Bullying perspectives among rural youth: a mixed methods approach.

INTRODUCTION: Few studies have examined violence among rural youth even though it is recognized as a societal concern. A mixed method, descriptive study was conducted to examine violence among rural youth including their perceptions and experiences of it. This article focuses specifically on the perceptions and experiences of bullying among rural youth that were generated from the Qualitative Phase One interviews and Quantitative Phase Two responses. METHOD: A mixed method study was conducted in two separate phases. The information generated from the Qualitative Phase One (n = 52) was used to develop a survey instrument employed in the subsequent Quantitative Phase Two (n = 180). The youth who were involved in each phase lived in different geographic areas of a Western Canadian province. The qualitative phase generated a number of comments about the experience of being bullied or how it felt to be a bully. In the survey instrument, specific questions related to bullying were embedded within it. Demographic information was collected in both phases of the study. Research assistants were used to collect the data in each phase. The transcripts from the qualitative phase were analyzed for categories and themes. The survey instrument included demographic questions and seventy questions that included a four-point Likert scale. The data were analyzed using SPSS v14 (SPSS Inc; Chicago, IL, USA). For this article, the survey questions that focused on bullying were considered alongside the qualitative comments in order to more fully understand the perceptions and viewpoints of rural youth regarding this particular aspect of violence. RESULTS: Conducting a mixed method study provides a more in-depth understanding of bullying among youth in the rural context. The pain and humiliation of being bullied provided a personalized understanding of the survey responses that indicated which youth are targets of bullying. For example, comments were made about being picked on because of personal characteristics such as being overweight or dressing in an unacceptable manner. In addition, bullies openly talked about the power they gained from their role. The frequency responses to the questions in the survey confirmed that bullies obtain power from their behavior and that youth who are different are bullied. The participants also noted that something needed to be done to address bullying but remarked that they would not seek professionals' help. DISCUSSION: The findings negate the myth that rural places are ideal places to raise children. Although the youth did not identify that they would access professionals, it is important for members of rural communities to acknowledge bullying, its impacts and how they can prevent it. Working from the social structure of rural communities is a first step in this process. CONCLUSION: Rural communities will benefit as a whole if bullying, an important societal concern, is addressed. Building on the social structure of rural communities is important, However, listening to rural youth themselves is the key if true change is to be implemented.

Holocene elephant seal distribution implies warmer-than-present climate in the Ross Sea.

We show that southern elephant seal (Mirounga leonina) colonies existed proximate to the Ross Ice Shelf during the Holocene, well south of their core sub-Antarctic breeding and molting grounds. We propose that this was due to warming (including a previously unrecognized period from approximately 1,100 to 2,300 (14)C yr B.P.) that decreased coastal sea ice and allowed penetration of warmer-than-present climate conditions into the Ross Embayment. If, as proposed in the literature, the ice shelf survived this period, it would have been exposed to environments substantially warmer than present.

Can professionals and nonprofessionals work together following a suicide?

Suicide is the taking of one's own life by one's own hand. It is often sudden and creates many emotional reactions for the survivors left behind. Survivor responses can be impacted by a range of circumstances, from how the person died to the reactions of people to the survivor. This contribution examines the uniqueness of survivor grief and how best to help survivors. It is suggested that their grief may be too quickly viewed as pathological, resulting in the premature medicalization of basically normal reactions. It is time for professionals and nonprofessionals to recognize boundaries and work toward mutual goals of health for those persons left behind after a suicide.

Field monitoring and performance evaluation of a field-scale in-well aeration system at a gasoline-contaminated site.

Several in-well aeration (IWA) technologies have been used since the early 1990s, but few field studies have been performed to evaluate the extent of water circulation around IWA systems. In this study, 27 discrete monitoring points (MPs) were installed at a gasoline-contaminated site to assess the efficacy of IWA. Pressure transducers and dissolved oxygen (DO) probes were sealed into the MPs, allowing them to be used to characterize subsurface changes in total head and DO with depth, distance and orientation from a central injection well. No change in DO or in hydrocarbon total mass or distribution occurred across the site during two trials (41 and 20 days) of the system. Water level fluctuations during the trials were similar in all MPs, and were due to seasonal water table changes and rainfall events. No circulation cell was established around the IWA well after 41 days of operation, and the impact of the well extended less than 90cm from it. Groundwater only circulated through the sand pack around the well. Little, if any, recharge occurred through the lower screen. Silt accumulated in the well, limiting its operation time, even with a fabric filter sock over the lower screen. Obviously, IWA was ineffective at this site, probably because the horizontal hydraulic conductivity (K(h)) of the soil opposite the lower screen was low (0.09cm per day) and because the distance between the two screens was short relative to the borehole radius. Long remediation times would likely make IWA unattractive at this or other sites where the K(h) of the soil is so low that the air injection rate would have to be low to prevent blowing the well dry.

