Management of recalcitrant osteoarthritis of the atlanto-axial joint.

The medical records and radiographs of 17 patients with recalcitrant pain secondary to C1-C2 osteoarthritis were independently reviewed. All patients were treated by the senior author (K.D.R.). Initial treatment included physical therapy, nonsteroidal anti-inflammatory drugs, and soft collar immobilization. If non-responsive, patients were referred to a single radiologist for injection of the involved C1-C2 joint(s). If symptoms persisted, then transarticular screw fixation and Magerl wire fixation with structural iliac crest bone graft was performed. Postoperatively, all patients were placed in a soft collar. Postoperative anteroposterior (AP), lateral, flexion/extension, and open-mouthed odontoid radiographs were taken at 6 weeks, 3 months, and 1 year. Three independent observers assessed all radiographs for fusion. Fusion was noted by radiographic evidence of bridging bony trabeculae across the C1-C2 joint and/or bridging bone from the posterior arch of C1 to the C2 spinous process. Patient satisfaction improved significantly whether they were treated by corticosteroid injection or surgically.

Comparison of manual and digital measurements in adolescent idiopathic scoliosis.

STUDY DESIGN: Comparison of manual and digital measurement of radiographic parameters in patients with adolescent idiopathic scoliosis (AIS). OBJECTIVE: To assess the reliability of digital measures as compared to manual measures in the evaluation of AIS. SUMMARY OF BACKGROUND DATA: Radiographic parameters are critical to the evaluation of patients with AIS, and are frequently used to monitor curve progression and guide treatment decisions. The reliability of many of the more common radiographic measures has only recently been elucidated for both manual and digital measures. However, a comparative analysis of manual versus digital measures has been performed only for coronal Cobb angles. The inter-technique reliability of these parameters will have increasing importance as digital radiographic viewing and analysis become commonplace. METHODS: There were 2 independent, blinded observers that measured 30 complete sets of preoperative (posterior-anterior, lateral, and both side-bending) and postoperative (posterior-anterior and lateral) radiographs on 4 different occasions. For the first 2 iterations, manual measurements were taken using the same pencil and protractor. For the last 2 iterations, measurements of digitized radiographs were taken on a software measurement program (PhDx, Albuquerque, NM). Coronal measures included the main thoracic and thoracolumbar/lumbar standing and side-bending Cobb angles, apical vertebral translation, coronal balance, T1 tilt angle, lowest instrumented vertebrae angle, angulation of the disc inferior to the lowest instrumented vertebrae, apical Nash-Moe vertebral rotation, and Risser grade. Sagittal parameters included T2-T5 and T5-T12 regional thoracic kyphosis, T2-T12 thoracic kyphosis, T10-L2 thoracolumbar junction sagittal curvature, T12-S1 lumbar lordosis, and global sagittal balance. The technique-dependent measurement variability and the inter-technique (manual vs. digital), intraobserver reliability were evaluated for each radiographic parameter (within 3 degrees ). RESULTS: Digital measurement showed decreased intraobserver variability for many (9 of 15) of the radiographic parameters assessed. Likewise, digital measures indicated good or excellent correlation with the absolute values obtained with manual measurement for many (10 of 15) parameters. All but 1 of those parameters having moderate-to-poor correlation had been previously shown to have poor reliability, regardless of measurement technique. Statistically significant differences between measurement variability were noted for 6 measures, including 2 favoring digital and 4, manual. Significant differences in the absolute values were noted for 5 measures, determined at a difference of 3 degrees . However, the differences in both parameter variability and absolute values tended to be small and of little clinical significance for manual versus digital measurement. CONCLUSIONS: Digital measurement showed improved measurement precision and good correlation with manual measurements for the majority of AIS parameters. Absolute differences between manual and digital measurements were generally small. Therefore, digital measures are acceptable as a valid technique for scoliosis evaluation. The importance of digital versus manual measurement reliability will increase as digital radiographic viewing becomes more prevalent.

Use of fluoroscopy to evaluate iliac screw position.

