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Reproduction has been shown to be costly for survival in a wide diversity of taxa. The resulting trade-off, termed the reproduction-survival trade-off, is thought to be one of the most fundamental forces of life-history evolution. In insects the pleiotropic effect of juvenile hormone (JH), antagonistically regulating reproduction and pathogen resistance, is suggested to underlie this phenomenon. In contrast to the majority of insects, reproductive individuals in many eusocial insects defy this trade-off and live both long and prosper. By remodelling the gonadotropic effects of JH in reproductive regulation, the queens of the long-lived black garden ant Lasius niger (living up to 27 years), have circumvented the reproduction-survival trade off enabling them to maximize both reproduction and pathogen resistance simultaneously. In this study we measure fertility, vitellogenin gene expression and protein levels after experimental manipulation of hormone levels. We use these measurements to investigate the mechanistic basis of endocrinological role remodelling in reproduction and determine how JH suppresses reproduction in this species, rather then stimulating it, like in the majority of insects. We find that JH likely inhibits three key aspects of reproduction both during vitellogenesis and oogenesis, including two previously unknown mechanisms. In addition, we document that juvenile hormone, as in the majority of insects, has retained some stimulatory function in regulating vitellogenin expression. We discuss the evolutionary consequences of this complex regulatory architecture of reproduction in L. niger, which might enable the evolution of similar reproductive phenotypes by alternate regulatory pathways, and the surprising flexibility regulatory role of juvenile hormone in this process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Objectives: This in vitro study investigated the effect of selected luting media on the fracture resistance of a flame-sprayed all-ceramic crown., Methods: Three groups of 10 human upper premolar teeth were prepared for crowning using a standardised technique. Flame sprayed crowns were fabricated and cemented onto the preparations using zinc phosphate (ZPC), glass polyalkenoate (GPC) or composite luting cement (CLC). During crown seating, a pressure perfusion system simulated pulpal fluid outflow equivalent to 300mm of H2O. Compressive fracture resistance was determined for each group using a Universal Testing Machine with a crosshead speed of 1mm min(-1). A group of unrestored teeth acted as a control., Results: The fracture resistance of the groups ranked as follows: ZPC>CLC>>GPC=unrestored teeth. The difference between the fracture resistance of ZPC and CLC groups and the control group was statistically significant. The mode of fracture between the luted crowns and natural crowns was markedly different., Conclusions: When tested in compression, a new, flame-sprayed all-ceramic crown, when luted in place using ZPC, GPC or CLC, could produce strengths comparable to or greater than natural unrestored teeth. The luting agent used significantly affected the recorded fracture loads.

Objectives: The aims of this in vitro study were to (a) measure fluid flow through teeth restored with one of three dentine bonding systems and a resin composite restoration; (b) measure the distribution of a silver tracer through the same teeth, and make a comparison with fluid flow; and (c) investigate the effect of thermocycling on both measurement types., Methods: Coronal segments of 30 premolars, randomly allocated to three equal groups, were assessed by fluid filtration. Each group was restored with a resin composite restoration in conjunction with Fuji Bond LC (FBLC), Scotchbond Multi-Purpose Plus (SMP+) or Prime&Bond 2.1 (P&B2.1). Fluid filtration rates were measured in the intact crown and then after cavity preparation, conditioning, dentine bonding, restoration and at 2 and 24 h, 1 week and 1 month following restoration. Six specimens from each group were thermocycled at 1 week. After final filtration measurements the specimens were perfused with silver nitrate and cleared before scoring tracer penetration., Results: No significant differences (P > 0.05) in fluid filtration rates were found amongst the different bonding systems or at any restoration stage. Thermocycling was not associated with any significant (P > 0.05) increase in fluid filtration. Final fluid filtration and tracer distribution showed a weak and not statistically significant correlation (P > 0.05). The penetration of silver stain indicated a failure of the restorations to seal the cavity and demonstrated a possible pathway by which in vivo post-operative sensitivity could occur., Conclusions: Although not statistically significant, conditioning of the cavity increased the dentinal permeability but this effect was variable. Thermocycling had no statistically significant effect on microleakage.

