A longitudinal study of pulmonary involvement in primary Sjogren's syndrome: relationship between alveolitis and subsequent lung changes on high-resolution computed tomography

Eighteen non-smoking women suffering from primary Sjogren's syndrome (pSS) with previously documented alveolitis were re-examined, clinically and by pulmonary function tests (PFT), bronchoalveolar lavage (BAL), chest X-ray and high-resolution computed tomography (HRCT) after a 2 yr follow-up period. Longitudinal evaluation revealed unchanged PFT. The final BAL study showed a normal differential count in six of 14 patients with initial lymphocyte alveolitis, and a persistent alveolar lymphocytosis in the remaining eight patients, associated with an increased percentage of neutrophils in one of them. In four patients with initial mixed alveolitis, the BAL cell profile was unchanged 2 yr later. Five of 18 patients (28%) had abnormal HRCT, represented by isolated septal/subpleural lines in three patients, ground-glass opacities with irregular pleural margins in one patient, and ground-glass opacities associated with septal/subpleural lines in another. All these patients had abnormal BAL results with an increased proportion of both neutrophils and lymphocytes. The presence of alveolar neutrophils was associated with a significantly (P=0.005) greater mean rate of reduction of carbon monoxide diffusing capacity (DLCO) -- more than four times the normal rate of loss of DLCO. Chest X- ray, repeated at the end of the 2 yr follow-up period, showed parenchymal abnormalities in only one patient who had evidence of fibrosis on HRCT. This study provides evidence that lung involvement is not an uncommon extraglandular manifestation of pSS and that a BAL neutrophilia may play an important role in the pathogenesis of pulmonary disease in this autoimmune disorder.      

Chickenpox monoarthritis: demonstration of varicella-zoster virus in joint fluid by polymerase chain reaction

A case of chickenpox monoarthritis is described. The presence of varicella zoster virus (VZV) within the joint was demonstrated by the detection of viral DNA in synovial fluid at a time when peripheral blood cells were negative. This strongly suggests a direct role of VZV in causing monoarthritis complicating chickenpox. The use of the polymerase chain reaction allows more rapid (2 days) confirmation of the diagnosis. Early enough diagnosis would raise the question of using acyclovir to shorten the duration of arthritis.      

Characterization of a factor IX variant with a glycine207 to glutamic acid mutation

Factor IXTaipei9 is a factor IX variant from a hemophilia B patient with reduced levels of circulating protein molecules (cross-reacting material reduced, CRM). This variant contained a glycine (Gly) to glutamic acid (Glu) substitution at the 207th codon of mature factor IX. The functional consequences of the Gly-->Glu mutation in factor IXTaipei9 (IXG207E) were characterized in this study. Plasma-derived IXG207E exhibited a mobility similar to that of normal factor IX on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its specific activity was estimated to be 3.5% that of the purified normal factor IX in a one-stage partial thromboplastin time assay (aPTT). Cleavage of factor IXG207E by factor XIa or factor VIIa-tissue factor complex appeared to be normal. When the calcium-dependent conformational change was examined by monitoring quenching of intrinsic fluorescence, both normal factor IX and IXG207E exhibited equivalent intrinsic fluorescence quenching. Activated factor IXG207E (IXaG207E) also binds antithrombin III equally as well as normal factor IXa. However, aberrant binding of the active site probe p-aminobenzamidine was observed for factor XIa-activated factor IXG207E, indicating that the active site pocket of the heavy chain of factor IXaG207E was abnormal. Moreover, the rate of activation of factor X by factor IXaG207E, as measured in a purified system using chromogenic substrates, was estimated to be 1/40 of that of normal factor IXa. A computer-modeled heavy-chain structure of factor IXa predicts a hydrophobic environment surrounding Gly-207 and this Gly forms a hydrogen bound to the active site serine-365. The molecular mechanism of the Gly-->Glu mutation in factor IXTaipei9 might result in the alteration of the microenvironment of the active site pocket which renders the active site serine-365 inaccessible to its substrate.     

