Biomedical subjects

A D Levi

Publications and source records attributed to A D Levi.

28 records · Page 2Linked to original sources

PubMed1996

Characterization of the technique involved in isolating Schwann cells from adult human peripheral nerve.

Only recently has it been possible to isolate large quantities of adult derived Schwann cells (SCs) from peripheral nerves in cell culture. These techniques can be easily applied to the isolation of human SCs. We evaluated the influence of donor age and length of explant culture time on the purity of the human SC preparations obtained from a large number (n = 35) of live organ donors ranging in age from 1 to 63 years. The average SC purity from all donors was 92.7 +/- 2.73% and did not appear to be influenced by donor age or duration of culture time in excess of 1 week. Myelin debris was a prominent feature of human SCs prepared in culture and could be detected within histological sections of cultured peripheral nerve segments as well as within human SCs obtained from enzymatic dissociation of the peripheral nerves. This report supports the reproducibility of the techniques involved in isolating human SCs from peripheral nerve from a large series of donors and addresses the mechanism in which a period of cell culture permits the isolation of large quantities of adult human SCs.

Adolescent↗

PubMed1996

Clinical syndromes associated with disproportionate weakness of the upper versus the lower extremities after cervical spinal cord injury.

Patients with cervical spinal cord injuries who present with weakness or paralysis of the hands and arms with relative preservation of lower extremity strengths are often categorized as having two clinical syndromes, cruciate paralysis and acute central cervical spinal cord injury. The explanation for the pathophysiological findings of the dissociated strength in the upper versus the lower extremities has relied on the assumption that there is a localized injury within a somatotopically organized corticospinal tract. This article summarizes the evidence that there is no somatotopic organization within the corticospinal tract in the medulla or cervical spinal cord in primates. An alternative hypothesis for these two syndromes is presented and is based on evidence that has demonstrated that the corticospinal tract in primates is critical for hand function but not for locomotion. Other prevailing theories are reviewed. Thus, we propose that a syndrome consisting of relatively greater hand and arm weakness compared with leg weakness can occur after an injury to the corticospinal tracts in the medulla or the cervical cord. The proposed mechanism, based on the function of the corticospinal tract, unifies a spectrum of injuries of the lower medulla and cervical spinal cord, which produce similar clinical syndromes (cruciate paralysis and acute central cervical spinal cord injury).

Animals↗

PubMed1995

Axon-induced mitogenesis of human Schwann cells involves heregulin and p185erbB2.

The ability of sensory axons to stimulate Schwann cell proliferation by contact was established in the 1970s. Although the mitogen responsible for this proliferation has been localized to the axon surface and biochemically characterized, it has yet to be identified. Recently a family of proteins known as heregulins (HRGs) has been isolated, characterized, and shown to interact with a number of class 1 receptor tyrosine kinases, including the erbB2, erbB3, and erbB4 gene products. These factors include glial growth factor, a Schwann cell mitogen. We have tested the effects of antibodies against components of this system (HRG beta 1 and p185erbB2) in cocultures of rat sensory neurons and human (or rat) Schwann cells to elucidate the role of these proteins in axon-induced Schwann cell proliferation. 2C4, a monoclonal antibody specific for the human p185erbB2 receptor tyrosine kinase, bound to the surface of human Schwann cells and reduced human Schwann cell incorporation of [3H]thymidine by > 90% compared with untreated controls in this coculture system. This antibody had no effect on rat Schwann cell incorporation of [3H]thymidine under similar conditions. A polyclonal antibody raised against HRG beta 1 reduced human and rat Schwann cell incorporation of [3H]thymidine by nearly 80% and up to 49%, respectively, relative to controls. These results imply that a HRG, or a HRG-like molecule, is a component of the axonal mitogen. This mitogen is presented to Schwann cells by axons and induces proliferation through an interaction that involves p185erbB2 on Schwann cells.

Animals↗

PubMed1995

The influence of heregulins on human Schwann cell proliferation.

