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Concomitant rupture of the subclavian vessels and the left main bronchus caused by blunt trauma is a serious condition. Moreover, the diagnosis of a tracheobronchial injury with rupture of the subclavian vessels can be difficult. This report describes the case of a 33-year-old man who suffered from blunt trauma that resulted in the rupture of the left subclavian artery and vein. The patient underwent an operation for vascular control. On postoperative day 3, the left main bronchus was found to be transected on a computed tomography scan and bronchoscopy. The transected bronchus was anastomosed in an end-to-end fashion. He recovered without any notable problems. Although the bronchial injury was not detected early, this case of concomitant rupture of the great vessels and the airway was successfully treated after applying extracorporeal membrane oxygenation.

Background: Re-expansion pulmonary edema (REPE) is a lethal complication that can occur after inserting a chest tube. However, no clinical research reports have analyzed the progress and treatment of REPE, except for a single case report review. We retrospectively analyzed the common clinical characteristics, clinical progress, and treatment outcome of REPE with respiratory failure. Methods: We retrospectively reviewed the clinical features, treatment, and outcomes of eight patients with REPE who required ventilator care from March 2004 to March 2018. Results: The mean PaO 2 /FiO 2 ratio of the patients immediately after intubation was 106.5±20.2 (range, 75–128), which indicated severe hypoxia in all cases. On the first day of treatment, most of the patients showed improvement in hypoxia symptoms, and most improved to the extubation level on day 3. The mean duration of ventilator use was 2.5±0.8 days (range, 1–4 days), and the mean duration of inotropic drug use was 1.1±0.7 days (range, 0–2 days). The mean intensive care unit (ICU) stay was 4.4±1.5 days (range, 3–8 days). No deaths were recorded during the study period, and no cases of permanent complications due to REPE were observed. Conclusions: REPE requiring ventilator care is a lethal disease. However, continued development of ICU care has dramatically decreased mortality. Low positive end-expiratory pressure (PEEP), low tidal and high O 2 ventilator care represents the most appropriate treatment for REPE.

Purpose: Tracheobronchial injuries caused by trauma are rare, but can be life threatening. The objective of this study was to evaluate the surgical outcome for patients with tracheobronchial injuries and to determine the difference, if any, between the outcomes for patients with penetrating trauma and those for patients with blunt trauma. Methods: From January 2010 to June 2015, 40 patients underwent tracheobronchial repair surgery due to trauma. We excluded 14 patients with iatrogenic injuries, and divided the remaining 26 into two groups. Results: In the blunt trauma group, injury mechanisms were motor vehicle accident (9 cases), free falls (3 cases), flat falls (1 case) and mechanical injury (1 case). In the penetrating trauma group, injury mechanisms were stab wounds (10 cases), a gunshot wound (1 case) and a stab wound caused by metal pieces (1 case). The mean RTS (Revised Trauma Score) was 6.89±1.59 (range: 2.40-7.84) and the mean ISS (Injury Severity Score) was 24.36±7.16 (range: 11-34) in the blunt group; the mean RTS was 7.56±0.41 (range: 7.11-7.84), and the mean ISS was 13±5.26 (range: 9-25) in the penetrating trauma group. In the blunt trauma group, 9 primary repairs, 1 resection with end-end anastomosis, 2 lobectomies, 1 sleeve bronchial resection and 1 pneumonectomy were performed. In the penetrating trauma group, 10 primary repairs and 2 resections with end-end anastomosis were performed. Complications associated with surgery were found in one patient in the blunt trauma group, and one patient in the penetrating trauma group. No mortalities occurred in either groups. Conclusion: Surgical management of a traumatic tracheobronchial injury is a safe procedure for both patients with a penetrating trauma and those with a blunt trauma. [ J Trauma Inj 2016; 29: 1-7 ]

【Penetrating neck trauma involving spinal cord injury is relatively uncommon, but can be life-threatening. We report a case of 59-year-old female who presented with hypotension after stab injury self-inflicted with a scissor to her neck. Although Open removal of the scissor and control of bleeding were successfully done, penetration of spinal cord resulted in a neurologic impairment.】

Vacuum-assisted closure therapy is an alternative method for a massive subcutaneous emphysema treatment. It is easily applicable and shows rapid effectiveness in massive subcutaneous emphysema, intractable with chest tube drainage.

