PET for evaluation of differential myocardial perfusion dynamics after VEGF gene therapy and laser therapy in end-stage coronary artery disease.

The purpose of this study was to appraise the value of PET in the assessment of the effect of supposedly proangiogenic new therapies such as gene therapy with vascular endothelial growth factor (VEGF) gene and endomyocardial laser therapy. METHODS: Thirty-five patients with end-stage coronary artery disease and class III (Canadian Cardiovascular Society) angina were included. Myocardial ischemia was evaluated with dipyridamole PET scanning and exercise tolerance with bicycle ergometry. Ten patients were treated with naked plasmid DNA encoding for human VEGF165 (VEGF) and 12 patients were treated with laser therapy (direct myocardial revascularization [DMR]) using an electromechanical mapping system. Thirteen patients were treated with standard medical therapy (control). RESULTS: In both active treatment groups, angina was reduced in most subjects, except in 2 VEGF and 5 DMR patients. In the control group, no improvement in anginal classification was found, except in 3 subjects. On the PET scan, solely in the VEGF group, the stress perfusion was significantly improved (from 57 +/- 33 to 81 +/- 55 mL/min/100 g; P = 0.031). Furthermore, in the VEGF group, the number of ischemic segments was reduced from 274 +/- 41 to 234 +/- 48 segments (P = 0.004) but not in the DMR group (from 209 +/- 43 to 215 +/- 52 segments) or in the control group (from 218 +/- 18 to 213 +/- 28 segments). Bicycle exercise duration showed slight nonsignificant changes in the VEGF group (from 3.6 +/- 2.0 to 4.6 +/- 2.1 min), in the DMR group (from 5.1 +/- 1.5 to 4.7 +/- 1.3 min), and in the control group (from 3.3 +/- 1.8 to 3.5 +/- 2.3 min). CONCLUSION: PET showed that intramyocardial gene therapy with the human VEGF165 gene in contrast to laser DMR treatment effectively reduces myocardial ischemia.     

Diagnosis of fetal rubella infection by nucleic acid hybridization

The efficacy of nucleic acid hybridization for the diagnosis of rubella infection in experimental and clinical materials was compared with immunoblot and virus isolation techniques. Our results showed that nucleic acid hybridization is specific and rapid but gives false-negative results when compared with conventional virus isolation in some experimental although not in clinical materials so far examined. For this reason, a failure to demonstrate rubella virus in fetal specimens by this method alone cannot yet be taken as a sole criterion for ruling out fetal rubella infection.     

Functional characteristics of coronary vasomotor function following intramyocardial gene therapy with naked DNA encoding for vascular endothelial growth factor165.

Vascular endothelial growth factor (VEGF) is a potent angiogenic factor. VEGF gene therapy improves perfusion of ischemic myocardium in experimental models and possibly in patients with end-stage coronary artery disease. In addition to its proliferative and migratory effect on endothelial cells, it also activates and up-regulates endothelial nitric oxide synthase (eNOS). Therefore, the authors investigated coronary endothelium-dependent vasodilatation in patients before and after VEGF gene therapy. The effect of intracoronary acetylcholine infusion on coronary diameter was assessed at baseline and after 3 months follow-up in patients with end-stage coronary artery disease treated with VEGF gene and in controls scheduled for elective percutaneous transluminal coronary angioplasty (PTCA) (acetylcholine test at diagnostic angiography and before a subsequently scheduled PTCA). Five out of six VEGF patients experienced a reduction in anginal complaints. Angiographic evidence for improved collateral filling was evident in two out of six patients. The vasoconstrictive response to acetylcholine was partly converted into dilatation. In contrast, the acetylcholine response in control patients remained vasoconstrictive. In conclusion, VEGF gene therapy has an important beneficial effect on the functional characteristics of the myocardial vascular network. Therefore, this therapy can potentially play an important role in all stages of the atherosclerotic process.     

Rapid detection of respiratory viruses using mixtures of monoclonal antibodies on shell vial cultures.