The use of Schwann cell (SC) autotransplantation to influence neural repair in humans is dependent upon identifying mitogens that will effectively expand human Schwann cells (SCs) in culture. The recent purification and molecular cloning of glial growth factor (GGF), a potent mitogen for rat Schwann cells, has led to the recognition that a family of proteins (GGF/HRG/NDF/ARIA) are alternatively spliced products of a single gene. The heregulins (HRGs) have been characterized with respect to their influence on human breast cancer cell lines; here we examined whether the HRGs have mitogenic activity for human SCs. Using DNA synthesis assays and serial passaging of cells in culture, we demonstrate that HRG is an effective mitogen for human SCs and that, in the presence of agents that elevate cAMP, it is possible to expand these cells over multiple passages without overwhelming fibroblast contamination. One putative target for this family of proteins is p185erbB2, and EGF-like receptor tyrosine kinase that is encoded by the erbB2 protooncogene. In this report we also demonstrate that the erbB2/3/4 messages as well as the erbB2/3 receptor proteins are present within cultured human SCs. The addition of HRG to human SCs results in tyrosine phosphorylation of a 185 kDa protein. In the presence of stimulatory concentrations of HRG, a blocking monoclonal antibody (2C4) to p185erbB2 is capable of significantly inhibiting phosphorylation of a 185 kDa protein as well as the subsequent incorporation of 3H-thymidine within the human SC. These latter results implicate an important role for p185erbB2 in mediating the mitogenic response of human SCs to HRGs.

Adenylyl Cyclases↗

PubMed1994

Cold storage of peripheral nerves: an in vitro assay of cell viability and function.

The development of a nerve bank as a source of donor material to repair large defects in peripheral nerve injuries requires an understanding of the influence of cold storage on cell viability and function in these potential nerve grafts. Segments of peripheral nerves from both human and rat were stored in University of Wisconsin Cold Storage Solution (UW) at 4 degrees C for < 12 h, 3 days, and 1, 2, or 3 weeks. Cellular viability was initially assessed by the degree of cellular outgrowth from explants of the stored nerves placed in culture, and then further quantitated by dissociating the cultured nerve explants and calculating the type and number of cells per milligram of peripheral nerve. Rat Schwann cells (SCs) obtained from the stored (control and 1 and 2 weeks) nerves were tested for their functional ability to myelinate dorsal root ganglion (DRG) neurons in culture. Our findings indicate that human and rat peripheral nerves contain few viable SCs and fibroblasts after 3 weeks of cold storage with the quantity of viable cells within the human cold stored peripheral nerves decreasing significantly after 1 week of cold storage. Despite their reduced number, some SCs from rat nerves stored up to 2 weeks are capable of myelinating DRG axons in culture. These results suggest that short intervals (< 1 week) of cold storage will result in potential peripheral nerve grafts containing large populations of functional cells, while long-term (> or = 3 weeks) cold stored peripheral nerves will contain few viable cells.

Animals↗

PubMed1994

Studies of myelin formation after transplantation of human Schwann cells into the severe combined immunodeficient mouse.

We have previously demonstrated (J. Neurosci., 14: 1309-1319) that Schwann cells (SCs) isolated from adult human peripheral nerve in tissue culture and then transplanted into an immune-deficient rat can enhance axonal regeneration and myelinate regenerating peripheral axons. We have now (a) compared the capacity of both primary and expanded populations of cultured human SCs to form myelin around regenerating mouse axons when transplanted into a gap within the sciatic nerve of severe combined immunodeficiency (scid) mice and (b) also compared the myelinating capability of these cultured SCs to their counterparts in the native human peripheral nerve xenograft. Schwann cells were isolated from adult human peripheral nerve. Semipermeable guidance channels were filled with a 30% Matrigel solution mixed with either primary human SCs or human SCs expanded with mitogens both at a density of 120 million cells/ml. These channels or a human peripheral nerve xenograft were implanted within a 5-mm gap in the transected sciatic nerve of the scid mice and analyzed after a period of 6 weeks. The presence of human myelin segments was confirmed in both the guidance channels containing human SCs and the xenografts by immunostaining with a monoclonal antibody (592) which specifically recognizes a prominent myelin component, P0, in the human but not in the mouse. Within both the guidance channels and the xenografts there was an invasion of the transplant by host SCs which went on to form myelin around regenerating mouse axons. In this report, we also demonstrate that human SCs that have been expanded in culture with mitogens are capable of forming myelin after transplantation in this experimental paradigm.