Iliac screw fixation is often used for long fusions to the sacropelvis. Maximum iliac screw purchase is obtained both by placing the screws within 1.5 cm of the greater sciatic notch and by extending them anterior to the axis of rotation in flexion-extension. Screw insertion is "blinded" or dependent on tactile feedback, and hence extreme care is necessary to avoid incorrect placement and damage to vital neurovascular structures in the pelvis and sciatic notch. Long screws may violate the hip joint while medial placement may injure the lumbosacral plexus and the nearby vessels. To explore the best intraoperative fluoroscopic method of determining optimal iliac screw placement, we used a synthetic pelvis model to investigate screw placement conditions: (1) optimal anatomic placement, (2) violation of the sciatic notch, (3) hip joint violation, (4) medial wall violation, and (5) lateral wall violation. Each condition was examined utilizing fluoroscopy with posteroanterior, inlet, outlet, lateral, iliac oblique, and obturator oblique Judet views to simulate operative conditions. These views were obtained to evaluate critical malposition of iliac screws. We found that, for a sciatic notch violation, the obturator oblique view best demonstrated the cortical breech, while for a hip joint violation, the inlet and outlet views were best. For a medial wall violation, the iliac oblique view best showed the violation. For a lateral wall violation, we were unable to demonstrate the cortical breech using these fluoroscopic views. Fluoroscopy is an effective method to determine sciatic notch, hip joint, and medial wall violations after iliac screw placement; however, it is not effective in identifying a lateral wall violation.

Reliability of end, neutral, and stable vertebrae identification in adolescent idiopathic scoliosis.

STUDY DESIGN: Analysis of radiographic interpretation and vertebral level identification. OBJECTIVES: To assess the intra- and interobserver reliability by observer training level used for selecting the end vertebra (EV), neutral vertebra (NV), and stable vertebra (SV) in adolescent idiopathic scoliosis patients. SUMMARY OF BACKGROUND DATA: Various radiographic and clinical factors are important in surgical planning. For adolescent idiopathic scoliosis, an analysis of the end, neutral, and stable vertebrae are of paramount importance for understanding spinal deformity management and determining the distal fusion level. Additionally, the development and comparison of optimal surgical techniques requires reliable, reproducible radiographic parameters. METHODS: One hundred consecutive radiographs of operative cases of adolescent idiopathic scoliosis were evaluated on three separate occasions by three surgeons (2700 data points) at various levels of training (fellowship-trained spine surgeon, fellow in-training, orthopedic surgery resident). For each iteration, the observers attempted to identify the distal structural Cobb curve EV, NV, and SV. The radiographs included preselected Lenke type 1, 3, and 5 curves in random order. The average main thoracic curve was 53 degrees (range, 30-82 degrees) with a T8-T9 average apex, whereas the average thoracolumbar curve was 33 degrees (range, 18-65 degrees). Intra- and interobserver reliability was assessed by means of Cohen's Kappa correlation coefficient, and raw percentages of agreement were recorded. RESULTS: Intraobserver reliability was good to excellent for determining the EV (kappaa = 0.69-0.88), good for determining the NV (kappaa = 0.65-0.73), and good to excellent for determining the SV (kappaa = 0.74-0.91) with 83.5, 72.2, and 85.6% intraobserver agreement, respectively. A trend was noted towards greater intraobserver reliability with increasing levels of observer experience. Interobserver reliability was poor (kappaa = 0.26-0.39) for each vertebral level, with interobserver agreement for only 48.7% of EV, 41.7% of NV, and 51.0% of SV. However, interobserver agreement increased significantly when concurrence within one vertebral level was assessed, with 91, 73, and 76% agreement for identifying the EV, NV, and SV, respectively. CONCLUSIONS: Radiographic determination of the EV, NV, and SV demonstrated good to excellent intraobserver, but poor interobserver, reliability. Interobserver agreement was fair to good when concurrence within one adjacent level was assessed. Observer experience level may be a factor. The difficulties in identifying these vertebral levels represent a potential obstacle to reproducible patient-specific fusion level determination and to the optimization and uniformity of patient care.

Comparison of sagittal contour and posterior disc height following interbody fusion: threaded cylindrical cages versus structural allograft versus vertical cages.

