Antinuclear antibody with distinct specificity for polymyositis.

In the course of studying antinuclear antibodies in the rheumatic diseases, a new precipitin reaction (provisionally referred to as PM-1) was observed between calf thymus nuclear extract and polymyositis sera. Objectives of this study were to further define the immunologic nature of this reaction and to determine its specificity for polymyositis. Immunodiffusion studies using calf thymus nuclear extract revealed the PM-1 precipitin line in 17 of 28 patients with polymyositis. This reaction was not produced by sera of 460 patients with other diseases. Enzyme and heat treatments of the nuclear extract showed that PM-1 was distinct from native DNA, ribonucleoprotein, and Sm antigens. Fractionation of PM-1-positive serum by 30% ammonium sulphate and Sephadex G-200 chromatography revealed that the factor producing the PM-1 precipitin reaction was in a serum fraction which showed only IgG by immunoelectrphoresis against anti-whole human serum. Because of the apparent strong specificity, the PM-1 system may represent a marker antibody for polymyositis.

Ultrastructure of lymphocyte tumor cell interaction with localization of cell-bound antibody by ferritin labeling.

Splenic lymphocytes derived from Walker carcinoma-bearing rats were harvested and incubated with Walker carcinoma cells growing in tissue culture. The sequence of events leading to target cell death was studied by phase microscopy and scanning and transmission electron microscopy. The sensitized lymphocytes adhere to the tumor cells by multiple cytoplasmic appendages, but no ultrastructural changes are seen at this interface. After 1 hr these lymphocytes release cytoplasmic components consisting of membrane-lined vesicles, cell membranes, endoplasmic reticulum, and cytoplasmic material. This material adheres closely to the surface of the tumor cells and is subsequently seen within the cytoplasm of the tumor cell. The tumor cells then undergo degenerative changes and cell death occurs in 24 to 36 hr. The lymphocyte-derived material appears to contain immunoglobulin components as determined by specific ferritin labeling.

Adaptation of Sinclair swine melanoma cells to long-term growth in vitro.

Sinclair swine melanoma usually regresses in vivo. In the present study, swine melanoma cells were adapted to long-term growth in culture. The morphology of cultured melanoma cells ranged from dendritic to cuboidal, similar to that described for human melanoma cells. Doubling times of the swine melanoma cells were also similar to those of human melanoma cells in vitro. 3,4-Dihydroxy-L-phenylalanine oxidase-positive cells were detected by light microscopy, and melanin and premelanosomes were detected by electron microscopy. Cell cultures could be propagated from progressing, partially regressed, and primary cutaneous lesions, as well as from visceral metastases. Thus, it appears that, under these cell culture conditions, Sinclair swine melanoma cells can be adapted to prolonged growth in vitro.

Immunopathology of skeletal muscle. The value of direct immunofluorescence in the diagnosis of connective tissue disease.

Whitaker and Engel in 1972 first described immunoglobulin deposition in muscle biopsy specimens in connective tissue disorders. In order to confirm and extend their observations and in the hope of identifying features that may differentiate skeletal muscle involvement in connective tissue diseases from other neuromuscular diseases, a series of 80 muscle biopsy specimens were examined using direct immunofluorescence. Three distinctive direct immunofluorescence patterns consisting of vascular, sarcolemma-basement membrane, and fiber staining were identified in skeletal muscle biopsy specimens from 35 patients, 31 of whom had a connective tissue disease. A vascular staining pattern showing a granular deposition of immunoglobulin or complement was seen in 15 patients, all of whom had a connective tissue disease. Routine stains generally did not reveal vessel abnormalities. A sarcolemma-basement membrane staining pattern was demonstrated around the sarcoplasmic membrane in 29 cases. Twenty-six of these patients had a connective tissue disease. There was no correlation with inflammation, fiber necrosis, or degree of connective tissue replacement. Fibers staining for immunoglobulin or complement, seen in 14 cases, generally occurred in morphologically normal fibers. Thirteen of these patients had a connective tissue disease. Since the pathologic change in muscle in the collagen vascular diseases often consists of the nonspecific findings of focal fiber necrosis frequently without inflammatory infiltrates, direct immunofluorescence may be useful in the diagnosis and classification of muscle diseases in the collagen vascular disorders. Furthermore, the findings of immunoglobulin deposition either within vessels or within individual muscle fibers suggest that immunological mechanisms may be responsible for muscular abnormalities in the connective tissue diseases.

The effect of preoperative subconjunctival triamcinolone administration on glaucoma filtration. I. Trabeculectomy following subconjunctival triamcinolone.

