Ultrasound diagnosis of lymph node metastasis in head and neck cancer.

The role of ultrasonic (US) examination in the detection of cervical lymph node metastasis from head and neck cancer has been evaluated. The subjects were 57 patients with carcinoma of the oral cavity, maxillary sinus or oropharynx who underwent radical neck dissection. The preoperative US and postoperative histopathological findings were compared in 181 lymph nodes (LNs) of 5 mm or more in diameter. LNs were evaluated by US with reference to their size, shape, boundary and internal echoes. The histologically positive rate was higher for larger LNs on US scans: 96% (44/46) of LNs of 15 mm or more were positive. On the other hand, 95% (18/19) of the flat LNs were negative. The positive rate was higher for well delineated than poorly delineated LNs, but similar among the homogeneous, heterogeneous and reflective core patterns of internal echoes. No LNs were detected by US in six of the 57 patients, of which four were true negative and the other two false negative. In the two false negative patients, histopathological examination showed a total of four LNs with two showing extensive extranodal spread of tumour and fibrosis of the surrounding tissue due to previous radiotherapy. Whether LNs are metastatic or not is difficult to determine directly by US. However, the positive rate can be enhanced by evaluation of the size, shape and boundary of the LN. US is indispensable for diagnosing cervical lymph node metastasis in patients with malignant head and neck tumours.

Dopamine-induced depolarizing responses associated with negative slope conductance in LB-cluster neurones of Aplysia.

1. Current- and voltage-clamp methods were used to evaluate the intracellular and ionic mechanisms involved in dopamine-induced slow depolarizations recorded from neurones of the LB cluster in the abdominal ganglion of Aplysia kurodai. 2. In voltage-clamped cells, dopamine induced a slow inward current that, over the range studied (-40 to -110 mV), decreased in amplitude with hyperpolarization of the cell, but failed to invert when the cell was hyperpolarized beyond the reversal potential for K+,(E)K. 3. Bathing the ganglion in 3-isobutyl-1-methylxanthine (IBMX) caused a significant increase in the dopamine response. 4. Most of the responses to dopamine were markedly augmented in Ca2+-free media, but were depressed in Na+-free media. 5. An intracellular injection of cyclic adenosine 3',5'-monophosphate (cyclic AMP) into the same cell type produced an inward current which, like the response to dopamine, diminished in amplitude with hyperpolarization of the cell. 6. Like the dopamine response, the cyclic AMP response increased in the presence of IBMX, was enhanced in Ca2+-free media, was depressed in Na+-free media, and was unaffected by changes in external potassium. 7. In a few cells, although the cyclic AMP-induced responses disappeared in Na+-free media, the dopamine-induced slow inward current responses did not. However, these Na+-free resistant responses disappeared completely in Na+- and Ca2+-free media. 8. It was concluded that most of the dopamine-induced inward current responses were produced by an increase in permeability, mainly to Na+, triggered by a receptor-controlled increase in intracellular cyclic AMP.

Desensitization of Cl(-)-dependent GABA response observed in ganglion cells of Aplysia.

There are many cells in the abdominal ganglion which show a fast, Cl(-)-dependent hyperpolarizing response to gamma-aminobutyric acid (GABA). This response is characterized by an initial rapid increase in membrane conductance followed by an exponential decay to the original value despite a sustained application of GABA. The decay of the response was found to be largely due to the desensitization of the GABA receptor (binding site-ionophore complex) since the equilibrium potential for chloride remained unchanged when the conductance response was depressed. The apparent or measurable rate constant of desensitization (KD) increased when the concentration of GABA increased, showing a saturation in KD-[GABA] relationship at higher concentration of GABA. The rate of desensitization did not change significantly when the resting membrane was hyperpolarized from -40 to -80 mV, though the conductance response was markedly depressed due to a characteristic voltage-dependence in the receptor activation (Matsumoto, 1982). These observations are discussed in terms of an hypothesis in which the desensitization of GABA receptor is the result of an additional binding of a GABA molecule to the activated receptor.

The transmitter-induced change in slope conductance of a receptor membrane as a function of the resting potential and equilibrium potential of the ion involved.