OBJECTIVE: Segmental restoration of sagittal contour is recognized as critical for improved long-term success following instrumented lumbar fusions. As such, the use of wedged implants has become more popular. Few studies exist to assess the postoperative lordotic and disc height changes following these varied techniques in spinal fusion. An observational radiographic study examining lumbar sagittal contour and posterior intervertebral disc space height following posterior lumbar interbody fusion (PLIF) or transforaminal lumbar interbody fusion (TLIF) was conducted using vertical cages (VCs), wedged structural allograft (WSA), and threaded cylindrical cages (TCCs). METHODS: Forty-nine consecutive patients (59 spinal segments) were evaluated following single- or two-level interbody fusion with either stand-alone TCCs (n = 18 levels), WSA with posterior transpedicular compression instrumentation (n = 25 levels), or VCs with posterior transpedicular compression instrumentation (n = 16 levels). Standing lumbar radiographs were measured by two independent observers preoperatively, immediately postoperatively (within 1 week), at 6-week follow-up (range 4-8 weeks), and postoperatively (at 1-year follow-up) for segmental lordosis at each level undergoing posterior interbody arthrodesis and posterior intervertebral disc space height to assess indirect nerve root decompression. RESULTS: At the 1-year follow-up, postoperative lordosis was improved in the VC group (+5.3 degrees ; P < 0.005), whereas it decreased in the WSA group (-0.9 degrees ; P = 0.407) and TCC group (-3.5 degrees ; P < 0.005). The posterior disc space height decreased in the VC group (-0.5 mm; P = 0.109), whereas it increased for both the WSA group (+1.2 mm; P = 0.05) and the TCC group (+0.8 mm; P = 0.219). CONCLUSIONS: PLIF with stand-alone TCC and PLIF (or TLIF) with WSA and posterior transpedicular instrumentation results in an increased posterior disc height and thus improved indirect nerve root decompression. PLIF (or TLIF) with VC and posterior transpedicular instrumentation results in an overall decrease in posterior disc height. However, TCC and WSA resulted in a loss of lumbar lordosis, whereas VC resulted in an increase in lumbar lordosis.

Reliability analysis for digital adolescent idiopathic scoliosis measurements.

OBJECTIVE: Analysis of adolescent idiopathic scoliosis (AIS) requires a thorough clinical and radiographic evaluation to completely assess the three-dimensional deformity. Recently, these radiographic parameters have been analyzed for reliability and reproducibility following manual measurements; however, most of these parameters have not been analyzed with regard to digital measurements. The purpose of this study is to determine the intra- and interobserver reliability of common scoliosis radiographic parameters using a digital software measurement program. METHODS: Thirty sets of preoperative (posteroanterior [PA], lateral, and side-bending [SB]) and postoperative (PA and lateral) radiographs were analyzed by three independent observers on two separate occasions using a software measurement program (PhDx, Albuquerque, NM). Coronal measures included main thoracic (MT) and thoracolumbar-lumbar (TL/L) Cobb, SB MT Cobb, MT and TL/L apical vertical translation (AVT), C7 to center sacral vertical line (CSVL), T1 tilt, LIV tilt, disk below lowest instrumented vertebra (LIV), coronal balance, and Risser, whereas sagittal measures included T2-T5, T5-T12, T2-T12, T10-L2, T12-S1, and sagittal balance. Analysis of variance for repeated measures or Cohen three-way kappa correlation coefficient analysis was performed as appropriate to calculate the intra- and interobserver reliability for each parameter. RESULTS: The majority of the radiographic parameters assessed demonstrated good or excellent intra- and interobserver reliability. The relationship of the LIV to the CSVL (intraobserver kappaa = 0.48-0.78, fair to excellent; interobserver kappaa = 0.34-0.41, fair to poor), interobserver measurement of AVT (rho = 0.49-0.73, low to good), Risser grade (intraobserver rho = 0.41-0.97, low to excellent; interobserver rho = 0.60-0.70, fair to good), intraobserver measurement of the angulation of the disk inferior to the LIV (rho = 0.53-0.88, fair to good), apical Nash-Moe vertebral rotation (intraobserver rho = 0.50-0.85, fair to good; interobserver rho = 0.53-0.59, fair), and especially regional thoracic kyphosis from T2 to T5 (intraobserver rho = 0.22-0.65, poor to fair; interobserver rho = 0.33-0.47, low) demonstrated lesser reliability. In general, preoperative measures demonstrated greater reliability than postoperative measures, and coronal angular measures were more reliable than sagittal measures. CONCLUSIONS: Most common radiographic parameters for AIS assessment demonstrated good or excellent reliability for digital measurement and can be recommended for routine clinical and academic use. Preoperative assessments and coronal measures may be more reliable than postoperative and sagittal measurements. The reliability of digital measurements will be increasingly important as digital radiographic viewing becomes commonplace.

