Coordinated Motion of Epithelial Layers on Curved Surfaces.

Coordinated cellular movements are key processes in tissue morphogenesis. Using a cell-based modeling approach we study the dynamics of epithelial layers lining surfaces with constant and varying curvature. We demonstrate that extrinsic curvature effects can explain the alignment of cell elongation with the principal directions of curvature. Together with specific self-propulsion mechanisms and cell-cell interactions this effect gets enhanced and can explain observed large-scale, persistent, and circumferential rotation on cylindrical surfaces. On toroidal surfaces the resulting curvature coupling is an interplay of intrinsic and extrinsic curvature effects. These findings unveil the role of curvature and postulate its importance for tissue morphogenesis.

Accumulation of PN1 and PN3 sodium channels in painful human neuroma-evidence from immunocytochemistry.

BACKGROUND: The axolemmal distribution and density of voltage-gated sodium channels largely determines the electrical excitability of sprouting neurites. Recent evidence suggests that accumulation of sodium channels at injured axonal tips may be responsible for ectopic axonal hyperexcitability and the resulting abnormal sensory phenomena of pain and paresthesias. For future improvement in pain management it is necessary to identify structurally significant generators of autorhythmicity. A first step in this regard will be to determine the predominant types of sodium channels in injured axons. The opportunity to test human specimens from painful and non-painful neuroma is of great value. METHODS: We employed immunocytochemical methods to investigate if two types of highly specific voltage-gated sodium channel subtypes could be detected in sections of human neuroma. FINDINGS: Both subtypes of sodium channels PN1 and PN3 accumulated abnormally in human neuromas. The immunoreactive pattern was more pronounced in painful neuromas. This is in contrast to previous reports that focused either on PN1 or PN3 as main generators of hyperexcitability induced pain. INTERPRETATION: Both, PN1 and PN3 seem to be involved in hyperexcitability induced pain. It can be expected that a variety of other highly specific voltage gated sodium channel subtypes will be detected in regenerating peripheral nerve in the near future, which contribute to the development of neuropathic pain states. Thus, in order to therapeutically control hyperexcitability induced neuropathic pain, it might be worthwhile to develop pharmaceuticals that can selectively block different sodium channel subtypes and subunits.A review of the role of sodium channels in neuropathic pain is implemented in the discussion.

The effect of vagus nerve stimulation on epileptiform activity recorded from hippocampal depth electrodes.

PURPOSE: To assess the effect of vagus nerve stimulation (VNS) on interictal epileptiform activity in the human hippocampus. Clinical studies have established the efficacy of vagus nerve stimulation in patients with epilepsy (VNS Study Group, 1995), although the electrophysiologic effects of VNS on the human hippocampus and mesial temporal lobe structures remain unknown. METHODS: We report a case study in which a patient with an implanted VNS underwent intracranial electrode recording before temporal lobectomy for intractable complex partial seizures. Epileptiform spikes and sharp waves were recorded from a depth electrode placed in the patient's left hippocampus. Spike frequencies and sharp-wave frequencies before and during VNS were compared using both a 5- and a 30-Hz stimulus. Different stimulation rates were tested on different days, and all analyses were performed using a Student's t test. RESULTS: We found no significant differences in spike frequency between baseline periods and stimulation at 5 and 30 Hz. In contrast, stimulation at 30 Hz produced a significant decrease in the occurrence of epileptiform sharp waves compared with the baseline, whereas stimulation at 5 Hz was associated with a significant increase in the occurrence of epileptiform sharp waves. CONCLUSIONS: VNS produces a measurable electrophysiologic effect on epileptiform activity in the human hippocampus. Although a clinical response to VNS did not occur in our patient before surgery, 30-Hz VNS suppressed interictal epileptiform sharp waves that were similar in appearance to those seen during the patient's actual seizures. In contrast, 5-Hz stimulation appeared to increase the appearance of interictal sharp waves.

Neuronal platelet-activating factor receptor signal transduction involves a pertussis toxin-sensitive G-protein.

