Genetic evidence for a product of the Fv-1 locus that transfers resistance to mouse leukemia viruses.

Extracts of mouse cells have been shown to transfer to N- or B-trophic host range types of mouse leukemia viruses. The genetic specificity of the inhibition was tested in two ways: (i) by correlating the Fv-1 genotype of a number of mouse strains with the restriction-transferring activity of extracts of the respective embryo cell cultures, and (ii) by correlating the Fv-1 genotype of BLC3F2 (C57BL/6 female [Fv-1bb] by C3H male [Fv-1nn] parental strains) mouse embryos, which segregate the Fv-1 alleles in a 12:1 ratio, with the inhibitor activity of extracts of the cells from each embryo. Five independent matings, totaling 45 individual embryos, were tested. Each embryo was cultured, and the Fv-1 genotype was determined independently by titration of N- and B-tropic viruses; the extracts of replicate secondary cultures were tested for their effect on infection of permissive cells by N- and B-tropic viruses. The specific-restriction-transferring activity of the embryos was found to segregate with the appropriate Fv-1 genotype. These res-lts confirm the suggestion that the inhibitor of the leukemia virus host range types in the cellular extracts is a product of the Fv-1 locus.     

The Validity of a New Low-Dose Stereoradiography System to Perform 2D and 3D Knee Prosthetic Alignment Measurements

Introduction

The EOS stereoradiography system has shown to provide reliable varus/valgus (VV) measurements of the lower limb in 2D (VV2D) and 3D (VV3D) after total knee arthroplasty (TKA). Validity of these measurements has not been investigated yet, therefore the purpose of this study was to determine validity of EOS VV2D and VV3D.

Methods

EOS images were made of a lower limb phantom containing a knee prosthesis, while varying VV angle from 15° varus to 15° valgus and flexion angle from 0° to 20°, and changing rotation from 20° internal to 20° external rotation. Differences between the actual VV position of the lower limb phantom and its position as measured on EOS 2D and 3D images were investigated.

Results

Rotation, flexion or VV angle alone had no major impact on VV2D or VV3D. Combination of VV angle and rotation with full extension did not show major differences in VV2D measurements either. Combination of flexion and rotation with a neutral VV angle showed variation of up to 7.4° for VV2D; maximum variation for VV3D was only 1.5°. A combination of the three variables showed an even greater distortion of VV2D, while VV3D stayed relatively constant. Maximum measurement difference between preset VV angle and VV2D was 9.8°, while the difference with VV3D was only 1.9°. The largest differences between the preset VV angle and VV2D were found when installing the leg in extreme angles, for example 15° valgus, 20° flexion and 20° internal rotation.

Conclusions

After TKA, EOS VV3D were more valid than VV2D, indicating that 3D measurements compensate for malpositioning during acquisition. Caution is warranted when measuring VV angle on a conventional radiograph of a knee with a flexion contracture, varus or valgus angle and/or rotation of the knee joint during acquisition.     

Differential effects of interleukin-1 on hyaluronan and proteoglycan metabolism in two compartments of the matrix formed by articular chondrocytes maintained in alginate

Phenotypically stable young adult bovine articular chondrocytes suspended in beads of alginate gel were first cultured for 5 days, using daily changes of medium containing 10% fetal bovine serum and supplements. The cells in the beads were then maintained in culture for a further 3 days in the presence or absence of interleukin-1alpha at 1 ng/ml in the daily change of medium. The exposure to interleukin-1alpha caused the incorporation of (35)S-sulfate into the predominant cartilage proteoglycan, aggrecan, to decrease by approximately 60%. In addition, proteoglycans that had accumulated into the cell-associated matrix during the first 5 days of culture in the absence of interleukin-1alpha moved into the matrix further removed from the cells and from there into the medium. In contrast, the exposure to interleukin-1alpha was found to markedly promote the rate of synthesis of hyaluronan, especially during the first 24 h. Over the 3 days of culture in the presence of interleukin-1alpha, a large proportion of the newly synthesized hyaluronan molecules, as well as those that had previously become residents of the cell-associated matrix, moved out of this compartment and appeared to become permanent residents of the further removed matrix. These results demonstrate that exposure of young adult articular chondrocytes to interleukin-1alpha has profound effects on the metabolism of hyaluronan, a molecule that plays a critical role in the retention of proteoglycan molecules in the matrix. Importantly, the results suggest that exposure of chondrocytes to interleukin-1 in inflamed joints, such as occurs in rheumatoid arthritis, leads to the rapid loss of coordination of the synthesis of aggrecan and hyaluronan, two of the critical constituents of the proteoglycan aggregate. In addition, we present evidence that these interleukin-1-induced effects differentially alter the metabolism of hyaluronan in the metabolically active cell-associated matrix and the metabolically inactive matrix further removed from the chondrocytes.     

