Assessment of quadriceps muscle performance by hand-held, isometric, and isokinetic dynamometry in patients with knee dysfunction.

One component of patient evaluation is muscle performance assessment. The purpose of this study was: 1) to determine the difference and correlation between hand-held, isometric, and isokinetic dynamometry test results in patients with knee dysfunction and 2) to determine the effect of pain during such testing. Bilateral quadriceps strength in 23 subjects with unilateral knee dysfunction was tested using concentric and eccentric isokinetic dynamometry at 60 degrees/sec, isometric dynamometry, and hand-held dynamometry, both at 60 degrees of knee flexion. Pain ratings were obtained before, during, and after each test. Statistical analysis revealed a significant difference between involved and uninvolved limbs for eccentric isokinetic dynamometry (p = 0.002) and hand-held dynamometry (p = 0.005); no difference was found between limbs for the concentric isokinetic and isometric dynamometry (p > 0.05). Mean percent deficits in quadriceps strength ranged from 11 to 18%, with no significant difference found between testing modes. Pearson product moment correlations ranged from 0.34 to 0.76 when comparing testing modes. No significant difference existed in pain scores before, during, and after each mode of testing. It was concluded that large variation existed between different testing modes, which results in different conclusions regarding the strength of the quadriceps in patients with knee dysfunction.

Effect of patellar taping and bracing on patellar position as determined by MRI in patients with patellofemoral pain.

OBJECTIVE: To determine the effects of patellar taping, bracing, and not taping on patellar position. DESIGN AND SETTING: An experimental design was used to compare patellar taping, bracing, and not taping on patellar position as determined by magnetic resonance imaging (MRI). SUBJECTS: Twelve subjects with a diagnosis of patellofemoral pain participated in this study. MEASUREMENTS: Static MRI images were taken at 8 angles of knee flexion (10, 16, 25, 30, 34, 39, 41, and 45 degrees ). Patellofemoral congruence angle (PFC), lateral patellar displacement (LPD), and lateral patellar angle (LPA) were determined by digitization. RESULTS: A repeated-measures multivariate analysis of variance was used to compare experimental conditions. Across all angles of knee flexion, a more lateral PFC existed for the control condition (-4.1 degrees ) than the brace condition (-7.1 degrees ) or tape condition (-6.1 degrees ). Post hoc testing revealed that this difference was statistically different only at 10 degrees of knee flexion. Across all knee angles, LPD was more medial for the braced condition (1.7 mm) than for the tape (2.7 mm) or control (2.6 mm) condition. Post hoc testing revealed that this difference was statistically different only at 10 degrees of knee flexion. No differences existed between conditions for LPA. CONCLUSIONS: We conclude that patellar bracing and taping influenced patellar position (PFC and LPD) at 10 degrees of knee flexion during a static MRI condition.

gag, vif, and nef genes contribute to the homologous viral interference induced by a nonproducer human immunodeficiency virus type 1 (HIV-1) variant: identification of novel HIV-1-inhibiting viral protein mutants.

We previously demonstrated that expression of the nonproducer F12-human immunodeficiency virus type 1 (HIV-1) variant induces a block in the replication of superinfecting HIV that does not depend on the down-regulation of CD4 HIV receptors. In order to individuate the gene(s) involved in F12-HIV-induced interference, vectors expressing each of the nine F12-HIV proteins were transfected in HIV-susceptible HeLa CD4 cells. Pools of cell clones stably producing each viral protein were infected with HIV-1, and virus release was measured in terms of reverse transcriptase activity in supernatants. We hereby demonstrate that HeLa CD4 cells expressing the F12-HIV gag, vif, or nef gene were resistant, to different degrees, to infection with T-cell-line-adapted HIV-1 strains. Conversely, expression of either the tat, rev, or vpu F12-HIV gene increased the rate of HIV release, and no apparent effects on HIV replication were observed in cells expressing either the F12-HIV vpr, pol, or env gene. No variation of CD4 exposure was detected in any of the uninfected HeLa CD4 pools. These data indicate that F12-HIV homologous viral interference is the consequence of the synergistic anti-HIV effects of Gag, Vif, and Nef proteins. Retrovirus vectors expressing F12-HIV vif or nef allowed us to further establish that the expression of each mutated protein (i) inhibits the replication of clinical HIV-1 isolates as well, (ii) impairs the infectivity of the virus released by cells chronically infected with HIV-1, and (iii) limitedly to F12-HIV Vif protein, induces HIV resistance in both vif-permissive and vif-nonpermissive cells. The levels of action of F12-HIV vif and nef anti-HIV effects were also determined. We observed that HIV virions emerging from the first viral cycle on F12-HIV vif-expressing cells, although released in unaltered amounts, had a strongly reduced ability to initiate the retrotranscription process when they reinfected parental HeLa CD4 cells. Differently, we observed that expression of F12-HIV Nef protein affects the HIV life cycle at the level of viral assembling and/or release. For the first time, an inhibitory effect on the HIV life cycle in both acutely and chronically infected cells induced by mutated Vif and Nef HIV-1 proteins is described. These genes could thus be proposed as new useful reagents for anti-HIV gene therapy.

