Different microvascular endothelial cell phenotypes exhibit different class I and II antigens under interferon-gamma.
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Animals↗
Biomedical subjects
G Bein
Publications and source records attributed to G Bein.
Cytomegalovirus transcripts in peripheral blood leukocytes of actively infected transplant patients detected by reverse transcription-polymerase chain reaction.
In an effort to examine human cytomegalovirus (CMV) infections on the transcript level in vivo, a reverse transcription-polymerase chain reaction (RT-PCR) for the detection of CMV mRNA in human peripheral blood leukocytes (PBL) was developed. Oligonucleotide primers were derived from the major immediate-early (MIE) and the pp150 genes of CMV, which allowed the exact differentiation between viral mRNA and DNA. With these primers, 8 renal transplant patients who revealed some evidence of active CMV infection were investigated. CMV-specific transcripts were found in 5 patients, all of them presenting MIE mRNA. pp150 transcripts were demonstrated in only one symptomatic patient who showed RNAemia for several weeks. These findings suggest that during active infection, CMV replicates in PBL and, furthermore, that a detailed analysis of mRNA patterns may make it possible to identify those patients at highest risk of developing symptomatic infection.
Antigens, Surface↗
Arterioportal fistula in infancy.
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Angiography↗
The long persistence of CMV DNA in the blood of renal transplant patients after recovery from CMV infection.
A total of 30-50% of all renal transplant recipients undergo infections caused by human cytomegalovirus. With the introduction of ganciclovir and foscarnet for specific antiviral therapy there is an increasing demand for diagnostic tools that allow the early and rapid identification of CMV as the causative agent of the observed disease. We and others previously showed the direct detection of pp65 antigen in peripheral blood leukocytes to be an excellent marker for active cytomegalovirus infection. In order to establish whether the detection of CMV DNA by the polymerase chain reaction (PCR) supplies further information in this regard, we compared both methods. In 41 renal transplant patients the PCR assay yielded a sensitivity of 100% compared with 87.5% of the antigenemia assay. Specificities reached 67% and 92.5%, respectively. In 5 patients without both serological signs of infection and antigenemia, CMV DNA was also found. The duration of CMV DNA detection in PBL during active infection was significantly longer than antigenemia. Even after successful treatment of symptomatic CMV disease, DNA was present for a period of weeks without any relapse of disease. In contrast, antigenemia disappeared after antiviral therapy and reappeared only in one patient with relapse of CMV disease. We conclude that PCR offers no advantages over antigen detection in monitoring for CMV infections after renal transplantation.
Base Sequence↗
HLA DNA preorgan retrieval donor typing in renal transplantation.
For the first time, a DNA technique was used in clinical practice for pre-organ donor retrieval typing in renal transplantation. The method is based on the polymerase chain reaction with nested sequence-specific pairs (nested PCR-SSP) for all serological HLA-DR specificities of DR1-DR18. A panel of reference-typed individuals (n = 101) was investigated in a blind quality control study and the results revealed a sensitivity of 99.5% and a specificity of 100%. Semi-automation of PCR-SSP provided rapid, accurate and reliable typing results demonstrating that this DNA test is most suitable for replacing the error-prone HLA-DR serological technique. Twenty-seven consecutive cadaveric organ donors have been successfully typed so far. The results were available within less than 3 h after blood sampling and in all cases prior to the retrieval of organs. In conclusion, the introduction of this accurate DNA typing in HLA matching programs may significantly improve the graft survival rate in renal transplantation.
Blood Grouping and Crossmatching↗
[Diagnosis of cytomegalovirus infections in immunosuppressed patients: the role of PCR].
