We assessed in a piglet model the relationship between fecal ciprofloxacin concentrations and ciprofloxacin-resistant Enterobacteriaceae counts. Twenty-nine piglets were orally treated with placebo or with 1.5 or 15 mg ciprofloxacin/kg of body weight/day from day 1 (D1) to D5. Areas under the curve (AUC) of concentrations increased sharply with dose and correlated positively with AUC of resistant bacteria log counts between D1 and D9. Removing residual colonic quinolones could help to control the emergence of resistance in fecal flora.
Patients with the most common and aggressive form of high-grade glioma, glioblastoma multiforme, have poor prognosis and few treatment options. In 2 immunocompetent mouse brain tumor models (CT26-BALB/c and Tu-2449-B6C3F1), we showed that a nonlytic retroviral replicating vector (Toca 511) stably delivers an optimized cytosine deaminase prodrug activating gene to the tumor lesion and leads to long-term survival after treatment with 5-fluorocytosine (5-FC). Survival benefit is dose dependent for both vector and 5-FC, and as few as 4 cycles of 5-FC dosing after Toca 511 therapy provides significant survival advantage. In the virally permissive CT26-BALB/c model, spread of Toca 511 to other tissues, particularly lymphoid tissues, is detectable by polymerase chain reaction (PCR) over a wide range of levels. In the Tu-2449-B6C3F1 model, Toca 511 PCR signal in nontumor tissues is much lower, spread is not always observed, and when observed, is mainly detected in lymphoid tissues at low levels. The difference in vector genome spread correlates with a more effective antiviral restriction element, APOBEC3, present in the B6C3F1 mice. Despite these differences, neither strain showed signs of treatment-related toxicity. These data support the concept that, in immunocompetent animals, a replicating retroviral vector carrying a prodrug activating gene (Toca 511) can spread through a tumor mass, leading to selective elimination of the tumor after prodrug administration, without local or systemic pathology. This concept is under investigation in an ongoing phase I/II clinical trial of Toca 511 in combination with 5-FC in patients with recurrent high-grade glioma (www.clinicaltrials.gov NCT01156584).
brain cancer; cytosine deaminase; 5-fluorocyosine; gene therapy; glioblastoma multiforme
The literature reports inconsistent findings regarding the association between low back pain (LBP) and trunk muscle function, in both adults and children. The strength of the relationship appears to be influenced by how LBP is qualified and the means by which muscle function is measured. The aim of this study was to examine the association between isoinertial trunk muscle performance and consequential (non-trivial) low back pain (LBP) in male adolescents. Healthy male adolescents underwent anthropometric measurements, clinical evaluation, and tests of trunk range of motion (ROM), maximum isometric strength (STRENGTH) and peak movement velocity (VEL), using an isoinertial device. They provided information about their regular sporting activities, history and family history of LBP. Predictors of “relevant/consequential LBP” were examined using multivariable logistic regression. LBP status was reassessed after 2 years and the change from baseline was categorised. At baseline, 33/95 (35%) subjects reported having experienced consequential LBP. BMI, a family history of LBP, and regularly playing sport were each significantly associated with a history of consequential LBP (p < 0.05). 85/95 (89%) boys participated in the follow-up: 51 (60%) reported no LBP at either baseline or follow-up (never LBP); 5 (6%) no LBP at baseline, but LBP at follow-up (new LBP); 19 (22%) LBP at baseline, but none at follow-up; and 10 (12%) LBP at both time-points (recurrent/persistent LBP). The only distinguishing features of group membership in these small groups were: fewer sport-active in the “never LBP” group); worse trunk mobility, in the “persistent LBP” group, lower baseline sagittal ROM in the “never LBP” and “new LBP” (p < 0.05). Regular involvement in sport was a consistent predictor of LBP. Isoinertial trunk performance was not associated with LBP in adolescents.