Marrow transplantation from hepatitis C virus seropositive donors: transmission rate and clinical course

Bone marrow transplant recipients are at risk for acquiring hepatitis C infection from the donated marrow. Twelve patients who were hepatitis C virus (HCV) RNA-negative pretransplant received marrow from anti-HCV seropositive donors. HCV RNA was present in the sera of seven of these donors. After transplant, serial serum specimens were obtained from all marrow recipients for determination of HCV RNA and aminotransferase levels. All seven recipients of marrow from HCV RNA-positive donors were HCV RNA-positive after marrow infusion; none cleared virus from the serum. All five recipients of marrow from anti-HCV seropositive, HCV RNA-negative donors remained free of HCV RNA in serum up to day 100. Abnormal serum aminotransferases were common in both HCV RNA- negative and HCV RNA-positive marrow recipients. One HCV-infected recipient developed marked elevation in aminotransferases after immunosuppressive drugs were stopped. We conclude that the presence of HCV RNA in the serum of marrow donors is an accurate predictor of HCV infection in marrow recipients. The acute infection was subclinical in all patients. The long-term risk of chronic hepatitis C virus infection in these patients remains to be determined.     

Retrovirus-mediated reconstitution of respiratory burst activity in X- linked chronic granulomatous disease cells

X-linked chronic granulomatous disease (X-CGD) results from mutations in the gene encoding gp91phox, the larger subunit of the respiratory burst oxidase cytochrome b. In this study, a recombinant retrovirus vector was constructed and evaluated for its expression of human gp91phox in a human X-CGD myeloid cell line in which the endogenous gp91phox gene had been disrupted by gene targeting. The retrovirus construct, Zip/PGKgp91, was first introduced into the GP+envAm12 amphotropic packaging line and yielded virus producer clones with estimated titers of up to 1 x 10(5) cfu/mL. Coculture infection of X- CGD myeloid cells with Zip/PGKgp91 resulted in restoration of respiratory burst activity to 15% of the cells. Isolated clonal infectants expressed relatively low levels of recombinant gp91phox (< or = 12% of wild-type), but exhibited considerable superoxide- generating activity (up to nearly 60% of wild-type). These results show the feasibility of phenotypic correction of CGD using gene replacement therapy and suggest that even modest levels of gp91phox expression may lead to considerable functional correction of X-CGD neutrophils.     

Comparison of Digital Versus Metacarpal Blocks for Repair of Finger Injuries

Study objective: This study compared efficacy, degree of discomfort, and time to anesthesia of digital blocks and metacarpal blocks for digital anesthesia. Design: Randomized, prospective, nonblinded, clinical study conducted from April 1992 to January 1993. Patients served as their own controls. Setting: Inner-city and community hospital emergency departments. Type of participants: Convenience sample of 30 adult patients, with third or fourth finger injuries including and distal to the proximal interphalangeal joint that required digital anesthesia. Interventions: Digital blocks and a metacarpal blocks were performed (one per side) on all 30 patients (total of 60 blocks). The order of the blocks was randomized. Measurements: A digital block and a metacarpal block were performed on each patient. Patients immediately rated the pain associated with each technique on a nonsegmented visual analog scale. Efficacy was assessed by requirement for additional anesthesia and anesthesia to pinprick. Time to anesthesia was assessed after each block in 23 patients. Results: Mean visual analog scale pain scores were 2.53 for digital block and 3.38 for metacarpal block (P = .1751, Student's t-test). Metacarpal block failed anesthesia to pinprick in 23% of patients compared to 3% for digital block (P = .0227, χ² ). Time to anesthesia was significantly shorter for digital block compared to metacarpal block, with a mean of 2.82 minutes versus 6.35 minutes (P < .0001, Student's t-test). Conclusion: Digital block and metacarpal block, as described in this study, are equally painful procedures. Digital block, however, is more efficacious and requires significantly less time to anesthesia for the injured finger.[Knoop, K, Trott A, Syverud S: Comparison of digital versus metacarpal blocks for repair of finger injuries. Ann Emerg Med June 1994;23:1296-1300.]     

Tachykinin Actions on Deep Dorsal Horn Neurons In Wm: An Electrophysiological and Morphological Study in the Immature Rat

