Depletion of three combined THOC5 mRNA export protein target genes synergistically induces human hepatocellular carcinoma cell death.

Hepatocellular carcinoma (HCC) is a frequent form of cancer with a poor prognosis and with limited possibilities of medical intervention. It has been shown that over 100 putative driver genes are associated with multiple recurrently altered pathways in HCC, suggesting that multiple pathways will need to be inhibited for any therapeutic method. mRNA processing is regulated by a complex RNA-protein network that is essential for the maintenance of homeostasis. THOC5, a member of mRNA export complex, has a role in less than 1% of mRNA processing, and is required for cell growth and differentiation, but not for cell survival in normal fibroblasts, hepatocytes and macrophages. In this report, we show that 50% depletion of THOC5 in human HCC cell lines Huh7 and HepG2 induced apoptosis. Transcriptome analysis using THOC5-depleted cells revealed that 396 genes, such as transmembrane BAX inhibitor motif containing 4 (TMBIM4), transmembrane emp24-like trafficking protein 10 (Tmed10) and D-tyrosyl-tRNA deacylase 2 (Dtd2) genes were downregulated in both cell lines. The depletion of one of these THOC5 target genes in Huh7 or HepG2 did not significantly induce cell death, suggesting that these may be fine tuners for HCC cell survival. However, the depletion of a combination of these genes synergistically increased the number of TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling)-positive HCC. It must be noted that the depletion of these genes did not induce cell death in the hepatocyte cell line, THLE-2 cells. THOC5 expression was enhanced in 78% of cytological differentiation grading G2 and G3 tumor in primary HCC. Furthermore, the expression of a putative glycoprotein, Tmed10, is correlated to THOC5 expression level in primary HCCs, suggesting that this protein may be a novel biomarker for HCC. These data imply that the suppression of the multiple THOC5 target genes may represent a novel strategy for HCC therapy.

Constitutive expression of murine c-FLIPR causes autoimmunity in aged mice.

Death receptor-mediated apoptosis is a key mechanism for the control of immune responses and dysregulation of this pathway may lead to autoimmunity. Cellular FLICE-inhibitory proteins (c-FLIPs) are known as inhibitors of death receptor-mediated apoptosis. The only short murine c-FLIP splice variant is c-FLIPRaji (c-FLIPR). To investigate the functional role of c-FLIPR in the immune system, we used the vavFLIPR mouse model constitutively expressing murine c-FLIPR in all hematopoietic compartments. Lymphocytes from these mice are protected against CD95-mediated apoptosis and activation-induced cell death. Young vavFLIPR mice display normal lymphocyte compartments, but the lymphocyte populations alter with age. We identified reduced levels of T cells and slightly higher levels of B cells in 1-year-old vavFLIPR mice compared with wild-type (WT) littermates. Moreover, both B and T cells from aged vavFLIPR animals show activated phenotypes. Sera from 1-year-old WT and transgenic animals were analysed for anti-nuclear antibodies. Notably, elevated titres of these autoantibodies were detected in vavFLIPR sera. Furthermore, tissue damage in kidneys and lungs from aged vavFLIPR animals was observed, indicating that vavFLIPR mice develop a systemic lupus erythematosus-like phenotype with age. Taken together, these data suggest that c-FLIPR is an important modulator of apoptosis and enforced expression leads to autoimmunity.

The role of c-FLIP splice variants in urothelial tumours.

