Examining the prevalence of trans phantoms among transgender, nonbinary and gender diverse individuals: An exploratory study.

Background: Trans phantoms are bodily sensations of gendered body parts that a person was not born with (i.e., a phantom penis experienced by a trans man, or a phantom vagina experienced by a trans woman). This is distinct from the experience of many transgender and gender diverse (TGD) people, who experience awareness of their bodies as missing a gendered body part, or configuration, which is a major characteristic of gender dysphoria. Aims: Our purpose was to gain greater understanding of the prevalence and quality of trans phantoms. Methods: Data was gathered through a brief, online survey on trans embodiment. Respondents who had both completed the survey, and were deemed appropriate for inclusion in the study, based on their survey responses, comprised our sample of 1,446 adults. Results: Results indicated that trans phantoms are a typical embodied experience of TGD people. Almost 50% of study participants reported experiencing a trans phantom, most of whom also reported feeling erotic sensation in their phantom. Conclusions: Though the phenomenon of trans phantoms is not universal, it is clearly one that warrants further study.

Development of the Gender Embodiment Scale: Trans Masculine Spectrum.

Transgender and gender diverse (TGD) people have a variety of ways of embodying their gender. We present preliminary work on The Gender Embodiment Scale for trans masculine individuals as a collaborative product from a trans-identified community-engaged team. This scale provides researchers and clinicians a survey to diversify ways gender is understood and counteracts assumptions of a singular gender experience for TGD people. This scale reflects gender embodiment as individually unique and inclusive of the body, behavior, and social treatment. Use of the scale can enhance discussion and enable assessments regarding relative importance and satisfaction across items in these domains.

Potential limitations of transcription terminators used as transgene insulators in adenoviral vectors.

The presence of adenoviral cis-elements interfering with the activity of tissue-specific promoters has seriously impaired the use of transcriptional targeting adenoviruses for gene therapy purposes. As an approach to overcome this limitation, transcription terminators were previously employed in cultured cells to insulate a transgene promoter from viral activation. To extend these studies in vivo, we have injected into heart and skeletal muscle, adenoviruses containing the human growth hormone terminator and the cardiac-specific alpha-myosin heavy chain promoter (alphaMyHC) driving the chloramphenicol acetyltransferase (CAT) reporter gene. Promoterless CAT constructs were also tested to study interfering viral transcription and terminator activity. Here we demonstrate that the presence of a terminator can produce undesirable effects on the activity of heterologous promoters. Our analysis shows that in particular conditions, a terminator can reduce the tissue specificity of the transgene promoter. By RNAse protection assay performed on cardiac myocytes, we also show that adenoviral elements can direct high levels of autonomous transcription within the E1A enhancer region. This finding supports the model that passive readthrough of the transgene promoter is responsible for loss of selective expression.

Characterization of a replication-incompetent adenovirus type 5 mutant deleted for the preterminal protein gene.

An adenovirus type 5 mutant deleted for the preterminal protein (pTP) gene was constructed using cell lines that express pTP. The pTP deletion mutant virus is incapable of replicating in the absence of complementation and does not express detectable levels of viral mRNAs that are expressed only after the onset of replication. Accumulation of early-region mRNAs, including that for E1A, exhibits a lag relative to that observed from the wild-type virus. However, E1A mRNA accumulation attains a steady-state level similar to the level of expression during the early phase of infection with the wild-type virus. In 293-pTP cells (human embryonic kidney cells that express pTP in addition to high levels of adenovirus E1A and E1B proteins), the pTP deletion mutant virus replicates efficiently and yields infectious titers within 5-fold of that of the wild-type virus. The deletion of 1.2 kb of pTP-encoding sequence increases the size of foreign DNA that can be introduced into the virus and, with an absolute block to replication, makes this virus an important tool for gene therapy.

293 cell lines that inducibly express high levels of adenovirus type 5 precursor terminal protein.

293 cell lines that inducibly express high levels of adenovirus type 5 precursor terminal protein (pTP) under the control of a tetracycline-dependent promoter were constructed. To construct the cell lines expressing pTP, 293 cells were stably transfected with a plasmid encoding the tetracycline repressor/VP16 transactivator protein (tTA) using selection with hygromycin. Cell lines that expressed high levels of tTA activity were then stably transfected with plasmids in which pTP expression is directed by the tTA-dependent promoter from either a cDNA or a modified genomic construct using selection with G418. Cell lines that expressed high, inducible levels of pTP efficiently complemented a temperature-sensitive pTP mutant virus for growth and plaque formation at the nonpermissive temperature.

