\4 integrin expression onin vitro andin vivo metastatic variants of lewis lung carcinoma

The expression of 4, 6, and 1 integrin subunits has been investigated on somein vitro andin vivo murine metastatic variants derived from Lewis lung carcinoma (3LL). By the use of monoclonal antibodies which recognizes different epitopes of 6, 1, and 4 subunits we demonstrate that 6 and 1 subunits are expressed in all metastatic variants of 3LL irrespective of their metastatic potential, whereas 4 subunit is expressed only in highly metastasizing cells of 3LL. Northern blots of different metastatic variants probed with 1 and 4 subunits demonstrate thata) significant amounts of 1 mRNA were detected in all metastatic variants of 3LL;b) mRNA corresponding to the described entire coding sequence of 4 subunit is expressed only on highly metastasizing cells of 3LL. We conclude that 4 subunit is specifically expressed in highly metastasizig cells of 3LL while is undetectable in lower metastasizing ones.     

Exocytosis of the prophenoloxidase activating system from crayfish haemocytes

Summary Lipopolysaccharides (LPS) and the -1,3-glucan laminarin G, both of which specifically activate the prophenoloxidase (proPO) activating system of crayfish haemocyte lysate, were found to induce degranulation (exocytosis) and subsequent lysis in vitro of monolayers of semigranular haemocytes from the crayfish,Pacifastacus leniusculus, (Table 1, Fig. 1 b), whereas the granular cells were unaffected (Fig. 1 c).Exocytosis of isolated semigranular or granular cells in vitro could also be evoked by the Ca² ionophore A23187 (Table 2, Fig. 1 d). In this case, the whole proPO system was released from the cellular vesicles in its inactive form, since the secreted material contained protease and prophenoloxidase as inactive proenzymes, which could be activated if LPS or -1,3-glucans were added (Table 3). The anion channel blocker SITS, which inhibits exocytosis in several systems, prevented degranulation triggered by -1,3-glucan, LPS, or ionophore.It is concluded that, in arthropods, LPS serve as an indicator of Gram negative bacteria and -1,3-glucan as an indicator of fungi. These non-self molecules elicit both the exocytotic release of the proPO system from the semigranular cells and the subsequent biochemical activation of this system.Abbreviations CFS crayfish saline - DMSO dimethyl sulfoxide - LDH lactate dehydrogenase - LPS lipopolysaccharide - proPO prophenoloxidase - SITS 4-acetamido-4-isothiocyanato-stilbene-2,2-disulfonic acid, disodium salt     

The effects of pyridine nucleotides on the activity of a calcium-activated nonselective cation channel in the rat insulinoma cell line,CRI-G1

The activity of a calcium-activated nonselective (Ca-NS⁺) channel in a rat insulinoma cell line (CRI-G1) is inhibited by pyridine nucleotides in excised patches. The effects of all four pyridine nucleotides tested, -NAD⁺, -NADH, -NADP⁺ and -NADPH were very similar when tested at 0.1 mm, and at 1 mm the phosphorylated forms, -NADP⁺ and -NADPH, appeared to be slightly more potent than -NAD⁺ and -NADH. All the pyridine nucleotides tested reduced both the open state probability of the channel and the number of functional channels observed in a single patch.The application of -NAD⁺, but not of the other nucleotides tested, to the cytoplasmic surface of isolated inside-out patches from CRI-G1 cells opened a novel nonselective cation channel (the -NAD⁺-NS⁺ channel). The activity of this new channel is calcium sensitive and may also be inhibited by AMP.     

The isolated, perfused head of the marine teleost fish,Myoxocephalus octodecimspinosus: Hemodynamic effects of epinephrine

Summary An isolated, perfused-head preparation utilizing the marine teleost (Myoxocephalus octodecimspinosus) was developed, and the hemodynamic effects of epinephrine on this preparation were studied.The isolated head of the sculpin was found to have a relatively long-term viability as measured by stable perfusion pressures (±4.03 Torr for the first hour of perfusion) and responsiveness to epinephrine for over three hours (Fig. 1). This catecholamine induced a net decrease in the resistance of the vasculature of the head along with an increase in the dorsal aortic blood flow, and a concomitant decrease in a venous flow from the peritoneal cavity and cut muscle mass (Fig. 2, Table 1). Blocking beta adrenergic receptors during epinephrine application produced an increase in the vascular resistance and the dorsal aortic to venous flow ratio (Table 2); beta receptor stimulation was followed by a decrease in the resistance and no change in efferent flow rates (Table 3).It is concluded that alpha adrenergic receptors induce the constriction of the branchial arterio-venous anastomoses and are responsible for the efferent flow rate changes. However, beta adrenergic receptors, presumably at the level of the lamellar arterioles, appear to be the dominant factor in the control of net branchial resistance.     