In most nonneural systems, platelet-activating factor (PAF) receptor effects are mediated by G-proteins that are often pertussis toxin-sensitive. The activation of pertussis toxin-sensitive G-proteins linked to PAF receptors results in the mobilization of intracellular calcium, at least in part, through the second messenger inositol triphosphate. We have sought to determine if a pertussis toxin-sensitive G-protein is involved in the PAF receptor-mediated phenomena of growth cone collapse and of synaptic enhancement in primary neuronal culture. Using infrared differential interference contrast microscopy and patch-clamp recording techniques, pertussis toxin, but not the inactive B oligomer of the toxin, was found to block both the growth cone collapse and the enhanced synaptic release of excitatory transmitter induced by a nonhydrolyzable PAF receptor agonist, making it likely that Go, Gq, or Gi is the G-protein transducer of PAF receptors in primary neurons. We believe that PAF acts directly on neuronal receptors, which are linked to pertussis toxin-sensitive G-proteins, on the tips of developing neurites, and on presynaptic nerve terminals, leading to growth cone collapse and enhanced synaptic release of transmitter.

The endometrium as a novel target for leptin: differences in fertility and subfertility.

Leptin and its receptor are involved in endocrine and paracrine regulation of metabolism, obesity and reproduction. Here, we describe the detection of the functional long isoform receptor of leptin in human endometrium. The leptin receptor protein was shown to be expressed in glandular and luminal epithelium and is periodically regulated throughout the menstrual cycle, demonstrating main expression in follicular and mid-luteal phase. In contrast, leptin receptor mRNA is detectable by reverse transcription-polymerase chain reaction (RT-PCR) as a constitutive component. Since RT-PCR analyses showed that leptin is not expressed in this tissue, the present study suggests that the human endometrium is a novel target for leptin. Therefore, we investigated 11 subfertile patients who underwent two biopsies in one menstrual cycle. The patients presented with a repetitive endometrial maturation defect, but showed adequate serum hormone concentrations and normal steroid hormone receptor expression and down-regulation in the endometrium. These patients were, however, deficient for expression of the functional leptin receptor. These analyses provide evidence that the lack of the leptin receptor in an ovulatory cycle may contribute to subfertility by a yet undefined 'endometrial factor'.

Abnormal distributions of potassium channels in human neuromas.

Twenty-seven human traumatic neuromas were examined immunocytochemically using highly specific antibodies directed against the voltage-gated delayed-rectifier potassium channel, Kv1.1. Normal sural nerves from six of the above-noted patients served as control specimens. Additionally, nine of the neuromas and two of the sural nerves were immunostained for voltage-gated calcium channels using an antibody that reacts with a wide spectrum of calcium channels. Normal myelinated fibers showed Kv1.1 specific immunoreactivity only at the juxtaparanodal regions. In contrast, within the neuromas approximately 30% of the myelinated fibers exhibited Kv1.1 specific immunoreactivity in dense patches along internodal axonal regions. The clustering of Kv1.1 channels along myelin-ensheathed internodal segments of axon was highly specific for the neuromas, and was never seen in normal nerve. Specific calcium channel immunoreactivity was not detectable in either the neuromas or sural nerves. Taken together with prior studies on sodium channels, these results suggest that selective and specific mechanisms control the distribution of ion channels within neuromas. Further investigation of ion channel changes within neuromas should provide a better understanding of the abnormal axonal hyperexcitability that frequently develops after nerve injury.

Evidence of reinnervation of free patellar tendon autograft used for anterior cruciate ligament reconstruction.

