Biomedical subjects

T Gordon

Publications and source records attributed to T Gordon.

At least 19 recordsLinked to original sources

PubMed1992

Restricted epitope recognition by precipitin-negative anti-La/SS-B-positive sera.

OBJECTIVE: To determine whether anti-La/SS-B-positive sera that are precipitin negative show a distinct B cell epitope pattern. METHODS: Serum reactivity was tested with recombinant La/SS-B fusion proteins. RESULTS: Among the 18 precipitin-negative anti-La/SS-B-positive sera, reactivity was confined to the full-length recombinant protein (La33.3) in 8 (44%); 5 of 18 (28%) reacted only with La33.3 and with the first 107 N-terminal amino acids (LaA), and 4 (22%) reacted with La33.3, LaA, and the middle region of the La molecule (LaC; amino acids 111-242). One serum reacted with La33.3 and LaC. None of the 18 precipitin-negative sera was positive on a carboxy-terminal fragment (LaL2/3; amino acids 346-408). In contrast, all 26 precipitin-positive anti-La/SS-B-positive sera reacted with La33.3, LaA, and LaC, and 92% reacted with LaL2/3. Rheumatoid factor and serum IgG levels were significantly lower in the precipitin-negative group, providing further evidence of a distinct serologic subset. CONCLUSION: The restricted epitope recognition by these sera may explain the lack of precipitin formation and may represent an early autoantibody response to La/SS-B.

Autoantibodies↗

PubMed1992

Effects of fine and ultrafine sulfuric acid aerosols in guinea pigs: alterations in alveolar macrophage function and intracellular pH.

Acidic sulfate is the most toxicologically important sulfur oxide which exists in the ambient air. To determine if particle size influences toxic effects of sulfuric acid, we investigated the effects of sulfuric acid aerosols of two different sizes on biochemical and cellular parameters of bronchoalveolar lavage fluid from exposed guinea pigs. Guinea pigs were exposed to fine (mass median diameter, 0.3 micron), and ultrafine (mass median diameter, 0.04 micron) sulfuric acid aerosols at 300 micrograms/m3 for 3 hr/day. The animals were euthanized immediately and 24 hr after 1 and 4 days of exposure and lungs were lavaged. Elevated beta-glucuronidase, lactate dehydrogenase activities, and total protein concentration as well as decreased cell viability were observed in the lavage after a single exposure to sulfuric acid aerosols of both sizes. These alterations were small, though statistically significant, and transient. No alteration in these parameters was observed after 4 days of exposure to acid aerosols. In contrast, sulfuric acid-induced alterations in alveolar macrophage function were more pronounced and longer lasting. Immediately after a single exposure to fine acid, there was a 2.7-fold increase in the spontaneous tumor necrosis factor (TNF) release over that in the control group while endotoxin-stimulated TNF release was increased by 2.2-fold. In addition, acid aerosols of both sizes increased the TNF release from macrophages after 4 days of exposure, although there was no clear temporal pattern of induction or recovery. Furthermore, immediately after 4 days of exposure to either fine or ultrafine acid, the amount of H2O2 that could be induced from baseline production by alveolar macrophages was 2.2-fold higher than that of the controls. The phagocytic function of macrophages was also altered by exposure to sulfuric acid aerosols. Twenty-four hours after single or multiple exposure, fine acid enhanced (as high as 78% above control) the in vitro phagocytic activity of alveolar macrophages while ultrafine acid depressed the phagocytic capacity (as much as 50% below that in the control). In addition to these biochemical parameters and cellular functions, we also measured the intracellular pH (pHi) of macrophages harvested after exposures to these acid aerosols using a pH-sensitive fluorescent dye. The resting pHi was depressed after a single exposure to both acid aerosols. The depression in pHi persisted 24 hr after ultrafine acid exposure.(ABSTRACT TRUNCATED AT 400 WORDS)

Aerosols↗

PubMed1992

Rat lung metallothionein and heme oxygenase gene expression following ozone and zinc oxide exposure.