Deregulation of apoptosis is common in cancer and is often caused by overexpression of anti-apoptotic proteins in tumour cells. One important regulator of apoptosis is the cellular FLICE-inhibitory protein (c-FLIP), which is overexpressed, for example, in melanoma and Hodgkin's lymphoma cells. Here, we addressed the question whether deregulated c-FLIP expression in urothelial carcinoma impinges on the ability of death ligands to induce apoptosis. In particular, we investigated the role of the c-FLIP splice variants c-FLIP(long) (c-FLIP(L)) and c-FLIP(short) (c-FLIP(S)), which can have opposing functions. We observed diminished expression of the c-FLIP(L) isoform in urothelial carcinoma tissues as well as in established carcinoma cell lines compared with normal urothelial tissues and cells, whereas c-FLIP(S) was unchanged. Overexpression and RNA interference studies in urothelial cell lines nevertheless demonstrated that c-FLIP remained a crucial factor conferring resistance towards induction of apoptosis by death ligands CD95L and TRAIL. Isoform-specific RNA interference showed c-FLIP(L) to be of particular importance. Thus, urothelial carcinoma cells appear to fine-tune c-FLIP expression to a level sufficient for protection against activation of apoptosis by the extrinsic pathway. Therefore, targeting c-FLIP, and especially the c-FLIP(L) isoform, may facilitate apoptosis-based therapies of bladder cancer in otherwise resistant tumours.

Investigations of ascorbic acid interference in urine test strips.

Ascorbic acid at higher concentration in urine samples can lead to false negative results in a number of urine tests, with a potential risk of clinical findings being overlooked, particularly with glucose and hemoglobin. For this reason, the ascorbic acid status of urine samples should always be routinely known so as to establish what adjustment needs to be made. A much better approach, however, is to use a test which is by design largely resistant to ascorbic acid. We compared five very common 10-parameter urine test strips from different manufacturers. The results of this study show that of the strips tested, only the product Combur-Test from Roche Diagnostics is largely resistant to ascorbic acid interference. Even lowest - but clinically relevant - concentrations of erythrocytes (10/microL), hemoglobin (0.03 mg/dL), and glucose (50 mg/dL) were correctly detected with concentrations of up to 400 mg/L ascorbic acid. Higher analyte concentrations correctly reacted positive even in the presence of up to 1000 mg/L ascorbic acid.

Observation of laser-pulse shortening in nonlinear plasma waves.

We have measured the temporal shortening of an ultraintense laser pulse interacting with an underdense plasma. When interacting with strongly nonlinear plasma waves, the laser pulse is shortened from 38 +/- 2 fs to the 10-14 fs level, with a 20% energy efficiency. The laser ponderomotive force excites a wakefield, which, along with relativistic self-phase modulation, broadens the laser spectrum and subsequently compresses the pulse. This mechanism is confirmed by 3D particle in cell simulations.

Spatial characteristics of Kalpha x-ray emission from relativistic femtosecond laser plasmas.

The spatial structure of the Kalpha emission from Ti targets irradiated with a high intensity femtosecond laser has been studied using a two-dimensional monochromatic imaging technique. For laser intensities I<5 x 10(17) W/cm(2), the observed spatial structure of the Kalpha emission can be explained by the scattering of the hot electrons inside the solid with the help of a hybrid particle-in-cell/Monte Carlo model. By contrast, at the maximum laser intensity I=7 x 10(18) W/cm(2) the half-width of the Kalpha emission was 70 microm compared to a laser-focus half-width of 3 microm. Moreover, the main Kalpha peak was surrounded by a halo of weak Kalpha emission with a diameter of 400 microm and the Kalpha intensity at the source center did not increase with increasing laser intensity. These three features point to the existence of strong self-induced fields, which redirect the hot electrons over the target surface.

High-intensity laser induced ion acceleration from heavy-water droplets.

Fusion neutrons from a heavy water droplet target irradiated with laser pulses of 3 x 10(19) W/cm(2) and from a deuterated secondary target are observed by a time-of-flight (TOF) neutron spectrometer. The observed TOF spectrum can be explained by fusion of deuterium ions simultaneously originating from two different sources: ion acceleration in the laser focus by ponderomotively induced charge separation and target-normal sheath acceleration off the target rear surface. The experimental findings agree well with 3D particle-in-cell simulations.

The use of pulmonary function testing in piloting, air travel, mountain climbing, and diving.