We studied six adult male dogs to determine whether free patellar tendon grafts show evidence of reinnervation when used for anterior cruciate ligament reconstruction. Histologic return of neural elements and return of a somatosensory-evoked potential defined evidence of reinnervation. Before removal, the native anterior cruciate ligament was electrically stimulated with a bipolar electrode and a somatosensory-evoked potential was recorded from a scalp electrode. The ligament was excised and reconstructed using an autogenous patellar tendon graft. Somatosensory-evoked potential was attempted immediately after reconstruction. Histology for nerve endings was performed on the native ligaments. Each animal underwent repeat arthrotomy 6 months later. The grafts were isolated and somatosensory-evoked potentials were attempted. An evoked potential was seen in all six dogs before reconstruction. No graft demonstrated a somatosensory-evoked potential acutely; however, 6 months postoperatively, the somatosensory-evoked potential returned in two cases. Histology of native ligaments showed that 25% of the 100 sections evaluated contained neural elements. Of the receptors present, 89% were mechanoreceptors and 11% were free nerve endings. Histologic examination of the graft tissue 6 months postoperatively revealed that all six grafts also contained neural elements. Mechanoreceptors and free nerve endings were present in approximately equal numbers in the grafts. The results of histology and somatosensory-evoked potential demonstrate that in at least some cases, free patellar tendon grafts show evidence of reinnervation when used for anterior cruciate ligament reconstruction.

Nerve action potential recording method and equipment.

OBJECTIVE: Nerve action potential (NAP) recording is a proven and useful tool in the surgical management of nerve injury. Understanding its materials and methods as well as its pitfalls and limitations is necessary for successful and routine use of this tool. METHODS: The basic electrophysiology of nerve and its implications in the setting of nerve trauma and regeneration are reviewed. Electrode design and electrode placement are discussed. The use of simple electromyographic/nerve conduction machines for intraoperative recording of NAPs is developed, along with some of the technical problems that result from electrophysiological recording in an electrically hostile environment. RESULTS: NAP recording is a good indicator of useful recovery in nerve trauma. The presence and demonstration of an NAP is associated with a 90% chance of recovery to a useful motor state. CONCLUSION: NAP recording provides useful information regarding the regenerative potential of a damaged nerve long before that potential is clinically evident. This information is critical in the optimal management of nerve injuries. The techniques of NAP recording are straightforward, but attention to detail is important, and full understanding of the method is required.

Sodium channel accumulation in humans with painful neuromas.

Painful neuromas from 16 patients were examined using site-specific antisodium channel antibodies employed in immunocytochemical and radioimmunoassay methods. Normal sural nerves from six of these patients served as controls. Immunocytochemistry showed abnormal segmental accumulation of sodium channels within many axons in the neuromas. Dens immunolocalization was especially apparent within the axonal tips. Radioimmunoassay confirmed a significantly greater density of sodium channels in the neuromas as compared with the sural nerves. Thus, sodium channel accumulate abnormally within the axons of neuromas in humans. This alteration of the sodium channels may underlie the generation of axonal hyperexcitability and the resulting abnormal sensory phenomena (pain and paresthesias), which frequently occur after peripheral nerve injury.

[Comparison of the value of angioscopy, angiography and ultrasonography in detection of pathologic vascular processes]. / Gegenüberstellung der Wertigkeit von Angioskopie, Angiographie und Sonographie zur Erfassung pathologischer Gefässwandprozesse.

AIMS: The ability of angioscopy, angiography and sonography to evaluate arteriosclerotic vascular lesions were compared to determine if angioscopy is a valuable addition in the diagnostic armamentarium. METHODS: An in-vitro study was selected to enable microscopic assessment of the arteries. Angiography was followed by sonography and endoscopy and finally the vessels were evaluated macro- and microscopically. RESULTS: The configuration of the vascular stenosis (plaque-shape, semicircular-eccentric, circular-concentric) was more reliably defined with angioscopy as compared to angiography and sonography. Intense calcification of plaque was detected sonographically with a high sensitivity (90%) using the acoustic shadowing and high amplitude criteria. Angioscopy was superior in differentiating between simple and complicated plaques and the assessment of ulcerations, while the visualization of the entire vascular tree with potential collaterals remained the domain of angiography. CONCLUSIONS: Angioscopy seems to be valuable addition in the evaluation of arterial vascular disease. It can be used in the quality control after interventional radiology and surgery.

Penfield Lecture. A quarter century's experience with intraoperative nerve action potential recording.