Type VII collagen, the major component of anchoring fibrils, consists of a central collagenous triple-helical domain flanked by two noncollagenous, globular domains, NC1 and NC2. Approximately 50% of the molecular mass of the molecule is consumed by the NC1 domain. We previously demonstrated that NC1 binds to various extracellular matrix components including a complex of laminin 5 and laminin 6 (Chen et al, 1997a). In this study, we examined the interaction of NC1 with laminin 5 (a component of anchoring filaments). Both authentic and purified recombinant NC1 bound to human and rat laminin 5 as measured by enzyme-linked immunosorbant assay and by binding of 125I-radiolabeled NC1 to laminin 5-coated wells, but not to laminin 1 or albumin. NC1 bound predominantly to the beta3 chain of laminin 5, but also to the gamma2 chain when examined by a protein overlay assay. The binding of 125I-NC1 to laminin 5 was inhibited by a 50-fold excess of unlabeled NC1 or de-glycosylated NC1, as well as a polyclonal antibody to laminin 5 or a monoclonal antibody to the beta3 chain. In contrast, the NC1-laminin 5 interaction was not affected by a monoclonal antibody to the alpha3 chain. Using NC1 deletion mutant recombinant proteins, a 285 AA (residues 760-1045) subdomain of NC1 was identified as the binding site for laminin 5. IgG from an epidermolysis bullosa acquisita serum containing autoantibodies to epitopes within NC1 that colocalized with the laminin 5 binding site inhibited the binding of NC1 to laminin 5. Thus, perturbation of the NC1-laminin 5 interaction may contribute to the pathogenesis of epidermolysis bullosa acquisita.

Cell adhesion to extracellular matrix contributes to the organization of tissues and modulates cell behavior. In conventional cell adhesion assays, plastic wells are coated with matrix proteins and assayed for their adhesion-promoting activity. We show here that factors such as sample composition, coating buffers, and manufacturers' plastic treatment markedly affect cell adhesion to the extracellular matrix protein laminin-5 (Ln-5). These factors were shown to affect adsorption efficiency as determined by measuring total adsorbed protein with a polyclonal anti-Ln-5 antiserum. They also influence the availability of the epitope for an adhesion-blocking anti-Ln-5 monoclonal antibody, suggesting that coating conditions affect the orientation of Ln-5. Generally, cell adhesion correlates more strongly with the availability of the epitope for the adhesion-blocking antibody than with total adsorbed Ln-5. Our data further indicate that cell adhesion to other matrix proteins may be influenced by similar factors. Adding Ln-5 samples to plastic wells that had been precoated with non-adhesion-blocking anti-Ln-5 antibodies made cell adhesion independent of factors such as sample composition, coating buffers, and source of plastic. Thus, the control of adsorption efficiency and orientation of extracellular matrix proteins is essential for creation of reliable and reproducible conditions in cell adhesion assays.

The hemidesmosome is a multimolecular complex that integrates the extracellular matrix with the keratin cytoskeleton and that stabilizes epithelial attachment to connective tissue. A 180 kDa protein (BP180, type XVII collagen), first identified by its reactivity with autoantibodies in the serum of patients with a blistering skin disease called bullous pemphigoid (BP), is a transmembrane component of the hemidesmosome with a collagen-like extracellular domain. Here, using recombinantly expressed molecules and the yeast two-hybrid assay, we have identified alpha6 integrin as a BP180-binding partner. The association between specific domains of the BP180 and alpha6 integrin molecules is inhibited by a 14 mer peptide, whose sequence is identical to amino acid residues 506-519 in the noncollagenous region of the ectodomain of the BP180 molecule, as well as by antibodies raised against this peptide. The 14 mer peptide sequence is part of an epitope recognized by autoantibodies that are pathogenic in BP. In vivo, when 804G cells are plated into medium containing the same peptide, they fail to assemble hemidesmosomes. Furthermore, although BP180 and certain cytoplasmic components of the hemidesmosome colocalize in the peptide-treated cells, they are aberrantly distributed and fail to show extensive association with (alpha6beta4 integrin. Taken together, our results indicate that BP180 is a novel transmembrane ligand of the alpha6beta4 integrin heterodimer. In addition, our data provide support for the possibility that BP180 and alpha6 integrin interaction is not only mediated by the BP epitope but is necessary for hemidesmosome formation.