Trabeculectomies were performed on 15 eyes because of uncontrolled glaucoma despite the maximum use of tolerated antiglaucoma medication. We considered all of these eyes to be at increased risk for episcleral cicatricial closure for one of the following reasons: neonatal glaucoma, a patient of age 40 years or less, previously failed glaucoma filtration surgery, or aphakia. A standard operation was performed except that triamcinolone acetonide (4 mg) was injected subconjunctivally at the intended trabeculectomy site one week before surgery in 12 eyes, the day of surgery in two eyes, and two days prior to surgery in one eye. During the follow-up period of six to 16 months, 14 of 15 eyes had an intraocular pressure (IOP) of 18 mm Hg or less. The average IOP for these 14 eyes was 12.4 +/- 4.6 (mean +/- 1 SD), and only three of the 14 required any antiglaucoma medication. One eye was a surgical failure. All eyes with controlled IOP had diffuse microcystic filtration blebs by slitlamp examination. The postoperative visual acuity was within one line of the preoperative level in all eyes, except for the one failure. The use of triamcinolone did not appear to add any additional risks to the surgical procedure or to the postoperative period.

Heparin-induced thrombocytopenia, thrombosis, and hemorrhage.

Twenty-two patients developed significant thrombocytopenia (5,000 to 96,000/cu mm; mean, 29,000/cu mm) while receiving prophylactic or therapeutic heparin. Seventeen of them developed serious thrombohemorrhagic complications which accelerated the deaths of six and contributed to the late death of one. Cessation of heparin therapy led to an immediate remission of the thrombohemorrhagic complications and thrombocytopenia, with no patient who was not already moribund dying, once appropriate therapy had been instituted. Platelet-count monitoring is recommended for all patients receiving heparin for more than 6 days, with cessation of heparin therapy mandatory for the successful management of patients with this disorder. Evidence is presented for an immunologic etiology for this disorder.

Atypical fibrous histiocytoma in the skull of an infant. Case report.

This is a report of an atypical fibrous histiocytoma in the skull of an infant, who at the age of 3 months was noted to have a "lump" beneath the scalp in the right parietal region. It was about 2 cm in diameter, and the scalp was movable over it. Physical examination was otherwise normal. Radiographs showed erosion of the scalp deep on the palpable mass. At operation a neoplasm was found, which had destroyed the bone and invaded the adjacent temporal muscle and dura mater. The tumor was removed en bloc after the surrounding bone had been excised. Histological examination using light and electron microscopy revealed the tumor to be an atypical fibrous histiocytoma. Radiation therapy was not given. There has been no recurrence for 7 years.

Interval breast cancers are not biologically distinct--just more difficult to diagnose.

BACKGROUND: Breast cancer diagnosed within 1 year of a negative annual screening examination is called interval breast cancer (IBC) and is considered to be a more virulent subtype of disease. METHODS: We reviewed clinical data on 24 women who were diagnosed as having IBC while participating in the Breast Cancer Detection Demonstration Project at Ellis Fischel Cancer Hospital and the Women's Cancer Control Program screening project in Columbia, Missouri, between 1974 and 1992. We reinterpreted mammograms from the visit prior to the diagnosis of IBC for possible misdiagnosis, changes suggestive of malignancy, and Wolfe's patterns. Archival paraffin blocks from 19 patients were used to determine qualitative expression of tumor markers. RESULTS: Observed 5-, 8-, and 10-year survival rates were identical to published data for patients with non-IBC. Seventy-four percent of the mammograms evidenced dysplastic Wolfe's patterns (P2 and DY), and one patient was found retrospectively to have shown evidence of cancer which was missed. Compared to breast cancers in general, fewer IBC tumors expressed tumor markers associated with poor prognosis. CONCLUSIONS: Survival rates and tumor marker expressions in this retrospective cohort suggest that IBC tumors are not more biologically aggressive than noninterval tumors. They are more difficult to diagnose both by physical examination and mammography.

Origin of muscle action potentials evoked by transcranial magnetic stimulation in cats.

