input may limit the maximum frequency of alternation of the limbs early in development.     

Structures of paralytic acylpolyamines from the spider Agelenopsis aperta

The structures are given for five paralytic acylpolyamines from the venom of the funnel web spider, Agelenopsis aperta. The acyl moieties are derived from (3-indolyl)acetic acid, (4-hydroxy-3-indolyl)acetic acid, and 4-hydroxybenzoic acid. The polyamine portions of the toxins are novel. Three toxins (AG489, AG505, and AG452) contain 1, 5, 9, 13, 18, 22-hexaazadocosane which is unique as a natural polyamine because of its length and hydroxylation at the 5-aza position. The polyamine portions of two other alpha-agatoxins (AG488 and AG504) are unusual also, containing guanidinooxy moieties.     

Studies on the Cambodian I strain of Plasmodium falciparum in Aotus monkeys

The Cambodian I strain of Plasmodium falciparum, originally from Kampuchea was adapted for development in three different types of Aotus monkeys. High-level parasitemias were readily produced in splenectomized Colombian A. trivirgatus griseimembra monkeys. Initially, only minimal parasitemias developed in A. t. trivirgatus monkeys from Colombia. However, in one animal, adaptation occurred and high-level parasitemias were obtained during the second recrudescence of the infection. Passage to other A. t. trivirgatus monkeys indicated that the parasite was well adapted for development in splenectomized animals; low to moderate parasitemias were still produced in intact animals. This line of the parasite produced high level parasitemias when inoculated into splenectomized Aotus monkeys from Peru. Infections in Anopheles freeborni mosquitoes were obtained as late as the 7th passage in A. t. griseimembra monkeys and as late as the 7th recrudescence of the infection in an individual monkey (348 days after inoculation). The sporogonic cycle was completed in An. freeborni mosquitoes, and one transmission to an A. t. griseimembra monkey via the bites of infected mosquitoes was obtained.     

The productivity of pine plantations in relation to previous land use

Summary Differences in the productivity of establishedP. radiata plantations on pasture and forest soils were found to be reproducible withP. radiata seedlings in a glasshouse environment. The growth of seedlings on pasture soil exceeded that of seedlings on native forest soil (the ‘primary pasture effect’). After a history of pine on both native and pasture soils a residual effect of pasture on seedling growth was evident (the ‘secondary pasture effect’). However, the effect of a history of pine plantation (the ‘pine effect’) was to decrease the productivity of both native and pasture soil as assessed by seedling growth. These effects were not related to changes due to land management in mycorrhizal infection or in soil structure. The analysis of seedling growth leads to the conclusion that soil fertility, particularly the availability of nitrogen and phosphorus, has changed.     

Histopathogenesis of the Galls Induced by Nothanguina phyllobia in Solanum elaeagnifolium

The histopathogenesis of the foliar galls induced by Nothanguina phyllobia Thorne in Solanum elaeagnilolium Cav. was examined via serial sections prepared from plant shoots at 11 time intervals (0.5-30 days) following inoculation. Nematodes infected the blades and petioles of young leaves surrounding the shoot apex. Hypertrophy and hyperplasia of the palisade, pith, cortical, and vascular parenchyma resulted in the formation of confluent leaf, petiole, and stem galls up to 25 cm³ in volume. Externally, leaf galls were irregular, light-green, convoluted spheroid bulges distending the abaxial surface. Mature galls contained a cavity lined with parenchymogenous nutritive tissue comprising intercellular spaces and actively dividing hypertrophied cells. These cells contained granular cytoplasm, hypertrophied nuclei, and brightly stained large nucleoli. Vascular tissues were not discernibly affected during the early stages of gall development. As gall development progressed, however, vascular elements were often displaced and disoriented. The histopathology of this nematode indicates that N. phyllobia is a highly specialized parasite and, for that reason, is suitable as a biological control agent.     

Production rates of prostaglandin F, 6-keto-PGF1α and thromboxane B2 by perifused human endometrium

Human endometrium obtained from fresh hysterectomy specimens was perifused for 7 hr in 95% O₂/5% CO₂ at 37°C. The phase of the menstrual cycle was determined by histological examination. The concentrations of PGF, 6-keto-PGF_1α and TxB₂ in 20 min fractions of the perifusion medium were measured by radioimmunoassay and production rates were calculated in terms of dry weight of tissue. Biphasic patterns of production were observed; high initial values fell to about 20% at 2 hr and then increased to relatively stable values at about 4 hr which were maintained for the next 2 hr. During this latter period, production rates in endometria taken at different phases of the cycle differed markedly from each other; the production rates of PGF in secretory and early proliferative endometria were low (15.8 ± 2.6, mean ± SEM and 67.2 ± 8.3 ng/min/g respectively) whereas they were high in late proliferative and premenstrual endometria (188.0 ± 16.7 and 196.4 ± 16.9 ng/min/g respectively). The patterns of production of 6-keto-PGF_1α and TxB₂ were similar to those of PGF but the absolute values were much lower (<10%). We conclude that the observed rates of production of prostaglandins by perifused human endometrium are consistent with synthesis being stimulated either by estrogen or withdrawal of hormonal support and being inhibited by progesterone.     

