Tumor buding y el cáncer colorrectal / Budding tumour and colorectal cancer

Introduction: Tumour budding (TB) is defined as the presence of clusters of tumoural cells detaching from invasive margin of main tumor. It is an independent adverse prognostic factor in colorectal cancer. The aim of this study is to determinate if severity of tumor budding is associated with others prognostic factors in colorectal cancer. Materials and Methods: The study group is composed by 43 patients (27 males and 16 females; average age 73.4 years, (27-91) with colorectal cancer who underwent curative surgery. The histologic method of tumour budding used in this study was described by Nakamura. The applied statistical software was G-Stat 2.0. Statistical significance is accepted at p < 0.05. Results: High grade of TB was significantly associated with lymph node metastasis (p = 0.027), infiltrative tumour-border configuration (p = 0.016), lymphvessels invasion (p = 0.02), perineural invasion (p = 0.009) and tumor deposits (p = 0.018). There was a significant association with low grade of TB and peritumoural lymphocytic infiltration (p = 0.004). Conclusions: High grade of TB is significantly associated with other adverse prognostic factors as lymph node metastasis, infiltrative tumour-border configuration, lymphvessels invasion, venous invasion, perineural invasion and tumor deposits; and low grade of TB with favorable prognostic factor as peritumoural lymphocytic infiltration in colorectal cancer. Tumour budding can help to identify hig-risk patients with colorectal cancer.

Introducción: El tumor budding (TB) es la presencia de células tumorales aisladas o en pequeños grupos situadas en el frente infiltrante del tumor. Su hallazgo en alto grado es un factor de mal pronóstico independiente del cáncer colorrectal. El objetivo de este trabajo es determinar si el grado de TB está asociado con otros factores pronósticos del cáncer colorrectal. Materiales y Métodos: Se analizaron retrospectivamente 43 pacientes (27 varones y 16 mujeres) con una edad media de 73,4 años (27-91) intervenidos por cáncer colorrectal. El método histológico utilizado para determinar la presencia de TB fue el descrito por Nakamura en 2005. El análisis estadístico se realizó con el programa G-Stat2.0. Las diferencias se consideraron significativas si p < 0,05. Resultados: La presencia de TB de alto grado se asocia significativamente con la presencia de metástasis a ganglios linfáticos (p = 0,027), patrón de crecimiento infiltrativo (p = 0,016), invasión linfática (p = 0,02), perineural (p = 0,009) y depósitos tumorales discontinuos (p = 0,018). El TB de bajo grado se relaciona con la presencia de reacción linfocitaria peritumoral (p = 0,004). Conclusiones: El tumor budding alto grado se asocia con otros factores de mal pronóstico, como metástasis a ganglios linfáticos, crecimiento infiltrativo, invasión linfática, perineural y depósitos tu-morales discontinuos; y el tumor budding bajo grado con factores de buen pronóstico del cáncer colorrectal como reacción linfocitaria peritumoral. El análisis del grado de tumor budding podría ayudar a identificar a pacientes de peor pronóstico con cáncer colorrectal.

An approach to the in vitro evaluation of potential for cytochrome P450 enzyme inhibition from herbals and other natural remedies.

Herbals and other natural remedies could affect the disposition of conventional pharmaceuticals through inhibition of human cytochrome P-450 (CYP) enzymes. We have developed an approach to the problem of determining a critical potential for CYP enzyme inhibition by commercial herbal remedies etcetera using standardised extraction procedures in combination with commercially available human CYP enzyme 3A4, 2D6 and 2C19 inhibition assays. We present a survey of commercially available natural remedies on the local market using this approach together with a discussion on how to decide when further in vivo interaction studies may be warranted. We confirmed earlier findings on Hypericum (St. John's wort) and Echinacea purpurea activities, and report that extracts of Valeriana as well as a fish oil preparation were potent inhibitors of all tested enzymes. As a first estimate to assess the relevance of in vitro findings, we have chosen to express the inhibitory potency as the volume that the extractable inhibitory activity for a recommended human dose could be diluted into to yield a half-maximal inhibition--litres per dose unit. We propose that preparations for which this measure approaches four litres/dose unit, i.e. corresponding to the human blood volume, should be investigated further for potential enzyme interaction with pharmaceuticals.