Animals↗

PubMed1994

The functional characteristics of Schwann cells cultured from human peripheral nerve after transplantation into a gap within the rat sciatic nerve.

The use of human Schwann cells (SCs) in transplantation to promote regeneration in central and peripheral neural tissues must be preceded by efforts to define the factors that regulate their functional expression. Adult-derived human SCs can be isolated and purified in culture, but the culture conditions that allow their full differentiation have not yet been defined. We tested the functional capacity of these cells to enhance axonal regeneration and myelinate regenerating axons in vivo by transplanting them into the damaged PNS of an immune-deficient rat. SCs were purified from human peripheral nerve obtained from organ donors. Semi-permeable guidance channels were filled with a 30% Matrigel containing solution with or without human SCs suspended at a density of 80 x 10(6) cells/ml. Channels were implanted within an 8 mm gap of the transfected sciatic nerve of nude female rats for a period of 4 weeks. Survival of the transplanted human SCs was established by dissociating nerve explants taken from the regenerated cable (after first placing them in culture for 5 d) and staining individual cells for a primate-specific NGF receptor (PNGFr) and S 100. Only one-half of the S 100-positive cells stained for the PNGFr, which indicated that the regenerated cable contained an approximately equal number of human and rat (host) SCs. The presence of some human myelin segments was confirmed by immune staining with an HNK-1 antibody that specifically labels human but not rat myelin. The majority of the myelin segments in the regenerated cable, however, were produced by the rat SCs. The number of myelinated axons and the cross-sectional area of the cable were significantly greater in channels seeded with human SCs when compared to channels containing the diluted Matrigel solution alone. We conclude that purified cultured human SCs can survive and substantially enhance axonal regeneration when transplanted into the injured PNS of an immune-deficient rat. Some of the transplanted human SCs are capable of myelinating regenerating rat axons but are less successful than the host SCs.

Animals↗

PubMed1991

Clival chordoma presenting with acute brain stem hemorrhage.

The authors present a case of a 29-year-old man who developed rapidly progressive cranial nerve palsies and a right hemiparesis secondary to a pontine hemorrhage. The rare but correct diagnosis of a clival chordoma which had invaded the brain stem and subsequently hemorrhaged was based on computed tomography and magnetic resonance imaging. The diagnosis was confirmed at surgery when the patient underwent a successful operative decompression of tumor and clot from the pons via a sub-occipital craniotomy. This represents the first case of a clival chordoma to hemorrhage into the brain stem, which was diagnosed preoperatively and the patient survived.

Acute Disease↗

PubMed1991

Venous thromboembolism after brain tumor surgery: a retrospective review.

We retrospectively reviewed the incidence rate of clinical postoperative deep vein thrombosis and/or pulmonary embolism in 1703 patients undergoing initial craniotomy for meningioma, glioma, or cerebral metastasis. The incidence rate of clinical thromboembolic complications was 1.59% for all tumor groups within the first 4 weeks of surgery. Patients undergoing surgery for meningiomas had a statistically significant increased risk of thromboembolism despite fewer overall perioperative risk factors, when compared with the other tumor groups. The tumor-specific incidence rates of deep vein thrombosis and/or pulmonary embolism for meningioma, glioma, and metastasis were 3.09%, 0.97%, and 1.03%, respectively. Whether this difference was a result of increased surgical time or an inherent property of meningiomas could not be ascertained.

Adult↗

PubMed1990

Spinal hemangioblastoma, syrinx, and hydrocephalus in a two-year-old child.

A two-year-old child presented with an acute inability to bear weight. Radiological investigation revealed a large cervicothoracolumbar syrinx of no known cause. During investigation, acute communicating hydrocephalus developed, which required a shunt. At surgery, a small thoracic spinal cord hemangioblastoma was discovered and excised. Complete recovery with collapse of the syrinx followed. The clinical features of this rare childhood tumor and its associated effects are discussed.

Cerebrospinal Fluid Shunts↗