We have conducted exposures in rats to determine pulmonary responses following inhalation of two common components of welding fumes, zinc oxide and ozone. To examine their effects on target-inducible gene expression, we measured mRNA levels of two metal-responsive genes, metallothionein (MT) and heme oxygenase (HO), in lung tissue by RNA slot-blot analysis. A 3-hr exposure to ZnO fume via a combustion furnace caused a substantial elevation in lung MT mRNA at all concentrations tested. Exposures to 5 and 2.5 mg/m3 ZnO resulted in peak 8-fold increases in MT mRNA levels (compared to air-exposed control animal values) immediately after exposure, while 1 mg/m3 ZnO exposure caused a 3.5-fold elevation in MT mRNA. These levels returned to approximate control gene expression values 24 hr after exposure. In addition, ZnO exposure caused an immediate elevation in lung HO gene expression levels, with 8-, 11-, and 5-fold increases observed after the same ZnO exposure levels (p < 0.05). Like MT gene induction, HO mRNA values returned to approximate control levels 24 hr after exposure. In striking contrast to the induction of MT and HO gene expression after ZnO exposures, there was no elevation in gene expression following a 6-hr exposure to 0.5 and 1 ppm ozone, even when lungs were examined as late as 72 hr after exposure. Our results demonstrate the induction of target gene expression following the inhalation of ZnO at concentrations equal to, and below, the current recommended threshold limit value of 5 mg/m3 ZnO. Furthermore, the lack of effect of ozone exposure on MT and HO gene expression suggests no involvement of these genes in the acute respiratory response to this oxidant compound.

Administration, Inhalation↗

PubMed1992

Acute respiratory effects of endotoxin-contaminated machining fluid aerosols in guinea pigs.

Exposure to machining fluid aerosols in the automotive industry is associated with a variety of respiratory symptoms including cross-shift changes in pulmonary function, cough, asthma, and phlegm. Lubricating and cooling fluids used in machining operations are predominantly water and thus are susceptible to microbial growth. In the present study, the role of endotoxin in the acute pulmonary injury produced by machining fluid aerosols was examined in guinea pigs. Animals were exposed to nebulized water, unused machining fluid, or used machining fluid. At the end of a 3-hr exposure, specific airway conductance (SGaw) was not affected by exposure to the vehicle water, but was decreased in a dose-dependent manner by exposure to aerosols of the used machining fluid. SGaw decreased from preexposure baseline values by 0, 7, and 40% in animals exposed to 1, 10, and 100 mg/m3 used machining fluid, respectively. These exposure levels also produced acute lung injury as evidenced by changes in cellular and biochemical indices in lavage fluid. These adverse respiratory effects may have been due to microbial contamination of the used machining fluid as the aerosol exposures were associated with airborne endotoxin concentrations of 0.3, 1.9, and 5.3 micrograms/m3, respectively. Animals exposed to aerosols of the endotoxin-free unused machining fluid had no statistically significant adverse functional, cellular, or biochemical effects except for a fourfold increase in neutrophils at 100 mg/m3. These results suggest that contamination of machining fluid during use or storage may lead to the adverse respiratory effects of aerosolized machining fluids.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗

PubMed1992

Dose-dependent pulmonary effects of inhaled endotoxin in guinea pigs.

As a cell wall component of gram-negative bacteria, endotoxin is thought to play a significant role in the respiratory effects of inhaled organic dusts which are microbially contaminated. Assessment of occupational survey data and clinical studies suggests that few measureable, acute functional changes occur below 30-50 ng/m3 endotoxin (as sampled in airborne dust with a vertical elutriator). Little information is available on the inflammatory effects of inhaled endotoxin at these low concentrations. The present study examined the dose-response relationship between inhaled endotoxin and functional, biochemical, and histological endpoints in the lungs of guinea pigs. Animals were exposed to 0.03 to 50.5 micrograms/m3 aerosolized endotoxin or the vehicle water for 4 hr. At 2 hr into exposure, significant decreases in specific airway conductance were observed only in animals exposed to 9.6 and 50.5 micrograms/m3 endotoxin (17.3 +/- 1.2 and 35.5 +/- 0.5% decreases from baseline values, respectively (mean +/- SE)). Total cell count and lactate dehydrogenase levels in bronchoalveolar lavage fluid were significantly elevated at 24 hr after exposure in all endotoxin-exposed groups except the lowest dose, 0.03 micrograms/m3 (P < 0.05). Polymorphonuclear leukocyte influx into the alveolar region was also dependent on the concentration of inhaled endotoxin. Thus, LDH activity, a biochemical marker of cell injury, and total cell counts and polymorphonuclear leukocytes, markers of inflammation, were more sensitive indices of adverse pulmonary effects from inhaled endotoxin than a functional measurement. These results suggest that subtle inflammatory changes may occur at airborne endotoxin concentrations which may produce no acute respiratory symptoms.

