Radiographic Differentiation of Cranial Mediastinal Lymphomas from Thymic Epithelial Tumors in Dogs and Cats.

In both dogs and cats, the most common cranial mediastinal masses (CMMs) are lymphoma and thymic epithelial tumors (TETs). Pretreatment differentiation of these tumors using fine needle aspiration or biopsy is essential because lymphomas are treated medically, whereas TETs are treated surgically. The purpose of this retrospective study was to determine whether thoracic radiographic findings can be used to aid clinicians in preliminarily differentiating the two tumor types before cytology or histopathology results become available. Medical records, available cytologic or histologic samples, and thoracic radiographs were evaluated for 62 dogs and 28 cats. Seventeen radiographic criteria were assessed by two examiners, and regression modeling was performed to test for significant predictors of tumor type. In dogs, CMMs with at least two well-defined radiographic margins on a lateral view and CMMs causing a rightward shift of the cardiac silhouette on a ventrodorsal or dorsoventral view were significantly more likely to be TETs than lymphomas (P < .001 and P < .01, respectively). No significant predictive variables were identified in cats. Radiographic findings do not eliminate the need for invasive sampling, but in dogs, they may guide the clinician in providing preliminary information to owners regarding the staging and therapeutic measures that may eventually be recommended.

ATF4 is a mediator of the nutrient-sensing response pathway that activates the human asparagine synthetase gene.

Transcription from the asparagine synthetase (A.S.) gene is increased in response to either amino acid (amino acid response) or glucose (endoplasmic reticulum stress response) deprivation. These two independent pathways converge on the same set of genomic cis-elements within the A.S. promoter referred to as nutrient-sensing response elements (NSRE) 1 and 2, both of which are necessary for gene activation. The NSRE-1 sequence was used to screen ATF/CREB family members by electrophoresis mobility shift assays and supershift by specific antibodies. The results indicated that ATF4 binds to the NSRE-1 sequence and that the amount of the ATF4 complex was increased when extracts from amino acid-deprived or glucose-deprived cells were tested. Using electrophoresis mobility shift assay experiments and a probe that contained both NSRE-1 and NSRE-2, mutation of the NSRE-1 sequence completely prevented formation of the ATF4-containing complexes, whereas mutation of the NSRE-2 sequence did not. Overexpression of ATF4 increased A.S. promoter-driven transcription, whereas an inhibitory dominant negative ATF4 mutant blocked both basal and starvation-enhanced transcription. Collectively, the results provide both in vitro and in vivo evidence for a role of ATF4 in the transcriptional activation of the A.S. gene in response to nutrient deprivation.

The mechanism for transcriptional activation of the human ATA2 transporter gene by amino acid deprivation is different than that for asparagine synthetase.

After amino acid deprivation, the mRNA content for both asparagine synthetase (AS) and the system A transporter ATA2 is increased. The purpose of the reported experiments was to characterize the molecular mechanism for the ATA2 gene and to contrast the ATA2 regulatory characteristics with those of AS. Amino acid limitation was initiated by incubation of HepG2 human hepatoma cells in either amino acid-free Krebs-Ringer bicarbonate buffer or culture medium lacking the single amino acid histidine. For ATA2, like AS, the elevated mRNA content was due to increased transcription. However, there were fundamental differences between the mechanisms for nutrient regulation of the AS and ATA2 genes. When cells were deprived of amino acids, there was a lag period of approximately 4 h before an increase in AS mRNA occurred, whereas the elevation of ATA2 mRNA was readily detectable at 2-4 h. Consistent with these observations, de novo protein synthesis was absolutely required for the activation of the AS gene, but the increase in ATA2 mRNA was largely independent of protein synthesis. Furthermore, in contrast to AS, transcription from the ATA2 gene was not increased by glucose deprivation. Given this lack of ATA2 transcriptional activation by glucose starvation and that the induction of the AS gene by glucose or amino acid starvation is mediated by common genomic elements, it is likely that the ATA2 gene does not contain the same genomic amino acid-responsive cis-elements as the AS gene.