Un nuevo aporte a la toxicología de colorantes alimentarios: conjugación hepática de la indigotina con fosfolípidos / A new approach to the toxicology of food colorants: hepatic conjugation of indigotin with phospholipids

Demostramos anteriormente que la solubilidad del colorante alimentario indigotina se modifica en el hígado de mamíferos, volviéndose totalmente insoluble en solventes acuosos y altamente soluble en solventes orgánicos. En el presente trabajo se estudió la movilidad cromatográfica de la indigotina presente en un extracto lipídico de hígado de rata, 20 minutos después de su administración intravenosa. Se estudió además la presencia de grupos funcionales correspondientes a su probable conjugación con fosfolípidos, en el extracto preparado al raspar la banda del colorante separada mediante cromatografía de fosfolípidos en capa delgada. Se detectaron 0,144 +/- 0,087 µg de fósforo fosfolipídico y se demostró la presencia de ácidos grasos en dicha fracción, proveniente de un extracto preparado a partir de 2 gramos de hígado. Estos resultados confirman el aumento en la solubilidad del colorante en solventes orgánicos una vez que ha ingresado al hígado y sugieren que el mismo sufre una probable conjugación con fosfolípidos en dicho órgano.

We have demonstrated that the solubility of the food colorant indigotin was modified in mammalian liver, where it becomes totally insoluble in aqueous solvents and highly soluble in organic ones. This work presents the results from the studies on the chromatographic mobility of indigotin present in the lipidic extract obtained from rat liver 20 minutes after its intravenously administration. The presence of functional groups corresponding to its probable conjugation with phospholipids was also studied in the extract prepared from the colorant spot separated by thin layer chromatography of phospholipids. 0,144 +/- 0,087 _g of phospholipidic phosphorus were detected and fatty acid presence was demosntrated in this fraction, obtained from liver extract prepared with 2 grams of tissue. These results confirm the increment of the colorant solubility in organic solvents once it is incorporated in the liver and they suggest that it suffers a probably conjugation with phospholipids in this organ.

Reliability analysis for manual adolescent idiopathic scoliosis measurements.

