A 6-kb EcoRI genomic DNA fragment of Coxiella burnetii, isolated from a recombinant bacteriophage lambda ZapII library, allowed heterologous genetic complementation of Escherichia coli deleted for its dnaJ gene. The C. burnetii dnaJ gene was expressed in E. coli and identified by Western blot analysis using polyclonal antibodies raised against purified E. coli DnaJ protein. Deletion mapping and genetic complementation demonstrated that C. burnetii dnaJ is present on a 2-kb EcoRI-HindIII genomic DNA fragment, from which the nt sequence of the C. burnetii dnaJ gene was determined.

Amino Acid Sequence↗

PubMed1995

Influence of carbon monoxide (CO) on the early course of acute myocardial infarction.

OBJECTIVE: To clarify the influence of an elevated carboxyhemoglobin (COHb) blood level on the course of acute myocardial infarction and to evaluate the administration of supplemental oxygen on the COHb level and the incidence of complications. DESIGN: Prospective clinical study with randomized, unblinded intervention. SETTING: Coronary Care Unit of a university hospital. PATIENTS: 78 consecutive patients with acute myocardial infarction. Excluded were patients with severe dyspnea, pulmonary edema or any other medical indication for supplemental oxygen therapy. INTERVENTIONS: Randomized therapy with 41/min oxygen in 35 patients. MEASUREMENTS AND RESULTS: COHb was measured at admission and 4 h later. The incidence of serious arrhythmias and the maximal creatine kinase (CK) values were recorded. In patients with initial COHb > or = 5%, there were significantly more arrhythmias and significantly higher maximal CK values than in those with normal COHb admission (89 vs 33%, p < 0.001; and 1897 +/- 1602 u/l vs 960 +/- 1097 u/l, p=0.05). This effect was seen only in patients with Q-wave infarction, not in those with non-Q-wave infarction. Supplemental oxygen had no effect on the incidence of arrhythmias. CONCLUSIONS: We conclude that myocardial infarction patients with acute Q-wave infarction and increased COHb levels at admission suffer a more severe course of the disease. This outcome was not influenced by oxygen therapy. Whether this finding indicates a casual relationship and whether higher oxygen concentrations would favorably alter the course of acute myocardial infarction remains to be determined.

Aged↗

PubMed1995

A patient with a high titer of antinuclear antibody and a functioning adrenal tumour.

Reported is the case of a 60-year-old female who was referred to our rheumatology center because of a high titer of antinuclear antibodies (ANAs). Initially several lupus-like features suggested a diagnosis of undifferentiated connective tissue disease (UCTD). Immunofluorescence of the patient's serum showed a nuclear dot pattern. Counterimmunoelectrophoresis for antibodies to Sm, RNP, SS-A and SS-B were negative. Immunoblotting revealed a band at 53 kD which did not correspond to known autoantigens. Several months later the patient presented to the emergency room with symptoms due to a hypertensive crisis. Check-up finally revealed a cortisone-producing tumour of the right adrenal gland. The adenoma was removed by surgery and the patient did well again.

Adrenal Gland Neoplasms↗

PubMed1995

Recurrent cerebrovascular events in patients with patent foramen ovale, atrial septal aneurysm, or both and cryptogenic stroke or transient ischemic attack. French Study Group on Patent Foramen Ovale and Atrial Septal Aneurysm.

Patent foramen ovale (PFO) and atrial septal aneurysm (ASA) have been identified as potential risk factors for stroke, but information about the risk of recurrent cerebral ischemia is scarce. The aim of this retrospective study was to assess the absolute risk of recurrent cerebrovascular events in 132 patients under 60 years of age with patent foramen ovale, atrial septal aneurysm (diagnosed by transesophageal echocardiography with a contrast study), or both and an otherwise unexplained stroke or transient ischemic attack (TIA). During a mean follow-up of 22.6 +/- 16 months, six patients had a recurrent stroke (n = 2) or a TIA (n = 4). No systemic embolism was observed. The actuarial risk of having a recurrent stroke was 2.3% (95% confidence interval, 0.6% to 8.2%) at 2 years, whereas the risk of having a stroke or a TIA was 6.7% (95% confidence interval, 3.1% to 14.2%) at 2 years. The average annual rates of recurrence were 1.2% and 3.4%, respectively. In patients with both PFO and ASA, the actuarial risk of a first recurrent stroke was 9.0% (95% confidence interval, 2.4% to 28.5%) at 2 years, with an average annual rate of recurrence of 4.4%. As a group, patients with patent foramen ovale, atrial septal aneurysm, or both and an otherwise unexplained stroke or TIA appear to have a low risk of recurrent stroke whatever the prophylactic antithrombotic therapy used. The association of ASA and PFO may be an indicator of a higher risk of recurrent stroke.