The well-calculated and early application of specific antiviral drugs in cytomegalovirus (CMV) infections depends on sufficient virological diagnosis. Many authors prefer the detection of CMV DNA by the polymerase chain reaction (PCR) for this purpose. In our study in 41 renal transplant patients PCR yielded no additional information of clinical relevance compared to the already established pp65 antigenemia assay. However, neither of the two techniques allowed the prediction of symptomatic infections as also asymptomatic infections yielded positive results. In contrast, CMV mRNA was demonstrated by PCR mainly in symptomatic infections. We conclude, that mRNA amplification possibly is able to predict symptomatic CMV infections and will thereby allow the well-calculated application of e.g. ganciclovir.
Cytomegalovirus Infections↗
Beta2-microglobulinuria as an early sign of cytomegalovirus infection following renal transplantation.
The frequency of cytomegalovirus infection was studied in a prospective study of 106 kidney recipients. The detection of cytomegalovirus-immediate-early-antigen and cytomegalovirus-immunoglobulin (IgM) antibodies in serum was used as the reference method and showed that 23.6% (25/106) of all patients were infected. In addition, four urinary proteins (IgG and transferrin as glomerular markers and alpha1-microglobulin and beta2-microglobulin as tubular markers) were quantitatively measured in 24-h urine samples from all of the patients using an immunoluminometric assay (ILMA). In all cytomegalovirus infection cases a pronounced but isolated increase of urinary beta2-microglobulin excretion was observed. In 20 of 25 infected patients, the beta2-microglobulinuria occurred 1-21 days (median 5.0) earlier than the appearance of the cytomegalovirus-immediate-early-antigen in blood. Thus, it can be seen that the quantitative measurement of beta2-microglobulin in urine is useful for the early detection of cytomegalovirus infection following renal transplantation.
Biomarkers↗
Failure to detect human cytomegalovirus DNA in peripheral blood leukocytes of healthy blood donors by the polymerase chain reaction.
The transmission of cytomegalovirus (CMV) by blood transfusion may have a major effect on certain immunocompromised patients. To protect susceptible blood recipients from infection, it is advisable to use blood components from CMV-seronegative donors. However, serologic tests are not capable of indicating which blood component actually harbors infectious virus and can transfer it to the recipient. Therefore, a sensitive method is needed for the detection of the virus itself. There have been three reports on the detection of CMV in healthy volunteer blood donors by the polymerase chain reaction (PCR). CMV DNA was found in all seropositive and most seronegative blood donors. However, many other authors have failed to confirm these data. A highly sensitive and specific PCR assay was developed for the detection of CMV DNA in peripheral blood leukocytes. With this protocol, blood samples from 116 volunteer blood donors were investigated. None of these samples proved to be positive for CMV DNA. In contrast, CMV DNA was detected in 10 of 10 renal transplant patients early in the course of active CMV infection. It can be concluded that the CMV genome copy number in the peripheral blood leukocytes of healthy individuals is beyond the detection limit of current PCR technology.
Base Sequence↗
Rapid HLA-DRB1 genotyping by nested PCR amplification.
State of the art genotyping of HLA class II alleles with group-specific DNA amplification by the polymerase chain reaction (PCR) (1) and subsequent probing with sequence-specific oligonucleotides (2-4) is not suitable for typing cadaveric organ donors since the typing procedure takes far more than one working day. We designed specific oligonucleotide primer sets for nested PCR amplification which allowed typing for all serological HLA-DR specificities (DR1-DRw18) solely by the detection of amplified DNA in the reaction mixtures after agarose gel electrophoresis. Exon 2 of the DRB genes and a DRw52-group-specific part of DRB1 exon 2 was amplified directly from cell lysates without prior DNA extraction. The amplified DNA was subjected to a second round of amplification, which employed a set of 18 nested allele- or group-specific primer pairs. All alleles which have at least a single mismatched base at the terminal 3'-nucleotide of one primer were completely refractory to amplification. This assay is easy to perform and takes less than one working day to complete. Thus, this method may prove to be suitable for DNA typing of organ donors for prospective HLA-DR matching in renal transplantation.
Alleles↗
Detection and quantification of latently infected B lymphocytes in Epstein-Barr virus-seropositive, healthy individuals by polymerase chain reaction.