In situ forces in the human posterior cruciate ligament in response to muscle loads: a cadaveric study.

The objectives of this study were to determine the effects of hamstrings and quadriceps muscle loads on knee kinematics and in situ forces in the posterior cruciate ligament of the knee and to evaluate how the effects of these muscle loads change with knee flexion. Nine human cadaveric knees were studied with a robotic manipulator/universal force-moment sensor testing system. The knees were subjected to an isolated hamstrings load (40 N to both the biceps and the semimembranosus), a combined hamstrings and quadriceps load (the hamstrings load and a 200-N quadriceps load), and an isolated quadriceps load of 200 N. Each load was applied with the knee at full extension and at 30, 60, 90, and 120 degrees of flexion. Without muscle loads, in situ forces in the posterior cruciate ligament were small, ranging from 6+/-5 N at 30 degrees of flexion to 15+/-3 N at 90 degrees. Under an isolated hamstrings load, the in situ force in the posterior cruciate ligament increased significantly throughout all angles of knee flexion, from 13+/-6 N at full extension to 86+/-19 N at 90 degrees. A posterior tibial translation ranging from 1.3+/-0.6 to 2.5+/-0.5 mm was also observed from full extension to 30 degrees of flexion under the hamstrings load. With a combined hamstrings and quadriceps load, tibial translation was 2.2+/-0.7 mm posteriorly at 120 degrees of flexion ut was as high as 4.6+/-1.7 mm anteriorly at 30 degrees. The in situ force in the posterior cruciate ligament decreased significantly under this loading condition compared with under an isolated hamstrings load, ranging from 6+/-7 to 58+/-13 N from 30 to 120 degrees of flexion. With an isolated quadriceps load of 200 N, the in situ forces in the posterior cruciate ligament ranged from 4+/-3 N at 60 degrees of flexion to 34+/-12 N at 120 degrees. Our findings support the notion that, compared with an isolated hamstrings load, combined hamstrings and quadriceps loads significantly reduce the in situ force in the posterior cruciate ligament. These data are in direct contrast to those for the anterior cruciate ligament. Furthermore, we have demonstrated that the effects of muscle loads depend significantly on the angle of knee flexion.

Quantitative analysis of human cruciate ligament insertions.

The objective of this study was to provide quantitative data on the insertion sites of the cruciate ligaments. In the first part of the study, we determined the shapes and sizes of the insertions of the anterior and posterior cruciate ligaments (ACL and PCL), and further compared these data with the midsubstance cross-sectional areas of the ligaments. The cross-sectional area of the ACL and PCL midsubstance of 5 human knees was measured using a laser micrometer system. The insertion sites of each ligament were then digitized and the 2-dimensional insertion site areas were determined. Relative to the ligament midsubstance, the PCL tibial and femoral insertions were approximately 3 times larger, whereas those of the ACL were over 3.5 times larger. In the second part of the study, the ACLs and PCLs of 10 knees were each divided into their 2 components and the areas of each insertion were determined. Each component was approximately 50% of the total ligament insertion area and no significant difference between the 2 could be shown.

Cytokine-induced tendinitis: a preliminary study in rabbits.

This study was designed to determine the effects of a single injection of a species-specific preparation of cytokines into rabbit patellar tendons and to compare the results with a known model of tendinitis, the collagenase-injection model. New Zealand White rabbits were divided into two groups and two time periods (4 and 16 weeks) and injected in the midsubstance of the right patellar tendon with either cytokines or collagenase under ultrasound guidance to confirm intratendinous needle placement. The left patellar tendon was injected with 0.025 ml of saline solution and served as a control. The rabbits were returned to cage activity after injection. At death, two rabbits in each group underwent histological analysis; the remaining eight animals in each time frame were evaluated biomechanically and then biochemically with use of the patella/whole patellar tendon/tibia complex. Histologic results at 4 weeks in the tendons injected with cytokines demonstrated increased cellularity, which was resolving by 16 weeks. The matrix appeared unchanged. The tendons injected with collagenase demonstrated increased angiogenesis of the matrix, hypercellularity, and fibrosis around the tendon at 4 weeks. At 16 weeks, myxoid changes, focal fibrosis, and collagen-bundle disarray with persistent increase in cellularity were noted. Biomechanically, a significant decrease in ultimate load at 16 weeks was seen in the tendons injected with cytokines but no change was seen in cross-sectional area. The tendons injected with collagenase demonstrated a significant increase in cross-sectional area at 4 and 16 weeks compared with those injected with cytokines. Biochemically, there was no significant difference in collagen content between the two groups at 4 or 16 weeks but the tendons injected with collagenase demonstrated a significant increase in crosslinking at 16 weeks. Our conclusion is that the tendons injected with the cytokine preparation represent a model of mild, seemingly reversible tendon injury. The cytokine preparation produces no matrix damage or evidence of collagen degradation and is species specific.