Direct stringency comparison of two macaque models (single-high vs. repeat-low) for mucosal HIV transmission using an identical anti-HIV chemoprophylaxis intervention

BACKGROUND: In our previous work, oral chemoprophylaxis with tenofovir disoproxil fumarate (TDF) provided partial protection in rhesus macaques against repeated low-dose (RL) intrarectal SHIV162p3 exposure. METHODS: Here, we make a direct comparison of these previous findings with data generated using a single high (SH)-dose challenge strategy. RESULTS: All 5 (100%) control macaques were infected after a SH challenge and only three of five (60%) TDF-treated macaques became infected. The remaining two TDF-treated macaques remained virus-negative and were susceptible to virus infection upon re-challenge in the absence of oral TDF. Thus, two of five (40%) TDF-treated macaques were protected by the pre-exposure chemoprophylaxis regimen. By comparison with the RL challenge system, only one of four (25%) of TDF-treated macaques were protected from infection, whereas four of four (100%) control macaques became infected using RL challenges. CONCLUSION: Taken together, these findings indicate that the stringency of the RL challenge model for testing antiretroviral interventions is not lower and possibly greater than that of the SH challenge model.     

Proteolytic cleavage of inducible nitric oxide synthase (iNOS) by calpain I.

Proteolytic degradation of inducible nitric oxide synthase (iNOS or NOS2; EC 1.14.13.39) is one of the key steps by which the synthetic glucocorticoid dexamethasone controls the amount of iNOS protein and thus the production of nitric oxide (NO) in interferon-gamma-stimulated RAW 264.7 cells. In the present study we examined the role of the calmodulin (CaM)-binding site present within iNOS protein for the proteolytic degradation by the calcium-dependent neutral cysteine protease calpain I (EC 3.4.22.17). Using pulse chase experiments as well as cell-free degradation assays we show that the iNOS monomer is a direct substrate for cleavage by calpain I. Two structural determinants are involved in proteolytic cleavage, the canonical CaM-binding domain present at amino acids 501-532 and a conformational determinant located within iNOS. The access of the CaM-binding region appears to be critical for substrate cleavage as incubation of in vitro synthesized iNOS with purified CaM inhibits iNOS degradation by calpain I. Moreover, cytosolic CaM levels are decreased upon treatment of RAW 264.7 cells with dexamethasone as assessed by immunoprecipitation. The data shown herein provide novel insights into the underlying mechanisms involved in the anti-inflammatory actions of glucocorticoids.     

牛脾转移因子的提取工艺研究

目的：比较从牛脾中提取活性ＴＦ的不同方法。方法：分别利用Ｌａｗｒｅｎｃｅ－－透析法、Ｌａｗｒｅｎｃｅ－超滤法、超离心－超滤法提取,再以蛋白反应、红外及紫外扫描予以鉴定比较。结果：Ｌａｗｒｅｎｃｅ－超滤法和超离心－超滤法效果较好。结论：工业化生产中应采用Ｌａｗｒｅｎｃｅ－超滤法。     

Balancing on a narrow ridge: biomechanics and control.

The balance of standing humans is usually explained by the inverted pendulum model. The subject invokes a horizontal ground-reaction force in this model and controls it by changing the location of the centre of pressure under the foot or feet. In experiments I showed that humans are able to stand on a ridge of only a few millimetres wide on one foot for a few minutes. In the present paper I investigate whether the inverted pendulum model is able to explain this achievement. I found that the centre of mass of the subjects sways beyond the surface of support, rendering the inverted pendulum model inadequate. Using inverse simulations of the dynamics of the human body, I found that hip-joint moments of the stance leg are used to vary the horizontal component of the ground-reaction force. This force brings the centre of mass back over the surface of support. The subjects generate moments of force at the hip-joint of the swing leg, at the shoulder-joints and at the neck. These moments work in conjunction with a hip strategy of the stance leg to limit the angular acceleration of the head-arms-trunk complex. The synchrony of the variation in moments suggests that subjects use a motor programme rather than long latency reflexes.