The advantage of using sphenoidal (SE) over antero-temporal electrodes (ATE) remains controversial among epileptologists. Yet, in a recently published study of 17 patients with seizures of antero-temporal origin (Kanner et al., 1995), we demonstrated that SE placed under fluoroscopic guidance (FPSE), in order to insure that their recording tips are positioned immediately below the foramen ovale (FO), yielded a significant advantage over SE placed with the standard blind method of insertion (BPSE), in both interictal and ictal recordings. This study was done to test the following hypothesis: FPSE advantage over BPSE and ATE resides in the recording of epileptiform activity with a restricted electric field. We compared spike voltages at FPSE, BPSE and ATE in sets of 5 randomly selected spikes per interictal focus, recorded in the course of separate monitoring studies with BPSE and FPSE. We represented the voltage differences as ratios, VATE/FPSE and VATE/BPSE and calculated a mean ratio for each spike set. The spikes' voltage was almost identical at BPSE and at ATE (mean VATE/BPSE = 0.94), while it was significantly higher at FPSE than at ATE (mean VATE/FPSE = 0.66; P < 0.001, t test). A significantly narrower electric field contour was found among interictal foci in which FPSE yielded additional data during interictal (P < 0.001) and ictal (P = 0.016) recordings. Conversely, VATE/FPSE did not differ from VATE/BPSE among interictal foci where FPSE failed to yield any advantage over BPSE in either interictal (P = 0.240), or ictal (P = 0.311) recordings. These findings prove that SE yield additional localizing data when recording epileptiform activity with a restricted field, provided that its recording tip is positioned below the FO. When distant from FO, SE can be expected to yield comparable data to that obtained with ATE.

Hemidesmosomes are multiprotein structures that attach basal cells of stratified epithelia to basement membranes. Although normal human breast epithelia are not stratified, we observed expression of electron-dense hemidesmosomes and hemidesmosome protein components by breast epithelial and myoepithelial cells at the basal lamina in vivo. Primary cultured normal human breast epithelial cells also contained hemidesmosomes and component proteins, and could be used as a model for hemidesmosome assembly and regulation. In these cultured cells, hemidesmosome proteins were expressed and localized basally in an unvaried temporal pattern, and electron-dense hemidesmosomes were not seen until the final protein was localized to the cell base. In addition, rate of localization was influenced by confluence, doubling time, and extracellular matrix. Invasive breast cancer cells did not express hemidesmosomes or most of the component proteins in vivo. In carcinoma in situ, cells away from the basement membrane lacked hemidesmosomes and hemidesmosome proteins, and cells at the basement membrane exhibited abnormalities of hemidesmosome protein expression. Primary human malignant breast cells in culture exhibited a mix of hemidesmosome phenotypes. These data suggest that hemidesmosomes may be important subcellular structures in determining the cytoarchitecture of the breast epithelium. Further, their downregulation may influence cytoarchitecture remodeling closely linked with cell cycle, motility, and extracellular matrix interactions; and their loss in carcinoma may be associated with loss of normal cytoarchitecture.

HaCaT cells, an immortalized keratinocyte line, incubated in plastic wells in the presence of conditioned medium from 804G cells adhered and spread rapidly in less than 30 min. In contrast, cells plated in fibroblast or keratinocyte conditioned medium adhered poorly and remained rounded at 30 min. Immunodepletion of 804G conditioned medium with polyclonal antisera to laminin-5r, but not control antisera, abolished rapid cell spreading. Electron microscopy of HaCaT cells spread by incubation in 804G conditioned medium, but not control medium, revealed mature hemidesmosomes after 24 h. Rapid spreading was also observed in wells precoated with 804G conditioned medium or 804G cell-deposited matrix, but not with fibronectin, vitronectin, or laminin-1. Immunoblotting of 804G conditioned medium with anti-laminin-5r antibodies unveiled polypeptides of 150, 140, 135, and 100 kDa, identical by electrophoretic mobility to immunoreactive polypeptides in 804G deposited matrix. Our results suggest that addition of laminin-5r in a soluble form is sufficient to promote rapid spreading and hemidesmosome assembly in keratinocytes. The mechanism of soluble laminin-5r action may include efficient surface "priming" for cell adhesion. Soluble laminin-5r may have a physiologic role in morphogenesis and repair of the epidermis and may be of use for therapeutic applications.

