AXONAL TRANSPORT OF PHENYLETHANOLAMINE-N-METHYLTRANSFERASE IN TOAD SCIATIC NERVE

—The presence of phenylethanolamine-N-methyltransferase (EC 2.1.1.-) and dopamine-β-hydroxylase (EC 1.14.2.1) activities was demonstrated in the sciatic nerve of the toad, Bufo marinus. The rates of accumulation of phenylethanolamine-N-methyltransferase (PNMT) and dopamine-β-hydroxylase (DBH) proximal to a ligation of the sciatic nerve were studied. DBH accumulated proximal to the ligation at a more than 10-fold faster rate than PNMT. By measuring the rate of loss of enzyme activity distal to a ligation, an estimate of per cent clearance of each enzyme was made. Based on the per cent of enzyme activity free to move, the absolute transport rates for each enzyme were estimated to be: PNMT, 3.6 mm/24 h; DBH, 102 mm/24 h. PNMT activity (89 per cent) was recovered in the soluble fraction of sciatic nerve homogenates with no change occurring in the subcellular distribution of the enzyme proximal to ligations. In contrast, 43 per cent of DBH activity was found in the soluble fraction of sciatic nerve homogenates; but a disproportionate increase in paniculate DBH activity was found proximal to sciatic nerve ligations. Reduction of toad body temperature to 4°C resulted in a complete but totally reversible block of the axonal transport of both PNMT and DBH.     

Levels of myo-inositol in normal and degenerating peripheral nerve

—Free inositol was measured in peripheral nerves of the monkey, rabbit, rat, frog and lobster; levels in mammalian nerve were similar, and two to three times greater than in the other species. Concentrations of myo-inositol in rabbit tibial nerve increased from proximal to distal segments; in optic nerve the concentrations decreased with greater distance from the retina. In the early stages of Wallerian degeneration rabbit tibial nerve contained 25 per cent less free myo-inositol, rat nerve 50 per cent less. Rabbit nerves were analysed at 2 and 5 weeks after section; by 5 weeks levels of myo-inositol had increased to 50 per cent above normal. Similar changes were found in degenerating rabbit optic nerve. The combination of galactose feeding and nerve section resulted in reduction of the myo-inositol in rat sciatic nerve to one-fifth of the control value; galactitol in the nerve decreased by 50 per cent after section. The evidence suggests that myo-inositol in nerve is located mainly in Schwann cells or glia.     

FAST AXONAL TRANSPORT IN VITRO IN THE SCIATIC SYSTEM OF THE FROG

Abstract— An in vitro system from the frog has been used to study fast axonal protein transport. The preparation, which was incubated in a specially made chamber, consisted of the gastrocnemius muscle, the sciatic nerve, the dorsal ganglia and part of the spinal cord. The parts were separated from each other by silicone grease barriers, which made it possible to follow the migration of labelled proteins from the spinal cord and ganglia, along the sciatic nerve, towards the muscle. About 80 per cent of transported proteins in the sciatic nerve originated from the dorsal spinal ganglia and moved antidromically at a rate of 60–90 mm per day at 18°C. The rapidly transported proteins were 90 per cent particulate and mainly associated with structures sedimenting in the microsomal fraction.
The effects of cyclohexirnide showed that the synthesis of rapidly moving proteins and their transport were separate processes. A low concentration of colchicine inhibited the transport when it was present in the medium surrounding the ganglia, but had no effect even at a higher concentration, when it was added to the nerve compartment. The presence of vinblastine at a low concentration in either of the two compartments completely arrested the protein transport. Likewise N-ethylmaleimide or p-chloromercuribenzene sulphonic acid in the nerve medium effectively blocked the fast transport. Results from experiments performed to test the possibility of disto-proximal flow and of transfer of proteins from the muscle to the nerve are discussed.     

