Plasma shield lasertripsy: in vitro studies

A technique for safer and more effective pulsed laser lithotripsy of urinary and biliary calculi was investigated in vitro. The technique involves enclosing the distal end of the laser delivery fiber in a "plasma shield." The plasma shield is a specially designed metal cap that serves to transfer the laser-induced mechanical impulse to the calculus while shielding surrounding tissue from direct laser exposure and thermal radiation. The metal cap also offers the advantage of effectively blunting the sharp fiber tip and improving its visualization under fluoroscopy. Plasma shield lithotripsy using a 200 micron quartz fiber inserted into a section of a modified 0.034 in. diameter stainless steel guide wire was tested in vitro on a variety of calculi and compared with results obtained using a 200 micron laser fiber applied directly. Calculi tested included cystine, struvite and calcium oxalate dihydrate urinary stones and pigmented cholesterol gallstones. The laser source was a flashlamp-pumped dye laser producing pulses of 1.2 microsecond duration and operated at a wavelength of 504 nm and pulse repetition frequency of 5 Hz. The results show that plasma shield lasertripsy is as effective as direct lasertripsy for fragmenting gallstones, struvite and calcium oxalate dihydrate calculi, is potentially safer, and can fragment cystine calculi which the pulsed dye laser applied directly cannot.     

Ensuring data quality in a multicenter clinical trial: remote site data entry, central coordination and feedback

In an ongoing multicenter clinical trial, "Treatment Strategies in Schizophrenia," the five participating sites have the capacity to perform a variety of tasks or study functions independently. These tasks include (a) verification of diagnostic eligibility through the use of computerized decision algorithms; (b) assignment of patients to treatment based on prognostic indicators using a computerized randomization algorithm; (c) entry of data into a microcomputer using a clinical trial data management system that performs simple range and missing data item checks; and (d) regular transfer of all data to the central coordinating team. The clinical trial data management system employed allows for both independent site functioning and assurance of consistency across sites. The integration of a variety of software outside the main data management system provides the central coordinators with the tools to monitor critical data as it is collected, as well as the capacity to assess the flow, quality, and uniformity of the ongoing trial.     

Protective actions of human recombinant basic fibroblast growth factor on MPTP-lesioned nigrostriatal dopamine neurons after intraventricular infusion

Basic fibroblast growth factor (bFGF, FGF-2) is a trophic factor for neurons and astrocytes and has recently been demonstrated in the vast majority of dopamine (DA) neurons of the ventral midbrain of the rat. Potential neuroprotective actions of FGF-2 in the l-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP) model have also been reported. The actions of the FGF-2 have now been further analyzed in a combined morphological and behavioural analysis in the MPTP model of the adult black mouse, using a continuous human recombinant FGF-2 (hrFGF-2) intraventricular (i.v.t.) administration in a heparin-containing (10 IU heparin/ml) mock cerebrospinal fluid (CSF) solution. Tyrosine hydroxylase (TH) immunocytochemistry in combination with computer assisted microdensitometry demonstrated a counteraction of the MPTP-induced disappearance of neostriatal TH-immunoreactive (ir) nerve terminals following the FGF-2 treatment. Unbiased estimates of the total number of nigral TH ir neurons, using stereological methods involving the optical disector (Olympus), showed that the MPTP-induced reduction in the number of nigral TH ir nerve cell bodies counterstained with cresyl violet (CV; by 56%) was partially counteracted by the FGF-2 treatment (by 26%). The behavioral analysis demonstrated an almost full recovery of the MPTP-induced reduction of the locomotor activity after FGF-2 treatment. This action was maintained also 1 week after cessation of treatment. The hrFGF-2 produced an astroglial reaction as determined in the lateral neostriatum and in the substantia nigra (SN) far from the site of the infusion, indicating that the growth factor may have reached these regions by diffusion to activate the astroglia. Immunocytochemistry revealed FGF-2 immunoreactivity (IR) in the nuclei of the astroglia cell population in the dorsomedial striatum and the microdensitometric and morphometric evaluation demonstrated an increase in the number, but not in the intensity, of these profiles on the cannulated side, suggesting the possibility that hrFGF-2 stimulates FGF-2 synthesis in astroglial cells with low endogenous FGF-2 IR. These results indicate that hrFGF-2, directly and/or indirectly via astroglia, upon i.v.t. infusion exerts trophic effects on the nigrostriatal DA system and may increase survival of nigrostriatal DA nerve cells exposed to the MPTP neurotoxin.     

