Treatment of limbic encephalitis with anti-glioma-inactivated 1 (LGI1) antibodies.

We report a 72-year-old patient who developed acute limbic encephalitis initially considered of uncertain aetiology. Detailed information on clinical presentation, MRI appearance, antibody levels, cognitive impairment assessment, treatment and evolution of the patient is reported here. Since the early 2000s, many antibodies implied in central nervous system autoimmune disorders have been identified. Anti-glioma-inactivated 1 (LGI1) antibodies have been recently identified as associated with limbic encephalitis, as was the case in our patient.

A human RNA polymerase II subunit is encoded by a recently generated multigene family.

BACKGROUND: The sequences encoding the yeast RNA polymerase II (RPB) subunits are single copy genes. RESULTS: While those characterized so far for the human (h) RPB are also unique, we show that hRPB subunit 11 (hRPB11) is encoded by a multigene family, mapping on chromosome 7 at loci p12, q11.23 and q22. We focused on two members of this family, hRPB11a and hRPB11b: the first encodes subunit hRPB11a, which represents the major RPB11 component of the mammalian RPB complex; the second generates polypeptides hRPB11balpha and hRPB11bbeta through differential splicing of its transcript and shares homologies with components of the hPMS2L multigene family related to genes involved in mismatch-repair functions (MMR). Both hRPB11a and b genes are transcribed in all human tissues tested. Using an inter-species complementation assay, we show that only hRPB11balpha is functional in yeast. In marked contrast, we found that the unique murine homolog of RPB11 gene maps on chromosome 5 (band G), and encodes a single polypeptide which is identical to subunit hRPB11a. CONCLUSIONS: The type hRPB11b gene appears to result from recent genomic recombination events in the evolution of primates, involving sequence elements related to the MMR apparatus.

Isolation and characterization of monoclonal antibodies directed against subunits of human RNA polymerases I, II, and III.

Human nuclei contain three different RNA polymerases: polymerases I, II, and III. Each polymerase is a multi-subunit enzyme with 12-17 subunits. The localization of these subunits is limited by the paucity of antibodies suitable for immunofluorescence. We now describe eight different monoclonal antibodies that react specifically with RPB6 (also known as RPA20, RPB14.4, or RPC20), RPB8 (RPA18, RPB17, or RPC18), RPC32, or RPC39 and which are suitable for such studies. Each antibody detects one specific band in immunoblots of nuclear extracts; each also immunoprecipitates large complexes containing many other subunits. When used for immunofluorescence, antibodies against the subunits shared by all three polymerases (i.e., RPB6, RPB8) gave a few bright foci in nucleoli and nucleoplasm, as well as many fainter nucleoplasmic foci; all the bright foci were generally distinct from speckles containing Sm antigen. Antibodies against the two subunits found only in polymerase III (i.e., RPC32, RPC39) gave a few bright and many faint nucleoplasmic foci, but no nucleolar foci. Growth in two transcriptional inhibitors-5, 6-dichloro-1-beta-d-ribofuranosylbenzimidazole and actinomycin D-led to the redistribution of each subunit in a characteristic manner.

EWS, but not EWS-FLI-1, is associated with both TFIID and RNA polymerase II: interactions between two members of the TET family, EWS and hTAFII68, and subunits of TFIID and RNA polymerase II complexes.

The t(11;22) chromosomal translocation specifically linked to Ewing sarcoma and primitive neuroectodermal tumor results in a chimeric molecule fusing the amino-terminus-encoding region of the EWS gene to the carboxyl-terminal DNA-binding domain encoded by the FLI-1 gene. As the function of the protein encoded by the EWS gene remains unknown, we investigated the putative role of EWS in RNA polymerase II (Pol II) transcription by comparing its activity with that of its structural homolog, hTAFII68. We demonstrate that a portion of EWS is able to associate with the basal transcription factor TFIID, which is composed of the TATA-binding protein (TBP) and TBP-associated factors (TAFIIs). In vitro binding studies revealed that both EWS and hTAFII68 interact with the same TFIID subunits, suggesting that the presence of EWS and that of hTAFII68 in the same TFIID complex may be mutually exclusive. Moreover, EWS is not exclusively associated with TFIID but, similarly to hTAFII68, is also associated with the Pol II complex. The subunits of Pol II that interact with EWS and hTAFII68 have been identified, confirming the association with the polymerase. In contrast to EWS, the tumorigenic EWS-FLI-1 fusion protein is not associated with either TFIID or Pol II in Ewing cell nuclear extracts. These observations suggest that EWS and EWS-FLI-1 may play different roles in Pol II transcription.

