Ganglioside changes associated with temporal lobe epilepsy in the human hippocampus

To understand better the molecular and cellular events associated with status epilepticus, a multifaceted analysis has begun on hippocampal tissues therapeutically removed from patients with temporal lobe epilepsy. In this first study, quantitative changes in major ganglioside species are reported, as well as the immunocytochemical localization on the ganglioside GD3 in epileptic human hippocampus. Although significant variations were found between patients, the pattern of change was consistent when compared to normal values obtained from an autopsied specimen and the literature. Total ganglioside content was reduced in epileptic hippocampi, which was attributable, in part, to pyramidal cell loss found in CA1 and CA3. In each case, the percentage of ganglioside GD3 was increased significantly, while ganglioside GD1a decreased. The former change is probably associated with reactive astrocytosis and the latter with loss of neuronal dendrites. Immunocytochemical localization revealed GD3 in the stratum radiatum and the subgranular layer of the dentate gyrus. In these areas, GD3 was present in punctate structures and astrocytes. These findings indicate that GD3 increases in selected areas of the sclerotic hippocampus and is presumably related to localized accumulation of reactive glial cells. Since gangliosides have a high affinity for calcium and localized increase in extracellular calcium could disrupt normal neuronal function, the localized increase in GD3 may not only denote reactive glial cells but may contribute directly to the altered, hyperexcitable condition of epilepsy.     

Differential adhesivity of neuroepithelial cells and pioneering circumferential axons

To determine the initial growth pattern of pioneering axons and investigate the factors that may influence their guidance, the lateral margin of a stage 16+ chick brachial spinal cord was examined in serial thin sections. The specimen was prepared with hypertonic fixative which partially shrank the tissue and increased extracellular space. The retention of surface contact after shrinkage was used as an index of the relative adhesiveness between cells in situ. Six axons and growth cones were found within the reconstructed tissue; five were oriented dorsoventrally and one apparent motor neuron growth cone was oriented radially. The five circumferential axons originated from presumptive interneurons distributed in a dispersed pattern along the neural tube lateral wall. Four terminated with growth cones, and each extended a short distance (less than 30 microns) ventrally along the outer margin. No contact was found between these nonfasciculating axons or growth cones. Thus, the earliest intracentral axons appear to grow dorsoventrally from the outset with no appreciable wandering. Morphological features that may indicate their mechanism of guidance, including preformed cellular guides, extracellular channels, contact with basal lamina, and intercellular junctions were not found. The preferential retention of surface contact between adjacent endfeet, as well as between pioneering circumferential axons and neuroepithelial cells, suggests that these particular surfaces are mutually adherent. These findings are consistent with a proposed dorsal-to-ventral adhesive gradient mechanism of circumferential axonal guidance.     

Survival and transport of bacteria in egg washwater.

An evaluation of methods for monitoring the quality of water used to wash eggs at grading stations was undertaken to improve maintenance of bacterial viability during overnight sample transport. Bacterial content of samples at analysis would then better reflect conditions at the time eggs were washed. The interactive effects of temperature and the highly alkaline water conditions upon viability were the subjects of this study. Nine transport methods were examined for their efficacy in recovering total and coliform bacteria from recycled water used to wash eggs, and these were compared with samples analyzed at two commercial egg grading stations. Samples were shipped under test to the laboratory for analysis the following day. The survival of Staphylococcus aureus and Escherichia coli was also examined, but in a synthetic washwater matrix under various combinations of temperature (6 to 32 degrees C) and pH (9.5 to 10.5) to determine whether there was likely to be a different response to variations in transport treatment among gram-positive and -negative bacteria. S. aureus was much more resistant to the lethal effects of high pH and moderate temperature than E. coli. These results indicated that samples of high pH should be held (transported) at less than or equal to 13 degrees C to optimize bacterial survival. Considering cost, ease of manipulation, and the ability to protect both coliforms and the bacterial population as a whole, the method of choice for transport of industrial samples was the direct addition of washwater to containers in which powdered KH2PO4 and Na2S2O3 had been placed to yield final concentrations, when dissolved, of 0.2 and 0.05% (wt/vol), respectively.     