Muscle strength; Range of motion; Sports; Adolescents; Low back pain
TIAR and HuR are mRNA-binding proteins that play important roles in the regulation of translation. They both possess three RNA recognition motifs (RRMs) and bind to AU-rich elements (AREs), with seemingly overlapping specificity. Here we show using SPR that TIAR and HuR bind to both U-rich and AU-rich RNA in the nanomolar range, with higher overall affinity for U-rich RNA. However, the higher affinity for U–rich sequences is mainly due to faster association with U-rich RNA, which we propose is a reflection of the higher probability of association. Differences between TIAR and HuR are observed in their modes of binding to RNA. TIAR is able to bind deoxy-oligonucleotides with nanomolar affinity, whereas HuR affinity is reduced to a micromolar level. Studies with U-rich DNA reveal that TIAR binding depends less on the 2′-hydroxyl group of RNA than HuR binding. Finally we show that SAXS data, recorded for the first two domains of TIAR in complex with RNA, are more consistent with a flexible, elongated shape and not the compact shape that the first two domains of Hu proteins adopt upon binding to RNA. We thus propose that these triple-RRM proteins, which compete for the same binding sites in cells, interact with their targets in fundamentally different ways.
Interspinous devices have been introduced to provide a minimally invasive surgical alternative for patients with lumbar spinal stenosis or foraminal stenosis. Little is known however, of the effect of interspinous devices on intersegmental range of motion (ROM). The aim of this in vivo study was to investigate the effect of a novel minimally invasive interspinous implant, InSwing®, on sagittal plane ROM of the lumbar spine using an ovine model. Ten adolescent Merino lambs underwent a destabilization procedure at the L1–L2 level simulating a stenotic degenerative spondylolisthesis (as described in our earlier work; Spine 15:571–576, 1990). All animals were placed in a side-lying posture and lateral radiographs were taken in full flexion and extension of the trunk in a standardized manner. Radiographs were repeated following the insertion of an 8-mm InSwing® interspinous device at L1–L2, and again with the implant secured by means of a tension band tightened to 1 N/m around the L1 and L2 spinous processes. ROM was assessed in each of the three conditions and compared using Cobb’s method. A paired t-test compared ROM for each of the experimental conditions (P < 0.05). After instrumentation with the InSwing® interspinous implant, the mean total sagittal ROM (from full extension to full flexion) was reduced by 16% from 6.3° to 5.3 ± 2.7°. The addition of the tension band resulted in a 43% reduction in total sagittal ROM to 3.6 ± 1.9° which approached significance. When looking at flexion only, the addition of the interspinous implant without the tension band did not significantly reduce lumbar flexion, however, a statistically significant 15% reduction in lumbar flexion was observed with the addition of the tension band (P = 0.01). To our knowledge, this is the first in vivo study radiographically showing the advantage of using an interspinous device to stabilize the spine in flexion. These results are important findings particularly for patients with clinical symptoms related to instable degenerative spondylolisthesis.