There are various methods for approaching the aortic arch, such as median sternotomy or lateral thoracotomy. However, accessing the site of distal anastomosis is problematic when the distal arch is extensively involved. We report a case of extended aortic arch replacement and coronary artery bypass through the L-incision approach.

Chest drainage systems are usually composed of chest tube and underwater-seal bottle. But this conventional system may restrict patients doing exercise and give clinicians obscure data about when to remove tubes because there is no objective indicator. Recently developed digital chest drainage systems may facilitate interpretation of the grade of air leak and make it easy for clinicians to decide when to remove chest tubes. In addition, with combination of wireless internet devices, monitoring and managing of drainage system distant from the patient is possible.Sixty patients of primary pneumothorax were included in a prospective randomized study and divided into two groups. Group I (study) consisted of digital chest drainage system while in group II (control), conventional underwater-seal chest bottle system was used. Data was collected from January, 2012 to September, 2013 in Eulji University Hospital, Daejeon, Korea.There was no difference in age, sex, smoking history and postoperative pain between two groups. But the average length of drainage was 2.2 days in group I and 3.1 days in group II (P0.006). And more, about 90% of the patients in group I was satisfied with using new device for convenience.Digital system was beneficial on reducing the length of tube drainage by real time monitoring. It also had advantage in portability, loudness and gave more satisfaction than conventional system. Moreover, internet based digital drainage system will be a good method in thoracic telemedicine area in the near future.

Purpose Endoscopic thoracic sympathetic surgery is effective for treating palmar hyperhidrosis, although compensatory sweating (CS) is a significant and annoying side effect. The purpose of this study was to compare the results of limited resection at two different locations. Methods From May 2004 to June 2009, T3 sympathicotomy (group I) was performed in 46 patients and T3,4 ramicotomy (group II) was performed in 43 patients during the same period. T3 sympathicotomy (group I) and T3,4 ramicotomy (group II) were performed during the same period. Using questionnaires, completed by the patients, the satisfaction rates and grades of CS were analyzed. Results No significant differences in age distribution or sex ratios were observed between the two groups. The satisfaction rate was 91.3 % in group I and 79.1 % in group II. The operation time was 19.8 (±6.6) min in group I, and 51.6 (±18.8) min in group II, showing a statistically significant difference (p

Single nucleotide polymorphisms (SNPs) are the most abundant forms of human genetic variations and resources for mapping complex genetic traits and disease association studies. We have constructed a linkage disequilibrium (LD) map of chromosome 22 in Korean samples and compared it with those of other populations, including Yorubans in Ibadan, Nigeria (YRI), Centre d'Etude du Polymorphisme Humain (CEPH) reference families (CEU), Japanese in Tokyo (JPT) and Han Chinese in Beijing (CHB) in the HapMap database. We genotyped 4681 of 111,448 publicly available SNPs in 90 unrelated Koreans. Among genotyped SNPs, 4167 were polymorphic. Three hundred and five LD blocks were constructed to make up 18.6% (6.4 of 34.5 Mb) of chromosome 22 with 757 tagSNPs and 815 haplotypes (frequency ≥ 5.0%). Of 3430 common SNPs genotyped in all five populations, 514 were monomorphic in Koreans. The CHB + JPT samples have more than a 72% overlap with the monomorphic SNPs in Koreans, while the CEU + YRI samples have less than a 38% overlap. The patterns of hot spots an d LD blocks were dispersed throughout chromosome 22, with some common blocks among populations, highly concordant between the three Asian samples. Analysis of the distribution of chimpanzee-derived allele frequency (DAF), a measure of genetic differentiation, Fst levels, and allele frequency difference (AFD) among Koreans and the HapMap samples showed a strong correlation between the Asians, while the CEU and YRI samples showed a very weak correlation with Korean samples. Relative distance as a quantitative measurement based upon DAF, Fst, and AFD indicated that all three Asian samples are very proximate, while CEU and YRI are significantly remote from the Asian samples. Comparative genome-wide LD studies provide useful information on the association studies of complex diseases.