Administration, Inhalation↗

PubMed1992

Airway hyperresponsiveness in guinea pigs exposed to acid-coated ultrafine particles.

Although several epidemiological studies have provided evidence that airborne sulfate particles can produce adverse health effects in susceptible individuals, there is only limited data demonstrating respiratory effects in human volunteers and experimental animals at near ambient concentrations. We have demonstrated previously that the mixing of metal oxide particles with SO2 under humid conditions produces acid-coated particles that are significantly more potent in causing pulmonary function changes than pure acid droplets. The present study examined the nonspecific airway responsiveness to acetylcholine in guinea pigs exposed to acid-coated zinc oxide particles. One and a half hours after a 1-h exposure to the aerosols or a control atmosphere, pulmonary resistance (RL) was measured in awake, spontaneously breathing animals before and during a challenge with increasing doses of iv acetylcholine (Ach). The provocative infusion rate of Ach that resulted in a 100% increase in RL (PR100) was significantly decreased (p less than .05) in animals exposed to sulfuric acid-coated metal oxide particles (approximately 30 micrograms/m3 sulfate) compared to control animals exposed to furnace gases (79.6 +/- 19.4 vs. 179.6 +/- 16.2 micrograms/kg/min, mean +/- SE, respectively). The PR100 of animals exposed to SO2 (109.1 +/- 45.4) or metal oxide particles (106.7 +/- 38.1) alone was not significantly different from that of furnace gas control animals, indicating that the acid coating on the metal oxide particles and not the particles themselves or the SO2 was responsible for the decrease in the PR100. Moreover, a 10-fold greater amount of total sulfate as a pure aqueous sulfuric acid aerosol was necessary to produce a decrease in PR100 (88.6 +/- 11.0 micrograms/kg/min) equivalent to that produced by coated particles. These results suggest that acute exposure to near-ambient concentrations of sulfuric acid under conditions that promote the formation of acid as a surface coating in respirable particles can induce a nonspecific airway hyperresponsiveness. In a similar manner, a dose-dependent significant decrease in PR100 was also produced in animals exposed to sodium sulfite droplets. Thus a single exposure to different forms of sulfur oxide aerosols can induce an alteration in the responsiveness of airway smooth muscle in the guinea pig.

Acetylcholine↗

PubMed1992

Pulmonary effects of inhaled zinc oxide in human subjects, guinea pigs, rats, and rabbits.

Occupational exposure to freshly formed zinc oxide (ZnO) particles (less than 1.0 micron aerodynamic diameter) produces a well-characterized response known as metal fume fever. An 8-hr threshold limit value (TLV) of 5 mg/m3 has been established to prevent adverse health effects because of exposure to ZnO fumes. Because animal toxicity studies have demonstrated pulmonary effects near the current TLV, the present study examined the time course and dose-response of the pulmonary injury produced by inhaled ZnO in guinea pigs, rats, rabbits, and human volunteers. The test animals were exposed to 0, 2.5, or 5.0 mg/m3 ZnO for up to 3 hr and their lungs lavaged. Both the lavage fluid and recovered cells were examined for evidence of inflammation or altered cell function. The lavage fluid from guinea pigs and rats exposed to 5 mg/m3 had significant increases in total cells, lactate dehydrogenase, beta-glucuronidase, and protein content. These changes were greatest 24 hr after exposure. Guinea pig alveolar macrophage function was depressed as evidenced by in vitro phagocytosis of opsonized latex beads. Significant changes in lavage fluid parameters were also observed in guinea pigs and rats exposed to 2.5 mg/m3 ZnO. In contrast, rabbits showed no increase in biochemical or cellular parameters following a 2-hr exposure to 5 mg/m3 ZnO. Differences in total lung burden of ZnO, as determined in additional animals by atomic absorption spectroscopy, appeared to account for the observed differences in species responses. Although the lungs of guinea pigs and rats retained approximately 20% and 12% of the inhaled dose, respectively, rabbits retained only 5%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

The evolution of a case of overlap syndrome with systemic sclerosis, rheumatoid arthritis and systemic lupus erythematosus.