Interspinous implant; Biomechanics; Spondylolisthesis; Lumbar spine; Kinematics
Spinal musculature plays a major role in spine stability, but its importance to spinal stiffness is poorly understood. We studied the effects of graded trunk muscle stimulation on the in vivo dynamic dorsoventral (DV) lumbar spine stiffness of 15 adolescent Merino sheep. Constant voltage supramaximal electrical stimulation was administered to the L3–L4 interspinous space of the multifidus muscles using four stimulation frequencies (2.5, 5, 10, and 20 Hz). Dynamic stiffness was quantified at rest and during muscle stimulation using a computer-controlled testing apparatus that applied variable frequency (0.46–19.7 Hz) oscillatory DV forces (13-N preload to 48-N peak) to the L3 spinous process of the prone-lying sheep. Five mechanical excitation trials were randomly performed, including four muscle stimulation trials and an unstimulated or resting trial. The secant stiffness (ky = DV force/L3 displacement, kN/m) and loss angle (phase angle, deg) were determined at 44 discrete mechanical excitation frequencies. Results indicated that the dynamic stiffness varied 3.7-fold over the range of mechanical excitation frequencies examined (minimum resting ky = 3.86 ± 0.38 N/mm at 4.0 Hz; maximum ky = 14.1 ± 9.95 N/mm at 19.7 Hz). Twenty hertz muscle stimulation resulted in a sustained supramaximal contraction that significantly (P < 0.05) increased ky up to twofold compared to rest (mechanical excitation at 3.6 Hz). Compared to rest, ky during the 20 Hz muscle stimulation was significantly increased for 34 of 44 mechanical excitation frequencies (mean increase = 55.1%, P < 0.05), but was most marked between 2.55 and 4.91 Hz (mean increase = 87.5%, P < 0.05). For lower frequency, sub-maximal muscle stimulation, there was a graded change in ky, which was significantly increased for 32/44 mechanical excitation frequencies (mean increase = 40.4%, 10 Hz stimulus), 23/44 mechanical excitation frequencies (mean increase = 10.5%, 5 Hz stimulus), and 11/44 mechanical excitation frequencies (mean increase = 4.16%, 2.5 Hz stimulus) when compared to rest. These results indicate that the dynamic mechanical behavior of the ovine spine is modulated by muscle stimulation, and suggests that muscle contraction plays an important role in stabilizing the lumbar spine.
Biomechanics; Electromyography; Lumbar spine; Dynamic stiffness; Muscle stimulation
Gem is a protein of the Ras superfamily that plays a role in regulating voltage-gated Ca2+ channels and cytoskeletal reorganization. We now report that GTP-bound Gem interacts with the membrane–cytoskeleton linker protein Ezrin in its active state, and that Gem binds to active Ezrin in cells. The coexpression of Gem and Ezrin induces cell elongation accompanied by the disappearance of actin stress fibers and collapse of most focal adhesions. The same morphological effect is elicited when cells expressing Gem alone are stimulated with serum and requires the expression of ERM proteins. We show that endogenous Gem down-regulates the level of active RhoA and actin stress fibers. The effects of Gem downstream of Rho, i.e., ERM phosphorylation as well as disappearance of actin stress fibers and most focal adhesions, require the Rho-GAP partner of Gem, Gmip, a protein that is enriched in membranes under conditions in which Gem induced cell elongation. Our results suggest that Gem binds active Ezrin at the plasma membrane–cytoskeleton interface and acts via the Rho-GAP protein Gmip to down-regulate the processes dependent on the Rho pathway.
Spinal manipulation has been found to create demonstrable segmental and intersegmental spinal motions thought to be biomechanically related to its mechanisms. In the case of impulsive-type instrument device comparisons, significant differences in the force-time characteristics and concomitant motion responses of spinal manipulative instruments have been reported, but studies investigating the response to multiple thrusts (multiple impulse trains) have not been conducted. The purpose of this study was to determine multi-axial segmental and intersegmental motion responses of ovine lumbar vertebrae to single impulse and multiple impulse spinal manipulative thrusts (SMTs).
Fifteen adolescent Merino sheep were examined. Tri-axial accelerometers were attached to intraosseous pins rigidly fixed to the L1 and L2 lumbar spinous processes under fluoroscopic guidance while the animals were anesthetized. A hand-held electromechanical chiropractic adjusting instrument (Impulse) was used to apply single and repeated force impulses (13 total over a 2.5 second time interval) at three different force settings (low, medium, and high) along the posteroanterior axis of the T12 spinous process. Axial (AX), posteroanterior (PA), and medial-lateral (ML) acceleration responses in adjacent segments (L1, L2) were recorded at a rate of 5000 samples per second. Peak-peak segmental accelerations (L1, L2) and intersegmental acceleration transfer (L1–L2) for each axis and each force setting were computed from the acceleration-time recordings. The initial acceleration response for a single thrust and the maximum acceleration response observed during the 12 multiple impulse trains were compared using a paired observations t-test (POTT, alpha = .05).
