State of the Journal: 1999

Maternal and Child Health Journal -     

Treatment of autoimmune thrombocytopenic purpura with rhesus antibodies (anti-Rh0(D))

Summary There is evidence that blockade of the reticuloendothelial system (RES) by sequestration of autologous red blood cells (RBC) leads to an elevation of platelet counts in immune thrombocytopenia. To substantiate this hypothesis, 10 Rh₀(D)-positive adult patients (9 female, 1 male) with chronic autoimmune thrombocytopenic purpura (ITP) (1 to 21 years duration) were treated with low doses of intravenous IgG-anti-Rh₀(D) (200 to 1,000 g per dose; 300 to 3,600 g per course; administration within 1 to 5 days). All patients improved clinically as indicated by cessation of bleeding. In eight out of ten patients there was a rise in platelet count. Platelet increments were excellent (>100×10⁹/l) in one, good (50–100×10⁹/l) in three, fair (20–50×10⁹/l) in two and low (10–20×10⁹/l) in two patients. Splenectomized patients (N=4) had a poorer response than non-splenectomized patients (N=6) with mean increments of 16×10⁹/l (range 5–43×10⁹/l) versus 60×10⁹/l (range 10–110×10⁹/l). The increase in platelet counts persisted for seven to over 150 days. Transient and slight signs of haemolysis developed in seven out of ten patients (haemoglobin remained stable; increase of lactate dehydrogenase (>250 IU/l) in four, decrease of haptoglobin (<60 mg/dl) in five patients). The direct antiglobulin test became positive in all cases due to IgG1 without complement fixation. We conclude that the interaction of antibody-coated RBC with macrophages (and, probably, other means of RBC alteration) is a feasible therapeutic approach in selected cases of ITP and related conditions.     

Anomalous migration of central nervous tissue to transplanted autonomic ganglia

Summary When fragments of adult superior cervical ganglion (SCG) were transplanted to the undamaged surface of the cerebellum of 6–14-day-old rats, a series of remarkable changes ensued that were apparent during the postoperative periods of three weeks to two years: external granule cells were either arrested at the brain surface in the form of ectopic laminae, or migrated anomalously out of the brain to invade the SCG graft. Not only were granule cells so affected, but other interneurons, such as basket and stellate cells as well. The associated neuropil itself became displaced. Thus, mossy fibre boutons formed synaptic glomeruli with dendrites of arrested granule cells at the cerebellar surface. Reactive astrocytes, forming either thin parallel sheets or thick processes, and packed with filaments, mingled closely with the arrested neurons; typical Bergmann glia were not observed. Extensive bridges from the cerebellum to the SCG contained not only synaptically mature interneurons but also rows of parallel fibres, mossy fibres forming mini-glomeruli with solitary granule cells, and reactive astroglia. Portions of the Purkinje arbor extended into the tissue bridges and received synapses on elongate spines from co-migrating parallel fibres. Astroglial cells received synapse-like contacts on their somata and processes. Similar CG grafts induced tissue bridges, predominantly astroglial, to arise from the area postrema or dorsal medulla and enter the graft.Control transplantations tested the specificity of these cellular reactions. Other tissue grafts included muscle, salivary gland and normal or pre-degenerated sciatic nerve. Only direct allografts of sciatic nerve appeared to cause displacement of scattered granule cells. Non-biological grafts such as dacron, silicone and polystyrene, while causing mechanical deformation and inflammation, did not alter normal granule cell migration.The anomalously migrating neurons and neuropil, which seem to have by-passed certain patterns of cellular development, may respond positively to some diffusible tropic substances. The further determination of certain influences that may elicit these unique tissue-tissue interactions, suggested in the framework of this study to be components of neural regeneration or degeneration, could elucidate some aspects of neuronal interactions.     

DUAL CHART DRIVE FOR GRASS POLYGRAPH

A modification of a polygraph in which a second chart drive is incorporated to allow simultaneous recording at two different chart speeds is described. The auxiliary chart drive is of the same type as the original, having ten speeds and operating independently of the original. Specific uses of the two-chart system are discussed.     