Field monitoring and performance evaluation of an in situ air sparging system at a gasoline-contaminated site.

In situ air sparging (IAS) has been used since the mid-1980s, but few carefully designed field studies have been performed to evaluate its effectiveness. In this study, 27 discrete monitoring points (MPs) were installed at a gasoline-contaminated site to investigate the efficacy of IAS. Each MP was instrumented with a pressure transducer and a Technalithics dissolved oxygen (DO) probe, and located so they could be used to characterize subsurface changes in total head and DO with depth, distance and orientation around a central injection well. Because the blower over-heated and automatically shut down after approximately 30 min and short-circuiting of air into two MPs occurred within 2 min, the study was designed as three sets of three 30-min trials. Longer trials would not have yielded different nor more insightful results. A volume of soil was not oxygenated during any injection. Instead, air traveled directly to at least four of seven different MPs during eight of the nine trials, probably as a result of an air bubble forming beneath a confining layer. The order of air arrival at the MPs varied during the first few trials, but once a preferential pathway was established, it did not collapse between trials and provided the shortest distance to the vadose zone during subsequent trials. Oxygen uptake rates estimated for MPs that received air during any trial exceeded the consumption rates of the Technalithics DO probes, and indicate that the probes could be used for estimating oxygen transfer during system operation or for oxygen uptake measurements during shut-down tests. The data from the monitoring system indicate that IAS is infeasible for remediation of soil and groundwater at this site due to its low horizontal hydraulic conductivity. Similar behavior is anticipated when IAS is applied at other sites with low hydraulic conductivity materials.

DHR3 is required for the prepupal-pupal transition and differentiation of adult structures during Drosophila metamorphosis.

Pulses of the steroid hormone ecdysone activate genetic regulatory hierarchies that coordinate the developmental changes associated with Drosophila metamorphosis. A high-titer ecdysone pulse at the end of larval development triggers puparium formation and induces expression of the DHR3 orphan nuclear receptor. Here we use both a heat-inducible DHR3 rescue construct and clonal analysis to define DHR3 functions during metamorphosis. Clonal analysis reveals requirements for DHR3 in the development of adult bristles, wings, and cuticle, and no apparent function in eye or leg development. DHR3 mutants rescued to the third larval instar also reveal essential functions during the onset of metamorphosis, leading to lethality during prepupal and early pupal stages. The phenotypes associated with these lethal phases are consistent with the effects of DHR3 mutations on ecdysone-regulated gene expression. Although DHR3 has been shown to be sufficient for early gene repression at puparium formation, it is not necessary for this response, indicating that other negative regulators may contribute to this pathway. In contrast, DHR3 is required for maximal expression of the midprepupal regulatory genes, EcR, E74B, and betaFTZ-1. Reductions in EcR and betaFTZ-F1 expression, in turn, lead to submaximal early gene induction in response to the prepupal ecdysone pulse and corresponding defects in adult head eversion and salivary gland cell death. These studies demonstrate that DHR3 is an essential regulator of the betaFTZ-F1 midprepupal competence factor, providing a functional link between the late larval and prepupal responses to ecdysone. Induction of DHR3 in early prepupae ensures that responses to the prepupal ecdysone pulse will be distinct from responses to the late larval pulse and thus that the animal progresses in an appropriate manner through the early stages of metamorphosis.

A novel tumor-specific human single-chain Fv selected from an active specific immunotherapy phage display library.