The North American «Hipparion Datum and implications for the Neogene of the Old World

A large sample of skulls and associated dentitions of Cormohipparion sphenodus (Cope, 1889). new combination, is described from Miocene deposits of Nebraska, Colorado, New Mexico, and California. This species is morphologically more advanced than the most primitive species of the genus, C. goorisiMacFadden & Skinner, 1981, and more primitive than a third and more advanced species of this genus. C. occidentale (Leidy, 1856). In several characters such as configuration of the preobrital fossa, size, hypsodonty, shape of protocone, and complexity of enamel plication, C. sphenodus is morphologically similar, and potentially related to, some Old World Vallesian «Hipparion. In addition, based on radiometric calibration of stratigraphic sequences in Nebraska and California, C. sphenodus spans an interval of time from about 13.6 to 11 Ma. Inasmuch as the Old World «Hipparion Datum is calibrated at about 12 Ma, we conclude that C. sphenodus exhibits the morphological and chronological prerequisites to have been directly ancestral to some of these Old World forms.The hipparion horses apparently were a polyphyletic assemblage of several genera, and their biogeographic and biochronologic history is more complex than classically thought. Several dispersal events involving these horses may have occurred between the New and Old Worlds during the late Miocene.     

Catabolism of 3- and 4-hydroxyphenylacetate by the 3,4-dihydroxyphenylacetate pathway in Escherichia coli.

Various strains of Escherichia coli (but not strain K-12) were found to grow on 3-hydroxyphenylacetate and 4-hydroxyphenylacetate. Both compounds were catabolized by the same pathway, with 3,4-dihydroxyphenylacetate as a substrate for fission of the benzene nucleus, and with pyruvate and succinate as products. All the necessay enzymes were demonstrated in cell extracts prepared from induced cells but were essentially absent from uninduced cells. Mutants unable to grow on 3- and 4-hydroxyphenylactetate were defective in particular enzymes of the pathway. The characteristics of certain mutants indicated that either uptake or hydroxylation of 3- and 4-hydroxyphenylacetate may involve a common protein component. E. coli also grew on 3,4-hydroxyphenylacetate, with induction of the enzyme necessary for its degradation but not those for the uptake-hydroxylation of 3- and 4-hydroxyphenylacetate.     

Purification, properties and kinetics of sheep and human renin substrates.

Sheep plasma renin substrate was purified 1,200-fold by using nephrectomised sheep plasma, followed by DEAE-Sephadex chromatography and gel filtration. The purified substrate contained 8 mu-g angiotensin II/mg protein and had an estimated molecular weight of 52,000. The kinetic characteristics of the purified substrate were identical both to those of unpurified nephrectomised sheep plasma and to normal sheep plasma substrates. At pH 7.5, K-m of the human renin-sheep substrate reaction was 0.29 mu-M and for sheep renin-sheep substrate, 2.0 mu-M. Sheep substrate was susceptible to peptic digestion with generation of pepsitensin. Human renin substrate was less readily purified. DEAE-Sephadex chromatography of plasma from pregnant women at 36-40 weeks' gestation produced a 70-fold increase in purity (0.9 mu-g angiotensin II/mg protein). No further increase was achieved with gel filtration. Human renin substrate behaved as a larger (mol. wt. 82,000) more anionic protein than sheep substrate and was resistant to the proteolytic actions of both pepsin and sheep renin. K-m for the human renin-human substrate reaction was high and could not be accurately determined (range 3-8 mu-M, mean 5.7 mu-M). The presence of human substrate in a human renin-sheep substrate system did not alter the measured initial velocity. In both sheep and man, the normal concentration of renin substrate is considerably less than K-m and must therefore be considered a determinant of angiotensin production rate in vivo.     

A 25,000 molecular weight protein constituent of human amyloid fibrils related to amyloid protein AA

Amyloid fibrils from a patient with diffuse amyloid disease are dissociated in 6 m guanidine hydrochloride and fractionated by gel chromatography. Two major components are separated on Sepharose 6B. Both proteins are characterized by chromatography, immunodiffusion, discontinuous gel electrophoresis, amino acid tryptic peptide mapping and amino acid sequence analysis. The smaller of the two components is typical of the known protein AA by size (8400 daltons), amino acid composition and a 30-residue N-terminal sequence. The larger of the components (25,000 daltons) undergoes electrophoresis as a single band and appears unaffected by thiol reduction. It differs from protein AA in amino acid content and by its tryptic peptide map, although it contains an N-terminal amino acid sequence identical to protein AA when carried to 20 residues. Treatment of this larger component by mild acid hydrolysis results in the release of the 8400-dalton protein AA. Fractionation after guanidine hydrochloride treatment of this particular amyloid fibril preparation is compared to the fractionation of a typical secondary amyloid preparation that contains only protein AA as the major component. The origin and relationship of the 8,400- and 25,000-dalton protein components is discussed.