A chemically defined medium for the culture of mature oligodendrocytes

A new chemically defined medium consisting of equal parts of Dulbecco modified Eagle's and Ham's F-12 media supplemented with insulin, sodium selenite, putrescine, and D+ galactose, which allows the long-term survival of mature oligodendrocyte pure cultures, is described. Immunohistochemical staining has shown that over 90% of the cells become positive for myelin proteins shortly following subculture. Contaminating astrocytes (2%) do not survive in this medium. Biochemical data have indicated that these purified oligodendrocytes express 2'3'-cyclic nucleotide 3'-phosphohydrolase and UDP-galactose ceramide galactosyltransferase activities. Electron microscopical examination revealed that the oligodendrocytes were mostly of medium-dark type and appeared to be identical to cells cultured in serum-containing medium. The ability to maintain pure oligodendrocyte cultures in such a defined medium will allow investigations concerning exogenous and endogenous factors involved in oligodendrocyte metabolism.     

Use of Vicryl (polyglactin 910) mesh to limit epidural scar formation after laminectomy

A variety of substances have been used at laminectomy sites to prevent postoperative epidural scarring. Free grafts of autologous subcutaneous fat are commonly used both clinically and experimentally. The free fat grafts usually survive, but decrease in size by about 50%. Postoperatively, subcutaneous seroma has been observed with the use of fat grafts, as well as recurrent symptoms of neural compression by the graft that required additional operations. When compared to the use of free grafts after laminectomy in dogs, Vicryl mesh produced slightly more scarring, but consistently less than that observed in control animals. The Vicryl mesh was resorbed by a minimal chronic inflammatory response over about 45 days. Seven of 11 fat-grafted zones showed signs of necrosis, at times with a greater collection of inflammatory cells than that associated with the Vicryl mesh. Of the 4 fat-grafted zones that showed good survival, 2 had gross evidence of neural compression. No surgical zone treated with Vicryl mesh exhibited evidence of neural compression. In view of these results, the use of Vicryl mesh at laminectomy sites may be a safer method of limiting postoperative epidural scar formation.     

Evaluation of metastatic cardiac calcification in a model of chronic primary hyperparathyroidism

Recent reports have fueled an interest in the prevalence and significance of metastatic calcium deposition in patients with chronic hyperparathyroidism. Experimental data are limited by the lack of suitable in vivo animal models. We have developed a model of marked hypercalcemia and overproduction of parathyroid hormone using somatic gene transfer. Briefly, the process involves infection of cultured rodent fibroblasts (RAT-1 cells) with a retroviral expression vector that contains the gene encoding human parathyroid hormone. Fibroblasts are grown to confluence on collagen-coated dextran microcarrier beads and are injected into the peritoneal cavities of syngeneic Fisher rats. Human parathyroid hormone production in rat serum is quantified by an immunoradiometric assay for human parathyroid hormone (1-84), which does not recognize rat parathyroid hormone. These rats consistently show production of human hormone within a week. Levels increase progressively, often to 1 ng/ml within 60 days of injection. Serum calcium showed a concomitant rise to an average of 15.5 mg/dl. In this study, 13 rats that had been transplanted with parathyroid hormone-producing fibroblasts were killed 80 days after injection. Examination of the skeleton revealed demineralization and histopathologic sequelae of parathyroid hormone excess with extensive osteoclastic bone resorption. Examination of the hearts revealed calcification in five of 13 hearts. There was no involvement of major coronary arteries or conducting systems, but there was calcification of cardiac myocytes, primarily in subepicardial region. This model may permit an understanding of the mechanisms for sudden cardiac death in severe hypercalcemia.     

Observations on electric shock treatment

Psychiatric Quarterly -     

Botulinum Toxin A in the Treatment of Chromhidrosis

Background. Chromhidrosis is an uncommon disorder characterized by secretion of colored sweat by apocrine glands, typically localized to the face or axilla. The current treatments available for chromhidrosis are time consuming and frequently ineffective.
Objective. Our purpose is to demonstrate a novel approach to the treatment of apocrine chromhidrosis.
Methods. We report a case of apocrine chromhidrosis successfully treated with botulinum toxin A (BTX-A; Botox).
Results. BTX-A therapy successfully controlled facial chromhidrosis, and the effects were visible at 19 weeks post-treatment. The therapeutic benefits may be attributed to its inhibitory effects on cholinergic stimulation, adrenergic stimulation, and substance P release, although further studies are necessary to elucidate the precise mechanism of action.
Conclusion. This report demonstrates a new therapeutic approach to patients suffering from chromhidrosis.     