OBJECTIVES :A2 –3-cm blowhole incision in the supraclavicular or infraclavicular area is widely used to eliminate the presence of subcutaneous air in cases of life-threatening subcutaneous emphysema (SE). However, when the patient is supported by mechanical ventilation, it is difficult to eliminate completely such air because mechanical ventilation leads consistently to the formation of large amounts of air. To overcome this, we applied negative pressure wound therapy (NPWT) along with blowhole incisions for the treatment of severe SE. METHODS: To evaluate the feasibility of NPWT, we retrospectively analysed the clinical outcomes of 10 patients who developed severe SE during ventilator care and were treated with a modified blowhole incision using NPWT from January 2009 to November 2013. RESULTS: All patients showed immediate improvement in SE after NPWT, and no symptom aggravation occurred after NPWT. The mean duration of NPWT was 7.5 ± 5.1 (range, 3–14) days, and the mean number of dressing changes was 1.5 ± 0.7 (range, 1–2). There were no blowhole-incision-related wound infections or any other complications. CONCLUSIONS: While it is not necessary to apply a blowhole incision with NPWT in all cases of SE, this therapy can be helpful for patients with severe SE associated with mechanical ventilation requiring rapid decompression.

The leukotrienes (LTs) enhance allergen- and interleukin (IL)-13-dependent allergic lung inflammatory disease. However, the precise requirement of LTs and the mechanism by which they elicit allergic lung responses remain uncertain. To clarify the involvement of LTs in respiratory allergen- and IL-13-induced experimental asthma and elucidate the underlying mechanisms of LTs-mediated enhanced allergic asthma, we investigated the role of LTs in two models of allergic inflammation: intranasal Aspergillus protease allergen and recombinant IL-13-induced T helper type 2 (Th2) cell-mediated inflammation, and also examined Th2-related chemokines downstream of LTs signaling. 5-Lipoxygenase (5-LO)-deficient mice exposed to short-term intranasal Aspergillus protease allergen showed attenuated airway inflammation, decreased airway hyper-responsiveness and reduced bronchoalveolar eosinophilia when compared to wild-type mice. However, this phenotype was less apparent using long exposure to the same allergen. 5-LO-deficient mice exposed to intranasal rIL-13 also showed attenuated phenotypes of allergic asthma via significant reduction in Th2-specific chemokines, CCL7 and CCL17 production and decreased Th2 cells recruitment to the lungs. Addition of leukotriene B4 (LTB4) and LTC4 to the airways of 5-LO-deficient mice resulted in the rescue of rIL-13-induced experimental asthma. Furthermore, LTs addition to rIL-13 synergistically enhanced the production of Th2-specific chemokines in the lung and inflammatory responses. Therefore, our findings suggest that LTs complement allergens and their downstream cytokine (e.g., IL-13) induced Th2 inflammation by enhancing the induction of Th2 chemokines.

Eosinophilic pleural effusion (EPE) is characterized by greater than 10% eosinophilia and is frequently associated with air and/or blood in the pleural cavity. Primary spontaneous pneumothorax (PSP), defined as the spontaneous presence of air in the pleural space, is one of the most common causes of EPE. Recent studies have shown that type 2 immune responses play important roles in eosinophilic airway inflammation resulting in pleural pathology.To determine the predominant immune responses associated with PSP in humans, and to examine whether IL-33, thymic stromal lymphopoietin (TSLP), or type 2 innate lymphoid cell (ILC2)-mediated immune responses are associated factors.Eosinophil-associated cytokines were measured in the pleural fluid of patients with PSP and control subjects. Th2 cell and ILC2 responses in the pleural cavity and peripheral blood were also evaluated by in vitro restimulation and intracellular cytokine staining of T cells and ILC2s in patients with PSP (n = 62) and control subjects (n = 33). IL-33-mediated IL-5 production by ILC2s was also evaluated.Significantly higher concentrations of IL-5 and eotaxin-3 were detected in the pleural fluid of patients with PSP, in addition to significantly higher concentrations of IL-33 and TSLP. Although IL-5 production was induced by IL-33 treatment of ILC2s, other Th2 cell-mediated immune responses were not detected.Our results indicate that innate immune responses characterized by the production of IL-33, TSLP, and IL-5 are associated with the development of EPE in PSP by an ILC2-dependent and Th2-independent mechanism.

