Incorporation of simian virus 5 fusion protein into murine leukemia virus particles and its effect on the co-incorporation of retroviral envelope glycoproteins

We describe the generation of murine leukemia virus (MLV) virus particles carrying the paramyxovirus fusion protein F from simian virus 5 (SV5-F). This glycoprotein was expressed in cells providing Moloney MLV (MoMLV) Gag and Pol proteins and a lacZ retroviral vector. SV5-F was correctly expressed, processed, and efficiently incorporated into retroviral particles. SV5-F-bearing retroviruses were not infectious although a weak binding to primate and rodent cells could be detected and SV5-F could mediate cell to cell fusion. We then co-expressed the SV5-F glycoprotein in retroviral particles with chimeric and wild-type MoMLV envelope glycoproteins. Our results show that F strongly inhibited infection via the retroviral envelopes although the mechanism of inhibition was different depending on the retroviral envelope used.     

Effect of isoniazid, a haem inhibitor, on globin chain synthesis in reticulocytes from non-thalassaemic and beta thalassaemic subjects.

The effect of isonicotinic acid hydrazide (INH), a potent haem inhibitor, on globin chain synthesis was studied in reticulocytes from the following groups of patients: four non-thalassaemic patients (group i); five beta thalassaemia heterozygotes (group ii); three Hb S/beta thalassaemia heterozygotes (group iii); and two additional patients--one with homozygous beta thalassaemia and the other with thalassaemia intermedia (group iv). This was done to determine whether haem inhibitors depress alpha globin chain synthesis. The progressive increase of INH concentration (10-40 mmol l-1) in reticulocytes from a beta thalassaemia heterozygote resulted in a remarkable decrease of the alpha and beta chain synthesis, ranging from 80% to 97% and from 74% to 96% of control values, respectively, and in a gradual drop of alpha:beta ratio from 1.87 to 1.38. Furthermore, in the samples incubated with 40 mmol l-1 INH, a pronounced inhibition of globin chain synthesis 77 (19%) for alpha chain and 67 (27%) for beta or beta S chain) and a substantial drop of the alpha:beta or beta S ratio in samples with INH (median 1.16) compared with that in samples without INH (median 1.70) were observed. The inhibitory effect of INH was significantly or completely corrected by adding exogenous haem. It is suggested that haem inhibition and the resulting preferential diminution of alpha chain synthesis could provide a new approach to the treatment of homozygous beta thalassaemia with an excess of detrimental free alpha chain in erythroid cells.     

Reproducible lactulose hydrogen breath test as a measure of mouth-to-cecum transit time

Breath hydrogen monitoring after oral lactulose syrup is a conventional measure of mouth-to-cecum transit time (MCTT), but its reproducibility has been questioned. We compared the reproducibility of five measurements of MCTT after a conventional breakfast (380 kcal) taken with tea containing 20 g lactulose to five measurements of MCTT after 20 g lactulose in water in eight normal volunteers. Individual mean breakfast transit time was not significantly different from lactulose transit time in each of the seven subjects, but one had a breakfast transit time of 151±15 min and a lactulose transit time of 86±22 minutes (¯X ±sd, P<0.001). The coefficient of variation of breakfast transit time (11.6±5.3%, range: 6.9–24.2%) was less than that of lactulose transit time (30.7±7.8%, range: 22.1–50.0%, P<0.001). In a second set of experiments, the liquid phase marker ( ^99m Technetiumdiethylene triamine pentaacetic acid) emptied from the stomach more rapidly after the lactulose solution (T1/2 16.3±5.4) than after the breakfast (33.9±10.9 min, P<0.01) and MCTT was shorter after lactulose (77±32 vs 104±40 min, trespectively, P<0.05). There was no correlation between MCTT of lactulose and breakfast and between half-time gastric emptying and MCTT of either lactulose or breakfast. We conclude that the ingestion of inert lactulose induces an abnormally rapid MCTT and that breakfast MCTT is a much more reproducible investigation and should be employed in studies requiring serial measurements.     