The effects of a popliteus muscle load on in situ forces in the posterior cruciate ligament and on knee kinematics. A human cadaveric study.

To investigate the effect of simulated contraction of the popliteus muscle on the in situ forces in the posterior cruciate ligament and on changes in knee kinematics, we studied 10 human cadaveric knees (donor age, 58 to 89 years) using a robotic manipulator/universal force moment sensor system. Under a 110-N posterior tibial load (simulated posterior drawer test), the kinematics of the intact knee and the in situ forces in the ligament were determined. The test was repeated with the addition of a 44-N load to the popliteus muscle. The posterior cruciate ligament was then sectioned and the knee was subjected to the same tests. The additional popliteus muscle load significantly reduced the in situ forces in the ligament by 9% to 36% at 90 degrees and 30 degrees of flexion, respectively. No significant effects on posterior tibial translation of the intact knee were found. However, in the ligament-deficient knee, posterior tibial translation was reduced by up to 36% of the translation caused by ligament transection. A coupled internal tibial rotation of 2 degrees to 4 degrees at 60 degrees to 90 degrees of knee flexion was observed in both the intact and ligament-deficient knees when the popliteus muscle load was added. Our results indicate that the popliteus muscle shares the function of the posterior cruciate ligament in resisting posterior tibial loads and can contribute to knee stability when the ligament is absent.

Determination of the in situ forces in the human posterior cruciate ligament using robotic technology. A cadaveric study.

We examined the in situ forces in the posterior cruciate ligament as well as the force distribution between its anterolateral and posteromedial bundles. Using a robotic manipulator in conjunction with a universal force-moment sensor system, we applied posterior tibial loads from 22 to 110 N to the joint at 0 degrees to 90 degrees of knee flexion. The magnitude of the in situ force in the posterior cruciate ligament and its bundles was significantly affected by knee flexion angle and posterior tibial loading. In situ forces in the posterior cruciate ligament ranged from 6.1 +/- 6.0 N under a 22-N posterior tibial load at 0 degree of knee flexion to 112.3 +/- 28.5 N under a 110-N load at 90 degrees. The force in the posteromedial bundle reached a maximum of 67.9 +/- 31.5 N at 90 degrees of knee flexion, and the force in the anterolateral bundle reached a maximum of 47.8 +/- 23.0 N at 60 degrees of knee flexion under a 110-N load. No significant differences existed between the in situ forces in the two bundles at any knee flexion angle. This study provides insight into the knee flexion angle at which each bundle of the posterior cruciate ligament experiences the highest in situ forces under posterior tibial loading. This information can help guide us in more accurate graft placement, fixation, and tensioning, and serve as an assessment of graft performance.

A functional comparison of animal anterior cruciate ligament models to the human anterior cruciate ligament.

Many investigators have used animal models to clarify the role of the human anterior cruciate ligament (ACL). Because none of these models are anatomically and biomechanically identical to the human ACL, there exists a need for an objective comparison of these models. To do this, we used a universal force-moment sensor to measure and compare the in situ forces, including magnitude and direction, of the ACL and the anteromedial (AM) and posterolateral (PL) bundles of human, pig, goat, and sheep knees. An Instron was used to apply 50 and 100 N anterior tibial loads at 90 degrees of knee flexion, while a universal force-moment sensor was used to measure the forces applied by the ACL to the tibia, the in situ force of the ACL. We found significant differences between the magnitude of force experienced by the goat and sheep ACL and AM and PL bundles when compared with the human ACL and AM and PL bundles. Also, the direction of the in situ force in the ACL and AM bundles of the goat and sheep were different from the human. The pig knee differed from the human only in the magnitude and direction of the in situ force in the PL bundle in response under anterior tibial loading. A tally of the significant differences between the animal models and the human knees indicates that goat and sheep knees may have limitations in modeling the human ACL, while the pig knee may be the preferred model for experimental studies.