Strain and sex differences in the response of mice to drugs that induce protoporphyria: role of porphyrin biosynthesis and removal

A hepatic green pigment, inhibitory toward ferrochelatase, has been isolated from the liver of mice treated with griseofulvin, isogriseofulvin, or 3,5-diethoxycarbonyl-1,4-dihydrocollidine and has been shown to exhibit identical chromatographic characteristics to authentic N-methyl protoporphyrin. All four possible structural isomers have been demonstrated, and each drug produced primarily the same isomer. N-Methyl protoporphyrin has also been found in very small amounts in the liver of untreated mice, but the isomeric composition appeared to differ from that of the drug-induced N-methyl protoporphyrin. Intraperitoneal administration of 3,5-diethoxy-carbonyl-1,4-dihydrocollidine to female C3H/He/Ola and NIH/Ola inbred mice produced a marked dose-related loss of hepatic ferrochelatase activity, which was identical in magnitude in the two strains. Induction of hepatic 5-aminolevulinate synthase (ALA-S), and accumulation of liver protoporphyrin, however, were greater in C3H/He/Ola mice. The strain difference in ALA-S response was most marked when inhibition of ferrochelatase (the "specific" effect of the drug) was maximal, and this suggests that a genetic variation exists in the sensitivity of ALA-S to a second drug action, the so-called nonspecific action, which is shared by many lipid-soluble compounds. Male mice of three strains accumulated greater amounts of hepatic protoporphyrin than females after treatment with griseofulvin, yet no significant difference was found between the two sexes in the extent of ferrochelatase inhibition. Stimulation of ALA-S activity was slightly greater in males, but when porphyria was very marked, ALA-S activities were significantly lower in this sex.(ABSTRACT TRUNCATED AT 250 WORDS)     

Trimethyloxonium modification of single batrachotoxin-activated sodium channels in planar bilayers. Changes in unit conductance and in block by saxitoxin and calcium

Single batrachotoxin-activated sodium channels from rat brain were modified by trimethyloxonium (TMO) after incorporation in planar lipid bilayers. TMO modification eliminated saxitoxin (STX) sensitivity, reduced the single channel conductance by 37%, and reduced calcium block of inward sodium currents. These effects always occurred concomitantly, in an all-or-none fashion. Calcium and STX protected sodium channels from TMO modification with potencies similar to their affinities for block. Calcium inhibited STX binding to rat brain membrane vesicles and relieved toxin block of channels in bilayers, apparently by competing with STX for the toxin binding site. These results suggest that toxins, permeant cations, and blocking cations can interact with a common site on the sodium channel near the extracellular surface. It is likely that permeant cations transiently bind to this superficial site, as the first of several steps in passing inward through the channel.     

NAD metabolism in Salmonella typhimurium: isolation of pyridine analogue supersensitive (pas) and pas suppressor mutants

Mutants of Salmonella typhimurium supersensitive to the nicotinic acid analogue 6-amino-nicotinic acid (6ANA) were isolated as unable to grow on what are normally subinhibitory concentrations of the analogue. The mutations were classified on the basis of their map positions as pasA (89-92 units), pasB (66-69 units), pasC (18-22 units), pasD (18 units) and pasE (55 units). The mutants exhibited a wide range of minimal inhibitory concentrations towards 6ANA, and several were affected in terms of growth. The data suggest that most of the mutations probably reside in genes whose products utilize nicotinamide adenine dinucleotide (NAD) as a cofactor, the altered gene products being more sensitive to internal 6-amino NAD concentrations. Secondary mutations which suppress the Pas- phenotype were found to reside in the following NAD-related loci; pncB, nadB and nadD. Two of the pncB mutants appear to be affected in the expression of NAPRTase while several of the nadB mutants are apparently insensitive to feedback inhibition by internal NAD concentrations.     