THE EXCRETION OF CARBON DIOXIDE BY FROG NERVE

1. Quiescent sciatic nerve of the frog discharges CO₂ at the average rate of 0.00876 mg. CO₂ per gram of nerve per minute. 2. Sciatic nerve steeped one minute in boiling water discharges CO₂ at first at a low rate and after an hour and a half not at all. 3. Degenerated sciatic nerve discharges CO₂ at a slightly higher rate than normal living nerve does. 4. Connective tissue from the frog discharges CO₂ at an average rate of 0.0097 mg. per gram of tissue per minute. 5. Assuming that a nerve is composed of from one-half to one-quarter connective tissue the CO₂ output from its strictly nervous components is estimated to be at a rate of 0.008 mg. CO₂ per gram of nerve per minute. 6. Stimulated sciatic nerve increases the rate of its CO₂ output over quiescent nerve by about 14 per cent. When this number is corrected for strictly nervous tissue the rate is about 16 per cent. 7. The increased rate of CO₂ production noted on stimulation in normal sciatic nerves was not observed when they were boiled, blocked, or degenerated. It was also not observed with stimulated strands of connective tissue.     

DISTRIBUTION OF AMINO ACIDS AND OF EXO- AND ENDOPEPTIDASES ALONG VERTEBRATE AND INVERTEBRATE NERVES

Abstract— The proximo-distal gradients for representative peptidases, peptidylpeptide hydrolases, and amino acids were measured in segments of peripheral nerve from invertebrates and vertebrates and in the lobster brain and ventral cord.
Crustacean nerve was characterized by a large pool of free amino acids totaling 100–200 μmoles/g wet wt. In lobster nerve, the principal free amino acid was aspartic acid which comprised 55 per cent of the free pool, whereas in the rat sciatic nerve it comprised only 5 per cent. The principal free amino acid in rat sciatic nerve was taurine (32 per cent of the pool) and in lobster brain glycine comprised 30 per cent of the pool. No consistent patterns emerged for the gradients along the nerves for amino acids and hydrolytic enzymes. In the leg nerve of the lobster, concentrations of aspartic acid and arginine were higher in the proximal region, and concentrations of proline and alanine were higher in the distal region. Concentrations of most amino acids were higher in the proximal regions of crab nerve, of lobster brain and ventral cord, and of rat sciatic nerve.
Rat sciatic nerve exhibited a pronounced proximo-distal increase in activity of aminopeptidase (Leu-Gly-Gly). In lobster leg nerve, activity of neutral proteinase was higher in the proximal segment, whereas activity of acid proteinase was higher in the distal segment. The best examples of proximo-distal gradients were found in lobster brain and ventral cord; activities of endopeptidases, arylamidases (Leu- and Arg-βNA), and aminopeptidase were higher in the supra-esophageal ganglia or cephalothorax segments than in the distal regions.     

Cavitation in plants at low temperature: is sap transport limited by the tensile strength of water as expected from Briggs' Z-tube experiment?

Xylem cavitation in plants is thought to be caused by a loss of adhesion at the conduit wall surface because a rupture in the body of the water column was implicitly ruled out by an experiment by Lyman J. Briggs with Z-tube capillaries. However, Briggs reported a drastic increase in cavitation pressure of water below 5 degrees C which, if it were also true in xylem conduits, would suggest that water transport in plants could be limited by water cohesion at low temperature. In this study we have repeated Briggs' experiment using stem segments. Xylem vulnerability curves were obtained with a centrifuge technique at 1, 25 and 50 degrees C on yew (Taxus baccata). Contrary to Briggs' finding, vulnerability to cavitation, measured as per cent loss conductance, did not increase sharply at 1 degrees C and was even less than at 25 degrees C and 50 degrees C. Moreover, the onset of cavitation in yew at 1 degrees C was measured at a much more negative pressure than Briggs' value. This points out an artefact in Brigg's experiment at low temperature possibly related to imperfections in the tube walls which act as cavitation nuclei.     