Quantitative anthropometry of the subatlantal cervical longitudinal ligaments.

STUDY DESIGN: The quantitative anthropometry of the cervical longitudinal ligaments was determined in 20 human cadaveric subatlantal cervical spines at the limits of flexion and extension. OBJECTIVES: To provide measurements of cervical anterior and posterior longitudinal ligament lengths, widths, and cross-sectional areas at segmental levels. SUMMARY OF BACKGROUND DATA: Although mathematical models of the cervical spine require specific data to predict kinematics, the anthropometry of the cervical spine has not been examined in detail. The dimensional changes of ligaments in physiologic motion are not well characterized. METHODS: Segmental lengths and widths of the cervical longitudinal ligaments were measured in sagittal plane flexion and extension, using a three-dimensional electromagnetic digitizer. The cross-sectional areas of the ligaments at resting length were measured with a laser micrometer system. Comparisons between anterior and posterior location and among segmental levels were made. Several ligaments were examined histologically to determine the insertion sites and, thus, to define the segmental length. RESULTS: The anterior longitudinal ligaments were shorter in flexion than in extension. In extension, they were longer than the posterior longitudinal ligaments in flexion. The resting isolated ligaments were longer than the longest in situ lengths at several vertebral levels. The anterior longitudinal ligaments were wider at the disc than at the body. The cross-sectional area at C2-C3 was smaller than at subaxial levels. The longitudinal ligaments were observed to insert along the entire underlying vertebral body. CONCLUSIONS: The quantitative anthropometry of the cervical longitudinal ligaments is important in the development of accurate mathematical models of the cervical spine. The in situ ligaments may not be under tension in the physiologic range of motion.

Methotrexate and sulfasalazine promote adenosine release by a mechanism that requires ecto-5'-nucleotidase-mediated conversion of adenine nucleotides.

We and others have shown that an increased extracellular concentration of adenosine mediates the antiinflammatory effects of methotrexate and sulfasalazine both in vitro and in vivo, but the mechanism by which these drugs increase extracellular adenosine remains unclear. The results of the experiments reported here provide three distinct lines of evidence that adenosine results from the ecto-5'-nucleotidase- mediated conversion of adenine nucleotides to adenosine. First, pretreatment of a human microvascular endothelial cell line (HMEC-1) with methotrexate increases extracellular adenosine after exposure of the pretreated cells to activated neutrophils; the ecto-5'-nucleotidase inhibitor alpha, beta-methylene adenosine-5'-diphosphate (APCP) abrogates completely the increase in extracellular adenosine. Second, there is no methotrexate-mediated increase in extracellular adenosine concentration in the supernate of cells deficient in ecto-5'-nucleotidase, but there is a marked increase in extracellular adenosine concentration in the supernates of these cells after transfection and surface expression of the enzyme. Finally, as we have shown previously, adenosine mediates the antiinflammatory effects of methotrexate and sulfasalazine in the murine air pouch model of inflammation, and injection of APCP, the ecto-5'-nucleotidase inhibitor, abrogates completely the increase in adenosine and the decrement in inflammation in this in vivo model. These results not only show that ecto-5'-nucleotidase activity is a critical mediator of methotrexate- and sulfasalazine-induced antiinflammatory activity in vitro and in vivo but also indicate that adenine nucleotides, released from cells, are the source of extracellular adenosine.

In situ forces in the posterolateral structures of the knee under posterior tibial loading in the intact and posterior cruciate ligament-deficient knee.