Eleven hundred and thirty-three clinical specimens submitted to the laboratory for diagnosis of respiratory virus infections were tested by direct immunofluorescence (DIF) for respiratory syncytial virus (RSV), by shell vial culture, and by conventional cell culture. The shell vial cultures were stained with 8 different monoclonal antibodies both 1 day and 3-7 days after inoculation. In order to limit the cost and the workload, mixtures of monoclonal antibodies were used. Coverslips with HEp-2 cells were incubated with a mixture of FITC-labeled monoclonal antibody to RSV and nonlabeled monoclonal antibody to adenovirus. When no RSV positive IF staining was observed after the first incubation step, the same coverslip was incubated once more with FITC-labeled anti-mouse antibody. A positive reaction at this stage indicated the presence of adenovirus. Similarly, cultures of tertiary monkey kidney cells were investigated with a mixture of two FITC-labeled monoclonals to the influenza viruses A and B and three nonlabeled monoclonals to the parainfluenza viruses 1, 2 and 3. If influenza virus or parainfluenza virus was detected, the exact type was determined by staining different parts of a duplicate coverslip. Shell vial cultures for cytomegalovirus (CMV) were always performed separately on human embryonic lung fibroblasts. Using this approach, we detected RSV (n = 248), CMV (n = 42), parainfluenza virus (n = 31), influenza virus (n = 28), and adenovirus (n = 6), in most cases after only one day of culture. For RSV, the sensitivity of the shell vial method was too low (74%) to allow omission of DIF (sensitivity 95%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Reproducible marginal gap measurements of partial ceramic crowns

Aim of this research was to test the hypothesis that marginal gap measurements by a digital microscopic image processing computer are reproducible. Light microscopic images of the margins of Artglass partial crowns on stone dies were obtained with a 3CCD color video camera, digitized and processed. Ten measuring points were painted on a 0.5 mm section of the finish line of the preparation. The opposite corresponding measuring points on the margin of the partial crown were painted as well. The distance (marginal gap) between two corresponding points was painted and calculated by the computer. Reproducibility (precision) is reflected in the ability to reproduce the same measurement results in repeated measurements (n = 5). The reproducibility error was expressed as the coefficient of variation (CV) in percent (%). The localization of the corresponding measurement points and the calculation of the marginal gap were performed without errors. For a marginal gap between 15-50 microns the average measurement error was 3% or 0.45-1.5 microns. It was concluded that the measurement of the gap between two points by the digital image processing computer was reproducible. The measurement error was neglectable given that a clinically acceptable marginal gap is 100 microns.     

The effect of ceftriaxone on the anaerobic bacterial flora and the bacterial enzymatic activity in the intestinal tract

Summary The normal flora of the intestinal tract, mainly consisting of anaerobic bacteria, protects the host against colonization by pathogenic microorganisms. Antimicrobial treatment with ceftriaxone may influence the colonic microflora and as a consequence, the protective effect. Ten healthy volunteers received 1 g of ceftriaxone intramuscularly for five days. This resulted in a significant decrease (p<0.05) of the mean cultural counts (± SEM) of total anaerobes from 10.67 (0.11) (prior to treatment) to 9.02 (0.45) and 8.97 (0.46) at days 3 and 5, respectively (during treatment). After treatment (days 10 and 15–19), the cultural counts of anaerobes returned to 10.17 (0.16) and 10.44 (0.18), respectively. Bacterial enzymes may serve as an indicator of protective microflora. - aspartylpeptidase and deoxycholate hydrolase activity was determined in faecal supernatants of the volunteers and compared with anaerobic culturing. Both enzymatic activities show a significant correlation with the total number of anaerobes present at day 3 of ceftriaxone treatment. At day 5 and 8 only -aspartylpeptidase showed significant correlations with cultural counts of total anaerobes,Bacteroides spp. or bifidobacteria. At day 15 to 19 (ten to 14 days after treatment) -aspartylpeptidase showed only a significant correlation with the number ofBacteroides spp. This indicates that changes in the indigenous bacterial flora during and shortly after treatment with ceftriaxone can be monitored by determination of -aspartylpeptidase. Recovery of the intestinal flora is difficult to assess in this manner.