Polymorphisms in the 5' region of the CD14 gene are associated with eczema in young children

BACKGROUND: Variants in the CD14 gene (CD14) are hypothesized to be associated with atopic disorders. However, most studies have only investigated one polymorphism in this gene. OBJECTIVE: We sought to study the association of 5 single nucleotide polymorphisms (SNPs) in the 5' flanking region of CD14 with eczema and serum IgE levels in young children. METHODS: We genotyped 5 SNPs in an approximately 6.5-kb region in the 5' region of CD14 in 344 2-year-old white children from 2 birth cohorts in the northeastern United States. We examined the relation of both single SNPs and haplotypes in CD14 with the atopic outcomes. RESULTS: Two SNPs were significantly associated with eczema. In dominant models adjusted for potential confounders, SNP rs2569193 was associated with significantly decreased risk for eczema (odds ratio [OR] for CT/TT vs CC, 0.5; 95% CI, 0.3-0.8), whereas SNP rs2569190 (also reported as the C-159T) was associated with significantly increased risk for eczema (OR for CT/TT vs CC, 2.3; 95% CI, 1.4-3.8). The CT/TT genotypes of SNP rs2569190 also had higher geometric means of serum IgE than the CC genotype (24.6 vs 15 IU/mL, P = .025). Haplotype analyses provided results similar to those of the single SNP analyses. CONCLUSIONS: Our results contradict previous reports that have found a protective effect of the T allele of SNP rs2569190 (C-159T) against atopic disorders. Nevertheless, these results confirm the importance of polymorphisms in CD14 in the development of atopy, and future studies of this gene region will need to account for linkage disequilibrium and environmental exposures unique to the study population.     

THE EFFECTS OF AWARENESS ON CORTICAL EVOKED POTENTIALS TO CONDITIONED AFFECTIVE STIMULI

A previous paper of ours (Begleiter, Gross, & Kissin, 1967) demonstrated that it was possible to condition affective meaning to meaningless figures (CS), and significantly alter visual evoked potential (VEP) amplitudes and latencies to them, without the S's awareness of the CS–UCS relationship (Experiment I, totally unaware). In the present study some Ss were deliberately informed that a CS–UCS connection existed; however, the exact nature of their relationship was not divulged (Experiment II, slightly aware). Other Ss were explicitly informed of the correct CS–UCS contingency, and entire conditioning paradigm (Experiment III, fully aware). One physiological (VEP) and two behavioral (interflash interval and semantic differential) indices of conditioning were obtained during an extinction procedure, and demonstrated significant differences between CRs in Experiment II, but none in Experiment III. VEP amplitudes to positive and negative CSs were enhanced in Experiment II, and suppressed in Experiment I, in comparison to the neutral CS. This effect was most marked in responses to the negative CS. It is suggested that level of awareness of the CS–UCS contingency might be reflected in our physiological index of conditioning - VEP amplitude.     

Immunosuppressive activity of FK-506 in rats: flow cytometric analysis of lymphocyte populations in blood, spleen and thymus during treatment and following drug withdrawal.