The rationale, basic considerations, and technique of intraoperative nerve action potential (NAP) recording have been reviewed. Experience using this technique in several thousand patients over a 25 year period has been summarized. The most frequent serious nerve injury is one that leaves it in continuity. Resection of such a lesion that is regenerating does the patient a great disservice as does exploration without repair of one with little or no potential for useful spontaneous recovery. The frequency of lesions in continuity as well as the difficulties in evaluating them in the early months by electromyography let alone by surgical inspection has led to the development of intraoperative NAP recording. Where neurolysis was based on a recordable NAP across a lesion in continuity, 93% recovered good function. Where resection of the lesion was based on absence of an NAP, the injury was, without exception, neurotmetic and/or one with poor potential for useful recovery without repair. Some lesions had an NAP across their lesion but a portion of the cross-sectional area appeared more seriously injured. By use of NAP recordings, a split repair was done and usually with good results.

Enhancement of hippocampal excitatory synaptic transmission by platelet-activating factor.

The biologically active lipid platelet-activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine; PAF) is a mediator of inflammatory and immune responses, and it accumulates in the brain during convulsions or ischemia. We have examined whether PAF may play a second messenger role in the central nervous system by studying effects on synaptic transmission in cultured hippocampal neurons. Carbamyl-PAF, a nonhydrolyzable PAF analog with a similar pharmacologic profile, augmented glutamate-mediated, evoked excitatory synaptic transmission and increased the frequency of spontaneous miniature excitatory synaptic events without increasing their amplitude or altering their time course. This compound had no significant effect on gamma-aminobutyric acid-mediated inhibitory synaptic responses. Lyso-PAF, the biologically inactive metabolic intermediate, had no effect on synaptic transmission. Moreover, the enhancement of excitatory synaptic transmission by carbamyl-PAF was blocked by a PAF receptor antagonist. These results indicate a specific presynaptic effect of PAF in enhancing excitatory synaptic transmission in cultured rat hippocampal neurons.

Intraforaminal repair of plexus spinal nerves by a posterior approach: an experimental study.

Many spinal nerve roots injured due to stretch or other types of lesions are not reparable. Some spinal nerves might be repaired if they could be exposed in their intraforaminal course. A posterior subscapular approach for a more lateral exposure of the brachial plexus was combined with a facetectomy to expose intraforaminal nerves in a series of Macaca rhesus monkeys. This approach exposed a 6- to 10-mm segment of spinal nerve not approachable by a more classic anterior operation. Sural grafts were placed from the dural exit of the spinal nerves to the cord level of the plexus. Nine surviving animals were followed for 36 to 54 months and observed for clinical evidence of return of function. In each animal at least one electromyogram (EMG) was performed. The plexus was then re-exposed and intraoperative nerve action potentials were recorded across graft sites. Evoked muscle action potential and cortical potentials were recorded in six animals. Despite the proximal level of repair, adequate regeneration was shown by clinical, electrical, and histological studies. Functional return was best to the supraspinatus and biceps muscles and to wrist and finger flexors. Clinical recovery was present, but less effective, for deltoid, wrist, and finger extensors and intrinsic muscles of the hand, despite evidence on EMG of reinnervation. Recovery of the infraspinatus muscle was poor. Nerve action potentials could be recorded across each graft site. Reinnervational activity was recorded by EMG and evoked muscle action potential studies in most of the muscles studied, despite the persistence of some denervational changes 3 years or more after injury and repair. Histological studies confirmed the presence of a large number of axons of moderate size and myelination even at the forearm level.

Effect of digitalis on clinical symptoms and conduction variables in patients with multiple sclerosis.

Digitalis has been shown to reverse conduction block in demyelinated nerve fibers in experimental animals. In the search for a symptomatic treatment of multiple sclerosis, digoxin (0.02 mg per kilogram of body weight) was given intravenously to 7 patients with probable or clinically definite multiple sclerosis. All of these patients had temperature-dependent symptoms. In 3 patients, improvement of clinical deficits was observed concurrent with significant changes in evoked potential findings. Digitalis derivatives may be useful in ameliorating symptoms in selected patients with multiple sclerosis.