We designed a highly sensitive and specific polymerase chain reaction assay for the detection of Epstein-Barr virus (EBV)-related sequences in B-cell DNA of EBV-seropositive healthy individuals. By using this assay, we were able to amplify at least 10 copies of a plasmid containing the BamHI-W region, which is repeated up to 11 times within the EBV genome, in the presence of 1 microgram of EBV-negative DNA, indicating that one virus genome was detectable from 150,000 cells. In 15 of 16 tested individuals, EBV-related sequences were found frequently when the DNA from 10(6) B lymphocytes was examined and 1 microgram of DNA was used in each polymerase chain reaction. When the results of amplifying the diluted plasmid were used as a semiquantitative standard, the number of EBV genomes detected could be estimated to range between 50 and less than 1 from 10(6) B lymphocytes. The results of our study will provide the basis for further investigations of the characteristics of the latent carrier state in healthy EBV-seropositive individuals, such as the determination of the number of virus copies per cell and expression of antigens.
Antibodies, Viral↗
Molecular matching in renal transplantation.
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Alleles↗
Concordant Bell's palsy in monozygotic twins.
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Diseases in Twins↗
[Beta 2-microglobulinuria in cytomegalovirus infections following kidney transplantation].
In a prospective study the occurrence of cytomegalovirus (CMV) infection was diagnosed by demonstrating CMV-immediate early antigen (IEA) in the blood in 13 out of 68 (19%) patients who had undergone renal transplantation (27 women, 41 men, mean age 46.3 [21-64] years). Twenty-four hour urine samples were collected at the same time for quantitative determination of selected marker proteins by immunoluminometric assays (IgG and transferrin as glomerular markers, alpha 1-microglobulin and beta 2-microglobulin as tubular markers). In all 13 confirmed cases of CMV infection there was an isolated rise in urinary beta 2-microglobulin excretion amounting to more than three times the normal. In 11 of the 13 cases the beta 2-microglobulinuria was noted before the CMV-IEA test became positive. This suggests that quantitative assay of beta 2-microglobulin in the urine may be of value in the early diagnosis of CMV infection--a matter of some prognostic importance--and may hence offer additional support for therapeutic decisions.
Adult↗
The detection of human cytomegalovirus immediate early antigen in peripheral blood leucocytes.
Recently, Van der Bij et al. (1988) reported that active human cytomegalovirus (HCMV) infection could be diagnosed by the detection of HCMV immediate early antigen (IEA) directly in the peripheral blood leucocytes of renal transplant recipients. However, the indirect peroxidase technique used resulted in high background staining due to endogenous peroxidase activity and thus the detection of HCMV-IEA positive leucocytes, which are sometimes present in extremely low numbers, was not always reliable. In an attempt to solve this problem, we have evaluated the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique, immunogold-silver staining (IGSS), and several fixatives. Fixation with acetone: methanol 1:1 in conjunction with the APAAP technique proved to be the most successful method. In 155 blood samples obtained from 44 patients following renal transplantation and from three AIDS patients, the number of positive cells ranged between 1 and 700 out of 400,000 (median 2). In 23 samples from 11 patients (one AIDS patient) at least one positive cell was found. In this series there were no problems with the evaluation since strong positive signals were obtained without any background staining. We therefore recommend the use of this protocol for the rapid and reliable detection of HCMV-IEA in peripheral blood leucocytes.
Acquired Immunodeficiency Syndrome↗
[Surgical ligation of patent ductus arteriosus on the intensive care unit in small premature infants].
This paper summarizes the experience with ligation of persistent ductus arteriosus in prematures performed in the intensive care unit during the years 1986 to 1988. Records of 35 operated prematures on artificial ventilation were analyzed retrospectively. Management and organization of the operation are described. For comparison, records were analysed of 47 prematures, also artificially ventilated, who during the years 1978 to 1986 had to be transported to an operating theatre over a distance of 3 or 10 km for the ligation.