A colon tumor-associated antigen, CTAA 28A32-32K (CTA # 2E), related to the annexin family of proteins, was initially identified by its reactivity with a low affinity human IgM monoclonal antibody (mAb), 28A32. Both in vitro lymphoproliferative assays with human peripheral blood lymphocytes and delayed type hypersensitivity responses in patients immunized with autologous colon tumor cells indicated that CTA # 2E elicits potent T cell mediated responses and may be an important antigen in the development of a generic colorectal vaccine (Pomato et al. Vaccine Res 1994;3:145-161). A CTA # 2E-specific, murine hybridoma-derived mAb, 5-11A, which recognizes the amino-terminus of the tumor-associated antigen, exhibited qualitative human colon tumor-specific immunohistochemical reactivity. To rapidly develop a human mAb with similar antigen specificity and tumor reactivity as the murine 5-11A mAb, antibody phage display technology was employed. Two human antibody phage display libraries with 3.1 x 10(7) and 2.3 x 10(8) members were prepared from the variable region genes expressed by circulating B cells of patients undergoing active specific immunotherapy (ASI) with autologous tumor cells, predominantly from the colon, admixed with Bacille Calmette-Guerin (BCG). A CTA # 2E-reactive human single-chain (sc)Fv was selected by panning the larger library on decreasing concentrations of biotinylated tumor-associated antigen in solution. It exhibited similar antigen specificity as the murine hybridoma-derived 5-11A scFv, requiring the presence of the CTA # 2E amino-terminus for reactivity. This human scFv exhibited qualitative human colon tumor-specific immunohistochemical reactivity when displayed as a gene III fusion protein on phage. When reconstructed and expressed as an intact human IgG1, K mAb, its qualitative colon tumor-specificity was unaltered. Two other CTA # 2E-reactive human scFvs were selected from the smaller library by panning initially on decreasing concentrations of CTA # 2E coated to polystyrene and then on biotinylated CTA # 2E in solution. These human scFvs, which exhibited modest reactivity with different epitopes on the CTA # 2E antigen, did not exhibit human colon tumor-specific immunohistochemical reactivity.

The RXR homolog ultraspiracle is an essential component of the Drosophila ecdysone receptor.

Pulses of the steroid hormone ecdysone function as key temporal signals during insect development, coordinating the major postembryonic developmental transitions, including molting and metamorphosis. In vitro studies have demonstrated that the EcR ecdysone receptor requires an RXR heterodimer partner for its activity, encoded by the ultraspiracle (usp) locus. We show here that usp exerts no apparent function in mid-third instar larvae, when a regulatory hierarchy prepares the animal for the onset of metamorphosis. Rather, usp is required in late third instar larvae for appropriate developmental and transcriptional responses to the ecdysone pulse that triggers puparium formation. The imaginal discs in usp mutants begin to evert but do not elongate or differentiate, the larval midgut and salivary glands fail to undergo programmed cell death and the adult midgut fails to form. Consistent with these developmental phenotypes, usp mutants show pleiotropic defects in ecdysone-regulated gene expression at the larval-prepupal transition. usp mutants also recapitulate aspects of a larval molt at puparium formation, forming a supernumerary cuticle. These observations indicate that usp is required for ecdysone receptor activity in vivo, demonstrate that the EcR/USP heterodimer functions in a stage-specific manner during the onset of metamorphosis and implicate a role for usp in the decision to molt or pupariate in response to ecdysone pulses during larval development.

The 3'-untranslated region of membrane exon 2 from the gamma 2a immunoglobulin gene contributes to efficient transcription termination.

Elements of the mouse Immunoglobulin gamma 2a gene, near the membrane-specific poly(A) addition site, were inserted into a heterologous location in either a synthetic mouse gamma 2b gene or a gpt/SV40 chimeric gene and then assayed for their ability to terminate RNA polymerase II transcription in isolated nuclei of transfected myeloma cells. The intact gamma 2a membrane-specific 3'-untranslated region, with its potential stem loop forming sequences and poly(A) site, is able to efficiently terminate transcription in the absence of the downstream region in which transcription normally terminates (term). Termination efficiency in the presence of the termination fragment decreases either when sequences specifying a potential stem/loop, upstream of the poly(A) region, are interrupted or when the stronger membrane poly(A) site is substituted with a weaker, secretory-specific poly(A) site. We therefore conclude that the gamma 2a membrane-specific untranslated region plays a major role in specifying downstream termination. We further conclude that the immunoglobulin gamma 2a, membrane-specific, 3'-untranslated region can function in the context of the gpt gene, driven by an SV40 promoter, to terminate transcription in a poly(A) site dependent fashion.

Establishment, molecular rescue, and expression of 123AV16-1, a tumor-reactive human monoclonal antibody.