---

Einfluß von Ceftriaxon auf die anaerobe Flora und die bakterielle Enzymaktivität im Intestinaltrakt

Zusammenfassung Die normale Flora des Intestinaltraktes besteht vorweigend aus anaeroben Bakterien und schützt den Wirt gegen eine Kolonisation durch pathogene Mikroorganismen. Eine antimikrobielle Therapie mit Ceftriaxon kann die Mikroflora des Dickdarms beeinträchtigen und damit auch deren protektiven Effekt. Zehn gesunde Probanden erhielten fünf Tage lang 1 g Ceftriaxon intramuskulär appliziert. Dies führte zu einer signifikanten Abnahme der mittleren Koloniebildnerzahlen von 10,67 (SEM ± 0,11) vor Applikation auf 9,02 (± 0,45) nach drei und auf 8,97 (± 0,46) nach fünf Tagen (p<0,05). Nach zehn und 15 bis 19 Tagen im Anschluß an die Antibiotikagabe kehrten die Anaerobier-Koloniebildnerzahlen auf 10,17 (± 0,16) bzw. 10,44 (± 0,18) zurück. Bakterienenzyme können als Indikator für die protektive Mikroflora dienen. In Überständen von Stuhlproben der Probanden wurden -Aspartylpeptidase und Desoxycholat-Hydrolase bestimmt und mit den Anaerobier-Kulturen verglichen. Zwischen den Aktivitäten beider Enzyme und der am Tag 3 gemessenen Anaerobier-Gesamtzahl fand sich eine signifikante Korrelation. Am Tag 5 und Tag 8 zeigte nur die -Aspartylpeptidase eine signifikante Korrelation mit den Gesamt-Kolonie-bildnerzahlen der Anaerobier sowie mit den Zahlen vonBacteroides spp. oder Bifidobakterien. An den Tagen 15 bis 19 (zehn bis 14 Tage nach Antibiotikagabe) bestand nur zwischen der Zahl vonBacteroides spp. und -Aspartylpeptidase eine signifikante Korrelation. Nach Behandlung mit Ceftriaxon lassen sich folglich Veränderungen der bakteriellen Flora kurzfristig durch Bestimmung der -Aspartylpeptidase erfassen, weniger gut aber die Erholung der Darmflora.

     

Different Recurrence Pattern After Neoadjuvant Chemoradiotherapy Compared to Surgery Alone in Esophageal Cancer Patients

Purpose

To evaluate the rate and pattern of recurrences after neoadjuvant chemoradiotherapy (CRT) in esophageal cancer patients.

Methods

We described survival and differences in recurrences from a single center between neoadjuvant CRT (carboplatin/paclitaxel and 41.4 Gy) and surgery alone for the period 2000–2011. To reduce bias, we performed a propensity score matched analysis.

Results

A total of 204 patients were analyzed, 75 treated with neoadjuvant CRT and 129 with surgery alone. The pathologic response to neoadjuvant CRT was 69 % with a complete response rate of 25 %. After matching, baseline characteristics between the groups (both n = 75) were equally distributed. The 3- and 5-year disease-free survival was 53 and 42 % in the neoadjuvant CRT group compared with 24 and 18 % in the surgery-alone group (P = 0.011). After 3 and 5 years’ CRT, patients had an estimated locoregional recurrence-free survival of 83 and 73 % compared with 52 and 49 % in the surgery-alone group (P = 0.015). The distant recurrence-free survival was comparable in both groups. Locoregional recurrences were located less in the paraesophageal lymph nodes in the CRT group than in the surgery-alone group, 9 versus 21 %, respectively (P = 0.041). With respect to differences in distant recurrences, we observed more skeletal recurrences in the surgery-alone group compared to CRT, 12 versus 1 % (P = 0.009).

Conclusions

The neoadjuvant CRT regimen we used offers a significant improvement in outcome, with a different recurrence pattern compared with surgery alone. This effect is probably due to both the pathologic complete response and eradication of micrometastases in CRT group.     

Evaluation of [18F]FHPG as PET tracer for HSVtk gene expression

In rats, the relationship between [(18)F]FHPG accumulation and HSVtk expression was studied with PET and autoradiography. [(18)F]FHPG distribution closely corresponded with HSVtk immunohistochemical staining. ROI analysis of tracer uptake 2 hours p.i. and Patlak graphical analysis were applied to quantify the PET data. For both analysis methods, the [(18)F]FHPG PET signal correlated well with the fraction of HSVtk expressing cells implanted, but showed a plateau when plotted against HSVtk protein levels. This might be due to rate limiting [(18)F]FHPG membrane transport.