Rats were immunized systemically with sheep red blood cells (SRBC) and given either FK-506 (1 mg/kg) or drug vehicle by i.m. injection for 7 days. In animals receiving FK-506, there was suppression (87%) of the splenic plaque-forming cell response on day 4 and marked reductions in the serum antibody titre throughout the 3-week period following immunization. Sequential flow cytometric analyses of blood lymphocytes revealed statistically significant attenuation, by FK-506, of the increase in relative numbers of OX-12+ (B) cells between days 4 and 7. Following drug withdrawal, OX-12 values remained elevated, whereas in control animals a decline was observed. These changes were reflected in concomitant increases in the relative numbers of OX-19+ (CD3+), W3/25+ (CD4+) and OX-8+ (CD8+) T cells; however, due to an overall reduction in lymphocytes by day 7, absolute values were not significantly affected compared with controls. The pattern of changes in OX-6+ (MHC class II+) cells in blood was similar to that observed for B cells. FK-506 also suppressed increases in the small proportion of blood-borne OX-40+ (activated CD4+) cells and OX-39+ (interleukin-2 receptor+) cells in the 7 day period following immunization; thereafter values for activation marker expression between treatment and control groups were similar. In the spleen, there were fewer significant differences between FK-506 and control groups in the incidences of cells expressing the above markers. OX-8+ cells, however, were significantly higher in drug-treated animals on day 7, and there were also reductions in the small proportions of OX-39+ and OX-40+ cells when compared with controls. In the thymus, reversible medullary atrophy induced by FK-506 was accompanied on day 7 by increases in the incidence of CD4+ and CD8+ cells and by a concomitant reduction in OX-44+ mature, medullary thymocytes. Two weeks after drug withdrawal, the phenotypic marker expression profile had been restored to normal in blood, spleen and thymus. These data provide new information on the apparent capacity of FK-506 to interfere with T cell maturation and its influence on lymphocyte activation in vivo.     

In vitro and in vivo translation of the ribonucleic acids of a cowpea strain of tobacco mosaic virus1

Experiments reported here support the notion that intact TMV RNA has genes which are not translated directly and that specific fragments of the RNA are the functional messengers for those genes.A discrete class of short nucleoprotein rods, in addition to the expected 300-nm rods, was observed by Morris in a cowpea strain of TMV isolated from infected cowpea tissue. We have found a third class of rods of intermediate length. RNAs from the three kinds of rods were separately translated in a wheat germ cell-free protein synthesis system. RNA from the short rods directed the synthesis of a polypeptide with some properties of capsid protein: the same electrophoretic mobility in detergent-impregnated polyacrylamide gels and the ability to participate in a reconstitution reaction with virus RNA. RNA from long rods directed the synthesis of a spectrum of polypeptides. Among these one of ～130,000 molecular weight predominated. It had the same electrophoretic mobility as a polypeptide which was recovered from virus-infected tissue, but not from mock-inoculated tissue. RNA from rods of an intermediate length class directed the synthesis of one or two polypeptides with apparent molecular weights of about 30,000. Capsid polypeptide could not be demonstrated among the products formed under the direction of the long or immediate rods RNAs. However, RNA-RNA hybridization experiments indicate that short and intermediate rod RNAs share nucleotide sequences with each other and with the long rod RNA. The possible relationships of these results to the replication of the common strain of TMV and brome mosaic virus are discussed.     

Monoclonal antibodies recognizing CD5, CD10 and CD23 in formalin-fixed, paraffin-embedded tissue: production and assessment of their value in the diagnosis of small B-cell lymphoma

AIMS: Assessment of the expression of antigens CD5, CD10 and CD23 can be of value in the differential diagnosis of small B-cell lymphoma. Correct subclassification is important since optimal treatment regimes differ between the subtypes. The aim of this study was to generate monoclonal antibodies recognizing these antigens in paraffin-embedded tissue and to assess their efficacy using a panel of cases of small B-cell lymphoma of various subtypes. METHODS AND RESULTS: For each antibody synthetic recombinant protein and conventional murine hybridoma technology was employed. Monoclonal antibodies effective in formalin-fixed, paraffin-embedded tissue were successfully generated, designated NCL-CD5-4C7, NCL-CD10-270 and NCL-CD23-1B12, respectively. A series of 58 cases of small B-cell lymphoma including examples of each subtype (lymphocytic, follicle centre cell, mantle cell, marginal zone and lymphoplasmacytoid) was assembled and immunostaining for the respective antigens carried out using the monoclonal antibodies produced. Our results indicate that the antibodies are specific for their respective antigens and give the predicted phenotypic profile in the small B-cell lymphoma subtypes. CONCLUSIONS: These novel monoclonal antibodies may be of value in routine diagnostic practice.