STUDY DESIGN: Manual radiographic measurement analysis. OBJECTIVES: To determine the intraobserver and interobserver reliability of numerous radiographic process measures used in the assessment of adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Analysis of scoliosis requires a thorough radiographic evaluation to completely assess the deformity. Numerous radiographic process measures have been studied extensively and used for outcomes assessment and thus become the de facto standard of care. However, many of these measures have not been evaluated to determine the reliability and reproducibility. Validation of radiographic process measures is necessary to compare these measures with patient-focused outcome measures, as well as to permit valid comparison of different surgical techniques. METHODS: Thirty complete sets of long-cassette scoliosis radiographs (anteroposterior [AP], lateral and side-bending preoperative and AP, and lateral postoperative) were analyzed by three independent experienced observers on two separate occasions. Coronal image measures included the coronal Cobb angles, side-bending Cobb, apical vertebral translation, coronal balance, T1 tilt, lowest instrumented vertebrae (LIV) tilt, angulation of the disc below the LIV, apical vertebral rotation (Nash-Moe),and Risser sign; sagittal measures included T2-T5, T5-T12, T2-T12, T10-L2, T12-S1, and sagittal balance. Intraobserver and interobserver reliability for each measure was then assessed. RESULTS: The vast majority of the radiographic process measures assessed demonstrated good to excellent or excellent intraobserver and interobserver reliability. However, the angulation of the disc below the LIV demonstrated only fair interobserver reliability for postoperative measurements (rho = 0.59). Likewise, Risser grade measurements reflected good intraobserver (0.81-0.99) but only fair interobserver reliability (0.60-0.70). Apical vertebral rotation assessed by the technique of Nash and Moe produced good intraobserver reliability before surgery (0.74-0.85) but only fair reliability after surgery (0.50-0.85). The interobserver reliability for apical Nash-Moe rotation was fair to poor (0.53-0.59). For T2-T5 regional kyphosis, intraobserver (0.22-0.83) and interobserver (0.33-0.47) reliability was generally poor. Overall, the reliability of postoperative measurements tended to be decreased relative to preoperative values, likely due to instrumentation overlying radiographic landmarks. CONCLUSIONS: Most of the radiographic process measures evaluated in this study demonstrated good or excellent reliability. The reliability of measuring the angulation of the disc below the LIV, the apical Nash-Moe rotation, and Risser grading was decreased relative to other measures. The reliability of measuring T2-T5 regional kyphosis was disappointing and poor. With regards to the other 13 measures assessed, our findings support the use of these process measures obtained by experienced deformity surgeons via manual measurement for routine clinical and academic purposes.

Accuracy and efficacy of thoracic pedicle screws in curves more than 90 degrees.

STUDY DESIGN: Retrospective study of large-magnitude thoracic curves (> or =90 degrees ) treated with pedicle screw constructs. OBJECTIVE: To evaluate the results of pedicle screw constructs for thoracic curves > or = 90 degrees in terms of sagittal and coronal correction/efficacy, as well as accuracy and safety of thoracic pedicle screw placement. SUMMARY OF BACKGROUND DATA: Thoracic pedicle screw constructs continue to become increasingly more common; however, the debate continues about the safety and efficacy of these constructs because of their perceived increased risk of neurologic injury and the increased cost of spinal instrumentation. METHODS: Since 1998, all patients with adolescent idiopathic scoliosis, or adult progression of adolescent idiopathic scoliosis, a thoracic curve > or = 90 degrees and a minimum 2-year follow-up who were treated with pedicle screw constructs were included in this study. Standing anteroposterior (or posteroanterior), lateral and bending preoperative radiographs, and anteroposterior (or posteroanterior) and lateral postoperative radiographs were evaluated for curve magnitude, flexibility, and postoperative correction to assess the efficacy of these constructs in the immediate postoperative period and at latest follow-up. Postoperative CT scans were evaluated for screw accuracy using established 2-mm increments (intrapedicular, 0-2 mm breach, 2-4 mm breach, > 4 mm breach). Preoperative plans were also reviewed to evaluate the ability to place a pedicle screw at each planned level in these large-magnitude curves. RESULTS: Twenty patients with thoracic curves > or = 90 degrees and an average follow-up of 3.3 years (range, 2.0-5.2 years) were included in the study. All patients underwent a posterior spinal fusion with a pedicle screw only construct. The average preoperative main thoracic curve measured 100.2 degrees (range, 90 degrees -133 degrees ), with an average side-bender of 71.6 degrees (29% flexibility). The average postoperative main thoracic curve was 32.3 degrees (68% correction). A total of 352 thoracic screws were placed in the 20 cases (17.6 screws/case). Screw accuracy (either intrapedicular or <2 mm breach) was 96.3% (339 of 352 screws) by postoperative CT scanning. Ten screws were considered to have a breach between 2 and 4 mm (3 medial, 7 lateral), while three screws were > 4 mm (2 medial, 1 lateral). The two medial screws were the only placed screws that were removed (0.57%). Overall, 94% of planned screws (352 of 374 screws) were placed according to the preoperative plan. There were no incidences of screw or instrumentation failure. Of note, there was a temporary decrease in motor-evoked potentials during curve correction in 2 cases; however, there were no identifiable neurologic complications. CONCLUSIONS: Thoracic pedicle screw constructs can be safely used for large-magnitude curves. Curve correction (68%) is powerful for these curves, which are stiff and difficult to manage. Correction should be performed carefully with consideration given to convex compression for cases with concomitant hyperkyphosis for these "at risk" spinal cords. Screw accuracy (96.3%) was excellent in this review. The authors found that screws can consistently be placed according to the preoperative plan even in these large-magnitude curves.