Interactions between the human RNA polymerase II subunits.

As an initial approach to characterizing the molecular structure of the human RNA polymerase II (hRPB), we systematically investigated the protein-protein contacts that the subunits of this enzyme may establish with each other. To this end, we applied a glutathione S-transferase-pulldown assay to extracts from Sf9 insect cells, which were coinfected with all possible combinations of recombinant baculoviruses expressing hRPB subunits, either as untagged polypeptides or as glutathione S-transferase fusion proteins. This is the first comprehensive study of interactions between eukaryotic RNA polymerase subunits; among the 116 combinations of hRPB subunits tested, 56 showed significant to strong interactions, whereas 60 were negative. Within the intricate network of interactions, subunits hRPB3 and hRPB5 play a central role in polymerase organization. These subunits, which are able to homodimerize and to interact, may constitute the nucleation center for polymerase assembly, by providing a large interface to most of the other subunits.

The gene (POLR2L) encoding the hRPB7.6 subunit of human RNA polymerase.

The gene (POLR2L) encoding a 7.6-kDa subunit (hRPB7.6) of human RNA polymerase has been cloned. It compromises two exons, 116 and 227 bp, respectively, interspaced with an intron of about 2.1 kb. This gene, whose localization has been assigned to the short arm of chromosome 11 (position 11p15), is transcribed in HeLa cells as one major messenger RNA, which encodes a 67-residue polypeptide (7645 Da) that shares strong homologies with the corresponding subunits of other eukaryotic and archaeal RNA polymerase subunits. Like its yeast counterpart (ABC10 beta, encoded by the RPB10 gene), the hRPB7.6 subunit may be shared by all three classes of human nuclear RNA polymerase. Cysteine residues characteristic of an atypical zinc-binding domain are conserved in the homologous sequences of all six species analyzed. A small, related RNA polymerase subunit from vaccinia virus exhibits an identical set of cysteines, suggesting that these residues may be contribute to a crucial function in the multimeric RNA polymerases.

Four subunits that are shared by the three classes of RNA polymerase are functionally interchangeable between Homo sapiens and Saccharomyces cerevisiae.

Four cDNAs encoding human polypeptides hRPB7.0, hRPB7.6, hRPB17, and hRPB14.4 (referred to as Hs10 alpha, Hs10 beta, Hs8, and Hs6, respectively), homologous to the ABC10 alpha, ABC10 beta, ABC14.5, and ABC23 RNA polymerase subunits (referred to as Sc10 alpha, Sc10 beta, Sc8, and Sc6, respectively) of Saccharomyces cerevisiae, were cloned and characterized for their ability to complement defective yeast mutants. Hs10 alpha and the corresponding Sp10 alpha of Schizosaccharomyces pombe can complement an S. cerevisiae mutant (rpc10-delta::HIS3) defective in Sc10 alpha. The peptide sequences are highly conserved in their carboxy-terminal halves, with an invariant motif CX2CX12RCX2CGXR corresponding to a canonical zinc-binding domain. Hs10 beta, Sc10 beta, and the N subunit of archaeal RNA polymerase are homologous. An invariant CX2CGXnCCR motif presumably forms an atypical zinc-binding domain. Hs10 beta, but not the archaeal subunit, complemented an S. cerevisiae mutant (rpb10-delta 1::HIS3) lacking Sc10 beta. Hs8 complemented a yeast mutant (rpb8-delta 1::LYS2) defective in the corresponding Sc8 subunit, although with a strong thermosensitive phenotype. Interspecific complementation also occurred with Hs6 and with the corresponding Dm6 cDNA of Drosophila melanogaster. Hs6 cDNA and the Sp6 cDNA of S. pombe are dosage-dependent suppressors of rpo21-4, a mutation generating a slowly growing yeast defective in the largest subunit of RNA polymerase II. Finally, a doubly chimeric S. cerevisiae strain bearing the Sp6 cDNA and the human Hs10 beta cDNA was also viable. No interspecific complementation was observed for the human hRPB25 (Hs5) homolog of the yeast ABC27 (Sc5) subunit.