Cell shape, spatial patterns of cilia, and mucus-net construction in the ascidian endostyle

The endostyle of the ascidian Ciona intestinalis secretes a mucus-net composed of regularly spaced longitudinal and transverse mucoprotein filaments. It is a gutter-shaped organ composed of eight different longitudinally aligned epithelial zones numbered 1-8. Each zone, with the exception of zone 7 which is unciliatcd. has a characteristic cell shape and spatial pattern of cilia and microvilli. Zones 1.5 and 8 are composed of multiciliated cells, and zones 2,3,4 and 6 of monociliated cells. Cell apices in zones 2, 3 and 4 are rectangular in cross-section, and bear highly ordered rows of cilia. Spacings between cilia both within and between rows are different in each of these zones, but they are similar to interfilament spacings in the mucus-net. The basic structure of the mucus-net is probably secreted and constructed by secretory cells and cilia in zones 1-4. Further secretion may be added in zones 6 and 7 whilst cilia in zones 5, 6 and 8 propel the net out on to the inner surface of the pharynx where it acts as a food filter. The highly organized and structurally complex pattern of ciliated cpithelia in the aseidian endostyle is surprising when compared with the endostyle of the cephalochordate Branchiostoma lanceolatum, which is composed entirely of monociliated cells that differ very little in structure between epithelial zones.     

Relative apparent synapomorphy analysis (RASA). I: The statistical measurement of phylogenetic signal

We have developed a new approach to the measurement of phylogenetic signal in character state matrices called relative apparent synapomorphy analysis (RASA). RASA provides a deterministic, statistical measure of natural cladistic hierarchy (phylogenetic signal) in character state matrices. The method works by determining whether a measure of the rate of increase of cladistic similarity among pairs of taxa as a function of phenetic similarity is greater than a null equiprobable rate of increase. Our investigation of the utility and limitations of RASA using simulated and bacteriophage T7 data sets indicates that the method has numerous advantages over existing measures of signal. A first advantage is computational efficiency. A second advantage is that RASA employs known methods of statistical inference, providing measurable sensitivity and power. The performance of RASA is examined under various conditions of branching evolution as the number of characters, character states per character, and mutations per branch length are varied. RASA appears to provide an unbiased and reliable measure of phylogenetic signal, and the general approach promises to be useful in the development of new techniques that should increase the rigor and reliability of phylogenetic estimates.      

Variation in heat-shock proteins among species of desert fishes (Poeciliidae, Poeciliopsis)

Analysis of the heat-shock proteins (hsps) of six closely related species of Poeciliopsis demonstrated the existence of biochemical diversity in the hsp100, hsp70, hsp60, and hsp30 protein families among species. Each species expressed five to seven hsp70-related isoforms. Constitutive 70-kD isoforms were identical among species, but four different patterns of heat-inducible isoforms were seen in these six species. Members of the hsp70 family of molecular chaperones are included among the most highly conserved proteins known, and the possibility of variation in hsp70 among closely related species has rarely been addressed. The hsp30 family is known to be less conserved than the hsp70 family, and, as expected, the Poeciliopsis hsp30 patterns showed more variation. Most of the hsp30 isoforms characteristic of a particular species were unique to that species. Hsp100 and hsp60 were identical in five of the species, but alternate isoforms were found in P. monacha. The small size and limited geographical distribution of the P. monacha population have probably contributed to the uniqueness of the monacha pattern. Two of the species were shown to acquire thermotolerance, the ability to withstand normally lethal temperatures when subjected to a gradual temperature increase. Rapid-heating protocols commonly used to establish critical thermal maxima of organisms do not include this inducible component of thermoresistance and therefore do not adequately assess an organism's capacity to withstand thermal stress.