Uroplakin Ib gene transcription in urothelial tumor cells is regulated by CpG methylation

Uroplakin Ib is a structural protein on the surface of urothelial cells. Levels of uroplakin Ib mRNA are dramatically reduced or absent in many transitional cell carcinomas, but the molecular mechanisms responsible remain undetermined. Previously, we showed that loss of uroplakin Ib expression correlated with CpG methylation of Sp1/NFkappaB-binding motifs within the proximal promoter. In this study, we show that reporter activity was completely blocked by the methylation of three CpG pairs in this promoter region. Gel shift analysis using purified proteins or nuclear extracts showed that Sp1 and NFkappaB bound to motifs encompassing two of the three CpG pairs. Interestingly, the methylation of these two CpG sites did not prevent the binding of proteins to the promoter in gel shift analyses. Additionally, mutation of these two CpGs did not affect reporter activity, but mutation of 6-bp fragment spanning each CpG partially inhibited reporter activity, suggesting that these sites were functional. A requirement for both Sp1 and NFkappaB in regulating reporter activity was confirmed in transfection experiments using plasmids expressing individual proteins. Our data suggest that the methylation of specific CpG sites can silence the uroplakin Ib promoter, at least in part, by blocking the binding of Sp1 and NFkappaB, although other factors may be involved.     

Malignant myoepithelioma arising from recurrent pleomorphic adenoma of the soft palate

Malignant myoepithelioma is a rare salivary gland neoplasm that can arise either de novo or within a pre-existing pleomorphic adenoma. We report a case of malignant myoepithelioma arising from a pleomorphic adenoma of the soft palate. A 70-year-old woman presented in our department with a very large tumor of the soft palate. The patient had a history of a pleomorphic adenoma at the same location that was surgically removed 2 years ago. A second operation, with intraoral excision of the tumor was performed. Histological examination of the recurrence revealed a malignant spindle cell neoplasm with an infiltrative growth pattern and a high mitotic rate. Immunohistochemical investigation confirmed the diagnosis of a low-grade carcinoma of myoepithelial origin with free surgical margins. The patient remains free of disease for more than twelve months after the end of treatment.     

Long-term results of a new stapedotomy prosthesis

The long-term results of a new, home-made stapedotomy prosthesis are presented. The innovations of this prosthesis concern both portions, design for attachment to the incus and shaft. Our series include a total number of 42 individuals with stapes fixation, operated using the new prosthesis, with a medical follow-up period between eighteen months and five years. Our innovations offer a proper and safe insertion of the prosthesis into the oval window associated with excellent manipulation and handling. These advantages could be of major importance for younger surgeons and trainees. Postoperative hearing results are similar to those achieved with other commercially available pistons.     

Radiation exposure to cardiologists performing interventional cardiology procedures

Medical doctors, who practice interventional cardiology, receive a noticeable radiation dose. In this study, we measured the radiation dose to 9 cardiologists during 144 procedures (72 coronary angiographies and 70 percutaneus translumined coronary angioplasties) in two Greek hospitals. Absorbed doses were measured with TLD placed underneath and over the lead apron at the thyroid protective collar. Based on these measurements, the effective dose was calculated using the Niklason method. In addition, dose area product (DAP) was registered. The effective doses, E, were normalised to the total DAP measured in each procedure, producing the E/DAP index. The mean effective dose values were found to be in the range of 1.2-2.7 microSv while the mean E/DAP values are in the range of 0.010-0.035 microSv/Gycm2. The dependence of dose to the X-ray equipment, the exposure parameters and the technique of the cardiologist were examined. Taking under consideration the laboratories' annual workload, the maximum annual dose was estimated to be 1.9 and 2.8 mSv in the two hospitals.     

A contribution to the establishment of diagnostic reference levels in CT

CT has become the major source of population exposure to diagnostic X-rays. CT dose index (CTDI) and dose-length product (DLP) have been proposed as the appropriate dose quantities for the establishment of diagnostic reference levels for optimizing patient exposure. Dose measurements on 27 CT scanners in Northern Greece involving six routine CT examinations have been performed in order to compare their performance with the currently proposed European reference dose values and to produce a preliminary set of data for the establishment of local diagnostic reference levels. All measurements were performed using a pencil shaped ionization chamber introduced into polymethyl methacrylate cylindrical head and body phantoms. The results revealed significant discrepancies in dose values among the CT scanners, which can be mainly attributed to variations in the examination protocols and the different kinds of scanners. Significant overdosing compared with the European reference levels has not been observed, with the exception of the routine head examination, where 47% of the scanners exceeded the corresponding CTDI(w) value. CT scans in the trunk region result in the higher effective doses, which can reach estimated maximal values of the order of 15 mSv.