Chromosomal localization of human RNA polymerase II subunit genes.

The eukaryotic DNA-dependent RNA polymerase II (or B) is composed of 10 to 14 polypeptides ranging from 220 to 10 kDa. To gain further insight into the molecular structure and function of these subunits, we have undertaken the molecular cloning of nucleotide sequences corresponding to the human enzyme. The cDNAs of five subunits (hRPB220, hRPB140, hRPB33, hRPB25, and hRPB14.5) have been isolated. Using in situ hybridization, we show that the genes of these subunits have distinct chromosomal locations (17p13, 4q12, 16q13-q21, 19p13.3, and 19q12, respectively). Thus, if assembly of active polymerase molecules requires coordinated expression from these independent genes, mechanisms that ensure tight coregulation of the corresponding promoters must exist.

Structure of the gene encoding the 14.5 kDa subunit of human RNA polymerase II.

The structure of the gene encoding the 14.5 kDa subunit of the human RNA polymerase II (or B) has been elucidated. The gene consists of six exons, ranging from 52 to over 101 bp, interspaced with five introns ranging from 84 to 246 bp. It is transcribed into three major RNA species, present at low abundance in exponentially growing HeLa cells. The corresponding messenger RNAs contain the same open reading frame encoding a 125 amino acid residue protein, with a calculated molecular weight of 14,523 Da. This protein (named hRPB14.5) shares strong homologies with the homologous polymerase subunits encoded by the Drosophila (RpII15) and yeast (RPB9) genes. Cysteines characteristic of two zinc fingers are conserved in all three corresponding sequences and, like the yeast protein, the hRPB14.5 subunit exhibits zinc-binding activity.

Primary structure of the second largest subunit of human RNA polymerase II (or B).

The cDNA of the second largest subunit of RNA polymerase II (or B) from HeLa cells has been cloned and sequenced. A predicted amino acid sequence of 1174 residues (calculated molecular mass of 133,896 Da) was derived from the longest open reading frame and compared to the sequences of homologous subunits of polymerases of eukaryotic, archaeal and bacterial origin. After optimal alignment, about 16% of the residues were found to be conserved throughout evolution, from human to Escherichia coli. About 2/3 of the overall length of the conserved domains delineated by these residues are clustered within the C-terminal half of the human polypeptide, whereas the remaining is spread over its N-terminal half. The putative functional significance of these conserved domains is discussed.

[Causes of failure analysis after cardiac transplantation. From a consecutive series of 91 cases]. / Analyse des causes d'échec après transplantation cardiaque. D'après une série consécutive de 91 cas.

Ninety-three cardiac transplantations were carried out in 91 patients (2 retransplantations) between March 1st 1987 and November 1st 1989, in 84 adults and 7 children under 15 years of age. The indications were dilated cardiomyopathy (48%), ischemic cardiomyopathy (35%), decompensated valvular heart disease (11%), congenital heart disease (3%) and two cases of Uhl's anomaly. Twelve patients underwent transplantation after external circulatory assistance (13%), 11 patients after inscription on the list of extreme emergencies, and 68 on an elective basis (74%). The postoperative immunosuppressive protocol was triple therapy: Ciclosporine, Azathioprine and Prednisone. Three of the children died. The early adult mortality was 9 cases (10.7%). It was 8% in patients operated electively. Major infectious complications occurred in 10 patients (11%). Rejection was looked for by systematic endomyocardial biopsy and echocardiography. Three hundred and forty-nine biopsies were made. Thirty-five patients (44%) had no problems of rejection. Seventy-nine patients have now been followed up for an average of 19 months. There were 7 late deaths. Seventy seven per cent of the survivors are asymptomatic. Acute rejection and transplant dysfunction were the two main causes of early mortality after cardiac transplantation. Although the long-term prognosis is uncertain, the medium-term results are very encouraging.