Millions of people engage in occupational or leisure activities at high altitude or at variable depths below sea level. This article presents an overview of the utility of pulmonary function testing in evaluating complications and other consequences of exposure to high and low pressure environments. The authors review recent literature concerning expected changes in pulmonary function with hyperbaric and hypobaric exposures. The article provides guidance for clinicians evaluating mountain climbers, pilots, aircrew members, airline passengers and deep sea divers.

The precautionary principle: between responsibility and policy. / Le principe de précaution entre responsabilité et politique.

It is important to distinguish between the idea of the "precautionary principle", a precise concept that made its appearance under specific circumstances in the 70s in order to respond to certain environment-related issues, and the idea of "precaution", itself more or less confused with the idea of "caution", a term used to describe a general attitude to risk and uncertainty. Although it is easy to identify the notion of "precaution" with a general philosophy of responsibility, the "precautionary principle" has no meaning except when used to describe public precautionary policies. And though we cannot give a precise definition of the idea of precaution, in the sense of caution, we can however give a precise description of the "precautionary principle", based on the techniques, always specific, which are implemented in the name of the precautionary principle. In France we have developed the habit of considering precautionary actions as part of an overall moral principle, while totally disregarding the laws defining the "precautionary principle" and including it within the power framework of state policy. It is essential to clear up this confusion, along with all of its potentially dangerous legal consequences, before we can envisage an informed discussion on the "precautionary principle", its field of application and the conditions governing its implementation.

Kinematic total knee arthroplasty: a 10- to 14-year prospective follow-up review.

To understand better the type and incidence of long-term complications in total knee replacement, 306 primary Kinematic total knee arthroplasties performed between June 1978 and December 1982 were prospectively reviewed in detail. The Kinematic knee is a nonconstrained, posterior cruciate-retaining prosthesis that has right and left femoral components to afford anatomic tracking of the patella. The overall revision rate was 6.5%. The most common cause for revision surgery was patellar complications. Ten revisions (3.06%) were for patellar component loosening. Two knees were revised for patella subluxation (0.65%); 1 was in a resurfaced rheumatoid patella, and 1 in an unsurfaced osteoarthritic patella. Stair climbing was better with an unsurfaced patella. Anterior knee pain was 21.8% in the unsurfaced patella and 11.2% in the replaced patella. These data suggest patella replacement is not appropriate with this design.

Results of total elbow arthroplasty after excision of the radial head and synovectomy in patients who had rheumatoid arthritis.

We compared the results of twenty-three consecutive capitellocondylar total elbow arthroplasties in twenty-three patients in whom an excision of the radial head and synovectomy for rheumatoid arthritis had failed with those of twenty-three non-consecutive primary capitellocondylar total elbow arthroplasties in twenty-three patients who had rheumatoid arthritis. The two groups were matched for age, gender, duration of follow-up, side of the operation, type of prosthesis, and operative approach. The average duration of follow-up was four years (range, two to fourteen years). At the most recent follow-up examination, use of a 100-point rating system demonstrated an improvement from an average preoperative score of 21 points (range, 12 to 42 points) to an average postoperative score of 87 points (range, 17 to 97 points) for the group in whom an excision of the radial head and synovectomy had failed. The group that had primary arthroplasty demonstrated an improvement from an average preoperative score of 22 points (range, 7 to 42 points) to an average postoperative score of 94 points (range, 85 to 100 points). The group that had primary arthroplasty had a significantly greater improvement in terms of relief of pain (p < 0.05), functional status (p < 0.01), and the elbow-rating score (p < 0.03) than the other group. Four patients who had had failure of an excision of the radial head and synovectomy and none of those who had primary arthroplasty needed an additional operative procedure. Six of the patients who had had a failed excision and synovectomy and none of the patients who had primary arthroplasty had instability of the elbow components. We concluded that, although excision of the radial head and synovectomy is a conservative and effective method of treating a painful rheumatoid elbow, conversion to a capitellocondylar total elbow arthroplasty is more difficult after such an operation and the results at a minimum of two years are inferior to those for primary capitellocondylar total elbow arthroplasty.

