Effect of fatigue load on internal mechanical properties of the intervertebral disc / Efecto de la carga de fatiga en las propiedades mecánicas internas del disco intervertebral

SUMMARY: Lumbar disc herniation is considered to be the main pathological factor for the common clinical disease of low back pain. Biomechanical factor is an important cause of lumbar disc herniation, so it is urgent to analyze the stress/strain behavior of intervertebral disc under different loading condition. Slow repetitive loading is considered to be an important factor of spine and disc injuries, and the effect of fatigue load on internal displacement in the intervertebral disc was investigated by applying the optimized digital image correlation technique in this study. The first finding was that fatigue load had a significant effect on the displacement distribution in the intervertebral disc under compression. Superficial AF exhibited the largest axial displacements before fatigue load, while it exhibited the smallest axial displacements after fatigue load. Inner AF exhibited slightly smaller radial displacements than outer AF before fatigue load, while it exhibited significantly greater radial displacements than outer AF displacements after fatigue load. The second finding was that fatigue load had a certain effect on the internal displacement distribution in the flexed intervertebral disc under compression. Middle AF exhibited the smallest axial displacements before fatigue load, while deep AF exhibited the smallest axial displacements after fatigue load. The radial displacement distribution did not change before and after fatigue load, as the radial displacement in outer AF was the smallest, while the radial displacement in inner AF was the largest. The third finding was that with the increase in fatigue time and amplitude, the Young's modulus of the intervertebral disc increased significantly. This study can provide the basis for clinical intervertebral disc disease prevention and treatment? and is important for mechanical function evaluation of artificial intervertebral disc as well.

RESUMEN: La hernia de disco lumbar se considera el principal factor patológico para la enfermedad clínica común del dolor lumbar. El factor biomecánico es una causa importante de hernia de disco lumbar, por lo que es urgente analizar el comportamiento de esfuerzo / tensión del disco intervertebral bajo diferentes condiciones de carga. La carga repetitiva lenta se considera un factor importante de lesiones de columna y disco, y en este estudio el efecto de la carga de fatiga sobre el desplazamiento interno en el disco intervertebral se investigó mediante la aplicación de la técnica de correlación de imagen digital optimizada. El primer hallazgo fue que la carga de fatiga tuvo un efecto significativo en la distribución del desplazamiento en el disco intervertebral bajo compresión. El AF superficial exhibió los desplazamientos axiales más grandes antes de la carga de fatiga, mientras que exhibió los desplazamientos axiales más pequeños después de la carga de fatiga. El AF interno exhibió desplazamientos radiales ligeramente más pequeños que el AF externo antes de la carga de fatiga, mientras que exhibió desplazamientos radiales significativamente mayores que los desplazamientos AF externos después de la carga de fatiga. El segundo hallazgo fue que la carga de fatiga tenía un cierto efecto sobre la distribución del desplazamiento interno en el disco intervertebral flexionado bajo compresión. El AF medio exhibió los desplazamientos axiales más pequeños antes de la carga de fatiga, mientras que el AF profundo exhibió los desplazamientos axiales más pequeños después de la carga de fatiga. La distribución del desplazamiento radial no cambió antes ni después de la carga de fatiga, ya que el desplazamiento radial en la FA externa fue el más pequeño, mientras que el desplazamiento radial en la FA interna fue el más grande. El tercer hallazgo fue que con el aumento del tiempo de fatiga y la amplitud, el módulo de Young del disco intervertebral aumentó significativamente. Este estudio puede proporcionar la base para la prevención y el tratamiento clínico de la enfermedad del disco intervertebral, y también es importante para la evaluación de la función mecánica del disco intervertebral artificial.

Expression and purification of soluble single-chain Fv against human fibroblast growth factor receptor 3 fused with Sumo tag in Escherichia coli

Background Overexpression or mutated activation of Fibroblast growth factor receptor 3 (FGFR3) is involved in the pathogenesis of many tumors. More and more studies focus on the potential usage of therapeutic antibodies against FGFR3. Results In this study, a novel single-chain Fv (ScFv) against FGFR3 was prepared and characterized. To achieve the soluble expression, ScFv was fused with Sumo (Small ubiquitin-related modifier) by polymerase chain reaction (PCR), and cloned into pET-20b. The recombinant bacteria were induced by 0.5 mM Isopropyl-ß-d-thiogalactopyranoside (IPTG) for 16 h at 20°C, and the supernatant liquid of Sumo-ScFv was harvested and purified by Ni-NTA chromatography. After being cleaved by the Sumo protease, the recombinant ScFv was released from the fusion protein, and further purified by Ni-NTA chromatography. The purity of ScFv was shown to be higher than 95% and their yield reached 4 mg per liter of bacterial culture. In vitro data showed that ScFv can significantly attenuate FGF9-induced phosphorylation of FGFR3. Conclusion We provide a novel method to produce soluble expression and bioactive functions of ScFv in Escherichia coli.

Allelic diversity at the Merozoite surface protein-1 (MSP-1) locus in natural Plasmodium falciparum populations: a brief overview

The merozoite surface protein-1 (MSP-1) locus of Plasmodium falciparum codes for a major asexual blood-stage antigen currently proposed as a major malaria vaccine candidate. The protein, however, shows extensive polymorphism, which may compromise its use in sub-unit vaccines. Here we compare the patterns of allelic diversity at the MSP-1 locus in wild isolates from three epidemiologically distinct malaria-endemic areas: the hypoendemic southwestern Brazilian Amazon (n=54), the mesoendemic southern Vietnam (n=238) and the holoendemic northern Tanzania (n=79). Fragments of the variable blocks 2, 4a, 4b and 6 or 10 of this single-copy gene were amplified by the polymerase chain reaction, and 24 MSP-1 gene types were defined as unique combinations of allelic types in each variable block. Ten different MSP-1 types were identified in Brazil, 23 in Vietnam and 13 in Tanzania. The proportion of genetically mixed infections (isolates with carrying more one MSP-1 version) ranged from 39 per cent in Brazil to 44 per cent in Vietnam and 60 per cent in Tanzania. The vast majority (90 per cent) of the typed parasite populations from Brazil and Tanzania belonged to the same seven most frequent MSP-1 gene types. In contrast, these seven types corresponded to only 61 per cent of the typed parasite populations from Vietnam. Non-random associations were found between allelic types in blocks 4a and 6 among Vietnamese isolates, the same pattern being observed in independent studies performed in 1994, 1995 and 1996. These results suggest that MSP-1 is under selective pressure in the local parasite population. Nevertheless, the finding that similar MSP-1 type frequencies were found in 1994 and 1996 argues against the prominence of short-term frequency-dependent immune selection of MSP-1 polymorphisms. Non-random associations between MSP-1 allelic types, however, were not detected among isolates from Brazil and Tanzania. A preliminary analysis of the distribution od MSP-1 gene types per host among isolates from Tanzania, but not among those from Brazil and Vietnam, shows significant deviation from that expected under the null hypothesis of independent distribution of parasites carrying different gene types in the human hosts. Some epidemiological consequences of these findings are discussed.