Interposition of a pedicle fat flap significantly improves specificity of reinnervation and motor recovery after repair of transected nerves in adjacency in rats

Despite highest standards in nerve repair, functional recovery following nerve transection still remains unsatisfactory. Nonspecific reinnervation of target organs caused by misdirected axonal growth at the repair site is regarded as one reason for a poor functional outcome. This study was conducted to establish a method for preventing aberrant reinnervation between transected and repaired nerves in adjacency. Rat sciatic nerve was transected and repaired as follows: epineural sutures of the sciatic nerve (group A, n = 6), fascicular repair of tibial and peroneal nerves respectively (group B, n = 8), and, as in group B, separating both nerves using a pedicle fat flap as barrier (group C, n = 8). As control only, the tibial nerve was transected and repaired (group D, n = 5). Muscle contraction force of the gastrocnemius muscle was significantly higher in group C as compared with groups A and B after 4 months. Muscle weight showed significantly lower values in group A as compared with groups B, C, and D. Histologic examination in group C revealed little growth of axons from the tibial to the peroneal nerve and vice versa. This axon crossing was observed only when gaps between the fat cells were available. These findings were confirmed by a significantly lower rate of misdirected axonal growth as compared with groups A and B using sequential retrograde double labeling technique of the soleus motoneuron pool. We conclude that a pedicle fat flap significantly prevents aberrant reinnervation between repaired adjacent nerves resulting in significantly improved motor recovery in rats. Clinically, this is of importance for brachial plexus, sciatic nerve, and facial nerve repair.     

THE ROLE OF WATER IN THE STRUCTURE OF PERIPHERAL NERVE MYELIN

In the study of the drying kinetics of nerve fibres, at least five "phases" of water evaporation can be distinguished. A consideration of the accompanying changes in low-angle x-ray diffraction patterns permits a tentative identification of the "phases" and a quantitative interpretation of the data in terms of the water distribution in nerve fibres. These results suggest that the myelin sheath of frog sciatic nerve contains 40 to 50 per cent water, and it is suggested further that the greater part of this water is "organised" in relation to the hydrophilic groups of the lipide and protein components.     

The effect of heat shock on the growth and mycelial morphology of Streptomyces chrysomallus]

The effect of various conditions of heat shock (1 hour at 35, 38, 40, 42, 45 and 50 degrees C) on the growth and morphological features of Streptomyces chrysomallus, an organism producing actinomycin, was studied. A definite regularity in the mycelium morphological changes at high temperatures was observed. After the shock at 35 and 38 degrees C the biomass volume and morphological features of the streptomycete did not markedly differ from those in the control. The shock at 40 degrees C induced the growth inhibition with decreasing the biomass volume by 50 per cent and appearance of submerged spores. When the shock conditions were more rigid (42, 45 and 50 degrees C) the mycelium growth lacked. It is of interest that the temperature of 42 degrees C induced abundant formation of the spores. With further increasing of the temperature to 45 and 50 degrees C the spore formation was not so abundant. The changes in the growth and development of the streptomycete are discussed in relation to the molecular mechanism of the cell protection from temperature shock.     

A rapid method for the determination of proteolytic activities of enzyme preparations

A method has been described for the determination of proteolytic activities of enzyme preparations using casein as substrate. The rate of digestion is proportional to the enzyme concentration used. This relationship is utilized as a measure of the enzyme activity. One unit of activity is defined as the amount which is required to digest casein in 15 minutes at 37.5 degrees C. so that 50 per cent of the protein in 1 ml. of 0.25 per cent solution is not precipitable by trichloroacetic acid. This method has been used to determine the activity of enzymes from different sources and also used to follow the rate of activation of enzymes.     