Eye specialisation in waterstriders: an adaptation to life in a flat world

The angular distribution of the optical axes of the ommatidia in the eyes of waterstriders (Gerris lacustris) has been investigated using antidromic illumination.

Buffering capacity of vertebrate muscle: Correlations with potentials for anaerobic function

Summary Buffering capacities (), measured in slykes (moles of base required to titrate the pH of one gram wet weight of muscle by one pH unit, over the pH range of pH 6 to pH 7) due to non-bicarbonate buffers were measured in locomotory muscles from a variety of terrestrial and marine mammals and teleost fishes (Tables 1 and 2). The highest buffering capacities were found in muscles capable of either intense, burst glycolytic function or prolonged, low-level anaerobic function. Marine mammals had higher muscle buffering capacity on the average than terrestrial mammals. Among the fishes studied, warm-bodied species had the greatest values of all animals examined (Table 2). Deep-sea fishes and shallow-living fishes with sluggish locomotory abilities had low values. Fish white muscle displayed higher buffering capacity than red muscle (Fig. 1; Table 2), in keeping with the more aerobic poise and higher capillary density of the latter type of muscle. Strong correlations were found between (1) and muscle myoglobin concentrations in the mammalian species (Table 1; Fig. 2), and (2) and muscle lactate dehydrogenase (LDH) activities in both the mammals and the fishes (Tables 1 and 2; Fig. 3). No correlation was found between and the activity of a citric acid cycle indicator enzyme, citrate synthase, in the mammalian species. While strongly correlated with buffering capacity, the amounts of myoglobin and LDH in a muscle are not the principal determinants of . The results indicate that muscle intracellular buffering capacity is especially critical in locomotory muscles which must function under conditions (burst locomotion and prolonged, low-level anaerobic function) where circulatory perfusion is inadequate to rapidly remove the acidic end-products such as lactic acid that are produced by anaerobic glycolysis. In this respect, the locomotory muscle of diving mammals and the white skeletal muscles of teleost fishes face a common acid-base regulatory problem and utilize a common biochemical strategy to resolve it.     

Mobility and electric charge of screening pigment granules in the superposition eye ofEphestia kuehniella Z.

Summary The mobility and the electric charge of screening pigment granules of the mealmoth superposition eye were determined electrophoretically in buffer solutions. In potassium phosphate buffer the mobility of the negatively charged granules is linearly dependent on the pH in the range from 4.8 to 7.7 (Fig. 2), and in veronal buffer from pH= 2.3 to pH=7.5 (Fig. 3). At pH=6.6 the values of the effective charge per granule vary between 9.4·10^–17 C and 2.0·10^–16 C, those of the real charge between 2.4·10^–14 C and 5.6·10^–14 C (Table 1, Appendix). For equal electric fields, the mobility of the granules decreases with increasing ionic strength, and it remains the same for > 0.075 mol/1 (Fig. 4).     

Isovelocity investigation of the lengthening behaviour of the erector spinae muscles

The purpose of this study was to investigate the force-velocity (F/) relationship for the erector spinae muscles in submaximal activation movements, with particular attention to their response during lengthening movements and at lower shortening contraction velocities. Dynamic models that predict lower back muscle forces require reasonable representations of the modulating effect of instantaneous velocity. Ten males were observed performing trunk flexion and extension in the sagittal plane under constant load. Contraction velocities were measured as the first derivative from a devise sensitive to changes in spine curvature, and controlled by a visual feedback system while a constant load was applied through a chest harness. The erector spinae exhibited a yielding phenomenon which causes an abrupt drop in force during constant velocity stretching under constant, submaximal, stimulation. The findings were consistent with previous isovelocity muscle lengthening experiments. Yielding appeared dependent on the level of load/activation supporting the theory of a state-variableF/ relationship. The eccentric behaviour of the lower erectors (L3) seemed independent of velocity and length, while that of the upper erectors (T9) showed a dependence on length. At lower concentric velocities, concavity in torque-velocity curves was noted after a threshold velocity. The findings of this study strongly reinforce the notion that theF/ length relationship is not a continuous hyperbolic relationship during muscle shortening and that the commonly modelled force augmentation effect of lengthening is incorrect, at least for submaximal activation of the extensors of the lower back.     