We studied the effects of transcranial magnetic stimulation on ipsilateral and contralateral forelimb extensor muscles in anesthetized cats. A magnetic stimulator, operating at 100% intensity, was used through a circular coil, which was placed tangentially over the midline scalp. Bilateral activation of extensor muscles was readily obtained in all animals. The onset latencies were 7.3 +/- 1.1 and 7.07 +/- 0.8 msec for the contralateral and ipsilateral muscles, respectively. The amplitude of muscle response was unstable in magnitude, nevertheless, it did not show any significant difference between the two sides. The latency of response for ipsilateral and contralateral muscles was similar, which suggests simultaneous activation of motor pathways servicing forelimb muscles. Lesioning or ablation of the motor cortex and decerebration at mid-colliculi level did not abolish the evoked responses elicited at high intensity magnetic stimulation. Stereotactic electrical stimulation of the vestibular nuclei complex was performed, and satisfactory ipsilateral motor responses were obtained. Subsequently, a stereotactic radiofrequency lesion was made at the vestibular nuclei complex, with morphological confirmation. After this lesion, the motor evoked potentials (MEPs) were significantly diminished in amplitude. This finding strongly suggests that the generator of the MEPs resides in the brainstem, mainly at the vestibular nuclei complex.

Lymphocyte-tumor cell interaction in patients with head and neck cancer.

Peripheral lymphocytes from 12 patients with squamous cell carcinoma of the lead and neck were incubated with autologous tumor explants. Four of the 12 patients demonstrated lymphocyte induced tumor cytotoxicity. These lymphocytes adhered to the tumor cells and deposited a radioactive label from their surface onto tumor cells. The deposition of this label was associated with tumor death. Tissue sections from those patients who demonstrated lymphocyte cytotoxicity showed a marked plasmacytic infiltration. This was in contrast to non-responders where only a desmoplastic tissue response was observed with few inflammatory cells.

Structural analysis of monosulfated side-chain oligosaccharides isolated from human tracheobronchial mucous glycoproteins.

To determine the location of some sulfate esters on respiratory mucins, an unambiguous sequencing strategy was developed for a crude, monosulfated oligosaccharide fraction derived from tracheobronchial mucous glycoproteins, isolated from sputum from a patient with cystic fibrosis, and which possessed Ricinus communis-I lectin affinity. Employing fractionation by Bio-Gel P-2 chromatography and high-voltage paper electrophoresis of the pool, eighteen branched and four straight-chained monosulfated oligosaccharides, each possessing at least one neutral D-galactose residue at a nonreducing terminus, were purified. Desulfated analogs of each sulfated oligosaccharide were then produced. Elucidation of their structures and sulfate ester locations was accomplished through a parallel comparative sequencing approach for the sulfated oligosaccharide and its desulfated analog. The method was based on their carbohydrate composition and parallel analysis by sequential exoglycosidase degradations, endoglycosidase digestion, permethylation analyses, and specific lectin affinities. Key to this approach was the inability for specific exoglycosidases and lectins to cleave or bind to, respectively, carbohydrates of their specificity which occupied nonreducing termini and possessed a sulfate ester. Herein we report the structures of twenty-two novel sulfated oligosaccharides. Oligosaccharides ranged from trisaccharides to heptasaccharides, were branched and unbranched, and each possessed a single sulfate ester on either C-6 of a terminal or an internal D-galactose residue or on C-6 of an internal residue of 2-acetamido-2-deoxy-D-glucose (N-acetyl-D-glucosamine).

The clinical significance of the in vivo antinuclear antibody phenomenon.

We describe a patient with systemic lupus erythematosus (SLE) with demonstrated direct immunofluorescent staining of epidermal nuclei in a rim pattern, the second case of in vivo antinuclear antibody (ANA) producing a rim pattern in normal epidermis. A review of the literature showed that 258 cases have been reported of in vivo ANA deposition in epidermal nuclei from nonlesional skin. This phenomenon, which is most common in patients with mixed connective tissue disease, SLE, and scleroderma, correlates frequently with antibodies to extractable nuclear antigens. In vivo ANA have also been described in renal and pulmonary tissue from patients with SLE. In most cases, the pathogenic significance of the phenomenon is unknown.

The effects of taxol, methylprednisolone, and 4-aminopyridine in compressive spinal cord injury: a qualitative experimental study.

BACKGROUND: Taxol is a diterpene alkaloid that stimulates tubulin production in cells. It may be effective in preserving the cytoskeleton of spinal cord axons after injury. METHODS: Thirty-nine rats were submitted to spinal cord compression. The animals were divided into three groups that received taxol (18.75 mg/m2), methylprednisolone (30 mg/kg), or 4-aminopyridine (1 mg/kg). Taxol was administered as one dose immediately after injury and two additional doses on days 14 and 21. Methylprednisolone was given as a single injection immediately postinjury. Four-aminopyridine was administered on days 25, 26, and 27. A group of nine injured animals served as a control without any treatment. Evoked potentials were recorded before, during, and 4 weeks postinjury. Behavioral tests were measured to evaluate recovery of motor function. RESULTS: The taxol and methylprednisolone-treated animals demonstrated a significant improvement in comparison with the control group. No functional improvement was found at 1 mg/kg treatment of 4-aminopyridine in rats. CONCLUSIONS: We conclude that taxol and methylprednisolone given shortly after the compression injury improve functional outcome after an incomplete spinal cord injury.