Adult↗

PubMed1995

Analysis of a Coxiella burnetti gene product that activates capsule synthesis in Escherichia coli: requirement for the heat shock chaperone DnaK and the two-component regulator RcsC.

A 1.2-kb EcoRI genomic DNA fragment of Coxiella burnetti, when cloned onto a multicopy plasmid, was found to induce capsule synthesis (mucoidy) in Escherichia coli. Nucleotide sequence analysis revealed the presence of an open reading frame that could encode a protein of 270 amino acids. Insertion of a tet cassette into a unique NruI restriction site resulted in the loss of induction of mucoidy. Because of its ability to induce mucoidy, we designated this gene mucZ. Computer search for homologies to mucZ revealed 42% identity to an open reading frame located at 1 min of the E. coli chromosome. Interestingly, the C-terminal amino acid residues of MucZ share significant homology with the J domain of the DnaJ protein and its homologs, suggesting potential interactions between MucZ and components of the DnaK-chaperone machinery. Results presented in this paper suggest that E. coli requires DnaK-chaperone machinery for Lon-RcsA-mediated induction of capsule synthesis, as noticed first by S. Gottesman (personal communication). The induction caused by MucZ is independent of Lon-RcsA and is mediated through the two-component regulators RcsC and RcsB. DnaK and GrpE but not DnaJ are also required for the RcsB-mediated MucZ induction, and we propose that MucZ is a DnaJ-like chaperone protein that might be required for the formation of an active RcsA-RcsB complex and for the RcsC-dependent phosphorylation of RcsB. Discussions are presented that suggest three different roles for alternative forms of the DnaK-chaperone machinery in capsule production.

ATP-Dependent Proteases↗

PubMed1995

[Local treatment of breast carcinoma: when is breast-saving therapy not indicated?].

Conservative treatment of breast cancer is now an accepted oncological procedure. It further allows a good functional and cosmetic result. This treatment needs more skills and time than mastectomy, as it does not only include a surgical excision but also a postoperative local radiotherapy and a sometimes difficult follow-up during many years. The contraindications for conservative breast cancer therapy are numerous and depend on factors as tumor, patient and radiotherapy. As more than 1100 patients have been treated since 1977 in our area at a single radiooncological center by a breast conserving procedure. We have confirmed or defined criteria which lead to a good local tumor control as well as to good functional and cosmetic results.

Breast Neoplasms↗

PubMed1995

A human autoantibody recognizing nuclear matrix-associated nuclear protein localized in dot structures.

A human autoimmune serum is used to characterize a protein which is located within a dot-like structure of the interphase nucleus. The dots are located in the nucleoplasma outside the nucleoli. The dot-like structure could be observed on a variety of human substrates (HEp-2, HepG2, HeLa, Molt-4, WI-38 cell lines, peripheral blood lymphocytes), mouse cell lines and tissues (3T3, L929, spleen sections), rat sections, marsupial PtK2 cells and hamster cell lines. The range of the number of dots and their size differed between cells and cell lines and varied between 1 and 24. The antigen could be identified as a 53 kDa protein with a pI of 8.7 and was named NDP53. Digestion experiments suggested that the protein is not associated with DNA or RNA, but is associated with the nuclear matrix. Immunelectron microscopy using ultra-thin sections revealed filamentous structures with a diameter of 0.1 to 0.6 microns. The antigen colocalizes with Sp100, PML and NDP55, which are part of a multiprotein complex known as PML oncogenic domain (POD), nuclear bodies, Kr bodies or ND10.

Animals↗

PubMed1994

[Transesophageal echocardiography in the diagnosis of endocarditis].

Transesophageal echocardiography is a new diagnostic method for visualization of retrocardial structures and heart valves with high resolution. This retrospective analysis describes the use of and the findings by transesophageal echocardiography in patients suspected to suffer from endocarditis studied in three hospitals. 50 patients were studied; the diagnosis was confirmed in 64%, whereas 36% (controls) had other findings. The leading initial symptoms were fever and valvular incompetence. Laboratory parameters were altered in both groups with equal frequency. One or more diagnostic features were detected by transesophageal echocardiography in 88% of patients with endocarditis: vegetations (approximately with equal frequency on the mitral and the aortic, rarely on the tricuspid valve) were detected in 75% and/or abscesses in 22% of them. Only one patient in the control group with a history of endocarditis had vegetation-like structures on the aortic valve. Transesophageal echocardiography was on the average performed on the seventh day of hospitalization. With the exception of one female patient, all had been treated with antibiotics. Valve replacement was recommended in 65% of the patients with endocarditis and was in 59% also performed (single or double valve replacement), most frequently because of aortic or mitral regurgitation. The other patients were treated with antibiotics. Transesophageal echocardiography thus detects vegetations and abscesses in endocarditis frequently, findings that influence treatment in a significant manner.