A 105-kD lower lamina lucida antigen (p105) has been detected by autoantibodies (anti-p105) from patients with a novel immunobullous disease. To distinguish p105 from other known lamina lucida components, we performed comparative immunoblotting on purified human amniotic laminin-5 (kalinin), 804G matrix (enriched in laminin-5), and keratinocyte and fibroblast proteins using anti-804G matrix antibody (J-18) and anti-p105. J-18 labeled the truncated laminin-5 gamma 2 chain in amniotic laminin-5, 804G matrix, and keratinocyte conditioned medium, but did not label fibroblast cytosol. Conversely, anti-p105 did not label amniotic laminin-5 or 804G matrix, but did label p105 in both keratinocyte conditioned medium and fibroblast cytosol. J-18 labeled the 105-kD laminin-5 gamma 2 chain in reduced keratinocyte proteins and a 400-kD laminin-5 complex under non-reducing conditions. In contrast, anti-p105 labeled p105 under both reducing and non-reducing conditions but did not label a 400-kD protein complex. Similarly, comparative immunoblotting on keratinocyte proteins using anti-p105 and anti-laminin-1 revealed no commonly labeled protein bands. Electrophoretic fractionations by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of these fractions revealed that the peak fractions of keratinocyte proteins reactive with anti-p105 are different from those reactive with J-18. Furthermore, keratinocyte proteins fractionated by Mono Q anion-exchange chromatography revealed fractions immunoreactive with anti-p105, whereas J-18 showed no reactivity with these fractions. Two-dimensional gel electrophoresis and immunoblotting with anti-p105 revealed p105 to be an acidic protein with isoelectric points between 5.7 and 6.3, distinct from the isoelectric points of laminin-5 gamma 2 chain. We conclude that p105 is an acidic protein located in the lamina lucida and distinct from the truncated laminin-5 gamma 2 chain and the laminin-1 family.

Root surface cavities prepared in extracted premolars were restored with a selection of restorative materials. Prior to eosin dye immersion, one group of teeth was kept at constant temperature whilst another group underwent thermal cycling. The teeth were sectioned transversely through the restorations and an assessment of the degree of microleakage was used to compare the sealing ability of the selected materials. Eosin dye was able to discriminate more effectively between the microleakage behaviour of the restorative materials when samples were kept at a constant temperature.

An inter-district audit was carried out on the benefit to children of universal school entry medical examination, the appropriate use of professional time and to establish a baseline against which to measure future changes in services to under-fives and schoolchildren. The school health records of 1127 Cheshire schoolchildren, a random 10% sample, were scrutinised. There were significant district variations. Overall, 45% of children had problems not previously noted: 21% were serious enough to be referred; 11% could have had their problems managed by a well-trained school nurse but 9.8% would have required a medical examination for detection and assessment. These included conditions such as undescended testes, heart murmurs, squints and hernias and 8% required immediate action. The selection criteria, based on the absence of a three-year health check and/or parental or nurse concern, would have failed to identify 217 of the 491 children with new problems, some of which were serious. Targeting only an area of high deprivation or poor health would equally miss significant conditions. The inter-district variation of the numbers and diagnoses of new problems detected at school entry, and the difficulty in predicting which children will have important conditions, provide a good case for the continuation of the school entry medical examination for all children.

In the epidermis the autoantigen BP230 is a component of the hemidesmosomal plaque. We have developed a procedure for the isolation of BP230 from bovine tongue mucosa using chromatographic means. The identity of the isolated protein was confirmed by its recognition by bullous pemphigoid autoantibodies. A monoclonal antibody (MoAb230), generated against the purified protein, localizes to the region of the plaque of the hemidesmosome with which keratin bundles interact. Furthermore, the tissue distribution of BP230, assessed using MoAb230, suggests that BP230 or an immunologically related protein is a component of all hemidesmosomes. Ultrastructural analyses of the BP230 preparation reveal that the BP230 molecules assemble into macromolecular aggregates. The few images of individual intact molecules that we have observed in platinum replicas of rotary shadowed BP230 preparations suggest that BP230 is an elongate rod-shaped molecule. This is consistent with predictions based on the primary sequence of BP230 deduced from BP230 cDNAs reported by others. We discuss our results in relation to the potential function of BP230. Isolation of BP230 should now allow more rigorous biochemical analyses of potential protein-protein interactions of BP230 in the hemidesmosome.