Increased rat renal ammoniagenesis is sustained during chronic metabolic acidosis by the cell-specific induction of the regulatory enzymes of glutamine catabolism and of gluconeogenesis. A glutaminase-specific cDNA hybridizes to 6.0- and 3.4-kb mRNAs that are contained in total or poly(A)+ RNA isolated from rat kidney. When translated in a rabbit reticulocyte lysate, each of the fractionated mRNAs produces the 72-kDa precursor of the mitochondrial glutaminase. The levels of both mRNAs are increased 5-fold within 1 day following onset of chronic acidosis and reach a maximum (8-fold) after 5 days. During recovery from chronic acidosis, the levels of the glutaminase mRNAs are returned to normal within 1 day. The observed changes in mRNA levels correlate with equivalent changes in the relative levels of translatable glutaminase mRNA. Nuclear run-on assays indicate that the rate of transcription of the renal phosphoenolpyruvate carboxykinase gene is increased and decreased in response to onset and recovery from chronic acidosis, respectively. In contrast, the rates of transcription of the glutaminase and beta-actin genes are unaffected by alterations in acid-base balance. Thus, the increase in renal glutaminase activity during chronic acidosis results from an equivalent increase in the levels of total and translatable glutaminase mRNAs which apparently results from an increased stability of the glutaminase mRNA.

During chronic acidosis, the levels of the rat renal mRNAs that encode the mitochondrial glutaminase (GA) and cytosolic phosphoenolpyruvate carboxykinase (PCK) are increased 6-fold. Following acute recovery of chronic acidosis, the levels of the two mRNAs are rapidly and coordinately decreased, returning to normal within 13-17 h. In contrast, the increases in GA and PCK mRNAs during acute onset of acidosis occur with very different kinetics. The increase in PCK mRNA occurs rapidly and reaches a maximum within 7 h, whereas the GA mRNA is increased after a 4-7-h lag and then plateaus at 14-17 h. Treatment with dexamethasone or with cAMP analogs significantly increases the level of renal PCK mRNA but has no effect on the level of GA mRNA. Nuclear run-on experiments indicate that the acute induction of PCK mRNA is primarily due to an increased rate of transcription. However, transcription of GA mRNA is unaffected by acute acidosis. Therefore, the changes in the two mRNAs are temporally coordinated but occur through different mechanisms. Furthermore, the inductive effects of acidosis are not mediated solely through glucocorticoid or cAMP regulatory elements.

Introduction: Anatomical variation of the sympathetic nervous system is known to be one of the main causes of failure and dissatisfaction after sympathetic surgery. However, there are onlyfew reportson the descriptive analysis of sympathetic nerve variants. The purposeof this studyis to investigate the anatomical variations of the sympathetic trunk at the levels of T3 and T4 ganglia considered in a topographic approach for sympatheticproceduresandtofurtherimprovethepostoperativeoutcome.Materialsandmethods:FromJune2003toJanuary2004,44patients with palmar hyperhidrosis underwent bilateral T3,4 ramicotomy via video-assisted thoracoscopic surgery. The anatomy of T3 and T4 sympathetic ganglia, pathway of sympathetic trunk, and rami-communicantes were recorded on video and still cut images for descriptive analysis. Results: The thoracic sympathetic trunks were mostly lying against the heads of the ribs, but there were variants of sympathetic trunk running along the medial side of the rib heads of 3rd, 4th and 5th ribs, respectivelyin 9.0%, 18.0% and 37.5% of the cases. There were also variants running along the lateral side of rib heads near the neck portion in 12.5%, 10.2% and 8.0% of the cases. The 3rd ganglion was located within the intercostal space (59.1%) or at the level of the upper border of the 4th rib (36.4%) or upon the 4th rib (4.5%). The location of the 4th ganglion was in the intercostal space(18.2%),theupperborderofthe5thrib(44.3%)oruponthe5thrib(37.5%).Theascendingramiwerefoundatthelevelofthe3rdganglionin 48.8% and the 4th ganglion in 45.5% of the cases. The descending rami were located at the level of 3rd and the 4th ganglion in 8.0% and 6.8%, respectively. And the middle rami were found in all cases except one. Conclusions: It may be difficult to localize the sympathetic trunk in some casesofsevereobesity; acarefulinspectionhastobeperformedfromthemedialsideoftheribheadstotheneckportion.Theobvious‘downward shift of ganglion’ in the position shown as the thoracic sympathetic trunk descends is to be deliberated in T4 sympathetic surgery. Many ascending and descending accessory pathways of sympathetic nerve were observed; therefore, a lateral extension of electrocoagulation at the level of upper and lower rib border is necessary to impose a complete blockage of sympathetic nerve stimulus. # 2008 European Association for Cardio-Thoracic Surgery. Published by Elsevier B.V. All rights reserved.