An unusual case of overlap syndrome which evolved over a 12-year period is described. The patient initially presented with limited cutaneous systemic sclerosis. She then developed seropositive erosive rheumatoid arthritis and subsequently vasculitis with positive lupus serology. There was no evidence that she had mixed or undifferentiated connective tissue disease, and antibody to ribonuclearprotein was negative. This unusual combination of connective tissue disorders in one patient is reported and the literature is reviewed.

Arthritis, Rheumatoid↗

PubMed1992

Neural regulation of acetylcholine receptors in rat neonatal muscle.

1. The neuronal regulation of the developmental decline in skeletal muscle acetylcholine (ACh) receptors was studied by comparing the effects of sciatic nerve section or of neuromuscular blockade with botulinum toxin (BoTX) on this decline in neonatal and adult rats, using 125I-alpha-bungarotoxin (125I-BTX) as a ligand for the receptor alpha-subunit. 2. The decline in 125I-BTX binding site concentration in neonatal rat triceps surae muscle homogenates towards low, adult levels followed a simple exponential with a time constant of 8 days. This decline occurred while the muscle is still rapidly growing, before the postnatal increase in numbers of sodium channels. It also preceded the decline in muscle ACh receptor alpha-subunit mRNA, reported in other studies, suggesting that subunit levels are not regulated only by mRNA availability. 3. Muscle denervation in the first two weeks of life prevented this developmental decline. Denervation increased the concentration of 125I-BTX binding sites but the magnitude of this increase became progressively smaller as the muscle matured, showing that removal of innervation during adult life does not revert the muscle, in toto, to its pre-innervation state. 4. Blockade of neuromuscular activity with BoTX increased 125I-BTX binding sites to a lesser extent than muscle denervation during neonatal life. This lesser effect of BoTX blockade contrasts with the equal effects of BoTX blockade and denervation in the adult.

Aging↗

PubMed1992

Laboratory and clinical evaluations of media for the primary isolation of Haemophilus species.

There has not previously been an objective comparison of medium formulations for the primary isolation of Haemophilus species. This study was undertaken to evaluate the components required for the optimal growth of large, easily identifiable colonies of these bacteria. We compared six medium bases and seven supplements for their ability to support the growth of 86 strains of Haemophilus influenzae and 17 strains of other species of Haemophilus. By using a growth index that combines colony size and the dilution factor, a formulation of GC agar base with 1% yeast autolysate and 5% sheep blood (chocolated) promoted the growth of large, easily recognizable colonies of H. influenzae and other Haemophilus species. This medium was designated GCYSB. The addition of hematin to supplements that supplied NAD (or factor V) to the medium was inhibitory to the growth of all of the Haemophilus species tested. In a clinical comparison of GCYSB with routinely used chocolate agar medium in two laboratories for the primary isolation of Haemophilus species, overall GCYSB promoted better growth of 124 strains of H. influenzae and H. parainfluenzae. GCYSB is easy to prepare and inexpensive compared with the ease of preparation and expense of other Haemophilus isolation media.

Bacteriological Techniques↗

PubMed1992

Recovery of rat tibialis anterior motor unit properties following partial denervation.

The recovery of selected mechanical, morphological, and metabolic properties of rat tibialis anterior fast motor units was determined following partial denervation (n = 7) or partial denervation and hemispinal cord transection (n = 5) and compared with age-matched control units (n = 7). Following 1-12 months of recovery, the mechanical properties of each unit were measured and the fibres depleted of glycogen by using standard ventral root filament stimulation techniques. Quantitative histochemical techniques were used to determine cross-sectional area and the activities of succinate dehydrogenase and alpha-glycerolphosphate dehydrogenase in individual unit fibres. Partial denervation increased the mean fibre area but decreased alpha-glycerolphosphate dehydrogenase activity. Succinate dehydrogenase was unchanged in the denervated groups. The variability in area and enzymatic activities among the unit fibres was unchanged. However, the interrelationship between the enzymes was altered by both denervation procedures. Succinate dehydrogenase activity was directly related to fatigue resistance and inversely related to tetanic tension across the units. These findings suggest that a motor unit reestablishes many of its properties despite marked changes to the composition of the unit brought about by partial denervation. In addition, a reduction in the neuromuscular activity of units during reorganization had a limited effect on recovery.