Compartmentation and flux characteristics of ammonium in spruce

Using ¹³NH ₄ ⁺ as a tracer, compartmental analyses for NH ₄ ⁺ were performed in non-mycorrhizal roots of intact Picea glauca (Moench) Voss. seedlings at four different concentration regimes of external NH ₄ ⁺ ([NH ₄ ⁺ ]_o), i.e. 0, 10, 100, and 1500 M. Three kinetically distinct compartments were identified, with half-lives of exchange of approximately 2 s, 30 s, and 14 min, assumed to represent surface adsorption, Donnan free space, and cytoplasm, respectively. No significant differences were found in half-lives of exchange with changes in [NH ₄ ⁺ ]_o. Influx was calculated to be 0.96 mol·g^–1·h^–1 in N-deprived plants (measured at 10 M [NH ₄ ⁺ ]_o), while under steady-state conditions it was 0.21 mol·g^–1h^–1 at 10 M [NH ₄ ⁺ ]_o, 1.96 mol·g^–1h·^–1 at 100 M [NH ₄ ⁺ ]_o, and 6.45 mol·g^–1·h^–1 at 1.5 mM [NH ₄ ⁺ ]_o. Efflux measured over the same range constituted approximately 9% of influx in N-deprived plants, 10% at 10 M, 28% at 100 M, and 35% at 1.5 mM [NH ₄ ⁺ ]_o. Cytoplasmic [NH ₄ ⁺ ] was estimated at 6 m M in N-deprived plants, 2 mM at 10 M [NH ₄ ⁺ ]_o, 14 mM at 100 M, and 33 mM at 1.5 mM. Free-space [NH ₄ ⁺ ] was 84 M, 50 M, 700 M, and 8 mM, respectively. In comparison with previously published data on fluxes and compartmentation of NO ₃ ^– in white-spruce seedlings, results of this study identify a pronounced physiological preference of this species for NH ₄ ⁺ over NO ₃ ^– as an inorganic N source in terms of uptake and intracellular accumulation. The significant ecological importance of this N-source preference is discussed.The research was supported by a Natural Sciences and Engineering Research Council, Canada, grant to Dr. A.D.M. Glass and a University of British Columbia Graduate Fellowship to Herbert J. Kronzucker. Our thanks go to Dr. M. Adam and Mr. P. Culbert at the particle accelerator facility TRIUMF on the University of British Columbia campus for providing ¹³N, to Drs. R.D. Guy and S. Silim for providing plant material, and to Dr. M.Y. Wang, Mr. J. Bailey, Mr. J. Mehroke and Mr. P. Poon for essential assistance in experiments.     

Structure and function of the prehensile tentilla of Euplokamis (Ctenophora,Cydippida)

Euplokamis has coiled tentilla on its tentacles, which can be discharged, flicking out at high velocity, when triggered by contact with prey. The tentillum adheres to prey by means of numerous colloblasts. Discharge, which takes 40–60 ms, is accomplished by contraction of striated muscles, found only in this genus among the Ctenophora. Restoration of the coiled state is attributable to passive, elastic components of the mesogloea. Rows of boxes (fluid-filled compartments) along the sides of the tentillum appear to stiffen the structure so that it does not collapse, kink or buckle during discharge. Smooth muscle fibres present in the tentillum may help pull the tentillum tight after prey have been captured.In addition to the rapid discharge response, the tentillum can perform slower, spontaneous, rhythmic movements which, it is suggested, resemble the wriggling of a plank-tonic worm, enabling the tentillum to function as a lure. These movements appear to be executed by contraction of two sets of myofilament-packed cells which differ in several important respects from conventional smooth muscle. They belong to a novel and distinct cytological subset (inner-ring cells), other members of which are packed with microtubules and seem to be involved in secondary structuring of the collagenous component of the mesogloea.     