The posterolateral structures of the knee consist of a complex anatomical architecture that includes several components with both static and dynamic functions. Injuries of the posterolateral structures occur frequently in conjunction with ruptures of the posterior cruciate ligament. To investigate the role of the posterolateral structures in maintaining posterior knee stability, we measured the in situ forces in the posterolateral structures and the distribution of force within the structures' major components, i.e., the popliteus complex and the lateral collateral ligament, in response to a posterior tibial load. Eight cadaveric knees were tested. With use of a robotic/universal force-moment sensor testing system, a posterior tibial load of 110 N was applied to the knee, and the resulting five-degree-of-freedom kinematics were measured at flexion angles of 0, 30, 60, 75, and 90 degrees. The knees were tested first in the intact state and then after the posterior cruciate ligament had been resected. These tests were also performed with an additional load of 44 N applied at the aponeurosis to simulate contraction of the popliteus muscle. In the intact knee, the in situ forces in the posterolateral structures were found to decrease with increasing knee flexion. After the posterior cruciate ligament was sectioned, these forces increased significantly at all angles of flexion. With no load applied to the popliteus muscle, the in situ forces in the popliteus complex were similar to those in the lateral collateral ligament. However, with a load of 44 N applied to the popliteus muscle, in situ forces in the popliteus complex were three to five times higher than those in the lateral collateral ligament. These results reveal that in response to posterior tibial loads, the posterolateral structures play an important role at full extension in intact knees and at all angles of flexion in posterior cruciate ligament-deficient knees. The popliteus muscle appears to be a major stabilizer under this loading condition; thus, the inability to restore its function may be a cause of unsatisfactory results in reconstructive procedures of the posterolateral structures of the knee.

Quantitative analysis of collagen fibrils of human cruciate and meniscofemoral ligaments.

The ultrastructural anatomy of collagen fibril diameters in the cruciate and meniscofemoral ligaments, from four young human cadaver knees (mean age, 20 years, range, 17-22 years) was studied using transmission electron microscopy. Samples were harvested from the proximal, middle, and distal regions of the anterior and posterior cruciate ligaments, and the meniscofemoral ligament. Photomicrographs were taken and assessed quantitatively using image analysis software to determine the collagen fibril diameters and eccentricities, and the percentage of total cross sectional area occupied by collagen. The collagen fibril diameter for the anterior cruciate ligament was found to be largest in the distal region but it decreased as it moved proximally. The posterior cruciate ligament had an opposite trend because it decreased from the proximal to the distal region. For the meniscofemoral ligament, the fibrils of the middle region were larger than those of the proximal and distal regions. The percentage of total cross sectional area occupied by collagen, however, did not vary significantly between regions. Fibril eccentricity also varied little between ligament or location. The variability observed in fibril diameters may account for the different mechanical properties of the ligaments.

Human anterior and posterior cervical longitudinal ligaments possess similar tensile properties.

One hundred bone-ligament-bone complexes were isolated from the cervical spines of 20 fresh-frozen human cadavers and mechanically tested in uniaxial tension. Load-elongation curves representing the structural properties of the complex were obtained, and stiffness, ultimate load, energy at failure, and elongation were determined from the curves. Stress-strain curves representing the mechanical properties of the ligament substance were also obtained, and elastic modulus was determined from these curves. Comparisons were made between anterior and posterior longitudinal ligaments and among cervical levels. The cross-sectional area of the anterior ligament was smaller at the C2-C3 level than at subaxial levels. There was a trend toward less stiffness but more energy at failure and greater elastic modulus for the anterior compared with the posterior ligaments at subaxial levels, although the differences were not statistically significant (p > 0.05).

Assessment of posterior cruciate ligament graft performance using robotic technology.

We used the information on in situ forces provided by robotics to compare two methods of posterior cruciate ligament graft fixation. Twenty porcine knees were studied using robotic technology to determine and repeat intact, deficient, and reconstructed knee motion under 110 N of posterior tibial loading at 30 degrees, 60 degrees, and 90 degrees of knee flexion. Reconstruction was performed using a bone-patellar tendon-bone graft with the distal end of the graft placed in the posterolateral aspect of the posterior cruciate ligament tibial insertion. Specimens were separated into two groups based on the femoral fixation site: the proximal or anterior aspect of the femoral insertion. Repetition of knee motion allowed measurement of the force in the intact posterior cruciate ligament and graft using the principle of superposition. The forces in the graft and the intact ligament provided additional information to evaluate graft performance. Force in the intact posterior cruciate ligament was significantly greater at 90 degrees than at 30 degrees and 60 degrees of knee flexion. The forces in both graft types were significantly lower than those of the posterior cruciate ligament, but the force in the anteriorly placed graft was significantly greater at 90 degrees than at 30 degrees and 60 degrees of knee flexion, similar to the intact posterior cruciate ligament. Thus, the anteriorly placed graft had a more physiologic increase in tension with knee flexion, when the joint provided less restraint.

