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Neoformation of connective tissue occurring at the level of spinal cord injury is considered a factor in the failure of regeneration in the mammalian spinal cord. The purpose of the present research was to experimentally investigate the origin and characteristics of connective proliferation following spinal cord lesion produced by compression in the rat. The role of the dural sheath and that of the tissues surrounding the spinal cord were studied. In one group of animals (1), the dura mater was left intact; in a second group (2) a transverse incision of the dura was performed at the level of the spinal cord compressive lesion. In group (1) a few collagenous fibres were seen within the lesion but no connective septum was observed. In group (2) a transversely orientated septum of fibrous scar tissue was constantly found within the lesioned cord. Our experimental study shows that: 1. dural continuity prevents the formation of connective tissue scarring and limits fibrous reactions in the epidural space; 2. opening of the dural sheath is followed by a vigorous fibroblastic reaction in the epidural tissue which extends into the spinal cord to form a connective septum.

✓ To study oculomotor nerve regeneration in rats, the oculomotor nerve was approached microsurgically and was sectioned at the base of the skull. The nerve stumps were reapproximated and affixed with a plasma clot in Group I animals and were separated by a gap in Group II animals. Visceral eye motility was evaluated weekly between 1 day and 40 weeks after surgery by recording the pupillary diameter under standardized photic stimulation. Somatic eye motility was assessed after 26 weeks by measuring the ocular displacement evoked by vestibular stimulation in the horizontal and vertical planes. Nerve regeneration was documented histologically and morphometrically at 8, 16, and 40 weeks after section. The selectivity of axonal regeneration to the extraocular muscles was investigated after 26 weeks by mapping (with injection of retrograde horseradish peroxidase) the motoneurons that supplied each reinnervated muscle. Between 6 and 20 weeks after section, the pupil diameter showed a progressive reduction in Group I rats, and no changes were observed in Group II rats. Compared with normal rats, the amplitude of horizontal and vertical ocular displacements was decreased, respectively, by 30% and 45% in Group I and by 65% and 80% in Group II. In Group I rats, the vestibular stimulation in the horizontal plane evoked anomalous eye movements with vertical components. On histological examination, regenerated nerves showed a progressive increase of axonal diameter and myelin-sheath thickness. Reinnervated muscles were associated with a less specific, bilateral representation in the midbrain compared with normal muscles, which have unilateral representation. The changes of the somatotopic organization were interpreted as being the result of the misdirected regrowth of axons in the postlesional nerve stump and of the collateral sprouting in the midbrain.

Regenerated central axons have been shown experimentally to penetrate in peripheral nerve segments transplanted into the spinal cord (SC). However, if the nerves are transplanted between the stumps of the transected SC regeneration is impaired by local cavitation and scarring. Our experiment was designed to study whether nerve grafts bridging a severe transverse SC lesion might provide to central regenerating axons a pathway to by-pass the lesion. To this purpose, 2 segments of autologous peripheral nerves were inserted through small dural openings into dorsal longitudinal myelotomies rostral and caudal to a transverse SC lesion in rats. Eighteen weeks after transplantation a large number of well myelinated fibres filled the grafted nerves. Only a few of these fibres, however, could be followed into the SC; they were located in the outer layers of the dorsal white matter. The problems regarding the origin and destination of these fibres are discussed.

✓ Spinal cord tissue was obtained from 13- and 14-day embryonic rats and homologously grafted to the completely transected spinal cord of adult rats. Eight and 12 weeks after grafting, clinical, electrophysiological, histological, and neuroanatomical studies were performed. Motor performance of the hosts was assessed by the inclined-plane test. The conduction of nerve impulses across the lesion-transplantation site was evaluated by recording the spinal corticomotor and somatosensory evoked potentials. The survival, growth, differentiation, and parenchymal integration of the graft were documented histologically on semi-thin sections. The axonal interactions between the host spinal cord and the graft as well as the posttraumatic retrograde degeneration of corticospinal axons were investigated using the horseradish peroxidase (HRP) technique. Clinical and electrophysiological assessments did not demonstrate any functional activity of the graft. On histological examination, grafted neurons showed a survival rate of 55%. Such neurons exhibited a limited degree of growth and differentiation. The extent of parenchymal integration between the host spinal cord and the graft varied considerably among different specimens and in the various regions of every specimen. The HRP investigations demonstrated that some axonal interactions between the host spinal cord and the graft had occurred. Regenerated axons arising from both the spinal cord and the dorsal root ganglia of the host entered the graft and elongated in it. Also, axons from the grafted neurons were able to grow for some distance in the host spinal cord. The phenomenon of the posttraumatic retrograde degeneration of corticospinal axons was not affected by this embryonic tissue grafting.