Reactions of OH radicals with poly(U) in deoxygenated solutions: sites of OH radical attack and the kinetics of base release

Pulse radiolysis of N2O-saturated solutions of poly(U) in the presence of tetranitromethane showed that 81 per cent of the radicals formed are reducing in nature. Using data from other sources it has been estimated that 70 per cent of the OH radicals add to the base at C(5) and 23 per cent at C(6) while only 7 per cent abstract an H-atom from the sugar moiety. To a large extent the C(5) OH adduct radicals attack the sugar moiety of poly(U) thereby inducing strand breakage and base release. G (base release) = 2.9 can be subdivided into three components: (a) immediate (20 per cent), (b) fast (50 per cent) and (c) slow (30 per cent). The immediate base release must occur either during the free-radical stage or as a result of the rapid (t1/2 less than 4 min at 0 degree C) decomposition of a diamagnetic product. The fast and the slow processes are only readily observable at elevated temperatures, e.g. at 50 degrees C the half lives are 83 min and 26 h, respectively (Ea (fast) = 68 kJ mol-1, Ea (slow) = 89 kJ mol-1, A (fast) = 1.5 X 10(7) s-1, A (slow) = 1.9 X 10(9) s-1. It is concluded that there are three different types of sugar lesions giving rise to base release, structures for which are tentatively proposed.     

Influence of seed moisture content and post-irradiation hydration temperature on the kinetics of reactivity towards oxygen or decay of oxygen-sensitive sites.

The rate of development of post-irradiation oxygen-dependent damage when oxygen is available, and its rate of elimination when seeds are first post-hydrated in oxygen-free water prior to their transfer to oxygenated water, was studied in barley seeds of approximately 3 per cent, approximately 8 per cent and approximately 9 per cent moisture contents at 3 degrees C, 25 degrees C and 37 degrees C. The magnitude of oxic damage at a given dose (35 krad) decreases as the initial seed moisture content increases from approximately 3 per cent to approximately 9 per cent. Significant (P = 0.01) oxic damage is observed in seeds of all the three moisture contents at 3 degrees C and 25 degrees C; however, at 37 degrees C significant (P = 0.01) oxic damage is observed only in seeds of approximately 3 per cent and approximately 8 per cent moisture contents. The magnitude of oxic damage in seeds of a given moisture content remains unaltered following oxygenated post-hydration of seeds at 3 degrees C and 25 degrees C, but it registers a significant (P = 0.01) decrease if post-hydration in oxygenated water is carried out at 37 degrees C. The radiation-induced oxygen-sensitive (An) sites react with oxygen approximately 6 to 8 times faster as compared to their rate of decay in the absence of oxygen at both 3 degrees C and 25 degrees C; however, at 37 degrees C they react only approximately 3 to 4 times faster, in seeds of all the three moisture contents. Moreover, the initiation of the decay of An sites becomes evident much earlier in very dry (approximately 3 per cent moist) seeds than in relatively moist (approximately 8 per cent and approximately 9 per cent) seeds. It is also observed that this fraction of An sites which is capable of a very rapid rate of decay in the absence of oxygen is capable also of an even more rapid rate of reactivity towards oxygen.     

On the soluble phase of adrenergic nerve vesicles: correlation of matrix density and biochemical composition.

Highly purified sympathetic nerve vesicles isolated from bovine splenic nerves were treated by hypo-osmotic shocks, freeze-thawing or incubation in the absence or presence of ATP and MgCl. The vesicle preparations were then studied morphologically by electron microscopy and their content of noradrenaline (NA), and soluble proteins analyzed biochemically with special regard to dopamine beta-hydroxylase (DBH). Hypo-osmotic shocks released about 25 per cent of the NA and protein content and about 8 per cent of the DBH activity. This treatment induced swelling of the vesicles but their membranes remained unruptured and they still contained dense cores. Freeze-thawing released about 35 per cent of the NA, 25 per cent of the proteins and 11 per cent of the DBH. After the latter treatment some matrix material still remained in most vesicles but many were less stainable than the intact vesicles in cold control preparations. During incubation at 30 degrees C in an isotonic sucrose-phosphate medium for 30 min the vesicles released most of their NA and soluble DBH activity as well as much of their matrix density. After incubation at 37 degrees C for 30 min most vesicles appeared translucent. After incubation at 30 degrees C for 30 min in the presence of ATP and MgCl the vesicles lost most of their original NA content but retained their DBH activity and most of their matrix density. The results indicate that there is not always a correlation between NA content and retention of matrix density which suggests that DBH might be a component of a macro-molecular complex responsible for the staining reaction taking place in the maxtrix of NA depleted vesicles. This hypothesis is further supported by the finding of striking similarities between DBH isolated from chromaffin granules and the granular and fibrillar material surrounding the nerve vesicles after depletion.     