Animals↗

PubMed1992

Optimal stimulation of paralyzed muscle after human spinal cord injury.

Muscle properties change profoundly as a result of disuse after spinal cord injury. To study the extent to which these changes can be reversed by electrical stimulation, tibialis anterior muscles in complete spinal cord-injured subjects were stimulated for progressively longer times (15 min, 45 min, 2 h, and 8 h/day) in 6-wk intervals. An index of muscle endurance to repetitive stimulation doubled (from 0.4 to 0.8), contraction and half-relaxation times increased markedly (from 70 to approximately 100 ms), but little or no change was measured in twitch or tetanic tension with increasing amounts of stimulation. The changes observed with 2 h/day of stimulation brought the physiological values close to those for normal (control) subjects. A decrease in the stimulation period produced a reversal of the changes. No effects were observed in the contralateral (unstimulated) muscle at any time, nor was there evidence of decreased numbers of motor units in these subjects secondary to spinal cord injury. Motor unit properties changed in parallel with those of the whole muscle. The occasional spasms occurring in these subjects are not sufficient to maintain normal muscle properties, but these properties can largely be restored by 1-2 h/day of electrical stimulation.

Adaptation, Physiological↗

PubMed1992

Innervation ratio is an important determinant of force in normal and reinnervated rat tibialis anterior muscles.

1. The technique of glycogen depletion and periodic acid-Schiff (PAS) staining, which identifies glycogen-free muscle fibers, was used to directly count the number (N) and measure the cross-sectional area (CSA) of muscle fibers in single motor units (MUs) from normal and reinnervated tibialis anterior (TA) muscles. Indirect estimates, derived from the proportions of muscle fiber types to MU types, were also made, and force per unit area (or specific force, SF) was calculated. Previous results using direct and indirect approaches have been contradictory. To shed more light on this issue, the relative contributions of N, mean fiber area (A), and SF to muscle-unit force were determined by the use of both methods. 2. TA muscles were examined in experimental rats 3.5-10 mo after cutting and resuturing the common peroneal nerve in one hindlimb and in muscles in age-matched control rats. Ventral roots were dissected to isolate and characterize single MUs according to contraction speed, sag, and fatigability. One unit per muscle was selected for repetitive tetanic stimulation designed to deplete muscle fiber glycogen stores. Muscles were removed for identification of the unit with the PAS reaction and histochemical fiber typing by the use of modified standard techniques. 3. In the total population of MUs sampled, isometric tetanic force ranged from 5 to 441 mN in normal muscles and from 5 to 498 mN in reinnervated muscles, and the mean values were not significantly different. In the smaller sample of glycogen-depleted units from normal muscle, for a force range of 14-217 mN, N varied from 57 to 202, and A varied from 1,135 +/- 45 to 6,706 +/- 172 (SE) microns2. Within each unit the variation in fiber area is broad. After reinnervation, for a force range of 30-278 mN, N varied from 70 to 374, and A varied from 1,694 +/- 81 to 5,425 +/- 93 microns2. Mean fiber number was 153 +/- 18 in reinnervated muscle, which is significantly higher (P less than 0.01) than the normal value of 121 +/- 9. 4. The contribution of N and A to MU tetanic force was assessed by plotting each factor as a function of force on a log-log scale. N accounts for 39% and A for 49% of the variation in force in normal muscle. The contributions are changed after reinnervation where N, accounting for 65% of force, appears to compensate for the reduced range in A, which accounts for only 19% of the variation in force.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1992

Motor-unit categorization based on contractile and histochemical properties: a glycogen depletion analysis of normal and reinnervated rat tibialis anterior muscle.