Total excision of spinal lipomas (SL) is difficult because of the intimate adhesion between the fibro-adipose tissue and the neural structures. Radical surgery in SL can provoke nervous lesions; therefore many authors recommend only partial excision as a means of decompression. Nevertheless the role of traction or tethering of the spinal cord in all spinal disraphism, including SL, is known to be very important in determining the clinical picture. So partial removal of SL cannot be effective in releasing the tethered cord. The use of a CO2 laser at low power (3-5 W) permitted radical surgery without any neural damage and complications in 2 adult patients with large, not well capsulated SL. These patients were previously operated on by the same surgeon with conventional microsurgical techniques for partial excision. After CO2 laser surgery there was significant improvement of the clinical pictures, the follow-up being respectively 24 and 36 months. This clinical experience was performed after a CO2 laser had successfully been used in an experimental model in rats simulating a condition of SL. Using power of 3-5 W was not sufficient to attain an immediate vaporization, but the adipose tissue first changed into a transparent liquid across which it was possible to recognize the interfaces between the neural structures and the adipose tissue.

Cerebrospinal fluid (CSF) Ca2+ and free fatty acids (FFA) levels were determined for 4 h following experimental spinal cord (SC) injury in rabbits. The injury was produced by dropping a 20 g weight from 20 cm on exposed dura mater at the C5-C6 level. Samples of CSF were obtained from the cisterna magna from a second laminectomy at the C1 level. A significant decrease in CSF Ca2+ levels was observed 10 and 30 min post-injury; such levels increased above normal values 180 min post-injury. A significant increase in CSF FFA levels was seen from 10 to 240 min post-injury with a peak at 30 min. The early decrease of CSF Ca2+ levels and the concomitant increase in CSF FFA levels seem to confirm the role of Ca2+ in mediating the lipolytic response of the SC tissue to trauma.

✓ A patient with a long-recognized asymptomatic lipoma adjacent to the deep radial nerve developed paralysis of this nerve from a compression similar to the sleep palsy, or “Saturday-night palsy,” mechanism.

The relationship between the phenomenon of the nonselective reinnervation and the functional recovery after section and repair of the highly organized third cranial nerve motor system in rats was studied. The same relationship after section and repair of the more simply organized sixth cranial nerve motor system in guinea pigs is presented as preliminary results. Anatomical demonstration of nonselective reinnervation was obtained by injecting horseradish peroxidase (HRP) into the extraocular muscles. A bilateral reinnervation of previously ipsilateral innervated muscles both in the third and the sixth nerve was interpreted as a plastic response of the brain stem neurons to the nerve injury. Functional recovery, evaluated by measuring with an infrared light technique the horizontal and vertical vestibulo-ocular reflexes, was excellent for the rectus lateralis muscle while it was relatively poor i.e. partial for the muscles depending on the third nerve. These data suggest that one of the most important factors influencing the functional recovery after section and repair of a peripheral nerve is the complexity of the nerve motor system organization.

✓ The extent of the retrograde degeneration of corticospinal axons following transection of the spinal cord was studied in rats by labeling corticospinal axons with anterogradely transported horseradish peroxidase injected in the sensorimotor cortex. Axotomized corticospinal axons underwent progressive and continuing retrograde degeneration. In specimens examined 5, 14, 28, and 56 days after trauma, the tips of the transected corticospinal axons were seen to terminate at 181 ± 80 µm, 977 ± 203 µm, 1751 ± 344 µm, and 2559 ± 466 µm (mean ± standard deviation), respectively, from the site of transection. The rate of retrograde degeneration varied according to the interval after spinal cord transection, as follows: 36.2 µm/day during the first 5 days; 88.4 µm/day between 5 and 14 days; 55.3 µm/day between 14 and 28 days; and 28.8 µm/day between 28 and 56 days. These findings may serve as useful parameters for the objective assessment of therapeutic modalities in spinal cord injury research.