To assess whether functional neurokinin receptors exist in the deep dorsal horn of the rat, the actions of the selective neurokinin-1 receptor (NK1R) agonist [Sar⁹,Met(O₂)¹¹]substance P [Sar⁹,Met(O₂)¹¹]]SP), the neurokinin-2 receptor (NK2R) agonists [β-Ala⁸]NKA_4-10 and GR64349 and the neurokinin-3 receptor (NK3R) agonist senktide were examined intracellularly in vitro. [Sar⁹,Met(O₂)¹¹]]SP (1–4 μM) and senktide (1-2 μM) elicited slow depolarizations (40 mV) associated with increased synaptic activity and cell firing. [β-Ala⁸]NKA_4-10 (10-20 μM) and GR64349 (0.25-10 μM) caused small depolarizations (<2.0 mV) and no firing. Neurons were categorized as either ‘tonic’ or ‘phasic’ depending on their firing response to direct current step depolarizations. Tonic neurons, which, unlike phasic neurons, display no spike firing accommodation, generated a significantly larger depolarization to the NK1R and NK3R agonists. The putative contribution of these receptors to primary afferent-mediated synaptic transmission was assessed by testing the NKIR antagonist GR82334 (1 μM), the NK2R antagonist MEN10,376 (1 μM) and the NK3R antagonist [Trp⁷,β-Ala⁸]NKA4_-10 (1 μM) against the dorsal root-evoked excitatory postsynaptic potential (DR-EPSP). GR82334 and [Trp⁷,β-Ala⁸]NKA_4-10 significantly reduced (P ≤ 0.05) the duration but not the amplitude of the DR-EPSP. MEN10,376 (1 μM) had no effect on DR-EPSP amplitude or duration. Morphological detail was obtained for seven biocytin-filled deep dorsal horn neurons tested with [Sar⁹,Met(O₂)¹¹]SP. Five neurons responded to the NKIR agonist, and two of these had dorsally directed dendrites into the substantia gelatinosa. The other three [Sar⁹,Met(O₂)¹¹]SP-sensitive neurons had dendrites within deeper laminae. These data support the existence of functional NK1Rs and NK3Rs in the deep dorsal horn which may be involved in mediating sensory afferent inputs from nociceptors.     

Effect of L-dopa loading on 5-HTP decarboxylation in rat brain areas

The time course of 5-hydroxytryptophan (5-HTP), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) concentrations in four rat brain areas (hypothalamus, hippocampus, striatum and olfactory bulbs) were investigated after treatment with L-dopa (125 mg/kg, ip) + benserazide (50 mg/kg, ip). 5-HTP levels increased as early as 0.5 h, showed maximum accumulation at 1.5 h and returned to control levels within 4 h, while 5-HT was markedly decreased in all four structures, with a maximum effect at 1.5 h (approximately -70%) in the four areas. The decrease in 5-HT was not accompanied by changes in 5-HIAA levels. In agreement with previous studies, these data demonstrate that L-dopa loading interferes with serotonin metabolism in the rat brain. However, in addition to the releasing action of newly-synthesized dopamine, the accumulation of 5-HTP and the parallel decrease in 5-HT indicate a reduction in 5-HT synthesis. This inhibition could be explained by a competitive effect of L-dopa for aromatic aminoacid decarboxylase activity.     

Polymerase chain reaction-based strain characterization of noncapsulate Haemophilus influenzae.

A polymerase chain reaction-based typing method for noncapsulate Haemophilus influenzae was developed. Randomly amplified polymorphic DNA fingerprints were generated from boiled supernatants prepared directly from bacterial colonies without the need for DNA extraction. The technique was applied to isolates obtained during putative outbreaks of chest infection and validated by comparison with sodium dodecyl sulfatepolyacrylamide gel electrophoresis analysis of outer membrane protein-enriched preparations and rRNA gene restriction analysis. There was complete concordance between the three techniques. The results show that randomly amplified polymorphic DNA analysis provides a highly discriminatory method of characterizing strains of noncapsulate H. influenzae which is eminently suitable as an epidemiological tool for the rapid investigation of outbreaks of infection.     

The preferential effects of chloroquine on the IgM and IgG subclass responses to TI and TD antigens in BALB/cAn mice

This study has determined the effects of chloroquine on the IgM and the IgG subclass responses of BALB/cAn mice to both thymus-independent (TI) and thymus-dependent (TD) antigens, and has found that in antigen-unprimed BALB/cAn mice chloroquine adversely affects antibody responses to both TI and TD antigens. However, in antigen-primed mice the immune responses to TD antigens were unaffected by chloroquine. The IgGl subclass response, but not necessarily other IgG subclass or IgM responses to TI and TD antigens, was adversely affected by chloroquine in unprimed mice only. Thus, in unprimed, but not in antigen-primed, BALB/cAn mice, chloroquine preferentially decreased the IgGl subclass responses to both TI and TD antigens.