The effect of Zn2+ on glucose 6-phosphate dehydrogenase activity from Bufo arenarum toad ovary and alfalfa plants.

The effect of Zn2+ on glucose 6-phosphate dehydrogenase (G6PD) activity was monitored in samples from Bufo arenarum toad ovary and alfalfa plants, in the search for a possible new bioindicator able to detect levels of exposure through contaminated soils, and also to elucidate possible similarities between the enzyme from animal and plant tissues. The in vivo effect was evaluated after exposure of the toads to the metal in Ringer solution during 30 days and after 10 days of treatment in 6 weeks old plants, cultured under laboratory conditions. In vitro effects were measured in different extracts from control samples and partially purified enzyme from ovarian tissue as well as in different extracts from control alfalfa plants, by addition of the metal to the reaction mixture containing the enzyme. G6PD from toad ovary was noncompetitively inhibited by zinc both in vivo and in vitro, under all the experimental conditions studied. A kinetic analysis of the enzyme activity showed that the Michaelis-Menten constant (Km) was not modified, while maximal velocity (Vmax) decreased as the consequence of treatment. It was not possible to obtain a dose-response curve for the effects of Zn2+ on G6PD from alfalfa whole plants, measured in vivo or in vitro. Only leaf extracts evidenced a possible relationship between treatment with the metal and G6PD activity alteration. The results agree with a possible role for G6PD as a biomarker of effect and exposure to Zn2+ in B. arenarum ovarian tissue but not in alfalfa plants.

Effects of long-term exposure to Cu2+ and Cd2+ on the pentose phosphate pathway dehydrogenase activities in the ovary of adult Bufo arenarum: possible role as biomarker for Cu2+ toxicity.

The effects of copper and cadmium on metabolism through the pentose phosphate pathway were evaluated in Bufo arenarum toad ovary. The effects of the two metals on dehydrogenases from this pathway were evaluated by three experiments: (1) in samples obtained from control females with addition of the metals to the reaction mixture (in vitro), (2) in samples obtained from control females and after long-term exposure of females to 4 and 100 microg/L of Cu or Cd in the incubation media (in vitro after exposure to the metals in vivo), and (3) 14CO2 production through the pentose phosphate pathway was evaluated after [U-14C]glucose microinjection on ovulated oocytes (in vivo after microinjection of the metals). Results from (1) evidenced inhibition of both enzyme activities but only above 1.5 mM Cu and Cd added to the reaction mixture. In (2) both glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities decreased in samples from the ovaries of females exposed in vivo to Cu, in a concentration-dependent manner (up to 90% in females exposed to 100 microg/L Cu: 2.12 +/- 1.57 NADPH micromol/min microg protein x 10(-5) vs 19.97 +/- 8.54 in control females). Cd treatment of the toads only rendered an inhibitory effect on 6-phosphogluconate dehydrogenase activity after exposure to 4 microg/L of the bivalent cation. (3) In vivo 14CO2 evolution significantly decreased in oocytes coinjected with 6.3 x 10(-3) mM Cu (calculated intracellular final concentration of the metal injected) and radioactive glucose. Cu and Cd concentration in samples from exposed females were always under detection limit by particle-induced X-ray emission. The results presented here are in agreement with a role for both glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities determination as biomarkers of effect and exposure for Cu but not for Cd toxicity.

Anterior thoracic scoliosis constructs: effect of rod diameter and intervertebral cages on multi-segmental construct stability.