Elementary detectors for vertical movement in the visual system of Drosophila

Optomotor thrust responses of the fruitfly Drosophila melanogaster to moving gratings have been analysed in order to determine the arrangement of elementary movement detectors in the hexagonal array of the compound eye. These detectors enable the fly to perceive vertical movement. The results indicate that, under photopic stimulation of a lateral equatorial eye region, the movement specific response originates predominantly from two types of elementary movement detectors which connect neighbouring visual elements in the compound eye. One of the detectors is oriented vertically, the other detector deviates 60° towards the anterior-superior direction (Fig. 5b). The maximum of the thrust differences to antagonistic movement is obtained if the pattern is moving vertically or along a superior/anterior — inferior/posterior direction 30° displaced from the vertical (Fig. 3d,e, Fig. 6). Only one of the detectors coincides with one of the two detectors responsible for horizontal movement detection. This indicates that a third movement specific interaction in the compound eye of Drosophila has to be postulated. — The contrast dependence of the thrust response (Fig. 2) yields the acceptance angle of the receptors mediating the response. The result coincides with the acceptance angle found by analysis of the turning response of Drosophila (Heisenberg and Buchner, 1977). This value corresponds to the acceptance angle expected, on the basis of optical considerations, for the receptor system R 1–6. — The movement-specific neuronal network responsible for thrust control is not homogeneous throughout the visual field of Drosophila. Magnitude and preferred direction of the thrust response in the upper frontal part of the visual field seem to vary considerably in different flies (Fig. 6).     

Tonotopic organisation of the auditory organ of the locustValanga irregularis (Walker)

Summary The responses to pure tone stimuli of individual auditory receptors in the locustValanga irregularis were quantified and the location of receptors within the auditory organ identified through the intracellular injection of Lucifer Yellow. Three groups of receptors were identified and classified according to their site of attachment to the tympanic membrane (Gray 1960). Group a cells were attached to the elevated process and were tuned to sound frequencies from 4 to 6 kHz (Fig. 2). Group c cells were attached to the folded body and were tuned to sound frequencies from 2 to 3.5 kHz (Fig. 3). Group d cells were attached to the pyriform vesicle and were tuned to sound frequencies from 14 to 25 kHz (Fig. 4). The receptor cell bodies belonging to these groups were separated into three distinct regions of the auditory organ (Fig. 5). There was no evidence of any tonotopic organisation within the three groups (Fig. 6). These data therefore support the conclusion that the receptors within the auditory organ of the locust are organized into specific physiological groups (Michelsen 1971a) and thus suggest that the mechanics of the auditory apparatus may indeed play a role in defining the tuning of the receptors.     

Ultrastructural localization of potassium and calcium in an insect ommatidium as demonstrated by X-ray microanalysis

Summary Ultrastructural localization of potassium and calcium in the ommatidium of the house-cricketGryllus domeslicus L. was studied by X-ray microprobe analysis using samples prepared as thin sections (2 or 5 m) of freeze-dried and embedded tissue. Real resolution was limited by the size of ice crystals (Fig. 2) and estimated as about 1 m.Average values for potassium, calcium, sodium and phosphorus in different cells of the compound eye are given in Table 1.Striking non-uniformity in distribution of these elements over the cells and their compartments was found by probe scanning (Figs. 3, 4, 5). The highest potassium and calcium concentrations were measured in the pigmented zones of photoreceptors and pigment cells. The pigment granules are thought to be the ionic depots of the eye.Potassium and sodium are fully accessible to water in sections of embedded tissue, whereas all the calcium and half of the phosphorus are not.The functional significance of the non-uniformity discovered is briefly discussed.     

An optimal monitor of the Rapid-Eye-Movement brain state

A model has been proposed for the stochastic accurrence of bursts of rapid eye movements (REMs) during sleep. REM-bursts are simulated by a Poisson counting process with a rate that depends on a binary Markov sleep state. The corresponding maximum likelihood detector, that continuously monitors the current sleep state based on the observed REM-bursts, has been derived and implemented. Simulations show 97% correct decisions.