Lung transplantation is a viable option for patients with chronic obstructive pulmonary disease (COPD), and emphysema is the most common indication to undergo lung transplantation. A total of seven lung and one heart-lung transplantations were performed between July 1996 and June 2004 at the Yongdong Severance Hospital, and herein, three emphysema patients who underwent single lung transplantations are reviewed. There were 2 males and 1 female, with a mean age of 50 years (35, 57 and 58 years). They all underwent an operation, without cardiopulmonary bypass, and there was no operative mortality. The mean survival was 12 months (4 months, 15 months and 17 months) and all succumbed to death due to activation of pulmonary tuberculosis, post-transplantation lymphoproliferative disease and cytomegalovirus (CMV) gastritis associated with asphyxia. Infection was the most common postoperative complication, resulting in longer hospital stays, higher medical expenses and shorter survival rates, necessitating aggressive prophylactic management. The accumulation of experience, modifications to operative procedures and perioperative care may lead to improved early and long-term survival in patients with emphysema undergoing single or bilateral lung transplantations.

The Nuss procedure is a newly developed operative method for minimally invasive repair of pectus excavatum in pediatric patients. However, the surgical indication for this procedure has been extended into adult patients. The aim of this study was to assess the surgical outcome of the Nuss procedure in different age groups and to analyze its feasibility in the adult population.From December 1999 to March 2003, 51 patients (40 males and 11 females) with pectus excavatum underwent the Nuss procedure. We classified patients into three groups based on age (pediatric, adolescent, and adult), retrospectively analyzed demographic, intraoperative and postoperative data, and compared outcomes among each group.Mean operation time was 52.0 +/- 22.9 minutes, 80.4 +/- 27.4 minutes, and 127.3 +/- 44.9 minutes in the pediatric, adolescent, and adult groups, respectively (p0.001). Postoperative complications occurred in 3 of 27 patients (11.1%) in the pediatric group and in 7 of 12 patients (58.3%) in both the adolescent and adult groups (p = 0.002). Reoperations were performed due to complications in 1 of 27 patients (3.7%) in the pediatric group, 2 of 12 patients (16.6%) in the adolescent group, and 5 of 12 patients (41.7%) in the adult group (p = 0.001).The Nuss procedure is highly recommended in pediatric patients with pectus excavatum. However, in adults it is necessary to select patients carefully because of the longer operation time and higher incidence of complications associated with the procedure in this population.

One of the major challenges in genome research is the identification of the complete set of genes in a genome. Alignments of expressed sequences (RNA and EST) with genomic sequences have been used to characterize genes. However, the number of alignments far exceeds the likely number of genes in a genome, suggesting that, for many genes, two or more alignments can be joined through overlapping sequences to yield accurate gene structures. High-throughput EST sequencing becomes less efficient in closing those alignment gaps due to its nonselective nature. We sought to bridge these alignments through a novel approach: targeted cDNA sequencing. Human expressed sequences from GenBank version 124 were aligned with the genomic sequence from NCBI build 24 using LEADS, Compugen's EST and RNA clustering and assembly software system. Nine hundred forty-eight pairs of alignments were selected based on EST clone information and/or their homology to the same known proteins. Reverse transcriptase PCR and sequencing yielded sequences for 363 of those pairs. These sequences helped characterize over 60 novel or otherwise incomplete genes in the recent UniGene build 153, which included over 1 million additional ESTs. These results indicate that this integrated and targeted strategy, combining computational prediction and experimental cDNA sequencing, can efficiently generate the overlapping sequences and enable the full characterization of genomes. Additional information about the contig pairs, the resultant overlapping sequences, tissue sources, and tissue profiles are available in a supplemental file. The majority of sequences were deposited with GenBank and their accession numbers are within the range BU101504 to BU102193 .