Modification of the radiosensitivity of barley seed by post-treatment with caffein. IV. Effect of the moisture content of seed and storage temperature after irradiation.

The oxygen-dependent damage which develops in barley seeds with approximately 7-8 per cent moisture content disappears after post-irradiation storage in vacuo for 48 hours at 40 degrees C and for 24 hours at 50 degrees C. When the diration of storage at 40 degrees C is extended to 384 hours, oxygen-independent damage becomes potentiated. There is oxygen-dependent damage in seeds of approximately 13.3 per cent moisture content and after the seeds have been stored in vacuo at 50 degrees C, the oxygen-dependent damage begins to increase by 168 hours, and it is very significantly potentiated by 192 hours. Under these circumstances, caffeine acts as a radioprotector only as long as the precursors of oxic damage are present in the seeds. Once these sites are lost, caffeine acts only as a radiosensitizer. The oxygen-independent damage which increases with storage at high temperature is further potentiated by caffeine.     

THE USE OF PHOSPHOLIPASE C IN THE STUDY OF RECEPTORS FOR NEUROMUSCULAR BLOCKING AGENTS

Abstract— Segments of diaphragmatic muscle and cerebral nerve-ending particles from the rat were treated exhaustively (180 min) with high concentrations of phospholipase C (300 μg/ml) (EC 3.1.4.3). This enzyme caused loss of approx. 50 per cent of muscle membrane P and 70 per cent of nerve-ending membrane P. At lower concentrations (3 μg/ml; 180 min), phospholipase C released 25 per cent muscle membrane P and 55 per cent of nerve-ending membrane P. At the lower concentration (3 μg/ml; 180 min), the enzyme caused a slow but progressive loss (90 per cent) of muscle twitch amplitude evoked by nerve stimulation (phrenic nerve-hemidiaphragm preparation). Loss of muscle twitch amplitude was notably slowed by removal of the enzyme. Phospholipase C-treated neuromuscular junctions developed no resistance to hemicholinium, botulinum toxin, or d-tubocurarine. Such preparations were unusually sensitive to the neuromuscular blocking action of di-isopropylfluorophosphate. The data do not encourage a belief that phospholipids which are substrates for phospholipase C are receptors for hemicholinium, botulinum toxin, d-tubocurarine or di-isopropylfluorophosphate.     

Non-thermal plasma application enhances the recovery of transected sciatic nerves in rats

This experimental research aimed to investigate the effects of non-thermal plasma on nerve regeneration after transected nerve damage using the sciatic nerve in Wistar albino (A) rats. The experiments were performed on 27 Wistar A rats. The rats underwent surgery for right sciatic nerve exposure and were divided into three groups (each group, n = 9) according to sciatic nerve transected injury (SNTI) and non-thermal plasma application: a non-nerve damage (non-ND) group, a only nerve damage without non-thermal plasma application (ND) group, and a nerve damage with non-thermal plasma application (ND + NTP) group. Subsequent to SNTI and immediate suture, non-thermal plasma was administered three times per week for eight weeks. Evaluation for functional recovery was performed using the static sciatic index measured over the full treatment period of eight weeks. The sciatic nerve specimens were obtained after euthanasia and third day from the last non-thermal plasma application. The sciatic nerve tissues were subjected to histological analysis. Behavior analysis presented that the ND + NTP group showed improved static sciatic index compared with the nerve damage group. Histopathological findings demonstrated that the ND + NTP group had more dense Schwann cells and well-established continuity of nerve fibers, greater than the nerve damage group. Immunohistochemistry showed that the ND + NTP group had increased levels of markers for microtubule-associated protein 2 (MAP2), tau, S100 calcium-binding protein B, and neurofilament-200 and regulated the overexpression of CD68 and MAP2. These results indicated that non-thermal plasma enhanced the motor function and restored the neuronal structure by accelerating myelination and axonal regeneration. Additionally, non-thermal plasma was confirmed to have a positive effect on the recovery of SNTI in rats.     