Total knee arthroplasty in a group of patients less than 45 years of age.

The long-term follow-up evaluation of total knee arthroplasty (TKA) in patients under age 45 is reviewed. One hundred three knees in 67 patients who had an average follow-up period of 7.2 years were retrospectively reviewed. Fifty-eight percent of the patients had rheumatoid arthritis, and 29% had juvenile rheumatoid arthritis. Thirteen percent of the patients had post-traumatic arthritis, avascular necrosis, hemochromatosis, or lupus. The results demonstrate that the success of TKA in this patient population are comparable to those for TKA in the elderly.

Analysis of the inspiratory flow-volume curve. Should it always precede the forced expiratory maneuver?

The inspiratory flow-volume (FV) curve can be used to identify patients with upper airway obstruction, air trapping, and restriction. Current computed pulmonary function testing equipment often mandates a forced expiratory maneuver (FEM) immediately prior to the forced inspiratory maneuver (standard method). We evaluated the inspiratory FV curve with and without an antecedent FEM in 119 subjects referred for pulmonary function testing. The subjects were divided into four groups by grading the degree of airway obstruction using confidence intervals of the FEV1/FVC percent predicted minus the actual FEV1/FVC percent measured from the best FEM according to Intermountain Thoracic Society recommendations. The forced inspiratory vital capacity (FIVC), forced inspiratory flow 50 (FIF50), and peak inspiratory flow (PIF) from the inspiratory FV curve with an antecedent FEM was compared with the FIVC, FIF50, and PIF without an antecedent FEM in each category of obstructive lung disease. The FIVC without the antecedent FEM was significantly larger than that with an antecedent FEM by 170 ml (p < 0.002) in subjects with severe airway obstruction, but was not significantly different in the other groups. The FIF50 was not significantly different in any group, but approached significance in both normal subjects and subjects with severe obstruction. The PIF was not significantly different in any group, but approached significance in the normal subjects, order for patients with severe obstructive airway disease to generate a valid forced inspiratory FV curve, it should be obtained without an antecedent FEM. When a plateau of the inspiratory FV curve is encountered, we suggest that is useful to generate the inspiratory FV curve prior to the FEM and to analyze its flow and volume characteristics independent of the FEM. The "best" inspiratory FV curve should then be displayed with the "best" FEM for proper evaluation of the FV loop.

Distal loss of femoral bone following total knee arthroplasty. Measurement with visual and computer-processing of roentgenograms and dual-energy x-ray absorptiometry.

The changes in bone-mineral content in the distal aspect of a cadaveric femur that had been prepared for insertion of the femoral component of a total condylar knee prosthesis were evaluated with visual inspection and computer-processing of roentgenograms and with dual-energy x-ray absorptiometry. Seventeen small slices of bone, each three millimeters wide, were removed so that, finally, 89 per cent of the bone was lost from the distal femoral metaphysis. Standardized lateral roentgenograms of the specimen were made with use of a reference step-wedge of hydroxyapatite, and dual-energy x-ray absorptiometry studies were performed with the x-ray beam tangential to the interface. The roentgenograms were digitized and the bone mineral was measured with use of computer analysis. Dual-energy x-ray absorptiometry was performed with and without the femoral prosthesis in place, in order to determine the effect of the metallic prosthesis on the accuracy of the measurement. A bone loss of 25 per cent or more was identified visually by all five of the readers 100 per cent of the time; losses of 20 to 24 per cent, 15 to 19 per cent, 10 to 14 per cent, and 3 to 9 per cent were correctly identified 92, 75, 66, and 59 per cent of the time, respectively. The measurements of bone-mineral content that were obtained from the digitized roentgenograms were linearly correlated with the actual bone-mineral content (the ash content) (r = 0.97, p < 0.001) and were three times more accurate than the visual readings. The determinations of bone-mineral content with dual-energy x-ray absorptiometry correlated highly with the ash content (r = 1.00, p < 0.001) and were seven times more accurate than the visual readings. There was only a 4 per cent difference between the measurements with dual-energy x-ray absorptiometry made with the prosthesis in place and those made without it in place (p < 0.01). Dual-energy x-ray absorptiometry was the most accurate of the three methods and could detect the smallest experimentally created loss; computer-processing and visual-processing of roentgenograms detected losses of 8 per cent or more and 25 per cent or more, respectively. Dual-energy x-ray absorptiometry and computer-processing of the roentgenograms quantified the bone loss, while visual-processing could detect only the presence or absence of bone loss.(ABSTRACT TRUNCATED AT 400 WORDS)