BACKGROUND CONTEXT: Many studies have reported on the use of anterior instrumentation for thoracolumbar scoliosis and more recently thoracic scoliosis. However, the optimal construct design remains an issue of debate. PURPOSE: To optimize construct design and enhance implant survival until a successful spinal arthrodesis is achieved. STUDY DESIGN: This study evaluated the effect of rod diameter and intervertebral cages on construct stiffness and rod strain using a long-segment, anterior thoracic scoliosis model with varying levels of intervertebral reconstruction. METHODS: Sixteen fresh-frozen calf spine specimens (T1 to L1) were divided into two groups based on rod diameter reconstruction (4 mm and 5 mm). Testing included axial compression, anterior flexion, extension and lateral bending with variations in the number and level of intervertebral cage reconstructions: apical disc (one), end discs (two), apical and end discs (three), all seven levels (seven). Multisegmental construct stiffness and rod strain were determined and normalized to the intact specimen for analysis. RESULTS: The seven-level intervertebral cage construct showed significantly greater stiffness in axial compression for both the 4-mm (366% increased stiffness) and 5-mm (607% increased stiffness) rod groups (p<.001). The remaining constructs were not significantly different from each other (p>.05). In flexion, similar results were obtained for the 4-mm construct (p<.001) but not the 5-mm construct, because the reconstruction-alone, one-, two- and three-cage constructs were all significantly stiffer than the intact specimen (p<.05). Multisegmental construct stiffness under extension loading, as well as right and left lateral bending, also exhibited significant differences between the seven-level interbody cage reconstructions and the remaining constructs. Apical rod strain for both the 4-mm-rod and 5-mm-rod groups were significantly higher for the two cage constructs (a cage at either end but not the apex where the strain gauges were located) as compared with the other constructs (p<.05). These differences were more pronounced in the 4-mm-rod group. Similar results were obtained in anterior flexion, extension and lateral bending. CONCLUSIONS: Intervertebral cages at every level significantly improved construct stiffness compared with increasing rod diameter alone. Moreover, cages markedly decreased rod strain, and when structural interbody supports were not used, axial compression created the greatest rod strain.

Mecanismos de acción de xenobióticos en ovocitos de anfibios / Xenobiotic mechanisms of action in amphibian oocytes

Comprender los procesos que determinan la toxicidad de diversas sustancias a nivel celular constituye un desafío para el desarrollo tecnológico actual. Los ovocitos de diversas especies son utilizados con distintos propósitos en investigación. El tamaño de los ovocitos de anfibios y la facilidad con que puede obtenerse un gran número de ellos como células aisladas, hacen de los mismos el sistema de elección para múltiples propósitos, tales como: estudios de las etapas tempranas de la reproducción, técnicas de fertilización in vitro, expresión y caracterización de receptores de membranas plasmática, mecanismos de acción de agonistas, vías metabólicas, etc. El objetivo del presente trabajo con ovocitos de sapo Bufo arenarum es estudiar los mecanismos de acción de sustancias xenobióticas a nivel bioquímico y molecular, en relación con la capacidad de éstas gametas de ser fertilizadas y avanzar a través de los estadios tempranos del desarrollo embrionario. Se ha trabajado con diversos biomarcadores que permitieron reconocer los mecanismos de toxicidad de compuestos organoclorados y más recientemente de algunos metales pesados. Se demostró que la activación de una fosfolipasa C específica por el Dieldrin provoca depleción de mediadores lipídicos y como consecuencia inhibición de la fertilización (desarrollo hasta 4-8 células). Actualmente se estudian los efectos del Zn sobre la fertilización y su probable relación con alteraciones en la actividad de enzimas de la vía de las pentosas, directamente ligada a los mecanismos de detoxificación en presencia de stress oxidativo. El estudio de los mecanismos de citotoxicidad de diversas sustancias resulta de gran utilidad cuando se lleva a cabo en gametas, porque permite también evaluar los efectos de dichas sustancias sobre la reproducción de la especie. Los ovocitos de anfibios son además células de gran interés, por su utilidad en el estudio de la citotoxicidad de una amplia variedad de contaminantes hídricos

Mecanismos de acción de xenobióticos en ovocitos de anfibios / Xenobiotic mechanisms of action in amphibian oocytes