Walking track analysis: a long-term assessment of peripheral nerve recovery.

Functional recovery following sciatic, tibial, and peroneal nerve injury was assessed over a 1-year period using walking track analysis in the rat. Internal neurolysis did not affect nerve function. Crush injury induced a temporary, but complete, loss of function that recovered to control levels by 4 weeks. Nerve transection resulted in complete loss of function without any evidence of recovery. After nerve repair, functional recovery occurred, reaching near-optimal recovery by 12 weeks. The degree of functional recovery varied with the specific nerve involved. The sciatic nerve recovered 41 percent of function, whereas the tibial nerve recovered 54 percent of function. The peroneal nerve exhibited the highest degree of recovery, achieving functional levels similar to control values. Assessment of neural regeneration using walking track analysis appears to be a valuable addition to the traditional methods of histology and electrophysiology.     

The rate of loss of potassium from human red cells in systems to which lysins have not been added

Curves describing the loss of K from human red cells as a function of time can be interpreted in terms of an equation which treats the K content of the cell (varphi) as the result of an accumulation process occurring at a rate P and an outward diffusion process regulated by a constant a. The equation is useful for describing the observations and for exploring the mechanisms which may be responsible for the K losses, although it cannot be used for analyzing the experimental data in a strict sense in the absence of independent metabolic data because P and a may both be functions of time. The applicability of the equation is illustrated by its use in connection with experimental curves showing K loss as a function of time at 4 degrees , 25 degrees , and 37 degrees C. for systems containing human red cells in isotonic NaCl or NaCl-buffer. At 4 degrees C., the K loss follows an exponential curve approaching an asymptote in the neighborhood of varphi = 0.50 +/- 0.15. The corresponding value of P implies that the cells are able to accumulate about 0.6 per cent of their initial K per hour under these conditions. At 25 degrees C., the K loss starts exponentially but becomes roughly linear with time after 24 to 48 hours. The change of form is probably due to the appearance of autolysins in the system. Curves of a similar mixed or intermediate form may be obtained even at 4 degrees C. if the observations are sufficiently extended and if spontaneous hemolysis becomes appreciable. At 37 degrees C., the K loss is exponential for the first 24 to 36 hours, the curves approaching asymptotes which, translated into terms of P, indicate that the cells can accumulate about 7 +/- 3 per cent of their initial K per hour. After this time autolysis begins to affect the shape of the curves, the rate of K loss increasing rapidly. The effect of adding fluoride or iodoacetate is to lower the position of the asymptote to which the curves proceed; i.e., to decrease the accumulation rate P, to increase the diffusion constant a, or both. Cyanide has almost no effect. Hypotonicity has little effect on the rate of K loss at 37 degrees C.; at 4 degrees C., the rate of loss is somewhat less in hypotonic NaCl. The observation that the K loss in systems at 4 degrees C. and containing as much as 0.086 M NaF does not become complete, but proceeds exponentially towards an asymptote between varphi = 0.2 and 0.4, suggests that 20 to 40 per cent of the cell K is much less diffusible than the remainder at low temperatures and in the absence of lytic substances. A similar conclusion is suggested by the form of the curve for K loss into saline at 4 degrees C., an accumulation rate of 0.6 m. eq./litre of cells/hour at the end of 100 hours or more being improbably great for a system at such a low temperature and containing no added glucose.     