1. Isolated and glycogen-depleted motor units (MUs) have been studied in normal and reinnervated tibialis anterior (TA) muscles of the rat to examine 1) the correspondence between physiological and histochemical classifications, 2) the extent to which unit properties cluster according to type, 3) the relation between unit force and fatigability, and 4) the extent to which reinnervated MUs recover their former properties. 2. MUs were isolated by ventral root dissection and stimulation in reinnervated and normal TA muscles, 3.5-8 mo after common peroneal (CP) nerve section and resuture and in age-matched control rats, respectively. The units were characterized physiologically for classification into four types: slow twitch (S), fast twitch, fatigue resistant (FR), fast twitch fatigue intermediate (FI), and fast twitch fatigue sensitive (FF). Four muscle fiber types were identified histochemically with the use of a modification of the techniques of Brooke and Kaiser, and Guth and Samaha to delineate fiber subtypes on the basis of the pH sensitivity of myofibrillar adenosine triphosphatase (ATPase). 3. Neither the time-to-peak twitch force development nor the profile of unfused tetanus ("sag test") was unambiguous in separating fast from slow MUs. However, all units with a time to peak greater than 22 ms were fatigue resistant, and this time was chosen to delineate fast from slow. The fast unit population was further subdivided on the basis of their fatigability. There is normally a small proportion of S units (6% S) that increased to 20% after reinnervation. Although the fast population was subdivided, there was a continuous distribution of fatigue indexes in normal and reinnervated muscles with the highest number of fast units falling into the FI category. The proportions of fast units were 28% FR, 45% FI, and 21% FF in normal muscles and 29% FR, 38% FI, and 13% FF in reinnervated muscles. 4. In normal muscles, delineation of fast and slow fibers and subdivision of fast fiber types on the basis of acid and alkali stability of myofibrillar ATPase provided a histochemical classification that showed 78% correspondence with physiological classification of the same identified units. In reinnervated muscles the correspondence between physiological and histochemical classifications was reduced to 72%. 5. The normal correlation between MU fatigability and isometric force in TA muscles was not seen in reinnervated muscles that contained more FR MUs. Mean fatigue index from normal units was significantly less at 0.55 +/- 0.03 (mean +/- SE) compared with 0.68 +/- 0.03 from reinnervated units.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1992

Proportional enlargement of motor units after partial denervation of cat triceps surae muscles.

1. To determine the capacity of motoneurons to increase their motor unit (MU) size by collateral sprouting and to assess this capacity in relation to the size of the motor nerve, we partially denervated soleus, lateral gastrocnemius (LG), and medial gastrocnemius (MG) muscles in adult and neonatal cats. Isometric force and extracellular nerve potentials were recorded from > or = 7% of the remaining MUs, 2.5-18 mo later. S1 or L7 roots were sectioned unilaterally and the number of remaining MUs was quantified by use of charge and force measurements. 2. The mean unit force increased inversely with MU number in partially denervated muscles, but the increase was less than predicted for extensive denervations (> or = 90%). The same enlargement of MU size occurred whether muscles were partially denervated in neonatal or adult animals. 3. The force distribution of MUs in partially denervated muscles was similar to normal but was shifted to larger force values in direct proportion to the extent of partial denervation (PD). All MUs increased in size by the same factor to preserve the normal force distribution. 4. Normal size relationships among force, contractile speed, and axon potential amplitude were observed for MUs in partially denervated muscles. Because changes in muscle fiber size could not account for the increase in unit force, these data show that increase in MU size, with respect to unit force and innervation ratio (muscle fibers per motoneuron), is proportional to the size of the motor nerve. 5. Enlargement of MU size in partially denervated muscles did not have a retrograde effect on nerve fiber caliber because axon potential amplitude and conduction velocity were not changed after PD. 6. Under conditions of extensive PD (> 85%), regenerated nerves from the cut spinal root reinnervated the gastrocnemius muscles. It is likely that nerves supplied muscle fibers that were not innervated by sprouts from nerves in the uncut root as well as displacing sprouts from terminals in extensively enlarged MUs. 7. We conclude that all motoneurons within a motor pool compensate for partial nerve injuries by collateral sprouting and that enlargement of MU size is a function of motor nerve size, consistent with Henneman's size principle.

Action Potentials↗

PubMed1992

Definition of a discontinuous immunodominant epitope at the NH2 terminus of the La/SS-B ribonucleoprotein autoantigen.