Autologous sciatic nerve grafts were implanted to the lower thoracic spinal cord (SC) of adult rats. The grafts were longitudinally placed on both sides of the SC midline over the dura mater and their cut ends were inserted into the dorsal white matter of the SC. Eight to 60 weeks later the grafts were exposed. In a first experimental group (A) either horseradish peroxidase (HRP) or lectin conjugated horseradish peroxidase (WGA-HRP) was injected into the grafts in order to investigate the origin and the course of regenerated fibres entering the grafts. In a second experimental group (B) the SC was acutely transected between the upper and lower graft insertions and either HRP or WGA-HRP was injected into the caudal stump of the SC in order to investigate the ability of regenerating axons once entered the grafts to re-enter the SC. In group A retrogradely labelled cells were found in the SC scattered in proximity of both the caudal and rostral graft insertions and in the ventral horns as far as 30 mm rostrally to the grafts. Labelled cells were also located in the spinal ganglia, at the level of the grafts and up to 6 segments caudal to them. In group B retrogradely labelled cells were present in the SC rostrally to the transection, both in proximity of the upper graft insertions and in the ventral horns as far as 30 mm rostrally to the grafts. These findings demonstrate that PN grafts implanted to the SC of adults rats can be innervated by regenerated axons arising from both intraspinal neurons and dorsal root ganglion cells (group A); furthermore axons from intraspinal neurons entered and elongated in the grafts can re-enter the SC (group B).

Following peripheral nerve repair, tension on the suture line is regarded as an important factor in producing scar tissue. In some cases, one or more microsurgical epiperineurium-fascial stitches (EPFS) along the proximal and distal stumps of a transected nerve permit their firm approximation, shifting tensile forces from the suture line over longer segments of the nerve stumps. This simple method was used to treat three ulnar and two cranial nerve lesions with gaps ranging from 8 to 20 mm with an end-to-end suture, avoiding any tension on the suture line. In all cases, functional recovery was satisfactory.

The anatomical location of motoneurons controlling the extraocular muscles within the oculomotor nucleus was investigated by injecting retrograde horseradish peroxidase (HRP) into individual eye muscles in control rats and in rats submitted to intracranial section and repair of the oculomotor nerve 6 months previously. Compared to the controls, the operated animals showed marked changes in the somatotopic organization of the Oculomotor subnuclei. The possible nature of this re-arrangement is discussed.

In adult Wistar rats a segment of the right sciatic nerve was grafted to the right dorsal funiculus of the spinal cord (SC). After survivals of 25–54 weeks a solution of the retrograde fluorescent cytoplasmatic tracer Fast Blue (FB) was injected into the SC and microgranules of the retrograde fluorescent nuclear tracer Diamidino Yellow (DY) were inserted into the graft. DY labelled neurones were found in the SC up to 30 mm both rostrally and caudally to the graft and were always single labelled. DY labelled cells were also found in the dorsal root ganglia (DRG) for 6–7 consecutive levels caudally to the graft. Ten to 30% of the DY labelled DRG cells were double labelled and also contained the FB tracer.

Three pharmacological agents, L-carnitine, L-acetylcarnitine and gangliosides, were tested for their ability to enhance the regeneration of the rat sciatic nerve following transection and microsurgical repair. The drugs were administered intraperitoneally at the dose of 50 mg/kg/d for 28 and 56 d postoperatively. At the end of treatment, the motor function recovery of the peroneal component of the sciatic nerve was assessed and the regenerated nerves were analysed morphometrically on histological semi-thin sections. Also, the reinnervated extensor digitorum longus (EDL) muscles were studied histochemically using the adenosine-triphosphatase (ATP-ase) technique 56 d after surgery. Motor function assessment at 56 d after nerve repair revealed that L-acetylcarnitine-treated animals recovered a clinical grade significantly higher (p less than 0.05) than the control animals. Twenty-eight days after nerve repair, the number of myelinated fibres was significantly higher (p less than 0.05) in L-acetylcarnitine and ganglioside-treated animals than in control animals. However, 56 d after nerve repair the number of regenerated fibres in all the drug-treated groups was not significantly different from that of the control group. The EDL muscles of the drug-treated animals did not show significant differences from those of control animals with respect to fibre composition and fibre diameter although the L-acetylcarnitine-treated animals exhibited a significantly lower (p less than 0.05) degree of muscle atrophy than did the control animals. The results of the present work seem to indicate that L-acetylcarnitine and to a lesser extent gangliosides exert some favourable effect on the regeneration of the transected sciatic nerve in rats.(ABSTRACT TRUNCATED AT 250 WORDS)