Membrane-bound antiviral antibodies as receptors for Sendai virions in receptor-depleted erythrocytes

Anti-Sendai virus antibodies were covalently coupled to neuraminidase-treated human erythrocytes by the use of the bifunctional crosslinking reagents, N-succinimidyl-3-(2-pyridyldithio)propionate or succinimidyl-4-(p-maleimidophenyl)butyrate. Neuraminidase-treated erythrocytes bearing antibodies were able to bind Sendai virus particles, while treated erythrocytes lacking the antibodies failed to do so. Virus particles attached to erythrocyte membranes via the antibodies were able to cause hemolysis (virus-cell fusion) and promoted cell-cell fusion. Similar results were obtained when the antibodies were coupled to cat erythrocytes which lack receptors for Sendai virus particles. Reconstituted Sendai virus envelopes, similar to intact virus particles, were able to hemolyze and to induce fusion of neuraminidase-treated antibody-bearing erythrocytes. However, reconstituted envelopes containing inactive HN (hemagglutinin-neuraminidase) but active F (fusion) glycoproteins, despite attachment to antibody-bearing erythrocytes, failed to hemolyze or to induce cell-to-cell fusion. Fusion could be restored by insertion of an active HN glycoprotein into the membranes of the reconstituted envelopes. These results suggest that the HN glycoprotein, besides being the viral attachment protein, also participates in the membrane fusion process.     

Regulation of experimental autoimmune encephalomyelitis by CD4+, CD25+ and CD8+ T cells: analysis using depleting antibodies

Experimental Autoimmune Encephalomyelitis (EAE) can be induced in mice of the C57BL/6 strain by subcutaneous immunization with myelin/oligodendrocyte glycoprotein (MOG) peptide p35-55 in CFA, administered twice at an interval of one week and supplemented with Bordetella pertussis toxin given IV. Here, we studied the effect on the induction of EAE of depleting antibodies to CD4, CD8, or CD25 administered before either the first or the second dose of MOG p35-55. We found that anti-CD4 abolished EAE when given before the first immunization; anti-CD4 did not affect the disease when it was given before the second immunization. Anti-CD8 enhanced EAE induction when given before either of the two immunizations. Anti-CD25 enhanced EAE to the same degree as anti-CD8 when given before the first immunization, but anti-CD25 was even more effective in enhancing EAE when given before the second immunization. The anti-CD25 treatment led to significantly enhanced IFNgamma production by T cells responding to MOG p35-55 and persisting anti-MOG antibodies detectable 56 days after the first immunization. Administration of anti-CD8 or anti-CD25 abolished the need for pertussis toxin to induce EAE. These findings are compatible with the idea that CD4 T cells are required for the initial induction of EAE and that the disease is down-regulated by T cells expressing CD8 or CD25. These regulatory T cells exist prior to MOG immunization, but the CD25+ regulators appear to be further amplified by immunization.     

Effects of lumbar extensor fatigue and fatigue rate on postural sway

Falls from heights resulting from a loss of balance are a major concern in the occupational setting. Previous studies have documented a deleterious effect of lower extremity fatigue on balance. The purpose of this study was to investigate the effect of lumbar extensor fatigue on balance during quiet standing. Additionally, the effects of fatigue rate on balance and balance recovery rate were assessed. Eight center-of-pressure-based measures of postural sway were collected from 13 participants, both before and after a protocol that fatigued the lumbar extensors to 60% of their unfatigued maximum voluntary exertion force. In addition, postural sway was measured for 30 min after the fatiguing protocol, at 5-min intervals, to quantify balance recovery rate during recovery from fatigue. Two different fatigue rates were achieved by fatiguing participants over either 10 min or 90 min. Results show an increase up to 58% in time-domain postural sway measures with lumbar extensor fatigue, but no change in frequency-domain measures. Fatigue rate did not affect the magnitude of these postural sway increases, nor did it affect the rate of balance recovery following fatigue. Statistical power for the latter result, however, was low. These results show that lumbar extensor fatigue increases postural sway and may contribute to fall-from-height accidents.