[Saint-Jude Medical tricuspid prosthesis. Long-term clinical, biological and echocardiographic assessment]. / La prothèse Saint-Jude Médical en position tricuspidienne. Evaluations clinique, biologique et échographique à long terme.

Twenty-four patients with a Saint-Jude Medical tricuspid valve prosthesis, aged 5 to 77 years, were studied. The etiology of the tricuspid lesion was rhumatic in 17 cases, infectious in 4 cases, and congenital in the other 3. Fourteen patient (58%) had undergone previous valve surgery, 7 of whom had undergone tricuspid valve replacement (TVR) by a bioprosthesis. Three patients were operated on for the third time. The TVR was isolated (4 cases) or associated with aortic valve replacement (3 cases), mitral valve replacement (8 cases), double aortic and mitral valve replacement (7 cases), repair of a ventricular septal defect (VSD) (1 case) and radical treatment of a Wolff-Parkinson-White syndrome (WPW) in 1 case. There were 3 early deaths (12.5%). Eighteen of the 21 survivors were followed up clinically, biologically (detection of hemolysis) and by Doppler echocardiography for an average period of 45 months (range 10 to 96 months). The clinical benefit was clear cut. No embolic complications were observed and there were no cases of hemolysis. The mean resting tricuspid pressure gradient was 3.57 +/- 2.36 mmHg. The Saint-Jude Medical prosthesis would therefore seem to be a good alternative to other mechanical valve prosthesis in the tricuspid position and without the risk of valve degeneration associated with bioprosthesis.

[Treatment using traditional acupuncture of early scapulohumeral pains following heart surgery]. / Traitement par acupuncture traditionnelle des algies scapulo-humérales précoces après chirurgie cardiaque.

The purpose of this study was to test the efficacy of traditional Chinese acupuncture in the treatment of scapulohumeral pain during the early stage following heart surgery, by puncture of points not related anatomically or metamerically with the scapulohumeral joint and without any needle stimulation. Reduction of pain and angular gain were almost immediate, durable, measurable and reproducible, which could be explained by possible effects of acupuncture on articular sympathetic mechanoreceptors, then suppressing reflex muscular contractions due to intraoperative postural constraints.

[Abnormal origin of the left coronary artery in the adult. Scintigraphic and surgical correlations]. / Naissance anormale de la coronaire gauche chez l'adulte. Corrélations scintigraphique et chirurgicale.

The authors report two cases of anomalous origin of the left coronary artery from the pulmonary artery in the adult. The two patients were pauci-symptomatic and were successfully operated, the one by reimplantation of the left coronary artery in the aorta and the other by an internal mammary artery left anterior descending artery bypass. Resting and stress myocardial scintigraphy and radionuclide ventriculography were performed before and after surgery in both cases. An analysis of segmental wall motion was possible in one patient. Before surgery, there was hypo-fixation of the tracer during the stress test and an alteration of left ventricular function. Postoperative isotopic investigations confirmed the efficacy of surgery the absence of regional ischemia and the normalisation of the ventricular contraction. These results argue in favour of a surgical reconstruction of a two coronary system, given the spontaneous risk of sudden death in this condition.

Use of the Abiomed BVS System 5000 as a bridge to cardiac transplantation.