Adult↗

PubMed1994

MAGE-1 gene product is a cytoplasmic protein.

MAGE-1 gene encodes a human melanoma antigen, recognized by syngeneic cytotoxic T lymphocytes (CTL). MAGE-1 transcripts are also detectable in breast cancers, in non-small-cell lung carcinomas and in central nervous system tumors. In order to identify, in cellular preparations, the protein encompassing the antigenic peptide, we generated a panel of monoclonal antibodies (MAbs) against the MAGE-1 gene product by using, as immunogen, a full-length recombinant preparation (rMAGE-1), obtained through expression cloning of the relevant gene in E. coli. Four reagents were obtained recognizing both rMAGE-1 and the 46-kDa native protein in cell lines expressing MAGE-1 mRNA. No positivity could be detected in MAGE-1-mRNA-negative melanoma lines. No surface labelling of MAGE-1-positive cell lines could be observed. In contrast, on permeabilization of MZ2 melanoma cells, all 4 MAbs induced efficient staining, as detected by cytofluorography. Fluorescence microscopy shows that MAGE-1 gene product is a cytoplasmic protein clustered in paranuclear organelle-like structures. Thus, MAGE-1 protein location closely resembles that of P91A and P198 murine-tumor antigens.

Animals↗

PubMed1994

[Surgery in pheochromocytoma. 12-year development].

Hypertension (sustained and/or paroxysmal) is the most common symptom of pheochromocytoma. Correct evaluation nearly always leads to the diagnosis, and complete surgical excision is the definitive treatment. Our study deals with preoperative evaluation, operative morbidity and long-term results after surgery. During the 12-year period (1981-1992) 31 patients (48 years, 25-85) with pheochromocytoma were treated at our institution. The selected preoperative management was reviewed on the basis of patient records. Long-term follow-up was carried out consecutively and personally. All patients received alpha-adrenergic blockers to achieve preoperative normotension and volume repletion. All resections were performed by laparotomy; two of the early cases were extended into the chest. 27 of 29 patients (93.1%) had elevated levels of urinary vanillylmandelic acid; a similar sensitivity was noted in the urinary catecholamines (24/26 = 92.3%). In anatomical localization CT-scan was accurate in 100% (27/27). MIGB scintigraphy was of help in 21 of 23 (91.3%) and, in the most recent patients, MRI was helpful in 3 of 3 (100%). No patient died during the first 30 days after surgery. One patient developed an anaphylactic reaction after cephalosporin administration (morbidity: 3.2%). 2 patients (6.4%) with malignant pheochromocytoma developed a recurrent tumor during the follow-up period. Resection of pheochromocytomas by laparotomy has a low perioperative morbidity and mortality after standardized preparation. A positive MIBG scintigraphy allows extra-adrenal localization. Better resolution and multiplanar imaging are the advantages of MRI. Perioperative management and new imaging techniques--eliminating arteriography and venous sampling among others--have simplified surgical treatment and improved its safety.

Adrenal Gland Neoplasms↗

PubMed1994

[Tumor-immune system interaction in renal cell carcinoma and melanomas. Cytokine transcription in tumors at the time of surgical resection].

Tumor infiltrating lymphocytes (TIL) of many tumors express surface activation markers. An antigen driven stimulation of T-lymphocytes is expected to induce not only cell membrane activation molecules but also a unique pattern of cytokine gene transcripts. These cytokines are relevant modulators and potent effectors of immune responses, and therefore play a crucial role in tumor-immune system interaction. The gene transcription of interleukin(IL)-2, IL-4, IL-7, IL-10 and interferon(IFN)-gamma of lymphocyte infiltrated, freshly excised tumor specimens from 10 renal cell carcinomas and 6 melanomas were investigated by reverse polymerase chain reaction (PCR) technique. Autologous, peripheral mononuclear cells (PBMC) and healthy tissue of the affected organs served as controls. In all samples the transcription of the beta-actin gene as a methodological control turned out to be positive. In contrast, no cytokine gene transcription was detected in healthy tissue specimens and PBMC. IL-2 transcripts were detectable in no melanomas but in half of the renal tumor samples. IL-10 never transcribed in melanomas but was positive in 7 out of 10 renal cell carcinomas. In only 2 respectively 1 of the resected tissue probes was transcription of IL-4 and IFN-gamma detected. IL-7 was positive in 1 melanoma and in 6 urological neoplasias. The most impressive fact is the frequent transcription of the inhibiting factor IL-10 in renal cell carcinomas (7/10). This pattern of cytokine gene transcription may explain functional deficits of TIL.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1994

The pattern of cytokine gene expression in freshly excised human metastatic melanoma suggests a state of reversible anergy of tumor-infiltrating lymphocytes.