The early recognition of incipient Meniere's disease (MD) in the asymptomatic ear of a patient with known unilateral disease has profound implications for patient management and follow-up, but the criteria for a right and precocious diagnosis is still controversial.We evaluated forty-nine patients with MD, selected according to Committee on Hearing and Equilibrium guidelines. All patients underwent laminar tonal audiometry, stapedial reflex study, Glycerol dehydration test, Auditory Brainstem Response (ABR) vestibular examination. MRI was performed in 14 patients.A raised hearing threshold in the contralateral ear was found in 27 patients, but only 7 (14.3%) fulfilled the requirements to be considered affected by bilateral MD. The average delay of occurrence in the contralateral ear was 7 years (from 5 to 12 years). The glycerol test was positive only in 4 patients with unilateral MD and moderate hearing loss. It was not positive in any case of bilateral MD. The membranous endolymphatic duct and sac is not well visualised with MRI on the affected side in the majority of patients.A MRI study must be included in the diagnostic protocol for MD and with improvements in this imaging modality will possibly allow detection of variations in the size of inner ear structures. Glycerol dehydration test was useful only in selected cases. A full assessment of incipient disease in the asymptomatic ear in unilateral Meniere's disease should be undertaken. A conservative approach in surgical treatment of unilateral MD is recommended because of the possibility of evolution of a bilateral form, which can occur even 10 years after the onset of the disease.

Microsatellite instability has been reported in hereditary colorectal cancer syndrome and in various kinds of human sporadic tumors. It has also been shown in brain tumors, although with conflicting results. In the present study, DNA samples obtained from 20 primary brain tumors (10 glioblastomas, three astrocytomas, five meningiomas, one ependymoma, one hemangiopericytoma) were analyzed to detect microsatellite instability. Nine microsatellites, mono, di-, tri- and tetranucleotide repeat markers, located on nine different chromosomes, were used. Four of the 20 neoplasias (20%) showed microsatellite alterations in tumor DNA with respect to normal DNA. Two glioblastomas and one atypical meningioma (15%) showed additional bands or bands with shift of electrophoretic mobility, whereas allelic loss was observed in two glioblastomas (10%). In one glioblastoma, one allelic loss and one extra allele were observed at two different loci. These data indicate that in primary brain tumors there is not a high genetic instability. Although we used markers with inherently high levels of instability, only sporadic microsatellite alterations were found. Consequently, alterations in the mechanisms of DNA mismatch-repair, the most important cause of replication errors in hereditary and sporadic colorectal cancers, do not seem to play a major role in the oncogenesis of primary brain tumors.

The effect of exogenous nerve growth factor (NGF) on axonal regeneration into autologous peripheral nerve (PN) grafts implanted to the spinal cord (SC) of rats was assessed by retrograde labeling of the parent soma of the regenerating axons with horseradish peroxidase. NGF was delivered at the graft site over periods of 15 and 30 days by using indwelling osmotic minipumps. In control rats, the minipumps were filled with saline. At 15 days after grafting in the NGF-treated rats, the mean number of SC as well as dorsal root ganglion (DRG) neurons that regenerated their axons into the peripheral nerve grafts was increased 55.3 and 26.4 times, respectively, as compared to the control group values. At 30 days, SC and DRG neurons in the NGF-treated group were 10.9 and 3.1 times greater than in the control group. In the NGF-treated group, the regenerating SC neurons were located within a range of 7 to 13 mm from the graft site as compared to 1 to 7 mm in the control group. Finally, the analysis of the soma diameters of the regenerating neurons showed that NGF enhanced and maintained with time the regenerative response from small-sized DRG neurons. Therefore, NGF is thought to promote directly the regenerative potential of SC as well as DRG neurons and to exert an indirect glial cell-mediated effect at the SC-graft interface.