Non-nuclear oncogenes and the regulation of gene expression in transformed cells.

Oncogene products not localized to the nucleus regulate the expression of a diverse group of genes. The identities of genes regulated by non-nuclear oncogenes can supply insights into the changes at the cellular level that accompany the altered expression of such genes during the multi-step process of carcinogenesis. For example, one set of genes whose expression is affected by non-nuclear oncogenes are genes encoding extracellular proteases and components of the extracellular matrix. The expression of these genes during tumorigenesis could have important consequences for tumor invasiveness and metastasis. Genes regulated by non-nuclear oncogenes also define signal transduction pathways that allow communication between the plasma membrane and the nucleus. Oncogene-regulated nuclear targets provide a tool to approach the problem of cellular signal transduction and may contribute a clearer view of intracellular signaling pathways and the interactions between these pathways during cell growth and differentiation. Studying the regulation of these genes has revealed that c-jun and members of the ets family of transcription factors are important nuclear targets for the action of several non-nuclear oncogenes. This approach has also indicated that ras p21 is necessary for selective signal transduction events mediated by receptor and nonreceptor tyrosine kinases, including the colony-stimulating factor 1 (CSF-1) receptor, the product of the c-fms gene.

Mitogenic signaling by colony-stimulating factor 1 and ras is suppressed by the ets-2 DNA-binding domain and restored by myc overexpression.

The activity of p21ras is required for the proliferative response to colony-stimulating factor 1 (CSF-1), and signals transduced by both the CSF-1 receptor (CSF-1R) and p21ras stimulate transcription from promoter elements containing overlapping binding sites for Fos/Jun- and Ets-related proteins. A sequence encoding the DNA-binding domain and nuclear localization signal of human c-ets-2, which lacked portions of the c-ets-2 gene product necessary for trans activation, was fused to the bacterial lacZ gene and expressed from an actin promoter in NIH 3T3 cells expressing either the v-ras oncogene or human CSF-1R. Nuclear expression of the Ets-LacZ protein, confirmed by histochemical staining of beta-galactosidase, inhibited the activity of ras-responsive enhancer elements and suppressed morphologic transformation by v-ras as well as CSF-1R-dependent colony formation in semisolid medium. When CSF-1R-bearing cells expressing the Ets-LacZ protein were stimulated by CSF-1, induction of c-ets-2, c-jun, and c-fos ensued, but the c-myc response was impaired. Enforced expression of the c-myc gene overrode the suppressive effect of ets-lacZ and restored the ability of these cells to form colonies in response to CSF-1. NIH 3T3 cells engineered to express a CSF-1R (Phe-809) mutant similarly cannot form CSF-1-dependent colonies in semisolid medium and exhibit an impaired c-myc response, but expression of an exogenous myc gene resensitizes these cells to CSF-1 [M. F. Roussel, J. L. Cleveland, S. A. Shurtleff, and C. J. Sherr, Nature (London) 353:361-363, 1991]. The ability of these cells to respond to CSF-1 was also rescued by enforced expression of an endogenous c-ets-2 gene. The ets family of transcription factors therefore plays a central role in integrating both CSF-1R and ras-induced mitogenic signals and in modulating the myc response to CSF-1 stimulation.

An enhancer element responsive to ras and fms signaling pathways is composed of two distinct nuclear factor binding sites.

In order to precisely define the sequences that constitute the ras-responsive enhancers element present in the murine retrotransposon NVL3, point mutations were introduced into the previously defined minimal transcriptional enhancer DNA. Analyses of the effects of these point mutations in transient transfection experiments, in gel retention assays, and by methylation interference footprinting indicated that the enhancer element was composed of two binding sites for distinct nuclear factors. Both binding sites were required for activation of the enhancer by either ras or v-fms oncogenes, and the distinct nuclear factors were found in extracts from cells that contained either oncogene. UV cross-linking analysis revealed that the AP1-related binding site, TGACTCT, was recognized by a nuclear factor of apparent molecular size of 50 kilodaltons, that is probably c-jun. The other binding site, CAGGATAT, is very similar to sites recognized by the ets-family of transcription factors, and was recognized by the 120-kilodalton ras-responsive factor-1. Activation of the NVL3 element was reconstituted in an in vitro transcription assay. The ets-related binding site was necessary for this in vitro reconstitution of activity. Thus, the NVL3 enhancer is related to the previously described oncogene-responsive enhancer element present in polyoma virus and is also related to elements identified in several cellular genes known to be ras-responsive, including the transforming growth factor-beta 1 gene.