Corneal endothelial cells have a limited capacity for proliferation. Upon transformation with the SV40 large T antigen, however, these cells undergo division and grow rapidly. In order to gain insight into the control mechanisms that determine this proliferative switch, we investigated the expression level and activity of various known cell cycle-regulatory proteins in these cells. Primary human and rabbit corneal endothelial cells were transduced in vitro with a replication-defective adenovirus containing SV40 large T antigen, and subsequently the expression and activity of cell cycle-regulatory proteins was analyzed. Cells transduced with large T antigen exhibited strongly increased activity of cyclin-dependent kinases. This increase correlated with the elevated expression of various cyclin-dependent kinase subunits, such as cyclin A, and to a lesser extent, cyclin D, cdk2, and cdk4. Furthermore, the expression of two cyclin-dependent kinase inhibitors, p21(WAF1) and p27(KIP1), which was high in primary human cells (but not in primary rabbit cells), was strongly reduced in large T-antigen transduced cells. Thus, the remarkably low proliferative activity of normal human corneal endothelial cells appears to be regulated at two levels: the expression of certain cell cycle-regulatory proteins that are essential for cell cycle progression is extremely low (cyclin A) or somewhat low (cdk2 and cdk4); but the amount of p21 and p27, inhibitors of cell cycle progression, is very high. As a consequence, the enzymatic activity of cyclin-dependent kinase is below detectable levels. However, the growth-inhibitory status of these components is clearly reversible: upon transduction with large T antigen, the expression of cyclin A, cyclin D, cdk2, and cdk4 is induced, whereas the expression of p21 and p27 is inhibited, and the cells proliferate. Thus, our study provides insight into the molecular basis of the attenuated proliferation of corneal endothelial cells and suggests potential targets that could be manipulated for the purpose of therapeutic interventions aimed at renewed cell growth.

The human MAT1 gene (menage a trois 1) is an assembly factor and a targeting subunit of cyclin-dependent kinase (CDK)-activating kinase. The novel mechanisms by which MAT1 forms an active CDK-activating kinase and determines substrate specificity of CDK7-cyclin H are involved in the cell cycle, DNA repair, and transcription. Hyperplasia of vascular smooth muscle cells (SMC) is a fundamental pathologic feature of luminal narrowing in vascular occlusive diseases, and nothing is yet known regarding the cell cycle phase specificity of the MAT1 gene in its involvement in SMC proliferation. To investigate such novel regulatory pathways, MAT1 expression was abrogated by retrovirus-mediated gene transfer of antisense MAT1 RNA in cultured rat aortic SMCs. We show that abrogation of MAT1 expression retards SMC proliferation and inhibits cell activation from a nonproliferative state. Furthermore, we have demonstrated that these effects are due to G1 phase arrest and apoptotic cell death. Our studies indicate a link between cell cycle control and apoptosis and reveal a potential mechanism for coupling the regulation of MAT1 with G1 exit and prevention of apoptosis.

Thoracoscopic plication is the most effective treatment for diaphragmatic enventration. However, the conventional thoracoscopic plication procedure introduced by Mouroux in 1996 has some disadvantages. We improved and simplified the Mouroux technique with the patient in the head up position, CO(2) insufflations and figure-of-eight sutures. These were possible to perform by pure thoracoscopic surgery using three 5 mm ports without the requirement for open thoracotomy.