The anti-inflammatory mechanism of sulfasalazine is related to adenosine release at inflamed sites.

The anti-inflammatory mechanism of sulfasalazine is not well understood. It has recently been shown that sulfasalazine inhibits 5-aminoimidazole-4-carboxamidoribonucleotide (AICAR) transformylase, an enzyme involved in de novo purine biosynthesis. We recently demonstrated that methotrexate promotes intracellular AICAR accumulation, thereby increasing adenosine release and diminishing inflammation, so we tested the hypothesis that sulfasalazine similarly promotes intracellular AICAR accumulation. We studied adenosine release and the state of inflammation in in vitro and in vivo models of the inflammatory process. The adhesion of stimulated neutrophils (FMLP) to endothelial cells preincubated with sulfasalazine was inhibited in a dose-dependent manner. Elimination of extracellular adenosine by addition of adenosine deaminase or inhibition of adenosine by the adenosine A2 receptor antagonist 3,7-dimethyl-1-propargylxanthine (DMPX) completely reversed the anti-inflammatory effect of sulfasalazine (at concentrations <1 microM in this in vitro model. To determine whether this phenomenon was relevant to inhibition of inflammation in vivo, we studied the effect of sulfasalazine (100 mg/kg/day by gastric gavage for 3 days) on leukocyte accumulation in the murine air pouch model of inflammation. Treatment with sulfasalazine markedly decreased the number of leukocytes that accumulated in the inflamed (carrageenan, 2 mg/ml) air pouch. Injection of either adenosine deaminase or DMPX, but not the A1 receptor antagonist 8-cyclopentyl-dipropylxanthine, significantly reversed the anti-inflammatory effects of sulfasalazine treatment. Sulfasalazine increased the exudate adenosine concentration from 127 +/- 64 nM to 869 +/- 47 nM. Moreover, sulfasalazine treatment promoted a marked increase in splenocyte AICAR concentration from 35 +/- 6 to 96 +/- 3 pmols/10(6) splenocytes, which is consistent with the in vitro observation that sulfasalazine inhibits AICAR transformylase. These results indicate that sulfasalazine, like methotrexate, enhances adenosine release at an inflamed site and that adenosine diminishes inflammation via occupancy of A2 receptors on inflammatory cells. Our studies provide evidence that sulfasalazine and methotrexate may be described as a newly recognized family of anti-inflammatory agents that share the property of using adenosine as an antagonist of inflammation.

In-situ forces in the human posterior cruciate ligament in response to posterior tibial loading.

Although some investigators have referred to the human posterior cruciate ligament (PCL) as the center of the knee, it has received less attention than the more frequently injured anterior cruciate ligament (ACL) and medial collateral ligament (MCL). Therefore, our understanding of the function of the PCL is limited. Our laboratory has developed a method of measuring the in-situ forces in a ligament without contacting that ligament by using a universal force-moment sensor (UFS). In this study, we attached a UFS to the tibia and measured in-situ forces of the human PCL as a function of knee flexion in response to tibial loading. At a 50-N posterior tibial load, the force in the PCL increased from 25 +/- 11 N (mean +/- SD) at 30 degrees of knee flexion to 48 +/- 12 N at 90 degrees of knee flexion. At 100 N, the corresponding increases were to 50 +/- 17 N and 95 +/- 17 N, respectively. Of note, at 30 degrees knee flexion, approximately 45% of the resistance to posterior tibial loading was caused by contact between the tibia and the femoral condyles, whereas, at 90 degrees of knee flexion, no resistance was caused by such contact. For direction of the in-situ force, the elevation angle from the tibial plateau was greater at 30 degrees of knee flexion than at 90 degrees of knee flexion. The data gathered on the magnitude and direction of the in-situ force of the PCL should help in our understanding of the dependence of knee flexion angle of the forces within the PCL.