Segmental and intersegmental acceleration responses mirrored the peak force magnitude produced by the Impulse Adjusting Instrument. Accelerations were greatest for AX and PA measurement axes. Compared to the initial impulse acceleration response, subsequent multiple SMT impulses were found to produce significantly greater (3% to 25%, P < 0.005) AX, PA and ML segmental and intersegmental acceleration responses. Increases in segmental motion responses were greatest for the low force setting (18%–26%), followed by the medium (5%–26%) and high (3%–26%) settings. Adjacent segment (L1) motion responses were maximized following the application of several multiple SMT impulses.
Knowledge of the vertebral motion responses produced by impulse-type, instrument-based adjusting instruments provide biomechanical benchmarks that support the clinical rationale for patient treatment. Our results indicate that impulse-type adjusting instruments that deliver multiple impulse SMTs significantly increase multi-axial spinal motion.
The problems linked to blood loss and blood-sparing techniques in spine surgery have been less studied than in other fields of orthopedics, such as joint-replacement procedures. Decreasing bleeding is not only important for keeping the patient’s hemodynamic equilibrium but also for allowing a better view of the surgical field. In spine surgery the latter aspect is especially important because of the vicinity of major and highly fragile neurologic structures. The techniques and agents used for hemostasis and blood sparing in spinal procedures are mostly similar to those used elsewhere in surgery. Their use is modulated by the specific aspects of spinal approach and its relation to the contents of the spinal canal. Blood-sparing techniques can be divided into two categories based on their goals: either they are aimed at decreasing the bleeding itself, or they are aimed at decreasing the need for homologous transfusion. Various hemodynamic techniques, as well as systemic and local drugs and agents, can be used separately or in combination, and their use in the field of spine surgery is reported. The level of evidence for the efficacy of many of those methods in surgery as a whole is limited, and there is a lack of evidence for most of them in spine surgery. However, several blood-saving procedures and drugs, as well as promising new agents, appear to be efficient, although their efficacy has yet to be assessed by proper randomized controlled trials.
Hemostasis; Spine surgery; Bleeding; Blood sparing; Prevention
This study was designed to develop predictive models for surgical outcome based on information available prior to lumbar stenosis surgery. Forty patients underwent decompressive laminarthrectomy. Preop and 1-year postop evaluation included Waddell’s nonorganic signs, CT scan, Waddell disability index, Oswestry low back pain disability questionnaire, low back outcome score (LBOS), visual analog scale (VAS) for pain intensity, and trunk strength testing. Statistical comparisons of data used adjusted error rates within families of predictors. Mathematical models were developed to predict outcome success using stepwise logistic regression and decision-tree methodologies (chi-squared automatic interaction detection, or CHAID). Successful outcome was defined as improvement in at least three of four criteria: VAS, LBOS, and reductions in claudication and leg pain. Exact logistic regression analysis resulted in a three-predictor model. This model was more accurate in predicting unsuccessful outcome (negative predictive value 75.0%) than in successful outcome (positive predictive value 69.6%). A CHAID model correctly classified 90.1% of successful outcomes (positive predictive value 85.7%, negative predictive value 100%). The use of conservative surgical decompression for lumbar stenosis can be recommended, as it demonstrated a success rate similar to that of more invasive techniques. Given its physiologic and biomechanical advantages, it can be recommended as the surgical method of choice in this indication. Underlying subclinical vascular factors may be involved in the complaints of spinal stenosis patients. Those factors should be investigated more thoroughly, as they may account for some of the failures of surgical relief. The CHAID decision tree appears to be a novel and useful tool for predicting the results of spinal stenosis surgery
Biomechanics; Low back pain; Lumbar stenosis; Outcome scales; Predictive models