Background The prognosis of acute lung injury (ALI) after pneumonectomy is poor, with reported mortality rates of 30-100%. Neutrophil elastase inhibitor (NEI) is known to prevent lung injury caused by neutrophil elastase and improve lung function in ALI. We evaluated the effect of NEI on ALI after pneumonectomy. Methods We analyzed nine patients who required ventilator care due to ALI after pneumonectomy. Five of these patients underwent conventional ventilator care (group I), and four patients underwent ventilator care and were administrated NEI (group II). We retrospectively analyzed the lung injury score (LIS) for 10 days after intubation. Results The LIS before intubation satisfied the diagnostic criteria of ALI or acute respiratory distress syndrome (ARDS) in all patients. After intubation, the LIS improved in both groups. But, as times went on, the mean value of the LIS in group II was lower compared to group I. In group I, only one patient underwent extubation. In group II, extubation was possible in three patients. Mortality rates were 80% in group I and 25% in group II. Conclusions We conclude that NEI may improve the lung function, shorten the duration of mechanical ventilation, and reduce mortality in patients with ALI after pneumonectomy.

We describe the molecular cloning of two novel human and murine transcription factors containing the TEA/ATTS DNA binding domain and related to transcriptional enhancer factor-1 (TEF-1). These factors bind to the consensus TEA/ATTS cognate binding site exemplified by the GT-IIC and Sph enhansons of the SV40 enhancer but differ in their ability to bind cooperatively to tandemly repeated sites. The human TEFs are differentially expressed in cultured cell lines and the mouse (m)TEFs are differentially expressed in embryonic and extra-embryonic tissues in early post-implantation embryos. Strikingly, at later stages of embryogenesis, mTEF-3 is specifically expressed in skeletal muscle precursors, whereas mTEF-1 is expressed not only in developing skeletal muscle but also in the myocardium. Together with previous data, these results point to important, partially redundant, roles for these TEF proteins in myogenesis and cardiogenesis. In addition, mTEF-1 is strongly coexpressed with mTEF-4 in mitotic neuroblasts, while accentuated mTEF-4 expression is also observed in the gut and the nephrogenic region of the kidney. These observations suggest additional roles for the TEF proteins in central nervous system development and organogenesis.

A single phage was isolated from a λ gt11 rat brain cDNA library by screening with antibodies prepared against rat renal glutaminase. Partial proteolysis of the fusion protein produced by a lysogen of the isolated phage generated a series of immunoreactive peptides that co-migrated with those derived from the purified brain glutaminase. The cDNA has a single open reading frame which encodes 326 amino acids that are in frame with β-galactosidase. A 72-kDa protein, corresponding in size to the precursor of mitochondrial glutaminase, was immunoprecipitated from the translation products of rat renal mRNA that selectively hybridized to the cDNA. A probe made from the glutaminase cDNA detected an mRNA about 6 kb in length. This mRNA was present in rat brain and normal kidney RNA, increased 6-fold in acidotic kidney RNA, but was not detectable in liver RNA.

The regions of transcriptional enhancer factor-1 (TEF-1) required for its activation function and sequence-specific DNA binding have been determined. Deletion analysis of a chimera between TEF-1 and the GAL4 DNA binding domain (DBD) indicated that at least three regions of TEF-1 were involved in transactivation. However, none of these regions functioned as independent activating domains. Moreover, none of the GAL4 chimeras containing individual TEF-1 regions interfered with the activity of endogenous HeLa cell TEF-1, while interference was observed with the GAL4-TEF-1 chimeras which functioned as transactivators. These results indicate that there is a general correlation between the abilities of a given GAL4-TEF-1 chimera to function in transcriptional activation and interference, thus supporting the idea that transactivation by TEF-1 is mediated by a limiting transcriptional intermediary factor. In addition, we show experimentally that the TEA/ATTS domain is a novel class of DBD involved in the sequence-specific DNA binding of TEF-1 and its Drosophila homologue scalloped. Two other regions of TEF-1 are also required for DNA binding. These regions are not part of the minimum DBD, but may function by antagonizing the effect of sequences which negatively regulate DNA binding mediated by both the TEF-1 TEA/ATTS domain and the GAL4 DBD. In addition, analysis of TEF-1 and scalloped derivatives in which their TEA/ATTS domains have been interchanged further indicates that the TEA/ATTS domain is not the only determinant of DNA binding specificity.