Comprender los procesos que determinan la toxicidad de diversas sustancias a nivel celular constituye un desafío para el desarrollo tecnológico actual. Los ovocitos de diversas especies son utilizados con distintos propósitos en investigación. El tamaño de los ovocitos de anfibios y la facilidad con que puede obtenerse un gran número de ellos como células aisladas, hacen de los mismos el sistema de elección para múltiples propósitos, tales como: estudios de las etapas tempranas de la reproducción, técnicas de fertilización in vitro, expresión y caracterización de receptores de membranas plasmática, mecanismos de acción de agonistas, vías metabólicas, etc. El objetivo del presente trabajo con ovocitos de sapo Bufo arenarum es estudiar los mecanismos de acción de sustancias xenobióticas a nivel bioquímico y molecular, en relación con la capacidad de éstas gametas de ser fertilizadas y avanzar a través de los estadios tempranos del desarrollo embrionario. Se ha trabajado con diversos biomarcadores que permitieron reconocer los mecanismos de toxicidad de compuestos organoclorados y más recientemente de algunos metales pesados. Se demostró que la activación de una fosfolipasa C específica por el Dieldrin provoca depleción de mediadores lipídicos y como consecuencia inhibición de la fertilización (desarrollo hasta 4-8 células). Actualmente se estudian los efectos del Zn sobre la fertilización y su probable relación con alteraciones en la actividad de enzimas de la vía de las pentosas, directamente ligada a los mecanismos de detoxificación en presencia de stress oxidativo. El estudio de los mecanismos de citotoxicidad de diversas sustancias resulta de gran utilidad cuando se lleva a cabo en gametas, porque permite también evaluar los efectos de dichas sustancias sobre la reproducción de la especie. Los ovocitos de anfibios son además células de gran interés, por su utilidad en el estudio de la citotoxicidad de una amplia variedad de contaminantes hídricos (AU)

Acumulación de Zn en ovocitos de sapo Bufo arenarum: efecto sobre el metabolismo de carbohidratos / Accumulation of Zn in oocytes of Bufo arenarum frog: effect about the carboydrates metabolism

La exposición de hembras de sapo Buffo arerarum a aguas del río Reconquista (Provincia de Buenos Aires, Argentina) conduce a la acumulación de zinc (Zn) en sus ovarios. Las hembras tratadas de esta forma son capaces de ovular cuando son inyectadas con macerado de hipófisis, al igual que hembras controles. Los ovocitos obtenidos de hembras tratadas son capaces de fertilizar y desarrollar normalmente hasta el estadío de gástrula, mientras que presentan un 27 por ciento de inhibición del desarrollo embrionario a partir del estadío denominado respuesta muscular. In vivo la microinyección de Zn en ovocitos controles simultáneamente con [U-14C] Glucosa evidenció disminución en la síntesis de glucógeno y en la oxidación de la glucosa a través de la vía de las pentosas. La actividad de la enzima glucosa-6-fosfato deshidrogenasa en ausencia de agregado de Zn fue de 371,8 x 10 -3 U/ml.min. La enzima fue inhibida en vitro por el metal en forma dependiente de la concentración. Utilizando una concentración igual a 1,53mM de Zn, similar a la incorporada en ovario y a la microinyectada en los ensayos con glucosa radiactiva, se alcanzó un 62 por ciento de la máxima inhibición correspondiente a 3mM de Zn (248,7x10-3U/ml.min). Estos resultados concuerdan con un efecto inhibitorio del Zn sobre el desarrollo embrionario, mediado probablemente por una deficiente producción de NADPH, ribosa-5 fosfato y ATP en los ovocitos.

Is aging related to cell signalling alterations in amphibian oocytes?

O armazenamento de oócitos de sapo Bufo arenarum diminuiu sua habilidade para serem fertilizados "in vitro". A estimulaçäo com carbachol em "oócitos jovens" mostrou uma hidrólisis persistente de fosfatidilinositol 4,5 bifosfato (PIP2) näo foram hidrolizados até um nível significativo. Estes resultados e os menores níveis de 32P-fosfoinositideos achados nos "oócitos envelhecidos" num tempo zero de estimulaçäo, condordam com uma diminuiçäo nas atividades das quinases de fosfoinositideos e da fosfolipase C, como consequência de uma hidrólisis näo específica de fosfoinositidos que ocorreria durante o armazenamento