The human posterior cruciate ligament complex: an interdisciplinary study. Ligament morphology and biomechanical evaluation.

To study the structural and functional properties of the human posterior cruciate ligament complex, we measured the cross-sectional shape and area of the anterior cruciate, posterior cruciate, and meniscofemoral ligaments in eight cadaveric knees. The posterior cruciate ligament increased in cross-sectional area from tibia to femur, and the anterior cruciate ligament area decreased from tibia to femur. The meniscofemoral ligaments did not change shape in their course from the lateral meniscus to their femoral insertions. The posterior cruciate ligament cross-sectional area was approximately 50% and 20% greater than that of the anterior cruciate ligament at the femur and tibia, respectively. The meniscofemoral ligaments averaged approximately 22% of the entire cross-sectional area of the posterior cruciate ligament. The insertion sites of the anterior and posterior cruciate ligaments were evaluated. The insertion sites of the anterior and posterior cruciate ligaments were 300% to 500% larger than the cross-section of their respective midsubstances. We determined, through transmission electron microscopy, fibril size within the anterior and posterior cruciate ligament complex from the femur to the tibia. The posterior cruciate ligament becomes increasingly larger from the tibial to the femoral insertions, and the anterior cruciate ligament becomes smaller toward the femoral insertion. We evaluated the biomechanical properties of the femur-posterior cruciate ligament-tibia complex using 14 additional human cadaveric knees. The posterior cruciate ligament was divided into two functional components: the anterolateral, which is taut in knee flexion, and the posteromedial, which is taut in knee extension. The anterolateral component had a significantly greater linear stiffness and ultimate load than both the posteromedial component and meniscofemoral ligaments. The anterolateral component and the meniscofemoral ligaments displayed similar elastic moduli, which were both significantly greater than that of the posteromedial component.

Anatomical and biomechanical characteristics of human meniscofemoral ligaments.

The meniscofemoral ligaments (MFL) of 26 human cadaver knees were studied to determine their structural importance. The incidence of at least one MFL in each of the specimens studied was 100%, and 46% of the specimens had both MFL ligaments (Humphry and Wrisberg). Another 23% had a single Humphry ligament, and the remaining 31% had a single Wrisberg ligament. A laser micrometer system was used to measure cross-sectional shape and area. The average cross-sectional areas of the Humphry and Wrisberg ligaments were 7.8 +/- 4.7 mm2 and 6.7 +/- 4.1 mm2, respectively. In specimens with both a Humphry and Wrisberg ligament, the larger ligament area was on average 100% greater than the smaller ligament area. The average ratios of the cross sectional areas of Wrisberg and Humphry to that for the PCL within the same knee were 12.0% +/- 7.7% and 11.9% +/- 5.7%, respectively. The structural properties of the MFL bone-ligament-meniscus complex and the mechanical properties of the MFL midsubstance were determined by uniaxial tensile testing. The average stiffness, ultimate load, and energy absorbed at failure were, respectively, 49.0 +/- 18.4 N/mm, 297.4 +/- 141.4 N and 1125.4 +/- 735.8 N/mm. The tangent modulus between 4% and 7% strain was 355.1 +/- 234.0 MPa. Our findings suggest that the MFL is a significant biomechanical structure in the knee because of its size, stiffness, and strength.

Sulphasalazine inhibition of human granulocyte activation by inhibition of second messenger compounds.

The effects of sulphasalazine on the production of second messenger compounds in human granulocytes have been characterised by various stimuli. The increases in cytosolic calcium, inositol trisphosphate, diacylglycerol, and phosphatidic acid (all important mediators of intracellular signal transduction) triggered by stimulation were inhibited by sulphasalazine. The metabolites 5-amino-salicylic acid and sulphapyridine were less potent inhibitors than the mother compound. It is concluded that sulphasalazine inhibits the synthesis of phosphoinositide derived second messenger compounds at the level of phospholipase C or its regulatory guanosine 5'-triphosphate (GTP) binding protein. Inhibition of phosphatidic acid synthesis was either due to the same mechanism, or to interaction with a phospholipase D regulating GTP binding protein.