Experimental studies on spinal cord (SC) injuries published from 1975 to 1989 in some of the most widely circulating neurosurgical journals were reviewed. The relatively large number of animal species utilized as well as the intensely variable dynamic or static methods employed to induce SC injury represent elements of confusion more than objective necessities in this field of research. In fact, the objective of SC injury research should be to solve the problem of severe SC injuries by either preventing and/or repairing SC damage, rather than looking for modalities to provoke a large spectrum of SC injuries with the result of establishing a correlation between for example, the clinical picture and trauma magnitude. It should be time to study all variables and treatments mainly in only one experimental model. The rat with a permanent paraplegia should represent such a model; the abdominal aorta occlusion for 45 minutes, distal to the renal arteries in rabbits should be the experimental model of choice for ischaemia. If a significant result, such as reversing permanent paraplegia, were obtained in rats, it would be logical to repeat the study in higher mammals and if successful, in humans. For the last decade of this century it is necessary to further study all the mechanisms implied in secondary SC damage as well as to attempt to repair definitive SC damage by using grafts and enhancing the potential regenerative ability of the SC with known and new growth factors. Presently, methylprednisolone, dexametasone, thiopental, naloxone, and hypothermia seem to have some clinical potentials that require studies in humans.

In this study we have examined the effect of propionyl-L-carnitine (PC) on rat spinal cord ischaemia and post-ischaemic reperfusion injury by evaluating two lipid peroxidation indices, thiobarbituric acid reactive substances (TBARS) and diene conjugation, before and after the addition of an ADP-Fe+2 complex to spinal cord homogenates. Aerobic, ischaemic, and post ischaemic reperfusion rat spinal cord homogenates from PC treated and untreated animals did not show any statistically significant difference in their TBARS and conjugated diene content. The addition of the ADP-Fe+2 complex to these homogenates resulted in an increased production of both the lipid peroxidation indices, though the magnitude of such formation was related to the type of experimental intervention. The post-ischaemic reperfusion samples of untreated rats showed the highest TBARS and conjugated diene content, while ischaemic samples in either treated and untreated rats did not show any statistically significant difference with respect to the aerobic samples. The post-ischaemic reperfusion samples of treated rats showed a statistically significant decrease of TBARS and conjugated diene production in comparison to the untreated samples. In addition, PC was also able to partially inhibit TBARS and conjugated diene formation in linoleic acid micelles exposed to hemoglobin, though it did not protect albumin fragmentation from the irradiation of water with an X-ray source.

The recovery cycle of the amplitude of the potential evoked in the cerebral neocortex by paired electrical stimuli of the underlying white matter was studied in 5 epileptic patients with intracerebral electrodes chronically implanted stereotactically. A reproducible pattern was apparent for the late components of the potential (i.e., the peak-to-peak amplitude between the second and the third peak, with an average peak latency of 14 and 35 ms, respectively). There was an early period of facilitation (5-10 ms interstimulus interval) followed by a period of relative or absolute depression (20-100 ms) with recovery at an interstimulus interval of about 150 ms. The recovery function of the early components of the potential (i.e., the peak-to-peak amplitude between the first and the second peak, with an average peak latency of 6 and 14 ms, respectively) was variable; recovery was reached at about 150 ms. The responsiveness seemed less in the most epileptogenic cortical areas.

The anticonvulsant activity of a salt of valproic acid (VA), magnesium valproate (MgV), was assessed against amygdala-kindled seizures in rats. The anti-epileptic power of MgV was compared with that of sodium valproate (NaV). Kindling was obtained by delivering daily to one of the amygdala a 2 s train of monophasic square-wave pulses (1 ms, 60 c.p.s., 100-130 microA) via chronically implanted electrodes. Magnesium valproate and NaV were tested once kindling was stabilized and the post-kindling threshold for generalized convulsions was determined. The drugs were administered intraperitoneally in doses ranging from 25 to 200 mg/kg. The injection/test interval was 30 min. Each animal received a single dose every 24 h. Magnesium valproate exhibited an anticonvulsant activity qualitatively and quantitatively similar to that of NaV. Statistically significant differences were not found between the two drugs with respect to the reduction of seizure severity and afterdischarge (AD) duration. The calculated ED50's were 94.58 and 97.41 mg/kg for the suppression of generalized seizures, 176.96 and 129.26 mg/kg for the suppression of partial seizures, 275.96 and 224.13 mg/kg for the suppression of the local AD in the MgV and NaV treated groups, respectively.

✓ A simple technique is described for protecting the cornea in patients with peripheral facial nerve palsy while waiting for recovery of nerve function. The application of an adhesive strip to the superior eyelid permits opening and closing of the eye, and provides good protection of the cornea.