Ductus Arteriosus, Patent↗
The relevance of intraoperative pressure and oxygen saturation monitoring during pulmonary artery banding in infancy.
Different methods have been used to define an effective and tolerable degree of pulmonary artery banding (PAB) intraoperatively. As the hemodynamic response determines the results, a reduction of risk could be expected by evaluating several selected parameters and adjusting the band accordingly. Between 1981 and 1986, PAB was performed in 12 infants, three with complex cardiac lesions and nine with complete atrioventricular canal defects. Pressures in the distal pulmonary artery, in a systemic artery or the aorta, and in a central vein were monitored, and oxygen saturation measurements in both great arteries were taken before, during, and after the procedure. Adequate, complicated, intolerable, and contraindicated constriction were recognized and shown on characteristic pressure traces and oxygen saturation shifts. The measurements provided an indirect but reliable indication of the change in intracardiac shunting, and predicted whether the banding would be hemodynamically tolerated. Postoperatively, ten of the infants improved significantly, especially the seven infants previously in a low cardiac output state. One infant died on the day of surgery from acute right heart failure (8% hospital mortality). The results suggest that hemodynamically controlled PAB is an effective palliative treatment with a lower risk than had been previously reported.
Blood Pressure↗
Balloon valvuloplasty of pulmonary stenosis in infants and children--co-operative study of the German Society of Pediatric Cardiology.
Balloon valvuloplasty (BVP) of pulmonary stenosis (PST) was performed in 305 infants and children in 20 heart centres of the Federal Republic of Germany. 273 patients suffered from isolated PST. Their age ranged from 3 days to 18 years (mean = 5.85 +/- 4.82 years), their weight from 2.9 to 60 kg. The pressure gradient was measured invasively in the sedated or anaesthetized child, partly by CW Doppler. BVP was performed by mono- or trefoil catheters, mostly oversized. As a result, mean pressure gradient fell from 71.7 +/- 32 mmHg before to 31.8 +/- 25 mmHg immediately after BVP (P less than 0.0001). The gradient was reduced by less than 30% in 13% of the patients, by 30-49% in 23%, and by greater than or equal to 50% in 64%. Among 33 patients with a gradient reduction of less than 30%, nine were aged less than 1 year, nine had dysplastic valves, and nine had a short-term infundibular reaction and good late results. Gradient reduction was not improved with oversized balloons and was independent of the initial gradient. Follow-up for a mean 11.2 +/- 8 months showed a further slight improvement, probably due to regression of infundibular hypertrophy. In the whole group we observed infundibular reaction in 12%, serious complications with one late death in 4%. From the start, BVP proved to be a secure and effective tool for elimination of pulmonary stenosis.
Adolescent↗
Transposition of the great arteries with straddling tricuspid valve. Report of two rare cases with acquired subaortic stenosis after main pulmonary artery banding.
The clinical, hemodynamic, angiocardiographic, and postmortem findings of a previously unreported type of subaortic stenosis are presented in two patients who also had straddling tricuspid valve and transposition of the great arteries. The subaortic stenosis became apparent after banding of the main pulmonary artery and was due to hypertrophy of a probably abnormally positioned moderator band resulting in a double-chambered right ventricle. Fibrous tissue accumulation at the stenotic os infundibuli also contributed to the subaortic obstruction. Both patients had situs solitus of the atria. Patient 1 had ventricular inversion (L-loop ventricles) and atresia of the right-sided mitral valve. Patient 2 had normally positioned ventricles (D-loop) and two atrioventricular valves. The presence of a large left ventricle and a small right ventricle in the angiocardiogram led to the erroneous diagnosis of a single left ventricle with an infundibular outlet chamber in both patients. Consequently, the subaortic obstruction was thought preoperatively to be at the site of a restrictive bulboventricular foramen. Patient 1 died 36 hours after placement of a valved conduit from the left ventricle to the descending aorta. Patient 2 was operated on successfully and the surgical procedures performed are described.
Aortic Valve Stenosis↗