High-titer IgG autoantibodies to the La/SS-B ribonucleoprotein (RNP) are a hallmark of patients with primary Sjogren's syndrome. Anti-La/SS-B-positive human sera bind to multiple epitopes on recombinant La/SS-B, although the initial response is against an immunodominant epitope within the first 107 NH2-terminal amino acids (aa). Sequence analysis has identified a striking homology between aa 88-101 in this NH2-terminal region of La/SS-B and a feline retroviral gag polypeptide suggesting the anti-La/SS-B response may be initiated by cross-reactivity with an exogenous agent. In the present study, detailed mapping of this NH2-terminal epitope, using recombinant La/SS-B purified from the expression of overlapping DNA fragments spanning aa 1-107, has shown that this immunodominant epitope is a complex conformational or discontinuous epitope dependent upon both aa 12-28 and 82-99 for expression, even though these regions share no homology with each other. This requirement questions the significance of the homology between La/SS-B and a retroviral gag polypeptide in the generation of the B cell response to La/SS-B and is in accord with the general concept that B cells recognize conformational epitopes on antigens rather than small linear peptide sequences. The finding also reinforces the notion that native autoantigen could be the initiator of the autoimmune response.

Autoantibodies↗

PubMed1992

Molecular cloning and preliminary characterization of a novel cytoplasmic antigen recognized by myasthenia gravis sera.

A cDNA clone was isolated by screening of a lambda gt11 endothelial expression library with serum from a patient with myasthenia gravis (MG). Rabbit antisera raised against the recombinant protein and human MG sera reactive with the clone immunoblotted an M(r) integral of 250,000 polypeptide (gravin) present in endothelial cells and several adherent cells. Gravin was not detected in platelets, leukocytes, U937, or human erythroleukemic (HEL) cell lines, but was expressed in HEL cells after induction with phorbol myristate acetate. Northern blot analysis showed two transcripts of approximately 6.7 and 8.4 kb in endothelial cells but not U937 or HEL cells. Indirect immunofluorescence of permeabilized cells revealed a trabecular network of gravin staining with a distinct linear component. Antibodies to gravin, were present in sera from 22:72 (31%) of MG patients. In contrast 0:50 normal sera and 1:72 sera from patients with other autoimmune diseases contained antigravin antibodies. Gravin is not likely to be a nonerythroid spectrin, talin, myosin, or actin-binding protein based on the lack of reactivity of antigravin with these polypeptides in immunoblots. The nucleotide sequence of the immunoreactive clone indicated that it encodes a highly acidic polypeptide fragment that contains the carboxyl terminus of the protein. Neither amino acid nor nucleotide sequences were present in Genbank, EMBL, or Swissprot databases as of March, 1992. These data indicate that gravin is an inducible, cell type-specific cytoplasmic protein and that auto-antibodies to gravin may be highly specific for MG.

Amino Acid Sequence↗

PubMed1992

Incidence and cost of hospital admissions secondary to drug interactions involving theophylline.

OBJECTIVE: To determine the incidence and cost of hospital admissions for theophylline toxicity, which occurred as a result of the concurrent use of one of the following medications: cimetidine, erythromycin, or ciprofloxacin. DESIGN: Retrospective chart review (18 months, between June 1989 and November 1990). SETTING: A Department of Veterans Affairs Medical Center. PARTICIPANTS: All patients who were receiving theophylline chronically (913 patients) and also had a prescription for cimetidine (124 patients with 140 treatment courses), erythromycin (66 patients with 93 treatment courses), or ciprofloxacin (39 patients with 59 treatment courses) dispensed. INTERVENTIONS: Each patient's medical record was reviewed to identify hospital admissions within 30 days following the dispensing of the interacting drug. MAIN OUTCOME MEASURES: Admissions were considered to be related to theophylline toxicity if appropriate signs and symptoms were present and the theophylline concentration was above 20 micrograms/mL or had increased significantly from the concentration obtained prior to introduction of the interacting drug. RESULTS: One patient who received cimetidine and one who received ciprofloxacin were admitted for theophylline toxicity (2 of 292 potential interactions, 0.81 percent). Admissions were for 16 and 13 days, respectively, and total costs for the two admissions were $12,864.22 or $44.00, respectively, per potential interaction. The entire admission was not for theophylline toxicity; it appeared that iatrogenic factors contributed to the duration. CONCLUSIONS: The incidence of hospital admissions secondary to theophylline drug interactions with cimetidine, ciprofloxacin, or erythromycin is low, but the admissions represent considerable expense, even when distributed among all patients at risk for the interactions.

Aged↗