Negative regulation of transcription in vitro by a glucocorticoid response element is mediated by a trans-acting factor.

In vitro experiments with cell extracts prepared from a mouse mammary epithelial cell line demonstrated that a cis-acting glucocorticoid response element (GRE) of the mouse mammary tumor virus represses transcription from its homologous promoter. Competition transcription experiments, in which a molar excess of a restriction fragment that contains the GRE is added to the cell-free assay, revealed that a nuclear factor mediates in trans the negative regulation of mammary tumor virus transcription in vitro. Gel retention assays indicated that a factor in the extracts specifically recognizes the GRE. One unusual result of the gel retention studies was that heating the GRE probe to 65 degrees C before addition to a binding assay increases the formation of the specific protein-DNA complex 20-fold. Exonuclease III footprinting demonstrated that the sequences recognized by the factor are identical for either untreated or heat-treated probe. The footprinting also demonstrated that this factor recognizes sequences that are distinct from those recognized by the glucocorticoid receptor. A synthetic oligonucleotide based on the sequences identified by the footprinting experiments repressed the activity of a heterologous enhancer-promoter in vivo, as assayed by transient expression assays. We propose that this negative transcription element may control the basal level of expression of some glucocorticoid-modulated genes and may explain the insensitivity of certain tumor cells to steroid hormone action.

Recombinant vaccinia viruses carrying the N gene of human respiratory syncytial virus: studies of gene expression in cell culture and immune response in mice.

The construction and characterization of vaccinia virus recombinants carrying the nucleocapsid (N) protein gene of human respiratory syncytial (RS) virus are described. Recombinant viruses were constructed that contained the N gene oriented either positively or negatively with respect to the 7.5-kilodalton vaccinia virus promoter. In addition, a positively oriented recombinant was constructed that lacked an out-of-frame AUG codon in the 5'-terminal noncoding region. In HEp-2 cells, both positive-orientation recombinants induced the synthesis of a protein which comigrated with N protein and was precipitated by antisera to RS virus. Sera from mice immunized with these recombinants specifically precipitated the RS virus N protein. Analysis of mRNA and protein expressed from the recombinant N genes showed that deletion of the upstream AUG codon markedly improved the efficiency of protein synthesis. Mice were vaccinated with the high-expressing recombinant and subsequently challenged with live RS virus. The results of these experiments demonstrated that the immune response to N protein afforded a significant degree of protection against RS virus disease.

Correct sequence for the major nucleocapsid protein mRNA of respiratory syncytial virus.

A nucleotide sequence for the mRNA of the major nucleocapsid (N) protein gene of respiratory syncytial virus was reported previously (N. Elango and S. Venkatesen, 1983, Nucleic Acids Res. 11, 5941-5951). However, we have been unable to confirm part of this sequence as N mRNA-specific and suggest that the published sequence represents that of an aberrant chimeric transcript. Here we present an alternative sequence for the N mRNA and provide data supporting its authenticity. The corrected N mRNA sequence contains 1197 rather than 1427 nucleotides exclusive of poly(A), and encodes a protein of 391 rather than 467 amino acids. The calculated molecular weight for the 391-amino acid protein described by the sequence presented here is 42,600, in agreement with the molecular weight of 42,000 determined for the RS viral N protein by gel electrophoresis. In addition, we present sequence data from dicistronic RNAs that span the junction between the 1B protein and N cistrons, and the junction between the N and phosphoprotein (P) cistrons.

A liaison fellowship on a hemodialysis unit. A self-funded position.

The practice of liaison psychiatry has from its inception been hampered by an inadequate or non-existing funding base. A model is presented for funding an authentic liaison training program, fully supported by consultation-generated revenue. A specific description of the liaison teaching unit is given, which illustrates how the following objectives of the program were successfully met: provision of comprehensive biopsychosocial care, dissemination of psychological skills and knowledge to nonpsychiatrist staff, training of the liaison fellow, and generation of sufficient revenue to offset its costs.