Expression of an extended panel of cytokine genes was investigated by reverse polymerase chain reaction (PCR) in 10 freshly excised melanoma metastases infiltrated by lymphocytes (TIL). cDNA encoding for CD3-delta and tyrosinase could be amplified in all samples, confirming the presence of T lymphocytes and melanoma cells. Cytokine genes possibly transcribed by both cell types, such as GM-CSF, IL-6 and IL-10 could be amplified from 5, 2 and 2 samples respectively. In contrast, IL-1 beta and TNF-alpha mRNA were never detectable, IL-1 alpha, IL-3 and IL-7 mRNA could be observed only in one case each. Transcripts encoding for TGF-beta 1 were observed in 8 samples, while TGF-beta 2 and 3 mRNA were detectable in only 2 specimens. mRNA encoding for cytokine genes typically transcribed by antigen-stimulated T lymphocytes, such as IL-2, IL-4 and IFN-gamma were rarely or never detectable (none, none and 1 of the samples respectively). In one case, where no cytokine gene transcription was detectable at the time of surgery, we addressed the question of the antigenicity of the tumor and of the functional competence of TIL. A primary tumor cell line was generated and cultured TIL were induced to transcribe IL-2 and IFN-gamma genes by incubation with the autologous irradiated tumor cell line, but not with autologous EBV-transformed cells. In these conditions, tumor-specific cytotoxic T lymphocytes (CTL) could be generated only after 3 weekly re-stimulations. In contrast, if autologous irradiated EBV-transformed cells were added to the cultures, specific CTL could be detected after one single tumor stimulation. Thus, signs of active responsiveness in terms of lymphokine gene mRNA are seldom detectable in melanoma metastases. Tumor-specific responses, however, including IL-2 and IFN-gamma gene expression and generation of CTL can be produced in vitro from specimens in which no cytokine gene mRNA is detectable ex vivo.

Adult↗

PubMed1994

[Diagnosis and significance of antinuclear antibodies].

Explore the source record for details and available documents.

Animals↗

PubMed1994

Effects of allopurinol on in vivo suppression of arthritis in mice and ex vivo modulation of phagocytic production of oxygen radicals in whole human blood.

Recently, we demonstrated elevated levels of xanthine oxidase in serum of patients with various inflammatory and autoimmune rheumatic diseases. The present study reports the antiarthritic efficacy of the xanthine oxidase inhibitor and immunosuppressant allopurinol in DBA/1xB10A(4r) mice suffering from peroxochromate-induced arthritis. A profound dose-dependent suppression of arthritis was noted (P < 0.001). The ED50 was 80 +/- 14 mumol/kg/day. The arthritis index correlated positively to the phagocytic production of oxygen radicals (r2 > 0.672) and negatively to the concentrations of allopurinol (r2 = 0.915). Ex vivo, allopurinol and various conventional antirheumatic drugs were screened for the inhibition of 12-O-tetradecanoylphorbol-13-acetate-stimulated whole human blood chemiluminescence. The concentrations of antirheumatic drugs required to inhibit the chemiluminescence by 50% were compared to the therapeutic doses administered to rheumatic patients. While D-penicillamine and cis-platinum(II) increased the phagocytic generation of superoxide, nonsteroidal antiinflammatory drugs (NSAIDs), steroids, and slow-acting antirheumatic drugs (SAARDs) inhibited the whole blood chemiluminescence in a dose-dependent manner. Therapeutic doses of NSAIDs, SAARDs, or steroids inhibited the phagocytic generation of reactive oxygen species by 10-50%. In addition to well-known mechanisms of action of NSAIDs and SAARDs, our results support the hypothesis that most common anti-rheumatic drugs act also by modulating the levels of reactive oxygen species, which serve important mediator and signal transduction functions in inflammatory and autoimmune diseases. Pharmacologically safe antioxidants like allopurinol, which simultaneously modify the oxidative burst of phagocytes, inhibit xanthine oxidase, and display immunosuppressive effects may well be suited to control the consequences of chronic phagocytic hyperreactivity in rheumatic patients.

Allopurinol↗