Joint innervation in joint injury.

When considering joint innervation, it is important to appreciate that the joint receptors and the muscle tendon units about the joint interact in a complimentary manner forming a single afferent system. This system remains complex, and its overall function is still relatively unclear. Although the role of joint afferents can not be explicitly described, it is important to appreciate that alterations in joint innervation caused by athletic trauma can occur and can markedly affect joint function.

Experimental missile wound to the brain.

Among civilians in the United States, 33,000 gunshot wound deaths occur each year; probably half of these involve the head. In combat, head wounds account for approximately half of the immediate mortality when death can be attributed to a single wound. No significant reduction in the neurosurgical mortality associated with these wounds has occurred between World War II and the Vietnam conflict, and very little research into missile wounds of the brain has been undertaken. An experimental model has been developed in the anesthetized cat whereby a ballistic injury to the brain may be painlessly reproduced in order that the pathophysiological effects of brain wounding may be studied and better treatments may be designed to lower the mortality and morbidity rates associated with gunshot wounds. Prominent among physiological effects observed in this model was respiratory arrest even though the missile did not injure the brain stem directly. The incidence of prolonged respiratory arrest increased with increasing missile energy, but arrest was often reversible provided respiratory support was given. It is possible that humans who receive a brain wound die from missile-induced apnea instead of brain damage per se. The mortality rate in humans with brain wounding might be reduced by prompt respiratory support. Brain wounding was associated with persistently increased intracranial pressure and reduced cerebral perfusion pressure not entirely attributable to intracranial bleeding. The magnitude of these derangements appeared to be missile energy-dependent and approached dangerous levels in higher-energy wounds. All wounded cats exhibited postwounding increases in blood glucose concentrations consistent with a generalized stress reaction. A transient rise in hematocrit also occurred immediately after wounding. Both of these phenomena could prove deleterious to optimal brain function after injury.

Hypsarhythmia-infantile spasms in near-drowning: clinical, EEG, CT scan and evoked potential studies.

A 13-month-old child developed hypsarhythmia-infantile spasms after nearly drowning. Serial EEG, multimodality evoked potential, and CT scan of the brain revealed hypsarhythmia, disruption of ascending and descending tracts rostral to the mesencephalon, and a fairly severe degree of dilation of the ventricular system and subarachnoid space. We speculate that deafferentiation of the cerebral cortex is critical in the genesis of the hypsarhythmic pattern.

Neural anatomy of the human anterior cruciate ligament.

The histology of the anterior cruciate ligament was studied by a modified technique of the Gairns gold chloride stain for neural elements. Three morphological types of mechanoreceptors and free nerve-endings were identified: two of the slow-adapting Ruffini type and the third, a rapidly adapting Pacinian corpuscle. Rapidly adapting receptors signal motion and slow-adapting receptors subserve speed and acceleration. Free nerve-endings, which are responsible for pain, were also identified within the ligament. These neural elements comprise 1 per cent of the area of the anterior cruciate ligament.

Delayed paraplegia complicating sublaminar segmental spinal instrumentation.

The cases of two patients with delayed paraplegia after segmental spinal instrumentation with sublaminar wiring are reported. Both patients had complex spinal deformities and had transient neural deficits after the first-stage procedure of anterior release and spine fusion. They had uneventful spinal-cord monitoring during the second-stage procedure of posterior instrumentation and fusion, and function of the lower extremities was present immediately after that operation. Paraplegia then ensued, and was recognized thirty hours later in one patient and six days later in the other. Considering our reproducible and reliable experience (no false-negative results) with spinal cord monitoring in 307 operations, we propose that the delayed onset of paraplegia resulted from a progression of ischemic and edema-producing events that had not developed sufficiently intraoperatively to be reflected by the monitoring. The paraplegia became evident only when the subarachnoid space was obstructed because of progressive postoperative neural edema. The presence of sublaminar implants in narrow, kyphotic segments of the spinal canal probably exacerbated the neural irritation by dural impingement, which was seen myelographically.