Duplication of the upstream regulatory sequences increases the transformation potential of human papillomavirus type 11.

Infections with certain types of papillomaviruses, e.g., human papillomavirus type 16 (HPV-16), often progress to cancer. Malignant lesions associated with the closely related HPV-11 are extremely rare. Additionally, HPV-11 DNA, unlike HPV-16, does not normally transform cells in vitro. We determined that HPV-11 DNA was able to transform baby rat kidney cells in a ras-dependent focus assay when the upstream regulatory region (URR) was present in two copies. Addition of a second HPV-11 URR or an HPV-16 URR was equally effective and was position and orientation independent. The transformation was enhanced by dexamethasone. On passage the HPV-11 genome was not retained at a detectable level. This analysis supports isolated observations in vivo that duplications of regulatory sequences in HPV-11 increase the transformation potential of this virus type.     

Mapping of IgG subclass and T-cell epitopes on HIV proteins by synthetic peptides

Fifteen amino acid peptides, sequentially overlapping by 10 amino acids, were synthesized on the basis of the HTLV-III sequences of the gag and env proteins. They were used as antigens in IgG subclass ELISAs and T-cell stimulation assays. Sera and cells were obtained from 30 asymptomatic, HIV-infected homosexuals. In all subclasses reactivity was found to parts of the gag protein, while IgG1 dominated anti-env peptide responses. It was possible to delineate peptides showing restricted IgG subclass responses that were dominated by either IgG1, 2, 3 or 4. A negative correlation was generally observed between B-cell and T-cell reactivity, but a T-cell and B-cell co-operation was suggested by the response to two IgG1-restricted peptides. The IgG3-dominated epitopes were present in peptides previously known to be amphipathic and capable of T-cell stimulation. The analysis of subclass-restricted responses on the peptide level will assist the understanding of the subclass expression in vivo, since the peptide mapping approximates the delineation of a subclass-restricted response at the level of single epitopes.     

Simultaneous turnover of normal and dysfunctional C1 inhibitor as a probe of in vivo activation of C1 and contact activatable proteases.

Simultaneous turnover of normal and dysfunctional C1-inhibitor (C1-INH) was carried out in 10 normal subjects and 13 patients with rheumatoid arthritis as a measure of the in vivo activation of C1 and the contact activatable enzymes. In the first series of experiments, dysfunctional protein We was used in simultaneous turnover studies in five normal subjects and nine patients. The fractional catabolic rate of the dysfunctional C1-INH, We, (FCR(d)) was unchanged in both groups but the fractional catabolic rate of the normal C1-INH (FCR(n)) was faster in the patients compared to the controls, in particular patients with vasculitis. The enzyme-dependent catabolism defined as FCR(n-d) X concentration of C1-INH X plasma volume, was raised in the patient group, and correlated with disease activity score (r = 0.83, P less than 0.05). Neither FCR(n) nor FCR(d) was dependent on C1-INH concentration. The latter was higher in the patients (206 mg/l compared with 155 mg/l) indicating a very high synthetic rate in the patients (280.81 micrograms/kg/h compared with 179.77 micrograms/kg). In the second series of turnovers in six patients and five normal subjects, another dysfunctional C1-INH, at, was used. The FCR of C1-INH was slower than C1-INH (We) (1.88%/h compared with 2.7%/h). Enzyme-dependent catabolism of C1-INH in these patients were raised and also correlated with disease activity score (r = 0.82, P less than 0.05).     

Long term variation in beam symmetry as a function of gantry angle for a computer-controlled linear accelerator

Testing computer-controlled linear accelerators for patient safety and proper patient dose delivery requires that certain beam characteristics be monitored over an extended period of time. Computer-controlled conformal radiation therapy using asymmetric collimator jaw settings necessitates stable symmetric treatment beams. Long term beam symmetry measurements have been performed on a Philips SL20 dual energy computer-controlled linear accelerator. Symmetry in both the radial and transverse axis of each x-ray beam was monitored for eight gantry positions. These measurements were undertaken to determine the effectiveness of the SL20 beam steering system during dose delivery of 50 monitor units (MU) per field. Evaluation of the data shows that careful beam steering setup procedures result in x-ray beams in which fluctuations in symmetry as a function of gantry angle are within +/- 1.5%. Day to day instabilities produce a total overall variation in beam symmetry on the order of +/- 2.0%. Results suggest the measurement of symmetry as a function of gantry position become a routine quality assurance procedure for this accelerator.