The inhibition of fibroblast proliferation by a novel monokine: an in vitro and in vivo study.

We have identified a product of rabbit macrophages that inhibits fibroblast proliferation. Tested in vitro against several fibroblast populations, this monokine inhibited rabbit conjunctival fibroblast proliferation by 85% (p = 0.005), human conjunctival fibroblast proliferation by 88% (p = 0.005), human hypertrophic scar fibroblast proliferation by 85% (p = 0.005), and human keloid fibroblast proliferation by 79% (p = 0.005). Additionally (in an in vivo model), this monokine was injected into healing rabbit wounds and inhibited fibroblast proliferation by 33% after 7 days (p = 0.0001) and by 27% after 2 weeks (p < 0.0001). Preliminary analysis of the active factor demonstrates that it is not species-specific, has a molecular weight less than 3000 d, is resistant to degradation by trypsin and carboxypeptidase A, is heat-stable, and is produced by macrophages largely in the first 3 days of culture.

In vitro and in vivo effects of activated macrophage supernatant on distal limb wounds of ponies.

OBJECTIVE: To determine whether monokines produced by activated rabbit peritoneal macrophages can inhibit development of exuberant granulation tissue formation in distal limb wounds in ponies. DESIGN: Randomized block. ANIMALS: 5 castrated male ponies, 2 to 6 years old and weighing 140 to 190 kg. PROCEDURE: In vitro activity of cell-free rabbit peritoneal macrophage supernatant was determined after incubation of fibroblasts from the flank and the distal portion of limbs of horses and ponies. Tritiated thymidine was then added, and after reincubation, radioactivity was measured. After creation of a 4-cm2, full-thickness wound on the mid dorsal aspect of each metacarpus and metatarsus of each pony, in vivo activity of the macrophage supernatant was evaluated. Biopsy specimens were collected at random sites near a border of each wound at 4, 6, and 10 weeks after creation of the wounds. Treatment effects were evaluated on the basis of presence of exuberant granulation tissue requiring excision, number of times that excision was required, total area of the wound, epithelialized area, area of granulation bed, and histologic evaluation of the biopsy specimens. RESULTS: The macrophage supernatant effectively inhibited proliferation of equine fibroblasts in vitro. No significant in vivo treatment effects were found among the 4 treatment groups. CONCLUSION AND CLINICAL RELEVANCE: Monokines from stimulated rabbit peritoneal macrophages may have potential for improving wound healing in horses and ponies because of their effective inhibition in vitro of equine fibroblast proliferation.

Homicide facilitated by inhalation of chloroform.

Three related homicides in which each decedent had significant concentrations of chloroform in blood, fat, brain and/or liver are described. The tissue concentrations of chloroform in one of three decedents were within reported lethal ranges. The concentrations in the remaining two decedents were less than lethal but were well above blood levels in nonoccupationally exposed, healthy subjects. The cause of death in one decedent with sublethal chloroform concentrations was suffocation; the cause of death in the other decedent could not be determined with certainty. The manner of death in each case was homicide. Through a review of the literature the authors discuss the history of chloroform as an inhalation anesthetic and the history of chloroform as an agent of abuse, suicide, assault, and homicide. Blood and/or tissue concentrations of chloroform in nonoccupationally exposed, healthy subjects and victims of suicide or homicide from previous reports are compared and contrasted with the amounts in blood and/or tissue in the three subjects described in this study. The authors conclude that, in addition to a direct lethal effect, chloroform may be used to incapacitate a victim of assault who then dies by another cause.

Inflammatory response to viscoelastic materials.

Healon, Amvisc, or Viscoat was injected into the corneal stroma of 36 normal rabbit eyes. Twelve additional rabbit eyes were used as controls by performing a sham procedure. Twelve rabbits were euthanized at one day and the remainder seven days after injection. The eyes were enucleated and examined by light microscopy. No control eyes had detectable inflammation. Only Viscoat produced mild inflammation at seven days, but this was statistically significant only at p = 0.182. There appears to be little difference in the inflammatory response of these three viscoelastic substances.