The human monoclonal antibody (mAb) 123AV16-1 was generated by Epstein-Barr virus transformation of peripheral blood lymphocytes from a colorectal patient undergoing active specific immunotherapy with an autologous tumor cell-Bacille Calmette-Guérin vaccine. Direct immunohistochemical staining of tumor and normal pairs of tissues indicated that this human IgA1, lambda 2 mAb preferentially reacted with colon tumor epithelium. To generate a recombinant derivative of this Epstein-Barr virus-transformed cell line, we isolated the expressed complete heavy and light chain genes by a novel strategy and cloned them into modified pSV2-neo and pSV2-gpt expression vectors. The recombinant 123AV16-1 human mAb was expressed in both a murine myeloma and a human-murine heteromyeloma and was secreted as both monomers and dimers. The recombinant 123AV16-1 mAb expressed by both cell lines reacted with human colon tumor xenografts in a manner similar to the mAb derived from the Epstein-Barr virus-transformed cell line, indicating that the antibody specificity was not appreciably altered during the molecular rescue, cloning, or expression.

Baboon and cotton-top tamarin B2m cDNA sequences and the evolution of primate beta 2-microglobulin.

Nonhuman primates represent phylogenetic intermediates for studying the divergence of human and murine beta 2Ms. We report the nucleotide sequences of B2m cDNA clones from a baboon cell line, 26CB-1 (Papio hamadryas; primates: Cercopithecoidea), and a cotton-top tamarin cell line, 1605L (Saguinus oedipus; primates: Ceboidea). The baboon and tamarin B2m sequences indicate a very slow rate of B2m evolution in primates relative to that in murid rodents. Phenotypic evolution of beta 2M has also been very conservative in primates, with only 9-14 substitutions separating baboon or tamarin beta 2Ms from those of humans or orangutans. Analyses of silent and amino-acid-altering nucleotide substitutions provide evidence that negative selection has acted to limit variability in beta strands of primate beta 2Ms, while positive selection has promoted diversity in non-beta-strand regions of murine beta 2Ms. No evidence for the action of selection upon beta 2M residues that contact the class I heavy chain was found in primates or mice. The finding that different selective forces have operated upon primate and murine beta 2Ms suggests that beta 2M may have evolved to serve distinct functions in primates and mice.

v-erbA and citral reduce the teratogenic effects of all-trans retinoic acid and retinol, respectively, in Xenopus embryogenesis.

Treatment of late blastula/early gastrula stage Xenopus embryos with all-trans retinoic acid results in disruption of the primary body axis through effects on both mesoderm and neuroectoderm. This effect of retinoic acid, coupled with the known presence of retinoic acid in Xenopus embryos has led to the proposal that retinoic acid may be an endogenous morphogen providing positional information in early development. To further elucidate the role of retinoic acid in early Xenopus development, we have attempted to interfere with the retinoic acid signalling pathway both at the level of retinoic acid formation, by treatment with citral (3,7-dimethy-2,6-octadienal), and at the level of nuclear retinoic acid receptor function, by microinjection of v-erbA mRNA. The feasibility of this approach was demonstrated by the ability of citral treatment and v-erbA mRNA injection to reduce the teratogenic effects of exogenous retinol and retinoic acid, respectively, in early Xenopus development. Interestingly, v-erbA mRNA injection and citral treatment of gastrula stage embryos resulted in tadpoles with a similar set of developmental defects. The defects were chiefly found in tissues that received a contribution of cells from the neural crest, suggesting that at least a subset of neural crest cells may be sensitive to the endogenous level of retinoic acid. In accord with this proposal, it was found that the expression patterns of two early markers of cranial neural crest cells, Xtwi and XAP-2, were altered in embryos injected with v-erbA mRNA. These results indicate that structures in addition to the primary axis are regulated by retinoic acid signalling during early Xenopus development.

Reconstitution of retinoid X receptor function and combinatorial regulation of other nuclear hormone receptors in the yeast Saccharomyces cerevisiae.

The nuclear hormone receptor family of transcription factors regulates gene expression via a complex combinatorial network of interactions. Of particular interest is the ability of retinoid X receptors (RXRs) to form heterodimers with retinoic acid receptors (RARs) and thyroid hormone receptors (TRs), thereby modifying their activities. We report here that RXR, RAR, and TR function can be reconstituted in the yeast Saccharomyces cerevisiae and demonstrate that the combinatorial regulation seen in vertebrate cells can be reproduced in the yeast background. Using this system, we have shown that RARs respond to a wide variety of retinoid ligands but that RXRs are specific for the 9-cis isomer of retinoic acid. RXR enhanced the activity of RARs and TRs on a variety of hormone response elements without demonstrably altering their DNA specificity. Interestingly, the ability of RXR to potentiate gene activation by RARs and by TRs varied for different receptor isoforms.