Long-term complications after total knee arthroplasty with or without resurfacing of the patella.

The long-term complications related to the patella were retrospectively evaluated for 891 knees (684 patients) that had had a total arthroplasty, with or without resurfacing of the patella, with use of an unconstrained, condylar, posterior-cruciate-preserving prosthesis. The study population comprised two groups of patients who were similar in size, age, sex distribution, and diagnosis. One group (396 knees [303 patients]) had had a total knee arthroplasty with patellar resurfacing and the other group (495 knees [381 patients]) had had the same procedure without resurfacing. The average duration of follow-up was six and one-half years (range, two to fifteen years). The decision to resurface the patella was based on subjective inspection of the articular surface and on assessment of patellar tracking at the time of the operation. Resurfacing was performed if there was loss of cartilage, exposed bone, gross surface irregularities, or tracking abnormalities. Complications occurred an average of three years (range, immediately postoperatively to nine years) after the operation in the group that had had resurfacing and an average of four years (range, immediately post-operatively to ten years) postoperatively in the group that had not had resurfacing. In the group that had had resurfacing, there was loosening of the patellar component in five knees, patellar subluxation in four knees, fracture of the patella in three knees, rupture of the patellar tendon in three knees, and chronic peripatellar pain in one knee. In the group that had not had resurfacing, the complications included patellar subluxation in five knees, rupture of the patellar tendon in two knees, and chronic peripatellar pain in fifty-one knees.(ABSTRACT TRUNCATED AT 250 WORDS)

Capitellocondylar total elbow replacement in rheumatoid arthritis. Long-term results.

We evaluated the long-term results of 202 capitellocondylar total elbow replacements that had been performed, from July 1974 through June 1987, in 172 patients. The duration of follow-up averaged sixty-nine months (range, twenty-four to 178 months). At the most recent follow-up examination, use of a 100-point rating score demonstrated an improvement from an average preoperative score of 26 points (range, 2 to 50 points) to an average postoperative score of 91 points (range, 45 to 100 points). The most improvement occurred in the categories of relief of pain, functional status, and range of motion in all planes except extension. The improvements in these categories and in the roentgenographic appearance that were seen in the early postoperative period did not deteriorate with time. The average preoperative arc of motion at the elbow ranged from -37 degrees of extension to 118 degrees of flexion. The average postoperative arc of motion at the elbow ranged from -30 degrees of extension to 135 degrees of flexion. Supination improved from 45 degrees preoperatively to 64 degrees postoperatively; pronation improved from 56 degrees preoperatively to 72 degrees postoperatively. The roentgenograms showed a radiolucent line adjacent to eight humeral and nineteen ulnar components; most of the lines were incomplete and one millimeter wide or less. Revision of the prosthesis was necessary in three elbows (1.5 per cent) because of loosening without infection, and in three additional elbows because of dislocation of the prosthesis. Complications included deep infection in three elbows (1.5 per cent); problems related to the wound in fifteen (7 per cent); permanent, partial sensory ulnar-nerve palsy in five (2.5 per cent); permanent, partial motor ulnar-nerve palsy in one (0.5 per cent); and dislocation in seven (3.5 per cent).