Marginal leakage associated with a variety of restorative materials, placed in root surface cavities of extracted teeth, was assessed by immersing the teeth in either acidified gelatin, eosin dye, silver nitrate or a solution of radiocalcium. The allocation of a score, dependent on the depth of tracer penetration at the interface, allowed comparisons to be made between the leakage tests employed. The rank order of the four leakage tests was generally consistent. Eosin resulted in the most severe leakage, followed by silver nitrate and radiocalcium, whilst acidified gelatin was the least sensitive method for demonstrating marginal leakage. Differences in leakage scores were not, however, always statistically significant. None of the four tests investigated was ideal, although eosin dye was considered to be the most appropriate method of demonstrating marginal leakage associated with root restorations.

Using a serum sample of a bullous pemphigoid (BP) patient we have isolated a cDNA clone encoding a portion of a 180-kD polypeptide component of the hemidesmosome, the "BP180 autoantigen." The identity of the clone was confirmed by the generation of a fusion protein antibody that recognizes BP180 in both a basal epithelial cell extract of bovine tongue and extracts of human epidermal cells. Immunoelectron microscopy indicates that the 588-bp cDNA encodes a cytoplasmic fragment of BP180. Furthermore, the wide species reactivity of the fusion protein suggests that this portion of BP180 is highly conserved. In cultured human epidermal cells processed for confocal immunofluorescence microscopy, the fusion protein antibody generates a punctate cell substrate-associated staining pattern that is similar to that seen using BP230 antibodies. Using the original BP180 cDNA we have now isolated additional cDNA clones encoding approximately 1800bp of BP180 the 3' sequence of which overlaps with the sequence detailed in Guidice et al (J Clin Invest 87:734-738, 1991). Secondary structural analyses have been undertaken on the predicted amino acids encoded by the 1800bp. These suggest that the collagen-like sequences of BP180 described by Guidice et al (ibid.) are separated by a putative transmembrane region from the domain of BP180 recognized by our fusion protein antibody. Indeed, BP180 appears to belong to a relatively rare group of proteins in which the N-terminus is located in the cytoplasm and the C-terminus is extracellular. We detail some preliminary biochemical experiments in support of this hypothesis. We discuss possible functions of BP180 and BP230 in the hemidesmosome.

Extracellular matrix is considered to play an important role in determining the phenotype of cells with which it interacts. Here we have investigated the possibility that extracellular matrix is involved in specifying the pattern of keratin expression in epithelial cells. For these studies, we have developed an explant system in which epithelial cells from one type of stratified epithelial tissue, namely conjunctiva, are maintained on an extracellular matrix substrate derived from a different tissue, namely cornea. These ocular tissues are ideal for such analyses since they express distinct sets of keratins. For example, bovine conjunctival epithelium processed for immunofluorescence is not recognized by antibody preparations against keratin K3 or K12. In contrast, K3 and K12 antibodies generate intense staining in bovine corneal epithelium. At the immunochemical level, conjunctival cells in situ appear to possess no K12 and only trace amounts of K3, whereas corneal epithelial cells in situ possess both K3 and K12. When conjunctival cells are maintained on a corneal substrate with an intact basement membrane for 10 days in vitro they begin to express keratin K12 as determined by immunofluorescence. On the other hand, conjunctival cells that are maintained on a corneal substrate lacking a basement membrane fail to show staining with K12 antibodies. Conjunctival cells begin to show intense staining using K3 antibodies within about 10 days of being placed in culture regardless of their substrate. These results indicate that basement membrane can play a positive role in determining cell-specific expression of certain keratins such as K12. However, other keratins such as K3 may be "unmasked" and/or their expression may be upregulated simply by placing conjunctival epithelial cells in culture. We speculate that in conjunctiva K3 expression is influenced by certain negative exogenous factors. We discuss the possible means of regulation of keratin expression in our model system.

We have studied the effect of some specific enzyme inhibitors on DNA repair and replication after UV damage in Chinese hamster ovary cells. The DNA repair was studied at the level of the average, overall genome and also in the active dihydrofolate reductase gene. Replication was measured in the overall genome. We tested inhibitors of DNA polymerase alpha and delta (aphidicolin), of poly(ADPr) polymerase (3-aminobenzamide), of ribonucleotide reductase (hydroxyurea), of topoisomerase I (camptothecin), and of topoisomerase II (merbarone, VP-16). In addition, we tested the effect of the potential topoisomerase I activator, beta-lapachone. All of these compounds inhibited genome replication and all topoisomerase inhibitors affected the overall genome repair; beta-lapachone stimulated it. None of these compounds had any effect on the gene-specific repair.