Variables affecting the T cell receptor V beta repertoire heterogeneity of T cells infiltrating human renal allografts.

Donor-specific, alloreactive T cell lines may be grown from cells infiltrating human renal allografts. These T cell lines utilize restricted T cell receptor (TCR) beta-chain variable (V beta) gene repertoires, although long-term culture appears to be necessary for restriction to be observed. This study was undertaken to determine the effects of potential selective pressures on the TCR repertoires of allograft-infiltrating cells. TCR V beta repertoires of 30 allograft-derived T cell populations, cultured for defined, short time periods, were examined using polymerase chain reaction. When first derived, V beta repertoires of graft-infiltrating T cells were as heterogeneous as those of peripheral blood lymphocytes (PBL). There was no relationship between the length of time an allograft was in situ or the extent of HLA mismatch and repertoire heterogeneity. Repertoire restriction was positively correlated with the length of time cells were cultured in vitro. Long-term, alloreactive mixed lymphocyte reactions (MLR), established from normal, unsensitized PBL, also demonstrated V beta repertoire restriction during expansion in vitro. Restricted alloreactive populations emerged much more slowly from the MLR than from the allograft-derived cultures, however, implying that graft infiltrates contain previously activated populations of T cells. This observation, taken together with the fact that long-term, graft-derived cell lines maintain donor specificity, suggests that functional subsets must be allowed to emerge from heterogeneous infiltrates before TCR repertoire may be correlated with alloreactivity and/or graft rejection.

A single amino acid mutation in CDR3 of the 3-14-9 L chain abolished expression of the IDA 10-defined idiotope and antigen binding.

The ABPC 48 myeloma protein and the 3-14-9 mAb derive their V region genes from the same VH and V kappa gene families. They also share a cross-reactive idiotope defined by the anti-Id mAb IDA 10. Whereas ABPC 48 is specific for bacterial levan, 3-14-9 showed a significant Ag-binding activity to aminophenyl-beta-N-acetylglucosaminide (AZO). In order to define the molecular basis of idiotope expression and Ag-binding activity, we have cloned the genes encoding the 3-14-9 H and L chain V region genes, generated antibodies that carry mutations within the L chain genes, by site-directed mutagenesis, and investigated the effects of those mutations with respect to IDA 10 idiotope expression and binding to AZO. Our findings show that, whereas expression of the IDA 10-defined idiotope requires association of both the H and L chains, a single change (glycine to phenylalanine) at position 91 in the third complementarity-determining region of the L chain abolished both idiotope expression and Ag-binding activity. In addition, a L chain change of alanine to threonine at position 25 allowed idiotope expression to some extent but significantly reduced binding activity to AZO. These data suggest that a single amino acid change can play a crucial role in the functional activity and structural integrity of antibodies.

T cell receptor V beta gene usage in HLA-DR1-reactive human T cell populations. The predominance of V beta 8.

The functional specificity and T cell receptor (TCR) V beta gene expression of three class II HLA-DR1-reactive human T cell populations were examined. WP2, a renal allograft-derived, long-term CD4+ T cell line, was specifically cytotoxic for DR1, one of the mismatched antigens present on the allograft. Initial studies of WP2 using six TCR V beta-specific mAb revealed a predominance of T cells expressing a member of the V beta 8 gene family. A smaller, yet significant, number of cells expressed TCR using V beta 5.1. Semiquantitative V beta-specific polymerase chain reaction (PCR) analyses of RNA derived from this T cell line confirmed the presence of V beta 8 and V beta 5.1 messages. The PCR signal for V beta 8 was the strongest, followed by those for V beta 4 and V beta 5. An earlier WP2 culture had a very similar PCR profile, with a dominant signal for V beta 8, although signals for V beta 4 and V beta 5 were considerably lower. Previous PCR analyses of eight other renal allograft-derived, long-term T cell lines, grown under identical in vitro conditions, revealed no other example of predominant usage of V beta 8. We established two replicate long-term, anti-DR1 mixed lymphocyte reactions using PBL from two unrelated normal donors as responder and stimulator. The MLR were given alloantigen every 10 days, and RNA was obtained from the cultured cells immediately prior to each stimulation. PCR analyses of RNA taken at 10-day intervals over a total of 60-70 days indicated that, although the MLR were initially quite heterogeneous with regard to V beta message expression, by the end of the fourth or fifth Ag cycle the predominant PCR signals observed in both MLR were for V beta 8. These results suggest that T cells using V beta 8 gene-encoded segments as part of their TCR may have a selective advantage in responses to DR1.