The Abiomed BVS System 5000 (Abiomed Cardiovascular, Inc., Danvers, Mass.) is a gravity-filled, pneumatically driven external prosthetic ventricle that has been implanted as a circulatory support device in six patients 9 to 58 years of age, presenting with a refractory heart failure nonamenable to any type of corrective operation. Three (including a 9-year-old girl) had an end-stage nonobstructive myocardiopathy, and two (including one patient who had had a massive recent myocardial infarction) had an ischemic heart disease. When first seen, the 58-year-old patient had an acute rejection and graft failure occurring 2 months after a first transplantation. All patients showed evidence of a low-output state (cardiac index less than 1.5 L/min/m2), with renal failure (mean urinary output, less than 27 ml/min) and hypoxia (mean arterial oxygen pressure = 56 torr under 80% forced inspiratory oxygen), despite maximum pharmacologic support (dobutamine, 16 to 18 gamma/kg/min; dopamine, 3 to 18 gamma/kg/min; adrenaline, 0.2 to 0.7 gamma/kg/min; furosemide, 7 to 17 gamma/kg/min). The device was implanted through a midline sternotomy and under peripheral normothermic bypass. Five patients received a biventricular support, and one a single left prosthetic ventricle. The cannulation included a right-angled cannula in both the left and right atrium and a suture of the arterial Dacron tubes onto the ascending aorta and main pulmonary artery. After careful deairing of the tubing and ventricles, the console was activated and the bypass progressively discontinued. Heparin infusion was begun 3 hours after chest closure and was continued for the duration of assist pumping, which was 2 to 11 days (mean duration, 7.43 days). The system could provide a complete support of the circulation with both right and left ventricular index remaining stable at 2.4 to 3 L/min/m2. After a dramatic improvement at the time of the system activation, the urinary output remained adequate, thus allowing for a decreasing need for diuretic therapy. In two cases, including one of isolated left ventricular assist pumping, the circulation could be totally supported during 11 hours and 23 hours, respectively, of refractory ventricular tachycardia. Four of six patients were shortly weaned from inotropic agents. Hematologic studies showed a moderate decrease of the coagulation factors level during the first 6 hours of circulatory support, and this remained stable and within normal limits thereafter. There have been three cases of bleeding complications necessitating surgical revision on the sixth hour, the twelfth hour, and the sixth day, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

[Treatment of ventriculo-pulmonary disconnections with prosthetic conduits. Long-term results]. / Traitement des discontinuités ventriculo-pulmonaires par utilisation d'un tube valvulé. Résultats à long terme.

Between 1973 and 1989, 81 consecutive patients aged 2 to 42 years old, with ventriculo-pulmonary discontinuity, were treated by implantation of prosthetic conduits. The initial pathology was Tetralogy of Fallot (33%), complete transposition of the great arteries (20%), truncus arteriosus (17%), double outlet right ventricle (17%) and atrioventricular discordance with L malposition of the great arteries (10%). The overall early mortality was 22% (18 cases) and 14% (5 cases) in the 36 patients operated after 1982. Sixty three patients were followed up for 3 months to 16 years; there were 8 late deaths which occurred spontaneously or at reoperation. Postoperative catheterisation was carried out in 33 cases; the average ventriculopulmonary systolic pressure gradient was 40 +/- 26 mmHg. Six patients were reoperated to change the conduit, on average 6 years +/- 23 months after the first operation. Five other patients underwent endoluminal dilatation of a stenosed conduit which delayed reoperation to change the conduit in 3 cases. Prosthetic conduits have been extensively used in patients with ventriculo-pulmonary discontinuity because they are readily available. However, because of progressive degradation of the prostheses between the 5th and 10th postoperative years, other therapeutic solutions should be considered, i.e. endoventricular repair when possible and, in other cases, the use of aortic homografts.

[Apico-aortic shunts. Indications, results]. / Les dérivations apico-aortiques. Indications--résultats.

Seven children or adolescents aged 8 to 20 years (average 13.5 years) underwent implantation of an apico-aortic conduit from 1980 to June 1989. Eleven previous operations under cardiopulmonary bypass had been performed. The indications were recurrence of muscular subaortic stenosis and/or congenital valvular stenosis in 3 cases, and stenotic, previously implanted aortic valve prosthesis of small calibre in 4 cases. The preoperative left ventricular-aortic systolic pressure gradient was between 70 and 130 mmHg. The first two patients had a bioprosthetic valvulation and the 5 succeeding patients a St Jude Medical mechanical prosthesis. The early and late mortality was nil. The average follow-up period is 5, 6 years at present (range 6 months to 9 years). One patient had to undergo repeat valve replacement after 5 years for degenerescence of the porcine bioprosthesis implanted in the conduit. No other complications related to the conduit or valve were observed. At the endpoint of the study all patients were asymptomatic without treatment. Control echocardiographic data showed normalisation of the indices of left ventricular function. Apico-aortic conduits would seem to be a safe and effective technique for the treatment of recurrent severe obstruction of the left ventricular outflow tract.