In this review we will discuss the molecules involved in intermediate filament-desmosome and intermediate filament-hemidesmosome interactions, and the means by which certain of these molecules may bind to intermediate filaments. The possibility that intermediate filaments interact directly with peripheral membrane proteins and membrane lipids will also be addressed.

A patient with traumatic diaphragmatic hernia had his lesion present remotely after thoracoabdominal trauma as an asymptomatic chest wall mass. This is a rare and, perhaps, unique presentation of this entity.

A reproducible method of determining the porosity (pore volume) of cast investment materials is described. This procedure, requiring no highly specialized equipment, involved the infiltration of a test sample with paraffin wax and the weighing of the wax-infiltrated sample in both air and water. Pore-volume wax is then extracted with chloroform and the sample reweighed. The difference in weight is used to calculate the porosity of the investment material. Pore-volume data thus obtained are comparable to those resulting from standard techniques and far more sophisticated (and expensive) instrumentation. The porosities of hand-mixed samples of gypsum-bonded, phosphate-bonded and silica-bonded investments were 53.9 per cent, 36.9 per cent and 31.4 per cent and those of the vacuum-mixed samples were 52.5 per cent, 31.3 per cent and 26.9 per cent, respectively. The results show that gypsum-bonded investments were more porous than phosphate-bonded investments while silica-bonded investments were the least porous. Hand-mixed samples appeared to have greater porosity than vacuum-mixed samples.

Ten women with "idiopathic" edema had sodium and water retention and a rapid gain in weight when their accustomed intake of diuretics was suddenly stopped. The magnitude of these changes was directly related to the levels of plasma-rening activity before withdrawal of diuretics. Nine patients became edematous. Within 10 days of stopping diuretics, plasma-renin activity and urinary aldosterone excretion decreased to normal or below and within 20 days weight-gain and edema had subsided in seven patients. But the greatest gains in weight, in three patients, were sustained beyond 20 days. Nevertheless, a year later, two of these three patients were free of edema without the use of diuretics. Intermittent edema of unknown cause in most, if not all, otherwise healthy women seems to result from their use of diuretics, abetted in some patients by self-imposed flucuation of sodium and carbohydrate intake, and does not appear to be idiopathic.

We have undertaken an analysis of hemidesmosomes (HD) and their associated structures, intermediate filaments (IF) and anchoring fibrils (AF), in various types of basal cell carcinoma (BCC). Using a combination of electron microscopy and immunofluorescence microscopy we show that there is a correlation between the loss of HD and tumor type (i.e., in solid and infiltrative BCC hemidesmosomes are present, sometimes in reduced numbers), while there appears to be a lack of hemidesmosomes in cells of sclerosing specimens. Moreover, even though there is a loss of cytoplasmic constituents of the HD in sclerosing forms of BCC, this is not the case with regard to collagen VII, a component of AF, which are normally associated with the extracellular side of the HD. Collagen VII is localized to the basement membrane zone of tumor cells in the absence of the cytoplasmic constituents of HD. Furthermore, deposits of collagen VII occur in the connective tissue close to tumor cell populations in all but one of the BCC specimens we analyzed. In addition to modifications in HD and AF in BCC tissue, there are changes in the cytoskeletal elements of both tumor cells and the normal appearing epidermis that overlies tumor areas. In sclerosing BCC microfilaments are commonly observed along the basal portions of tumor cells where they abut the connective tissue. IF are often found interacting with these microfilaments. Indirect immunofluorescence analysis of tumor tissue using a monoclonal keratin antibody preparation, AE1, which in normal epidermis stains basal cells, reveals that AE1 antibodies only weakly stain tumor cells. Moreover, in the epidermis that overlies tumor cell regions AE1 antibodies stain suprabasal cells and not basal cells. This change in staining pattern generated by AE1 antibodies appears to depend upon the proximity of tumor cells. These results are discussed in relation to the organization of the HD and its associated AF and IF. The possibility that HD, IF, and AF antibody preparations may be of diagnostic use is raised.