PCR-based analysis of the T-cell receptor V beta multigene family: experimental parameters affecting its validity.

The validity of semiquantitative, PCR-based analysis of gene expression within a multigene family, the human T-cell receptor (TCR) beta chain variable region family, was investigated. Primer comparability was addressed by grouping hybridization temperatures and limiting the size range of amplified fragments. Primers selected satisfied criteria for comprehensiveness, match to targets and discrimination of nontargets. Specificity was enhanced by maximizing mismatches with nontargets and using an elevated hybridization temperature. Reaction conditions are described that ensure specificity while maintaining sensitivity. Several results confirmed primer specificity. Limits on precision were documented: probable error was 3%-7% of mean value for target prevalences (% of all TCR mRNA represented by a particular V beta) in the 5%-40% range. Accuracy was limited by the nonlinear relationship between target prevalence and signal obtained. Because of this relationship, the effect of the observed limits of precision varied. Valid distinctions were possible between sufficiently separated prevalences, i.e., 0%-1%, 3%-5%, 10%, 30%, greater than 50%. Additional concerns addressed include: standardization of signals, coamplifiation and effects of primer artifacts, and the nature of the mRNA pool. Only when theoretical and practical limits in precision and accuracy are acknowledged can semiquantitative, PCR-based analysis be used with confidence to assess gene usage within a large, multigene family.

T cell receptor V beta gene usage in allograft-derived cell lines analyzed by a polymerase chain reaction technique.

T cell receptor V beta gene usage in renal allograft-derived T cell lines was investigated using a semiquantitative, polymerase chain reaction (PCR)-based technique. A panel of V beta-specific primers was carefully designed to have uniform hybridization characteristics, enhancing comparability, and high specificity based on 3' mismatches between primers and nontarget V regions. PCR data were in concordance with previous phenotypic, functional, and Southern blot analyses, but the PCR technique proved to be more sensitive and comprehensive. For example, the EH3 cell line was confirmed to be polyclonal, with a number of different V beta genes mediating anti-DR8-linked reactivity. Sequence analysis showed the most prevalent signals obtained by PCR, those for V beta 7 and V beta 20, probably corresponded to the only two rearrangements detected by Southern. The J2 cell line was shown to be polyclonal in early culture, when it was mostly CD8+ and B35-reactive. Late cultures and a subclone were CD4+, possessed DR3-linked reactivity, and evidenced only one PCR signal: V beta 6. Similarly, the MH3 cell line was shown to be polyclonal early in culture, when it was reactive toward A11 and A29. In late cultures, when Southern analysis indicated clonality, only anti-A11 reactivity was maintained, and the predominant PCR signal was for V beta 17. Other alloreactivities were also attributed to specific V beta. The Kng cell line, alloreactive to A28, yielded two strong PCR signals; V beta 2 and V beta 9. The Mijo line, DR1-reactive, gave a predominant signal for V beta 12. Thus in spite of variable in vitro selection occurring in some lines and great initial heterogeneity documented by PCR, this technique was still capable of identifying V beta genes that persist in vitro, become predominant, and are associated with specific, allograft-directed alloreactivities.

Functional interaction between the two zinc finger domains of the v-erb A oncoprotein.

The v-erb A oncogene of avian erythroblastosis virus is a mutated and virally transduced copy of a host cell gene encoding a thyroid hormone receptor. The protein expressed by the v-erb A oncogene binds to DNA and acts as a dominant negative inhibitor of both the thyroid hormone receptor and the closely related retinoic acid receptor. The v-erb A protein has sustained two amino acid alterations within its DNA-binding domain relative to that of c-erb A, one of which, at serine 61, is known to be important for v-erb A function in the neoplastic cell. We report here that the second alteration, at threonine 78, also plays an important, although more indirect, role: alteration of the sequence at threonine 78 such that it resembles that of c-erb A can act as an intragenic suppressor and can partially restore function to a v-erb A protein rendered defective due to a mutation at position 61. Threonine 78 lies within the D-box of the v-erb A protein, a region thought to mediate receptor-receptor dimerizations, and is not in physical proximity to the serine at position 61. It therefore appears that an indirect interaction occurs between these two sites and that this interaction is crucial for v-erb A function.