Change in slope conductance has been widely used to evaluate the ion-channel activation by receptor agonists of various postsynaptic membranes. However, the agonist-induced change in slope conductance (delta G) depends not only on the change in membrane ionic permeabilities (delta P) but also on the changes in resting potential (E) and equilibrium potential (EA) of the ion involved (A). A constant field theory was applied to describe delta GA as a function of delta PA, E and EA, assuming that delta PA is not affected by the change in E or EA. This equation predicts that delta GNa will decrease if the membrane is depolarized or when ENa becomes more positive. Similarly, delta GK is expected to increase if the membrane is depolarized or when EK becomes more negative. Further, the equation describes that delta GCl will increase if the membrane is depolarized or when ECl becomes less negative. These changes in GNa, GK and GCl are well consistent with many data previously obtained from various types of receptor membranes. We conclude that the values of slope conductance measured at different E or EA must be carefully corrected in order to estimate the real voltage dependence of delta P or the ion-channel activation by receptor agonists.

Blocking effect of serotonin on inhibitory dopamine receptor activity of Aplysia ganglion cells.

The abdominal ganglion of Aplysia includes neurons with a characteristic dopamine (DA) receptor, the activation of which induces a marked hyperpolarization with a specific increase in the permeability of the membrane to K+. The DA receptor of this type is called the "HK-type." A 2-min exposure to 1 microM serotonin (5-HT) had little effect on the resting membranes with the receptor of HK-type, but significantly depressed the responses to 10 microM DA. The depressing effect of 5-HT on this type of response was completely reversible after a 15-min washing with normal artificial Aplysia blood. Lineweaver-Burke type plotting of the DA-induced responses showed a systematic shift of the straight lines when the concentration of 5-HT was increased; the slope of the line became steeper but the intercept on the ordinate remained unchanged. The dose-inhibition curves, in which relative responses to a given [DA] were plotted against log [5-HT], showed a parallel shift toward the right when the concentration of DA increased. These findings suggest that 5-HT competes with DA for common binding sites at the DA receptor of HK-type, and that the blockade is not due to the interaction of 5-HT with K+-channels in the receptor membrane. The effect of other indole derivatives suggests that the DA receptor of HK-type includes anionic and cationic sites to which the NH2 group and 5-HO group of 5-HT could specifically bind, thus exhibiting competitive blockade.

The fibrillar network in the cytoplasm of squid giant axon extracted with saponin.

Skinned nerve fiber was prepared by treating a giant axon of squid with saponin. Filamentous networks and membranous organelles such as vacuoles, smooth membranes and mitochondria were present after the extraction. Ruthenium red staining of the extracted tissue showed dense fibrillar networks in the cytoplasm that correspond to the "microtrabeculae" or "cytoskeleton" reported by other investigators. Our method of preparation, thus, gives further evidence that these structures are not the result of artifactual condensation or precipitation. The present results also indicated that the networks are composed of acid mucopolysaccharides and/or mucoproteins. The density of microtubules was not changed by the extraction per se, but the number of microtubules remaining after extraction decreased by alteration of environmental conditions such as low temperature (4 degrees C), or the presence of Ca++ (1 mM), Mg++ free medium or colchicine (1 mM). These conditions are known to inhibit axonal transport. The usefulness of our extracted cytoplasm as a model of microtubule-related physiological functions is discussed.

Effects of various enzymes and chemical modification reagents on the Na+- and Cl- -dependent responses of the ganglion cells to acetylcholine.

Using the abdominal ganglion cells of Aplysia, we analyzed the effects of various enzymes and chemical modification reagents on the acetylcholine (ACh)-induced responses of the excitatory (Na+ -dependent) and inhibitory (Cl- -dependent) types. (1) Phospholipase A (2 mg/ml) caused no appreciable effects on either type of response. (2) Phospholipase C (2 mg/ml) markedly depressed both types of response. These suggested that the phosphoryl group of the phospholipid is an important site related to the binding of ACh, common to both types of ACh-receptors. (3) Carboxypeptidase A (10 mg/ml) caused no observable effects on either type of response. (4) Carboxypeptidase B (10 mg/ml) depressed the inhibitory type of response without affecting the excitatory one. (5) Pyridoxal-5'-phosphate (1 mM) also depressed the inhibitory response without affecting the excitatory one. These findings (4, 5) suggested that the Cl- -channel in the inhibitory ACh-receptor complex includes a C-terminal lysine which may play an active role in the movement of Cl- across the receptor membrane. (6) L-Leucine aminopeptidase (1 mg/ml) depressed the excitatory response without altering the inhibitory one. (7) p-Nitrothiophenol (1 mM)-also depressed the excitatory response without affecting the inhibitory one. These findings (6, 7) suggested the presence of a certain N-terminal amino acid near a glutamate or aspartate residue within a molecular moiety of Na+ -channel included in the excitatory ACh-receptor complex.