Changes in the sciatic nerve of the rabbit and its tissue constituents during development

Abstract— A segment, defined by gross anatomic boundaries, which comprises most of the sciatic nerve was dissected from rabbits aged from birth to 1 yr. The mean and standard deviation of the variance determined for the weights of pairs of nerve units removed from single animals of an age group did not exceed 2·7 per cent and 12·3 per cent respectively of the average weight of the two nerve units which were the values determined for a group of newborn animals. During the period from birth to one year of age, the weight and length of the nerve unit increased 47- and 6-fold respectively while the animal's weight increased 78-fold. Whereas the nerve unit length increased little after 8 weeks, its weight and that of the whole animal continued to increase up to 1 year. Estimations of nucleic acids in the nerve unit from adult animals gave values of 54 μg DNA-P and 25 μg RNA-P per g fresh weight. During the period birth to 1 yr the absolute amount of DNA in the nerve unit increased 11-fold, 71 per cent of this having occurred by 6 weeks of age. The absolute amount of RNA increased 11 -fold by 8 weeks of age and remained constant thereafter. During the period birth to 1 yr, the proportion of the nerve constituents accounted for by lipid-free solids remained constant at 10 per cent, while lipids increased from 5 to 18 per cent and tissue water declined from 84 to 71 per cent. Changes in some lipid classes were examined also. Increases of absolute content in the nerve unit over the first year and concentrations on a fresh weight basis attained in year-old animals were as follows: cholesterol, 132-fold and 86 mg/g; triglyceride 554-fold and 43·2 mg/g; total phospholipid, 85-fold and 61 mg/g; cerebroside 205-fold and 28 μmol/g; sulphatide, 154-fold and 10 μmol/g; monogalactosyl diglyceride, 31-fold and 0·6 μmol/g. The ratio cholesterol/phospholipid/galactolipid in whole nerve units from adult animals was 2·0/2·1/1·0 and the change in this ratio in the first year was compatible with enrichment of myelin in galactolipid during myelination. The ratio of cerebroside to sulphatide increased from 2·0 to 2·6 during this period. Increases in absolute content and concentrations attained for sialic acid-containing components representing cell surface constituents were as follows: ganglioside sialic acid, 33-fold, and 83 μg/g; glycoprotein sialic acid, 57-fold and 371 μg/g. The quantitative pattern of ganglioside fractions separated by thin-layer chromatography in one direction resembled that of whole brain and was the same at all ages studied. The changes reported reflect a composite picture of changes in a variety of cell membranes indicative of marked variation during development in quantity and composition of membranes.     

The influence of pre-treatment temperature on the thermal sensitivity of a mouse tumour

The response of tumours to hyperthermia was tested by giving graded heat treatments and assessing local control at 90 days. Mice were divided into three groups which were pre-treated for 3 days in ambient temperatures of 4, 21 or 35 degrees C. This enabled the mean tumour resting temperature to be varied by up to 11 degrees C, before subsequent heat treatment. For the heat treatments, the tumours were clamped in order to eliminate blood flow, resulting in uniform temperature distributions and hence more uniform thermal sensitivity. TCD50 values were used to construct Arrhenius plots. For all three pre-treatment temperatures, these plots demonstrated a factor of 1.6 increase in heating time per degree Celsius reduction in heating temperature. However, tumours kept in a 4 degrees C environment before treatment were more thermally sensitive than those kept in 21 degrees C conditions, while those in a 35 degrees C environment were more resistant. Pretreatment at 4 degrees C was equivalent to an increase of either 0.5 degree C in heating temperature or 28 per cent in heating time, compared with pre-treatment at 21 degrees C. Pre-treatment at 35 degrees C was equivalent to a reduction of either 0.6 degree C in heating temperature or 25 per cent in heating time. These data indicate that the pre-treatment tumour temperature is an important parameter, but the effect of heat treatment is more closely related to absolute heating temperature rather than to the increase in temperature above the normal resting level.