Anti-HIV viral interference induced by retroviral vectors expressing a nonproducer HIV-1 variant.

A Hut-78 cell clone (F12) harboring a nonproducer human immunodeficiency virus (HIV-1) variant shows complete resistance to HIV-1 or HIV-2 superinfection. The F12-HIV provirus produces an altered HIV-1 protein pattern and cannot generate even immature viral particles. We demonstrated that HeLa CD4+ cells transfected with the F12-HIV genome resist HIV superinfection through a CD4-independent mechanism. As F12-HIV appears to be a useful system to induce anti-HIV intracellular immunization, we constructed various retroviral vectors containing the F12-HIV genome, modified by elimination of the F12 3'LTR and part of its nef gene, inserted 'antisense' with respect to the Moloney murine leukemia virus 5' LTR. Here we show that recombinant retroviral particles carrying the N2/F12-HIV nef- (as) construct can stably transduce both CEMss human cells and primary human peripheral blood lymphocytes, inducing the expression of the F12-HIV genome. These results could open the way to an anti-AIDS gene therapy strategy based on F12-HIV-induced intracellular immunization.

cis expression of the F12 human immunodeficiency virus (HIV) Nef allele transforms the highly productive NL4-3 HIV type 1 to a replication-defective strain: involvement of both Env gp41 and CD4 intracytoplasmic tails.

F12 human immunodeficiency virus type 1 (HIV-1) nef is a naturally occurring nef mutant cloned from the provirus of a nonproductive, nondefective, and interfering HIV-1 variant (F12-HIV). We have already shown that cells stably transfected with a vector expressing the F12-HIV nef allele do not downregulate CD4 receptors and, more peculiarly, become resistant to the replication of wild type (wt) HIV. In order to investigate the mechanism of action of such an HIV inhibition, the F12-HIV nef gene was expressed in the context of the NL4-3 HIV-1 infectious molecular clone by replacing the wt nef gene (NL4-3/chi). Through this experimental approach we established the following. First, NL4-3/chi and nef-defective (Deltanef) NL4-3 viral particles behave very similarly in terms of viral entry and HIV protein production during the first replicative cycle. Second, no viral particles were produced from cells infected with NL4-3/chi virions, whatever the multiplicity of infection used. The viral inhibition apparently occurs at level of viral assembling and/or release. Third, this block could not be relieved by in-trans expression of wt nef. Finally, NL4-3/chi reverts to a producer HIV strain when F12-HIV Nef is deprived of its myristoyl residue. Through a CD4 downregulation competition assay, we demonstrated that F12-HIV Nef protein potently inhibits the CD4 downregulation induced by wt Nef. Moreover, we observed a redistribution of CD4 receptors at the cell margin induced by F12-HIV Nef. These observations strongly suggest that F12-HIV Nef maintains the ability to interact with the intracytoplasmic tail of the CD4 receptor molecule. Remarkably, we distinguished the intracytoplasmic tails of Env gp41 and CD4 as, respectively, viral and cellular targets of the F12-HIV Nef-induced viral retention. For the first time, the inhibition of the viral life cycle by means of in-cis expression of a Nef mutant is here reported. Delineation of the F12-HIV Nef mechanism of action may offer additional approaches to interference with the propagation of HIV infection.